8th European A A V ConferenceAssociation of Avian Veterinarians 6th Scientific E C A M S Meeting European College of Avian Medicine and Surgery In collaboration with AFVAC Genac Association Française des Vétérinaires pour Animaux de Compagnie Groupe d’Etude des Nouveaux Animaux de Compagnie Arles, France, April 24-30, 2005 AAV AFVAC ISBN : 2-9508885-9-3 AFVAC - 40 rue de Berri - 75008 PARIS - FRANCE Phone : + 33 (0) 1 53 83 91 60 - Fax : + 33 (0) 1 53 83 91 69 - Email :
[email protected] Association of Avian Veterinarians European committee (EAAV) www.eaav.org, www.aav.org European College of Avian Medicine and Surgery (ECAMS) www.ecams-online.org Association Française des Vétérinaires pour Animaux de Compagnie (AFVAC) www.afvac.com Groupe d’Etude des Nouveaux Animaux de Compagnie (GENAC) www.afvac.com 8th European A A V Conference 6th Scientific E C A M S Meeting Arles, April 24-30, 2005 In collaboration with AFVAC Genac EAAV CONFERENCE CHAIRMAN Jean-Marie Péricard, DVM, Certified in Avian Diseases, Certified in Epidemiology - Clinique vétérinaire, Sigean, France EAAV SCIENTIFIC COMMITTEE CHAIRMAN Jaime Samour, MVZ, PhD, Dip ECAMS Fahad bin Sultan Falcon Center, Riyadh - Kingdom of Saudi Arabia ECAMS SCIENTIFIC COMMITTEE CHAIRMAN Nico J. Schoemaker, DVM, PhD, Diplomate ECAMS, Diplomate ABVP (Avian) Division of Avian and Exotic Animal Practice, Utrecht University, The Netherlands AAV EUROPEAN CONFERENCE 2005 COMMITTEES 26-30 APRIL 2005 ARLES PROVENCE FRANCE ORGANISING COMMITTEE Jean-Marie PERICARD (France), Chairman of the organizing committee Lorenzo CROSTA (Italy, Spain), EAAV President Peter SANDMEIER (Switzerland), Chairman, 2007 EAAV Conference Helga GERLACH (Germany), Treasurer Niel FORBES (United Kingdom), EAAV-ECAMS delegate SCIENTIFIC COMMITTEE Jaime SAMOUR (Saudi Arabia), Chairman of the scientific committee Jean-Pierre ANDRE (France) Tom BAILEY (Dubai, UAE) John CHITTY (United Kingdom) Gerry DORRESTEIN (Netherlands) Helga GERLACH (Germany) Nigel HARCOURT-BROWN (United Kingdom) Jean-Michel HATT (Switzerland) Rüdiger KORBEL (Germany) (and website coordinator) Suzan OROSZ (USA) Jean-Marie PERICARD (France) LOCAL ORGANISING COMMITTEE Jean-Marie PERICARD Michel BELLANGEON Franck RIVAL (President of GENAC) Didier BOUSSARIE Emmanuel RISI Guillaume LELOCH Gérard GELLY Jean-François QUINTON Sylvie DUFOUR (Administration director AFVAC) Didier-Noël CARLOTTI (President of AFVAC) ECAMS ORGANISING COMMITTEE M. Krautwald-Junghanns (Germany) Nico J. Schoemaker (Netherlands) EDITORS EAAV Tom BAILEY John CHITTY Nigel HARCOURT-BROWN Jaime SAMOUR AUDITORS Peter COUTTEEL (Belgium) Nigel HARCOURT-BROWN (United Kingdom) SPONSORING We greatly appreciate the support by our exhibitors and sponsors : For this conference : BAYER HEALTHCARE VETOQUINOL & ELLMAN International Inc. HARRISON’S BIRD FOODS HEALTH AND HYGIENE JANSSEN SANTE ANIMALE KARL STORZ LYON ELECTRONIC CO. INC. MEDLAB MELET SCHLOESING Laboratoires OPTOMED – EVITEC OXBOW PET PRODUCTS SCANELIS SCIL ANIMAL CARE COMPANY VERSELE-LAGA For the pre-conference scientific tours : LPO (Ligue pour la protection des oiseaux) BIRDLIFE INTERNATIONAL RESERVE NATURELLE DE CRAU STATION BIOLOGIQUE DE LA TOUR DU VALAT For the practical laboratories : ALCYON BIRD-BITS CENTRAVET ELLMAN International Inc. ESCULAPE GENIA GER KARL STORZ MEDICAL SOLUTION NIKON SCIL ANIMAL CARE COMPANY VETOQUINOL 3M Welcome to Arles 2005 ! The French Companion Animal Veterinary Association (AFVAC) and its exotics study group (GENAC) are happy and proud to contribute to “Arles 2005”, a joint event with AAV and ECAMS. The GENAC is a very active group of our Association. They took up the challenge of such a meeting with enthusiasm and competence. Bravo! Many thanks also to AAV and its Board for their effective collaboration with our national group. Bird medicine and surgery has become a well recognized discipline. This is due to the incommensurable involvement of dedicated veterinarians. This has justified the creation and development of ECAMS, a species-oriented European College of Veterinary Specialists, third partner of the adventure. Thank you as well… When a field expands, congresses are the indispensable stages of its development. This one will be remembered, I am sure, because of the outstanding and sophisticated programme set up by the organizers. Plenary Sessions, Master Classes and Practical Laboratories are the corner-stones of a well-balanced scientific meeting like this one. I am also convinced that the remarkable environment of Provence will contribute to the success of this gathering of highly motivated colleagues. Please take at least a few hours to enjoy the place… I thank deeply all the organizing team and particularly Jean-Marie Péricard who spared no effort and time to make this event possible and, I am already convinced, successful. I wish you all a pleasant and fruitful meeting. Didier-Noël Carlotti, President of AFVAC A conference « mirror » of our activity It wasn’t the aim, but it is the result : This conference reflects the role played by the avian veterinarian within our profession and society. The main role of the avian veterinarian is to treat and prevent the diseases of privately owned pet birds, pet shop birds, breeding centre birds, wild birds as well as those birds found in zoological parks, rehabilitation centres and reintroduction programs. By doing it, this veterinarian participates in the protection of the sanitary state of the poultry industry, and in the prevention and epidemiological survey of the zoonosis. But the veterinarian has also, in these different situations, a role of zootechnician, nutritionist and behaviourist. As a matter of fact, all these aspects are dealt with during this conference ! The participation of more than 160 colleagues from 21 countries in the 2 pre conference scientific tours, 71 oral presentations, 2 round table discussions, 6 master classes, 26 posters and 9 practical labs, displays, beyond our diversity, the similarity of our care to improve a scientific field in rapid progression. I wish everybody to make use of this conference, but also to enjoy Provence and the exceptional birds living in these top places of biodiversity in Europe that are the Camargue, the Crau and the Alpilles. Jean-Marie Péricard, Chairman of the organising committee Table of Contents Scientific meeting of the European College of Avian Medicine and Surgery S.I.C.O. Santos & J.T. Lumeij. “Concealed” avian sexual dichromatism and multiple angle reflectance spectrometry ......................................... J.-M. Hatt & J.T. Lumeij. Evidence-based avian medicine – How to ask questions and find answers ............................................................. R. Korbel & K. Sturm. Review on lightsources for birdhousing under artificial light circumstances .................................................................. U. Benheim & I. Davidson. Molecular Identification and Prevalence of Psittacine Circovirus in Israel ............................................................ M. Hochleitner et al. Fungal cultures in feather probes of psittacine birds ...................................................................................................... S.E. Orosz. The senses of birds : their unique qualities ...................... J. van Engelen, I. Akkerdaas & N.J. Schoemaker. A study into the analgesic efficacy of Buprenorphine and Butorphanol in pigeons (Columba livia domestica) .................................................................... E.S. Story, D. Carboni & T.N. Tully. Establishing normal tear production values in hispaniolan Amazon parrots (Amazona ventralis) .................... J. Samour. Haemopathological responses in falcons to Newcastle disease ................................................................................................. M.-E. Krautwald, G. Stelzer & V. Schmidt. Food - intoxication in nestling psittacines ............................................................................... I. Westerhof, N.J. Schoemaker & P.Y. Barthez. Spiral Computed Tomography in Respiratory diseased Birds ....................................... T.A. Bailey & O. Combreau. Domestication and Disease : Challenges for Houbara Bustard Captive Breeding Projects in the Middle East ...... B.W. Ritchie et al. Epizootiology of Proventricular Dilatation Disease in Breeding Cockatiels .......................................................................... 3 5 7 9 11 13 19 21 23 25 27 29 31 Main European Association of Avian Veterinarians Conference J. Chitty. Veterinary aspects of the great bustard (Otis tarda) Reintroduction project in the UK. ......................................................... M. Barrows, M. Hartley, J.M. Pittman. Diseases and management of captive crested screamers (Chauna torquata) ....................................... G. Cousquer. Clinical approach to the downer swan ........................... U. Höfle, J.M. Blanco, R. Valboa, D. Villanua, C. Gortazar. Trichomonas gallinae in free-living birds of prey, passerines birds and woodpigeons (Columba palumbus) ......................................................................... E. Jourdain, M. Gauthier-Clerc, D. Bicout, Y. Kayser, P. Sabatier. A study of free-ranging wild birds to better understand their role in the circulation of west nile virus in Camargue, Southern France .................. O. Lambert, H. Pouliquen, B. Philippe, N. de la Cotte, M. L’Hostis. Wild bird exposure to anticoagulant rodenticides in Loire-atlantique (France), a 2 year study on waterbirds and raptors (2002-2003) .......... G. Riggs Mycobacterial infection in waterfowl collections : a conservation perspective ............................................................................................. M.G. Muller, A. George, A.T. Mannil. Haematological values of gyr hybrid falcons ......................................................................................... J. Samour, J.L. Naldo. Causes of morbidity and mortality in captive falcons in Saudi Arabia ......................................................................... T.A. Bailey, C. Silvanose, E. Pesci, A. Di Somma. Histoplasma sp. incidental finding, underdiagnosed, or occasional opportunistic pathogen of falcons in the Middle East ? ............................................................ U. Wernery, F.A. Hassan, T.A. Bailey, B. Johnston, J. Kinne. Mycobacteriosis in hunting falcons in the Middle East ................. S. Le Dréan-Quenec’hdu, L. Marion, M. Popoff Epidemiology of avium botulism in France : example of Grand Lieu Lake ....................... A. Y Gancz, I.K Barker, L. Robbin, A. Dibernardo, K. McKeever, B. Hunter. A West Nile virus outbreak in North American owls ................. 37 41 47 56 60 66 70 77 85 92 99 106 112 ... A............. Gelli. M.T Lumeij............ I... R... K................ G. F......... Olsen.. N......... H.. Petroselli... Central nervous symptoms caused by thiamine deficiency in juvenile goshawks (Accipiter gentilis) .... Ferrari... Dorrestein..... Chavernac... Forbes..........A Hafez. M..... Adenovirus infection in raptors .J............... F......... S.... Schuffenecker......... M Bedin. Auge. Zientara... Lierz...... Romagnoli..... M............... Circovirus infection in a canary breeding flock ..................... Monks.... D. Franceschini. Henning............ Biomolecular study of a progression of haemocromatosis in hill mynahs (Gracula religiosa) . .......... V...... G.......... G.J Schoemaker. Exocrine pancreatic insufficiency in racing pigeons (Columba livia domestica) ...J Henning..M..... D......... and treatment of iron storage disease using the European starling as a model G.... E...... Pesaro. D.. Schmidt. Zsivanovits.. H.. Risi. S... N...... external skeletal fixator and tension bands : a pilot study in domestic pigeons (Columba livia domestica) .... O. A.... J.. R... M...... Amann... J. J.............. D. Bargalló............... but several dietary factors and bird-specific defense mechanisms contribute to the development of the disease ......... S. G.... F. Surveillance of West Nile virus in the avifauna of Southern France .........M. I... Serum biochemical and electrophoretic patterns in the Eurasian buzzard (Buteo buteo) – Reference values .....J. young pigeon disease....... P.... Rodriguez-Quiros................ C... Gonzalez....... Ordonneau.. prevention.... Westerhof...... Lierz... Management of the femorotibial luxation by coaptation splinting.. Lopez Peña. O..... A comprehensive study on a disease complex associated with pigeon circovirus infection......... Bernardini......... M............ Balanca........... intramedullary pins... Mortamais..... Rossi. J....... J... M-E. Phalen. Freire.. Ramis........ Two cases of femoral head luxations on cormorants (Phalacrocorax carbo) treated with a modified Meij-Hazewinkel-Nap technique. S............ Diagnosis...... Benito de la Vibora.. Peccati....... H.. Hafez. Perpiñán.. Carnarius................................... Villaverde. P....... Endoscopic guided multiple entry surgery in birds .. D. M...... V...... Ballester... Grífols............ N....M Visschers..... 120 130 136 141 148 151 159 166 171 179 184 190 196 ......... J. Gauthier.... Hars............. Zeller..... Muñoz Guzon....... Pradel....... D. P...... Ceccherelli.... D..... Werquin Hepatic haemosiderosis in birds : not only total dietary iron level.... Russell........... Krautwald-Junghanns. ..... H. M............. .........M......................... Scope.. Use of a dental composite to correct beak deviation in Psittacine species .. M..... Bretagne.. A.......... Lublin. G.. 203 207 211 217 222 230 233 238 243 249 254 257 260 ............... G. side effects and efficiency .... Pharmacokinetics... S.. bioavailability and compatibility of doxycycline in birds after oral administration .. Pees. Mitchell................. E............ Haefeli..... Pethel...........H... S.. W................. Reitz. K...........S......... M............. Burhenne. K..... Krautwald-Junghanns........ Guillot.. The use of enalapril in birds...E............ Korbel. D...............C..... Deville....... H... C..A..... M-E... Pharmacokinetics and pharmacodynamics of the new antifungal agent voriconazole in birds ....... Desmarchelier............ S.......... Evaluation of the serological test Platelia® Aspergillus for the diagnosis of aspergillosis ......... Wilson..... M-E.. E.. M. N. Risi......... Zachariah.......... Monitoring of end-expired partial pressures of carbon dioxide in anaesthetised birds of prey ..... M.........H... J............ L.... Fitzgerald......... Perpiñán........... A................. Clinical assessment on the use of fluconazole per os in 24 African grey parrots (Psittacus Erithacus) : acceptance..... Krautwald-Junghanns.........M....... S.. V......... G........ Schmidt. Krautwald-Junghanns..... Tully......... Oral furazolidone and chloramphenicol for treatment of gastro-intestinal bacterial infections .. Pharmacokinetics and use of meloxicam in psittacine birds ............... Evaluation of various tissues for diagnosis of psittacine beak and feather disease (PBFD) ..... N.............. Péricard. Stevens... B.... Weilacher.. M...........A. Hess...... M. Latimer...... F......... Buerkle....Figaroa....... The effects of isoflurane anaesthesia on gastrointestinal transit time ...R.. Fernández-Bellon... Edery....... Hernandez-Divers. O..... C............... R.. A.. M-E.......... U. Rondenay. Indications. clinical experiences. Lair.. pharmacokinetics and possible side effects ...T.... J.. Grund...... Reliability of crop biopsy as a diagnostic tool for proventricular dilatation disease in psittacines birds ... Ungemach. T......................... Ramis.T...................... Le Loch... A.....N.... Bendheim. Budsberg.... J.. Crosta.. Kuhring............... Köhler......... Diaz ..... M.....M......... Lennox... Myenteric ganglioneuritis in four non-psittacine birds .. López........ Y...... ....... Chitty. Chaste-Duvernoy...... Mechani................ Roman...K. G....... The nutritional implications of producing the «optimal egg» in captive birds ........ R.... S..... Roman........... H......... Stanford.......................................... Rival.. Liesegang..D................... Forbes.................... C... A.. 267 274 277 283 290 298 301 307 310 314 318 321 327 333 340 348 .. Hatt.................. Reavill........................... Zsivanovits........ Crosta.. A..... A review of avian endoventricular mycosis submitted during 1998-2004 .... S........ Monks... Smith..... Plasma electrophoresis reference ranges in various bird species .................... R..... Forbes........ Pesaro. Monks.............. R.. Tarantino.. Early detection of an acute inflammatory condition by plasma protein electrophoresis and haematology in peafowls ....... Y...... Scoccianti..... Diagnoses of rotavirus in pet birds vs Poultry ..... H............... N... Bumbarov.........M.................................... Gerlach.......... Ceccherelli.......... D... Müller........ D...................... Timossi. E. Digital scanning ophthalmoscopy in birds ...... D... T....... Stockdale......... L.... Phylogenetic analysis of the hexon loop 1 region of adenovirus isolated from psittacine birds indicates the existence of a new psittacine adenovirus ... B..............D... J-M... Dorrestein.. Y.... N................ Chaste-Duvernoy.. S.. Assessment of disc fusion tests to evaluate the effects of antibacterial nebulisation agents against bacteria isolated from the avian trachea ...... Bert.A... S...... Consequence of severe dyspnoea for organ function ..... Biosecurity of the avian hospital facility by design.............. C.....Lublin.............. L......... The first Italian outbreak of polyomavirus infection in macaws ........... Ordonneau....... D..................................... P. M........T..... Cloacal disease and disorders in the avian patient ...... Imaging techniques in avian obstetrics ... Rossi.. Korbel......... G. D... V............ I... Zsivanovits..... J............. F.... Ritzman....A..... D...... Raue. Quantitative computed tomography for the assessment of bone density in budgerigars : a pilot study .. protocols and procedures ......... Ordonneau.. P...................... Fischer................. The effect of UV-B radiation on calcium metabolism in grey parrots .. Fisk... Auricular diseases in birds . ........... Hooimeijer..... P... Le Maho............ Kirby....... beak and feather disease and avian polyomavirosis 354 359 365 374 388 409 425 434 447 451 452 453 ... Macwhirter..... M................. J-M............ S........... fluid therapy and CPCR for the avian patient ... E... Real-Time PCR for diagnosis of three common infectious diseases in caged birds : Chlamydiophylosis..... Gauthier-Clerc..... Parrots don’t bite children ....................................... Lafon....... D.. C............................. Infestation by ticks Ixodes uriae in penguin colonies .. M...... Avian clinical anatomy from an evolutionary perspective ................. Inman.. C................. Concurrent Master Classes sessions P. Cray. Page. The avian respiratory system : normal and pathological aspects ...................M....... Feldman............. Le Bohec..... Le Maho....... S....... Orosz.... L...... J-L.......... L...... S. Gauthier-Clerc.............. P...... Effects of flipper bands on free-living penguins . Dorrestein.............. Chavez.. J-P... Keller. M.. Influence of a seed diet versus a formulated diet on bodyweight and intestinal flora in budgerigar (Melopsittacus undulatus) over a six month period ......... J............................ Y.... Shock................ A master class of the urinary system : from anatomy to treatment options . W.. Diehl.....M.................. Passerine and softbill medecine and surgery . Y.... Imaging non-invasive diagnostic techniques of the urogenital tract .............. Zwart.......... C... C..... Gendner..... S.......... M-E. Pingret..... B......... C.. Pees............ Lichtenberger..... Echols................ Krautwald-Junghanns............. Lichtenberger. Le Bohec.............. Boucraut-Baralon.. Mull... G. Orcutt...... Stumpp.... Posters European Association of Avian Veterinarians Conference Fischer.... Response to fluid resuscitation subsequent to acute blood loss in mallard ducks (Anas platyrhynchos) ........ DeBehnke... R................. C....... Hatt . M........ J.... Di Somma...... Computed tomographic anatomy of the pigeon (Columba livia) ...... S........ A..... Kammerer............... P.. M. Bailey... Mckinney. I........ 456 459 461 464 467 470 473 476 479 482 484 488 491 495 ........ S.... Wenery... A. Hartley................ Le Dréan-Quenec’hdu................. Endo –parasites of the Asian houbara bustard (Chlamydotis undulata macqueenii) . support cushion to facilitate wound healing in bumblefoot .. M... Cause of mortality of wild birds collected in a wildlife center in France : example of Nantes Wildlife Center .. Lambert.......... Determination of the possible exposure of raptors and waterbirds to five anticoagulant rodenticides in Loire Atlantique (France 44) ........ Albaric O... M.. Blanco................ F........ Vanadium content of oiled birds : is it a good marker for oil exposure ? ..M................... Risi E...... Zinc toxicosis in a wattled crane (Bugeranus carunculatus) .... Remple................ van Bree........ Pasmans........... Van Caelenberg. T............ Tena..... C........ Villaverde................... E. Costiou... C............ Mucocele in a lesser sulphur-crested cockatoo (Cacatua sulphurea) . Lambert......... Health control of various capercaillies (Tetrao urogallus) captured in the Principality of Andorra .... Pouliquen.........S.. Risi.. Silvanose..... Gielen.. Treatment of crop fistulas due to severe trichomoniasis in two booted eagles (Hieraaetus pennatus) ... H............................................ C......... N. J........... J-P. H. Barrows... H. L’Hostis.... U. de la Cotte............ Schuster.. André................ R. U.... Successful treatment of a cervical rhabdomyosarcoma on an Indian Fantail pigeon (Colomba livia) by surgical resection .......... Pouliquen.................................... Höfle. J.......................... Congenital abnormalities in a wild young Tawny owl (Strix aluco) .......... Chitty... M........A. Degorce-Rubiales.... O..M. Le Dréan-Quenec’hdu............. J. Pittman.. A......... F... Abadie J...... J............. M... O.... Kinne..... A new method for the cytological sampling of feather pulp ........ Antifungal susceptibility testing of fungi isolated from the respiratory tract of falcons .... E... flexible............... Martel......... NGuyen F... Risi. Nurse............. P... Use of a composite silicone dental impression material to create a form-fitting....... Larhantec ........ Galvez Aguilera.............. H................ PruknerRadovcic.......... Lennox.. T.............. 498 501 504 507 509 512 515 518 Practical Laboratories European Association of Avian Veterinarians Conference List of the practical laboratories ....... A.................. Gaillet..................J.......................................... M. Roberts........... Fisher..... Chronic polychondritis in an African grey parrot (Psittacus erithacus) .. E........................ Mazija................... Acosta Guevara.. Ixodes frontalis as a cause of avian disease in the UK ....... Grozdanic............... Stanford.................. S... Post-intubation stenosis in psittaccine birds ........... J... M........ Forbes... 523 ...........................A............ Latimer...... Ritzman........................... N...... Wilson................... H....... C.............. Clinical approach to ophthalmologic disorders in birds ............ Soto Piñeiro........... R...... Horvatek.. Gottstein...... The role of avian medicine in a parrot conservation project of Amazona leucocephala in Cuba : preliminary results .......... Bert.... Hernandez-Divers........... Terrrier.......K.... Rival. C. E... Barrat......... K. L..... N.......M... Klebsiella oxytoca infection in monk parakeet ................... Diseases found in French birds according to SAGIR database ............... Gonzalez Navarrete.. M.... Northrup........... F.. Z.. D..... Tomassone....... Monks.......A... J-R........... X.. Radiation tolerance doses of the cutaneous and mucosal tissues in psittacine birds using cobalt-60 teletherapy ..... I..... M-E.. . . Scientific meeting of the European College of Avian Medecine and Surgery ABSTRACTS . . Santos. were assessed with a spectrometer using different illumination (45º and 90º) and observation angles (30º.O. Resulting reflectance data were summarized in several parameters (brightness.C. colour intensity.Lumeij. All avian species studied thus far by spectrometry. and were analysed by logistic regression. 3 . 75º.Division of Avian and Exotic Animal Medicine. PhD ABSTRACT Numerous avian species have plumage that reflects both in the human visible range and in the ultraviolet (UV) part of the spectrum. 45º. Different body regions. the influence of angle geometry in the colour assessment of non-iridescent feathers has been proved to be significant as well.It has been shown that birds employ their ability to detect UV light. including the Long-tailed finch previously classified as monomorphic by Langmore & Bennett (1999). Considering that angle geometry does have an influence on spectrometric results and that birds exhibit different positions towards the opposite sex during mating displays. One hundred percent correct sex prediction could be achieved by combining different colour parameters from different body regions and angle geometries. The results of this study will not only have impact on avian morphology studies but also all subsequent theories in different fields of biology such as evolution and behaviour. and Magpie (Pica pica). All studied species. Long-tailed finch (Poephila acuticauda). Faculty of Veterinary Medicine. apart from foraging and navigation. either from the researcher or the studied subject. in behavioural interactions and mate-choice. have been proven to be sexually dichromatic when the UV part of the spectrum was considered. 90º. The Long-tailed finch has been classified by single angle spectrometry as monochromatic. both in the UV and in the total range. Java finch (Padda oryzivora). proved to be sexually dichromatic. with the exception of the Long-tailed finch (Poephila acuticauda) (LANGMORE and BENNETT. DVM. Most studies have only used one angle geometry (illumination/observation) to assess noniridescent plumage colours. Utrecht University “CONCEALED” AVIAN SEXUAL DICHROMATISM AND MULTIPLE ANGLE REFLECTANCE SPECTROMETRY S. from the Blue fronted Amazon parrot (Amazona aestiva). since their colour changes with the observation/illumination angles. In iridescent feathers. 60º. even when the UV part of the spectrum was considered. actively involved in sexual displays. Department of Clinical Sciences of Companion Animals. contrast and UV chroma). we hypothesize that all diurnal avian species are sexually dimorphic and that this trait can be visualized by using multiple angle spectrometry. 1999). hue.T. J. 135º).I. Reflectance spectrometry is considered an accurate and objective way to assess plumage colour and its results are independent of the colour perception system. However. more than one angle geometry has been used. AUTHORS ADRESS S. Santos Division of Avian and Exotic Animal Medicine Department of Clinical Sciences of Companion Animals University of Utrecht Yalelaan 8 3584 CM Utrecht / The Netherlands e-mail: s.I. Proc R Soc Lond Ser B . LANGMORE N E and BENNETT A T D.Biol Sci 1999.O.REFERENCES 1.i.nl 4 .c.o. Strategic concealment of sexual identity in an estrildid finch.santos@vet. 266: 543 – 550.C.uu. Comparison and Outcome) introduced. age. A special challenge in applying EBM is the translation of a clinical problem as encountered in practice. PhD ABSTRACT This paper presents current definitions of evidence-based medicine (EBM) and proposes to adopt for veterinary medicine the following modified definition by Cockroft & Holmes (2003): Evidence-based veterinary medicine is the use of current best evidence in making clinical decisions. Previously given examples will be used to demonstrate retrieval of evidence. Department of Small Animals Vetsuisse Faculty University of Zurich. into an answerable clinical question.nih. Lumeij. which is a spectrum of potential sources with the source most likely to provide the best evidence at the top. Example: How does PCR compare to ELISA in the diagnosis of Chlamydophila psittaci? Outcome: The outcome that is desired may have to be included in the question. species.-M.org). using these databases will be presented. DVM. Of special interest is the Medline database (Pubmed. Example: Lead intoxication in free ranging vultures. Patient: By categorising e. The process of developing a well-formed clinical question is explained and the key elements (Patient or Problem.cabi-publishing. 5 . Example: Is there an enhancement of ventriculotomy healing in Amazon parrots after inclusion of a collagen patch during surgery? Subsequently sources of evidence will be discussed and the hierarchy of evidence is introduced. Example: Will an iron level of <150 ppm in mynah diets significantly reduce the incidence of iron storage disease? Comparison: Searching for best evidence often implies a choice between two interventions or an intervention and no treatment at all. in relation to aspects such as time-frame. welfare. Hatt.Division of Zoo Animals and Exotic Pets.pubmedcentral.T. Prof Dr med vet. Switzerland EVIDENCE-BASED AVIAN MEDICINE – HOW TO ASK QUESTIONS AND FIND ANSWERS J. Intervention: Definition of the diagnostic or therapeutic intervention and the exposure or prognostic factor that the patient has been or will be subjected to. J. Intervention.g. cost-benefit. www. management (captive or free). a search in databases will be more focused. the approach to answer a specific clinical question and to retrieve general information quickly. www.gov) and the Veterinary Science Database (formerly VetCD. Blackwell Publishing. Oxford. 2003 AUTHORS ADDRESS Division of Zoo Animals and Exotic Pets Department of Small Animals Vetsuisse Faculty University of Zurich Winterthurerstrasse 260 8057 Zurich Switzerland Jean-Michel.unizh.Further reading COCKROFT P and HOLMES
[email protected] 6 . Handbook of evidence-based veterinary medicine. Dr med vet habil. K. The table on the following page gives a quick orientation of light sources available on the market. which are kept under artificial light sources. Korbel. with light that fulfils their needs regarding to vision aspects. Ludwig-Maximilians-University Munich. Sonnenstrasse. Furthermore birds are able to resolve motion pictures at a higher flicker frequency (approximately 180 frames / sec) rather than man (15 – 80 frames / sec).pet birds as well as poultry./ tetrachromatic vision including ultraviolet perception. Oberschleissheim. as detection of sex via UV-reflection of feathers. incandescent light as well as discharge lamps. assessment of food sources and prey patches. pigeons and raptors . Prof Dr med vet.de 7 . which to a certain extent meet avian requirements: AUTHORS ADRESS Ruediger Korbel. LED (light emitting diodes)-light sources. feeding of chicks are based on UV-vision and thus are of importance not only regarding to animal welfare aspects. These circumstances have to be considered when keeping birds . Prof Dr med vet.Klinik für Vögel.under artificial light circumstances. Dr med vet ABSTRACT Unlike man most of the diurnal birds are capable of penta. energy saving light sources. Cert Vet Ophthalmol Klinik für Vögel Ludwig-Maximilians-University Munich Sonnenstrasse 18 D – 85764 Oberschleissheim / Germany e-mail: korbel@lmu. Following a review on avian vision capacities known so far the proposed paper will give a technical orientated review of commercially available light sources including fluorescent lamps. Germany REVIEW ON LIGHT SOURCES FOR BIRD HOUSING UNDER ARTIFICIAL LIGHT CIRCUMSTANCES R. Sturm. These data will be compared and discussed regarding their suitability providing birds. ballast 2. ballast 3% UVB none none (absorbs radiation below 390nm) 80% in infrared yes full-spectrum nearUV to infrared yes according to according to filter ballast natural daylight according to practically none (filter) spectrum ballast narrow-band spectrum 380-700nm. ECG: operates at 30-50 megahertz frequencies ³ DC: operates under direct current 8 .Light sources according to their suitability for birds Light source Brandname Incandescent Conventional Spectral range Flickerfree* UVA portion 400 -1100nm.120 Hz. peak at 410nm yes (DC³) yes (DC) none none * Flicker frequency over 180 Hz ¹ no UVA according to Thrush 2000 ² CCG: operates at twice the mains supply current: 100 .4% UVB according to 10% UVA. max at 950nm yes none not described¹ ESU Birdlife Brightlight Spot full-spectrum Incandescent Lamp® Fluorescent + CF Conventional True-light-Solux® Vita-lite (Duro-Test) Activa (Sylvania/Osram) Arcadia Birdlamp ESU Avian Birdlamp Halogen Conventional DECOSTAR IRC cold-light reflector lamp (Osram) HID Conventional SoLux® LED Conventional Golden Dragon (Osram) 3-bandspectrum full-spectrum full-spectrum full-5-bandspectrum full-spectrum full-spectrum yes according to none ballast² according to described ballast according to described ballast according to described ballast according to 12% UVA. Jerusalem. The virus was detected in both laboratories with a similar efficacy. including cockatoos and other psittacines. Most positive cases were detected in clinically-affected birds. Hebrew University.7% in the following 2 years. I. Kimron Veterinary Institute. The birds tested for PBFD by PCR in the years 2001-2003 showed the following results: All Psittacines Cockatoos Grey Parrots Other Psittacines 13.7 % positive n=417 n=126 n=142 n=149 During the years 2001-2003 the infectivity rate increased from 6. feather loss and necrosis of the upper beak (BENDHEIM and HOCHLEITHNER. most birds suspected for PBFD infection have shown typical clinical signs. The diagnosis was confirmed by histopathology and electron microscopy.Koret School of Veterinary Medicine. 1991). In several cases PBFDV sequences were detected in apparently healthy birds. imported from Singapore. South Africa (MDS). Since then. Bendheim.3% in the year 2001 to 18. DVM. This was done from blood samples taken from the brachial vein. Division of Avian and Fish Diseases. Davidson. 9 . During the last 4 years PBFD is being monitored routinely in 4 breeding farms and some Psittacines collections in order to detect apparently healthy PBFD carriers and are assessed by the bird types and years. while afterwards it was performed also in the Kimron Veterinary Institute for research purposes. where the Ypelaar PCR (YPELAAR et al.9% and 19. The diagnosis was confirmed by PCR in different laboratories. Israel MOLECULAR IDENTIFICATION AND PREVALENCE OF PSITTACINE CIRCOVIRUS IN ISRAEL U.9 % positive 28.9 % positive 6. 1999) was applied and validated by sequencing. Bet Dagan. Until December 2003 the PBFD infectivity was assessed in 417 birds by PCR performed only by the Molecular Diagnostic Services in Westville.0 % positive 4. PhD ABSTRACT Psittacine Beak and Feather Disease (PBFD) was first detected in Israel in two Sulphur-Crested Cockatoos. who showed typical clinical signs: feather deformation. we analyzed feathers that were shed in the proximity of clinicallyaffected birds and were able detect viral-sequences in several cases. Vet Microbiol 1999. I.. and HOCHLEITHNER M. AUTHORS ADDRESS U. M. 68: 141 – 8. WILCOX. G. A universal polymerase chain reaction for the detection of psittacine beak and feather disease virus.R.R. RAIDAL.E.. Bendheim. Unpublished case 1991. YPELAAR. and S. Box 196 Shawe Zion 25227 Israel 10 . BASSAMI.O. 2. DVM P. REFERENCES 1.To broaden the molecular detection capability and the environmental monitoring of circoviruses. BENDHEIM U. N. Dr med vet. brought to the clinic for other reasons (African Grey Parrots (n=48). DVM ABSTRACT Microbiological testing in cases of feather picking in psittacine birds is often recommended as standard procedure. Ziegler. Leidinger.000 Cockatoos Amazon Macaws Budgerigar Total 15 2 14 5 82 1 8 3 0 18 0 2 1 1 13 19 30 10 14 194 0. In this study feather probes (feathers plugged 1cm lateral of christa sternalis both sides) of feather picking and non-feather picking birds of different psittacine species were collected and incubated on sabourauds glucose 2% agar. Amazon parrots (n=32). Amazon parrots (n=10).000 0. cockatoos (n=19). macaws (n=11) and budgerigars (n=15)). C. Dr med vet. Positive fungal cultures of Aspergillus sp.000 0. Hochleithner. E. The study included birds with the history and signs of feather picking (African grey parrots (n=52).Tierklinik Strebersdorf. Austria FUNGAL CULTURES IN FEATHER PROBES OF PSITTACINE BIRDS M.236 0. are considered to be controversal.000 Tab. Harrison. Differences between frequency distributions of culture results of feather-picking and non feather picking birds were tested for significance with Fischer Exact test (Table 1). Vienna. With the exception of Amazon parrots the results demonstrate a significant increase in prevalence of Aspergillus sp growth (within 72 hours) on feather probes from feather picking birds compared with those from non picking birds. G. cockatoos (n=16). 11 .000 0. Hochleithner. macaws (n=17) and budgerigars (n=5)) as well as birds with no history or signs of any feather or skin problem. Species Feather picking Non feather picking P value of difference Fisher Exact test Aspergillus pos neg pos neg African Grey Parrot 46 6 9 39 0. 1 Aspergillus sp culture results from feather samples collected from feather plucking psittacines and psittacines without feather disorders. VWR International). First of all positive test results can be observed in non-feather picking birds. Staining was performed by a cytological standard staining (Hemacolor. Significant higher numbers of positive cultures were found using the secretion of the uropygeal gland which seems to be in contrast to literature were this gland secretion should prevent fungal growth on feathers in birds. the fact that feather picking birds show a higher frequency of positive cultures might be secondary to poor feather structure caused by feather picking. A few slides were additionally stained by Gram-stain (VWR International).at 12 . Hochleitner. In a second study feathers of 40 African grey parrots were collected and incubated on sabourauds glucose 2% agar which had been prepared with the content of the uropygeal gland of each bird as well as without this agar modification. making the feathers vulnerable to secondary infections. Dr med vet Tierklinik Strebersdorf Muhlweg 5 Vienna. Finally the secretion of the uropygeal gland was examined histologically. Furthermore.The clinical significance of positive culture results is not clear. AUTHORS ADRESS M. AT 1210 / Austria e-mail: hochleithner@aon. determine ripeness largely by taste and smell rather than by sight. 2000). These factors need to be taken into account clinically as humans. The cross-sectional diameter of the optic nerves of birds are larger than the diameter of their cervical spinal cord (KING and McCLELLAND. Raptors have even larger numbers of neurons per optic nerve as their acuity surpasses that of other living beings (FOX and WESTENDORF.Perrysburg Animal Care Perrysburg. It is in the retina where light energy is converted to electrical impulses by “bleaching” of the photoreceptors. BREAZILE and KVENZEL. identify birds within their own species as individuals. appears visually as sexually monomorphic. for example. 1999) that. 1969. vision is by far the most important.1984. The shape of the globe of a particular species of birds is the result of its ecological requirements (GÜNTÜRKÜN. PhD This paper will review the special senses of birds with particular emphasis on aspects with clinical applications. These components of the globe or optic media are very transparent and are able to transmit light down into the near ultraviolet spectrum (at least 310 nm) (EMMERTON. Orosz. to us humans. This large collection of myelinated axons is from the ganglion cell layer of the retina. The eyes of birds comprise a considerable volume of the head and are very large in relation to brain size. and to determine the sex within a species (KORBEL and GROPP. and PAULE. 1961. The visual impression of objects and colors are important to birds. Ohio USA THE SENSES OF BIRDS : THEIR UNIQUE QUALITIES Susan E. DVM. 1942). humans are highly visual primates but have only 40% of the numbers of retinal axons per optic nerve when compared to pigeons or chicks (BINGGELI. 1978). Part of this results from the shape 13 . PORTMAN and STINGELIN. This ability to see into the near ultraviolet allows them to discern ripeness of food items. thereby allowing the optic image to be spread out over a larger retinal surface. Birds are unique and wonderful creatures and their senses are an integral part of their makeup. Birds are exquisitely visual animals. RAGER and RAGER. 1976). anterior chamber. which is the largest of any land vertebrate and twice that of a human (WALLS. The basic components of the eye are similar to mammals except for the scleral ossicles to help maintain the shape of the globe. In comparison. and the vitreous body before is reaches the retina. 1993). Light has to pass through the cornea. Acuity is maximized by increasing the anterior focal length of the eye. Another technique that achieves the same goal is represented in the eyes of raptors. The eye of the ostrich has an axial length of 50 mm. Their tubular-shaped globes result in high visual acuity. lens. 1980). Of all the senses of birds. they evolved neuronal mechanisms to master equivalent cognitive demands (DIEKAMP et al. It sits dorsally over the midbrain. GÜNTÜRKÜN. 2000). However. We need to think about this level of complexity when developing toys and enrichment items as they are very color oriented. 2000). We are just beginning to unravel the vast complexities of their visual system and its relationship to learning and memory. Sensitive hearing and vocal communication are important in the behavioral repertoire of birds. This area may also be involved in pecking and food selection. they appear to use their neostriatum caudolaterale. 1992. It was supposed that working memory--the ability to temporarily store and manipulate currently relevant information-. Clinically. Memory of visual images and working memory are also observed in birds. They are able to rank optic patterns by using transitive inference logic (von FERSEN et al. birds are able to handle working memory without a laminated neocortex. these daylight raptors have reduced resolution at dusk (RAYMOND. this may make it difficult for the bird in a low-light hospital cage to locate its food. 1988) as well as “same” versus “different” (WRIGHT. 1985). The normal range for hearing often does not exceed 10 kHz but it does have a restriction to an upper limit of 20kHz (SCHWARTZKOPFF. The lowly pigeon has not been considered the smartest bird but they have been found to understand visual concepts of “animals” (ROBERTS and MAZMANIAN.600 caches of stores containing 33. 1972).was the consequence of a prefrontal cortex and a neocortex containing grey matter. It is also involved in enhancing contrast under dim light conditions (HELLMANN et al. there are some birds that are able to hear infrasound (NECKER. 1988). Birds have the most highly evolved auditory system of non-mammalian species. and they know humans as individuals as well. Consequently. we must bear in mind that they have very active visual brains that need stimulating. they are also great discriminators of thousands of images. As with some mammalian species. as occurs in mammals. 14 . 1995). When we approach their needs. that does not mean that the avian ear is inferior (NECKER. From the enrichment perspective. 2000). but it may also allow the bird to be calmer than at higher light levels. However. 2002).000 seeds (Vander WALL and BALDA. This is a relay station for visual information. Studies have shown that birds such as the Clark’s nutcracker have visual memory for over 6. 1973). Information from the retina is sent to the area that is represented as the superior collicus in mammals. Instead. this non-pooled system requires high light intensity to function adequately. While the mammalian ear is considered to be more specialized than birds. 1977). We know clinically that parrots know the difference between individual dogs. Data demonstrates that even though they do not possess the same anatomic components as mammals. as they love to call them by name. This area is so large in birds that it is described as the optic lobe. This reflex pathway is thought to relay information concerning moving objects (ROGERS and MILES. like the elephant.and the remainder is from the high ganglion cell to receptor hookup ratio that increases visual resolution. 2000). and motion in the membrane sets up a wave motion in the membranous labyrinth of the inner ear. as is their ability to discriminate 2 sounds of different intensity (KLUMP and BAUR. These birds use this sense in dark caves for orientation (NECKER. it appears on the initial contact. Owls have the ability to discriminate changes in the location of sound sources that are as small as 3 degrees apart and can aim their heads within 2 degrees of the source. Echolocation occurs in only two families of birds. Chemesthesis is the perception of chemically induced pain. Two sounds separated by a gap are recognized as separate if the gap exceeds 2–10 msec (WILKINSON and HOSE. 1980). It has also been determined 15 . the mammalian basal ganglia and its connections with the thalamus and cortex are important for motor control and cognitive functions. This has important implications from a lab animal perspective and a humane one as well.Hearing involves complex interactions between the sense organ. They have a spatial map in their midbrains that is much larger than their behavioral perception. and the processing of sound in the central nervous system. It seems that birds rarely avoid mammalian irritants even though the trigeminal nerve is responsive to the chemical stimuli (MASON et al. This suggests that birds. have a comparable ability to learn and should not be relegated to second class citizen status in the animal world. 2001). MASON and OTIS. vocalization. the columella. The sensory input for this chemical avoidance sense in birds appears to be the components of the trigeminal nerve (CN V). The wave motion is converted to mechanical energy by the discharge of sensory receptors located on the papilla. and there is no evidence that aversion is altered by GI feedback. The tympanic membrane is attached to its ossicle. Time resolution of hearing in birds is also similar to humans (DOOLING. The electrical energy and its discharges are transmitted to the CNS by the eighth cranial nerve (VIII). Airborne sound waves enter through the external acoustic meatus and cause the tympanic membrane to vibrate. The ability to discriminate frequency differences in birds is similar to that of humans (KUHN et al. 1990). These chemicals are aversive on a purely chemical basis. 1975). 1992). 2003). Hearing represents the sensory component to the motor response. It appears that the avian brain has a closed loop for vocal learning that is similar to that of mammals (LUO et al. 1989. This aversive quality is unlearned. Further. which are represented by the oilbird and by swiftlets. the ear. In the CNS. For example. like mammals. Sensitivity to chemical irritants is adaptive so that they avoid actual physical damage by avoiding noxious stimuli. birds do not seem to associate aversion of the stimulus with other chemosensory cues. the Steatornidae. Studies of vocalization and vocal learning are providing further information about the avian brain and its processing. 1995). further processing occurs and is interpreted by higher centers. the hair cells. Many aromatic chemicals are aversive to birds. For example. The chemical senses of bird include their ability to taste and their ability to smell. a typical neuron has a spatial receptive field that spans 40 degrees – many times wider than the behavioral threshold (BALA et al. 1990). suggesting that conditioned flavor avoidance learning does not occur (CLARK. indicating that issues involving pain and its management should be addressed in a manner similar to other companion animals. We should also consider their visual perception (eg. The avian taste bud does not open directly onto the epithelial surface. While in the past. the thermoreceptor. Taste buds are distributed throughout the oropharynx. The mechanoreceptors. recent data shows that these preconceived notions are not true. oral medications may continue to be refused despite frequent attempts to habituate a bird to their taste. Taste preferences.that birds do not habituate to the stimulus so that the avoidance persists without reinforcement (CLARK and SHAH. 1990). and the nociceptors (pain) serve different but important functions. Although most of the chemical sense of birds is toward avoidance of a noxious compound. STROMBERG and JOHNSEN. Birds have a greater capacity visually than humans and a similar capacity with hearing. both by the texture and pressure and by the heat generated. It is important for the clinician to note that the pain pathways appear to be similar in birds when compared to mammals (OROSZ. 1990). need to be considered both in their husbandry and medical treatment. While we humans may enjoy something or project that a bird may like it. Many species including parrots. As a result. their taste preferences are not well documented. Cockatiels are more sensitive to organic acids than to inorganic acids (MATSON et al. for example. Birds without a nasal salt gland tend to refuse foods with a concentration that is hypertonic to their body fluids (BARTOLOMEW and MacMILLAN. suggesting that saliva would be important to the transference of the chemical sense to a mechanical nerve ending associated with the taste bud. Birds have unique anatomic features including their lissencephalic brain. 1996). Finches in the family of Carduelidae have a great preference for salt (MASON and ESPAILLAT. 16 . or perhaps. budgerigars and nectar feeders have a preference for sweetness using natural sugars mixed with drinking water (KARE and ROGERS. will require us to be remain creative about medicating birds. Birds have relatively few taste buds when compared to other vertebrates. The somatosensory system allows for the animal to perceive its body surface with the external environment. The information received by the exteroreceptive cutaneous receptors is taken into the spinal cord to be sent up to the level of the telencephalon for interpretation. Bird brains and their sensory systems are not that simple! Birds’ cognitive and learning abilities. ability or inability to discern food in ambient light and calming effect of low levels of light) when treating or convalescing birds. The Herbst corpuscles allow ducks billing in the water to discern the texture of the items in the murky water. 1985). 1994). that you are dremeling its beak. This data suggest to the clinician that food preferences are individualized and that they tend to avoid inorganic acids while sweetness may help to hide drugs and other medications. taste aversion. our taste preferences may serve the bird poorly. that feature suggested bird brains were not endowed with the mental capacity of mammals and humans. They also provide information to a macaw. 1960). Birds may be tolerant of sour but this tolerance is species dependant. They are not taste or smell oriented but their somatosensory system is similar neuroanatomically to humans. 1976. often in close association with salivary gland openings (BERKHOUDT. along with their pain perception. 2001). BREAZILE JE. Systema nervosum centrale. Tests and refinements of a general structure-activity model for avian repellents. GÜNTÜRKÜN O. Naturwissenschaften 1980. Cytochrome oxidase activity revelas percolations of the pigeon’s ectostriatum. DOOLING R. San Diego. 13. CLARK L. 545 – 59. PERKEL DJ. DING L. Prediction of auditory spatial acuity from neural images on the owl’s auditory space map. 137: 1 – 18. ESPAILLAT JE. BALA AD. 22(4): RC210. BINGGELI RL. Nuttall Ornithological Club No. 19. KVENZEL WJ. MUTH I. 424: 771 – 4. MACMILLAN RE.) Structure and function of avian taste receptors. MA. 19: 1382 – 7. Sensory physiology: Vision. LUO M. 463 – 96. POPPER AN (eds): The Evolutionary Biology of Hearing. BAUR A. In: STURKIE PD (ed) Avian Physiology. Ultraviolet perception in birds. Their Structure and Function. 77: 545 – 7. J Neurosci 2001. NY: Springer-Verlag 1992. Bailliere Tindall 1984: 237 – 314. Differences in taste preference between redwinged blackbirds and European starlings. In: WHITTOW GC (ed): Sturkie’s Avian Physiology. FOX R. 271: 432 – 40. 20. Nature 2003. WALDMANN C. 9. Proc Annu Conf Assoc Avian Vet. 7. TAKAHASHI TT. Hearing in birds. (1985. 3. Naturwissenschaften 1990. 10. KARE MR. GÜNTÜRKÜN O. FAY RR. 17 . 8. LEPPELSACK HJ. Science 1976. 67: 102. KORBEL RT. Academic Press 1985. 11. In: LEVY AS. MCLELLAND (eds): Form and Function in Birds III. 77-81. 20: 321 – 39. Physiol Zool 1960. The pigeon retina: quantitative aspects of the optic nerve and ganglion cell layer. DIEKAMP B. J Exp Zool 1995. 17. SCHWARTZKOPFF J. 21(17): 6836 – 45. 171. WESTENDORF DH. CLARK L.REFERENCES 1. EMMERTON J. New York. SPITZER MW. KLUMP GM. 192: 263 – 5. Spectral transmission of the ocular media of the pigeon (Columbia livia). Wilson Bull 1990. LEHMKUHLE SW. 14. KALT T. GÜNTÜRKÜN O. ROGERS JG. 4. 5. 18. SHAH PS. MCCLELLAND J. Intensity discrimination in the European starling (Sturnus vulgaris). 12. 5th ed. In: WEBSTER DB. SCHLECHT P. 23. Sense organs: taste. NY. 6. Birds. KUHN A. Modulation of avian responsiveness to chemical irritants: effects of prostaglandin E1 and analgesics. J Chem Ecol 1994. In: BAUMEL JJ (ed): Handbook of Avian Anatomy: Nomina Anatomica Avium. The water requirements of mourning doves and their use of sea water and NaCl solutions. J Neurosci 2002. 2nd ed. 15. MASON JR. (1999). Measurement of frequency discrimination in the starling (Sturnus vulgaris) by conditioning of heart rate. 2. PAULE WJ. GROPP U. Cambridge. BERKHOUDT H. 102: 292 – 9. 16. New York. BARTOLOMEW GA. Bath. Falcon visual acuity. Working memory neurons in pigeons. Neuroreport 1995. Invest Opthalmol Visual Sci 1980. 33. Berlin. 493 – 554. CA: Academic Press 2000 HELLMANN B. J Comp Neurol 1969. Springer Verlag 1976 KING AS. 6: 881 – 5. 1993. United Kingdom. An avian basal ganglia pathway essential for vocal learning forms a closed topographic loop. SCHWEMER J. 5th ed. The avian ear and hearing. KING JR. CLARK L. Vander WALL SB. Anthranilate repellency to starlings: chemical correlates and sensory perception. PhD. WALLS GL. Transitive inference formation in pigeons. SCHWARTZKOPFF J. RIVERA JJ. 48: 147 – 56. Centrifugal control of the avian retina. 33: 65 – 78. V. optical and anatomical investigation. MATSON KD. 20: 1 – 13. 36. 23. and sour taste stimuli in cockatiels (Nymphicus hollandicus). AUTHORS ADRESS S. 25. Aquila audax: A behavioral. MI 1942. COOK RG. Thresholds for sweet. Principles of avian clinical neuroanatomy. New York. 258: 320 – 1. 53: 55 – 64. KLASING KC. 26. Orosz. The central nervous system. vol II. 30. ROGERS JJ.” Cranbook Institute of Science. Hummingbird sweetness preferences: taste or viscosity. A quantitative electronic microscopic study of the generation and degeneration of retinal ganglion cells in the chicken. monkeys. STROMBERG MR. NECKER R. 1996. 37. 17: 334 – 41. ROBERTS WA. WRIGHT AA. DELIUS JED. WILKINSON R. RAYMOND L. ADAMS MA. Systems-matching by degeneration. Bloomfield Hills. Wilson Bull 1990. J Wildl Manage 1989. Concept learning by pigeons: matching-to-sample with trial-unique video picture stimuli. CA. 31. 14: 247 – 60. Ecol Mon 1977. Effects of lesions of the isthmo-optic nucleus on visual behaviour. Mechanoreception. Zoo Biol 2001. J Exp Pyschol Anim Behav Process 1988. MASON JR. Ohio USA e-mail: DrSusanOrosz@aol. New York. OROSZ SE. Avian Exotic Pet Med. Academic Press 1961: 1 – 36. In: WHITTOW GC (ed): Sturkie’s Avian Physiology. 34. JOHNSEN PB. Concept learning at different levels of abstration by pigeons. Coadaptation of the Clark’s nutcracker and the pinyon pine for efficient seed harvest and dispersal. Anim Learn Behav 1988.21. 102: 292 – 9. 417 – 77. 29. Academic Press 1973. Vision Res 1985. HOSE PE. salt. and people. Condor 1990. 25: 1477 – 91. SANDS SF. DVM. 32. I. RAGER G. 33. 22. 27. PORTMAN A. 16: 436 – 44. DELIUS JD. “The Vertebrate Eye and Its Adaptive Radiations. Nature 1975. Exp Brain Res 1978. OTIS DL. Semin. PARKES KC (eds) Avian Biology. ABVP-certified in Avian Practice Perrysburg Animal Care Perrysburg. MAZMANIAN DS. RAGER U. von FERSEN L. 47: 89 – 111. Brain Res 1972. 32: 606 – 12. MILES FA. WYNNE CDL. In: FARNER DS. San Diego. Spatial visual acuity of the eagle. BALDA RP. MASON JR. NY. 35. 24. STINGELIN W. STADDON JER. Academic Press 2000. MILLAM JR. Dipl. 28. In: MARSHAL AJ (ed): Biology and Comparative Physiology of Birds. 5: 127 – 39.com 18 . Time resolution of acoustic signals by birds. Aversiveness of six potential irritants on consumption by red-winged blackbirds (Agelaius phoeniceus) and European starlings (Sturnus vulgaris). J Exp Psychol Anim Behav Process 1992. DVM. Schoemaker. DVM. 1994). butorphanol (2 mg/kg) or saline (control) with a fixed volume of 0. and was found to be more effective in providing analgesia than buprenorphine in conscious African Grey Parrots (PAUL-MURPHY et al. At least one week was scheduled between each study to allow the birds to fully recover. Premedication comprised either buprenorphine (0. Department of Clinical Sciences of Companion Animals. PhD ABSTRACT In the last decade opioid drugs have been studied for their efficacy in providing analgesia in avian species. there was no significant difference in heart frequency in any of the pigeons during the noxious stimuli. The Netherlands A STUDY INTO THE ANALGESIC EFFICACY OF BUPRENORPHINE AND BUTORPHANOL IN PIGEONS (Columba livia domestica) J. Anaesthesia was maintained at an end-tidal concentration of 1. the latter being determined randomly according to the Latin Square.J.5 mg/kg).7 ml IM. In this study. Each pigeon was studied 3 times. 1984. The study was performed in 9 pigeons. University of Utrecht. since in mammals there is evidence that cardiovascular and pain regulatory pathways are functionally linked (RADICH and MAIXNER. after which the pigeons were intubated and ventilated with intermittent possitive pressure (IPPV). DVM. In Germany. change of heart rate was used as the measure for analgesia. I. 19 . During the experiment heart rate. van Engelen. Akkerdaas. Fourty-five minutes after premedication. Reactions to the stimuli were also registered. every time with a different form of premedication. N. Since butorphanol was not readily available in The Netherlands the authors wanted to compare the analgesic efficacy between butorphanol and buprenorphine in pigeons anaesthetized with isoflurane. 2000) as well. Although the heart rate in pigeons receiving butorphanol was significantly lower compared to those receiving saline or buprenorphine. The investigator was unaware of the premedication the pigeon had received prior to measurements. body temperature and end-tidal concentrations of CO2. 3 times with three minute intervals. The total volume was divided over both pectoral muscles. Most studies have been performed in the United States of America were it was found that butorphanol has an isoflurane-sparing effect (CURRO et al. 1999). Faculty of Veterinary Medicine.Division of Avian and Exotic Animal Medicine. O2 and isoflurane were monitored. a noxious stimulus (toe pinch) was administered for a period of 10 seconds.8 % isoflurane. ZAMIR and MAIXNER. however. it was found that buprenorphine in a dose of 0. One hour after premedication. 1986). Differences between the heart frequencies in the three groups were compared by means of an ANOVA for repeated measures.5 mg/kg had an analgetic effect in racing pigeons (GAGGERMEIER et al. anaesthesia was induced by mask with isoflurane 4% in 100% oxygen (1l/min). The role of sinoaortic and cardiopulmonary baroreceptor reflex arcs in nociceptive and stress-induced analgesia. PhD. domestica).). 1789. The relationship between cardiovascular and pain regulatory systems. 3. 60: 1218 – 21.J. Determination of the ED50 of isoflurane and evaluation of the isoflurane-sparing effect of butorphanol in cockatoos (Cacatua spp. München 2000: 75 – 6.J. GAGGERMEIER B. RADICH A and MAIXNER W.nl 20 . 23: 429 – 33. ZAMIR M. REFERENCES 1. HENKE J. New York Academy of Science 1986. PAUL-MURPHY J. New York Academy of Science 1984. SCHATZMANN U. AUTHORS ADRESS N. BRUNSON DB. 5. 2. Untersuchungen zur Schmerzlinderung mit buprenorphin bei Haustauben (Columba livia Gmel. Vet Surg 1994.uu. PAUL-MURPHY J. BRUNSON DB. MAIXNER W. 4. ABVP-certified in Avian Practice Division of Avian and Exotic Animal Medicine Department of Clinical Sciences of Companion Animals University of Utrecht Yalelaan 8 3584 CM Utrecht / The Netherlands e-mail: N. MILETIC V. Deutsche Vet Gesellsch. var. DVM.. 371 – 384. Analgesic effect of butorphanol and buprenorphine in conscious African grey parrots (Psittacus erithacus erithacus and Psittacus erithacus timneh). Schoemaker@vet. 385 – 401. as assessed by an increased heart rate response to a standard noxious stimulation. Am J Vet Res 1999. Schoemaker. ERHARDT W. In: KELLY DD (ed): Stress-induced Analgesia. Dipl. KORBEL R.We therefore conclude that no analgesic effect could be measured in pigeons receiving either buprenorphine or butorphanol (at the trial dose rates). In: KELLY DD (ed): Stress-Induced Analgesia. CURRO TG. Mississauga. 1994. Based on prior research reports. Louisiana. USA) was used. For statistical analysis of tear production measurements paired t-test were used to compare PRTT and STT values prior to and following topical anesthesia. 2) PRTT measurements of tear production will be easier to perform in birds. Corneal anesthesia was accomplished using topical proparacaine after the initial STT and PRTT and prior to a second STT and second PRTT. To perform the PRTT. 5mm filter paper strips were used (Schirmer Tear Test Strips. the Schirmer Tear Test (STT) and the Phenol Red Thread Test (PRTT). Tear absorption and a resulting red color change on the thread was measured using Jameson calipers. DVM. and to compare PRTT and STT values before and after topical anesthesia in Amazon parrots. Ontario. The PRTT has been investigated in dogs and cats (BROWN et al. MS. but will have a wider standard deviation than the STT. To compare 21 . 1997). MS ABSTRACT There are currently 2 methods used to measure tear production in animals. to determine the mean and normal range of PRTT values in normal Amazon Parrots prior to topical anesthesia (PRTT1) and after application of topical anesthesia (PRTT2). To perform the STT. Canada) and maintained behind the ventral eyelid margin for 1 minute. After all tests were performed a sodium fluorescein dye was used to identify any corneal ulceration or trauma.. 1996. No investigations have been performed to confirm the viability of the PRTT in avian species. Alcon Canada. Debra Carboni. DVM. DVM. Story. San Mateo. two primary hypotheses were proposed for this investigation: 1) STT1 (without anesthesia) and STT2 (with topical corneal anesthesia) will not be significantly different from previous reports. Thomas N. Phenol Red Thread Test – Zone Quick ® (Menicon America.Louisiana State University – School of Veterinary Medicine Baton Rouge. placing a 3mm bend in the thread then inserting into the lateral canthus for 15 seconds. The purpose of this study was to determine if the PRTT is a feasible measurement of tear production in birds. California. Tully. Jr. United States ESTABLISHING NORMAL TEAR PRODUCTION VALUES IN HISPANOILAN AMAZON PARROTS (Amazona ventralis) Eric S. Twenty – six (26) Hispanoilan Amazon parrots maintained at the LSU – School of Veterinary Medicine in a controlled environment allowed for the first study of this type using a statistically significant population of a similar psittacine species with a known life and medical history. The PRTT is faster and causes less reflexive tearing than the STT due to the significantly less irritating contact with the cornea in species where it has been evaluated. KORBEL and LEITENSTORFER. There have been 2 reports of values obtained when performing STT in birds (WILLIAMS. 1996). KORBEL R. Florida: Wingers 1994. RESULTS PRTT1 PRTT2 STT1 STT2 15.edu 22 . DVM LSU – School of Veterinary Medicine Dept. BRIGHTMAN AH.3. The phenol red thread tear test in healthy cats. WILLIAMS D. 51: 171 – 5. REFERENCES 1.lsu. 3. DAVIDSON HJ. BROWN MH. Avian ophthalmology. BROWN MH. 7: 249 – 52. 4. In: RITCHIE B.2. DAVIDSON HJ. GALLAND JC.6 mm 8. LEITENSTORFER D.the individual birds from day to day the proc GLM test for repeated measures was used.05 was considered significant for our statistical results. We also determined that the 5mm commercial STT test strips could be used to measure tear production in Amazon parrots with results almost identical to previous studies using modified STT test strips. Lake Worth. 673 – 94. Vet Comp Ophth 1996. HARRISON LR (eds): Avian Medicine: Principles and Application.2. 6: 274 – 7. Clinical estimation of lacrimal function in various bird species using a modified Schirmer tear test. P< 0. BRIGHTMAN AH. Israel J Vet Med 1996. Pearsons correlations between PRTT & STT were also determined during this investigation.97) but did significantly effect the STT values (p < 0. Baton Rouge. Corneal anesthesia did not significantly effect PRTT values (p = 0.5 mm (8-22) (7-23) (3-12) (1-12) [9-22] [8-23] [3-13] [0-10] PRTT1 – phenol red thread test without topical corneal anesthesia PRTT2 – phenol red thread test with topical corneal anesthesia STT1 – Shirmer tear test without topical corneal anesthesia STT2 – Shirmer tear test with topical corneal anesthesia Statistical results of this study indicate the PRTT is a feasible measurement of tear production in the Amazon parrot and can be used to identify Amazon parrots with low tear production. Based on our findings the PRTT test is a feasible option to measure tear production in Amazon parrots and most likely other avian species.5 +/.3 mm 15. 2. HARRISON GJ.1 +/.3.3 mm 5.0 +/. AUTHORS ADDRESS Eric Storey. 70803 LA / United States e-mail: storeye@vetmed. The phenol red thread tear test in dogs.5 +/.000). Vet Comp Ophth 1997. GALLAND JC. VCS Skip Bertman Dr. for further studies. PhD ABSTRACT Newcastle disease (ND) is an infectious and highly contagious disease which commonly affects falcons in Saudi Arabia and the Middle East as a whole. UK. lung. Newcastle disease virus was isolated in all 34 falcons. regurgitation and vomiting. metallic-green coloured urates. Clinical signs associated with the viscerotrophic form vary also in type and severity but commonly include initially reduce to total absence of appetite. The disease is thought to be highly prevalent subsequent to the extensive feeding of pigeons throughout the region. gastroscopy and radiology examinations. head tremors and tics or sudden death. The pathological appearance of the ventriculus of affected individuals could be observed during gastroscopy and at radiology in 65% of the cases. No other gross pathological finding could be made. ND is produced by Paramyxovirus 1 (PMV 1). regurgitation and vomiting. hyperesthesia. progressive paralysis of the legs. clinical signs. Clinical symptoms varied accordingly to the clinical presentation. amaurosis.8-11. heart. Riyadh.43 ± 1. Haematology analysis was carried out in 34 falcons suspected of undergoing Newcastle disease based on the anamnesis. All 34 falcons died 3 to 8 days after the clinical diagnosis was made. reduced to total absence of appetite. A selection of tissues including brain. clonic spasm. Falcons affected with the viscerotrophic form showed a slightly enlarged liver and a grossly distended ventriculus. constant distress vocalisation.39 x 109/l) (normal 3. shredding and flicking of food. Samour MVZ. paresia or paralysis of the nictitant membrane. the vomiting of partially digested blood or sudden death. Fahad bin Sultan Falcon Center. metallic-green coloured urates leading to dysphagia and salivation due to tongue paresia.Falcon Specialist Hospital and Research Institute. Post-mortem examination of cases affected with the neurotrophic form revealed only a slightly enlarged liver. mucoid diarrhoea. ataxia. kidney and spleen were submitted for virus isolation. The walls of the ventriculus were thickened and petechial haemorrhages were observed in most cases in the isthmus and in the ventriculus. Laboratory results from 30 falcons revealed moderate leucocytosis (12. The clinical signs of falcons affected with the neurotrophic form varied in type and severity but could include initially. leading to severe depression. Isolates were sent to the World Reference Laboratory in Weybridge. All falcons have been fed on freshly killed or live pigeons in the past 7 to 14days. Kingdom of Saudi Arabia HAEMOPATHOLOGICAL RESPONSES IN FALCONS TO NEWCASLE DISEASE J. shredding and flicking of food.5 x 109/l) with absolute 23 . liver. 5. ALEXANDER DJ. CAMPBELL TW. Newcastle disease. 5(3): 21 – 24. Ames: Iowa State University Press 1998.33 ± 1. Norwell: Kluwer Academic Publishers 1988. The haematology findings highlighted the importance of expressing the results of the differential white cell count in both absolute and percentage values. Riyadh 11322. 253 . REFERENCES 1.heterophilia (84. Lake Worth. WERNERY U.45 x 109/l) with moderate heteropenia (50 ± 4. RITCHIE BW. 3.O. Kingdom of Saudi Arabia e-mail: falcon@shabakah. MVZ. London: Mosby 2000. 264 . Newcastle disease in free-living and pet birds. In most cases the heterophils showed signs of toxicity (grades 3 and 4) including basophilic cytoplasm.05 %) and severe monocytosis (31.246. PhD Falcon Specialist Hospital and Research Institute Fahad bin Sultan Falcon center.19. 197 . AUTHOR ADDRESS Jaime Samour. 3 . In: Samour J (ed) Avian Medicine.com 24 . P. State Vet J 1995. 2. absolute heterophilia was only detected when the percentage of heterophils in the blood films was taken into consideration.8 ± 0. In: ALEXANDER DJ (ed): Newcastle Disease.3 %. normal 0-4 %). loss of granulation and loss of lobulation. Fl: Wingers Publishing Inc 1995.33 ± 8. 2nd edition. In all cases examined the absolute heterophil count was within established normal ranges. Function and Control. normal 60-68%). Avian Hematology and Cytology. The toxic changes observed in the heterophils were considered typical of the acute phase of the ND infection. Box 55. Avian Viruses.311. 4. Newcastle disease. KALETA EF and BALDAUF C. However.266. Four falcons showed moderate to severe leucopenia (1.1 %. -E. The food as well as the birds´ livers and kidneys were examined for their vitamin and mineral-content. After having used their psittacine diet 26 nestling birds of different species died within 5 months. 25 . It was noted that the food (compared to the recommendations by the National Research Council and the AAV´s Expert group) contained an increased Vit A. A time consuming extensive and costly examination followed. Stelzer. Some birds which showed a 5 fold increased content of Vitamin A in the liver (levels about 7200 I. Consequently there was a strong suspicion of food related intoxication. which corresponded partially to an increased content of these vitamins in the liver /kidneys of the dead birds. in which different specialized veterinary institutions as well as 5 different veterinarians were involved. crop stasis and liver alterations. Krautwald-Junghanns. The investigations contained a gross necropsy of all dead psittacine nestlings. A variety of changes were recorded including (visceral?) gout. parasitological./ g liver) for example did not exhibit any typical pathological findings. The pathological findings were different in the various birds without any apparent common theme. these findings could not be directly connected to hypervitaminosis A/ D3 or E. A – level in the liver.INTOXICATION IN NESTLING PSITTACINES M. G. cytological. On the other hand some birds fed with the same food showed a decreased Vit.content. Germany FOOD . V. Additionally the food was examined for mycotoxins and for the composition of amino-acids. In addition histological. microbiological and virological examinations were performed. After changing the food no further mortality was noticed. In the following noteworthy case. However.Clinic for Birds and Reptiles University Leipzig. a psittacine breeder took legal proceedings against a food company.E. it is difficult to prove this suspicion without a thorough veterinary investigation at quite an early stage. Schmidt ABSTRACT Food intoxication is often suspected in a case of abnormal mortality in bird flocks. Prof Dr Med vet. which resulted in the death within a few days of the positive group (fed with the food under trial). Further to this food trials with budgerigars were undertaken. Vit D3 and Vit E . However. as well as selenium . This corresponded again to partially increased contents of the minerals mentioned in liver and kidneys of the nestling birds. .de 26 . Krautwald-Junghanns Clinic for Birds and Reptiles Department of Small Animals University Leipzig An den Tierkliniken 17 04103 Leipzig Germany e-mail: krautwald@vmf. Dr.uni-leipzig. The decision of the court will also be presented and discussed. copper. AUTHORS ADRESS Prof. zinc.In relation to mineral content. M.-E. the content of the different vitamins and minerals in the organs and their interaction between each other as well as the pathological findings are discussed taking into account of the different species and age of the birds. The relationship between these findings in the food.levels were elevated when compared to the published recommendations. Diagnosis of the exact cause of the respiratory distress is difficult. In the literature computed tomography has been described as a non-invasive diagnostic tool (KRAUTWALD-JUNGHANNS et al. DVM. i. In this study birds were scanned during inhalation anaesthesia with isoflurane to facilitate precise positioning of the birds. Spiral CT requires less examination time under anaesthesia than conventional radiography or conventional computed tomography. Faculty of Veterinary Medicine. PhD.Westerhof DVM. This study describes spiral CT in 20 birds with respiratory problems. CT findings are described and compared with conventional radiographic findings. KRAUTWALD-JUNGHANNS et al. The CT images provided superior information of the respiratory tract compared to the radiographs.Division of Avian and Exotic Animal Medicine. Schoemaker. DVM. P. On the other hand. helical or in volume acquisition. The Netherlands SPIRAL COMPUTED TOMOGRAPHY IN RESPIRATORY DISEASED BIRDS I. University of Utrecht. KRAUTWALD-JUNGHANNS et al. CT is an imaging technique which has. 1997). with a total scanning time of less than 1 minute per bird. In these studies sagital cross sections were used as the standard view. 1998[3]. The number of scan slides used in the birds were at least 250. In 10 birds the findings are also compared with post mortem findings. PhD ABSTRACT Respiratory problems occur frequently in birds..e. ROMAGNANO and KRAUTWALD-JUNGHANNS. 1998 [2]. Department of Clinical Sciences of Companion Animals. PhD.J. spiral CT generates many detailed CT images that requires more interpretation time and experience from the interpreter of the CT images. CT scans of respiratory diseased birds are compared with those of healthy birds. Spiral CT. Barthez. Spiral CT enables scanning of a complete organ during the time of maximal enhancement without artefacts caused by respiratory movements (SOUCEK et al. 1998[1]. thus far not been described in birds.Y. N. small lesions may be missed or misinterpreted. were included in the standard scanning protocol. The individual results will be discussed during the presentation. 27 . 1990). Conventional radiography is usually performed in order to identify abnormalities in the respiratory tract. using radiography. Birds were fixed in sternal recumbency. starting at the beak and ending at the tail. However. Transverse total body cross sections of the whole bird from cranial to caudal. It is concluded that spiral CT provides detailed information about alterations of the respiratory tract in birds. SCHUMACHER F. 3. PhD Division of Avian and Exotic Animal Medicine Department of Clinical Sciences of Companion Animals University of Utrecht Yalelaan 8 3584 CM Utrecht / The Netherlands e-mail: I. Westerhof.uu. KRAUTWALD-JUNGHANNS ME. 1: Examination of healthy parrots. Examination of the lower respiratory tract of Psittacines and Amazoniae varieties by means of reconstructive transmission computed tomography. 17 – 21.nl 28 . Clinical applications. Examination of the lower respiratory tract of Psittacines and Amazoniae varieties by means of reconstructed computer x ray tomography. Röntgenpraxis 1990. SOUCEK M. 65(1): 17 – 22. KRAUTWALD-JUNGHANNS ME. 1998. Spiral CT: a new volume scanning technique. SOHN HG. KRAUTWALD-JUNGHANNS ME. II. DVM. KALENDER W. Westerhof@vet. AUTHORS ADRESS I. Respiratory Radiology and Imaging. VALERIUS KP. SCHUMACHER F. 2: Examination of parrots with respiratory symptoms. Tierarztl Prax Ausg K Kleintiere Heimtiere. 5. SOHN HG. 1997. Tierarztl Prax Ausg K Kleintiere Heimtiere 1998. CT-assisted versus silicone rubber cast morphometry of the lower respiratory tract in healthy amazons (genus Amazona) and grey parrots (genus Psittacus). VOCK P. DUNCKER HR. 1998. 43: 365 – 75. 4. Proc Avian Spec Adv Prog & Small Mamm and Reptile Med and Surg.REFERENCES 1. 26(2): 139 – 49. ROMAGNANO A. 26(1): 61 – 70. Res Vet Sci. 2. DAEPP M. SOHN HG. KRAUTWALD-JUNGHANNS ME. endangering captive-breeding populations. Confiscated ‘smuggled’ wild-caught birds that have come through an official rehabilitation programme and are placed in captivity. 2) considering health when founder stock are selected. Health issues vary according to the source of the bird. exposing the birds to new environments and diseases. we conducted health surveys on wild and rehabilitated bustards including: Free-living bustards . PO Box 23919. United Arab Emirates DOMESTICATION AND DISEASE : CHALLENGES FOR HOUBARA BUSTARD CAPTIVE BREEDING PROJECTS IN THE MIDDLE EAST T. Conservationists are concerned that novel diseases or genetic conditions may be transferred between captive and wild populations by these movements. houbara bustard farming. The larger projects have started to address these issues by initiating veterinary programmes that include. By managing bustards under intensive captive breeding conditions man has started the process of domestication. Bailey. In the context of this information the health challenges posed by domestication are assessed and veterinary recommendations for what may become a new regional industry. 1) investigating the health of wild populations. rehabilitated and captive houbara in the region. 3) managing the health of stock during the breeding programme and 4) health screening during the release stages. Bustards maintained in captivity in the Middle East are derived from four sources: 1) 2) 3) 4) Wild caught birds smuggled directly into collections in the region. Dubai. Rehabilitated bustards . Consequently. Officially sanctioned projects collecting eggs from wild populations.A. Pakistan and the UAE) of live and dead (hunted) free-living bustards were examined from 1993 to 2001.the health status of confiscated bustards in a survey between 1992 and 1999. As a consequence captive bustards are vulnerable to a wide range of disorders. are presented. Exchanges of breeding stock between the larger captive breeding projects. PhD.samples collected from three populations (China. Houbara captive breeding programmes involve managed movements of animals between captive and free-living populations. MRCVS. 29 . released animals and free-living populations. there has been a surge of interest in the propagation of bustards in captivity. Combreau ABSTRACT In recent years. O. This presentation reviews health surveys conducted on free-living.Dubai Falcon Hospital. The research requirements of the young houbara bustard ‘industry’ are large. BSc. to shed virus and to infect other birds by either vertical or horizontal transmission.Experience has shown that the productivity of rehabilitated birds is poor and an ever present risk of disease. considering the low level of productivity to high level of investment that has taken place. MSc.g. Health surveys show that wild houbara bustards are not naturally exposed to the viral diseases that are common in captivity. Bailey. central Asia) is both logistically complicated and can be controversial. the integration of any birds that have passed through the illegal trade into large breeding projects cannot be recommended. BVSc. avipox. because wild populations are declining and obtaining permission to conduct responsible egg collection in range states (e. Appropriate protocols will need to be developed to screen captive bred birds before they are released back into the wild. This may explain the apparently high susceptibility of smuggled birds to infectious diseases. the larger breeding centres will need to improve co-operation and develop common research goals. reovirus. Projects will also need to develop health screening standards for birds that are exchanged between projects. adenovirus. It is getting harder to obtain birds from source 3. The health issues surrounding the production. but latently infected birds. Viral diseases (e. because of inability to treat infections.ae 30 . The best sources of healthy breeding stock for captive projects are from sources 3 and 4. Ultimately. exists. AUTHORS ADRESS T. the lack of knowledge concerning recovery from infection and the potential for recovered. so that research funding is prioritised and existing and new problems are solved. Cert Zoo Med.g. in spite of expensive quarantine and health screening. avian leucosis) represent the clearest health threat from rehabilitated houbara incorporated into breeding programmes. Consequently. Projects therefore need to initiate genetic management to manage their founders and to investigate new ways of collecting new genetic material.bailey@dfh. MRCVS. utilisation and ultimately domestication of houbara bustards in the Middle East are complex and challenging. The founder breeding stock of the major breeding projects were all derived from eggs collected from the wild and were hatched in captivity and hand-reared. PhD Dubai Falcon Hospital PO Box 23919 Dubai / United Arab Emirates e-mail: tom. PMV-1. may include ataxia. R. and/or passage of undigested food in the feces indicating a malabsorptive or maldigestive disorder (GREGORY. probably as a result of the widespread importation and shipment of birds to satisfy the demands of the pet trade. especially the nerves that supply the muscles in the proventriculus and other digestive organs including crop. Athens. 1995). INTRODUCTION Proventricular dilatation disease (PDD) is used to describe an inflammatory response characterized by the accumulation of lymphocytes and plasma cells in the central and peripheral nervous systems. DVM. BS ABSTRACT Proventricular dilatation disease (PDD) continues to cause morbidity and mortality among companion and aviary birds. S. C. Subsequently. DVM. abnormal head movements. however. W. constant or intermittent regurgitation. which may occur in addition to or independent of gastrointestinal signs. 31 . Ritchie. K. To evaluate issues related to the natural transmission of the PDD agent. they should be considered psittacine or nonpsittacine birds susceptible. DVM. Field observations have been used to suggest that the etiologic agent of PDD can be spread through horizontally and vertically. breeding pairs of cockatiels in which at least one individual of each pair was histologically positive for PDD were placed in enclosures to facilitate breeding and both the adults and their offspring were followed to determine the rate of transmission in the population. MS. Pesti. USA EPIZOOTIOLOGY OF PROVENTRICULAR DILATATION DISEASE IN BREEDING COCKATIELS B. the importation of psittacine birds or their eggs into any region with indigenous Psittaciformes must be considered extremely risky. PhD. 1995). College of Veterinary Medicine. D. PhD. 1995. Central nervous system signs associated with PDD. This disease was first discussed in the late 1970‘s in birds imported into the United States and Germany (GREGORY. Latimer. Ard.Emerging Diseases Research Group. Gregory. Given the severe nature of PDD and its potential to affect a wide range of bird species. weight loss (with or without decreased appetite). 1991). M. University of Georgia. There is no reference to spontaneous disease in free-ranging psittacine birds. GERLACH. PhD. The most common clinical signs of PDD include depression. a PDD epornitic has been occurring in psittacine birds in North America and Europe. seizures and proprioceptive or motor deficits (GREGORY. ventriculus and small intestine. however. GREGORY et al. 1997). 6 and 7). 4. 1987. 1999. The absence of disease in contact controls suggests that the etiologic agent of PDD is not readily transmissible or requires a specific route of inoculation that was not favored by the experimental conditions. avian polyomavirus and avian encephalitis virus (GREGORY. Adenovirus-like particles were demonstrated within intranuclear inclusion bodies in one affected bird. and PDD has been shown to occur throughout the United States. coronavirus and reovirus have been detected in tissues. 32 . 2002). 2004). Clinical changes in experimentally infected birds vary from acute onset of a combination of central nervous system and gastrointestinal signs. secretions or excretions from birds that have been either histologically positive for PDD or with gross distention of the proventriculus. Antibodies to this virus could be detected using a virus specific ELISA. EXPERIMENTAL and NATURAL INDUCTION of PDD Exposing susceptible psittacine birds to a tissue homogenate derived from affected birds can experimentally induce the lymphoplasmacytic ganglioneuritis that characterizes PDD (Gregory.. Canada and Europe. 2. Similar inclusion bodies have been described in the nerves of pigeons with paramyxovirus infections (GERLACH. GRUND et al. enterovirus. none have been proven to be the etiologic agent (GREGORY. 1995). to induction of only gastrointestinal signs that were first noted 3 months after inoculation. Pacheco‘s disease virus (an avian herpesvirus). even though EEE virus occurs primarily in the eastern portion of the United States. 1991. None of these viruses have been consistently demonstrated in all birds with confirmed PDD. GERLACH. multiple etiologies have been proposed for PDD. 1999). followed by death within 11 days of inoculation. 1991). MANNL et al. however. Paramyxovirus-like viral particles were demonstrated within inclusion bodies located in the neural cells of the spinal cord and in visceral nerve ganglia of another bird. antibodies were not detected using standard hemagglutination-inhibition assays available for paramyxoviruses (GRUND et al. Experimentally infected African grey parrots either died soon after inoculation or seroconverted and shed virus with morphologic characteristics suggestive of paramyxovirus in their excrement (GRUND et al. Alternatively. Birds with PDD have been shown to lack detectable levels of antibodies to paramyxovirus (serotypes 1. Experimental and epizootiologic findings suggests that EEE virus is not the cause of PDD (GREGORY et al. 3. Using electron microscopy. GRUND et al. This finding was used to suggest that PDD may be caused by EEE virus (GASKIN et al. et al. The disease in these neonates was termed avian viral serositis. 1995. A paramyxovirus related to Hitchner B1 was recovered from birds with PDD. 1991). 1997. unpublished data. An eastern equine encephalomyelitis (EEE) virus was recovered from neonates with abdominal distention from an aviary with a history of PDD. 1991.ETIOLOGY of PDD Since its initial description in the late 1970‘s. it could be speculated that the unaffected contact controls were already immune to infection. GASKIN et al. viruses with morphologic characteristics suggestive of paramyxovirus. 1999). “Buddy” Steffens. any immunity passed from a hen to her chicks is only transient. both those that received the virus containing inoculum and the birds that received the control inoculum were derived from the same initial group of research birds. GREGORY CR (1995).However. 273 – 81. 33 . all birds. 31: 214 – 21. Neuropathic gastric dilatation in Psittaciformes. GREGORY CR.a 4 year study on clinical case history. NIAGRO FD. Adults and their offspring were monitored for clinical changes suggestive of PDD and birds that developed morbidity were euthanized and tissues were histologically evaluated. 2. the Stop PDD Challenge and the Kenosha Exotic Bird Club. FL. diseased adults can produce clinically normal offspring. HOMER B. Wingers Publishing. a gifted and cherished scientist. microbiological and virological results. chicks produced by positive parents are susceptible to disease. 3. REFERENCES 1. In: Ritchie BW (ed): Avian Viruses: Function and Control. 5: 27 – 34. These findings suggest that a lack of successful transfer of the virus from an infected to an uninfected bird was more likely than some specific immunity in all of the contact control birds. GASKIN JM. Veterinary Medical Experiment Station. MANNL A. A newly recognized syndrome of psittacine birds. Lake Worth. 4. To evaluate the epizootiologic characteristics associated with naturally and experimentally induced disease. ESKELUND K. J Avian Med Sur 1997. Riverbanks Zoological Park.L. bird clubs and veterinarians have also made important contributions. Hundreds of aviculturists. Preliminary findings in avian viral serositis. Investigations of eastern equine encephalomyelitis virus as the causative agent of psittacine proventricular dilatation syndrome. W. 1995:439-448. LEIPOLD R. J Assoc Avian Vet 1991. diagnosis. Zoo Atlanta. LATIMER KS. ACKNOWLEDGMENTS In memory of Dr. analysis of species. 11: 187 – 93. Most offspring were allowed to mature without interference while some were experimentally inoculated with tissue homogenates known to induce PDD. All of the experimentally infected birds were susceptible and none of the contact control birds developed disease. the severity of histologic lesions do not correlate with chronicity or death and the period from exposure to the suspect PDD agent to development of overt clinical signs can be more than a year. Proventricular dilatation disease. 5. Major sustained contributions that have made this work possible have been provided by the UGARF Animal Health Fund. breeding pairs of cockatiels in which at least one individual of each pair was histologically positive for PDD were placed in a closed indoor room and provided nesting boxes. Macaw wasting disease . GERLACH H. Proc Europ Chap Assoc Avian Vet 1991. epizootiology. colleague and friend. GERLACH S. Results of this study confirm that some birds can be in direct contact with PDD positive birds for prolonged periods without developing disease. et al. Avian Dis 1987. 49: 445 – 51. GELDERBLOM HR.6. GRIMM F. Avian paramyxovirus serotype 1 isolates from the spinal cord of parrots display a very low virulence. GRUND C.uga. 19 – 23. GA 30602 USA e-mail: britchie@vet. Proc Assoc Avian Vet 1999. Ritchie. WERNER O. Serological studies on persistent PMV-1 infection associated with PDD. 7. GRUND CH.edu 34 . et al. KOSTERS J. AUTHORS ADRESS B. et al. W. PhD Emerging Diseases Research Group College of Veterinary Medicine University of Georgia Athens. DVM. J Vet Med B Infect Dis Vet Public Health 2002. Main European Association of Avian Veterinarians Conference TEXTS . . They became extinct as a breeding species in the United Kingdom (UK) in 1832 probably as a result of hunting.R. Osborne BSc DPhil. Genetic studies indicate that Britain’s bustards probably belonged to the central European group and that restocking should not use birds from Iberia. D. Kapranova. artificially incubating them and transporting chicks to Britain. Pilot studies on arthropods in long grassland suggest that their density is sufficient for chick-rearing but the precautionary creation of additional food-rich areas among arable crops is recommended.Waters. A set of haematological normal values for captive juvenile great bustards will be proposed. 37 .Mortality ABSTRACT Great bustard eggs were taken from disturbed nests in Russia. It will also discuss the disease syndromes seen and health problems encountered from incubation to release. Khrustov KEYWORDS Great bustard . United Kingdom VETERINARY ASPECTS OF THE GREAT BUSTARD (Otis tarda) REINTRODUCTION PROJECT IN THE UK J. This paper will discuss the disease screening protocol devised for this project and the results gained. agricultural change and inclement weather. A. Only Russia has sufficient birds to supply a reintroduction project and losses there through nest devastation are high.Strathmore Veterinary Clinic. CertZooMed. the project should have zero detriment to the donor population (OSBORNE 2005). Andover. A. Fraser BSc. T. 1 INTRODUCTION The great bustard (Otis tarda) is a globally threatened species needing conservation action across Europe. MRCVS.Otis tarda . long grassland for feeding and adjacent arable land for nesting. Chitty BVetMed.Disease surveillance – Reintroduction . By rescuing eggs. The Plain combines short grass areas for lekking. Suitable habitat exists in pockets across England and especially on the Salisbury Plain where a large area is protected for military training and conservation purposes. These were hatched in captivity and chicks transported to the UK for reintroduction to the wild. The factors that caused the loss are no longer thought to operate. P. Hants. They were then moved to soft pens prior to release onto the Salisbury Plain. one dead bird did have flagellates and trematode eggs in the gut. Chick problems. Egg mortality. c.2 MATERIALS AND METHODS Eggs of the great bustard have been taken from disturbed nests in Russia and incubated since 1982 (PONOMAREVA 1983). Most common was failure to retract the yolk sac followed by infection. the death of one bird immediately post-transit and three others within three days of arrival. Of those that died in the UK. For the release site a large open pen was constructed. Naturally there were concerns over entry of disease into wild and domesticated birds in the UK. Thirty chicks of 33-50 days of age were shipped from Russia to the UK prior to spending 28 days in licensed quarantine. chicks have been allowed to imprint on the human rearers. For the purposes of the reintroduction project this process has been altered to prevent this imprinting. In addition to compulsory testing for influenza and paramyxoviruses during quarantine. This was planted with game cover crops to provide a basic plant diet and to attract invertebrates for the bustards. Delays in obtaining export visas in Russia meant that many of the birds were too old to travel. Similar crops were planted around the pen and at other sites near the release site. Parasitology. 38 . but success at reintroducing these chicks to the wild can be assumed to be poor (OSBORNE 2005). Faecal tests. 3 RESULTS 3. “Angel wing” was also seen frequently but was easily corrected using wing bandages.1 In Russia a. Methods used are similar to those used in the rearing of the whooping crane (Grus americana). probably coinciding with moving. Therefore the pool of birds from which the reintroduction birds could be chosen was much reduced. performed at 2 and 10 days post-arrival a. All birds were dewormed using fenbendazole and praziquantel. b. This was to allow certain guaranteed feeding sites and to “regulate” the dispersal of the birds. There were two deaths due to malposition. three had a bacterial hepatitis (one possibly linked to previous yolk sac infection) and one had a ventricular foreign body and rupture. In previous years. Post-mortem examination of those birds that died in Russia was not possible. 3. This did not reveal any parasites on either test. This may explain the deaths of two birds in transit to Moscow. Most showed early embryonic death.2 In UK: quarantine (1-28 days post-arrival) 1. However. testing for other pathogens was also carried out. No influenza or paramyxoviruses were isolated. For future virological investigation. A suggested normal reference range was obtained for birds aged 43-60 days. Blood Tests. 3. At the time of writing seven of the twenty-two released birds still survive in the wild. b. Probably bacterial and probably secondary to yolk sac infection. also. Hepatitis. Of the birds that died. c. 2. Hepatitis. Dislocated elbow.1 Soft pens (29-43 days post-arrival). Chlamydophila PCR. b. Swabs were taken from trachea and cloaca on arrival for storage and future testing.2 Post-release. None detected.b. d. c. 3. Three birds died the night after the first handling session 2 days postarrival. Serology. Therefore the period spent in soft pen was shortened so birds could be released earlier. Bacteriology. c. Wing amputated. performed at 2 and 10 days post-arrival a. Hepatitis. Blood Parasites. The risk of injury meant that predator awareness trials could not be performed. Haematology. Two injured birds (humeral fractures) have had wings amputated and are maintained in captivity. flagellates and trematodes. Deaths. Problems were associated with the age and size of birds following delays prior to export from Russia. Pre-existing gizzard foreign body. probably bacterial. Issues surrounding this include source of infection and zoonotic aspects. Male bird. d. 2. causes of death were attributed to trauma (10 birds) and predation (5 birds). Paramyxovirus. Virology. Probably bacterial. Ill on arrival at Heathrow airport and died that night. None detected. e. Four birds were positive for this organism. No pathogens were detected. 3. Biosecurity in the rearing unit will be improved. All birds that died 39 . e. Serum storage. a. Two birds were injured: Fractured humerus. No obvious problems detected and ranges similar to other birds. Four birds died in quarantine (see above) 1. Acid Fasts. Retained. Male bird. Cell storage for future DNA analysis and/or sexing. influenza and avipox virus serology was negative. Oryx 2005. Post-mortem examinations showed no sign of infectious disease. 6 CITATION INDEX 1. Andover. London Road. The disease screening programme will be continued for these with some modifications. SP10 2PH. Chitty BVetMed. OSBORNE PE. catching. Germany. Symposium uber die Grosstrappe (Otis tarda). Tanya Osborne and other members of the Great Bustard Group for the hard work in record keeping. 4. Over the next 5-10 years further groups of birds will be re-introduced from Russia.com 40 . Saul Cowen.had fed and were in good body condition at time of death. disease and mortality levels have been within acceptable levels for this type of project. The screening procedures appear to have indicated that birds being reintroduced are unlikely to constitute a serious health risk for indigenous UK birds. 4 DISCUSSION To date.pipex. handling and maintaining the birds in captivity. AUTHORS ADDRESS J. 2. Karen Waters. CertZooMed. 5 ACKNOWLEDGEMENTS The authors would like to thank Tom Bailey MRCVS. Dr D Welchman and colleagues at VLA Winchester for performing autopsies on dead birds in quarantine. 1983. MRCVS Strathmore Veterinary Clinic. Die Restitution naturlicher Populationen der Grosstrappe (Otis tarda) in der USSR. 39(1): 1 . Eberswalde. R. Dr Ruth Manvell and Chris Davis MRCVS for their help in designing the disease screening protocols. Hants. Key issues in assessing the feasibility of reintroducing the Great Bustard (Otis tarda) to Britain. UK Email: xmo32@dial. Removal of fences and marking of those that cannot be removed will now be carried out in the vicinity of the release pen. Medlab Laboratories for clinical pathology work. observing.8 PONOMAREVA TS. Chris Davis MRCVS and Kate Chitty MRCVS for assistance in treating and sampling birds. Botulism . marshes. seeds and the leaves.Crested screamer . They are primarily herbivorous. 1 INTRODUCTION Crested screamers are one of three species of screamer in the order Anseriformes and family Anhimidae. M. rivers and lagoons.Proventricular impaction ABSTRACT This paper discusses the management and medical conditions of crested screamers. They have no uncinate process. Barrows BSc BVMS CertZooMed MRCVS. Crested screamers are widespread in South America occurring from Bolivia. Hartley BVetMed MapplSc (Wildlife Health) MRCVS. Pittman CVT.Avian gastric yeast . Chauna torquata in captivity. M. The horned screamer Anhima cornuta is the sole member of the genus Anhima. Screamers are large birds weighing from 3 to 5 kg when adult and have several interesting anatomical and physiological characteristics.Chauna torquata . avian gastric yeast Macrorhabdus ornithogaster and proventricular impaction. although they will take some invertebrate prey.Diseases . ingesting grasses. KEYWORDS Birds . M. J. especially when chick rearing (AZA WATERFOWL TAG 2004). while the near threatened black-necked screamer Chauna chavaria and the relatively abundant crested screamer belong to the genus Chauna. flowers and roots of aquatic plants.Johannesburg Zoo South Africa DISEASES AND MANAGEMENT OF CAPTIVE CRESTED SCREAMERS (CHAUNA TORQUATA). Uruguay and Brazil south to Argentina and inhabit a variety of wetlands including swamps. Medical conditions discussed include botulism caused by Clostridium botulinum Type C toxin. These large South American birds are members of the family Anhimidae in the order Anseriformes and have several unusual anatomical and physiological characteristics. a lack of feather tracts or apterae and only a few developed lamellae on the 41 . used as a defence mechanism (PERRINS 1990). Feed dishes should be placed on solid mats so that when food spills over the side the chicks are not ingesting substrate. 2 DISEASES OF CAPTIVE SCREAMERS 2. Gastrointestinal impaction Gastrointestinal (proventricular) impaction is the most common and significant disease of captive screamers and is a major cause of chick mortality. weakness progressing to recumbency. Clutch size varies from 2-7 (DEL HOYO et al. 1992) and both parents incubate the eggs for 42 to 45 days.1. Preventative measures should include paying attention to the formulation of chick diets. Diet and husbandry. Norgine Ltd) and a maltodextrin and protein concentrate solution (Critical Care Formula. The chicks are precocial. Clinical signs include weight loss. The moult is gradual unlike most other anseriformes. Some collections regularly lose 50-75% of chicks to impaction and another author gives a general figure of 48% overall chick mortality within the first 30 days (AZA WATERFOWL TAG 2004). Impaction has been seen with inappropriate dietary sources of fibre such as alfalfa hay and with substrates such as sand and grit. However a combination of intravenous and oral fluids. however cases have been seen in juveniles almost a year old. They have large toes with vestigial webbing at the basal portion and both males and females have sharp carpal spurs. The intestines may also be impacted or may show gaseous distension. Diagnosis is straightforward from standard lateral and ventrodorsal radiographs which typically show a massively distended and impacted proventriculus. using plenty of water to hydrate pellets.inner surface of the bill. Chicks are generally affected within the first two months. dehydration and dyspnoea. The proventriculus is large and sacculiform and the liver is predominantly on the right hand side so that on radiographs a symmetrical liver shadow is not seen. Plenty of exercise is also important for chicks. and keeping screamers on a natural grass substrate rather than sand or loose soil. antibiosis. They are also said to have the most pneumatic bones of any bird species (AZA WATERFOWL TAG 2004) and can soar on thermals at great heights. They have a well developed system of air sacs beneath the skin which can feel on palpation like subcutaneous emphysema or oedema and can make jugular venepuncture difficult. in particular substrate type are important aetiological factors and the presence of avian gastric yeast Macrorhabdus ornithogaster in several of these cases may also be significant. Vetark UK) has proved successful. Screamers are named for their loud honking vocalisations and can live for up to 35 years in captivity. Treatment is often unrewarding especially in young chicks. cisapride and gavage with polyethylene glycol with electrolytes (Movicol. 42 .1 Non-infectious diseases 2. They can also swim but are mainly terrestrial.1. Treatment was mainly supportive and involved initial gavage with 20ml/Kg warm electrolyte solution (Darrows solution. suggestive of an acute shock reaction. including the caecum. Intestinal contents tested strongly positive with the mouse toxicity test for C. Botulism Like other waterfowl. On day two his oral discharge had decreased but he remained dull and inactive. neck and head. The male screamer was treated in situ in his aviary so that he could continue to feed the chicks that the pair was rearing. although interestingly in an outbreak at the authors’ zoo where the toxin was waterborne through a series of adjacent aviaries with an interconnected water source. She was much the same by day three.5ml C. wings.2. although she was drinking. able to walk. Histopathology revealed mild to severe congestion of several organs. lactated Ringers solution (20ml/kg SC to the male and 5ml/Kg IO per hour to the female). other than a haemorrhagic appearance to the caecum.3. however by day three he was much improved and the treatment was discontinued. Introduction of non-related adults should be done gradually in neutral territory 43 .1. They were gavaged with ground up waterfowl pellets three times daily. She was kept in a sling and physiotherapy carried out on her legs. SA) containing a probiotic (Avipro. screamers are very susceptible to botulism caused by Clostridium botulinum Type C toxin.2mg/ kg SC and enrofloxacin at 15mg/kg IM q12h. The adult male also displayed general paresis. On post mortem examination. a bilateral clear ocular discharge and a thick mucoid oral discharge. ivermectin at 0. They were also given a single dose of 0. Kyron Laboratories Pty Ltd. Since this outbreak our screamers have been vaccinated annually with a single dose of 1ml C. She could move her legs slightly but at times was unable to lift her head. Clinical signs included typical flaccid paralysis of the legs. Trauma Trauma has been reported as the most common problem of adult screamers (AZA WATERFOWL TAG 2004) and may result from intra or inter-specific aggression. He went on to successfully rear two of the four chicks.2. UK) and activated charcoal. botulinum antitoxin (Bio Onderstepoort) intravenously. unable to eat and still producing a thick oral discharge. On day seven she started regurgitating food after tube feeding and she died on day eight. She started eating a little on day four and was moving her legs more but did not show any further signs of recovery. botulinum type C. A clear ocular discharge and a thick mucoid oral discharge were also seen. there were no significant gross lesions visible. botulinum vaccine (Bio Onderstepoort) given subcutaneously.1. The six year old female presented in good body condition but was weak and unable to stand with a thick mucoid oral discharge. He was however. The female Screamer was holding her head up on day two but was still unable to stand. none of four one month old chicks were affected even though both parents succumbed to the disease. 2. Vetark. 2.4. Mixed bacterial infections are found. The plantar surfaces of the feet are relatively thin and are easily damaged or bruised.1.7. 2.2 Infectious diseases 2. Digit trauma Digit trauma is particularly common in screamers and is most likely related to inappropriate substrate. Other non-infectious diseases Other non-infectious diseases reported in screamers include renal gout and oxalate nephrosis. Capture myopathy Capture myopathy has been reported in screamers after prolonged manual restraint. dressings to take pressure off the affected area and surgery as in other species. 2. Crested screamers are monogamous and territorial during the breeding season (SILVEIRA and FOWLER 2001) but are fairly gregarious during the rest of the year and have been kept successfully in family groups for up to a year and in single sex groups for longer periods. One juvenile screamer at the author’s zoo required amputation of three out of four toes on each foot after damaging the digits at the level of P3/P4. It was thought that this occurred after she had difficulty getting out of the concrete pool in her enclosure.5.1.1. The mode of transmission is likely to be the faecal-oral route.1.2.utilising a divider fence. Bumblefoot Pododermatitis is common in screamers as in other waterfowl.6. Avian megabacteria gastric yeast (Macrorhabdus ornithogaster) or Avian gastric yeast is a relatively common post mortem finding in screamer chicks and may occur concurrently with proventricular impaction. Again this can be prevented by keeping screamers on a natural grass substrate and utilising natural surfaces rather than concrete or abrasive materials for ponds. Diagnosis may involve a Gram stain of faeces or of a proventricular wash. and treatment involves husbandry changes. 2. 2. Clinical signs include weight loss and loose unformed faeces or diarrhoea. antibiosis. although false negative results are possible if affected birds 44 . particularly when forced to stand on concrete or abrasive surfaces and can follow on from the plantar trauma and ulcers mentioned above. A young male screamer developed a 90 degree flexion of his digits at P3/P4 at around 7 months of age and consequently developed ulcers on the dorsal surfaces and a bird hospitalised on rubberised flooring for treatment of gastrointestinal impaction developed plantar ulcers and worn nails on all digits.1. Medpet Ltd. A course of oral amphotericin at 10mg/kg bid for 4 weeks. Histopathology typically reveals marked koilin dysgenesis in the ventriculus. sometimes with atrophy of the muscle layers and presence of the typical large rod-shaped yeasts in both the ventriculus and proventriculus. avian gastric yeast may not always be pathogenic. In addition.2.2. repeated biopsies from both birds have revealed only secondary bacterial (Staphylococcus aureus isolated) and food particle contamination.are not shedding the organism at the time of the test.2. nutritional status. The commisure lesions in the female failed to respond to a week long course of systemic and topical antibiotics and debridement. Collections which have had few problems with their screamers generally keep them on grass with access to a natural lake or pond where they can graze on aquatic vegetation. In spite of the similarity of most of these lesions to pox virus. gastrointestinal candidiasis and fungal rhinitis. These lesions were first apparent at 3 months and 7 months respectively and have not regressed over the subsequent months. Treatment is likely to be palliative and as in other species (PHALEN and TOMASZEWSKI 2003. Suspected pox virus Two siblings at the author’s zoo have developed pox-like proliferative lesions at the commisures of the beak. 2001). One of the birds has also developed raised crusty lesions on his feet and at one point had a focal 1mm whitish plaque adjacent to his glottis. factors such as age. along with an avian probiotic (Entero-Plus. However an attempt should be made to treat it in screamer chicks especially where weight loss is present and because koilin dysgenesis may affect ventricular function and be a predisposing factor for proventricular impaction. especially in adult screamers. cryptosporidiosis. He also had severely deformed feathers. RAVELHOFER-ROTHENEDER et al. Electron microscopy also failed to reveal a viral aetiology. 2. 2. however the proventriculus is large and may appear dilated even in healthy screamers. In other species. Other infectious diseases: Other infectious diseases reported in screamers include Capillaria sp. proventricular dilatation may be seen on radiography or at post mortem examination. with or without proventricular impaction. SA) resulted in resolution of diarrhoea and weight gain in a one year old juvenile screamer with a positive faecal Gram stain. Captive diets are based around commercial waterfowl or gamebird 45 . intestinal coccidiosis. 3 DISCUSSION Many of the diseases reported in screamers are husbandry related and can be prevented by good dietary management and by keeping them on a natural grass substrate.3. immunologic status and the presence of concurrent disease may affect pathogenicity (LANGLOIS 2003). The foot lesions consisted of severe acanthosis and orthokeratotic crusts with multifocal areas of moderate heterophil infiltration and degranulation in the dermis with associated overlying epidermal ulceration and serocrust formation. ENGELHARDT H. AZA WATERFOWL TAG Screamers In: Proc of Waterfowl Husbandry Workshop.111. 5 ACKNOWLEDGEMENTS The authors would like to thank the staff of Professional Zoolife Consultants and Johannesburg Zoo as well as Drs Thelma Meiring and Liza Du Plessis of Vetpath and Dr Emily Lane for carrying out histopathology on several of these cases. Investigation into the identification. 2. International Council for Bird Preservation: Headline Book Publishing PLC 1990. Spain: Lynx editions 1992. They have been kept successfully in mixed exhibits with a variety of species including Leopard tortoises. Ames: Iowa State University Press. especially outside of the breeding season. The definitive guide to birds of the world. 4 CITATION INDEX 1. 85 . 4. 2001. Swans). Screamers are usually kept as pairs but can be successfully kept in groups. In: FOWLER M and CUBAS ZS (eds): Biology. PERRINS C. Megabacteria – taxonomic classification and significance as pathogen in various bird species. Tenerife 2003: 79 – 83. Chicks are usually pinioned at around 3 days of age and a three foot barrier is suitable for containing pinioned birds.za 46 . ELLIOT A and SARGATAL J (eds): Handbook of the Birds of the World. 103 . flamingos and psittacines. 7. RAVELHOFER-ROTHENEDER K. Vol 1 Ostrich to Ducks. SILVEIRA L and FOWLER M. Chicks may require an increased percentage of protein (AZA WATERFOWL TAG 2004) and one collection routinely adds probiotics to the chick diet. 6. Proc Euro Assoc Avian Vet.114.pellets with a variety of chopped greens. camelids. 3. tapirs. Santa Barbara 2004. Proc European Assoc Avian Vet. WOLF O et al. PHALEN D and TOMASZEWSKI E. Order Anseriformes (Ducks. Medicine and Surgery of South American Wild Animals. AUTHORS ADDRESS M. LANGLOIS I. Munich 2001: 189 . Geese. 696.org. DEL HOYO J. physiology and diseases of the avian proventriculus and ventriculus.Barrows. The anatomy. detection and treatment of the organism formally known as megabacterium. In: HERNANDEZ-DIVERS and HERNANDEZ-DIVERS (eds): Veterinary Clinics of North America Exotic Animal Practice Internal Medicine 6 2003. The Illustrated Encyclopaedia of Birds. 76. 5.190. BSc BVMS Cert ZooMed MRCVS Professional Zoolife Consultants PO Box 956 Houghton 2041 Johannesburg South Africa Email: Vet@jhbzoo. capybara and several bird species including other waterfowl. Barcelona. except for vagrants. two are natives of North America (the trumpeter swan and the whistling swan) and three are natives of Eurasia (the mute swan. KEYWORDS Swans – Grounded – Unable to stand – Examination – Diagnostics – Treatment. Free-living populations of non-native species do exist. clinical examination and assessment of such swans. The 47 . Further diagnostic tests will include blood sampling for haematology and biochemistry. having arisen as the result of introduction programmes for sport. weather conditions. crash landing. A healthy swan is unlikely to allow itself to be approached and will usually stand and make some sort of response. weighed and condition scored. 1972). the coscoroba swan and the Australasian black swan). however. The term “downer swan” has been borrowed from cattle medicine to denote the swan that does not stand or respond when approached. Bewick’s swan and whooper swan). being indigenous to every continent except Africa and Antarctica (SCOTT et al. lead poisoning. there is no movement between continents and none between the two hemispheres (SCOTT et al. 1972). The bird should be identified. A detailed examination will help establish a list of differential diagnoses. Cousquer BSc(Hons). The contact details of the finder should always be recorded. three are natives of the southern hemisphere (the South American black-necked swan. 1 INTRODUCTION Swans have an almost worldwide distribution. power line injuries. These will include fishing tackle problems. egg peritonitis and migration exhaustion. time of day and breeding history. Of the eight species and subspecies. radiography and endoscopy. This paper will review the history taking. United Kingdom CLINICAL APPROACH TO THE DOWNER SWAN G. Such swans should be investigated.Private Practitioner. Clinical examination will look for signs of acute versus chronic disease. Most of the populations in the northern hemisphere are migratory but. chronic disease (tuberculosis and aspergillosis) and more unusual presentations such as parasitism. MRCVS. ABSTRACT It is not uncommon for swans to be found on the ground. CertZooMed. flight paths. BVM&S. History taking will include the recording of key information such as the location. ornamental and other purposes. The domestication and farming of the mute swan in Britain dates back to medieval times as does the bird’s royal status. all unmarked swans on open or common land remain the property of the Crown to this day (SCOTT et al. 3 ASSESSMENT AND CLINICAL APPROACH The assessment and clinical approach to the downer swan will. attention should also be paid to the following: . together with a wide range of underlying aetiological problems can make the investigation of such cases diagnostically challenging.mute swan (Cygnus olor) is common in many parts of Europe and Asia. The keeping of detailed and accurate records is not only good clinical practice but can greatly assist future retrospective studies.Breeding history. Admission details should include the time and date of admission. particularly in the British Isles.Recent observations 48 . 2 DIFFERENTIAL DIAGNOSES The range of possible differential diagnoses is expounded in Table 1. where and when found and the “reason for admission”. In northern Europe. indeed. clinical examination and subsequent diagnostic tests. Austria and Turkey.Habitat. a standardised admission sheet will ensure that nothing is omitted. Holland. This paper describes the clinical approach to these patients. .Local flight paths.Type of waterway . In addition. They fall broadly into traumatic and non-traumatic aetiologies. 1972). identifying rings or markings. It occurs less abundantly in Belgium. . . Switzerland.Response when approached . The latter include a range of conditions that have a predominantly neurological presentation but also include chronic disease and miscellaneous conditions such as egg peritonitis. the mute swan’s present status and distribution has been greatly influenced by introductions and a long history of domestication. Many of these birds will present as weak or unable to stand. age. it is essential that the finder’s contact details be recorded to allow them to be contacted at a later date. Poland and southern Sweden. northern France. the mute swan is a common bird on British waterways and frequently presents to wildlife hospitals and rehabilitation centres (COOKE 2003).Prevailing weather conditions. should the need arise. species. drawing on the author’s extensive experience to provide an illustrated review of this subject and a framework for the clinical approach to the “downer swan”. Use of an OS (Ordnance Survey) map (1:25000 or 1:50000) can prove helpful when identifying and recording the exact location at which the casualty was found. for the purposes of this article. Germany. Limited funds and facilities.Proximity to power lines.1 Anamnesis On admission it is essential that the opportunity be taken to record all key information. . and particularly in the British Isles. grieving and migration exhaustion. be broken down into history taking (anamnesis). Today. 3. In the case of a swan. . Normal weights will vary between 7. adopt an imposing threatening posture with its wings outstretched. Failure to stand is likely to be significant and should be noted and investigated further 3. convexity is scored a 2. This is particularly important where swans have crashed into power lines as electricity companies have a responsibility to respond to any such problems. alert and responsive. Concavity of the pectoral muscles is scored 0. Failure to lift up the head with the head remaining kinked over the back is suggestive of lead poisoning. This should be noted if one of the pair has recently been killed. The nature of the habitat and the type of waterway can be relevant: certain waterways. In cases of head trauma. A condition score of 0 to 3 should be ascertained by palpating the pectoral musculature and assessing the prominence of the keel. The body weight and condition score should be used together to assess the condition of the bird. As the bird is approached and stressed intention tremors of the head may become more obvious suggesting cerebellar damage or other CNS pathology. When approached a healthy swan will usually stand and.2 Clinical examination Attitude and appearance: The swan should be observed from a distance before a systematic head to toe examination is performed. Body weight and condition score: The swan should be weighed and condition scored. thus disappearing from view (CRACKNELL 2004). a swan flying in fog or into the setting sun can fail to see an obstacle and crash into it. flattening of the muscles is scored 1. It is normal for a clump of weed to accumulate at the base of the 49 . Condition scoring can be difficult in deep barrel-chested birds such as swans and is best done adjacent to the keel. The bird’s ability and willingness to stand when approached should be noted. Knowledge of local flight paths and an accurate record of all such incidents will help analyse trends in such incidents and allow preventive measures to be taken. these areas. and rounding that makes palpation of the keel difficult is scored a 3. particularly inner city canals and lakes. This can make fishing tackle related problems particularly common. for “chinstrap injuries” as fishing line will often be found to have cut into the tissues. Head and neck: A detailed examination of the head can be very informative: Examination of the oral cavity should be thorough to ensure that the presence of translucent fishing line under the tongue is not missed. The commissure of the beak should be examined. or that have moved through.A detailed history can shed light on the possible reason(s) for presentation. especially if male. are heavily and often irresponsibly fished. If a swan has been on the ground for a period of time the surrounding vegetation may be flattened. bruising may be evident within the mouth and there may be bleeding from the choana. A bird in poor condition is likely to have been ill for some time or failed to thrive through other circumstances such as bullying or limited food availability.3kg and 10. The healthy swan will be bright. Breeding pairs can be badly affected by the loss of a mate. Certain waterways are also heavily contaminated with lead shot and clinical as well as sub-clinical lead poisoning will be common in swans from.6kg (SNOW and PERRINS 1998). or if the capture and removal of one of the pair for treatment is being contemplated. Thus. A necrotic smell may be present where fishing tackle injuries have resulted in tissue necrosis or food impaction. keel wounds are a common finding in grounded. Female swans will remain on the nest despite receiving injuries to the neck and legs. This should be differentiated from any inter-mandibular swelling arising as the result of fishing tackle injuries to the base of the tongue and inter-mandibular space. Body and wings: As with most waterfowl. arising as the result of bites. This involves the loss of all the wing quills. It is not unusual for dogs to attack waterfowl who are less mobile than passerines and therefore easier to chase. Soft tissue wounds are not uncommon. The ears should be examined for bleeding following head trauma. Parents normally moult at different times in order to better protect their brood. while male swans will continue to defend a nest despite incurring bite wounds to the leading edge of the wing each time they strike out (ROUTH 2000). oesophageal tears and other traumas. Traumatic head injuries may involve the eyes and many swans will present with long-standing unilateral. Deformities and injuries of the cervical spine are not uncommon as a result of in-flight collisions. The neck should be carefully palpated. Sadly. Cataracts are not uncommon in swans and such swans will need this taken into account if they are to be released. Such birds spend long periods sitting down and will fall heavily onto their keels through weakness. There may also be an accumulation of faecal material around the vent. The back of the head is often parasitised with lice (Mallophaga spp. The nares should be examined for bleeding. Whilst this condition renders the individual swan flightless and vulnerable to attack. Birds that have been grounded for a period of time are likely to have superficial soiling of the ventral abdomen. Evidence of diarrhoea should be noted as this may reflect lead poisoning. Food impactions of the oesophagus may reflect an obstruction or impaired crop emptying as the result of lead toxicosis. The keel should be carefully palpated and visually inspected for keel wounds. and even bilateral. botulism. it does not of itself render them unable to stand. Debilitated waterbirds tend to allow their centre of gravity to move forward so that their keel is scraped along the ground (COOKE. Sustained contact with the ground will result in localised ischaemia and ulceration. The development of a pressure ulcer is a progressive process that will eventually result in bone necrosis (COUSQUER 2003a. The breath should be smelt. either as the result of trauma or due to parasitism with leeches (Theromyzon spp. A swan poses a formidable adversary and will rear up and defend itself with its wings.). parasitism or bacterial enteritis. resulting in a visible and palpable distension of the inter-mandibular space. rendering the swan flightless and relatively helpless until the new ones have grown (SCOTT et al. 1972).) and the resulting feather damage should be differentiated from feather plucking that has arisen as the result of bullying. which can invade the sinuses in large numbers. power-line injuries. Leeches may feed from the palpebral conjunctiva. Consequently dogs need 50 . A full ophthalmological examination is indicated to establish the extent of any visual deficits. COUSQUER 2003b).tongue. ocular pathology. Ketone bodies can also be smelt on the breath of swans that have been unable to feed and are therefore in negative energy balance. Fishing line and hooks may be palpated within the cervical oesophagus. swans pass through an eclipse moult during the breeding season. 2003). Rupture of the cervical air sac can result in accumulation of air within the tissues of the lower neck that will feel crepitant on palpation. weak and emaciated birds. Electrical burns will occur where contact has been made between two wires. total protein. such as the VLA. 1972) and are likely to result in injuries to the neck and to other parts of the body. have to be sent away to an approved specialised commercial laboratory. Injuries will not only arise as a result of the initial impact on the line. The considerable associated tissue damage accompanying such electrical burns will dictate euthanasia in the majority of cases (COUSQUER 2003b). total white blood cell count and blood lead. For this reason it is essential that all bite wounds are thoroughly investigated and assessed. causing considerable damage. Fishing tackle will often cut into the lower leg. Blood for lead testing will. in the author’s opinion be blood sampled. Pododermatitis is common but is rarely likely to render a swan unable to stand and walk unless associated with a septic arthritis or tenosynovitis. Devitalised tissue will become cold and oedematous to the touch. These can be very debilitating and render a swan extremely lame. lead poisoning should always be suspected even if it be only at a subclinical level. Traumatic luxations of the femorotibial joint are probably sustained when the tibia is loaded and pushed proximal to the femoral condyles. Trauma to the foot when hitting a wire. PERRINS et al. The minimum data-base should include measurement of the bird’s PCV. before sloughing in a compartmentalised fashion. Legs and feet: Careful palpation of the legs is indicated to detect both soft tissue and orthopaedic injuries. Infection tracks readily between the fascial layers of the tail muscles and euthanasia may be the only option in many cases (COUSQUER 2003b). vehicle or other object in flight can produce injuries to the dorsal surface of the foot and lower leg (COUSQUER 2003b. Some of these injuries can be very severe and vulnerable to fly strike. total protein and total white blood cell counts in house. (2003) found that slightly over 60% of all casualty swans sampled had blood lead levels in excess of 1. Other common findings include septic arthritis of the inter-tarsal joint and especially of the metatarsophalangeal joint. delineating the tract along which the electricity travelled (ROUTH 2000). skin or musculature over a period of up to five days.21µmol/l. It is not uncommon for swans to present with a range of lower limb injuries following a crash landing.to attack from behind and it is perhaps for this reason that the tail and rump area is most often targeted (COUSQUER 2003b).3 Further diagnostic tests Blood sampling: All casualty swans should. however. While these results were necessarily biased towards sick and injured birds. Finances will often limit the scope of any investigation but it should be possible to obtain reasonably accurate PCV. In the “downer swan”. Some of this lead will be removed from the blood and stored in tissue such as bone and liver from where it can be released back 51 . 3. but also as a result of the subsequent crash-landing (ROUTH 2000). they reflect the fact that a significant proportion of these birds have been exposed to lead and may be suffering from the effects. either at a clinical or subclinical level. Collisions with power lines are common (SCOTT et al. The manifestation of electrical injuries can be insidious (ROUTH 2000) and all birds found adjacent to power lines should be thoroughly and repeatedly examined for signs of burns to the feathers. Chronic lead poisoning is likely to arise as the result of the absorption or the release of lead into the blood stream over a relatively long period. ROUTH 2000). egg peritonitis and chlamydiosis. pectoral girdle and cranial airsacs (cranial thoracic and clavicular). The radiographs should be assessed carefully to glean the maximum amount of information: Are they symmetrical and of good diagnostic quality? A symmetrical radiograph will allow comparison of structures on the left and right sides of the bird. Further investigative blood work is indicated where abnormalities are found or the clinical picture demands further investigation. Lateral radiographs can be taken in the conscious swan but require the swan to adopt an unnatural position. in - 52 . A differential white blood cell count will prove very useful in such cases and will allow an assessment to be made of any response to therapy. difficult as detail of the left hip is usually obscured by the underlying ventriculus. the lower legs should be extended and tied down to prevent the swan from standing. caudal airsacs (caudal thoracic and abdominal). Finally. true of old weights that may have been weathered on the riverbed. The wings can then be extended away from the body and immobilised with sand bags. Is the picture an inspiratory or expiratory view? A max-inspiratory view will provide increased contrast and improved detail and is the gold standard. New lead weights are recognisable by their shape (split shot). are not fully understood. however. Is there a general loss of definition? This may reflect bleeding into or inflammation within the coelomic cavity. Is there any evidence of fishing tackle in the GI tract? The femoral cortices can be used to compare density of any material in the gizzard. Is there any evidence of air-sac disease? Sacco-sacco membranes should not be identifiable and there should be no filling of the air-sacs. The possibility that subclinical lead poisoning may underlie the downer swan’s condition should not be overlooked. “thoracic” oesophagus. thus calming the bird. Comparison of the hips is. Creatine kinase and aspartate aminotransferase levels will reflect the extent of myonecrosis and can be monitored to provide some indication of the progression of disease and rate of recovery. which is far more stressful for the bird. A view centred on the mid-scapular area and dorsal midline will provide information on the heart. Biochemical analysis can also be useful. Uric acid levels will be significantly elevated in cases of renal failure. It is clear that the metabolism of lead in swans and the means by which it causes the observed clinical signs. An elevated total white blood cell count should be investigated further to explore the possibility of chronic diseases such as avian tuberculosis and aspergillosis. The neck should be weighted down and the head covered. however. An elevated white blood cell count will also be found in relatively uncommon conditions such as cholangiohepatitis (PATTERSON-KANE and COUSQUER 2005). Is the proventriculus distended? This may suggest loss of GI motility. Lead weights are considerably more radio-dense than bone and grit. Radiography: Dorso-ventral radiographs are easily taken without the need for sedation or general anaesthesia. This is not. The action of the gizzard can. In this way it is possible to obtain a good quality symmetrical dorso-ventral x-ray: A view centred on the hips and dorsal midline will provide information on the digestive tract distal to the proventriculus.into the circulation following hepatocyte atrophy for example (WOBESER 1997). liver. hips and knees. they may assist in the removal of fishing tackle and are particularly useful in establishing the presence and extent of a tear in the wall of the oesophagus or proventriculus following removal of a fishing hook. or where endoscopy is not available. A fiveday course of treatment is advocated with blood lead levels retested on day eight after a two-day rest from therapy (ROUTH 2000). The liver margins should be distinct. Is there any evidence of kidney disease? An increase in kidney density can be seen when urates build up in the kidneys as the result of kidney disease or in the dehydrated patient. Consequently. Is there any evidence of liver pathology? An emaciated bird is likely to have a small liver whilst loss of the hourglass shape of the heart and liver suggests hepatic enlargement. Flexible endoscopes can be used to examine the digestive tract. Is there any evidence of joint disease? Stifle injuries are particularly common. This is performed under general anaesthesia. therapy may be initiated before a diagnosis is established. loss of definition may reflect tears and bleeding into the coelomic cavity. This should be born in mind as smaller lead particles have a greater surface area to volume ratio. however.- - time. treatment with sodium calcium edetate (NaCaEDTA) may be initiated. liver and kidney disorders. The patient will benefit from intensive intravenous fluid therapy and this will allow intravenous NaCaEDTA injections to be easily and aseptically administered. Additionally hospital staff will find it difficult to administer twice daily i/m injections aseptically in such a big bird. Is there any evidence of spinal injuries? Endoscopy: In many cases it will only be possible to confirm a diagnosis by visualising and biopsying lesions endoscopically. it may be necessary to recourse to an exploratory procedure. A blood sample should always be collected and submitted for lead testing prior to starting treatment. and the investigation of suspected avian TB. are more easily absorbed and therefore potentially more toxic. Intramuscular injections of NaCaEDTA should not be administered as they have the potential for causing considerable muscle necrosis (COOKE 2003) and may affect a bird’s flying ability and release prospects. Where intravenous access is not possible. Exploratory coeliotomy: In certain cases. reproductive disorders (e. 53 . based on clinical signs and/or the presence of lead shot in the gizzard. no substitute for an accurate diagnosis. Rigid endoscopy is ideal for the investigation of air-sac disease. result in new lead weights becoming polished and even breaking up.g. Where lead poisoning is suspected. egg peritonitis). Standard techniques are indicated and will allow a suspected diagnosis to be confirmed. the next best alternative is an aseptic subcutaneous injection over the calf. NaCaEDTA can be dosed at 35-50mg/kg i/v or s/c twice daily. Trial therapy: In certain situations. The drug is only licensed for intravenous administration. a “spike” should be placed in the bottle’s septum to allow the drug to be removed aseptically without introducing any needles into the bottle. Trial therapy is. Anaesthesia is induced using a xylazine and ketamine combination (ROUTH and PAINTER 1999). This will be the case when blood lead results are pending for example and a swan has presented with signs of acute lead poisoning that require urgent treatment. NaCaEDTA does not contain any antimicrobial preservative and is prone to contamination. Lead and other metals. PATTERSON-KANE JC. 32(2): 205 .231. World Wide Wounds. In: Proc Autumn meeting of the British Zoological Society 1999: 17 . com/2003/november/COUSQUER/avian-wound-management. 151 .html 4. PERRINS CM. November 2003b. Cholangiohepatitis and cholelithiasis in a mute swan (Cygnus olor). In Practice 2004. London: Michael Joseph Ltd. COUSQUER GO. WOBESER GA. 7. J Avian Pathol 2003. COUSQUER (2003b) and ROUTH (2000).worldwidewounds. 10. 9.html 3. Available from: www. Accepted for publication in the Veterinary Record.worldwidewounds. In: BSAVA Manual of Wildlife Casualties. August 2003a. SCOTT P. COOKE (2003). com/2003/august/COUSQUER/avian-wound-assessment. grebes and divers.245. 2. SCOTT and the Wildfowl Trust. and COUSQUER GO. CRACKNELL J.20. Private Practitioner 54 . World Wide Wounds. New York: Plenum Press 1997.4 TREATMENT The clinical approach outlined above should allow a diagnosis to be made and will facilitate the formulation of a therapeutic plan. P. 1972. Dealing with line and hook injuries in swans. 5. Available from: www. The use of intravenous ketamine and xylazine for induction of general anaesthesia in the mute swan (Cygnus olor): A review of 130 cases. ROUTH A. Waterfowl: swans. In Practice 2000. 1. Wound assessment in the avian wildlife casualty. 219 . 5 CITATION INDEX COOKE SW. Veterinary care of the mute swan. A Survey of lead levels in mute swans (Cygnus olor). 6. COUSQUER GO. In: WOBESER GA. Wound management in the avian wildlife casualty. 22: 426 443.212. COUSQUER GO and WAINE J. AUTHORS ADDRESS Glen COUSQUER BSc(Hons) BVM&S CertZooMed MRCVS. ducks. The Swans. Details of specific treatment protocols are available from CRACKNELL (2004).159. 26: 238 . 8. (ed): Diseases of Wild Waterfowl 2nd ed. Gloucester: BSAVA 2003. ROUTH A. geese. and PAINTER KS. Soft tissue trauma. Impactions. Classification Trauma Sub Classification Acute Aetiology Crash landing Key Findings Orthopaedic injuries. Hook damage to the beak or GI tract.Table 1: Differential diagnoses for the “Downer Swan”. Shotgun injuries. Septic arthritis especially of MTP joint. Degenerative joint disease Bumblefoot Neurological Intoxication Lead Poisoning Botulism Traumatic Blue green algae Spinal Injuries Head Trauma Infectious Other Chronic disease Miscellaneous conditions Bacterial Fungal Encephalitis Kidney Disease Avian Tuberculosis Aspergillosis Egg peritonitis Hepatitis / Cholangiohepatitis Bullying Broken heart (grieving) Migration exhaustion Parasitism Female birds. Crossbow injuries. Burns. Following loss of mate. Usually fatal. Hip luxations and arthritis. Flaccid paresis. UMN or LMN. Flown into power lines Dog bites Shooting injuries Fishing hook injuries Fishing line injuries Chronic All of the above if not identified early. Bite wounds to wing. Pododermatitis. Vestibular symptoms. “Cheese wire” injuries. Visual deficits. LMN signs. Airgun injuries. neck and tail. Bright green diarrhoea. Internal injuries. Whooper and Bewicks. Concussion. GI stasis. Ataxia. Often accompanied by tissue necrosis. Ataxia. Head and neck injuries. Gizzard worm (Amidostomum) 55 . Internal injuries and haemorrhaging. Head tremor. Weakness and anorexia. Stifle luxations and arthritis. Orthopaedic injuries. Respiratory distress. “Limber” (kinked) neck.Soft tissue trauma. Soft tissue trauma. Weakness. while overall prevalence in birds of prey was 10. Höfle DVM. JCCM. Prevalence of T. R. Asistencia Veterinaria sl.trichomoniasis ABSTRACT Avian trichomoniasis is a well known disease of the upper digestive tract of pigeons and other avian species caused by the protozoan parasite Trichomonas gallinae.Aquila Foundation. Gortazar DVM KEYWORDS Trichomonas gallinae . but among the woodpigeons sub-clinical effects on fat deposition and bursal size were observed. psittacine and 56 . gallinae and associated lesions. BOAL et al. from Bonelli’s and Imperial eagle nestlings and adults in the field and from 14 other species upon admission to the CERI (Centro de Estudios de Rapaces Ibéricas) were analysed for the presence of T. Villanúa. Blanco DVM. Prevalence of lesions was low. 1992). a disease affecting the upper digestive tract primarily in columbiforms (MEHLHORN et al.Passerine birds . Centro de Estudios de Rapaces Ibéricas CERI. gallinae results in sub-clinical effects due possibly related to a physiological expense for the host. ESVA. 1 INTRODUCTION The flagellated protozoan Trichomonas gallinae is the causative agent of avian trichomoniasis. M. Birds of prey. J. from mistnet captured passerines. 1998) and trichomoniasis has also been described in passerine. D. especially species that catch avian prey are also known to be susceptible to the disease (COOPER and PETTY 1988. Instituto de Investigación en Recursos Cinegéticos IREC.28%. C.Birds of prey . gallinae in woodpigeons was 43%. The preliminary results suggest that parasitation by T. Valboa. Spain TRICHOMONAS GALLINAE IN FREE-LIVING BIRDS OF PREY. samples from hunter-harvested healthy woodpigeons. T.Wood pigeons – Sub-clinical infection . The disease has recently emerged in some accipiter species as a factor in nestling mortality due to changes in the diet of these species. Host immune status appears to be an important factor in the development of clinical disease. To obtain further information on the epidemiology of the disease. gallinae could not be cultured from any of the passerines examined. PASSERINE BIRDS AND WOODPIGEONS (COLUMBA PALUMBUS) U. 2000. In an ongoing study we evaluate the prevalence and effects of T. Identification as T. In addition samples were obtained from five nestling and five adult Bonelli’s eagles in the field as well as a total of 20 Imperial eagle (Aquila adalberti) nestlings. incubated at 36. Overall prevalence of T. for culture samples were taken from the oropharynx.gallinaceous birds (COLE 1999).5º C and inspected daily for seven days for the presence of flagellated protozoa. Several authors have been concerned about the effect of the transmission of the parasite from reservoir to naïve hosts. In the rock pigeon (Columba livia) parasitism is generally sub-clinical and development of lesions is supposed to depend on immune status of the host and pathogenicity of the infecting strain (STABLER 1954.28% with prevalence ranging from 100 to 0% (table 1). while in the woodpigeons prevalence was higher among adults than among 57 . MATERIAL AND METHODS Samples were obtained from birds of prey. 3 RESULTS Prevalence of T. gallinae was analysed by culture. gallinae in the examined woodpigeons was 43% while the parasite was not cultured from any of the passerine birds examined. We sampled 199 passerine birds of 16 different species at two sites in central Spain in both winter and summer. HÖFLE et al. In clinical disease fibronecrotic lesions typically develop in the upper digestive tract although occasionally involving the respiratory tract. gallinae was achieved using morphology and PCR (HÖFLE et al. oesophagus and crop using sterile swabs and placed in sterile tubes containing 5 ml of CPLM medium supplemented with 10% foetal bovine serum and 10% antibiotic solution. as well as burdens of intestinal and external parasites were collected. passerines and woodpigeons. 2000). SAMOUR 2000). To date 107 birds of prey of 16 different species have been sampled upon admission to the CERI in central Spain. Each of the sampled birds was examined carefully for the presence of oropharyngeal or crop lesions and presence of T. In addition. REAL et al. 1998). gallinae among free-living birds of prey. gallinae among birds of prey sampled was 10. 2. COOPER and PETTY 1988). such as other endangered columbiforms after introduction of domestic pigeons from Europe into America (see STABLER 1954) or birds of prey (BOAL et al. In birds of prey a direct relation was established between the consumption of urban pigeons and the appearance of trichomoniasis in the nestlings of Cooper’s hawks (BOAL et al. Shortly. sinuses or ear (STABLER 1954. Most of the birds of prey in which the parasite was found were juveniles. and woodpigeons from central and southern Spain. 2004). but relatively little is known about the epidemiology of trichomoniasis in susceptible avian species. in the woodpigeons complete biometry and organ weights. passerine birds. 1998. Samples were obtained from 61 hunter harvested woodpigeons from two different locations in their wintering grounds in central and southern Spain. National Geological Survey 1999. The findings in the woodpigeons may provide evidence for the cost every parasite imposes on its host as defined for other host parasite systems (HUDSON and GRENMAN 1988). PALMA L and MELO P. GORTÁZAR C. 58 . J Wild Dis 1998. as in rock pigeons. Lake Worth: Zoological Education Network Inc 2000. REMPLE JD. PETTY SJ. p<0. It could thus explain that T. (eds): Field Manual of Wildlife Diseases. et al. p <0.87. (eds): Raptor Biomedicine III.77.1. and the parasite was cultured from half of the imperial eagle nestlings examined. Trichomoniasis in Cooper’s hawks from Arizona. 4 DISCUSSION Our results suggest that. COOPER JE.6. 3. MANNAN RW and HUDELSON. 5 CITATION INDEX 1. J. COLE RA.juveniles. REDIG PT. 2. sub-clinical infestations with T. ORTÍZ JA and KNISPEL B. C. In: FRIEND. 5. gallinae positive pigeons aggregate more at easy food sources and are more easily predated and thus propagating the parasite. COLE 1999). 50: 73 .19%) was significantly lower than prevalence of the parasite from culture. Similarly in the woodpigeons prevalence of lesions (2. BLANCO JM.05) and a significantly larger cloacal bursa (t-value =2. Department of Interior. European J Wild Dis 2004.02). gallinae do exist in birds of prey and woodpigeons and that development of disease depends on interaction between host immunity and strain pathogenicity as stated by other authors (STABLER 1954.52.206. as diagnosed from macroscopic lesions or typical clinical signs (table 1). U. HÖFLE U. HÖFLE U. In: LUMEIJ JT. Three of the culture positive Imperial eagles and one of the positive Bonelli’s eagle nestlings had clinical disease. T. Trichomonosis. M. No difference appeared to exist in prevalence between sexes. 34: 590 . Trichomoniasis in Bonelli’s eagle (Hieraaetus fasciatus) nestlings in South-west Portugal. However when comparing body condition and organ weights of parasitized and non-affected woodpigeons. 3. J Wild Dis 1988. gallinae.13. Trichomoniasis in free-living goshawks (Accipiter gentilis gentilis) from Great Britain. 4. However the observations could also be related to the effect of a subjacent pathogen or factor that predisposes woodpigeons to infection by the parasite. 45 . 201 . woodpigeons positive for T. Outbreak of trichomoniasis in a woodpigeon wintering roost. BOAL CW. KS.593. All of the Bonelli’s eagle nestlings and none of the adults were positive for T. and FRANSON. gallinae had significantly less body fat deposits (t-value= 2. gallinae and disease In the sampled birds of prey only very few of the culture positive animals had clinical disease.S. 24: 80 . 36: 64 .3 0 0 100 0 10.70. Supraorbital trichomoniasis infection in two saker falcons (Falco cherrug) Vet Rec 2000. MEHLHORN H.13005 Ciudad Real. Competition mediated by parasites: biological and theoretical progress. 8. 13: 387 . Culture results for T. gallinae and prevalence of clinical disease in birds of prey admitted to the CERI Species Scops owl Tawny owl Long-eared owl European Eagle owl Little owl Barn owl Marsh harrier Montagu’s harrier Peregrine falcon Common kestrel Lesser kestrel Goshawk Booted eagle Total n 5 10 8 13 12 10 7 9 3 18 10 1 1 107 Culture positive 0 0 0 0 2 1 3 2 2 0 0 1 0 11 Prevalence (%) 0 0 0 0 16.390. AUTHORS ADDRESS Ursula Höfle Instituto de Investigación en Recursos Cinegéticos IREC Ronda de Toledo s/n.28 Clinical disease 0 0 0 0 0 1 0 0 2 0 0 1 0 4 59 . Exp Par 1954. Trichomoniasis in a Bonelli’s eagle population in Spain. J Wild Dis 2000. REAL J. Atlas de Parasitología Veterinaria.uclm. 7.402. 3: 368 . SAMOUR JH. 146: 139 . 6.22 33.HUDSON PJ and GRENMAN J. 9. DÜWELL D and RAETHER W.140. Barcelona: Ediciones Grass. Spain Email: uhofle@irec. Trichomonas gallinae: a review. Trends Ecol Evol 1988. 10.67 10 42. 1992. STABLER RM. MAÑOSA S and MUÑOZ E.86 22.es Table 1. Veterinary University of Lyon. D. H. we set up a research program combining ornithological and serological approaches. birds are considered as the principal hosts of the virus and are suspected to play a major role in the transmission cycle. magpies (Pica pica). DVM. P. 2003 and 2004). S. Zeller.Biomathematics and Epidemiology Unit. France A STUDY ON FREE-RANGING WILD BIRDS TO BETTER UNDERSTAND THEIR ROLE IN THE CIRCULATION OF WEST NILE VIRUS IN CAMARGUE. PhD.J. we present preliminary results of serological investigations on different species of migratory passerine birds. Italy (1998). M. Kayser. SOUTHERN FRANCE E. West Nile (WN) virus is a flavivirus with a transmission cycle involving birds and mosquitoes. Russia (1999) and France (2000. National Reference Center for Arboviruses. Bicout. Seroprevalence was higher in several species of wild birds and experimental 60 . 1955). cattle egrets (Bubulcus ibis). outbreaks have been reported in humans or horses in numbers of countries including Romania (1996). Gauthier-Clerc. Murri. PhD . PhD. Anyhow. In this study. Station Biologique of La Tour du Valat. Jourdain. DVM. PhD student. In Europe. Institut Pasteur. While several bird species have been proven to be very sensitive to the NY strain of WN virus in America. the picture appears to be different in Europe. Sabatier. 1 INTRODUCTION Responsible for infections in humans and horses. Often asymptomatic. house sparrows (Passer domesticus) and tree sparrows (Passer montanus). In order to better understand and test different hypothetical roles of birds in WN virus circulation in Camargue. the infection with this flavivirus represents a threat to public and equine health. PhD KEYWORDS West Nile – Wild Birds – Serology – Camargue ABSTRACT West Nile (WN) disease is a mosquito-borne zoonosis transmitted in natural cycles between birds and ornithophilic mosquitoes. Recent outbreaks in the South of France enhance the need for further investigations on the role of wild birds in the ecology of WN virus in this area. The role of birds in WN epidemiology was first studied in Egypt in the 1950’s (WORK et al. Y. Indeed. it could be spread further by dispersing birds only at the end of summer when outbreaks most often occur in Europe. Although most human and equine cases are asymptomatic. Small passerine birds were captured with mist nests and were identified with respect to species. (3) tree (Passer montanus) and house (Passer domesticus) sparrows as they tend to disperse at the end of summer and are found numerously in the vicinity of horse farms and because they have been suspected to be a competent reservoir (KOMAR et al. we chose to concentrate on: (1) migrating passerine birds just after their arrival on the beach after crossing the Mediterranean Sea. several outbreaks occurred recently in horses and humans in different European countries including Romania (1996). They were all tagged with rings delivered by the National Museum of France. Blood was taken from the brachial vein 61 . including American crows (Corvus brachyrhynchos) (EIDSON et al. sex and age. we assumed that the virus was amplified by local birds mainly living in wet areas where mosquitoes are abundant. Our objective in this study was to assess the seroprevalence of WN virus in selected wild bird species captured in the Camargue area. the circulation of WN virus in bird populations is most often revealed by serological investigations only. As a consequence. The field work consisted of capturing the birds. Italy (1998). Russia (1999) and France (2000. And finally. In these two species. 2003) but no clinical sign was described in birds (MURGUE et al. it is important to study the circulation of this virus among birds in areas where recent outbreaks occurred. 2001). Since its introduction into North America in 1999. 2004). such as in Camargue in the year 2000 (MURGUE et al. WN virus has been responsible for the death of thousands of wild and captive birds and is still spreading north. 2002). (2) cattle egret (Bubulcus ibis) chicks. Given these hypotheses. Next. banding them in order to identify them individually and bleeding them. Magpies were caught with traps and cattle heron chicks were taken on the nest. because they lie exposed to mosquito bites in wet areas and the adults are often found in close contacts with horses. 2002). (4) magpies (Pica pica) as they belong to the Corvidae family which is well-known for being highly susceptible to the virus on the Western Continent (YAREMYCH et al. we made the hypothesis that WN virus was periodically introduced by migrating birds coming from endemic areas such as SubSaharan Africa. 2001).inoculations showed that sensitivity was variable depending on the species tested and was particularly high for house sparrows (Passer domesticus) and hooded crows (Corvus corone sardonicus). west and southward from its first outbreak area (New York City). 2 MATERIAL AND METHODS To select the species that we focused on. 2001). Mortality in birds was also described in Israel in white storks (Ciconia ciconia) (MALKINSON et al. WN virus can be responsible for encephalitis and even death in sensitive hosts. Nevertheless. viremia was high enough to infect mosquitoes. The spread of the disease is easily detected because of the high level of mortality observed in some species of birds. more than 500 hundred samples from migrating small passerine birds. from mid-June to mid-July for cattle egret chicks and from mid-August to the end of October for sparrows and magpies. The capture sites of sparrows and magpies are presented in Table I. 3 RESULTS At the end of the field work season. Heron chicks were sampled from different colonies. Montgomery. 196 from sparrows (144 from house sparrows and 52 from tree sparrows) and 34 from magpies were collected. Guilford. and the other one (site B) at the Marais du Vigueirat. Blood samples were screened for the presence of anti-flavivirus IgG with an appropriate indirect ELISA method. 220 from cattle egret chicks. Table 1: Capture sites of Tree sparrows. UK). Confidence intervals and other statistical results were calculated for a risk of error of 5% using the Binomial Law since we were out of the conditions of applicability of the Normal Law. TX) as described by EBEL et al. north-east of Aigues-Mortes. We used horseradish peroxydase-conjugated goat anti-chicken IgG (KPL Europe.into heparin capillary tubes. between Arles and Salin de Giraud. on the other side of the Rhône river. house sparrows and magpies Species Tree sparrows House sparrows Capture site Site A Site B Site A Site B Site A Magpies Site B Stes Maries de la Mer Other sites Number captured 37 15 60 84 23 5 4 2 34 Total captured 52 144 62 . Guilford. We also looked for ticks but only on the head in order to reduce as much as possible the time during which birds were handled. The sample season lasted from April to mid-May for migratory passerine birds. Magpies mainly came from site A but some of them were sampled from an outdoor cage where they were kept temporally in captivity by a trapper in the proximity of Les Saintes-Maries de la Mer. 2002). UK) and anti-wild bird IgG (Bethyl Laboratories. Inc. two other main sites were used: one (site A) at the Station Biologique of La Tour du Valat. anti-duck IgG (KPL Europe. Half of the sparrows were caught in site A (n=97) and half in site B (n=99). the main one being located at the Charnier-Scamandre.. (EBEL et al. The capture site of migrant birds in spring was located at the south of Salin de Giraud whereas at summer and autumn. 8% [0.3% [0-22].5] of the Passeriformes sampled were found positive for WN virus during the outbreak that occurred in Bucharest in 1996 (SAVAGE et al.0] 0. 1999) and 8% [4.3 [0.1-22. during WN epizootic in Camargue.3-27. one could have expected a higher prevalence in local birds such as sparrows and magpies.8 [3.2-38. In Poland.1] (HARS et al.4] We only found a few ticks: 23 on migratory passerine birds at spring and one on a tree sparrow in October.6] and 0% [0-3.4-99.4-28.2] of the tree sparrows tested between 1995 and 1996 were found positive to WN virus (JURICOVA et al.6] 0. The fourth one came from site A where seroprevalence can be estimated to 4.2] 11. where no recent outbreak had been described.0-3. 4 DISCUSSION Given the fact that an equine outbreak occurred during the same summer in the vicinity of Les Saintes Maries de la Mer. similar results had been obtained with a seroprevalence in magpies and sparrows estimated respectively to 22% [6. An unfed and unfixed adult male of the genus Rhipicephalus was also found on a European robin (Erithacus rubecula) at spring: this species of tick was actually very abundant on the capture site.4] of the house sparrows and 12.The commercial anti-wild bird conjugate proved to be efficient to detect antibodies from tree sparrows.0 [0.2] 75. If we assume that the captured sparrows belong to two different populations. Table 2: Sparrows and magpies serological results Species Capture site Site A Sparrows Site B All sites Site A Magpies Site B Stes Maries de la Mer Other sites All sites Number tested 97 99 196 23 5 4 2 34 Number positive 1 0 1 1 0 3 0 4 Seroprevalence (%) 1.9-6.5 [0. As a comparison.0 [0.0 [19.0-52.6% with a risk of error of 5% for the population at site B.0% [0-5. In the year 2000. 2. 63 . 1998). the results in Table 2 lead to conclude to a serological prevalence with WN virus of 1. 8% [0.0-5.4] two years after (GRANT et al.4-47.6] 0. The four positive magpies were hatch-year individuals with three of them coming from the area of Les Saintes Maries de la Mer. As shown in Table 2.0-84.0 [0.1% [3. 2004). the positive sparrow (of undetermined age) was captured in site A.6] for the sparrow population at site A and a prevalence smaller than 3.0 [0. house sparrows and magpies.15-13.8] 4. 2001).0-2. Most of them were nymphs of the genus Ixodes and three were Ixodes larvae.4] 0. if they are carried by migrating birds. We found few hard ticks on migratory passerine birds and it will be interesting to check whether the birds from which ticks have been removed are positive in WN serology. 2003). Even if WN virus was circulating among birds populations earlier in the season. most of them were nymphs of the tick species Ixodes ricinus and experimental infections have suggested that this species of tick does not support replication of WN virus (LAWRIE et al. and in the survival of the virus over winter. magpies and sparrows could be two species involved in the spread WN virus at a regional scale. the hypothesis of a new arrival of WN virus because of migratory birds will be reinforced as well. 64 . To conclude. specially for young birds after leaving the nest. In the same way. Ticks have been suspected to play a role in the introduction of WN virus. this will be in favour of the hypothesis that the virus was amplified in those birds before being responsible for clinical disease in horses. Indeed. making them a potential reservoir and candidate for WN virus translocation. it was probably at a very low level. This is nevertheless essential in order to understand the epidemiological cycle of diseases such as WN disease in non domestic species. magpies are also good candidates that could multiply and spread WN virus. As a consequence.Results of the serological investigations on wild birds are consistent with those from sentinel birds. if we find antibodies in samples from migratory birds coming from Africa. Whereas ducks and domestic poultry were shown to have low viremia. As they belong to the Corvidae family. if we manage to find antibodies in blood samples from cattle heron chicks. Actually. when one would have expected it to be higher. our results seem to indicate that ticks are not really frequent in summer and autumn in sparrows and magpies. sparrows are known for having high levels of virus in their blood in experimental infections (KOMAR et al. it is probable that we will not find antibodies against WN virus in the species sampled at spring. Nevertheless. Even if we probably underestimated the real parasitic burden because small ticks such as larvae are difficult to see. Our results indicate that WN virus circulated in those two species at the end of summer 2004. Because of their dispersing behaviour. The results obtained in turn are often disappointing in comparison with the involvement. 2004). we would like to underline the fact that field work with free-ranging wild birds requires a huge investment of time and necessitates the participation of several qualified field workers. Both studies indicate that WN virus was circulating among birds at the time of the epizootic in horses. seroprevalence was low in birds captured during the outbreak in horses. That is why it is necessary to develop and adapt laboratory methods of analysis to make it possible to work on samples from wild species. Emerg Infect Dis 2003. AUGÉ P and CHAVERNAC D. AUTHORS ADRESS E.fr 65 . 42: 182 .322. 951: 94 . Introduction of West Nile virus in the Middle East by migrating white storks. DUPUIS A P and NICHOLAS D. PANELLA N A and BURNS J E. Ecole Vétérinaire de Lyon. 2. JURICOVA Z. Surveillance de l‘infection de l‘avifaune camarguaise par le virus West Nile. We also want to thank the Station Biologique of La Tour du Valat and the Marais du Vigueirat for allowing the capture of wild birds on their land. Epidemic West Nile encephalitis in Romania: waiting for history to repeat itself. West Nile outbreak in horses in southern France. Avian Dis 1998. montanus) in Poland. 7: 692 . 8. 8: 392 . 5 CITATION INDEX 1. GRANT L. France Email: e. Jourdain.397. Emerg Infect Dis 2001. BANET C and WEISMAN Y. and bunyavirus arboviruses in house sparrows (Passer domesticus) and tree sparrows (P. 3. EIDSON M. 8: 979 . 4. LANGEVIN S and HINTEN S. 7: 615 . 9: 311 . 7. Faune sauvage 2004. Experimental infection of North American birds with the New York 1999 strain of West Nile virus.657. MURRI S and ZIENTARA S. 10: 653 . Emerg Infect Dis 2002.620. Emerg Infect Dis 2001.ACKNOWLEDGEMENTS We want to thank the Parasitology Unit of the Veterinary University of Lyon for their help in the species identification of the ticks removed from birds. Ixodid and Argasid tick species and west nile virus.185. Serologic Evidence for West Nile Virus Infection in Birds in the New York City Vicinity During an Outbreak in 1999. MALKINSON M. 7: 621 . Emerg Infect Dis 2002.101. 2000: the return after 35 years. 9. DVM. Emerg Infect Dis 2004. Antibodies to alphavirus.jourdain@vet-lyon. Emerg Infect Dis 2001. 1 avenue Bourgelat.625. Ann N Y Acad Sci 2001. CAMPBELL G L and CORNELIA S. 6. flavivirus. 1999.696. HARS J. 5. PhD student Unité de Biomathématiques & Epidémiologie. The rest of references are available from the author. Detection by enzyme-linked immunosorbent assay of antibodies to West Nile virus in birds.982. PINOWSKI J and LITERAK I. 69280 Marcy l’Etoile.58. EBEL G D. MURGUE B. LAWRIE C H. KOMAR N. KOMAR N. 10. Crow deaths as a sentinel surveillance system for West Nile virus in the northeastern United States. et al. 261: 54 . KOMAR N and SORHAGE F. 2003) O. KEYWORDS Raptors – Waterbirds – Exposure – Anticoagulant rodenticides – France ABSTRACT This presentation is a synthesis of two epidemiological surveys performed in Loire Atlantique (France. exposure of raptors (Falco tinnunculus. bromadiolone. B. exposure of non migratory raptors (Falco tinnunculus. Buteo buteo. coumatetralyl and difenacoum) was compared. they are used against rats. especially for predators and scavengers (BERNY et al 1997. 44) on carcasses of birds (raptors and waterbirds) collected in the wildlife center of the national veterinary school of Nantes: birds were either dead when they were brought to the center or died within a week. M L’Hostis. The aim of these studies was to evaluate their possible exposure to anticoagulant rodenticides: In 2002. In 2003. Tyto alba and Strix aluco) and waterbirds to five anticoagulants (brodifacoum. National Veterinary School of Nantes. France WILD BIRDS EXPOSURE TO ANTICOAGULANT RODENTICIDES IN LOIRE-ATLANTIQUE (FRANCE).. STONE et al 66 . Pouliquen.Wildlife Center. mice and particularly vole (Arvicola terrestris) and coypu (Myocastor coypus). In France. De la Cotte. N. Buteo buteo. bromadiolone is the most frequently rodenticide detected or quantified in liver of birds. Lambert. Tyto alba and Asio otus) to two anticoagulants (chlorophacinone and bromadiolone) was studied. apples or cereals. carrots. They are presented as baits. A 2 YEARS STUDY ON WATERBIRDS AND RAPTORS (2002 . Philippe. 1 INTRODUCTION Anticoagulant rodenticides are traditionally used to control rodents worldwide. The anticoagulant rodenticides exposure is higher for raptors than for waterbirds. Many incidents of non-target wildlife mortality from anticoagulant rodenticides occurred. H. coumafen. 3 RESULTS 3. Extracts were purified by a solid-phase extraction technique using C18 reversedphase cartridges. The purpose of the studies was to evaluate the exposure to such products of wildbirds (raptors and waterbirds). One kestrel falcon had chlorophacinone between limits of detection and quantification (≥ 0. 9 barn-owls (Tyto alba) and 10 long-eared owls (Asio otus). Birds were analysed for five rodenticides: difenacoum. 13 (26%) had rodenticides in liver: 5 with chlorophacinone. Analyses for anticoagulants were completed at the Toxicology Laboratory of the National Veterinary School in Nantes. Anticoagulant compounds were extracted from liver with mixture of organic solvents (acetone.5 µg/g).04 et ≤ 0.4 kestrel falcons (Falco tinnunculus). 13 common buzzards (Buteo buteo). Birds were anaylsed for two rodenticides: chlorophacinone and bromadiolone. coumatétralyl.4 µg/g : 3 kestrel falcons (n=19). Purified extract were then injected onto a HPLC C18 end-capped reversed-phase column. Ten showed bromadiolone and chlorophacinone concentrations in liver above 0. Anticoagulant compounds were identified and quantified by using fluorimetric detector set at an excitation of 318 nm and an emission wavelength of 390 nm. diethyl ether and chloroform). brodifacoum. 67 . The liver was removed from each carcass and stored at -18°C until analyses. Among the 51 raptors.1999). 11 common buzzards (Buteo buteo) – 15 nocturnal raptors – 10 barn-owls (Tyto alba). 5 tawny owls(Strix aluco) – and 28 waterbirds – 15 mallards (Anas platyrhynchos).4 and 2.11 et ≤ 0. bromadiolone. First study (2002). 13 black coots (Fulica atra) and 1common moorhen (Gallinula chloropus). 51 carcasses were used: 19 kestrel falcons (Falco tinnunculus).1 First study (2002). Complete necropsies were performed on birds immediately (due to the rapid degradation of anticoagulants) after their death either at their reception or during their hospitalization (within one week). Second study (2003).33 µg/g) and bromadiolone was detected in one common buzzard between limits of detection and quantification (≥ 0. coumafene. 2 MATERIALS AND METHODS Birds of these surveys were collected by private individuals or wildlife protection associations because they were affected and were brought to the Wildlife Center of the National Veterinary School in Nantes for hospitalization. 58 carcasses were used : 15 diurnal raptors .12 µg/g). 2 common buzzards (n=13). 1 long-eared owl (n=10) and 4 barn owls (n=9). 7 with bromadiolone and one long-eared owl had both bromadiolone and chlorophacinone in detectable concentrations (respectively 1. For all the 26 contamined birds. 4 DISCUSSION Our results show that the raptors exposure to anticoagulant rodenticides is relatively large and similar findings were reported by STONE et al. 1996 . BYERS RE and KAUKEINEN DE. a 4-year survey. 1997. 2003) of the incidents of poisoning by anticoagulants in New York respectively from 1971 to 1997 and from 1998 to 2001. 35: 1817 . et al. 48: 212 . 2. difenacoum in 8 birds. J Wildl Manage 1984. 5 CITATION INDEX 1. The 3 most common anticoagulant rodenticides detected or quantified in the liver are respectively bromadiolone. which are more susceptible to eat official baits. difenacoum and chlorophacinone) were most frequently detected in non target wildlife in Europe (SHORE et al. At least one anticoagulant rodenticide was detected in the liver of 15 birds or quantified in the liver of 11 birds. BERNY P.3. However we can’t determine the effects of such exposure on their health. FREESTONE P and KITCHENER AC. SHORE RF. 1996. Chemosphere 1997. BURONFOSSE F. its concentration was low (<1 µg/g) except one common buzzard (coumafene liver concentration = 2 g/g) and one black coot (coumafene liver concentration = 23.216. 1999). In contrast brodifacoum is implicated in 80% (STONE et al. brodifacoum appears to have the greatest potential for non target wildlife mortality. coumafene in 5 birds. difenacoum and chlorophacinone. 1997).2 Second study (2003). polecats) are commonly contaminated by anticoagulant rodenticides (SHORE et al. 1999) and 84% (STONE et al. In Loire Atlantique and especially in wetlands. BERNY et al.282. bromadiolone is widely used as carrots or apples against coypu (Myocastor coypus). Envir Pollut 1996. 91: 279 . Bromadiolone was detected or quantified in 15 birds. When anticoagulant rodenticide was quantified in the liver. the low exposure of waterbirds. STONE et al. Field evidence of secondary poisoning of foxes (Vulpes vulpes) and buzzards (Buteo buteo) by bromadiolone. 6 (only raptors) were contamined by 2 or 3 different anticoagulant rodenticides. BURONFOSSE T. (2003). 3. 1984). such as raptors or carnivorous mammals (foxes. Secondgeneration rodenticides and polecats (Mustela putorius) in Britain. Other prior studies showed that these rodenticides (bromadiolone. Direct exposure of birds like raptors after ingestion of traited baits appears unlikely because of their feeding diet and the exposure seems to occur by the consumption of contaminated rodents (MERSON et al.1828. According to our second study. 68 . brodifacoum in 4 birds and coumatetralyl in only one bird.52 µg/g). MERSON MH. BIRKS JDS. Anticoagulant rodenticides were detected or quantified for 73% (n=22) of all raptors (n=30) and 14% (n=4) of all waterbirds (n=28). Residues of the rodenticide brodifacoum in voles and raptors after orchard treatment. BERNY et al. that is confirmed by our results. Prior surveys also indicated that predators. confirms a secondary exposure for raptors. STONE WB. 5. 70: 34 . 187 .193.armauny@free. France Email: cds. 44307 Nantes cedex 03. OKONIEWSKI JC and STEDELIN JR. Anticoagulant rodenticides and raptors: recent findings from New York.4. Poisoning of wildlife with anticoagulant rodenticides in New York. Ecole Nationale Vétérinaire de Nantes. Bull Environ Contam Toxicol 2003. OKONIEWSKI JC and STEDELIN JR. J Wildl Dis 1999. 1998-2001. Lambert Centre de Soins de la Faune Sauvage. STONE WB.40. AUTHORS ADRESS O. Atlanpole – La Chantrerie.fr 69 . poor response to therapy. Of particular concern for the white winged wood duck is a presumed immune system deficiency due to poor genetic diversity. lack of early symptoms. 1992). mycobacterial infections can be devastating to captive waterfowl collections. previously 70 . Improved exhibit and holding design. Ohio. and endoscopy has been attempted to help manage waterfowl mycobacterial infections but as of yet. resulting in over 80% mortality in some collections.Akron Zoological Park. The WWWD population. United States of America MYCOBACTERIAL INFECTION IN WATERFOWL COLLECTIONS : A CONSERVATION PERSPECTIVE. make disease prevention particularly difficult. utilization of ultraviolet lighting. and altering soil pH (above 7) are being examined as possible tools to minimize organism build-up and thereby control the morbidity and mortality of exposed waterfowl.UV light – pH – Band . Riggs DVM. ELISA. Ante mortem mycobacterial screening involving PCR. CBC. resulting from limited population founders of unknown genetic relationship. ABVP (Avian) KEYWORDS Waterfowl – Mycobacteria – tuberculocidal – SSP . There are currently approximately 60 WWWD individuals in captivity in the SSP managed program. Poor genetic diversity within species (ducks and others) has been implicated as contributing to immune system failures to resist mycobacterial infections (CROMIE et al. 2001). The ubiquitous nature of the mycobacterial organisms.White winged wood duck . G. Of particular concern is the captive management of endangered species. optimization of husbandry. and immune system deficiencies (due to genetics or prior exposure). such as the white winged wood duck (WWWD) which represent the only anseriforme Species Survival Plan (SSP) of the American Zoo and Aquarium Association. poor performance of disinfectants in the field. A contributing factor to their low numbers in captivity is their historical predilection for mycobacterial disease. Bird and Exotic Specialty Hospital. has not proven sensitive or specific enough for adequate disease control (TELL et al.Sharing coefficient ABSTRACT Although relatively rare in wild waterfowl (FRIEND 1999). Reproductive stresses. 71 . WWWD’s in their natural habitat have limited exposure to mycobacteria and. overcrowding. Mycobacterial infections are relatively rare in wild waterfowl. avium serotypes 1. with a 0. poor ventilation. Dabbling ducks are also more likely to be infected than grazers since they feed in the favoured growth environment of the mycobacterial organisms (CROMIE et al. All waterfowl are potentially susceptible to mycobacterial infection although ducks and swans are more represented than geese. substandard nutrition. and management decisions (e.6% incidence rate in total wild bird populations (FRIEND 1999).5% genetically diverse per the SSP studbook in 1992.3% incidence rate. There are no pathognomonic signs. are likely to have little “historical” immune system experience with the organisms. enclosure. Recently. but can also reflect the severe impact mycobacteria can have on waterfowl populations. compared to 3. 2. This evolving trend further complicates ante mortem diagnosis by requiring testing modalities to be sensitive to both M. and hopefully by improving genetic diversity in the future. and quarantine protocols in the short term. Mycobacterial infections are characterized as chronic. Waterfowl mycobacterial infections are generally caused by organisms of the Mycobacterium intracellulare complex. in some surveys up to 70% of isolates. This alludes to the fact that the organisms thrive in the shallow waters of the soil water interface. and 3 (TELL et al. 1991) and to be 63. has instead been shown by our study to have diversity no better than first generation relatives (USA population). possibly contributing to their continuing sensitivity to mycobacterial infections in captivity.g. 1992). Incidence in captive collections can be much higher. therefore. 1 INTRODUCTION Mycobacterial organisms have been referred to as “ducks of the microbe world” (GRANGE 1987). symptoms relate to target organ and degree of damage. pinioning) can greatly exacerbate the disease process (CROMIE et al. 2001).presumed to have 81% heterozygosity in a 1989 report (TOMLINSON et al. genovense in order to be effective screening tools. most commonly by M. 1991). 2001). avium and M. As with most diseases. debilitating diseases usually seen in individual birds rather than in epizootics. It is our feeling that the presumed “increased sensitivity” to mycobacteria of this population can be controlled with improved husbandry. stress can play a major role in determining which individuals within an exposed population will become chronically infected with mycobacteria. They are characterized by numerous caseous lesions in the GI tract and major organs. due in large part to exposure variance. an increasing number of mycobacterial isolates have been determined to be Mycobacterium genovense (TELL et al. Captive holding techniques commonly employed have encouraged infection by maximizing both stress on the captive individuals and exposure to mycobacterial organisms. low level environmental exposure by young waterfowl has been found to decrease resistance to pathologic mycobacterial later in life (CROMIE et al. although this is frequently unreliable early in the disease and can drop off late in the course of the disease. corresponding to immune system collapse. 1991). long held as the test of choice. Elevated white blood cell counts are often reported with marked monocyte shifts. and in nearly 100% of municipal water supplies in some studies (FALKINHAM 2003). Mycobacteria. in particular M. coli). avium. they are difficult to physically wash off of contact surfaces. “Tuberculocidal” refers to organism destruction on non-porous surfaces with extended contact times at room temperatures (CARSON 1978). They are normal soil inhabitants and can persist protected underground and multiply in earthworm and amoeba hosts. are notoriously resistant to most common disinfectants in the field (FALKINHAM 2003). They thrive in a variety of environmental conditions and can persist even in seasonal sub-zero temperatures for up to seven years (THOEN and RICHARDS 1977). an ELISA did prove predictive in the mid to later stages of disease in selected duck populations. due to their adherent properties. Serology has also been historically unrewarding. Currently no “gold standard” test exists. and lower temperatures will inactivate the disinfectant or require a much longer contact time than is feasible in the field.Early diagnosis of this relatively common disease can prove difficult. endoscopy and biopsy are utilized by some institutions for mycobacterial screening. Acid fast faecal staining. 72 . is at best positive in only 10%-30% of positive avian mycobacterial cases. Hot water is also not the answer. Finally. 2000). Mycobacteria (both pathogenic and non-pathogenic types) are ubiquitous organisms found routinely in most environments. In addition. Natural surfaces are seldom non-porous and organic material. With the inherent diagnostic and treatment difficulties. Even disinfectants labelled as “tuberculocidal” cannot be depended on for field use. waterfowl can also shed huge numbers of organisms into the environment for extended periods of time prior to displaying demonstrable clinical symptoms. Even this is limited in its absolute ability to declare birds negative and presents time/ risk/cost implications. Environmental disinfection is unfortunately not a viable solution to the problem. and are resistant to ozone and most common safe disinfectants. although in one study (CROMIE et al. They have been cultured in up to 40% of surface waters. Mycobacteria have routinely been shown to grow in chlorinated water (500 times as resistant as E. This sub-clinical environmental contamination enhances and easily perpetuates the epizootic conditions in many waterfowl collections. hard water. prevention and control are the mainstays of collection protection. Once infected. Mycobacterial cultures are relatively specific but do not often show high sensitivity and require long incubation times for results. Radiology can be a beneficial diagnostic tool (Bush et al 1978) but is effective in only the 10% of cases with bone lesions. Mycobacteria thrive in hot water and will even grow in hospital hot water heaters. The problem is made more difficult by the prevalence and resistant nature of the organism(s) involved. Compounding the problem. as poor genetics and exposure factors have played a significant role in the problem. They are listed CITES I and represent the only anseriforme SSP maintained by the American Zoo and Aquarium Association.1.5%. UV light is only effective. This was not a controlled study and obviously more work is needed to determine successes and limitations. To be successful at minimizing mycobacterial buildup. Shaded areas or areas where organisms are shielded by soil will not be affected by UV radiation. WWWD’s have had severe mycobacterial problems in captivity. At WWT. does show potential as a mycobacterial deterrent. however. Our current study unfortunately has shown it to be much worse. disease surveys found a significantly higher mycobacterial infection rate in shaded pens as compared to pens directly exposed to sunlight (CROMIE et al 1992). 1. breeding recommendations are in place for all institutions holding pairs and it is hoped to build numbers back up to stable levels within the near future. down from the 120 birds previously maintained by the SSP. A Johne’s study (M. but this technique seems to hold promise to help minimize environmental build-up in exhibits. with direct mycobacterial contact. and Java. Human hospital studies have demonstrated complete mycobacterial disinfection in water suspensions at 15 mW-s/cm2 (Manning unpublished). in pairs. The white-winged wood duck story White-winged wood ducks are large. Approximately 60 individuals currently exist in the USA’s captive programme. Currently.Maintaining an alkaline soil. or small groups that feed nocturnally in shallow still waters. 73 . however. tree perching. the numbers in the wild are critical. and eliminate the shading of soil-water interface areas. paratuberculosis) resulted in a ten-fold decrease in paratuberculosis cases after lime treatment of pasture soil increased the soil pH to over 7 (COLLINS 2001). Sumatra. Research in other species has shown that this lack of genetic diversity can have severe impact on the immune status of the individual. predisposing to mycobacterial infections. Thailand. the genetic diversity within the SSP was felt to be only 63. they live solitarily. Our data has determined the DNA relatedness of the current breeding SSP population to be no better than first order relatives. Mycobacterial disease has played a significant role in the numbers decline in captivity. Many hospital ventilation sanitation systems utilize 100 mW-s/cm2 UVC devices for total mycobacterial airborne sanitation. Known in some areas as the “Spirit duck” for their haunting vocalizations. Due to the limited number of founders of the captive WWWD population (7). Due primarily to habitat pressures. UV light may also be an effective anti-mycobacteriocidal agent for captive collections. deep forest ducks native to Assam. enclosure design must incorporate the ability to minimize persistent shading (or rotate shaded areas). 1992) have been linked to avian mycobacteriosis. We feel these high rates in captivity may not accurately reflect the true sensitivity of the wild species. Up to 84% of individual deaths in certain collections (CROMIE et al. samples were taken for DNA analysis from the pilot group (31 individuals) serving as the source for most individuals currently held in the US SSP. The introduction of the wild caught naïve founder population into a large mixed collection endemic with mycobacterial has had tragic results. Their method of dabble feeding at the soil-water interface in shaded areas places them in contact with the highest mycobacterial concentrations in captivity. Standard bird species assays showed little or no genetic variability within the population. A solitary. These ducks represent nearly all of captive breeding stock in the United States and are direct descendants from the original captive breeding stock in the UK. The average band-sharing coefficient 74 . Therion International Labs performed the testing of the collected samples. Not only did it lead to large numbers of individual losses. 2 MATERIALS AND METHODS To investigate a genetic component contributing to the high infection rate of the species. Arguably. the WWWD SSP has undertaken several pilot projects in an attempt to better understand and improve the mycobacterial diagnostic testing (PCR. is now most frequently kept as a pinioned. Quarantine and necropsy protocols have been developed to investigate and screen for current infections and pilot projects are underway to determine genetic diversity and assess new ante mortem avian mycobacterial screening techniques. Digested fragments were separated by size with gel electrophoresis. it led to losses in an already small irreplaceable genetic pool. where mycobacterial contact is minimal. Currently. The resultant increase in stress on the individual would further minimize resistance to the mycobacterial organisms it now routinely contacts. DNA was isolated using non-organic extraction. ELISA) and genetic status of white-winged wood ducks in captivity. Furthermore. infected individuals may well have been unknowingly disbursed to other collections to perpetuate the mycobacterial cycle. tree perching duck in the wild. DNA samples were cleaved with restriction enzyme Alu I. but more importantly. due to endemic mycobacterial levels at the initial captive propagation site. ground bird in large mixed groups. Two additional assays were undertaken that yielded 72 genetic markers with an average of 45 markers per individual. 3 RESULTS DNA profile analysis was performed on 31 white winged wood ducks to estimate relative relatedness of the birds in the captive conservation program. all known captive WWWD’s descend from the twelve individuals (TOMLINSON et al 1991) brought into captivity at the Waterfowl Trust in the late 1960’s.Husbandry failures have also played a major role in the WWWD’s mycobacterial problems. then transferred to a nylon membrane and hybridized sequentially with radioactivelylabelled multi-locus DNA probes OPT-02 and OPT-05. Natural husbandry traits also work against WWWD’s in captivity. Minimizing soil-water interface on ponds. If we are to assume that the genetic inbreeding has also contributed to immune system deficiencies that predispose the population to mycobacterial infections. specific pairings.80 and 0. Maintaining birds in breeding pairs or small non-breeding groups. Results of this study displayed a very low genetic diversity within the population. Minimizing temperature extremes (50-80 degrees F). Maximizing nutrition. 75 . 7). It is imperative that genetic diversity of the captive species be improved within the foreseeable future if we are to truly function as a “species survival plan”. 2). Within the tested population.41 with only 2-2. 3). Minimizing shaded areas and allow rotation of shaded areas for periodic UV exposure where possible. Our immediate goal is now to analyze any potential non-related individuals in collections outside of the United States to determine if significant improvements in breeding can be accomplished with currently held individuals.00. Current white-winged wood duck SSP husbandry recommendations include: 1). it increases the importance of vigorously pursuing husbandry protocols and diseased bird culling to minimize environmental build-up of the mycobacterial organisms.31-0. Husbandry recommendations to more closely mirror natural behaviour will be put into place via the SSP and we will continue to evaluate the efficacy of UV light and soil pH changes in the control of environmental mycobacterial contamination. 8).65-1. 4). we will continue our pilot projects to submit samples for the development of a sensitive and specific ante mortem diagnostic screening tests. 5). The WWWD population’s genetic variability appears no better than would be expected from first-order relatives. To facilitate these goals. estimated heterozygosity were 0. We will also implement faecal PCR screening into SSP quarantine protocols and continue aggressive necropsy screening to attempt to analyze collection contamination from new acquisitions.77 for combined assays. Constructing ponds to be cleanable with moving water. 4 DISCUSSION The results of the genetic analysis have major implications on the SSP’s management of the species in captivity. Routine health screening/aggressive work up on “wasting” and ill individuals. other than for minor phenotypic variations.3 alleles per locus.75 for the respective assays and 0.(BSC) per pair was 0. BSC values can be ranked as breeding guides to maximize genetic variability within a population. 6). The range of BSC for the tested individuals was 0. are not critical and would not be expected to improve genetic variability within the captive population. Maintaining non-pinioned birds where possible. much lower than was previously thought. Since no individual is farther apart than a first order relative. 8. Mycobacteria isolated from exotic animals. Wildfowl 1991. of Interior 1999. Vol 9: No. (ed): Mycobacterial infections of zoo animals. 4. 5 CITATION INDEX 1. 4873 Richland Ave. Cairina Scutulata. CROMIE R. Smithsonian Institution Press 1978. University of Wisconsin. Appl Environ Microbiol 1978. J Am Vet Med Assoc 1977. CDC Emerging Infectious Diseases 2003. Encourage free flight exhibits with natural perching/nest boxes. In: Friend M. AUTHORS ADDRESS Gary Riggs DVM. Bird and Exotic Specialty Hospital. and new genetic lines can be introduced into the SSP population. CROMIE R. 6. Franson C (eds): Field Manual of Wildlife Diseases. we look forward to simplifying husbandry guidelines and minimizing the impact of individual disease entities on the captive population of this important species. TOMLINSON C. 24: 169 . Improving the management of a highly inbred specie: the case of the White-winged wood duck Cairina scutulata in captivity. RICHARDS W. Ohio 44203. Survival. THOEN C. It is anticipated that achievable husbandry and genetic improvements can make demonstrable changes in the current and future captive SSP white-winged wood duck population. 170: 987 . Slimbridge Center (1976-1991).5. 10.990. In: MONTALI R. 11).98. at the Wildfowl and Wetlands Trust.9). COLLINS M. FALKINHAM J. 43: 211 . Growth characteristics of atypical mycobacteria in water and their comparative resistance to disinfectants. CROMIE R.204. Tuberculosis.185. MACE G.214.846. 72: 105 . Avian immune response to mycobacterium avian: the wildfowl example. BUSH M. ABVP (Avian) Akron Zoological Park. Dev and Comp Immun 2000. 9. Minimizing shaded soil areas and maintain soil pH at 7 or greater. School of Vet Med. 42: 123 . Exclude wild fly-ins.1 . CARSON L. 36: 839 . 2. MONTALI R.109. Johne’s Information Center. Clinical experience with tuberculosis in exotic birds. Susceptibility of captive wildfowl to avian TB: the importance of genetic and environmental factors. 93 .133. 3. US Dept. 5. 7. Mycobacterial aerosols and respiratory disease.com 76 . FRIEND M. 2001. 199 . United States of America Email: garyriggs@aol. Wildfowl 1992. 10). Norton. As improved ante mortem testing develops. ROFFE T. The epidemiology of avian tuberculosis in White-winged wood ducks. Tubercle 1991. 7. 1996. peregrine falcons (Falco peregrinus) (DÖTLINGER and BIRD 1995. 1. had been studied to compare them with the purebred species in the literature and to establish reference parameters for those hybrid species. The haematological blood picture can give significant hints for the avian practitioner about e.Gyr-peregrine hybrid falcon . R. In the United Arab Emirates. dehydration. those falcons are seen in an increasing number in the Abu Dhabi Falcon Hospital. anemia. INTRODUCTION Although the saker falcon (Falco cherrug) is the traditional hunting bird in the MiddleEastern Countries. JENNINGS 1996) and gyr falcons (Falco rusticolus) (WERNERY et al. 2004).Haematology . Mannil KEYWORDS Birds . George and A. Muller. Although the haematological parameters are wellknown in saker falcons (Falco cherrug) (SAMOUR et al. infections and aspergillosis (CAMPBELL 1988). gyr-saker hybrid falcons and gyr-peregrine hybrids falcons. peregrine falcons and saker falcons are widely discussed in the literature. JENNINGS 1996).g. Abu Dhabi. T. only little information is available about those two hybrid species (WERNERY et al. United Arab Emirates HAEMATOLOGICAL VALUES OF GYR HYBRID FALCONS M. A. United Arab Emirates. One of the main reasons is the new law Cites of October 2002 allowing only captive-bred falcons to be used for falconry purposes inside UAE.Abu Dhabi Falcon Hospital. Due to the captive breeding of hybrid falcons. The haematological parameters of two different hybrid species. G. Due to the importance of the haematological blood parameters for the early detection of diseases in falcons.Gyr-saker hybrid falcon . 2004). 77 .Values ABSTRACT Haematological values for falcon species like gyr falcons. the number of captive-bred gyrsaker hybrid falcons and gyr-peregrine hybrids falcons used for falconry has rapidly increased over the past few years. this study covers the haematological parameters of complete blood counts (CBC) for gyr-saker and gyr-peregrine hybrid falcons. This leads to a change in the patients frequenting falcon clinics and hospitals throughout UAE as well to a great need to enhance research on these hybrids for the sake of their health. it is not the commonly used falcon for falconry in the United Arab Emirates anymore. 2. the cells of 5x16 squares have been counted in the center of the counting grid. In our study. After 5 minutes waiting time.0 ml EDTA tube (Teklab). This diluted sample has been filled via a capillary tube in the improved Neubauer haemocytometer. 0. The blood count was performed directly after blood taking with a maximum delay time of 30 minutes. UK). 31.9ml of a 1% ammonium oxalate solution had been mixed with 100µl of the falcon blood sample and kept on a tube roller for 3 minutes. After 5 minutes waiting time.3g trisodium citrate and 1000ml distilled water. All falcons that were showing clinical signs or that were diagnosed with disease were excluded from in this study. A small amount of the diluted samples has been filled via a capillary tube in the improved Neubauer haemocytometer. the cells of 4 outer large squares have been counted in the center of the counting grid. then: N = RBC x 1012 / l 100 White blood cell count (WBC) (SAMOUR et al 1996) 1. then: N = WBC x 109 / l 20 Packed cell volume (PCV)/ Haematocrit (Hct) (SAMOUR et al 1996) The PCV is measured by the used of microhaematocrit capillary tubes (Fortuna. 2.1 Material The haematological parameters of 689 clinically healthy hybrid falcons were evaluated. The counted cells had been calculated as follows: N=Number of cells counted. the haematocrit is used as this might be the more accurate than the PCV (FUDGE. After taking blood from the right or left basilic vein (Vena cutanea ulnaris superficialis) or the right or left caudal tibial vein (Vena metatarsalis plantaris superficialis). 2000) Mean cell volume: PCV x 10 = MCV(fl) RBC Mean cell haemoglobin: Hb x 10 = MCH (pg) RBC Mean cell haemoglobin concentration: Hb x 100 = MCHC (g/l) PCV 78 .5ml blood was stored in a 1. Four ml of this solution has been mixed in a plain sample tube with 20µl of the falcon blood stored in the EDTA tube. Germany) and centrifuge (Hawsley. MATERIAL AND METHODS 2. The counted cells had been calculated as follows: N=Number of cells counted.2 Method Red blood cell count (RBC) (SAMOUR et al 1996) The working solution consists of 10ml 40% formaldehyde. The complete blood count (CBC) of 369 gyr-saker hybrid falcons and 320 gyr-peregrine hybrid falcons has been examined manually. 40±0. The excess solution is allowed to drain.99 5.29-243.33±0.15 range 2.88 44.22 49.66 221.72 46.83 34.24±6. 3.57 7. then allowed to dry and examined under oil immersion lens. The optical eye of the HemoCue microcuvette contains reagents deposited on its inner wall and the blood sample is drawn in to the cavity by capillary action spontaneously mixed with the reagents. The slide is dipped in fixative solution for 5 seconds.18-2.92 0.07 32.1 for 5 seconds and than in solution 11 for 5 seconds.23-19.02-53. Results The evaluation of the 369 (Falco rusticolus x Falco cherrug) hybrid falcons showed the following picture: Table 1: Haematological parameters for gyr-saker hybrid falcons (Falco rusticolus x Falco cherrug) Parameters RBC (x1012/l) HB (g/dl) Hct % MCV (fl) MCH (pg) MCHC (g/dl) WBC (x 109 /l) Heterophils % Lymphocytes % Monocytes % Eosinophils % Basophils % n=369 2.85±21. Staining method The HEMAstain test is for the rapid.57±4.29±0.28-9.90±3.11±2.37-2.78 41.23 46.21 0.58 1.85-35.64 76. but is not part of this study.02 2.15 17.41-83. Position a spreader slide in front of the drop of blood at an angle of about 45° and move the spreader backwards and touch gently the drop of blood which will run across the edge of the slide. Preparation of blood smear Take one small drop of blood sample in a clean microscopic slide.Haemoglobin The haemoglobin is measured by capillary. than in solution. Push the spreader with a steady forward movement to create a blood smear.23 200.42±1.50 51. The slide is rinsed with distilled or deionised water.25-0. the system provides a direct reading of the concentration of haemoglobin in a blood sample.50±2.91-56.73±1.42±1.00 0.20-47.84-6.48 16. differential staining of haematological smears that yields qualitative results similar to Wright-Giemsa stain.91 4. venous or arterial blood used in EDTA. Used with the HemoCue photometer.55 79 .49 69. The examination of the blood smear for the shape of the blood cells and avian blood parasites is routinely performed for each blood smear. 18±0.00-85.00-33.04 32.50 ---14.06 138.75 219.26 17.95±12.56±3.50±0.29-2.81 6.41 40.01 32.39±0.00±4.27 49.30 2. Table 3.91 5.03 0.65 16.59±8.014 range 2.94 37-53 11.08 0.44±0.60 46.06 0.43±1. lymphocytes.90-35.06 45.47 47.50 44.32±3.32-245.60 0-0.00-8. DISCUSSION Extensive work has been done on the haematological parameters of peregrine falcons.91±3.80 ----188-146 40.55±2.28-9.40±0.00-48.18±3. Although the range of some parameters is relatively close.10 ---9.33-19.43±6.00 1.The 320 evaluated (Falco rusticolus x Falco peregrinus) led to those results: Table 2: Table 1: Haematological parameters for gyr-peregrine hybrid falcons (Falco rusticolus x Falco peregrinus) Parameters RBC (x1012/l) HB (g/dl) Hct % MCV (fl) MCH (pg) MCHC (g/dl) WBC (x 10 /l) 9 n=320 2.60 2.25-56.7±25. WBC.26 80 .20±12.45 34.65 0.21 ---14.77-6.37±1.6 DOETLINGER and BIRD 1995 n=48 3.51±7.40±1.70 --------12.20 3.20±13.14-84.54 2.49±0. significant differences can be seen in MCV.30±0.75 194.08 67.90 ---- WERNERY et al 2004 n=138 3.32±4.82±1.63 1.48 7.38 75.14±0.38 117.36 4.80-18.67±2.45 0-0 Heterophils % Lymphocytes % Monocytes % Eosinophils % Basophils % 4.3 0-0.82-47.00 14.57 52.52 0. heterophils.00 55.9±1. Literature comparison of haematological parameters in peregrine falcons (Falco peregrinus) Parameters RBC (x1012/l) PCV % HB (g/dl) Hct l/l MCV (fl) MCH (pg) MCHC (g/dl) WBC (x 109 /l) Heterophils % Lymphocytes % Monocytes% Eosinophils % Basophils % JENNINGS 1996 n=70 2.4 31.82 46.30-11.41-53.69 60. monocytes and eosinophils.95-35.13-2.95-3.32 0.13±2.00±11.50 11. 38 0.30 0 0.40 30.50 ---124-147 41.12 0.31 41.63 5.73 ----10.45±0.80-11.40 13.10 WERNERY et al 2004 n=146 3.40-34.28±0.80±0.84 8.16 31.80 58.4 RBC (x1012/l) PCV % HB (g/dl) Hct l/l MCV (fl) MCH (pg) MCHC (g/dl) WBC (x 109 /l) Heterophils % Lymophocytes % Monocytes % Eosinophils % Basophils % Only little information is available on the gyr falcons (Falco rusticolus).21±3.29 ----15.45±0.00-58.59 183.A similar picture shows the literature comparison of the haematology values of saker falcons (Falco cherrug).23±0.93±0.00±1.50 26. heterophils. Table 4: Literature comparison of haematological parameters in saker falcons (falco cherrug) Parameters JENNINGS 1996 n=50 2.64±0.46 4.42 33.10±0. Table 5: Literature data of haematological parameters in gyr falcons (Falco rusticolus) Parameters RBC (x1012/l) HB (g/dl) Hct l/l MCV (fl) MCH (pg) WBC (x 109 /l) Heterophils % Lymphocytes % Monocytes % Eosinophils % Basophils % WERNERY et al 2004 n=187 3.33 0. Major differences can be found in MCV.50 0.40-45.53 61.89 47.90 2.84 60.40±2.72±2.50 0-2 0-0.18±0.74±1.02 1.98 3.16±3.00-42.2 0 81 . D‘ALOIA 1996 n=25 2.50-16.72±3.90 37.46 0. lymphocytes and monocytes.47±0. MCH.96 38-49 11.44 45.17±1.90 3.04 141.70±0.32±5.00±1.50 8.28 15.11±11.45 SAMOUR.78±11.53±12. WBC.78±1.30±0.71±3.50±5.04 139.08 ---15.54-3.54±12. 50 44.27 49.00 37. show already clinical signs of reduced well-being like reduced appetite. 82 . When using the Cell Dyn 3500 analyzer (Abbott Laboratory. (2004) evaluated the gyr hybrid falcons by combining gyr-saker and gyr-peregrine hybrid falcons.43±11.63 1.57±1.40±1. The high values might be related to the use of the Cell Dyn 3500 analyzer (Abbott Laboratory. being the maximum range of WERNERY et al.35 are very rarely evaluated among our patient clientele. 2004).35±0. (2004). As per our observation at the Abu Dhabi Falco Hospital. 10 samples per day.57 52.33±0.38 75.70±3.37±1.12 15.62 46. Due to these unreliable results.23 0 0 Study results n=320 2.55 and 70% heterophils. Table 6: Comparison between haematologicalvalues in the literature and our laboratory results of gyr-Peregrine hybrid falcons (Falco rusticolus x Falco peregrinus) WERNERY et al Parameters RBC (x1012/l) HB (g/dl) Hct % MCV (fl) MCH (pg) MCHC (g/dl) WBC (x 109 /l) Heterophils % Lymphocytes % Monocytes % Eosinophils % Basophils % n=267 3.18±3.75 219.55±2. heterophils and eosinophils (table 6).39±0.36 4.00±4.43 ---9.014 WERNERY et al. not flying well and not gaining weight.24 60.00±2.91±3.26 17.59±8.00±10.14±0. This technical problem resulted in parameters showing very high values that were contradicting clinical conditions of the falcons. RBCs in the range of 3. MCV. the Cell Dyn 3500 analyzer is not able to run the falcon blood with its very thick consistency thus leading to unreliable results due to blockage of the washing function of the analyzer. the Abu Dhabi Falcon Hospital returned to the manual haematology examination.15 45. (2004). WBC and packed cell volume (WERNERY et al. falcons with a WBC of 12.48 7.The value ranges of the gyr peregrine hybrid falcons (Falco rusticolus x Falco peregrinus) differ especially of RBC.18 2.31±3. The results of our study show that the haematological values of both hybrid species are relatively close to each other.45 34. For the examination of more than approx.08 0. yet far below the results measured in WERNERY et al.7±25. North Chicago IL) used for the calculation of RBC. North Chicago IL) we experienced that this equipment can process only a low number of daily falcon blood samples with accurate results.43±1.31±4. WBC. From our patients records hybrid falcons with such elevated blood parameters often display infections or aspergillosis.0 137.9±1. 23 4.11±2.04 Study results GP n=320 2.00 140. 83 . heterophils.75 219.55±2.27 49.63 1. 6: 50 .Table 7: Comparison between haematological values in the literature and our study in gyr-saker hybrid falcons (Falco rusticolus x Falco cherrug) and gyrperegrine hybrid falcons (Falco rusticolus x Falco peregrinus) Gyr hybrid (GS and GP) Parameters RBC (x10 /l) 12 WERNERY et al 2004 GS and GP n=990 3.66 221.14±0.9±1.85±21. D’ALOIA M-A and HOWLETT JC: Normal haematology of captive saker falcons (Falco cherrug).45 34. lymphocytes and eosinophils. (2004) can be observed in hybrid falcons.00±4.83 34.52.33±0. Falco Newsletter No 4. MCV.42±14.57±4. Major differences between our study and WERNERY et al. 2. 2004) which does not produce accurate results as soon as it is used for processing a large number of falcon blood samples per day on one hand. Haematological parameters in captive peregrine falcons (Falco Peregrinus).40±1.36 4.38 75.57 52.92 0.22 49. WBC.59±8. Haematology results as one important piece of the disease picture should never be interpreted alone but always in connection with the clinical disease symptoms of the examined falcons. National Avian Research center.48 7.90±3.37±14. The Middle East Falcon Research Group.13±8. 5.014 GS n=369 2.91 4.26 17.39±0. 1995.15 PCV % HB (g/dl) Hct % MCV (fl) MCH (pg) MCHC (g/dl) WBC (x 10 /l) 9 Heterophils % Lymphocytes % Monocytes % Eosinophils % Basophils % The evaluation of the haematological parameters shows a big variety of value ranges especially of RBC.31 -----14.00±4.7±25.42±1.43±5.50±2.71 ----9.15 60.21±2.91±3.15 --17. Com Haem Inter 1996. These findings may caused by the hand-count method on the other hand and the use of the Cell Dyn 3500 analyzer (Abbott Laboratory.33 44.57 7.68 34. CITATION INDEX 1.40±0.31 0. United Arab Emirates. SAMOUR JH. North Chicago IL) (WERNERY et al.29±0.43±1.42±1.37±1.67±1.10 46.73±1.08 0.24±6. The relatively high ranges in the haematological parameters produced by the Cell Dyn 3500 analyzer were contradicting the clinically healthy picture of the falcons tested.67 0.64 76. DÖTLINGER HS and BIRD DM.50 44.14±0.18±3.58 1.73±3.50 51.88 44. .M. In: FUDGE A. G.O. et al. WERNERY U. (ed): Manual of raptors. Laboratory medicine: avian and exotic pets.Box 45553. 4. 7. In: CAMPBELL T. pigeons and waterfowl. BAILEY T. HOWLETT J. Hannover: Schluetersche Verlagsgesellschaft.H.. United Arab Emirates Email: mmuller@erwda. Muller Dr. Abu Dhabi. 68 . and HARCOURT-BROWN N. KINNE J and SAMOUR J: Colour atlas of falcon medicine. 3-17.gov. Ames. Saunders. MRCVS Abu Dhabi Falcon Hospital/Environmental Research and Wildlife Development Agency (ERWDA) P. 2004: 18. FORBES N. Avian complete blood count. 2000: 9-18. vet.H..A.78.ae 84 . WERNERY R. med. SAMOUR J..A. Iowa: Iowa State Press 1988.W. 5.H. Avian hematology and cytology. Avian hematology.B.. FUDGE A. Haematology.C. Handbook of bustard haematology.M. In: BEYNON P.3. AUTHORS ADDRESS M. 6. Philadelphia: W. CAMPBELL T. National Avian Research Center 1996.B.W. JENNINGS I. Gloustershire: BSAVA 1996.. Infectious diseases . Knowledge of the causes of morbidity and mortality of falcons in the Middle East is essential 85 . Although falcon hospitals have existed in the Middle East for over 25 years. REMPLE 1988. Naldo DVM KEYWORDS Morbidity – Mortality . Fahad bin Sultan Falcon Center. In the past. Kingdom of Saudi Arabia from 1st September 1998 to 31st December 2004 were reviewed to determine the causes of morbidity and mortality amongst falcons used in the sport of falconry in Saudi Arabia. MVZ. This report intends to contribute to the scanty information on morbidity and mortality in captive falcons in the Kingdom of Saudi Arabia and the Middle East as a whole. SAMOUR 1996). Bedouins would trap. J. Riyadh.Viral ABSTRACT The medical records from 12523 falcons from different species presented for examination to the Falcon Specialist Hospital and Research Institute of the Fahad bin Sultan Falcon Center. Dip ECAMS. and hunt with passage falcons during the winter months in order to supplement their basic diet. Kingdom of Saudi Arabia CAUSES OF MORBIDITY AND MORTALITY IN CAPTIVE FALCONS IN SAUDI ARABIA J. they would release the falcons.Falcon Specialist Hospital and Research Center. 1 INTRODUCTION Falcons and falconry have formed an integral part of Arab desert life for thousands of years. Samour. In early spring. L. Arab falconers keep their falcons in air-conditioned rooms or in free-flying aviaries during the moulting months so that they can be used again for the next season. train. The need to maintain falcons in captivity prompted the establishment of falcon hospitals in the different countries of the Arabian Peninsula to cater for their medical needs.Lead toxicosis – Parasitic . REMPLE and GROSS 1993. Nowadays. very little information is available about falcon morbidity and mortality (AL TIMMIMI 1987. RIDDLE and HOOLIHAN 1993.Saudi Arabia – Newcastle . Riyadh. PhD. as caring for birds throughout the summer could strain their already limited resources. Most falcons presented to the Hospital belonged to Saudi falconers. It is our desire that the information presented in this report could contribute to provide the basis for future veterinary research. whole body survey radiographs were obtained in both ventrodorsal and laterolateral positions.6%) hybrid falcons. 394 (3. Selective tissues were collected and preserved in 10% buffered formalin for routine histopathology examination. The falcon examined post-mortem included 329 (69. Kingdom of Saudi Arabia from 1st September 1998 to 31st December 2004 were reviewed. improve therapeutic regimes and to establish suitable preventative medicine programmes. The post-mortem records from 471 falcons that died in the hospital during this period were also reviewed. The main objective of this report is to provide an account of the causes of morbidity and mortality in falcons in the Kingdom of Saudi Arabia (KSA). bacterial and fungal culture and sensitivity testing and cytology.6%) saker falcons (Falco cherrug). Dubai. 37 (7. 31 (6.1%) Barbary falcons (Falco pelegrinoides). All the above analyses were carried out in our own laboratory. Bahrain and the United Arab Emirates (UAE). haematology analyses.8%) gyr. Selective diagnostic procedures included faecal testing for endoparasites.3%) peregrine falcons (Falco peregrinus). 57 (12.6%) lanner falcons.1%) gyr falcons (Falco rusticolus). 1547 (12. In general.1%) peregrine. were also collected and stored at -20ºC for virus isolation. spleen and kidneys. All falcons presented were subjected to a complete physical examination. Riyadh.to understand diseases. The falcon species included 9722 (77. liver. 2 MATERIALS AND METHODS The medical records from 12523 falcons from different species presented for examination to the Falcon Specialist Hospital and Research Institute of the Fahad bin Sultan Falcon Center (FSFC).6%) hybrids and 17 (3. plasma chemistry and blood lead level analyses. Qatar. Falcons examined by endoscopy and radiography were anesthetised with isoflurane administered via a face mask. A comprehensive post-mortem examination was carried out in all deceased falcons within 24 hours of death.8%) saker. 576 (4.1%) lanner falcons (Falco biarmicus) and 17 (0. 86 . Swabs from affected organs were collected for microbiology examination and examined in our own laboratory. including brain. endoscopy and radiography examinations. In selected cases tissue samples. but an unknown number of falcons originated from Kuwait. UAE. Histopathology and virus isolation analyses were carried out at the Central Veterinary Research Laboratory. 267 (2. toxicosis and metabolic or nutritional diseases (Table 3). The classification of the causes of morbidity was based on the primary medical condition identified on clinical examination (Table 3). The number of falcons deceased at the Hospital is shown in Table 4. The most common cause of mortality was Newcastle disease followed by septicaemia. The incidence of some diseases varied during the study period due to improved health awareness and improved diagnostic techniques. followed by traumatic injuries. visceral gout. In contrast only four cases of lead toxicosis were recorded in 2004. In addition. to 44 in 2003 and 42 in 2004. In addition. This number increased to 14 in 2002.3 RESULTS The total number of falcons presented for examination is shown in Table 1 and a breakdown of the total number of falcons per species is shown in Table 2. severe trauma and sour crop (Table 5). For instance. This was the result of increased awareness of the two clinical presentations of Newcastle diseases in falcons. The causes of mortality were based on the primary pathologic lesion found at post-mortem examination (Table 5). every effort should be made to confirm the diagnosis by using either in-house laboratory facilities or commercial laboratories. 87 . the most common causes of morbidity were infectious diseases.5 years (NALDO and SAMOUR 2004). notably Newcastle disease. In a preliminary study for example. the neurotropic and viscerotropic forms (SAMOUR and NALDO 2004) and routine virus isolation analysis from every suspected case of ND. Knowledge of the most common diseases affecting falcons in the region could provide a better understanding of the diseases and effective ways to prevent them. the number of lead toxicosis cases diagnosed at the Hospital decreased significantly over the six year period due to public awareness campaigns and a better understanding of the toxic effects of lead. only one case of Newcastle disease (ND) was diagnosed at post-mortem in 2001. The need to keep and maintain a medical record database cannot be over-emphasised. Nevertheless. 96 cases of lead toxicosis were recorded over a period of 2. aspergillosis. It is hoped that the information presented in this paper could serve as a base for other similar studies in the region. 4 DISCUSSION It is beyond the scope of this paper to discuss in detail the different causes of morbidity and mortality in falcons during the study period. amyloidosis/hepatitis. 05%) 2001 1886 (81%) 291 (12.6%) 2 (0.3%) 270 (11.7%) 60 (2.5%) 17 (1.9%) 2 (0. of visits 420 1352 2880 4321 4113 4483 5957 23526 Table 2.6%) 45 (1.8%) 119 (5.Table 1.9%) 0 2340 2002 1599 (76%) 242 (11.6%) 1 (0.5%) 79 (3.5%) 60 (2.4%) 58 (2. Number of falcons per species presented for examination at the Fahad bin Sultan Falcon Center from 1st September 1998 to 31st December 2004 Species Saker Peregrine Gyr Hybrid Lanner Barbary a 1998 282 (89%) a 27 (8.6%) 1 (0.4%) 163 (6.6%) 0 1999 740 (81%) 132 (14.1%) 2101 2003 1743 (74.2%) 2676 314 912 1834 TOTAL Percentage of the total number of birds.5%) 104 (4.5%) 8 (0.0%) 6 (0. of new falcons 314 863 1684 2046 1675 1655 1656 9893 Total no. 88 .9%) 58 (2.2%) 187 (7%) 55 (2.7%) 30 (1.9%) 0 2000 1505 (82%) 237 (13%) 29 (1. Total number of falcons presented for examination at the Fahad bin Sultan Falcon Center from 1st September 1998 to 31st December 2004 Period 1998 1999 2000 2001 2002 2003 2004 TOTAL Total no.3%) 2 (0.3%) 2346 2004 1967 (73.6%) 32 (1. of falcons 314 912 1834 2340 2101 2346 2676 12523 No.5%) 348 (13%) 113 (4.5%) 9 (1%) 14 (1. Coli. fractures g Ring constriction.Undetermined hepatophaty Traumatic injuries Bumblefoot Collision-type injuriesf Gunshot injuries Eye and eyelid abnormality Bursitis Miscellaneous traumag Toxicosis Lead Ammonium chloride Pharmacological toxicosis Metabolic or nutritional diseases Low condition Gout Sour crop GIT obstruction or impaction Uropygial gland abnormality Moulting abnormality Metabolic bone disease Hernia Pancreatic abnormality Degenerative disease Amyloidosis Neoplastic disease Lipoma a b No of cases 5. dislocation. abrasion 89 . distal digital necrosis. Shigella sp.151 285 380 398 901 265 355 183 15 293 7 234 207 49 15 68 25 31 59 17 102 10 5 7 15 3 Endoparasites. air sacculitis. trichomoniasis. burns. Causes of morbidity diagnosed in falcons at the Fahad bin Sultan Falcon Center from 1st September 1998 to 31st December 2004 Categories Infectious diseases Parasitica Bacterialb Viralc Fungald Undetermined respiratory diseasee …. Salmonella sp. avian pox.. herpesvirus hepatitis. Klebsiella sp. aeruginosa. pneumonia f Avulsion. candidiasis e Rhinitis. haematozoa E. Proteus sp. rhinoliths. P. c Newcastle disease. detached claws.. C..Table 3. avian influenza d Aspergillosis. perfringes. lacerations. 7 - 2003 No 44 12 4 10 10 10 3 4 3 16 7 123 % 35.6 34.1 8.1 2.Table 4.8 4.7 7.4 8.8 6.4 - No 1 5 4 2 14 1 2 13 42 % 2.5 4. candidiasis. 90 .1 2.4 5. c Miscellaneous diseases included lower GIT impaction.0 15.6 6.0 12.1 8.8 4. Causes of mortality diagnosed in falcons at the Fahad bin Sultan Falcon Center from 1st January 1999 to 31st December 2004 1999 Causes Newcastle disease Aspergillosis Severe trauma Amyloidosis /hepatitis Visceral gout Septicaemiaa Bumblefoot (severe) Lead toxicosis Sour crop Other infectious diseasesb Miscellaneous diseasesc Total a b 2000 No 3 6 6 1 8 24 3 4 3 6 5 69 % 4.8 3.1 9.7 - 2004 No 42 9 4 9 6 7 3 1 2 3 5 91 % 47.8 31 - Causes of septicaemia included Escherichia coli.7 11.3 4.6 2.8 25. Table 5.6 - 2004 53 10 11 14 3 91 % 58.9 5. inhalation pneumonia.3 - Primarily gyr-sakers and gyr-peregrines.4 13.4 3.0 4. avian influenza.9 4.3 1.2 11.3 18.4 4.5 11.7 7.9 9.1 1.9 7.7 9.4 8.2 - 2001 No 1 8 2 2 3 11 2 1 6 8 44 % 2. Number of falcons deceased per species at the Fahad bin Sultan Falcon Center from 1st January 1999 to 31st December 2004 Species Saker Peregrine Gyr Hybrida Lanner Total a 1999 36 3 1 2 42 % 85.3 8.4 15.7 1.2 3.4 8.3 2.9 2.7 12.6 18.0 5. falcon pox.3 4.7 3.4 9.5 - 2002 69 19 6 4 4 102 % 67.4 11.8 9. pharmacological toxicosis.3 4.2 9.6 4.9 4. scorpion sting. Other infectious diseases included herpesvirus hepatitis. trichomoniasis.7 4.9 3.9 - 2003 85 12 14 10 2 123 % 69.9 8.3 13.8 - 2001 34 2 3 3 2 44 % 72. tuberculosis.8 - 2000 52 11 2 4 69 % 75.2 - 2002 No 14 10 7 8 9 13 5 5 5 13 13 102 % 13.9 3.8 6. ammonium chloride toxicosis.6 7.9 4.9 12. low body condition.2 4.9 6.6 18.8 33. pseudomoniasis.7 8.5 6.6 5.4 3.8 4.1 2.3 2. Clostridium perfringens and Salmonella species. pancreatic atrophy.8 12. 830.O.239. SAMOUR JH and NALDO JL.193. 2. Causes of morbidity and mortality in falcons in Saudi Arabia. (eds): Peregrine Falcon Populations: Their Management and Recovery. RIDDLE KE and HOOLIHAN J. 7. Proc Assoc Avian Vet. PhD. Dip ECAMS Fahad bin Sultan Falcon Center P. 4. In: CADE TJ. operation and admissions summary. 131 . Mohammed Shiyan. Minneapolis: University of Minnesota Press. J Avian Med Surg 2004. Orlando 1996: 233 . MVZ. COOPER JE.133. NALDO JL and SAMOUR JH. Falcons and Falconry in Qatar. Falconry and Birds of Prey in the Gulf. Wilson Joseph. AL TIMIMI FA. 5. Proc Ass Avian Vet. 55. 1993. 1987. United Arab Emirates: Motivate Publishing. 825 .5 ACKNOWLEDGEMENTS The authors would like to thank HRH Prince Fahad bin Sultan bin Abdulaziz Al Saud for continuing support to the clinical and research programme of the Hospital and to Dr Niel Nuguit. THELANDER CG and WHITE CM. 1988.241. An overview of Arab falconry. Newcastle disease in falcons in Saudi Arabia. New Orleans 2004. falcons and falconry in the Middle East. REMPLE JD and HUNTER DB (eds): Raptor Biomedicine. Doha: Ali bin Ali Printing Press. Veterinary medicine. 18(4): 229 . SAMOUR JH. Dubai. A research hospital for falcons: design. Mohammed Foaud. 1993. ENDERSON JH. AUTHOR ADDRESS Jaime Samour. In: REDIG PT. REMPLE JD and GROSS C. Boise: The Peregrine Fund Inc.com 91 . Salem Al Yami and Merle Apo for help with the practice statistics. 6. 188 . REMPLE JD. its medical lore and the introduction of avian medicine in the Arabian Gulf. 3. Riyadh 11322 Kingdom of Saudi Arabia Email: falcon@shabakah. 6 CITATION INDEX 1. United Arab Emirates HISTOPLASMA SP.F10 ABSTRACT Histoplasmosis is an infectious. but not contagious mycotic disease of animals that has been reported in a wide variety of wild and captive mammals and occasionally in nondomestic birds. treatment and outcome of nine falcons with lower respiratory tract disease from which H. Histoplasma sp.Histoplasma sp Voriconazole . VAUGHN 1996. Histoplasma capsulatum. diagnosis.1%) from 116 fungus positive airsac biopsies that were collected over a one year period at the Abu Dhabi Falcon Hospital. mycology records from two falcon hospitals were analysed. C. Our observations demonstrate that Histoplasma sp.Dubai Falcon Hospital1. E. Histoplasma sp. .Histoplasmosis . Al Ain.Fungal infection . was isolated once (prevalence of 1. pigeons and poultry (BAUCK 1994. was isolated 6 times (prevalence of 5.INCIDENTAL FINDING. Silvanose1. The causative agent of histoplasmosis is a dimorphic fungi. humid areas of the world (BUREK 2001). OR OCCASIONAL OPPORTUNISTIC PATHOGEN OF FALCONS IN THE MIDDLE EAST ? T. Di Somma1 KEYWORDS Falcon .Aspergillosis . The 92 . We reviewed the clinical findings. Dubai. 1 INTRODUCTION Histoplasmosis is an infectious. Project for Falcon Conservation2 (Profalcon). A. in isolates from airsac biopsies. but not contagious mycotic disease that has been occasionally reported in non-domestic birds. Bailey1. which occurs naturally in soil in warm.2%) from 86 fungus positive airsac biopsies that were collected over a one year period at the Dubai Falcon Hospital. BUREK 2001). although its significance remains unclear. Pesci2. capsulatum was isolated alone or as part of a mixed infection with Aspergillus sp. In order to determine the prevalence of Histoplasma sp. is a rare isolate from falcons with lower respiratory tract disease. UNDERDIAGNOSED. pigeons and poultry. benign. was isolated from the lower respiratory tract of eight falcons seen by the authors at the Abu Dhabi Falcon Hospital [ADFH] (2000-2001) and the Dubai Falcon Hospital [DFH] (2002-2003). The fungi were identified by culture appearance and morphological characteristic under microscope with the help of lactophenol aniline stain (QUINN et al.2 Bacteriology and mycology Biopsy samples were cultured in sheep blood agar. 2) faecal and crop parasitology. 2. loss of weight (2). Two birds were admitted for pre-purchase examinations and no clinical history was available. diagnosis. Impression smears on slides were air dried. 2 MATERIALS AND METHODS 2. Infection of the respiratory tract occurs when dust containing spores are inhaled. The samples were obtained directly with biopsy for culture and cytology. In order to determine the prevalence of Histoplasma sp. poor exercise performance (3) and dyspnoea (1). One bird was admitted for a premoult health check and no clinical signs had been noted by the owner. in isolates from airsac biopsies. fixed and stained with Neat stain (Astal Diagnostics. capsulatum was isolated alone or as part of a mixed infection with Aspergillus sp. progressive and potentially fatal disease. 93 . Clinically apparent histoplasmosis can be a localised. was included in the study. The birds were females. Our retrospective study reviews the clinical findings.organism lives as a soil saprophyte. particularly where animal faeces accumulate (WALSH et al. 2002). followed by lymphohaematogenous dissemination via the monocyte-macrophage system. or a disseminated. mycology records over a one year period from January 2000 to January 2001 at the ADFH and from June 2003 to June 2004 from the DFH were analysed. The birds were given a clinical examination that included: 1) physical examination. 1995). During endoscopy biopsy samples were collected from affected areas of the respiratory system of each individual.1 Clinical history and investigations Histoplasma sp. vomiting (1). In mammals the disease is characterised by primary and secondary foci of infection becoming established in the lungs. inappetance (1). 7 birds were juveniles and 2 were adults. 5) survey whole body radiographs and 6) endoscopic examination of the lower respiratory tract. Blood agar plates were incubated at 37°C in a 5% CO2 incubator for 48 hrs and MCA plates were incubated aerobically at 37°C for 48 hrs. 3) haematology. The SA plates were incubated at 37°C for 48 hrs and further 8 to 14 days at room temperature (25 °C). USA). A further bird with airsacculitis and cytological findings consistent with Histoplasma sp. treatment and outcome of nine falcons with lower respiratory tract disease from which H. pulmonary disease. 4) plasma biochemistry. Six affected falcons were presented with a history of. MacConkey’s agar (MCA). for bacteriology and Sabouraud’s agar (SA) for mycology. radiological findings included air sac consolidation and splenomegaly. only. and from which Aspergillus sp. only. The radiological abnormalities included prominent bronchial pattern. 2) weight gain. serratospiculiasis (1) and trematodes (1). clinical outcome and follow-up The birds were seen over a four year period at two hospitals by 3 veterinarians. inflammatory cells including heterophils.3 Treatments. opaque and vascularised air sacs (6). 3 RESULTS 3. and from which Aspergillus sp. Cytological findings included. In the 3 cases that were cytology and/or culture positive for Histoplasma sp. Haematology parameters were within normal ranges in one bird. bulb like structures) and /or septate hyaline hyphae consistent with Aspergillus sp. Radiographic abnormalities were present in the two of the three DFH birds. hepatic inflammation (1).7 x109/l) with mild to marked heterophilia.1 Clinical investigations Physical examination findings included poor pectoral muscle condition (5). was not isolated. follow-up was possible at 3 and 12 months in one bird and at 5 months in another. endoscopy findings included opaque and vascularised air sacs (2). Follow-up procedures included the examination previously described. thickened. small (<5mm) multifocal white to yellow granulomas (3).7-22. visible caudal airsac line. bumblefoot (1). large disseminated granulomas (>5mm) (1). trematodes and Serratospiculum sp. The different treatment regimens are summarised in Table 1. macrophages.2. In all 9 cases endoscopy revealed a generalized air sacculitis including. 4) negative culture or cytology of endoscopic biopsies. infection (4). After discharge. Treatment ended and birds were discharged from the hospital according to the following criteria: 1) resolution of the presenting signs. 3) marked improvement of the lower respiratory tract lesions during endoscopy. diffuse cheesy plaques (6). No abnormalities were noted in 4 birds. an inflammatory response in the air sacs lining cells including vacuolation.) were treated. Parasitic infections (coccidia. extensive. focal densities within the lungs or the air sacs and splenomegaly. and 5) haematology within normal ranges. low pectoral muscle tone (4) and dehydration (2) in 5 birds.2 Laboratory investigations Haematology results from 8 of 9 birds showed from mild to severe leukocytosis (6. hepatomegaly (1) and pulmonary congestion (1). 3. the presence of fungal spores (coniodospores. giant cell formation and metaplasia (4) and the 94 . asymmetry of the air sacs because of air sac consolidation or hyperinflation. multinucleated giant cells (6). Other underlying diseases diagnosed included: coccidiosis (2). diffuse cheesy plaques (1) and small (<5mm) multifocal granulomas (1). was not isolated. In one of these 3 cases that was cytology positive for Histoplasma sp. was isolated from airsac biopsy specimens from 8 of 9 cases. fumigatus (40). niger (13). was isolated 6 times (prevalence of 5. 2) the birds had gained weight (7-13% from time of presentation). A. fumigatus (1). Candida albicans (1) and Mucor sp. One of these birds was examined 12 months later. After 10 days of incubation. A. 5) cytology biopsies were negative and 6) haematology results were within normal ranges. niger (9). terreus (12). A. A. A. Histoplasma sp. fumigatus (45). Scediospium sp.1%) from 116 fungus positive airsac biopsies that were collected from falcons over a one year period from 1/1/00 to 1/1/01 at ADFH. nidulans (2). Other fungi isolated at ADFH comprised. A. flavus (45). nidulans (1) and A. including endoscopy and were considered fully clinically recovered. 3. averaging 1-2µm in diameter with small lollipop-like microconidia. up to 4µm in size. Both birds had a complete examination. was the only organism isolated in 2 birds. niger (1). but bacterial and fungal cultures were negative. A. Five hospitalised cases were discharged after 21-70 days (median 36 days) treatment. Two of the four birds discharged after what was considered to be a full course of treatment were both presented for follow-up examination at 3 and 5 months after first diagnosis. No bacteria were isolated from biopsy specimens. were isolated. Microscopically the hyphae are delicate. 95 . 3) endoscopic examinations revealed marked improvement of the surface of air sacs and lung. consisting of a central spherical basophilic body surrounded by an unstained halo. Histoplasma sp. and Aspergillus sp. (7). A further case was improving clinically. Other fungi isolated at DFH comprised. In one bird the strain of Aspergillus was not determined.3 Treatments and follow-up A summary of the treatment regimens. fungal colonies appeared white to cream in colour with cottony ariel hyphae. In these five birds: 1) the presenting clinical signs had resolved. clinical response and follow-up are presented in Table 1. flavus (1) and A. Histoplasma sp. In 6 birds mixed cultures of Histoplasma sp. A. Histoplasma sp. (1). (2). was isolated once (prevalence of 1. Peciliomyces sp. but was removed from the hospital against medical advice after 10 days treatment by an owner who wished to train his bird and go hunting. In one bird organisms consistent with Histoplasma were seen in cytology preparations. A. versicolor (1). Two cases from which Histoplasma sp.2%) from 86 fungus positive airsac biopsies that were collected from falcons over a one year period from 1/6/03 to 1/6/04 at DFH. nidulans (1). A. The strains of Aspergillus included A. 4) no bacteria and fungi were cultured from biopsies. A.presence of multinucleated giant cells containing globular structures. flavus (14). no abnormalities were detected and the falconer reported that the bird was flying well. were isolated died 7 and 36 days after hospitalisation. morphologically consistent with Histoplasma organisms (3). Unfortunately the owners would not allow post-mortem. Aspergillus sp. Another case treated as an outpatient was not seen again. and Aspergillus sp. (3). 4 DISCUSSION In the majority of cases (66%) described in this report Histoplasma sp. and Aspergillus sp. were isolated together and it is not possible to relate pathology to either organism. Only in two cases was Histoplasma sp. isolated as the sole organism from airsac biopsies and both birds showed signs of airsacculitis. However, in these cases the failure to culture Aspergillus sp. does not conclusively rule out concurrent infection with aspergillosis. It is not uncommon to fail to culture Aspergillus sp. from biopsies collected from cases with endoscopically apparent lower respiratory tract disease, and where cytology of biopsies is consistent with a fungal infection. It is known that some strains of Aspergillus (e.g. A. fumigatus) produce toxins (e.g. gliotoxins) that are highly immunosuppressive (WERNERY et al. 2004). It is possible that a primary aspergillosis infection could predispose falcons to infection with other pathogens, such as Histoplasma sp. Another interesting finding is the wide range of fungi that are isolated from falcons with lower respiratory tract disease (Table 2). Some isolates (e.g. A. terreus) are recognized pathogens in humans and animals. Other isolates (e.g. Scediospium sp.) are considered emerging pathogens in human medicine or as atypical causes of infection in humans and domestic animals (e.g. A. nidulan, Mucor sp.). Studies to assess if these less commonly isolated fungi are clinically relevant or are simply contaminants are warranted. In humans it is estimated that more than 95% of cases of histoplasmosis are self-limiting and the small number of infection episodes that advance to overt disseminated disease are associated with predisposing risk factors, particularly defects in cell-mediated immunity (WALSH et al. 1995). In wild animals, additional immunosuppressive factors including the stress of capture, transportation, confinement and altered habitat are thought to trigger overt histoplasmosis (BUREK 2001). Fungal infections in falcons are similarly thought to occur secondary to an immunosuppressive event. In the Middle East, aspergillosis typically occurs in captive bred birds shortly after they arrive from Europe and North America and are exposed to high temperatures, high humidity, new environments and falconry training methods. The various stresses associated with these changes contribute to predisposing birds to systemic fungal infections. The Histoplasma organism thrives in soil mixed with faeces and especially hen houses and bird/bat roost. In humans, outbreaks of histoplasmosis are seen in people who have demolished buildings infested with bird guano. The frequent finding of Histoplasma organisms in association with bird droppings is considered to be due to environmental enrichment by the bird faeces rather than propagation and transmission by birds (BUREK 2001). Decreasing exposure to substrates enriched with bat or bird droppings is important in minimising infection with Histoplasma sp. Spraying infected soil and faecal deposits with 3% formalin destroys some spores. Histoplasmosis has zoonotic potential and in humans may cause pneumonitis that progress to a disseminated disease of the reticuloendothelial system (BAUCK 1994). Clients should be warned of the risks of the disease, because the environment where the bird was exposed may be a risk to humans. The cases reported in this study could have been linked to poor hygienic conditions in the environment where the birds were housed. 96 Cryptococcus has been more widely reported as a cause of disease in psittacines and pigeons and BAUCK (1994) reports that histoplasmosis is similar to cryptococcosis in many ways, but has been less commonly reported in birds. Some authors consider that histoplasmosis is not a clinical problem in birds because the body temperature of birds is too high to support the growth of this mycotic organism (CARPENTER and GENTZ 1997). While our observations are far from comprehensive, with no post-mortem/histopathology examinations conducted on the dead birds, they do demonstrate that Histoplasma sp is a rare isolate from falcons with lower respiratory tract disease. It is possible that this condition is underdiagnosed because in routine SA cultures it is easily overgrown by other fungi such as Aspergillus sp. and because it is slow growing, requiring 2-4 weeks of incubation (QUINN et al. 2002). Aspergillus sp. cultures usually grow in less than 5 days at 37C and it is routine to discard SA plates after 4-5 days incubation. The use of deep-filled culture plates or enriched media (brain heart infusion) may improve the chances of culturing this organism. Table 2 presents minimum inhibitory concentrations (MIC) taken from the literature of two commonly used antifungal agents against H. capsulatum and for comparative purposes against A. fumigatus (PRESCOTT 2000). The MIC of antifungal agents against H. capsulatum are 4-5 times lower than the equivalent MICs for A. fumigatus, suggesting an increased susceptibility of Histoplasma to these drugs. In our study a positive clinical response was seen in cases given itraconazole, itraconazole and F10, itraconazole and amphotericin B and voriconazole and amphotericin B. Itraconazole has been successfully used to treat cats with histoplasmosis (HODGES et al. 1994). Recurrences occurred in the cats after completion of therapy requiring a further 2-3 months of treatment. Antifungal regimens in humans are often extended for 6-12 months and in veterinary species treatment duration recommendations vary from 2 to 6 months and one month past the resolution of the disease. Unfortunately, client compliance in outpatients with extended antifungal therapy is difficult in the Middle East and extended therapies, while desirable, may not always be realistically achievable. At the current time we must leave unanswered the title of our talk, the question of whether Histoplasma sp. is an incidental finding, underdiagnosed, or just an occasional opportunistic pathogen of falcons in the Middle East? Further investigations including histopathological and immmunohistopathology diagnosis from confirmed cases are needed to conclusively demonstrate the clinical significance of this interesting fungus that has been isolated from falcons. 5 ACKNOWLEDGEMENTS We thank H.H. Sh. Hamdan bin Rashid al Maktoum and Mr Humaid Obaid al Muhairi, Dubai Falcon Hospital Director, for their support of the work of Dubai Falcon Hospital and all of the falcon hospital team for their assistance with cases. We thank H.E. M. Al Bowardi for his support of the veterinary science programme of Abu Dhabi Falcon Hospital. 97 6 CITATION INDEX 1. 2. 3. 4. 5. 6. 7. 8. 9. BAUCK L. Mycoses. In: RITCHIE BW, HARRISON GJ, HARRISON LR. (eds): Avian Medicine: Principles and Application. Lake Worth, Florida: Wingers Publishing Inc 1994; 997 - 1006. BUREK K. Mycotic disease. In: WILLIAMS ES, BARKER IK. (eds): Infectious Diseases of Wild Mammals. London: Iowa State University Press 2001; 518 - 520. CARPENTER JW and GENTZ EJ. Zoonotic diseases of avian origin. In: ALTMAN RB et al. (eds): Avian Medicine and Surgery. Philadelphia: W.B. Saunders Co, 1997; 350 - 363. HODGES RD, LEGENDRE AM, ADAMS LG, et al. Itraconazole for the treatment of histoplasmosis in cats. J Vet Int Med 1994; 8: 409 - 413. PRESCOTT JF. Antifungal chemotherapy. In: PRESCOTT JF et al. (eds): Antimicrobial Therapy in Veterinary Medicine. Iowa: Iowa State University Press, 2000; 367 - 395. QUINN PJ, CARTER ME, MARKEY BK, CARTER GR. (eds): The dimorphic fungi. In: Clinical Veterinary Microbiology. London: Mosby International Ltd 2002; 402 - 408. VAUGHN S. What is your diagnosis? J Avian Med Surg 1996; 10: 37 - 39. WERNERY R, WERNERY U, KINNE J, SAMOUR J. Colour Atlas of Falcon Medicine. Hannover: Schlutersche, 2004; 134 WALSH TJ, MITCHELL TG, LARONE DH. Histoplasma, blastomyces, coccidioided, and other dimorphic fungi causing systemic mycoses. In: MURRAY PR (ed): Manual of Clinical Microbiology. Washington: ASM Press 1995; 749 - 764. AUTHORS ADDRESS Tom Bailey, BSc, BVSc, MRCVS, Cert Zoo Med, MSc (Wild Animal Health), PhD, Dip ECAMS Dubai Falcon Hospital PO Box 23919 Dubai, United Arab Emirates Email:
[email protected] 98 Central Veterinary Research Laboratory1, Dubai, Dubai Falcon Hospital2, Dubai, United Arab Emirates MYCOBACTERIOSIS IN HUNTING FALCONS IN THE MIDDLE EAST U. Wernery1, DVM, PhD, F.A. Hassan1, MSc, T. Bailey2, BVSc, MSc, PhD, Dip ECAMS, CertZooMed, B. Johnson1, MSc, J. Kinne1, DVM KEYWORDS Falcon – Mycobacteriosis – Mycobacterium – Culture – PCR ABSTRACT One thousand two hundred and three falcons were tested for mycobacteriosis. The samples originated from 702 falcons necropsied at Central Veterinary Research Laboratory (CVRL), from 200 falcons necropsied outside CVRL, and 301 biopsies. In total 23 (1.9%) falcons were diagnosed with avian mycobacteriosis due to typical granulomas and positive Ziehl-Neelsen stains of histological slides. The organs mainly infected were liver followed by spleen and peritoneum. Due to the unavailability of PCR and culture facilities at the beginning of our investigations, only a small number of samples were tested using PCR and culture. PCR was able to detect DNA from nontuberculous mycobacteria from formalin fixed, paraffin embedded tissue samples in 2 out of 10 cases. PCR from strains cultured on Loewenstein/Jensen or Herrold’s egg yolk slant medium identified strains as M. avium paratuberculosis (2x) and M. avium (2x). Eight falcons with mycobacteriosis were examined at Dubai Falcon Hospital. Clinical signs as well as results of radiographs, endoscopy and haematology are presented. 1 INTRODUCTION Mycobacteriosis is a common bacterial disease among wild and particularly captive birds of prey (HEIDENREICH 1995) with a worldwide distribution. The disease has been called avian tuberculosis, but classical tuberculosis lesions are only one of the many manifestations of mycobacterial infection in birds. Therefore, mycobacteriosis is a more appropriate term for this disease (TELL et al. 2001). Several mycobacterial species can cause the disease in birds, principally Mycobacterium (M.) avium, M. intracellulare and M. genavense. As M. avium and M. intracellulare share some common antigens, these species are often grouped and termed as M. avium- 99 intracellulare complex (MAI) (GRANGE et al. 1990). Sometimes M. avium subsp. paratuberculosis and M. lepraemurium which rarely cause avian mycobacteriosis are also included in the MAI complex because of their close relationship to M. avium. Other mycobacterial species, belonging to the M. tuberculosis complex (MTC, LACHNIK et al. 2002), may also cause lesions in birds including M. tuberculosis and M. bovis (VAN DER HYDEN 1997). The disease becomes apparent in a variety of clinical forms, and the bacteria remain infectious in the environment for many months. This paper describes the diagnosis of mycobacteriosis in hunting falcons from the Middle East. 2 MATERIAL AND METHODS A clinical and laboratory study was performed on falcon mycobacteriosis at Dubai Falcon Hospital (DFH) and Central Veterinary Research Laboratory (CVRL). Over a period of 9 years (1995 – 2004) 702 falcons were necropsied and tuberculosis suspicious organ samples stained according to Ziehl-Neelsen (ZN). Furthermore, organ samples from 200 falcons necropsied outside of CVRL in different countries of the Middle East and 301 biopsies collected from sick falcons were also included in this study, totalling 1,203 falcons. Clinical investigations were performed on 8 falcons, suspicious for mycobacteriosis including physical examination, endoscopy and haematology before being euthanased due to poor prognosis. Besides histological investigation, culture was performed on 4 cases using LowensteinJensen (LJ) medium slants (BBL, BD221387) and Herrold’s egg yolk (HEY) slants with mycobactin J (BBL, BD222233) after decontamination and concentration of the samples according to TELL et al (2003). The suspension was centrifuged for 20 minutes at 1500 rpm, the supernatant discarded and the sediment used as inoculum. The LJ and HEY slants were incubated at 37oC for 6 – 8 weeks and inspected weekly for any growth. For DNA extraction 1 suspicious colony was picked from the LJ or HEY agar and suspended in 1 ml of sterile distilled water. This suspension was boiled for 30 min. Extracted DNA from suspected colonies and from fresh tissue was processed using High Pure PCR Template Preparation Kit (ROCH 1796828) according to the manufacturer’s recommendation. Furthermore, DNA was extracted from formalin fixed, paraffin embedded tissue samples (airsac, liver, spleen, intestines) of 10 falcons according to CHRISTOPHE et al. (2000). For PCR amplification three pairs of primers were used according to VITALE et al. (1998). PCR was performed on 10 ZN-positive falcons using formalin samples, as well as on fresh tissue samples from 4 out of these 10 falcons. Furthermore, 4 strains cultured on LJ agar were analysed by PCR. 100 3 RESULTS Twenty-three cases (1.9%) of falcon mycobacteriosis from a total of 1,203 birds were diagnosed by histology from 1995 to 2004 at CVRL. Acid fast rods were found in 8 of 702 necropsies, in 10 of 200 organ samples received from outside, as well as in 5 of 301 biopsies through ZN staining of histology slides. A steady increase of mycobacteriosis in hunting falcons was noticed with a single case in 1995, five cases in 2000, eight cases in 2003, and six cases in 2004. The species mostly involved were saker falcons (9), gyr x saker hybrids (4) followed by gyr falcons (3), gyr hybrids (3), and peregrines (3). Gross pathology of carcasses and organs of all 23 tuberculosis cases revealed yellow-whitish caseous nodules 1 to 4 mm in diameter in the liver (17), spleen (14), peritoneum (7), skin (5), lung (4) and kidney (2). In airsac and heart, tuberculous lesions were only seen once. The granulomas in peritoneum, airsac, heart and skin were diffuse. Acid-fast rods were observed by histopathological examination in all macroscopic suspicious organs. Histopathology revealed central necrotic granulomas with numerous intralesional acid-fast rods surrounded by giant cells. No mineralisation of the lesions was seen. Suspicious skin biopsies showed chronic granulomatous-necrotizing dermatitis containing some acid-fast rods. In one of these cases the lesion was found in the conjunctival-lacrimal gland. Concurrent aspergillosis was diagnosed in 12 of 18 ZN-positive cases from necropsied falcons. In 9 of these 18 cases systemic amyloidosis was also found during histopathological investigation. Eight tuberculous suspicious falcons were examined at DFH. The following clinical signs were observed: Loss of weight (3), inappetance (2), vomiting (1), change in faecal consistency (2), biliverdinuria (2), blood in the faecal (1), depression (1), poor exercise performance (1) and dyspnoea (3). Physical examination findings included poor pectoral muscle condition (4), the presence of palpable subcutaneous nodules in the neck/crop region (1), carpal bursitis (2), an enlarged hardened liver (1), ascites (1) and dehydration (2). Other underlying diseases diagnosed included: Trichomonosis (1), Caryospora sp. (3), Porrocaecum sp. (1), Chlamydophilosis (1), lead toxicosis (1), bumblefoot (1), serratospiculiasis (1), Syngamus sp. (1) and candidiasis (1). Haematology results from 5 birds showed moderate to severe leucocytosis (from 15.5 to 57.5 x 109/l) with marked heterophilia and moderate to severe monocytosis. All birds were anaemic with low packed cell volumes and haemoglobin values. Radiographs from four cases were taken. Radiographic abnormalities were present in all four investigated birds and included: multiple granulomas in the crop and neck (1), hepatomegaly (3), radiodensities within the alimentary tract (1), focal densities within the lungs or the air sacs (2), a lead pellet (1) and splenomegaly (1). Five cases were endoscoped. Endoscopy revealed hepatomegaly (2), pulmonary inflammation (1), generalized airsacculitis including; thickened, opaque and vascularised air sacs (1), small (< 5 mm) multifocal white granulomas in the surface of abdominal organs and liver (3), pus oozing from the ostium (1), fibrinous adhesions (1) and splenomegaly (1). 101 Mycobacteria-suspicious colonies from 4 falcons (Table 1) grew 4 – 6 weeks after inoculation on LJ or HEY slants, which stained acid fast. Primary isolation of M. avium paratuberculosis was possible on HEY slant from one case. However, subsequent subcultures grew on LJ slants as well. The isolates from the other three cases (Table 1) grew on both agars. PCR did not detect mycobacterial DNA from fresh tissue samples in 4 ZN-positive falcons and identified only 2 of 10 formalin fixed ZN-positive specimens belonging to the non-tuberculous complex (NTC). PCR identified 2 isolates from culture as M. avium paratuberculosis and 2 as M. avium (Table 1). Table 1. Results of culture and PCR from 10 ZN-positive falcons. Primary culture Species Peregrine Saker Peregrine Gyr hybrid Saker Gyr Saker Gyr x Saker Saker Peregrine Sample n.r. n.r. n.r. n.r. Spleen Intestine Liver n.r. Liver n.r. Airsac LJmedium Negative Positive Positive Positive HEYmedium Positive Positive Positive Positive Formalin fixed tissue NTC* Negative Negative Negative Negative Negative NTC* Negative Negative Negative PCR Fresh tissue n.r. n.r. n.r. n.r. Negative Negative n.r. Negative n.r. Negative Culture n.r. n.r. n.r. n.r. M. avium paratuberculosis M. avium n.r. M. avium n.r. M. avium paratuberculosis * NTC = non-tuberculous complex nr = not received 4 DISCUSSION From 1,203 falcons tested at CVRL since 1995, 23 (1.9%) cases of mycobacteriosis were diagnosed. The diagnosis was based on typical tuberculous granulomas, and the detection of acid-fast rods in histology sections using ZN staining. Mycobacterial strains grown on LJ and HEY agars were identified by PCR as M. avium (2x) and M. avium paratuberculosis (2x). Also, with PCR, mycobacteria were identified from a peregrine and a saker falcon from formalin fixed and paraffin embedded tissues as belonging to the non-tuberculous complex (NTC) which also includes M. avium, M. 102 intracellulare and M. avium paratuberculosis (LACHNIK et al. 2002, CHRISTOPHE et al. 2002). PCR has clearly identified the cultivated mycobacterial strains. It is not completely clear why PCR failed to identify mycobacterial DNA from 4 fresh and 8 formalin fixed tissues. One explanation could be the sample size. The bigger parts of the small granulomas were used for histology and culture, leaving little material for PCR. At the beginning of our diagnostic procedure the PCR used was also limited to identifying only M. avium and M. avium paratuberculosis and not the tuberculous complex (MTC). Until a decade ago, most cases of mycobacteriosis in birds were caused by mycobacteria of the MAI complex. However, nowadays a great number of different mycobacteria species have been identified from other avian species using modern laboratory technologies. So far only M. avium has been isolated from raptors (TELL et al. 2001). M. avium paratuberculosis, the causative agent of Johne’s disease has not been reported to occur in raptors, but in other avians (TELL et al. 2001). However, we isolated M. avium paratuberculosis from a saker falcon. Johne’s disease is very common in small ruminants in the UAE. It is therefore believed that feeding contaminated mutton to falcons may cause infection. Mycobacteriosis is common in free-living birds of prey as well as in raptors from zoological collections and those used for falconry (LUMEIJ et al. 1980). According to various authors mycobacteriosis accounts for 1 to 30% of the cases examined at postmortem. The highest incidence is from zoological collections. Compared to FABIAN and VETESI (1980) who reported 10% Falconiformes infected with mycobacteria in the Budapest Zoo between 1971 and 1978, the incidence of 1.9% in the Middle East hunting falcons is low. However, an increasing number of cases were observed during the last years, culminating in 8 cases in 2003. The clinical signs (particularly weight loss, carpal bursitis, palpable subcutaneous nodules) described in this case report are typical for this disease in falcons (HEIDENREICH 1995). The haematology findings (leucocytosis, monocytosis, anemia) in the clinical cases described in this paper are typical of mycobacteriosis (FUDGE and JOSEPH 2000), although aspergillosis and chlamydophilosis would need to be considered as possible differential diagnoses. HEIDENREICH (1995) states that the usual route of infection is oral and therefore the most common lesions in falcons are found in the gastrointestinal tract. However, other authors (TELL et al 2001) observed characteristic features in the liver spleen and intestines although any tissue may be affected. Our investigations clearly showed that the liver (17 x) was the prime organ, followed by spleen (14 x), peritoneum (7 x), skin (5 x) and lung (4 x). LUMEIJ et al. (1982) described tuberculous meningitis and encephalitis as well as knee joint involvement in raptors. Raptors seem to be more susceptible to mycobacterial bone infections than other avians. In buzzards (Buteo buteo) mycobacteria were isolated from bumble foot cases (TELL et al. 2001). It is worth noting that in our study mainly saker falcons (9 x) and saker hybrids (4 x) were infected with mycobacteria representing more than 50% of the cases. Three cases were seen in gyr falcons as well as in gyr hybrids and peregrines. In two cases, the species was not reported. 103 Due to the variation of the clinical appearance, avian mycobacteriosis is often not recognized on initial presentation and therefore remains challenging to diagnose in live birds. In the clinical cases described in this report other conditions, importantly chlamydophilosis and aspergillosis were diagnosed concurrently. The presence of other diseases can complicate the diagnosis of mycobacteriosis. Diagnosis is generally performed by ZN staining. To date, very little research has been conducted to develop appropriate tests for diagnosing mycobacteriosis in falcons. Although some diagnostic methods are considered more reliable than others, a direct comparison of diagnostic assays on a great number of samples is still lacking. In view of the zoonotic capacities of this infection and the close relationship between Arab falconers and their birds, therapy is not recommended by the authors. Other falcons that have been in close contact with affected birds should also be investigated and may need to be quarantined. 5 CITATION INDEX CHRISTOPHE C, VANNUFFEL P, BLONDEEL N, et al. Duplex PCR for differential identification of Mycobacterium bovis, M. avium, and M. avium subsp. paratuberculosis in formalin-fixed paraffin-embedded tissues from cattle. J Clin Microbiol 2000; 8: 3048 – 2054. 2. FABIAN L and VETESI F. Analyse der Vogelverluste (1971 – 1978) im Zoo Budapest. Verh. Erkrankungen der Zootiere 1980; 22: 215 – 221. 3. HEIDENREICH M. Birds of prey. Medicine and management. Blackwell Science. 1995; 117 – 120. 4. GRANGE JM, YATES MD and BROUGHTON E. A review: the avian tubercle bacillus and its relatives J Appl Bacteriol 1990; 68: 411 – 431. 5. LACHNIK J, ACKERMANN B, BOHRSSEN A, et al. Rapid cycle PCR and fluorimetry for detection of mycobacteria. J Clinical Microbiol 2002; 40: 3364 –3373. 6. LUMEIJ JT, DORRESTEIN GM and STAM JWE. Recent advances in the study of raptor diseases. Observations in tuberculosis in raptors. Proc. int. symposium on diseases of birds of prey, 1st – 3rd July, London, 1980. 7. TELL LA, WOODS L and CROMIE RL. Mycobacteriosis in birds. Rev sci tech off int Epiz 2001; 20: 180 – 203. 8. TELL LA, FOLEY J, NEEDHAM ML and WALKER RL. Diagnosis of Avian Mycobacteriosis: Comparison of culture, acid-fast stains, and PCR for the identification of Mycobacterium avium in experimentally inoculated Japanese quail (Coturnix coturnix japonica). Avian Dis 2003; 47: 444-452. 9. VAN DER HEYDEN N. Clinical manifestations of mycobacteriosis in pet birds. Semin. avian exot Pet Med 1997; 6: 18 – 24. 10. VITALE F, CAPRA J, MAXIA L, et al. Detection of Mycobacterium tuberculosis complex in cattle by PCR using milk, lymph node aspirates, and nasal swabs. J Clin Microbiol 1998; 36: 1050 – 1055. 1. 104 6 ACKNOWLEDGEMENTS The authors are grateful to Dr. P. McKinney, Dr. E. Pesci, Dr. A. Jawad, Dr. J.D. Remple and Dr. J. Samour for sending the samples. Furthermore we thank Mr. Chellappan Vishwanathan for his excellent histological slides. AUTHORS ADDRESS U. Wernery Central Veterinary Research Laboratory, PO Box 597, Dubai, United Arab Emirates. Email:
[email protected] 105 This bacterium has the faculty to produce extremely resistant spores in the environment: what are today the risks to see appearing such an outbreak? The authors present the epidemiological characteristics of the disease and the situation in the Loire Atlantique (France). the epizooty were due to type C toxin. In this paper. The Grand Lieu Lake is a wetland of international importance. In the case of Grand Lieu Lake. Marion L2 PhD. PhD. characterised by a limb paralysis. so called avian botulism. a major wetland in France. we present the results of a study aiming to understand the epidemiology of botulism in Grand Lieu Lake and to make management and prevention propositions. located near the Loire estuary in France.Management ABSTRACT Botulism is defined as a nervous affection. An epizooty in summer 1995 provoked the death of several thousands of birds among which. 2 THE DISEASE The toxin of C. This disease is a typically multifactor disease: it appears when environmental conditions allow the toxin production. 106 . These conditions are mainly anaerobic conditions. France EPIDEMIOLOGY OF AVIUM BOTULISM IN FRANCE : EXAMPLE OF GRAND LIEU LAKE Le Dréan-Quenec’hdu S1 DMV.Epidemiology . 1 INTRODUCTION The botulism is a nervous disease. after a brief review on the disease and the local context. characterized by flasks paralysis and induced by the ingestion of the toxin of Clostridium botulinum. this disease is due to a neurotoxin. birds of patrimonial importance.Waterbirds . Popoff M3 DVM PhD KEYWORDS Botulism . produced by Clostridium botulinum. They present the results of a study aiming at a better understanding and managing of the development of epizooty on a natural reserve. such as spoonbill or great egret.Eco-Ouest. botulinum provokes the disease: the presence of both bacteria and favoured conditions for toxin production are needed to trigger of the disease. The toxin production is due to local parameters. CATO et al. redox negative. bacterial growing and toxin production: there a lot of organic matter. to swim and can drown up. high temperatures. It’s classically divided into 7 toxinotypes. and then may amplify the phenomena. there are not specific lesions. sometime haemorrhagic. more than 174 species of bird species. concentrated in a very high way the toxin. In the “sludge bed” theory. The bacteria and its toxin C. The neurotoxins inhibit the neurotransmitters transmission. the neck and the eyelids. The paralysis seems to begin on the legs. 2. There are several theories on the needed conditions to have an outbreak. 1986). the outbreak is in relation with the environment conditions. in relation with antigenic specificities of the toxin (WOBESER 1981. anaerobic bacterium that forms dormant spores. When the muscles of neck and head are reached.2. for wild birds. such as flies’ larvae. then the wings. The resistance of the spore explains the persistence of the disease in a given environment. botulinum is a soil bacterium. The presence of mammalians or birds corpses is very often the departure point of the outbreak because: Clostridium botulinum is normally present in the intestinal microflore Corpses are suitable environment for bacterial growing and toxin production Scavenger invertebrates. In the microenvironment theory. 2. 107 . Spasmodically movements of eyelids may be seen. LAMARQUE 1995). particularly for taking flight and landing. Apparition of botulism epizooty should be interpreted as an indicator of environmental disequilibria: the massive death of waterbirds is often in relation with pollution of the environment (MOUTOU 1993. For birds there may be enteritis lesions. that is to say type C botulism. Epidemiological aspects C. The disease The disease is characterised by limb paralysis. Concerning avian botulism. favourable to the spore germination. the head of the birds stays either on the ground or on the bird back: the disease has been for a long time called “limberneck”.1.2.3. on the neuromuscular junction: the muscular contractions are blocked. A B C D E F G. The death appears after respiratory paralysis. Botulinic birds are enable to walk. Wild birds show flight difficulties. They let them easily catch (POUANT 1997). theses larvae ma be eaten by birds. and toxin producing bacterium. Generally. pH between 7. botulinum is a bacterium normally present in the sediment.5 and 9. the death is often due to predation or the drowning for aquatic birds. form 22 families have been implicated in an epizooty. anoxia. Heavy and green diarrhoea is often seen. there is a relatively independence of the environment conditions. Since 1994. There were not significant differences between positive and negative stations for any physicochemical parameters. such as raptors. 102 samples of mud have been made. (1995. but all the families were affected. 2. pH. Results The results of 2001 study shown that 17. pH. a forth sites “La Capitaine” appeared to be a “risk” site. aiming to map the bacteria presence. There were 3 sites of risk. conditions of oxygen content. redox.3 CASE STUDY : LAKE OF GRAND LIEU IN LORE ATLANTIQUE (FRANCE) 3. 73 % of the diseased / death birds were ducks. we measured also water level. redox and temperature suitable for toxin production. Samples of mud were sent to Pasteur laboratory in Paris. organic matter in the mud. 24 French departments have been reached by botulism outbreaks. The same parameters were measured. with a speedboat. the physicochemical conditions in June 2001 did not allow toxin production: any positive station. even those uncommonly affected. Materials and methods In June 2001. each day. 1995 is the year of the first heavy outbreak described: 36 species have been contaminated. 1996). except for depth oxygen parameters: 108 . However. the water levels were increased on the whole lake and management measures were recommended (for example clean up of the streams). allow probably limiting the extension of the mortality. during the botulism epizooty. surface and depth parameters. several were made on the same location than in 2001. showed at the same time. in order to search for Clostridium botulinum and its type C and D toxin.6% of the samples were positive for type C or/ and type D. 48% of the sampled stations were positive but the sample was not made according random way. Summer 2003 was very hot summer. oxygen in surface water. but mainly mallards (see BORET and REEBER 2003). Ruby and La Morne. 3. Analyses were performed by mouse test after and before enrichment and by double PCR as described by FACH et al. 3. more than 800 corpses were collected. according to the presence of bacteria: Sénaigerie. in depth water and in mud. The results confirmed the presence of 3 “risk” sites: Moreover. the GPS position was measured. In summer 2003. water and mud temperature. Another outbreak has occurred in Grand Lieu Lake and in several areas in France. a first study has been conducted. In Grand Lieu Lake. without to count the passerine birds. 3. In Grand Lieu Lake. Following this outbreak. The systematic collect of the corpse. according to meteorological conditions and local context. On each point. 1 Local context Mass mortality on waterbirds may occur in several areas in France. There were not significant differences between positive and negative station according to water level. except one. MARION (1995) estimated that 90% of the Marsh Harrier died. The distribution of positive stations was not due to chance. amongst which. 34 samples have been made. In summer 2003. but not in the whole lake. despite the absence of bacteria in 2001.118.the oxygen content of depth water is significantly lower for negative station (mean = 0. 4 MANAGEMENT AND PREVENTION OF BOTULISM EPIZOOTY 4. Management during the epizooty In case of outbreak. It is possible to identify “risk sites” but further analyses are needed to improve these sites. Morne…). by opposition to the installation of the paralysis which is ascending.e the bacteria in the sludge bed theory are present in the lake.1. they contaminated this site. Other vaccines were tested with more or less of success to birds. the mud temperature was higher than 23 °C for all positive station except one. Moreover. These conditions lead the high mortality observed. For some site. it is fundamental to proceed to a systematic collect of corpses to avoid the enrichment of the environment. p = 0. When physicochemical conditions of environment are suitable for toxin producing AND birds are present in high density (such as during moulting period).873.382. possibly antibiotic. Treatment of the birds In benign cases.5) and oxygen content is very low (lower than 1. standard deviation negative station = 0. for all the positive station except one. notably speedboats that allow reaching the zones of weak depths. However. the physicochemical conditions were suitable to toxin production and then for the outbreak.045). negative station standard deviation positive station = 2. t = -2. the microenvironment theory explains the amplification of the outbreak. the bird can recover in 2 in 3 days if it is removed from his environment: the recovery is downward. were birds stays in very large flocks. We can then assume that: the conditions i. in June. like “La Capitaine”.127. 4. vitamins. mainly during moulting season. In 2003.2. When the contaminated birds died on the feeding place. such as “La Capitaine”. an outbreak may occur like in summer 2003. This was made during summer 2003 on the Grand Lieu Lake but it is essential to finalize a protocol of reproducible collect: it will also allow following the mortality from one year 109 . such as in pounds (Sénaigerie.5). pH is suitable for toxin production (lower than 7. mean positive station = 1. The presence of dead birds on “La Capitaine”. There is no immunity post-infection because the toxic dose is lower than the immunizing dose: the immunization against the disease can thus obtain only with the use of specific anatoxins (inactivated toxin) (POUANT 1997). MARTINEZ and WOBESER (1999) showed for example that the administration in ducks mallards of a vaccine on base of inactivated Clostridium botulinum (usually used in minks farms) can protect birds and suggested to administer it to birds in the same time as the treatment. We shall administer a treatment of support: rehydratation.864. may suggest that the birds did not primarily contaminated in this site: the contamination may be done on the roosting place. This collect has to appeal to appropriate means. to clean out ponds. 78: 2 . Investigation of animal botulism outbreak by PCR and standard methods.23. when it is possible. In term of water level. (eds): Bergey’s Manual of Systematic Bacteriology.3. 5. Abiotic predictors of avian botulism outbreaks in Utah. 154: 18 . In: SNEATH PHA. FACH P.to the next. in particular if sediments remain rich in water. GIBERT M. 13: 279 .1161. sites into small subunits. MAIR NS. MARION L. GIBERT M. outside the periods of strong temperature. GEORGE WL and FINEGOLD SM. 1995. E. Genus Clostridium Prazmowski 1880. 209: 17-24. so as to be able to go up or lower quickly the water level on these micro-sites according to the outbreak risk. SHARPE ME and HOLT JG. 4. when it is possible. that is to say in winter. We can recommend thus on the contrary maintaining water on a sufficient level and especially stable from the beginning of the warm season. Prevention of the epizooties Concerning the organic matter content. If the dryness is not complete. Le botulisme à Grand Lieu : une catastrophe écologique majeure. BORET P and REEBER S. vol 2. Corpses are cremated. FACH P. CATO EP. Le Courrier de la Nature. BARRAS and KADLEC (2000) recommended to divide. and evaluation in food samples.729. GRIFFAIS R and POPOFF MR. Appl Environ Microbiol 1995. 1157 . 61: 389 . Note d’information de l’ONC. Birds alive are directed to a wildlife centre. LAMARQUE F. Le Courrier de la Nature 2003. we can limit the vegetable production. 110 . Wildlife Society Bulletin. B. 8. 28: 724 . 5 CITATION INDEX 1. Lac de Grand Lieu: résultats des operations de ramassage des oiseaux atteints de botulisme sur la reserve naturelle en 2003.392. PCR and gene probe identification of botulinum neurotoxin A. Baltimore: Williams & Wilkins 1986. LE DREAN-QUENEC’HDU S. and possibly when banks are weakly tilted to modify their hillside. On the same way if strong rains arise in spring. Botulisme. 1995. GRIFFAIS R. 6. F. 3. 7. we have on the contrary the risk of favouring the toxin production by allowing the increase of sediment temperature.285. Etude du risque sanitaire représenté par le botulisme sur le lac de Grand Lieu. If it is possible (it is not the case of the Grand Lieu Lake) it is necessary to go to raise brutally the levels of water in order to lower the temperature of the sediment and possibly make inaccessible the contaminated sediments. It is also necessary to identify and to fight the other causes of mortality. 4. et al. Mortalités massives. The wildlife centre must be equipped with swimming pools as it is the case of the wildlife centre of Nantes. and G-producing Clostridium spp. 2. Rapport non publié pour la SNPN. hydraulically independent. BARRAS SC and KADLEC JA.The preservation of a predatory and scavengers population can help to limit the number of corpse and thus the risk of outbreak. FEMS Immunol Med Microbiol 1996. 2000. Au service de SAGIR. 2001.3. the most radical solution is the complete dryness of the site. France Email: sldq@club-internet. 35520 Melesse. Les animaux sauvages sentinelles de l’environnement. The rest of the references are available from the author AUTHORS ADRESS Le Dréan-Quenec’hdu S. Immunization of ducks for type C botulism. 24: 667 . Le Point Vétérinaire. 35: 710 . WOBESER G.9. 1993.672. J Wildl Dis 1999. PhD Eco-Ouest 3 rue de la Janaie. 10. MOUTOU F.fr 111 . DVM.715. MARTINEZ R. Flies collected off sick or dead owls tested positive for WNV RNA.Owl . A WEST NILE VIRUS OUTBREAK IN NORTH AMERICAN OWLS A. While in humans and horses only a minority of WNVinfected individuals develop severe clinical disease.Icosta Americana . Lindsay2. The Ontario Veterinary College. 2003). Hunter1. mammals. 2001. Canada. Canadian Science Center for Human and Animal Health. I. MSc. West Nile Virus (WNV) has emerged as a health risk to humans and has been associated with morbidity and mortality of a wide variety of North American birds. 1 INTRODUCTION Since its initial detection in the New York City area in 1999. ABSTRACT Between July and September 2002.Epidemiology . MILLER et al. Health Canada2. Gancz1. KEYWORDS West Nile virus . Most of the survivors (75.K. A.Ontario . 2003).8%) were seropositive for WNV. National Microbiology Laboratory. DVM. Barker1. some bird species appear to be highly susceptible to this virus (EIDSON et al.Deptartment of Pathobiology1. PhD. Canada. The Owl Foundation3. DVM.Hippoboscida . the factors that make some species highly susceptible to WNV remain unknown. 112 .Y. Dibernardo2. Vineland. and reptiles (McLEAN et al. McKeever3. and B. Family Hippoboscidae) infestation. PhD. Peak mortality occurred in mid August and corresponded with mortality of wild corvids in the surrounding region. an outbreak of West Nile Virus (WNV) caused high mortality of captive owls at The Owl Foundation. K. Species with northern breeding range and of medium/large body-size were at significantly higher risk of exposure to WNV. 2002. R. Ontario. Species with northern breeding range were at significantly higher risk of WNVrelated mortality. DVM. and died earlier during the outbreak. The outbreak occurred in the midst of a louse fly (Icosta americana. University of Guelph.Canada. To date. KOMAR et al. MSc. this is the largest WNV outbreak in a captive wildlife collection in North America. as some closely related species (e. were found to be WNV positive based on reverse transcription polymerase chain reaction (RT-PCR). between July 26 and September 28. body size. Vaccination of the remaining birds with a killed WNV vaccine was attempted at the face of the outbreak. From July to September 2002. The organisation maintains detailed records of all birds in the facility. however. The objective of this study was to test the effect of outdoor housing. records and observations The Owl Foundation specializes in breeding and rehabilitating North American owls. 108/245 (44%) owls died. tissue samples from eight owls. These outbreaks are of special interest as they offer a unique opportunity to study the impact and epidemiology of WNV infection in multiple species under quasi-natural conditions. This data was gathered by the Niagara Region Health Unit between May 14 and October 12. On August 16. 2003). The role of louse flies in transmitting WNV was also examined. Ontario.g. 79°20’ W) has approximately 3340 M2 of specially designed outdoor cages and a few indoor cages. 113 . Using a 20G spinal needle. 2002. This data was used in the analysis of this outbreak.Little is known about the effect of the host’s taxonomic. American crows (Corvus brachyrhynchos) and fish crows (Corvus ossifragus) show different susceptibility (KOMAR et al. Necropsy findings included marked hepatomegaly. taxonomy (at the sub-family level) and geographical range on exposure to WNV and on WNV-related mortality. Its facility in the Niagara region (Vineland. lung. Taxonomy alone offers limited help in predicting susceptibility to WNV. 2002. high mortality occurred in captive owls kept at The Owl Foundation (TOF). and the first one in Canada. 43°10’ N. At the time.. For each bird. many of the birds were infested with adult haematophagous louse flies (order Diptera. Ontario. Samples of tissue. Data on dead Corvidae sightings in the Niagara region was obtained from the Canadian Cooperative Wildlife Health Center national WNV surveillance database. including the initial three. blood and flies Complete diagnostic necropsies were performed at the Ontario Veterinary College on 94 owls and one falcon that died at TOF between April 15 and December 25. Vineland. splenomegaly and cerebral haemorrhage. Initially the deaths were attributed to this infestation and only on August 9 did the authors examine three dead owls. geographic and demographic background on its susceptibility to WNV. 2 MATERIALS AND METHODS Study site. samples (approximately 1mm3 in size) of brain. Canada. small core samples of brain. age. family Hippoboscidae). spleen and kidney were pooled and tested for WNV RNA by real time RT-PCR. kidney and liver were collected from three additional carcasses that were not available for necropsy. liver. To the authors’ knowledge. Homogenates prepared from whole louse flies were tested using the same procedures described for owl tissues. A selected sample of flies. salivary glands. 10 and 1 PFU and a negative control (media only). Positive controls consisted of three ten-fold dilutions of Egypt 101 (E101) strain of WNV. Species were considered „Northern“ if their reported natural breeding range was mainly north to latitude 48°N or „Other“ if it was not so (MARKS et al. 2001). Three sets of negative (water) controls were used. Study Population The study population (SP) included 245 birds of 17 species that were present at TOF on the first day of the WNV outbreak (based on RT-PCR results). pupae were collected of the bottom of the cages. A subset of 20 plasma samples representing 8 species of owls and 4 species of raptors was tested both by ELISA and by plaque reduction neutralisation tests (PRNT). were dissected and specific body parts (e.During the outbreak louse flies were collected at TOF off sick. abdomen and thorax) tested for WNV by RT-PCR and immunohistochemical (IHC) staining. Statistical analysis Logistic regression was used to test the effect of outdoor housing. Samples that had CT ≤ 37 with both primer sets were considered positive. some had spent many years at TOF while others were recent additions or hatchlings of the year. 2002). Plasma was then separated by centrifugation and frozen at -70°C until analysed. age and species body size on exposure to WNV and on WNV-related mortality (among exposed birds). University of Northern Iowa). Adult flies and pupae were pooled by cage and tested for WNV RNA by real time RT-PCR. Most were permanently disabled birds of wild origin. 2000). taxonomy (at the subfamily level) geographical range. hind gut. collected off WNV-positive birds. Ten birds were housed indoors while 235 were kept outdoors. The assay was performed as described previously (KOMAR et al.g. A serological survey of all outbreak survivors was conducted. including a barred owl (Strix varia) (EBEL et al. The controls used were back titrations of WNV (NY strain) at 2. Positive samples underwent a second RNA extraction and were then tested by both the 3‘NC and ENV primer sets (LANCIOTTI et al. The assay was performed as previously described with slight modifications. 2000). two during the extraction procedure and one during amplification. Species were further classified as 114 . 1999). Blood samples placed in heparinised tubes. Serology We used an enzyme-linked immunosorbent assay (ELISA) shown to detect avian antiWNV immunoglobulin G (IgG) in 23 avian species of 12 orders. Real-time (TaqMan) RT-PCR Real-time RT-PCR assay was used to detect WNV RNA as previously described (LANCIOTTI et al. Six adult flies were submitted for species identification (courtesy of Nixon Wilson.5x107 PFU/ml diluted to 100. Extracts were screened using the generic 3‘NC primer set. In addition. dead and asymptomatic birds. The two age groups compared were <1year and >1year. Size and subfamily were not found to be significant risk factors for WNV-related mortality.79-74. Birds older than 1 year were also more likely to die of WNV (P<0. Agreement between ELISA and PRNT results was good (Kappa = 0. Large or medium birds also died earlier during the outbreak (mean 21. 1999).05).557). NC. n=8). Subfamily and age were not significantly associated with the date of death.e.01). Species in the low MR group have intermediate or very broad range of distribution (Table 1). All species in the high MR group have northern native range. 115 .5 days. 95% CI 0. Northern species died earlier during the outbreak (mean 23. CA) was used to test for agreement between ELISA and PRNT results.2 days.58-1. SAS Institute Inc. 79/85 (92. Of these. n=77) compared to small species (mean 33. n=93) compared to other species (mean 35.51.46-9.82 (3..9 ±11. A general linear model was used to test the effect of the same factors on the date of death (i. Regression procedures were performed using the SAS software (Version 8.05) with odds ratio (OR)=52. 250-500g.2. Three distinct groups were seen: high MR (90-100%). The first five cases occurred over a period of 11 days in five different cage complexes scattered throughout the facility. Species northern breeding range and large/medium body size were significant risk factors for exposure to WNV (P<0. low MR (MR< 16. respectively. Mortality rates (MR) differed greatly between species.857.13). and no mortality (MR = 0%). Among exposed birds.8%) were seropositive based on ELISA.3 %.26. Inc. USA). Being kept outdoors during the outbreak was found to be a highly significant risk factor (P <0.84) and OR=16. while species that had MR of 0% have relatively southern range.05) with OR=4.0001) for WNV-related mortality.67) respectively.13-881. medium or large if their average body weight was <250g.9 ±8. Age and taxonomy (at the subfamily level) did not significant influence exposure.9 %) birds that died tested positive for WNV. The patterns of daily mortality at TOF and dead corvid sightings in the Niagara region during this period show close resemblance. kept outdoors during the entire outbreak and not vaccinated against WNV.7%). A total of 91 outbreak survivors. or >500g respectively (MARKS et al. were tested for anti-WNV IgG. n=24) (P<0. The Kappa statistic (Quickcalcs. Cary. the first and last cases of WNV-related mortality at TOF were on July 26 and September 28.0 ±12.small. the number of days to death from the index case).8 ±9. Within the outbreak period.56 (3.9 days. 3 RESULTS Based on RT-PCR results. 69 (75.8 days.0001) with OR=1507 (85. northern range was a highly significant risk factor for WNVrelated mortality (P<0.62). GraphPad Software. Geographical range was significantly associated with the date of death (P<0. The overall exposure rate (ER) was 84.87 (2. Taxonomic affiliation at the subfamily level did not significantly affect MR. as for other pathogens. this would explain the overall high ER seen at TOF in light of the wide spread louse fly infestation. and the host’s body size may be an important determinant of vector attraction. Immunity to WNV. Further data. The similarity between the mortality pattern at TOF and that of dead corvids in the Niagara region and the fact that the initial cases occurred in several cages scattered throughout TOF. 2002).g. It is possible that northern species have a less effective cell-mediated immune response to pathogens. This link is intriguing. T-cell mediated immunity is essential for fighting fungal pathogens (LEHMANN 1985) and is also believed to play an important role in the immune response against flaviviruses (ANRAKU et al.g. WNV RNA was detected by real-time RT-PCR in 16/18 (88.The six louse flies examined were identified as Icosta americana (Order Diptera. As none of the birds kept indoors were affected.9%) flies collected off dead or sick owls during the outbreak. would have been less likely to have acquired immunity. young birds.3%). if this was the case. Looking at the species specific MR. it would be expected that all owls that spent years at TOF would have similar susceptibility. Innate immunity could potentially have evolved through selection if some of the species have coexisted over long periods of time with agents similar to WNV. Louis Encephalitis Virus) 116 . Susceptibility to WNV-related mortality crossed taxonomic lines and was strongly related to geographical range. the major route of WNV transmission likely was vector borne. Given the high overall ER. this finding suggests marked differences in species susceptibility to the virus. can be either innate or acquired. Our data shows the opposite. Northern owls may have little or no exposure to flaviviruses (e. Family Hippoboscidae).) has been documented. including IHC results will be presented as it becomes available. 4 DISCUSSION West Nile virus RNA was present in the vast majority of owls that died at TOF during the outbreak and antibodies against WNV were detected in most outbreak survivors (overall ER 84. suggests that the outbreak was part of a regional WNV activity. Northern geographical range and large/medium body-size were significant risk factors for exposure to WNV. which are scarce or nonexistent in their natural environment. St. Furthermore. especially since a similar relation with regards to susceptibility to other pathogens (e. Five adult flies that were collected off healthy appearing birds were all WNV negative. Aspergillus sp. If this parasite can transmit WNV. If immunity in this case was acquired. regardless of species. Northern species due to their thicker feathering may attract more feather-dwelling arthropods. Birds that attract more vectors have a higher risk of exposure to WNV. it is clear that the distribution of WNV-related mortality was uneven. WNV RNA was detectable in Icosta americana louse flies collected off dead or sick birds. Heather White for technical assistance. Experimental infection of North American birds with the New York 1999 strain of West Nile virus. LANGEVIN S. . Factors such as size and age may (to a lesser magnitude) affect exposure and susceptibility respectively. however this requires further examination. 7: 615 .gov/ncidod/EID/vol9no3/02-0628. Crow deaths as a sentinel surveillance system for West Nile virus in the northeastern United States. Detection by enzyme-linked immunosorbent assay of antibodies to West Nile virus in birds. Emerg Infect Dis 2002. 7: 621 . et al. et al.cdc.3799. 1999. HARVEY TJ. As WNV continues to spread. J Virol 2002. KOMAR N. J Clin Microbiol 2000. ANRAKU I.and may therefore be particularly susceptible to WNV.gov/ncidod/EID/vol8no9/020152. Available from: URL: http://www. Rapid detection of west nile virus from human clinical specimens. 5 CONCLUSION There is a strong link between geographic range and susceptibility to WNV in North American owls. Further investigation is needed in order to answer this question. Jody Crossingham. and Hillary Voet for assistance in statistical analysis. 117 . KERST AJ.625. Available from: URL: http://www.620. or a combination of the above. shorter disease course. 6 ACKNOWLEDGEMENTS The authors thank Annick Gionet. and Amanda Low for their assistance in data collection. et al.htm EIDSON M. common avian haematophagous parasites. DUPUIS AP II. Serologic evidence for West Nile virus infection in birds in the New York City vicinity during an outbreak in 1999. et al. Emerg Infect Dis 2003.htm LANCIOTTI RS. 2. The Rathlyn Foundation for the stipend that made this study possible. 7 CITATION INDEX 1. Chris Enright. Dale A. BURNS JE. field-collected mosquitoes.4071. HINTEN S. West Nile Virus Avian Mortality Surveillance Group. 38: 4066 . Douglas Hodgins. and avian samples by a TaqMan reverse transcriptase-PCR assay. shorter incubation. free ranging populations of susceptible owl species may be significantly affected.cdc. 5. Nathalie Lemieux for help in documentation. et al. larger infective dose. SORHAGE F. et al. Smith for critical reading of the manuscript. Emerg Infect Dis 2001. Kunjin virus replicon vaccine vectors induce protective CD8+ T-cell immunity. EBEL GD. 76: 3791 . Emerg Infect Dis 2001. may play a role in transmitting WNV. 4. Louse flies. This relationship crosses taxonomic lines at the subfamily level and may be related to differential immune competence. This could be a result of infection at earlier date. NICHOLAS D. Kara Kristjanson. 6. PANELLA NA. 3. Northern distribution and large/medium body-size were significantly associated with earlier death during the outbreak. NASCI RS. KOMAR N. KOMAR N. LINEDALE R. 10. Vol. LEHMANN PF. UBICO SR. West Nile virus in farmed alligators. et al. Emerg Infect Dis 2003. CANADA. editors. Immunology of fungal infections in animals. 267: 271 . 10: 33 . DVM.308. 9. ON. 34 . MCLEAN RG. The Ontario Veterinary College.cdc. BOURNE D.gov/ ncidod/EID/vol9no7/03-0085. of Pathobiology. Vet Immunol Immunopathol 1985. KOMAR N. In: Del Hoyo J. Elliott A. Curr Top Microbiol Immunol 2002. University of Guelph. Gancz.ca 118 . and Sargatal J. MAUEL MJ. N1G 2W1 Email: agancz@uoguelph. CANNINGS RJ. Barcelona: Lynx Edicions 1999. MIKKOLA H.htm AUTHORS ADRESS Ady Y. MSc. MARKS JS. MILLER DM.243. BALDWIN C. Available from: URL: http://www. 8. Family Strigidae (Typical owls). Handbook of the birds of the world. 5 Barn-owls to Hummingbirds. et al. Dept.7. West Nile virus in livestock and wildlife.69. Guelph. 0 43. 1 Based on real-time RT-PCR.Table 1.0 0.1 7.7 12.5 11.0 0.0 100.3 90.September 28.0 119 .9 92. Mortality at The Owl Foundation property during a West Nile virus outbreak .5 0. 2002) Species Snowy owl (Bubo scandiacus) Northern hawk owl (Surnia ulula) Great gray owl (Strix nebulosa) Boreal owl (Aegolius funereus) Northern saw-whet owl (Aegolius acadicus) Northern pygmy owl (Glaucidium gnoma) Short-eared owl (Asio flammeus) Flammulated owl (Otus flammeolus) Long-eared owl (Asio otus) Great horned owl (Bubo virginianus) Barn owl (Tyto alba) Barred owl (Strix varia) Burrowing owl (Athene cunicularia) Eastern screech owl (Megascops asio) Elf owl (Micrathene whitneyi) Spotted owl (Strix occidentalis) Tawny owl (Strix aluco) American kestrel (Falco sparverius) Peregrine falcon (Falco peregrinus) Total: MR = Mortality rate (calculated when n>6).(July 26 .7 4.3 16.0 91. At Risk 20 26 27 11 13 6 16 9 13 22 10 8 10 36 1 1 2 2 2 235 WNV-related1 MR % 100.0 0. sedentary or migratory species. D. Mortamais. It was isolated for the first time in 1937 in Uganda. Augé.Epidemiosurveillance . The avifauna is the virus’ main reservoir. no abnormal mortality due to the WNV virus was reported. seven human and four equine cases were observed in the Var district. In 2001 and 2002. belonging to the Flaviviridae family like the yellow fever virus. Hars.Office National de la Chasse et de la Faune Sauvage Unité Sanitaire de la Faune. and humans and horses sensitive hosts (accidental victims). the French game and wildlife agency (ONCFS) conducted a serological study of 5 bird species in the Camargue.Avian disease . The monitored area was extended in 2004. France SURVEILLANCE OF WEST NILE VIRUS IN THE AVIFAUNA OF SOUTHERN FRANCE J. 120 . as an early warning system. S. Schuffenecker. Zeller KEYWORDS West Nile virus . H. whereby low seroprevalences were detected in mallards (8%) and magpies (22%). (Corvids. In 2000. West of the Nile (after which it has been named) in the serum of a young woman with a febrile syndrome. G. mallards and chickens located in 30 sites distributed along the coastal region.France ABSTRACT West Nile fever is a mosquito-borne viral disease for which wild birds are amplifying hosts. Balanca. Chavernac. These results show the interest of monitoring WNV circulation in birds. In 2004. there is an epidemiosurveillance program for avifauna. It reappeared in 2000 and 2004 in the same region involving only horses. Zientara. Many wild. Pradel. only 2 seroconversions were detected in sentinel birds suggesting a very low circulation of WNV. Since 2001. J. It is based on the detection of abnormal mortality in wild birds and on a serological monitoring of sentinel birds. West Nile virus (WNV) was recognized for the first time in southern France (Camargue) in 1962-1965. M. Among the avifauna. since birds are the hosts that will amplify the viral circulation. 1 INTRODUCTION The West Nile virus is an arbovirus (arthropod borne virus). 200 km East from the Camargue region. I. In 2003. P. before the occurrence of equine cases. with the help of several seroconversions the presence of the virus was revealed in sentinel birds. Passerines) but also domestic ones (Turkey. 21 deaths) (ZIENTARA et al. were American crows (Corvus brachyrhynchos) and blue jays (Cyanocitta cristata) (ZELLER and SCHUFFENECKER 2004). 2001). between the month of August and the month of November the horse population was contaminated (76 confirmed cases. this is a mosquito of the Culex genus (C. nor transmitted. some vectors may bite horses as well as humans and transmit the virus to them. PLATONOV et al. the mosquito will feed on the blood of its host. 2001. neurological manifestations with high mortality rates have been observed among pigeons in Egypt (TAYLOR et al. In general. Gard and Bouches-du-Rhône carried antibodies (source 121 . no curative treatment nor vaccine is available against the West Nile virus. that contaminates itself when it bites an infected bird. and may go from a simple flu syndrome to encephalomyelitis with a high mortality rate. An exhaustive serological survey revealed that 8. is determined by the activity of the mosquito vectors. may lead to complications like encephalitis or aseptic meningitis that might be lethal. 2001).e. In some cases. Hérault. but the cases of mortality were mostly observed in corvids which. Rumania. After a phase of dormancy that lasted for more than 35 years. In Man like in birds. MURGUE et al. 2000) and Asia. France) (TSAI et al.000 horses were infected. but also in Europe (Portugal. 1956. 2000). West Nile fever is transmitted by a stinging arthropod vector. 4. In 2003. Waders. are generally considered to be “epidemiological culs de sacs”: i. the Middle East (WEINBERGER et al. The virus was isolated in 1964-65 in men and in mosquitoes of the Culex modestus species (JOUBERT et al. later on. through them the virus can neither be multiplied.862 human cases were described and 264 of the sick died (CDC 2003).133 horses that had been tested in the regions of the Camargue. Chicken) may host and transport the virus (ZELLER and MURGUE 2001). When. however.3% of the 5.Laridae. mainly among the older ones. where it had contaminated horses (500 clinical cases with a morbidity of 25% and a lethality of 10%) and humans (19 clinical cases. east of the city of Montpellier. 1970. the clinical signs of the disease are extremely variable. where. humans also may develop a pyretic syndrome which. in turn. The length of this cycle depends on the climate (temperature. it could transmit the virus to another sensitive host. 1953) and Corvids in the United States (STEELE et al. The occurrence of the disease has been described many times in Africa (MC INTOSH et al.500 died. Generally. modestus in Europe). When an intense bird-mosquito cycle is going on. 2000. In France. it suddenly reappeared in August 2000 in the area of the Petite Camargue. The virus multiplies in the arthropod and then reaches its salivary glands. Italy. However. 1998. WORK et al. 2000). 1971). In North America. Anatidae. HUBALEK and HALOUZKA 1999. In 2002. West Nile fever is usually subclinical. Today. 10 of which were severe). humidity) which. most often. birds are «asymptomatic carriers». Among horses. pipiens or C. 1976). 9.. The virus appeared for the first time in 1999 in the United States in the region of New York and circulates now throughout the whole North American continent (STEELE et al. CANTILE et al. West Nile virus was described for the first time in 1962-63 in the Grande Camargue area. out of which 4.156 clinical human cases out of which 284 cases of deaths were filed while 15. Goose. Raptors. Like any other arbovirus infection. Accidental hosts like horses and Man. BALANCA and HARS 2005). the West Nile virus was isolated in 234 bird species (HUBALEK 2000. is carried out in the infected area. In late autumn 2000. 2001. their identification and the virus isolation. This work was carried out thanks to an awareness campaign for the general public based on an important advertising campaign. Ministry of Agriculture) and the DGS (General Direction of Health Affairs. Office national de la chasse et de la faune sauvage. since not a single human case of meningitis-encephalitus was recorded and no abnormal or unusual mortality was observed in the wild and domestic birds (CHEVALIER et al. n = 81) were negative (HARS et al. 122 . 0. Gard et Bouches du Rhône). with an enhancement in the Camargue region and in the Mediterranean “départements”. and practising veterinarians. the Center for International Cooperation in Agronomic Research (Centre de coopération internationale en recherche agronomique pour le développement. a serological survey in five avian species was made in the outbreak area with the help of ELISA tests confirmed by seroneutralisation. and on virological analyses of all dead birds found by the members of the SAGIR network (the national network for the surveillance of the causes of mortality in wildlife). an entomological survey based on captured mosquitoes. As soon as the occurrences of human or equine cases are confirmed. a national surveillance programme for the West Nile virus has been implemented by the DGAl (General Food Administration. n = 117) and blackheaded gulls (Larus ridibundus. Birds tested positive for the West Nile virus were found in three species with low prevalences: in 8% of the mallards (Anas platyrhynchos. Ministry of Health).9% of yellowlegged gulls (Larus cachinans. This programme is based on the surveillance of the cases of human encephalitis in hospitals and equine encephalomyelitis occurring throughout France. n = 100). the départemental veterinary services. in collaboration with the National Reference Center for Arboviruses and Haemorrhagic Viral Diseases of the Pasteur Institut (CNR-IP). It seems that only equids were clinically infected by the virus. 2004) 2 MATERIAL AND METHODS Since 2001. the objective of the surveillance is to be able to rapidly detect any circulation of the West Nile virus in the Camargue so as to give an early warning before any clinical signs in equids or humans are revealed. all tested House sparrows (Passer domesticus. the départemental hunter federations. The implementation of this avian surveillance program is performed by the wildlife sanitary Unit of the French Hunting and Wildlife Agency (Unité sanitaire de la faune. In 2001. n = 18). Concerning avifauna.AFSSA Alfort). a large spread of posted public notices and a Freefone telephone number (Fig 1). surveillance was based on: � A «passive» system based on a closer surveillance of the cases of avifauna mortality in the départements of the Camargue region (Hérault. The research of the virus in the brain of dead birds is carried out by RT-PCR and viral isolation on a cell culture. 2002). However. 2002 and 2003. n = 114) and 22% of black-billed magpies (Pica pica. ONCFS). the départemental veterinary laboratories (LDAV). CIRAD-EMVT). and to take the appropriate measures to inform the public and prevent the disease. The first blood sample taken allowed to control whether a bird was serologically negative. The sera. 123 . an average number of 200 «calling ducks» (mallards breed by hunters and released on the ponds and in the marshes to attract the wild ducks) and domestic chicken. i. where no aviary West Nile surveillance had been set up.e.� an «active» system that is based on a serological survey of sentinel birds: i. Poster used for advertising campaign In 2004. collected and processed in the LDAV. Fig 1. the protocol was modified because of the emergence of several cases of human and equine encephalomyelitis in September 2003 in the Var département. An individual code number was attributed to each bird in order to monitor the changes in its serological status throughout the whole monitoring campaign. was then considered as an incontestable sign of its recent contact with the virus. were transmitted without delay to the CNR where they were treated by ELISA IgG with confirmation of positive results by sero neutralisation. blood samples were monthly taken (from June to November). distributed over 20 sites . the appearance of antibodies in an initial seronegative bird. A sero-conversion. There were 10 to 12 birds per site which were identified with two coloured rings on one of their legs.e. From each bird. 1. by all partners of the program.fr. It may be consulted online on an Internet site with protected access: http:// west-nile. Fig 2. Geographical distribution of sentinels birds sites in the South of France in 2004 The collection. and is located at. A leaflet to inform and incite the public to collect the dead birds has been distributed to the institutions or associations concerned by avifauna.� � a passive system has been applied in ten mediteranean départements . Bird mortality See table 1 for the list of wildfowl mortality cases that were registered in the area of surveillance through a toll-free telephone number and by the SAGIR network. A total number of 320 birds. 3 RESULTS 3. Aude and Var.cirad. distributed over 31 sites (Fig 2) was controlled every month. CIRADEMVT. treatment and circulation of all data collected through the surveillance of the avifauna is done with a data base that was set up by. an active system has been extended to three other départements: Pyrenées Orientales. 124 . Table 1: Reference table for the cases of avian mortality recorded in the area under surveillance between 2001 and 2004 Date July 2001 August 2001 August 2001 August 2001 July - August 2002 August 2002 August 2002 August 2002 August 2002 August 2002 August 2002 May 2003 August 2003 AugustSeptember 2003 August 2003 August 2003 August 2003 September 2003 October 2003 July 2004 August 2004 August 2004 September 2004 Species Waterfowl Wood pigeons Mallards Mallards Coots Waterfowl House sparrows Mallards Mallards Mallards, Garganeys, Teals Mallards Gulls, Waders Gulls Mallard Waterfowl House sparrow Mallards Pigeon Turtle doves Ducks DucksTurtle dove Partridge Pheasant Chicken Sheldduck Pigeons Place - Commune (département*) Several communes (13) Lansargues (30) Vauvert (30) Marignane (13) Several communes (13 and 30) Vauvert (30) Sète (34) Marsillargues (34) Fleury d’Aude (11) St Laurent d’Aigouze (30) St Nazaire de Pézan Arles (13) Villeneuve les Maguelones (34) Several communes (30 and 34) Montpellier (34) Carcassonne (11) Nîmes (30) Martigues (13) Carcassonne (11) Le Caylar (30) Nîmes (30) Draguignan (83) Montpellier (34) Carcassonne (11) Presumed (P) or confirmed(C) cause of mortality Botulism (C) ? Intoxication pesticides (C) Botulism (C) Botulism (C) ? ? Ulcerative enteritis (C) ? ? Paralysis (P) Botulism + diarrhea (C) Salmonellosis (C) Botulism (C) ? Intoxication (P) ? ? ? ? ? ? ? West Nile virological findings Negative Negative Negative Negative Negative Negative Negative Negative Negative Negative Negative Negative Negative Negative Negative Negative Negative Negative Negative Negative Negative Negative Négative * départements : 11 = Aude ; 13 = Bouches du Rhône, 30 = Gard ; 34 = Hérault ; 83 = Var 125 First of all, we can notice that the number of dead birds that was declared and for which an autopsy in the laboratory was made, is very small in spite of all the information and awareness campaigns organized in the départements concerned by the birds’ surveillance (cf § 2). In total, 72 virological analyses were made in the brain of dead birds. All the PCR tests were negative and the West Nile virus has never been isolated. These results suggest that since 2000, no abnormal mortality due to the West Nile virus has been observed in the avifauna of the Mediterranean départements. 3. 2 Serological monitoring of sentinel birds The results of the serological monitoring campaign of “calling ducks” and poultry which were surveyed for four years are presented in the table 2. Table 2. Results of the serosurvey of sentinel birds. Year Nb of controlled birds 288 286 185 % of birds found Nb of sites to be seroposive seroconversions (départements) at the first annual control 28 (dép. 13, 30, 34) 25 (dép. 13, 30, 34) 16 (dép. 13, 30, 34) 9% 2.4% 3.2% 1 duck 1 hen 0 Stes Maries de la Mer (dép. 13) 1 in July 1 in August 6 in September 1 in October St Just (dép. 34) August St Laurent d’Aigouze (dép. 30) October Arles – La Tour du Valat (dép 13) October Place and date of seroconversions Arles (dép. 13) Octobre 2001 Gallargues (dép. 30) August 2002 2001 2002 2003 2004 320 31 (dép. 11, 13, 30, 34, 66, 83) 3.1% (all situated in dép. 83) 9 hens 1 duck 1 duck 2 ducks In the Camargue, between 2001 and 2003, not a single human or equine clinical case has been observed, although a very small circulation of the virus would have been revealed in 2001 and 2002, based on of two cases of seroconversion in birds. Moreover, it is interesting to note that 9% of the birds that have been tested early in the summer of 2001, and 2 to 3% of the birds tested in 2002 and 2003, still showed some serological traces of the 2000 outbreak; this proves a persistance of antibodies that, in birds, may last for several years (the same duck, positive in 2001 was controlled positive in 2003) as well as a lesser degree of seroprevalence after an epizootic peak. In 2003, WN monitoring of sentinel birds of the Camargue was 126 negative. Meanwhile a cluster of WN encephalitis cases was detected in humans (7 cases) and horses (4 cases) in the Var district, located more than 100 km east from the Camargue, in a drier area. Like in the Camargue, no avian mortality was observed in this region. These events showed that, in addition to these known areas, WNV could also appear in other locations in France. In 2004, in the Var region, not a single case of viral circulation was detected anymore, neither in man nor in horses or the avifauna. Nevertheless, the presence of residual antibodies from the 2003 outbreak was still observed in 3.1% of the tested birds that had been checked early in the summer of 2004 in this département. On the contrary, the virus reappeared in the Camargue where the first proof of its presence was noted by the seroconversion of a ‘sentinel’ bird as early as in the month of July 2004 and then by the seroconversion of 12 other birds in the three départements of the Camargue. The circulation of the virus was confirmed by the emergence, in the same area, of 32 clinical cases among horses (10 of which died or have to be euthanized) from September to November. 4 DISCUSSION It seems that the West Nile virus strain that circulates in France since 2000, is not very, or not at all pathogenic for birds because, in the Mediterranean départements which are considered to be areas « at risk », not a single case of mortality due to this virus has been recorded by our surveillance network. The French strain is probably not the same than the one circulating in North America (i.e. this strain is phylogenetically very close to the Israelian one and its pathogenicity for birds has been established) where the presence of dead corvids allow to « be on the virus’ track », something that at the present time is impossible in France. To overcome this difficulty, we have to use sentinel birds that were monitored with serological assays, a method that apparently was efficient and sensitive enough to detect the virus at an early stage and alert the medical and veterinary services. In 2004, the appearance of bird seroconversions before the equine clinical cases, proves that. We can discuss about the lack of sensitivity of the network of sentinel birds that was set up in France and that, with only 320 birds distributed over 31 sites in 6 départements (a limited number because of technical and financial constraints), is, from a numerical and spatial point of vue, theoretically insufficient. Thus, we can suppose that, as soon as it encounters the required environmental conditions, the virus will circulate rather among the mosquitoes and avifauna to be detected in birds by a not very sensitive surveillance network and before the appearance of any clinical signs in horses and humans. From an epidemiological point of view, the results of four years of surveillance make us think that the south of France is an “epizootic area” where limited outbreaks can develop and may be responsible for the contamination of accidental hosts (horses, man), contrary to « enzootic areas » (Africa, Middle East, North America) where the virus largely circulates whereby, each year, new clinical cases appear as well as antibodies that may be found in a great number in its hosts (birds, horses, man….). Even if wetlands, like the Camargue, seem to be more favourable areas for the development of WNV in the reservoirs, vectors and amplifying species, the appearance of the West Nile virus and the intensity of its circulation, that may generate a more 127 or less important risk for human and animal health, are totally unforseeable in our present state of knowledge since they depend on the conjunction of many ecological factors. The phenomenon of global warming could be one of the elements that would explain the emergence of the disease in Europe. Several research programmes have been launched in France and in Europe with the objective to improve our knowledge of the epidemiology of this virus. 5. REFERENCES BALANCA G., HARS J.. Bird reservoirs and indicators of West Nile Fever. Game & Wildlife Science. 2005; in press. 2. CANTILE C., Di GUARDO G., ELENI C., ARISPICI M. Clinical and neuropathological features of West Nile virus equine encephalomyelitis in Italy. Equine Vet. J.2000; 32:31-35 3. CDC. Epidemic/epizootic West-Nile virus in the United States : Guidelines for Surveillance, Prevention and Control. 3rd revision. Centers for Disease Control, 2003; 77 pp. Available online at: http://www.cdc.gov/ncidod/dvbid/westnile/ publications.htm. 4. CHEVALIER V., DURAND B., GERBIER G., BABINOT M., MICHEL JF., TOURE I., ZIENTARA S. 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M., WORK T. H., HURLBUT H. S. & RIZK F. A study of the ecology of West Nile virus in Egypt. Am. J. Trop. Med. Hyg. 1956; 5 : 579-620. 19. TSAI T. F., POPOVICI F., CERNESCU C., CAMPBELL G. L., NEDELCU N. I. West Nile encephalitis in southeastern Romania. The Lancet 1998 ; 352:767-71. 20. WEINBERGER M., PITLIK S. D., GANDACU D., LANG R., NASSAR F., BENDAVID D., RUBINSTEIN E., IZTHAKI A., MISHAL J., KITZES R., SIEGMANIGRA Y., GILADI M., PICK N., MENDELSON E, BIN H, SHOHAT T, CHOWERS MY, West Nile fever outbreak, Israel, 2000: epidemiologic aspects. Emerg. Inf. Dis. 2001; 7:679-685. 21. WORK T.H., HURLBUT H.S., TAYLOR R.M. Isolation of West Nile virus from hooded crow and rock pigeon in the Nile Delta. Proc Soc Experiment Biol Med .1953; 84: 719-72. 22. ZELLER H.G., SCHUFFENECKER I. West Nile Virus: An Overview of Its Spread in Europe and the Mediterranean Basin in Contrast to Its Spread in the Americas. Eur.J. Clin. Microbiol. Information. Dis. 2004 (in press). 23. ZELLER H. & MURGUE B. Rôle des oiseaux migrateurs dans l’épidémiologie du virus West Nile. Med. Mal Infect. 2001 ; 31 : 168-174. 24. ZIENTARA S., MURGUE B., ZELLER H., DUFOUR B., MURRI S., LABIE J., DURAND B. & HARS J. Maladie à virus «West Nile» en France. Epidémiologie et Santé Animale 2001 ; 11 : 295-298. AUTHORS ADRESSES Jean Hars Unité sanitaire de la faune, Office national de la chasse et de la faune sauvage, 5 Allée de Bethléem, 38610 Gieres, France Email:
[email protected] 129 Great Western Referrals1, Avian and Exotic Department, Swindon, United Kingdom; Veterinary Medical Research Institute2, Hungarian Academy of Science, Budapest; Central Veterinary Institute3 Budapest, Hungary; Institute for Virology4, Faculty of Veterinary Medicine University of Leipzig, Germany ADENOVIRUS INFECTION IN RAPTORS P. Zsivanovits1 DMedVet MRCVS, D. J. Monks1 BVSc (Hons) MACVSc (Avian Health) CertZooMed MRCVS, N. A. Forbes1 BVetMed CBiol MIBiol Dip ECAMS FRCVS, M. Benk� DMedVet PhD2, K. Ursu MRCVS2,3, R. Raue4 DMedVet KEYWORDS Adenovirus - Ventricular ulceration - Intranuclear inclusion bodies - Raptors ABSTRACT Although previously described as a cause of mortality, adenovirus remains an under-reported disease in raptors. This report describes adenoviral outbreaks in two separate collections at similar times, involving a Harris hawk (Parabuteo unicinctus), a Bengal eagle owl (Bubo bengalensis), and a Verreaux’s eagle owl (Bubo lacteus). The outbreaks were diagnosed by necropsy, histologic examination, and PCR. Attempts to isolate virus or to detect viral particles by electron microscopy were unsuccessful. PCR with consensus primers resulted in amplicons of specific sizes. DNA sequencing identified the detected virus as a member of the genus siadenovirus. To the author’s knowledge this is the first report of disease related to adenovirus infection in these species. This report highlights the clinical and histologic presentation of adenoviral infections, and the difficulty of diagnosis. 1 INTRODUCTION Adenoviruses are non-enveloped double-stranded DNA viruses, 70-90 nm in diameter, that replicate in the cell nucleus, forming basophilic intranuclear inclusion bodies. Based on genetic and phylogenetic analysis, four genera of adenoviruses are recognised (BENK� et al. 2004). Mastadenoviruses affect mammals. Aviadenovirus (former group I) are isolated from numerous bird species including poultry. One novel genus, Siadenovirus, consists of the ‘Turkey haemorrhagic enteritis virus’ (‘Turkey adenovirus type 3’), ‘Marble spleen disease virus’ and ‘Chicken splenomegaly virus’, originating from different diseases in different species but indistinguishable on 130 serology, together with an adenovirus isolated from a frog. ‘Egg drop syndrome’76 – virus’ (‘Duck adenovirus type 1’) together with multiple adenoviruses isolated from reptiles, birds, ruminants and a marsupial form the other new genus, Atadenovirus (BENK� et al. 2004). Adenoviruses are reported to be opportunistic, depending on triggering factors (such as immunosuppression by bacterial, fungal, viral or parasitic pathogens) to cause clinical disease. (RITCHIE 1995). There have been sporadic reports of adenovirus infections in raptors with varying clinical signs: a Northern goshawk (Accipiter gentiles) was found with central nervous signs (STEHLE 1965, cited by GERLACH 1994); American kestrels (Falco sparverius) (SILEO et al. 1983) and a tawny frogmouth (Podargus strigoides) (REECE and PASS 1985) were diagnosed with haemorrhagic enteritis and splenomegaly; a merlin (Falco columbarius) showed adenoviral hepatitis (SCHELLING et al. 1989); and fatal adenovirus infections were reported in Mauritius kestrels (Falco punctatus) (FORBES et al. 1997). This paper discusses adenoviral outbreaks in two raptor collections. In one collection Harris’ hawks (Parabuteo unicinctus) were primarily affected, while in the second collection a Bengal eagle owl (Bubo bengalensis) and a Verreaux’s eagle owl (Bubo lacteus) died. Identification of the causative agent was attempted based on the results of clinical examination, necropsy, histologic examination, virus isolation, electron microscopy and PCR. This report highlights the difficulties in diagnosing adenoviral infection, particularly in the acute stages. To the best of the authors’ knowledge this is the first report of adenovirus infection in a Harris hawk, a Bengal eagle owl or a Verreaux’s eagle owl. 2 CASE REPORT The death of the 20-weeks old Harris hawk was preceded by approximately ten minutes of fitting. The 3-year-old European eagle owl showed 24 hours of increasing depression and the 1-year-old Bengal eagle owl showed 48 hours of depression and anorexia prior to death. Necropsy findings of the Harris hawk and the eagle owls included hepatomegaly; splenomegaly; proventricular and ventricular dilation, ulceration and erythema; and renomegaly. The Harris Hawk had a Syngamus spp. infestation, while the Verreaux’s eagle owl had a suspected protozoan infection. Histological findings of all three birds consisted of hepatic necrosis, hepatitis, splenic necrosis, proventricular and ventricular ulceration and necrosis. Besides the above-mentioned organs, basophilic intranuclear inclusion bodies were also seen in the pancreas and the kidneys of the Verreaux’s eagle owl. Virus isolation on chicken embryo liver cells and electron microscopy of pooled tissue samples (liver, spleen, ventriculus, kidney, heart) of the Harris hawk and the Eagle owls were negative for adenovirus. PCR was performed on pooled tissue samples in two laboratories, one in Leipzig, Germany, and one in Budapest, Hungary. Both laboratories carried out Aviadenovirus-specific PCR to detect the 12 fowl adenovirus 131 serotypes (former group I) using hexon gene-targeting primers (RAUE and HESS 1998), even in a nested system. None of these detected adenoviral DNA in the samples. The Budapest laboratory also performed nested PCR with a primer targeting a partial sequence of the polymerase gene (WELLEHAN et al. 2004). This PCR detected adenoviral DNA in tissue samples of all three carcasses submitted. DNA sequencing of the PCR products revealed that all three birds were infected with the same virus and alignment of amino acid sequences classified it as a member of the novel genus, siadenovirus. On the phylogenetic tree (based on the results of distance matrix analysis), the virus was grouped with Siadenoviruses, in a common branch with the ‘Turkey adenovirus 3’, and with the frog Siadenovirus representing a separate branch. Consensus PCR for circovirus and polyomavirus and a nested PCR for herpesvirus were also performed on the pooled tissue samples in the Leipzig laboratory and no viral DNA was detected. 3 DISCUSSION The large basophilic intranuclear inclusion bodies of varying size coupled with the distribution in epithelial and lymphoreticular tissue, karyomegaly, hepatic and splenic necrosis and ventriculitis combine to generate a histological pattern that is highly suggestive of adenoviral infection in the presented cases. Intranuclear inclusion bodies were identified in the liver, spleen, ventriculus, pancreas, small intestines and kidneys. This pattern has also been observed in poultry infected with adenovirus (NAKAMURA et al. 2002; RITCHIE 1995). The Harris hawk belonged to a clutch of four fledglings that were kept free-lofted in a large aviary with their parents. Prior to the death of this Harris hawk the other three fledglings died acutely with no premonitory signs within a period of 21 days. During the period of Harris hawk mortality, there were also two 1-year-old red kites (Milvus milvus) dying in the same collection. Those deaths were also acute with no premonitory signs. On necropsy those two red kites as well as the first three dead Harris hawks were in good body condition and showed petechial haemorrhages and inflammation of the ventricular wall and the myocardium, pancreatic congestion and haemorrhages, and moderate splenomegaly. Histologic findings of the kites and the first three dead Harris hawks were non-specific with severe acute multifocal to coalescent myocardial necrosis and haemorrhage, and moderate to severe acute lymphoid depletion in the bursa and spleen. Inclusion bodies were not detected. Those histologic findings were consistent with any acute bacterial or viral infection. However, based on the similar presentation on necropsy and the close timing of the deaths, adenoviral infection as cause of death in the initially affected Harris hawks and the red kites is highly suspicious. The clinical course of this outbreak with non-specific initial necropsy findings such as haemorrhages and inflammation of internal organs, and intranuclear inclusion bodies becoming more apparent during the course of infection is consistent with a previous report (FORBES et al. 1997). Experimental infections of broilers with adenovirus showed that the frequency of intranuclear inclusions was greatest after five days post-inoculation and that histologic lesions such as ventricular erosions appeared seven to nine days after inoculation (NAKAMURA et al. 2002). 132 This progression reiterates how difficult it can be to make an ‘adenovirus’ diagnosis when only a single bird is affected and dies following a peracute or acute infection. It demonstrates the potential need of serial necropsy and histologic examinations of subsequent losses within an outbreak to elucidate the aetiology of an infection within a collection. It is notoriously difficult to isolate adenovirus infecting raptors on chicken embryo liver cells (GOUGH; GERLACH, personal communication, 2004). Atadenoviruses and Siadenoviruses show generally poor in vitro replication ability. The frog Siadenovirus could only be propagated on turtle heart cells, while ‘Turkey adenovirus type 3’ can be propagated in embryos or young birds, but not in conventional cell lines (BENK� et al. 2004). Electron microscopy requires a virus particle concentration of greater than one million virus particles per millilitre of sample to give positive results (RITCHIE 1994). Therefore, negative results of virus isolation and electron microscopy do not exclude adenoviral infection. PCR is a rapid and sensitive way to screen clinical samples for the presence of microbial DNA. It also facilitates the acquisition of DNA templates for sequencing. Hexon is the major component of the adenovirus capsid. Although providing several highly conserved regions, there is considerate variation in members of the different genera. For conventional Aviadenoviruses, several hexonspecific PCR primers have been described (RAUE and HESS 1998), which gave negative results in the present cases. Consensus, highly degenerated primers for the detection of a short fragment of the adenoviral DNA-polymerase gene as well as an even more sensitive, nested PCR system, targeting the neighbouring DNA-polymerase gene, have been described (WELLEHAN et al. 2004). In the Budapest laboratory, this nested PCR system was used to generate DNA fragments for sequencing. The taxonomic place of this likely new adenovirus type is as yet preliminary, and needs further confirmation with the use of Siadenovirus-specific primers. As inclusion bodies of circovirus, polyomavirus and herpesvirus can resemble adenoviral inclusion bodies (RITCHIE 1995), further PCR was performed on the tissue samples to exclude those viruses. It is interesting that adenovirus was identified in Harris hawks and eagle owls, representing species that are often considered as less susceptible to a number of bacterial or viral diseases, compared with less ‘hardy’ species such as American kestrels or Mauritius kestrels. One report states a high susceptibility of Mauritius kestrels to infection with adenovirus group I (Aviadenovirus) (FORBES et al. 1997). However, during the outbreaks there were no deaths in any kestrel species that were also kept in the collection. It is known that different serotypes of adenovirus express different pathogenicity and can pose a threat to different host-species (RITCHIE 1995, FORBES et al. 1997). Certain adenoviruses, especially the members of the genera atadenovirus and siadenovirus, have proved to be highly pathogenic, being capable of infecting multiple host species and causing experimentally reproducible specific diseases (BENK� et al. 2004). The supposedly higher pathogenicity of Siadenovirus involved in these cases compared to Aviadenovirus serotypes might explain the development of disease in these new host species. It is also possible that the endoparasite infections found in some of the birds may have immunocompromised them such that the adenovirus infection could become clinically apparent. 133 Free-ranging pigeons and waterfowl may present a reservoir for adenovirus (RITCHIE 1995). However, the two collections are 78 km apart from each other and to our knowledge there were no reports about other deaths in birds due to adenovirus infection in proximity of those collections. Day-old chicks infected with adenovirus were found responsible for fatal adenovirus outbreaks in Mauritius kestrels (FORBES et al. 1997). The two collections described in this report were using the same food source to purchase their day-old-chicks. No further birds died in either collection after both were advised to feed mammalian-derived food items to their birds for the next few weeks. Prevention of adenovirus infection is difficult. Simply avoiding feeding avian-derived food is relatively reliable, but this is difficult for larger collections with respect to practicability and costs. However, when confronted with unexplained deaths within a collection of raptors, the immediate halt of feeding all avian-derived food is recommended. As wild pigeons or waterfowl can also serve as source of adenovirus infection, strict measurements of hygiene including the prohibition of any contact with wild birds or their faeces are necessary to help prevent disease outbreak in collection of birds of prey. To control adenoviral disease molecular biological techniques such as PCR and serology on avian-derived food items can be considered. However, the extensive variety of different genera and serotypes is likely to complicate matters. If a sufficiently common adenoviral antigen could be found, then vaccination of raptors might be an interesting strategy. CITATION INDEX 1. 2. 3. 4. 5. 6. 7. 8. 9. BENK� M, HARRACH B, BOTH GW, et al (eds): Virus taxonomy, VIIIth report of the International Committee on Taxonomy of Viruses., London: Elsevier/Academic Press 2004; 213 - 228. FORBES NA, SIMPSON GN, HIGGINS RJ and GOUGH RE. Adenovirus infection in Mauritius kestrels (Falco punctatus). J Avian Med Surg 1997; 11(1): 31 - 33. NAKAMURA K, OHYAMA T, YAMADA M, et al. Experimental gizzard erosions in specific-pathogen-free chicks by serotype 1 group I avian adenoviruses from broilers. Avian Dis 2002; 46(4): 893 - 900. RAUE R and HESS M. Hexon based PCRs combined with restriction enzyme analysis for rapid detection and differentiation of fowl adenoviruses and egg drop syndrome virus. J Virological Methods 1998; 249(2): 37 - 315. REECE RL and PASS DA. Inclusion body hepatitis in a tawny frogmouth (Podargus strigoides). Aust Vet J 1985; 62: 426. RITCHIE BW. Adenoviridae. In: RITCHIE BW (ed): Avian viruses – Function and control. Lake Worth: Wingers Publishing 1995; 313 - 334. SCHELLING SH, GARLICK DS and ALROY J. Adenoviral hepatitis in a Merlin (Falco columbarius). Vet Pathol 1989; 26: 529 - 530. SILEO L, FRANSON JC and GRAHAM DL. Hemorrhagic enteritis in captive American kestrels (Falco sparverius). J Wildl Dis 1983; 19: 244 - 247. STEHLE S. Krankheiten bei Greifvögeln (Accipitres) und bei Eulen (Sträges) 134 Tiermedizinische Dissertation. 10. HARRACH B. Tierärztliche Hochschule Hannover. et al. 78(23): 13366 . 862 .uk 135 . HARRISON GJ and HARRISON LR (eds): Avian Medicine: Principles and Application. Great Britain Email: P. Berkshire House. Detection and analysis of six lizard adenoviruses by consensus primer PCR provides further evidence of a reptilian origin for the Atadenoviruses. JOHNSON AJ. AUTHORS ADDRESS P. Germany. Shrivenham Road. J Virol 2004. SN1 2NR.co. Avian and Exotic Department. In: RITCHIE BW.Zsivanovits@gwreferrals. Viruses. WELLEHAN JFX.13369. County Park Estate. Lake Worth: Wingers Publishing 1994. Unit 10. Swindon.mit Ausnahme der parasitären Erkrankungen. Zsivanovits DMedVet MRCVS Great Western Referrals. Cited by GERLACH H.948. 1965. K.Birds . even when vitamin C or a meat-based dog food was added.Treatment . Department of Pathobiology2. PhD KEYWORDS Iron storage disease . The addition of inositol and tannic acid to the high iron diet prevented an increase in liver iron. or low iron with vitamin C. College Station. when stained for iron.Phlebotomy . DVM.Diet . Phalen1. Russell2. percent iron saturation and staining for iron in bone marrow aspirates were not found to be effective predictors of liver iron stores. The low iron diet alone reduced nonheme liver iron levels at a much slower rate. However. The high iron diet caused increased liver iron concentrations to concentrations similar to those found birds that have died of iron storage disease (ISD). PREVENTION AND TREATMENT OF IRON STORAGE DISEASE USING THE EUROPEAN STARLING AS A MODEL D.Deferoxamine . low iron with a meat-based dog food. hepatocytes obtained from liver core biopsies. An iron concentration of 32 ppm in the diet was found to be inadequate to meet the physiological demands of the starlings.Diagnosis . United States of America DIAGNOSIS. or the addition of inositol and tannic acid to the low iron diet. phlebotomy. iron binding capacity. G. were found to accurately predict liver iron concentrations. N. Birds were then switched to low iron diets and treated with the low iron diet only. These results suggest that both phlebotomy and treatment with deferoxamine are effective treatment options for birds with ISD. Texas A&M University. OLSEN1. BA. The addition of inositol and tannic acid to the low iron diet had no impact on nonheme liver iron concentrations. Department of Pathobiology. or low iron diets and either deferoxamine.Schubot Exotic Bird Health Center1. In a second experiment.Liver iron ABSTRACT Iron absorption was compared in European starlings fed diets containing high iron. starlings were fed an iron loading diet to induce high liver iron concentrations. Total plasma iron.Prevention . Phlebotomy and deferoxamine treatments reduced nonheme liver iron concentrations to safe levels in 16 weeks of treatment at similar rates. Texas A&M University. high iron with inositol and tannic acid. low iron. 136 . PhD. DVM. and bone marrow aspirates for predicting liver iron concentrations. METE et al. and they are fed diets containing excess iron in captivity (WARD et al. 2001). DORRESTEIN et al. mynahs (Sturnidae). Left untreated ISD is fatal. but appears to be less than 135 ppm. Once high liver iron concentrations were obtained. 2004a). DORRESTEIN et al. Starlings on the high iron diet and high iron diet supplemented with inositol and tannic acid were euthanased at 6 and 16 weeks. It is also possible that vitamin C and meat may enhance iron absorption (ROSSANDER-HULTH and HALLBERG 1996). CRISSEY et al. Finally. 2003. low iron diets containing a phytate and tannin. 2004). The actual safe and adequate concentration of iron in these birds’ diets is not known. 1991. Control starlings were euthanased immediately. 2 MATERIALS AND METHODS Effect of supplements on liver iron concentrations Iron absorption was compared in European starlings (Sturnus vulgaris) fed diets containing high iron (1585 ppm). ISD appears to occur in these species of birds because they are very efficient at absorbing iron from their diets. 1991. 2000). and low iron diets in combination with deferoxamine administration or phlebotomy. 2000. Effect of treatment on liver iron concentrations European starlings were fed an iron loading diet (3235 ppm) for 31 days to induce nonheme liver iron concentrations approaching those in birds with that died with iron storage disease. 137 . The pathogenesis of ISD is poorly understood but ultimately appears to result in heart failure and may in some cases cause liver disease. CRISSEY et al.1 INTRODUCTION Iron storage disease (ISD) is a life threatening disease of captivity that occurs in several species of toucan (Rhamphastidae). we examined the usefulness of plasma chemistry values. or low iron with vitamin C. high iron with inositol (a phytate) and tannic acid. liver aspirate. low iron with a meat-based dog food. The second phase of the experiment was to look at the effectiveness of treating birds with with high liver iron concentrations with low iron diets. The first part of this study was designed to determine what the minimum dietary concentration of iron is and what effect supplementation would have on iron absorption. GAFFNEY et al. 2000. 2000. low iron (32-34 ppm). they do not down regulate iron absorption when iron replete. all birds were switched to low iron diets (32 to 48 ppm) and divided into four treatment groups. Iron storage disease is characterized by a massive accumulation of iron in the liver (ROSSANDER-HULTH and HALLBERG 1996).) (WARD et al. It is possible that addition of phytates and/or tannins to these bird’s diets would reduce iron absorption (SEIBELS et al. birds of paradise (Paradisaeidae) and quetzals (Pharomachrus spp. The remaining birds were euthanased at 16 weeks (OLSEN et al. 138 . even when vitamin C or a meat-based dog food was added and liver iron concentrations decreased in all birds on these three diets. Novartis Pharmaceuticals. The addition of inositol and tannic acid to the low iron diet had no impact on nonheme liver iron concentrations (OLSEN et al. Bone marrow and liver samples were stained with Prussian blue for iron and the number of iron containing cells and the intensity of their iron staining was recorded (RUSSELL et al. Desferal. Prediction of liver iron concentrations in the live bird Prior to euthanasia. Starlings were treated for 16 weeks (OLSEN et al. The low iron diet alone reduced nonheme liver iron levels at a slower rate (45ppm/week). Total plasma iron. The addition of inositol and tannic acid to the high iron diet prevented an increase in nonheme liver iron (OLSEN et al. 2004a). 3 RESULTS Effect of supplements on liver iron concentrations The diet with low iron concentration (32 ppm) was found to be inadequate to meet the physiological demands of the starlings. SQ. Nonheme liver iron concentrations for both the supplement and treatment experiments were determined using a modification of the Torrance-Bothwell nonheme method and 0.5 g of liver obtained at necropsy from each starling (OLSEN et al. East Hanover.05). 2004b). and liver core biopsies were obtained by repeated transabdominal insertions of a 23 gauge needle. birds were anesthetized and blood was collected from the right jugular vein. phlebotomy (1% of body q 7 d). The high iron diet caused increased nonheme liver iron concentrations to concentrations similar to those found birds that have died of ISD. NJ. 2004a). q 24 hr.One treatment group was treated with the low iron diet only. or the addition of a phytate (inositol) and tannic acid to the low iron diet. bone marrow was aspirated from the tibiotarsus. Effect of treatment on liver iron concentrations Phlebotomy and deferoxamine treatments reduced nonheme liver iron concentrations to safe levels in 16 weeks of treatment at similar rates (190 ppm/week and 163 ppm/ week respectively). 2005). 2004b). iron binding capacity. and percent saturation was determined. Statistical analysis Mean liver nonheme iron concentrations and mean weights for controls and each treatment group were compared using the two tailed t-test (P<0. Core biopsies of the liver were expressed onto glass slides by forcing air through the needle. USA). Other treatments groups were treated with either deferoxamine (100mg/kg. The treatment experiment suggests that both phlebotomy and treatment with deferoxamine are effective treatment options for birds with ISD. DORRESTEIN G. Based on necropsy findings in birds that had this procedure.233-238. birds with moderate liver iron concentrations. 2. They also suggest that the addition of a phytate or tannic acid or both to commercial diets may be an inexpensive way to prevent ISD in captive birds. Portland 2000. J Zoo Wildl Med 2000. Proc Conf Assoc Avian Vet. these additives did not accelerate a decline in liver iron concentrations. METE A. It is possible that a combination of these treatments or an increased frequency of phlebotomy would result in a more rapid decline in liver iron concentrations and still be safe for the bird. Fine needle core biopsies of the liver were found to be the only effective means of approximating the liver iron concentrations in the live bird. BLOCK S and MASLANKA M. Although a combination of phytates and tannins prevented excess iron absorption. 5 ACKNOWLEDGEMENTS This work was funded by a generous donation from the Morris Animal Foundation and the Schubot Exotic Bird Health Center. 6 CITATION INDEX 1. Iron stained preparations of the biopsy material consistently allowed the differentiation between birds will low to normal liver iron concentrations. LEMMENS I and BEYNEN A. this technique appeared to be safe. 139 . Hemochromatosis/ iron storage: new developments. 31: 491-496. Hepatic iron accumulation over time in European starlings (Sturnus vulgaris) fed two levels of iron. The low iron diet was considered a useful adjunct to the treatment of ISD by other methods. and birds with high iron concentrations (RUSSELL et al. 2005). Bone marrow aspirates contained very few iron positive cells and also could not be used to predict liver iron concentrations. These results also suggest that supplementation of diets that contain adequate iron with small amounts of fruit or meat will not significantly impact iron absorption. WARD A.Prediction of liver iron concentrations in the live bird Plasma iron. CRISSEY S. but was considered to be an inadequate by itself. Core biopsy of the starlings’ liver was found to be safe and in the majority of attempts resulted in a sample adequate for analysis. 4 DISCUSSION The results of the additive experiments suggest that the minimum and safe dietary iron concentration for birds susceptible to ISD is close to 50 ppm. iron binding capacity and percent saturation where of no value in predicting the nonheme liver iron concentrations. WARD R. SEIBELS B. Phalen. 3. and PHALEN DN. et al.115. DIERENFLED E. Bios 2004. Dietary factors influencing iron absorption . 7.53.an overview. A comparison of four treatment regimens for iron storage disease using the European starling (Sturnus vulgaris) as a Model. DIERENFELD E. AUTHORS ADDRESS David N. ABVP (Avian) Schubot Exotic Bird Health Center.316. 75: 43 .GAFFNEY S. MARX J. 20: 225 . 8. Avian Pathol 1991. 10. RUSSELL K. 2005. RUSSELL K. HENDERSON G and PETERS T. et al. 105 . OLSEN GP. London: John Libbey & Company 1996. Submitted: J Avian Med Surg 2004. WILLIAMS V. PhD. OLSEN GP. 30: 479 . ROSSANDER-HULTH L and HALLBERG L. Submitted: J Avian Med Surg 2004. Manuscript in preparation. LAMBERSKI N. DVM. et al. A comparative study of iron retention in mynahs.tamu. et al. doves. METE A. DORRESTEIN G. RUSSELL K. Texas A&M University. FLYNN P. Avian Pathol 2001. TX 77843-4467. 6. DIERENFELD E and PHALEN DN. Effective use of tea to limit dietary iron available to starlings (Sturnus vulgaris). College Station.486. In: Iron Nutrition in Health and Disease. Tannin/polyphenol effects on iron solubilization in vitro. J Zoo and Wildl Med 2003. United States of America Email: dphalen@cvm. SMITH T.232. OLSEN GP. Impact of supplements on iron absorption from diets containing high and low iron concentrations in the European starling (Sturnus vulgaris). Investigation of the aetiology of haemosiderosis in the starling (Sturnus vulgaris). 5. GREGORY C. 4. 9. Dipl.edu 140 . 34: 314 . and rats. toucans and birds of paradise and is rarely seen in these same species in the wild.Department of Animal Science1. Pesaro1. especially in liver. Private Practitioners2. In fact this disease has been most commonly seen in captive soft billed birds such as mynahs. The study demonstrates the feasibility of an experimental animal model for the evaluation of the progression to the hepato-cellular carcinoma in the human genetic disease by using the mynah birds. Petroselli2 KEYWORDS Haemochromatosis . close to HLA gene complex (BACON and HARRISON 2003). This pathology is rarely shown in animals. Genetic haemochromatosis is an autosomal recessive disorder in which the gene of the disease is located on the short arm of chromosome 6. Peccati2. The present study tries to demonstrate the different steps in the disease progression. Because humans do not have a major excretory pathway for iron. cellular proliferation and neo-plastic transformation of the hepatocytes involved in different degrees of iron overload. S. Rossi1. ABSTRACT Haemochromatosis is a disease characterised by an excessive enteric absorption of iron. 2001).Cirrhosis .Tumour markers. Italy BIO-MOLECULAR STUDY OF A PROGRESSION OF HAEMOCROMATOSIS IN HILL MYNAHS (GRACULA RELIGIOSA) G. by showing the expression of different and specific marchers of fibrosis. In genetic 141 . subsequently stored in different organs.Hill mynahs (Gracula Religiosa) . Studies show that hepatic iron overload may be one of the most important causes that induce the progression to liver fibrosis and hepatocarcinoma. University of Camerino. P. It seems to occur more commonly in frugivores and insectivores. pancreas. haemochromatosis results either from a genetic defect causing excessive iron absorption or as consequence of parental administration of iron (BACON et al. There is growing evidence to indicate that lories may also be a species vulnerable to this disorder.Hepatocarcinoma . kidney and heart (BACON and HARRISON 2003). with the exception of some species of birds. DNA alteration. 1 INTRODUCTION In human beings haemochromatosis is characterised by excessive accumulation of body iron. most of which is deposited in the parenchymal organs such as liver. R. Ceccherelli2. C. anti AFP (monoclonal antibody. The aim of the study is the evaluation of the levels of expression of different and specific marchers of fibrosis. For morphological and histochemical evaluations.. University of Camerino – Italy). 2 MATERIALS AND METHODS During a period of three years. Fisher. and Van Gieson trichrome stain for evaluation of various degrees of fibrosis. Samples of liver. Novocastra. anti PCNA (monoclonal. There is growing evidence to indicate that lories may also be a species vulnerable to this disorder. were routinely deparaffinised and then stained with hematoxylin and eosin. followed by a proteinase K (DAKO Corporation. In four animals. London ). Novocastra. London). histological sections. placed in 10% buffered formalin. 142 . Two mynahs and a blackbird that died from trauma without any signs of hepatic disease and normal biochemical parameters were used as control birds. PA). Burlingame. leading to net iron accumulation. leading to lethal injury or predisposition to hepato-cellular carcinoma (ELMBERG et al. anti CEA (monoclonal antibody. lung. Prussian’s blue stain (specyfic to dye Fe+++ into the cells). anti CA19. cellular proliferation and neoplastic transformation in the hepatocytes. In our experience. dyspnoea and cardiac dysfunction (arrhythmia and cardiomyopathy). proventriculus. 35 hill mynahs (Gracula religiosa) and 1 blackbird (Turdus merula) were necropsied in our Laboratory (Department of Veterinary Sciences. a moderate cardiomegaly was also seen. Novocastra. kidney. the same deparaffinised sections were immersed in distilled water containing 15% H2O2 for 30 min at room temperature in order to inactivate endogenous peroxidases. heart. Pittsburgh. In mynahs the clinical features of the disease are very similar to those observed in human genetic or primary forms. stimulation of collagen formation. 2003). The sections were then stained with primary anti mutate-P53 (monoclonal antibody. In order to minimize nonspecific background staining. CA) treatment at room temperature for 6 min. ventriculus. some birds with severe cirrhosis or hepatocelluar carcinoma showed high degree of iron stored into hepatocytes. Santa Cruz. mounted on positively charged glass slides (Superfrost Plus. pancreas. DNA alteration. 1994) and is rarely seen in these same species in the wild. ascites at necropsy. sometimes. UK). and paraffin embedded. spleen. in terminal phases. STYLES 1999). The other mynahs examined. Excessive iron seems to be directly toxic to tissues by different mechanisms as: lipid peroxidation by iron-catalyzed free radicals reactions (BACON and BRITTON 1990). Dako Co. For immunohistochemical tests. Carpinteria. Haemochromatosis in animal is infrequent conditions with the exception for some species of birds (DIERENFELD et al. nodularity of the parenchyma and.haemochromatosis there seems to be a primary defect in the intestinal absorption of dietary iron. the sections were incubated in a serum blocking solution (10% non-immune horse serum in 1% bovine serum albumin dissolved in PBS) for 1 hr at 37 °C. showed alterations in biochemical values and different degrees of hepatomegaly. In fact this disease has been most commonly seen in captive soft billed birds such as mynahs. bursa and bone-marrow were collected from all birds. It seems to occur more commonly in frugivores and insectivores. ascites and. The slides were rinsed in phosphate-buffered saline (PBS) three times for 2 min each. liver fibrosis. abdominal dilation. California). small intestine. and anti Collagen type IV (polyclonal rabbit anti chicken. related to different degrees of iron stored into hepatocytes and other cellular types. toucans and birds of paradise (DIERENFELD et al. London).9 (monoclonal antibody. hypoalbuminaemia. In chronic cases the principal manifestations include hepatomegaly. 1994. and direct interactions of iron with DNA. 143 . Results of necropsies are summarised in table 1. P53. score 3= diffuse PB stain of hepatocytes with strong stain in groups of perivascular macrophages and Kupffer cells – presence of extrahepatic hemochromatosis (PB stain of myocardiocytes. and the mean of cells positive for PCNA. ten 40x randomly selected fields were analysed for each sample and the number of positive cells/field was recorded. Negative controls included apparently normal mynahs in which Prussian’s Blue (PB) reaction gave negative results in liver’s sections and tissues of black bird that represent a bird’s species not involved in pathological iron overload. followed by incubation with 3. the degree of Fe+++ stored in the hepatocytes. 3 RESULTS Macroscopic examination of necropsied birds revealed various degrees of morphological alterations of the liver in 30 of 36 (83. renal epithelium and septal macrophages of the lungs).5%) Hepatocarcinoma 4 (11. accompanied by nodules of hepatocytes regeneration. sections were incubated with enzyme conjugate (ABC-peroxydase complex. rapidly counterstained with haematoxylin and examined microscopically.Santa Cruz. score 3= cirrhosis with severe degree of perivascular and intercellular fibrosis. varying degrees of ascites were observed and only in three cases a moderate cardiac dilation was also present. CEA and CA19.5%) 9 (25%) 4 (11. After three rinses in PBS.1%) Table 2 reports the data regarding the morphological aspect of the livers. rinsed in PBS and incubated with biotinylated secondary antibody (biotinylated anti-mouse or anti-rabbit immunoglobulins) for 45 min at room temperature. Burlingame. UK) for 45 min at room temperature. score 1= mild degree of perivascular fibrosis. Optical quantification of different degrees of intracellular iron stored were performed by scoring the Prussian’s Blue dye using the following scores: score 0= absence of iron in hepatocytes. the degree of fibrosis express as degree of VG and collagen IV stains. Similar scale was used for quantification of different degrees of fibrosis in Van Gieson (VG) and Collagen type IV stained slides: score 0= absence of fibrosis. score 2= moderate degree of perivascular fibrosis with irradiation of collagen between hepatocytes.9 per sample. The slides were rinsed in distilled water. Vector.6%) 11 (30. CA).3’-diaminobenzidine solution containing H2O2 as substrate-chromagen for 5 min. score 1= groups of hepatocytes with weak PB stain. All antibodies were diluted 1:50 in PBS and incubated overnight in a 4 °C moist chamber. score 2= diffuse PB stain of hepatocytes. The value of expression of each marker per sample was expressed as the mean value obtained from these ten values. All values obtained were statistically analysed and compared by using the T Student test. For immunohistochemical evaluation. In almost the 36% of analysed animals. In some cases the most evident alterations were represented by dark colouring and enlargement of the liver.1%) 2 (5.4%) examined animals. Table 1: Macroscopic appearance of the liver in necropsied birds Examined Normal Hepatomegaly Liver Cirrhosis Cirrhosis with birds liver fibrosis neoplastic lesions 36 (100%) 6 (16. AFP. Fat degeneration Congested with fatty spot liver. Different areas of fat degeneration. Moderate degree of congestion Van Perl Gieson score score 0 0 0 1 0 1 1 0 1 1 0 0 0 0 0 0 0 0 0 0 PCNA P53 3 7 2 1 4 0 3 5 8 23 0 2 0 1 0 0 1 0 2 5 A F P 1 2 0 0 1 0 4 0 5 7 C E A 0 1 0 0 2 0 1 0 2 32 CA Collagen 19. Chronic hepatitis Perivascular fibrosis with congested liver.Table 2: Morphology and score of different parameters evaluated in bird’s liver. Chronic-active hepatitis with perivascular infiltrate Chronic-active hepatitis with perivascular infiltrate Normal Chronic hepatitis with diffuse perivascular accumulation of macrophages. spotted in color 6439/B Hepatomegaly. discoloured B1504/A hepatomegaly.9 IV 0 0 0 0 0 0 1 0 0 0 0 0 0 0 0 0 0 0 1 0 2 1 30 8 11 27 2 1 1 2 3 1 1 1 0 1 3* 2 1 0 1 0 2 1 0 0 12 45 51 12 5 3 0 7 12 3 0 53 39 7 0 2 0 1 3 15 9 11 0 4 0 0 10 18 33 42 17 7 0 0 5 6 0 5 8 0 0 0 0 0 2 2 1 1 1 0 3 1 0 1 144 . Acute hepatitis with severe heterophilic infiltrate. Case 2687/3 Control Turdus merula 2681/a control normal mynah 2190/A control normal mynah 2233/c apparently normal mynah 632/1 apparently normal mynah 528/B apparently normal mynah 369 hepatomegaly 571 hepatomegaly 2494B hepatomegaly 2231 hepatomegaly 188/v hepatomegaly 670 hepatomegaly B2494/1 hepatomegaly B2494/2 hepatomegaly dark color B2494/3 hepatomegaly 805/B hepatomegaly congested B2156 hepatomegaly. congested B2982/A hepatomegaly Hematoxylin and eosin Normal Normal Normal Normal Normal Normal Normal Normal Groups of hypertrophic hepatocytes. 669A(B) Carcinoma Moderate degree of fibrosis Mild fibrosis Moderate degree of fibrosis associated to severe hepatocellular atrophy Mild fibrosis. 342/F Atrophy of right lobe 589/3 Normal B/2345 Atrophy 2122 Normal 1532/M Mild atrophy 783 Mild atrophy 3338 Atrophy of the left lobe B1928 Cirrhosis 2156/1 Cirrhosis B 3345 Cirrhosis B 2521 Cirrhosis B2156 Cirrhosis with umbilicate nodules B2222A Carcinomatous nodules 669A(A) Carcinoma.B2222B Morphological alteration. Mild fibrosis Regenerative nodules and fibrosis with severe morphological alterations Morphological alterations Regenerative nodules Marked morphological alterations Macronodular cirrhosis with hepatocarcinoma and chronic active hepatitis. with fibrosis *Extrahepatic haemochromatosis characterized by myocarditis. 1 2 2 1 6 22 0 5 13 7 11 9 0 0 3 2 3 2 9 11 5 12 1 2 2 3 1 2 3 2 1 3 1 1 2 1 13 10 5 11 21 4 1 13 0 2 11 7 3 18 0 12 18 23 0 26 0 17 29 21 0 3 0 1 0 2 2 3 1 1 3 2 3 3 27 22 28 35 12 3 2 3 3* 3 3 3 31 41 27 19 12 21 17 32 45 29 43 18 7 1 11 3 3 3 3 3 65 93 105 99 31 3 3 3 3 2 57 42 102 123 75 148 97 112 143 3 32 3 3 1 37 93 128 81 15 2 669A(C) 3 1 115 111 132 121 86 2 Carcinoma B2453 Cholangiocarcinoma 3 Carcinoma 3 81 103 201 85 167 3 with cirrhosis. hepatocellular carcinoma Severe morphological alterations with high degree of hepatocytic dysplasia Nodules of hepatocarcinoma. 832/1 Mild atrophy. 145 . Severe and diffuse fibrosis. Cholangiocellular carcinoma with severe fibrosis. renal epithelium and alveolar and interstitial lung macrophages staining by PB. Mild fibrosis Mild fibrosis Moderate and diffuse form of fibrosis. quails or doves (DORRESTEIN et al. although liver iron. Interesting. PCNA is a 36kD protein that operates as DNA polymerase during phase S of cell cycle and cooperates during DNA synthesis. PCNA and Collagen type IV. In human beings. In view of these activities. ELMBERG et al. 1991).19. In this study. An increase of PCNA expression indicates an high mitotic index rates in hepatocytes population. PB stain and PCNA. the study demonstrates a good correlation between the expression of CEA and hepatocellular carcinoma and CA. cirrhosis and. P53 and AFP expression by hepatocytes. in some cases. to promote the neoplastic transformation of hepatocytes (BACON and BRITTON 1990. was at least 10-fold higher in mynahs than in other species as chickens. present only in hepatocytes (typical finding that characterized primary forms of overload).9 in carcinoma of cholangial origin (HAGLUND et al.9 antigen expression. a product of the p53 gene. as with CEA. we have demonstrated that in mynahs.Statistical analysis revealed a positive good correlation (p<0. These findings suggest that mynahs are a good model for the study of human idiopathic form of haemochromatosis and for evaluations of different factors that induce the progression of the disease into fibrosis. while in cholangiocarcinoma a correlation was observed between PB stain and CA19. in species in which the pathology is described. due to possible increase in the number of transporters when compared with non susceptible species. the high degrees of hepatocellular iron storage correlate positively with the expression of some antigens as mutate form of P53. cirrhosis and hepatocarcinoma. GOSSELIN and KRAMER 1983. For which concern the correlation between PB stain and CEA or CA 19. Calculated values for the uptake kinetics of the probable membrane transporter suggest that mynah bird enterocytes have a significantly higher limiting uptake rate. Finally collagen IV. P53. hepatitis may cause modest elevations of this marker but the marker may still provide the evidence of hepatocellular carcinoma. STYLES 1999). Enterocytes isolated from minahs shows that Fe+++ upatke is much lower than Fe++. an amorphous-non fibrillar form of interstitial collagen that increases during fibrosis and cirrhosis. P53 in normal “wild” form has been rightfully called a “guardian of the genome”. a genetic predisposition is suggested.05) between the following parameters: PB stain and VG/collagen type IV stain. AFP. 2003). the accumulation of iron in the liver may induce fibrosis. 1994. this protein is called in apply emergency brakes when the DNA is damaged by exposure to mutagenic chemicals as iron or free radicals (BACON and BRITTON 1990). 2001).9 expression. 4 DISCUSSION Iron overload is a pathology not frequent in animals but. �-fetoprotein (AFP) is a well-established cancer marker encountered during cancer arising principally in the liver. indicates this transformation directly related to iron overload. acts in the nucleus and has the ability to inhibit the cell cycle. The mynah represents the most commonly reported bird group with susceptibility for iron overload and it was recently shown that the cause of this patho-physiological condition is high uptake and retention of dietary iron (DIERENFELD et al. a positive correlation between PB stain and CEA was observed only in hepatocellular carcinoma . 146 . 183: 11. 5.K.S. Italy Email: giacomo. 125. et al. et al. 121. et al.T.M. The pathology of hepatic iron overload: a free radical mediated process. 124: 26858 – 26868. 7. Doves and Rats. 2. Hemochromatosis: a metabolic disease of softbills. Avian Pathol 2001..S. 6: 1489 – 1496..D.A. Molecular aspects of iron absorption and HFE expression. and ROBERTS P. DIERENFELD E.. STYLES D. 3.M. and BRITTON R.. Rossi. Cancer risk in patients with hereditary hemochromatosis and in their first-degree relatives.5 CITATION INDEX 1. HAGLUND C. Hereditary hemochromatosis. Gastroenterology 2003.R. 30: 479 – 486. BACON B. ELMBERG M.R. Foothill Bird Society 1999.rossi@unicam. EKBOM A.. GOSSELIN S.. Hemosiderosis and dietary iron in birds. 63: 386 – 389. PhD Department of Animal Science. DORRESTEIN G. J Am Vet Med Assoc 1983. LINDGREN J. and SHEPPARD C. DVM.R. METE A. and KRAMER L. 11: 127 – 137.. NIEMELÄ O. J Hepat 2003. 6: 1733 – 1741. and HARRISON S. Difference in tissue expression of tumour markers CA 19. Pathophysiology of excessive iron storage in mynah birds. A comparatives study of iron retention in Mynahs. AUTHORS ADRESS G. Via della circonvallazione 93/95 62014 Matelica MC.J. Proceedings of the Exotic Bird Breeders Symposium... 6. Br J Cancer 1991. 4. HULTCRANTZ R. BACON B. PIOIS M. University of Camerino. 8.J. 86 – 88. 38 14 – 23. Hepatology 1990.. Gastroenterology 2001. BACON B.W. 9. J Clin 1994.J.9 and CA 50 in hepatocellular carcinoma and cholangiocarcinoma. PARKKILA S.it 147 . MARX J. circoviral infection has been reported both in adults (TODD et al. 2001. 2001) and neonates (GOLDSMITH 1995). progressive weight loss. Spain CIRCOVIRUS INFECTION IN A CANARY BREEDING FLOCK J. Circovirus was detected in tissues by DNA in situ hybridization and PCR.Hospital Zoologic Badalona1 SL. PHENIX et al. Circovirus infections are commonly associated with immunodeficiency-related diseases that can be fatal (PHENIX et al. goose circovirus (GCV) and canary circovirus (CaCV) (WOODS and LATIMER 2000. RITCHIE. apathy and nervous signs just before death. pigeon or columbid circovirus (CoCV). Ramis3 DVM. 1 INTRODUCTION Circoviruses are the smallest pathogenic DNA animal viruses that have been described (WOODS and LATIMER 2000. but data regarding clinical signs. 1995). Veterinary practitioner2 Departamento de Medicina y Cirugia Animal3. Grífols1 DVM. Facultat de Veterinària. 2001). F. post-mortem analysis and diagnostic tests results. mortality and morbidity rates and infection progression are scarce. Badalona. Members of the genus Circovirus (family Circoviridae) that infect birds include psittacine beak and feather disease virus (PBFD). D. This paper documents the occurrence of circoviral infection in a canary breeding flock and describes clinical history and signs. Universitat Autònoma de Barcelona. Necropsy and histopathology findings were consistent with immunospression and secondary infections. TODD et al. Barcelona. PhD. The birds died with signs of conjunctivitis. MS.Secondary infections . Some animals died suddenly without previous signs. 2001).Circovirus . Bargalló1 DVM. Dipl ECVP KEYWORDS Canary . In canaries. Perpiñán2 DVM MS.Immunosuppression ABSTRACT Circovirus infection was identified in a breeding canary flock with an outbreak of high morbidity and mortality. diarrhoea. A. 148 . but the owner refused performing more tests. diarrhoea. 2001). lymphocytosis and monocytosis. 3 DISCUSSION Circoviral infection was first reported in canaries in 1995. postmortem examinations and blood work of some live and dead animals. TODD 2000). with goose circovirus (GCV) and canary circovirus (CaCV) being tentatively added to this genus (PHENIX et al. Faecal analyses were negative in 7 live canaries tested. 2001. but histological lesions included hepatitis. apathy. but the virus was not sequenced (TWENTYMAN et al. Birds die of secondary infections. 2001. gallbladder congestion. The breeder had been treating affected animals with metronidazole and reported a slight improvement for some days. and posterior description of infections (TODD et al. The description of GOLDSMITH (1995) of a typical condition for canary circovirus called “Black Spot” should be discarded as these signs are probably produced by secondary infections. 2001). Finally. Canary circovirus seems to induce immunodeficiency by depletion or necrosis of the lymphoid organs. The other bird showed leucocytosis with monocytosis. heteropenia. Results of several PCRs and DNA in situ hybridisation are still going on at the time of writing this manuscript and will be discussed on the presentation. Laboratory studies included fecal analysis. Gross necropsy findings were inconsistent. 149 . canary circovirus should be suspected in cases of mortality affecting several animals. mainly spleen and bursa of Fabricius (GOLDSMITH 1995). Some animals died without previous clinical signs. and pigeon or columbid circovirus (CoCV). necropsy findings. dehydration. 1999). In one animal Pseudomonas aeruginosa and Escherichia coli was culture from kidney. One animal showed severe immunosuppression with leucopenia. 2001). Signs observed varied among individuals and included conjunctivitis. did not report a similar syndrome.2. lymphocytic depletion and focal necrosis in the spleen. Circovirus-like infection has also been describe in a southern black-backed gull (Larus dominicanus). sequenced and named as canary circovirus (CaCV) (PHENIX et al. TODD et al. Currently. Subsequently this virus have been cloned. Circovirus infections are commonly associated with immunodeficiency-related diseases that are potentially fatal (PHENIX et al. psittacine beak and feather disease virus (BFDV). and nervous signs before death. as a condition called “Black Spot” disease (GOLDSMITH 1995). including the present paper. lung. and failure to thrive (GOLDSMITH 1995). so clinical signs. This condition included signs like abdominal enlargement. progressive weight loss. DNA in situ hybridization was positive for Circovirus in one of two animals. especially when histopathology shows depletion or necrosis of the lymphoid organs. the genus Circovirus of the family Circoviridae comprises porcine circoviruses types 1 and 2 (PCV1 and PCV2). and liver. CASE REPORT In August 2003 a canary breeder reported high mortality and morbidity in his breeding flock. and morbidity and mortality can show a wide range of variation depending on the agent or agents involved. and PCR was positive for Circovirus in other two of two animals. 2001. Blood analyses were performed in 2 live birds with different results. Mortality continued for two months. after which mortality remained high. TODD et al. purulent meningitis. Lake Worth: Wingers Publishing Inc. 4. Circovirus-like infection in a southern black-backed gull (Larus dominicanus). 82: 2805 . J Gen Virol 2001. Avian Pathol 2001.163. 2.350. PHENIX KV. 08912 Badalona (Barcelona). Nucleotide sequence analysis of a novel circovirus of canaries and its relationship to other members of the genus Circovirus of the family Circoviridae. 30: 321 . Spain Email: jordi. WESTON J.325.394. et al. Proc Assoc Avian Vet.516. Avian Pathol 28: 513 . TWENTYMAN CM. Avian Viruses: Function and Control. WESTON JH. Documentation of passerine circoviral infection. 5. WOODS LW and LATIMER. Circovirus infection of nonpsittacine birds. AUTHORS ADDRESS Jordi Grífols Zoològic Badalona Veterinària Conquesta 74.es 150 .grifols@hzb. ALLEY MR. 1995. 3.4 REFERENCES 1. YPELAAR I. 14(3): 54 . Avian Pathol 29: 373 . J Avian Med Surg 2000. Nucleotide sequence-based identification of a novel circovirus of canaries.2809. RITCHIE BW. et al. MEERS J. 7. 6. Nashville 1995. TODD D. GOLDSMITH TL. Circoviruses: immunosuppressive threats to avian species: a review. BALL NW. et al. TODD D. KS. 349 . 91µg/l and 123µg/l. Lierz KEYWORDS Vitamin B . Carnarius. Germany. continued by an oral gavage of 2mg/kg twice daily. a 45 day old female goshawk was admitted with irregular attacks of central nervous signs like opisthotonus.Thiamine deficiency – Beriberi – Nutrition – Raptors . Therefore a continuous therapy of 2mg/kg thiamine hydrochloride once daily for a few days and a change of the diet are recommended as treatment. 6 healthy captive bred goshawks at the age of 45-55 days showed an average of 113µg/l (95-120µg/l).Birds of prey Goshawk ABSTRACT In the last 2 years an increased number of juvenile goshawks (Accipiter gentilis) with central nervous symptoms were presented to the Institute of Poultry Diseases of the Free University Berlin. Two healthy free ranging juvenile goshawks showed a level of 133µg/l and 162 µg/l and 3 starved free ranging juvenile goshawks showed results of 51µg/l. The last case. rotations and somersaults especially when the bird was handled or stressed. As reference values for thiamine level in goshawks are not presented in the available literature.Free University of Berlin. The therapy of the bird with the suspected thiamine deficiency was an initial intramuscular injection of 4 mg/kg thiamine hydrochloride. A study was performed to establish comparable values for goshawks. 4 adult captive goshawks at the age between 10-23 years showed an average of 97. 14163 Berlin Germany CENTRAL NERVOUS SYMPTOMS CAUSED BY THIAMINE DEFICIENCY IN JUVENILE GOSHAWKS (ACCIPITER GENTILIS) M. Already after the first injection of thiamine the symptoms resolved. M.25µg/l (44-149µg/l). As a diagnosis a thiamine deficiency was suspected. Hafez. H.2µg/l in plasma while all other blood values examined revealed normal results. A. In spite of these central nervous attacks the bird showed a normal appetite. 151 . M. Faculty of Veterinary Medicine Institute of Poultry Diseases Königsweg 63. Henning. Henning. Comparable to the goshawks presented before the bird always preened after an attack. The bird demonstrated a thiamine content of 2. After 5 days treatment the blood value of thiamine raised to 476µg/l. The goshawk was wild caught at the age of approximately 20 days and received a diet of thawed day-old chicks. The birds were between 45-55 days old and were fed with thawed day-old chicks. H. J. above all chickens. legumes. consequently it is required in their diets (VOET and VOET 1995). quails contain about 0. thiamine deficiency may occur. In chicken. Birds. OLKOWSKI and CLASSEN (1996) explored in chickens the thiamine status in blood and tissues in response to a wide range of dietary supplementation of thiamine. for example. A thiamine deficiency can arise from decreased intake or absorption. yeasts. and maintaining memory (HALLIDAY et al. 2000). Thiamine is found in nearly every animal and herbal aliment but generally only in few amounts. Thiamine (vitamin B1) is mainly existent in its active form thiamine pyrophosphate (TPP. In birds fed large amounts of raw fish or fish that has had an opportunity to sit without inactivation of the enzyme thiaminase. At the moment there are only a few studies about the required intake of thiamine in birds. in liver. for example the species of fish fed (ROUDYBUSH 1997).30-0. This vitamin is a factor in the normal intestinal function and in the health of cardiovascular and nervous systems (WORLD HEALTH ORGANIZATION 1999).1 INTRODUCTION Thiamine is a water soluble and heat-labile vitamin out of the group of B vitamins. It is also a coenzyme of the transketolase. growth and hyperthyroidism (WORLD HEALTH ORGANIZATION 1999). also called thiamine diphosphate. histologically the deficiency of vitamin B1 demonstrates a polyneuropathy 152 . turkeys and quails.36 µmol (0. heart and kidney. It is involved in the synthesis of collagen. 1996). On the other hand juvenile rats contain about 0. for example in the citric acid cycle.33mg/100g and mice 0. 1980). were often used to study the pathogenesis and the symptoms of beriberi for human medicine (DJOENAIDI et al. Furthermore a thiamine deficiency can occur from an increased amount of enzymes that require thiamine or from interference by other ingredients of nutrition. The probability of a reduction in thiamine as a result of thiaminase activity is dependent upon a number of factors. from increased turnover or from loss of thiamine by enteritis or coccidiosis (MANORE 2000).18 mg/100g. adult rats 1. The thiamine pyrophosphate is an essential cofactor of decarboxylation reactions in many metabolic pathways. 2000) or tannin (BEGOVIC et al. improving cognitive function. 1995) as well as the histological signs of thiamine deficiency (ANJALI et al.02mg thiamine per 100g (FORBES and FLINT 2000). Day-old chicks are commonly used as feed in captive raptors. Furthermore TPP plays a role in the release of the neurotransmitter acetylcholine (PARKHOMENKO et al. Whole day-old chicks contain an average of 0. 1994). Thiamine is neither synthesized nor stored in significant amounts by the tissues of most vertebrates.6 µmol (1.7 mg) thiamine per day or 0.31mg/100g. The requirements of thiamine are increased when carbohydrates are taken in large amounts and during periods of increased metabolism. BRUE (1994) state a recommended allowance of thiamine for maintenance about 5ppm per day for most psittacines and the commonly kept passerines. In comparison. fever. Blood thiamine concentration in those birds supplemented with 8 mg/kg body weight thiamine increased at day 7 and remained relatively constant.12 mg) thiamine per 1000kJ and day (LÖFFLER 1998).065mg thiamine per 100 g.10-0. for example aflatoxin (VOIGT et al. muscular activity. VOET 1995). 1980). The highest concentrations of thiamine are found in un-grinded cereals. A healthy adult human being needs an intake of 5. ThDP) (VOET. or by drugs like amprolium (ANJALI et al. Because of this improving a thiamine deficiency was suspected. coli.with axonal degeneration. Typically the birds showed these disorders at the age of 45-55 days and were fed with thawed day-old chicks. Furthermore the diet of this peregrine falcon was changed from young cockerels into whole pigeons and quails. In addition levamisole and niclosamide base were applied orally followed by a vitamin and mineral complement added to the feed. Chlamydophila psittacii. They were fed with dead sheep (including viscera). the decrease of the enzyme transketolase in the erythrocytes is used. the neck stretched out and the beak half open. PERICARD and ANDRAL (1993) described several cases of nervous disorders called “staggers” in free ranging rescued griffon vultures (Gyps fulvus) of South Central France and the French Pyrenees. a pathogenic E. As the birds improved after a thiamine supplementation a thiamine deficiency was suspected. seizures and death. for one to three minutes. After all in none of the cases the diagnosis was confirmed. These signs were consistent with the polyneurotic signs observed in poultry (ROUDYBUSH 1997). 1972). 2000). Free ranging honey-eaters in urban areas of southern Australia may develop thiamine deficiency during winter (MACWHITER 1994). a thiamine deficiency leads to a degeneration of the cells lining the duodenal crypts of Lieberkuhn. Case report In the last 2 years an increased number of juvenile goshawks (Accipiter gentilis) with central nervous symptoms were presented to the Institute of Poultry Diseases of the Free University Berlin. For example. Salmonella. 2000). During one year 11 birds died. Reports about a suspected thiamine deficiency in birds of prey are rare and un-detailed. The therapy 153 . Consequently the pentose phosphates in the erythrocytes are increased. minimal segmental demyelination (DJOENAIDI et al. WARD already described in 1971 a case of a peregrine falcon with opisthotonus. In psittacines a thiamine deficiency may lead to loss of appetite. Nematodes and viral encephalitis were found in some of the birds respectively. 1995). The present paper describes central nervous symptoms in juvenile goshawks due to thiamine deficiency and laboratory approach to determine the plasma thiamine level in goshawks. congestion and oedema of brain (ANJALI et al. one wing half extended. For a diagnosis of a thiamine deficiency in human medicine. opisthotonus. with dilatation and filling of the crypts with cellular debris and necrotic cells. Moreover degeneration of testes and ovaries and atrophy of liver can be observed (ANJALI et al. In a single case of feeding polished rice to adult cockatiels. Hereupon the affected vultures improved. The only treatment the remaining birds got was an injection of vitamin B1 and vitamin B6. He suspected a thiamine deficiency after a long period of unsuccessful treatments and a sudden improvement of the symptoms after a treatment with 2 days primidone (125 mg once daily) and 5 days intramuscular injections of thiamine hydrochloride (1 mg/day). This can be measured before clinical symptoms appear (LÖFFLER 1998). But a deficiency of thiamine is uncommon in birds on a seed diet because seeds and grains generelly contain a sufficient amount of thiamine. head tremors occurred before the experiment was discontinued. The affected birds turned round quickly. Furthermore. and vacuolation of the pancreatic acinar cells with hyaline body formation (GRIES et al. Germany. The plasma was examined for aspartate aminotransferase. The present case. 5 juvenile goshawks of both genders were found in the wild and brought to the institute by private persons. total protein. calcium. albumin. similar to the birds presented before. uric acid. pigeons and thawed day-old chicks. For a clear diagnosis of a thiamine deficiency comparable reference values of thiamine in blood are missing. A blood sample was taken from every bird from the V. bile acids. lactate dehydrogenase. triphosphate derivates of thiamine and the free thiamine) is oxidised into bluish coloured thiochrome and detected with HPLC/fluorescence method (HPLC = High Performance Liquid Chromatography). 154 . First proteins were denatured and afterwards the enzymatic hydrolysis is accomplished with acidic phosphatase. This bird was wild caught at the age of approximately 20 days and fed with thawed dayold chicks. total protein. All birds received a detailed clinical examination. a 45-days-old. di-. neurological and radiographic examination a thiamine deficiency was suspected due to the experience with the goshawks presented before. phosphate. 2 MATERIAL AND METHODS In total 11 juvenile and 4 adult goshawks were used to determine the level of vitamin B1 in blood-plasma. The whole vitamin B1 (consists of mono-. creatine kinase. After 5 days treatment the blood value of thiamine raised to 476µg/l. albumin.of such birds was an initial intramuscular injection of 4 mg/kg thiamine hydrochloride. after blood sampling. The blood was centrifuged 5 minutes. an endoscopic examination. 5000 U/min. Six juvenile birds of both genders were captive bred from a private breeder and at the time of blood sampling 45-55 days old. Therefore a continuous therapy of 2mg/kg thiamine hydrochloride once daily for a few days and a change of the diet are recommended as treatment. lactate dehydrogenase. cholinesterase. They were fed with a mixed diet of fresh killed rats. continued by an oral gavage of 2mg/kg twice daily. and the birds from the wild in addition a radiological and. The 4 adult goshawks came from the same breeder and they receive only thawed day-old chicks. Already after the first injection of thiamine the symptoms resolved. calcium. After a general. Immediately after such an attack the bird started to preen and showed a normal appetite. Typically this goshawk showed attacks with central nervous symptoms when the bird was handled or stressed. cholinesterase. 2 birds were generally healthy. creatine kinase. metatarsalis plantaris superficialis medialis into a Lithium-Heparin tube. bile acids. For testing thiamine values a sample of 250 µl from EDTA plasma is used. found freshly injured. Therefore it was tried to establish those. uric acid. potassium and thiamine. phosphate. 5 µm as well as citrate buffer-methanol-alloy (VET MED LABOR 2001). Thiamine level was 2. female goshawk. 3 birds were starved. Blood was taken and tested for aspartate aminotransferase. showed severe central nervous disorders with opisthotonus and rotations. The HPLC analysis is carried through a HPLC system LC 6 apparatus via Lichrosphere 100 C18-columns. potassium and thiamine.2µg/l while all other blood values were within reference ranges. starved 6 m 7 f 8 f 9 m 10 f 11 m 12 m 13 m 14 f 15 f . healthy Captive bred.0 Captive bred. healthy Captive bred.0 120. 155 Free ranging. healthy Free ranging.0 90. healthy Captive bred. healthy Captive bred.0 Free ranging.0 133. anaemia 51. healthy Captive bred.0 91. Results of the plasma thiamine content of 15 goshawks in comparison to a case with CNS symptoms and a suspected thiamine deficiency. healthy Captive bred.0 162. healthy 114.0 116.0 Captive bred.3 RESULTS Table 1. healthy 116.0 95. starved Free ranging.0 44.2 CNS disorders 50 days 50 days 55 days 55 days 55 days 60 days 5 years 10 years 15 years 20 years 70 days 75 days 65 days 70 days 75 days Captive bred. healthy 117.0 149. starved.0 106. Case f 45 days 2.0 123.0 1 m 2 m 3 f 4 f 5 f The examination of all above mentioned blood parameters (except thiamine) revealed results within usual ranges. humerus fracture. Thiamine in plasma Goshawk No Sex Age Characteristics (µg/l) Wild caught at the age of 20 days. The obtained results of thiamine and of the clinical examination are presented in table 1. healthy Free ranging. healthy Captive bred. Seeing the regular onset of such disorders in juvenile goshawks around 45-55 days on a thawed day-old chicks diet it is suggested that the feed and its thiamine content plays an important role at the development of a thiamine deficiency. As all examinations revealed no pathological findings a vitamin B1 deficiency was suspected. indicates a thiamine deficiency. The blood thiamine value (2. interestingly the two free ranging and healthy juvenile goshawks showed thiamine contents of 133µg/l and 162µg/l. The inhomogeneous constitution of the tested group of birds. Even one of the starved goshawks showed a result of 51µg/l thiamine in plasma and showed no clinical symptoms of a thiamine deficiency. juvenile but starved goshawks had thiamine values in blood of 51µg/l. It might 156 . Also the TPP content in the heart increased in response to both maternal and offspring supplementation. 91µg/l and 123µg/l. Additionally the thiamine supplementation in the offspring diets increased blood TPP (OLKOWSKI. Moreover this method has not been used for birds so far. FORBES and FLINT (2000) showed that the vitamin B1 content in day-old chicks is not as high as in other diets such as rats. Even the three free ranging.5µg/l and therefore much higher than the value of the presented bird. Based on the results of the 12 healthy birds the average of the plasma thiamine values is 113. Therefore the given values are just for orientation and reference values have to be established in further studies. Another proof of the deficiency of vitamin B1 in the presented bird is the successful therapy with thiamine hydrochloride only. Maybe these day-old chicks did not have enough amount of thiamine because of the inadequate supply of thiamine to the parents.2µg/l. The six juvenile. captive bred and healthy goshawks showed thiamine contents in blood between 95-120 µg/l. Additionally only 15 goshawks were tested for thiamine in plasma in this study. Furthermore the kind of freezing and the storage time can also have an effect on the thiamine content of day-old chicks. CLASSEN 1999).2µg/l) of the presented bird was much lower compared to the other juvenile goshawks tested. The bird had a thiamine content in plasma of 2. It was found that maternal supplementation of thiamine increased content in the heart of the offsprings. Freezing is a drying process and long-term storage (unless sealed) can reduce the water content of feed. particularly in terms of water soluble vitamins and vitamin E. juvenile and adult ones. Determination of thiamine blood values is easy and appropriate to a diagnosis of thiamine deficiency. received different feed. does not allow the establishment of statistical proved reference values for thiamine.4 DISCUSSION A juvenile goshawk was presented with central nervous symptoms. Therefore other goshawks were used to detect the usual range of thiamine in plasma for comparing with the presented case. This especially due to the experience that similar symptoms in juvenile goshawks were observed in the past and responded to thiamine supplementation. As reference values for vitamin B1 in goshawks are missing a deficiency was not proved in this case. Food kept for protracted periods in domestic and commercial freezers deteriorates in nutritional quality. Additionally this method is easier than the one used in human medicine where not the thiamine in plasma or serum but the increased amount of pentose phosphates in the erythrocytes were measured. That’s why feed should not be kept stored frozen for more than 3 months (FORBES and FLINT 2000). 177 .1285. 157 . BEGOVIC S et al. BRUE RN. 6.24. 4. Sequential pathological changes in experimental thiamine deficiency in Japanese quail (Coturnix coturnix japonica). Experimentally induced beriberi polyneuropathy in chickens. FORBES NA and FLINT CG. 71. 5 CITATION INDEX 1.30. It is suggested that the high incidence of thiamine deficiency in young birds compared to adults is connected with their increased requirement of thiamine because of their growth. Veterinaria. Goshawks present CNS symptoms in a typical way.also play a role that partial defrosting of the day-old chicks during transport from the breeder to a dealer and to the falconers is responsible for their reduced vitamin B1 content. 29: 1 . 2: 245 . HARRISON GJ and HARRISON LR (ed): Avian Medicine: Principles and Application. 35(1): 53 . Inc. Based on all these facts a pure diet with thawed day-old chicks cannot be recommended for goshawks. Neuropathological correlates of memory dysfunction in the Wernicke-Korsakoff syndrome. On the other hand vitamin B1 as a water soluble vitamin can quickly be excreted and overdose might not be a big problem. Slight clinical symptoms might resolve by changing the diet. This excessive high level in the blood value shows that this thiamine application rate was overdone.179. Therefore half of the dose should be far enough but further studies are necessary to establish a recommendable treatment. Effects of tannin and pyrogallol on the course of experimental B-avitaminosis in fowls. 29 . 7. Lake Worth: Wingers Publishing. Also it can be possible that the enteral enzymes of juvenile birds are not yet fully developed. As therapy for the presented bird it was given an initial intramuscular injection of 4mg/kg thiamine hydrochloride. continued by an oral gavage of 2mg/kg twice daily. DJOENAIDI W et al. rotations and somersaults above all when the goshawks were handled or stressed. The pathology of thiamine. 5. 102(10): 1269 . 3. pantothenic acid and niacin deficiencies in the chick. Alcohol Alcohol Suppl 1994. It is conspicuous that the diseased birds always showed a normal appetite and preened after such an attack with central nervous disorders. HALLIDAY G.251. According to the available literature it is the first time that a vitamin B1 deficiency in goshawks was proven and blood values of vitamin B1 for orientation were provided. ANJALI et al. CULLEN K and HARDING A. 23 . 24:1. They show irregular attacks with opisthotonus. 2. Raptor Nutrition.18. Electromyogr Clin Neurophysiol 1995. So the enzymes cannot convert enough TPP into thiamine which can be absorbed from the intestine.2. but this seems not very likely. with particular reference to nervous disorders. J Nutrit 1972. Indian J Vet Path 2000. Nutrition. In RITCHIE BW. GRIES CL et al. After 5 days of treatment the blood value of thiamine increased to 476µg/l. 16 . Honeybrook Farm Animal Foods 2000.60. 1994. Yugoslavia 1980. Rodents (above all rats) including their livers seem to be more adequate at least as part of the diet. 651. In LÖFFLER G and PETRIDES PE. Faculty of Veterinary Medicine. 856. 10. 664 . Germany Email: gefluegelkrankheiten@vetmed. LÖFFLER G. HARRISON GJ and HARRISON LR (editors). Berlin: Springer-Verlag 1998. Malnutrition. Inc. Avian Medicine: Principles and Application. Stoffwechsel der Kohlenhydrate. In Löffler G. Vitamine. Lake Worth: Wingers Publishing. The full list of references is available from the author AUTHORS ADRESS M. 6th edition. MACWHITER P. Institute of Poultry Diseases. LÖFFLER G. 6th edition. 14163 Berlin. Biochemie und Pathobiochemie.de 158 . Carnarius Free University of Berlin. Königsweg 63. Petrides PE. In RITCHIE BW.. 649 .fu-berlin. 1994. :387-9 9. Biochemie und Pathobiochemie.8. Berlin: Springer-Verlag 1998.665. University of Leipzig.E. Although first observed more than two decades ago. Raue2 and M-E.YPD . Clinical. In order to collect more convincing data. The results of this comprehensive study indicate that YPD is a multifactorial disease in which PiCV might be a crucial factor. The aetiological agent remained unknown. In contrast to other viral pathogens. pathological. the etiological agent of this disease still remained unknown. Faculty of Veterinary Medicine. Pigeons in their 4th to 12th week of life exhibited those clinical signs more frequently and more severely than pigeons of other ages.Pathology . University of Leipzig. Intranuclear inclusion bodies were observed in various organs especially in the bursa of Fabricius.Clinic for Birds and Reptiles1. the presence of the genome of pigeon circovirus (PiCV) was demonstrated in lymphoid tissues from all pigeons with YPD. mainly Germany. coli ABSTRACT A disease in young racing pigeons. YOUNG PIGEON DISEASE V. Institute for Virology2. 159 . Numerous pathological and histopathological findings were observed. haematological.Immunosuppression . Krautwald-Junghanns1 KEYWORDS Young pigeon disease . Department of Small Animal Medicine. 1 INTRODUCTION A disease in young racing pigeons associated with high morbidity and mortality rates reported from parts of Central Europe. Schmidt1. was designated as young pigeon disease (YPD) by pigeon breeders. Faculty of Veterinary Medicine. parasitological and microbiological examinations were performed. R. a comprehensive study was performed with pigeons present in German lofts with or without clinical outbreaks of YPD. histopathological. associated with high morbidity and mortality rates. was designated as young pigeon disease (YPD) or young bird sickness. designations such as “swollen gut syndrome” describing pathological observations were also used by pigeon fanciers and veterinarians. most probably caused by the multitude of pathogens which were isolated. Germany A COMPREHENSIVE STUDY ON A DISEASE COMPLEX ASSOCIATED WITH PIGEON CIRCOVIRUS INFECTION. possibly by inducing immunosuppression in infected birds.PiCV . and all lofts were tested negative for salmonellosis. intramuscularly) and diazepam (5mg/kg body weight.600 questionnaires revealed that pigeons were affected by YPD mainly between 4 to 12 weeks post weaning. the type of the pigeon loft. depression. grown up in 4 lofts without YPD. however. a second part of this study was performed thereafter including examinations for Chlamydophila spp. ruffled feathers. 160 . Various non-infectious causes as well as infectious pathogens. In individual cases. 2 MATERIALS AND METHODS Seventy six juvenile racing pigeons of both sexes obtained from 29 lofts located in various parts of Germany were investigated. Clinical signs observed were unspecific: anorexia. polyuria and fluid filled crop. DE HERDT and VAN GINNEKEN 1994). intravenously) after an initial anaesthesia with ketamine (40mg/kg body weight. a questionnaire was developed and distributed to German breeders of racing pigeons. such as Spironucleus columbae. It was reported that generally less than 20% of the young pigeons were affected and that mortality rates were about 20%.In order to get more insight into the epidemiological situation of YPD. Due to the results gained. Escherichia coli (E. Out of these. All pigeons hatched in 2003 or 2004. the occurrence of YPD and the clinical signs observed. intramuscularly). the feeding and drinking habits. Pigeons were humanely killed with potassium chloride (2mmol/kg body weight. Furthermore. a comprehensive study was performed with pigeons present in 25 German pigeon lofts with clinical outbreaks of YPD in 2003 or 2004 compared with pigeons of 4 lofts without clinical outbreak of YPD in 2004. were included in the study. were considered to contribute to the pathogenesis of YPD (DORRESTEIN and VAN DER HAGE 1992. Various clinical and pathological pictures for YPD have been described. the birds were necropsied according to standard procedures (LATIMER and RAKICH 1994). 67 pigeons were from 25 lofts with clinical outbreaks of YPD diagnosed by veterinarians according to the following criteria: (i) increased mortality rates among pigeons in their 3rd to 20th week of life. For comparison 9 young racing pigeons. (ii) poor racing performance. A detailed history of each pigeon’s husbandry and the course of the disease were reported by the owners. diarrhoea. The analysis of more than 1. Among others. coli) or avian adenoviruses. parasitological and microbiological examinations. questions referred to the number of pigeons. mortality rates of more than 50% were also reported. Clinical examinations including white blood cell counts (WBC) and differentiation of white blood cells were performed and evaluated according to standard procedures and compared to describe reference haematological values for racing pigeons (VOGEL 1992. it became evident that stress conditions such as long transports or hot weather during the young pigeon races might play an important role in the induction of YPD. vomiting. and viral agents. The aim of this study was to get knowledge on pathological and histological alterations in pigeons with YPD and to compare those with the results gained from haematological. In order to collect more convincing data. RUPIPER and EHRENBERG 1994). testes or ovary and bone marrow were collected under sterile conditions. torticollis (5. In the case of WBC. Ingesta from small and large intestines were mixed with saturated sodium chloride solution at a 1:1 ratio and floated for 20 minutes. swabs were taken from liver. Germany) in an agglutination test as described. small and large intestines. Wesel. heart. Wesel. ventriculus. heart. spleen. USA). Deviations from normal findings were verified for significance using cross tables and Pearson´s Chi2 test. embedded in paraffin wax and sectioned at 4µm for light microscopy examinations. anorexia. Germany) as well as anaerobic conditions (AnaerocultA. bone marrow. Germany) was inoculated and incubated at 38°C for 72-120h. 10 pigeons from affected lofts died during transport.5. lung. Suspicious colonies were screened with polyvalent antibody of Salmonella A-E (Enterocolon anti-Salmonella I (A-E). Resistance screening of the isolated bacteria by agar diffusion test and determination of the minimum inhibitory concentration ensued. Germany). Formalinfixed samples were dehydrated. A semi-quantitative evaluation of 5 microscopic fields was performed at 200x magnification. Berlin. version 11. For mycological examinations. microaerobic (AnaerocultC. Germany) at 38°C for 24h. cachexia. 6 out of 57 pigeons). Samples from crop and small and large intestines were collected immediately after opening of the carcasses and investigated microscopically for flagellates using 200x and 400x magnification. Dade Behring. Oxoid.8%. mean values and standard deviations were also calculated. Sabouraud’s dextrose agar (Oxoid. SIFIN. crop. 3 RESULTS Most of the pigeons (63 out of 76) were sent to examination alive and during the hot summer month (14 out of 29 lofts). 3 out of 57 pigeons). Cultivation of bacteria was performed at 38°C under aerobic. Drop volumes taken from these media were incubated on selective culture plates (XLT4 agar. Wesel. Parts of these organs were fixed in 10% neutral buffered formalin for at least 24h. In 32 out of 57 pigeons (56%) from YPD-affected lofts clinical signs were observed. small and large intestines. samples taken from liver and the contents of gut were cultivated in selenite lactose medium (Oxoid. vomitus. respectively. Merck. Statistical analysis was performed using the program SPSS.8% each. spleen. Germany) at 100x magnification using oil. parathyroid.. Chicago. brain. respectively. Germany). Merck.3%. All sections were stained with haematoxylin and eosin and evaluated. 1 out of 57 pigeons). kidneys. Wesel. 21 out of 57 pigeons).Impression smears prepared from liver. Germany) for 24 and 72h. Clinical signs were observed in none of the pigeons from lofts without 161 . dyspnoea and splayed leg (1. proventriculus.5% each. Darmstadt. polyuria. small and large intestines were examined cytological (DiffQuik. IL. Liver. using Columbia agar supplemented with defibrinated sheep blood and brilliant green agar (Oxoid. Germany) at 38°C for 48h.1 (SPSS Inc. thymus. lung. Heidelberg. Marburg. bursa of Fabricius. Differentiation of bacteria was carried out using Chrystal™ Tube (BD. kidneys. Darmstadt. yellow urates (10. lung. Clinical signs were greenish to black diarrhoea (36. For bacteriological and mycological investigation. For detection of salmonella. adrenal gland. thyroid. 9%) nutritional condition. Enteritis was found more often in pigeons who were dissected in summer (p<0. Multiglobular basophilic intranuclear and intracytoplasmatic inclusion bodies were observed in lymphocytes and macrophages of the bursa of Fabricius in 21 of 57 (36.5%).001) than in birds without clinical signs. but were more than 12 weeks old. Pigeons with fluid filled small intestine (p<0.8%) pigeons from lofts with YPD. 26 out of 57 pigeons. and bursa of Fabricius (1 out of 67 pigeons.001) or inclusion bodies in the bursa of Fabricius (p<0. In contrast. 16. 162 . Many of the dissected birds from lofts with signs of YPD (n = 67) showed a poor (34.2%). they were present in lymphocytes and macrophages of the gut-associated lymphoid tissue of 3 pigeons. In none of the pigeons cytological alterations such as necrosis or inclusion bodies were detected in smears taken from the bone marrow.5±0.087). Furthermore. In small and large intestines.4%) pigeons and in the bursa of Fabricius of 9 out of 57 (15.57±0.9%) as well as 2 out of 9 pigeons without YPD (22. Yellow fluid in small intestine was found in 18 out of 67 pigeons with YPD (26.6%.66x103/µl]a). [8. In contrast.3%) to bad (17. respectively. p<0. Pigeons in their 4th to 12th week of life exhibited the signs mentioned above more frequently and more severely (17 out of 23 pigeons representing 73.5%) were seen only in pigeons from lofts with signs of YPD. mild acute catarrhalicpurulent enteritis (3 out of 67 pigeons.0%).4%).4%). Clinical signs were observed more often in pigeons who were examined in summer (45. kidneys or spleen. 6. green fluid in crop.033) and in pigeons with depletion of spleen lymphocytes (p<0. In none of the pigeons histological alterations such as necrosis or inclusion bodies were detected in the bone marrow. liver and spleen (11 out of 67 pigeons each. [8.5%). none of the pigeons from YPD-free lofts showed haemosiderosis in liver.008). of the bronchus-associated lymphoid tissue and of the spleen in one case each as well as of the liver of 5 out of these 67 pigeons. 1.8%) pigeons. Lymphocytic depletion and lymphocellular necrosis was observed in the spleen of 13 out of 67 (19.0%). severe multifocal granulomatous enteritis (4 out of 67 pigeons. Pigeons showing leucocytosis (8 out of 50 pigeons) originated from lofts with YPD. Gross lesions of lung and air sacs (12 out of 67 pigeons.035).0%). Multiglobular basophilic inclusion bodies were also found in the bursa of Fabricius of one pigeon without YPD.9%). 17.053). 4. in pigeons with clinical signs of YPD (p<0. proventriculus and ventriculus (p<0. 10. Other histological changes were induced by septicaemia or mold-infection.9%) than pigeons of other ages.085). and no inclusion bodies were detected in erythrocytes or leukocytes of blood smears. kidney (7 out of 67 pigeons.history of YPD. 9.079) were affected by haemosiderosis significantly more often. Blood smears of 50 out of 67 pigeons from lofts with YPD and 6 out of 9 pigeons from lofts without YPD were evaluated. Inclusion bodies were observed with a statistically significant higher incidence in pigeons with fluid filled small intestine (p<0.69x103/µl]). of the kidney of 2 pigeons. Leucopenia was seen in 7 out of 50 pigeons from lofts with YPD (14%. heart and pancreas (3 out of 67 pigeons each. proventriculus and ventriculus had a significantly higher prevalence in diseased birds (p<0. histological changes were grouped as follows: mild to severe catarrhalic duodenitis (6 out of 67 pigeons. intestines (4 out of 67 pigeons. Leucopenia was seen in 3 out of 6 pigeons from lofts without YPD (50%. 6. Findings of greenish liquid in crop. whereas pigeons from YPD-free lofts were of good nutritional status. 4. 7% of the pigeons.02) were isolated more frequently from pigeons with inclusion bodies in the bursa of Fabricius. facultative pathogenic E. pigeons from YPD free lofts (8 out of 9 pigeons. In 10 out of 67 pigeons with E.019).5%) pigeons. and 19 out of 76 (25.017) as well as molds (p<0. 67. spleen. Spironucleus columbae was present in the intestines of 15 out of 76 (19. were demonstrated in 3 out of 76 pigeons (3. Trichomonas gallinae affected 11 out of 76 (14.051). from both. Yeast or molds were only detected in pigeons with YPD. 30 out of 76 (39. Having this in mind. Spironucleus columbae. greenish liquid in crop.5%) pigeons carried pigeon lice (Columbicola columbae columbae). proventriculus and ventriculus (p<0. Intranuclear inclusion bodies as observed here are mostly caused by DNA viruses.8%). no salmonella were isolated from liver and intestines of all pigeons. SHIVAPRASAD 1994).0%) pigeons feather mites (Falculifer rostratus. in pigeons with clinical signs (32 pigeons) E. was observed in only 19. coli isolation from the large intestine only (21.019). These assays allowed the detection of viruses such as PiCV which. Interestingly. However.As expected.9%) and from those with YPD (45 out of 67 pigeons. numerous pathological and histopathological findings were observed. This hypothesis is supported by the observation that lymphoid depletion and lymphocellular necrosis were observed in spleen and bursa of Fabricius and that intranuclear and intracytoplasmatic inclusion bodies were demonstrated most frequently in the bursa of Fabricius but also in lymphocytes and macrophages of the liver. E.9%. coli was isolated from 69. YPD–free lofts and lofts with YPD. Eimeria spp.05) and fluid filled small intestine (p<0. PCR protocols for the detection of pigeon DNA viruses were developed. at present.) were observed in 9 out of 76 birds (11. it was detected statistically more frequently in pigeons with clinical signs (p<0. coli was isolated more often from small intestine additionally (59.3%. Megnina columbae).7% (53 out of 76) of the examined pigeons.7%) pigeons. coli isolated from 53 of 67 pigeons affected with YPD and other facultative pathogens were identified in the majority of cases. 4 DISCUSSION YPD is characterised by unspecific clinical signs and pigeons are affected in their 4th to 12th week of life more severely and frequently. However. In contrast to former studies (DORRESTEIN and VAN DER HAGE 1992) 163 . previously discussed as the causative agent. 7 out of 32 pigeons). Furthermore.9%) and helminths (Ascaridia spp. and Capillaria spp.008). kidneys and lung. Although E. As expected. indicating that the immune system of these pigeons might be affected. intestines. numerous bacteria were isolated from pigeons with YPD and without YPD. Comparable results were obtained from the clinical examination in this study. Statistical significance for the presence of this parasite was proven for pigeons with clinical signs (p<0. can not be isolated in tissue culture systems (WOODS and SHIVAPRASAD 1997). most certainly caused by numerous (secondary) pathogens. 19 out of 32 pigeons) contrary to E. Recently such inclusion bodies have been reported from the bursa of Fabricius of pigeons infected with the pigeon circovirus (PiCV) (WOODS 1994. coli was observed in both. However. Main site of isolation was the large intestine. In these yeast (p<0. 88. coli isolation from the small intestine.2%). However. the above mentioned histopathological alterations of the gut were detected (p<0. Consequently. The multitude of clinical signs observed during an outbreak of YPD reflects these various secondary infections. Attempts to demonstrate the presence of avian polyomavirus remained negative as well. these pigeons might represent persistently infected carriers and might be responsible for the spread of this virus. coli was isolated more often from the small intestine of clinical ill pigeons. a facultative pathogenic bacterium belonging to the normal flora of the large intestine. Taken these results into account. In particular E. However. The exposure of such pigeons to stress conditions such as racing. overcrowding. YPD represents a multifactorial disease in young pigeons. the low number of pigeon herpesvirus positive results (15. resulting in septicaemia. This hypothesis is supported by the fact that pigeons with YPD are highly susceptible to secondary infections with bacteria. together with published data.6%) revealed the expected prevalence rate (EF KALETA. These results. most of the young racing pigeons with YPD did not show leucopenia. hot weather. personal communication) and indicates that the immunesystem might be able to be reactivated. 164 . From the results of this comprehensive study it was predicted that most probably. 5 ACKNOWLEDGEMENT This study was supported by the Verband Deutscher Brieftaubenzüchter e. PiCV DNA can be detected in various organs and blood in different amounts. the induction of systemic infections. PiCV DNA was demonstrated in all pigeons with YPD. This irregular gut flora supported the spread of parasites and other secondary pathogens. were most frequently tested positive.V. Furthermore E.8%) indicating that they were viraemic. In pigeons with YPD neither necrosis nor inclusion bodies were observed in the bone marrow. The high quantity of E. Lymphatic tissues. unpublished observations).avian adenovirus was not detected. the depletion of lymphocytes and lymphocellular necrosis in the bursa of Fabricius and other lymphatic tissues lead to immunosuppression. such as bursa of Fabricius and spleen. the investigation of blood samples may be a good choice for the conformation of the diagnosis YPD in vivo. might result in enhanced viral replication. fungi and parasites. Comparable observations have been reported from other countries like Czech Republic. This might indicate that a (re)activation of the immune system has taken place due to the numerous of secondary infections commonly observed. However. coli may lead to an overcome of the barrier of the gut. In contrast PiCV DNA was not detected in blood samples of pigeons from lofts without clinical outbreaks of YPD. therefore. Furthermore. In accordance with recently published reports (TODD 2002. HATTERMANN 2002). and. first of all in lymphatic tissues. Belgium and Northern Ireland (TODD 2002). the amount of PiCV DNA in the organs was reduced. but only in 3 of 6 pigeons from lofts without YPD. coli. All of the pigeons with clinical signs of YPD were infected with PiCV. may overgrow the bacterial flora of the large intestine. PiCV DNA was also detected in the blood of most of the birds (48. this is in contrast to the situation in circovirus infected parrots (H GERLACH. especially yeast. Remarkably. indicate that the prevalence rate of PiCV in Germany is generally high. University of Leipzig. J Vet Diag Invest 1994. Avian Diseases 1994. J Vir Meth 2002. Evaluation of polymerase chain reaction and dot blot hybridisation tests in the diagnosis of pigeon circovirus infections. 89: 1 . 20. Escherichia coli infections in pigeons: Characteristics of the disease and its etiological agent. BARNES HJ. 1050 .16. TODD D. Adenovirus inclusion body hepatitis: A new pigeon disease? Proc Symp Avian Diseases.uni-leipzig. SHIVAPRASAD HL. Proc Assoc Avian Vet. Lake Worth 1994: 203 .379. VOGEL C. WOODS LW.14. 10. et al. 9. SHIVAPRASAD HL. Faculty of Veterinary Medicine. 6: 156 . Tauben.1053. BEARD CW. 104: 55 . DORRESTEIN G. An den Tierkliniken 17 D-04103 Leipzig.214. 3. RUPIPER DJ and EHRENBERG M.de (Footnotes) a mean value and standard deviation 165 . In: RITCHIE BW. Particles resembling circovirus in the bursa of Fabricius of pigeons.M. Lake Worth: Wingers Publishing. HATTERMANN K. A retrospective study of circovirus infection in pigeons: nine cases (1986-1993). 4. 38(3): 635 . Pigeon circovirus infection. Proc 9th Symp Avian Diseases. (eds): Diseases of Poultry.58. A method to diagnose pigeon circovirus infection in vivo.164. Necropsy examination. Munich 1994: 211 . HARRISON GJ and HARRISON LR (eds): Avian Medicine: Principles and Application. 5.6 CITATION INDEX 1. Department of Small Animal Medicine. WOODS LW. 7. Berlin: Deutscher Landwirtschaftsverlag 1992. Munich 1992: 7 . 6. DE HERDT P and VAN GINNEKEN C. LATIMER SL and RAKICH P. Germany Email: vschmidt@vogelklinik. Vet Microbio 2002. AUTHOR ADRESS V. 31. 10th edition. 1994.211. Inc. 355 . Introduction to pigeon medicine. In: CALNEK BW. 2. 8. Ames: Iowa State University Press 1997.M and VAN DER HAGE MH. Schmidt Clinic for Birds and Reptiles.641. D. triglycerides 116. Treviso. Italy SERUM BIOCHEMICAL AND ELECTROPHORETIC PATTERNS IN THE EURASIAN BUZZARD (BUTEO BUTEO) : REFERENCE VALUES.V. Electrophoresis can provide useful diagnostic information when used in conjunction with other laboratory tests.08g/l.37U/I.72mg/dl. total bilirubin 0. phosphorus 2. Electrophoretic patterns in serum or plasma protein are not available. PhD. D. total protein 3. ELECTROPHORESIS: pre-albumin 3. clinical evaluation. Besides. cholesterol 192.68 mg/dl. lipase 26.74g/l. �-globulin 4. Padua. Bedin3.. endoscopy and laboratory investigations (haemogram. D. PhD KEYWORDS Buteo buteo – Biochemistry – Haematology . faecal exams) were performed on each animal. biochemistry.8U/I.M.89g/l.15mg/dl.84 g/l. urea 12. due to age.78g/l..M.93U/I. L.1U/I.V.. alanine aminotransferase 40. D.03 mg/dl. creatinine 0.Department of Veterinary Clinical Science1. calcium 8. as patterns of alteration in protein electrophoretograms are commonly associated with clinical disease. �-globulin 5. bacteriology. Wildlife Rehabilitation Center2. but can be also physiological..4 mg/dl.67mg/dl.42mg/dl. In order to confirm health status. �-globulin 13.1 mg/ dl. aspartate aminotransferase 330. Our results can be used as a reference when evaluating electrophoretograms of diseased buzzards. lactate dehydrogenase 2008.Biol. alkaline phosphatase 89. so it is not always possible to perform electrophoresis as a routine test. F. magnesium 2.I.. We collected blood samples from 20 healthy Eurasian buzzards and carried out serum biochemical and electrophoretic tests.6U/I. glucose 337. WWF. D. D.4U/I.U. Ancona. sex. S. D.V. Romagnoli1. Wildlife Rehabilitation Center3.D. Bernardini1. The following mean values were obtained: BIOCHEMISTRY: albumin 14.Gelli1. 166 .M.. Ferrari1. V. S.M. especially in a Wildlife Rehabilitation Centres. amylase 616.Electrophoresis ABSTRACT Clinical and diagnostic usefulness of haematological and biochemical investigations in avian medicine is still a subject of debate.9U/I.6 g/l. gamma-glutamyl transferase 0. creatine kinase 1604. albumin 14.P. however some biochemical parameters of this species are still not available.2 mg/dl. this is probably due to the fact that wild bird medicine has little commercial interest.3U/I. M.V. changes in plasma proteins in birds are not exclusively related to diseases. Franceschini2.M. radiography.22mg/dl.06g/ l. 1 INTRODUCTION Many haematological and biochemical values in the Eurasian buzzard (Buteo buteo) have been published. uric acid 3.V. In capillary electrophoresis separation of protein fraction occurs in a borate buffer. The elecrophoretic profile is obtained by direct measurement of the proteins at 200 nm. Biochemical exams were performed by a 912 automatic analyser (Roche -Hitachi®). after clinical evaluation. endoscopy and laboratory investigations. was collected for pre-release routine investigations. Twenty of these animals were assessed as healthy. serum was preferred because fibrinogen in plasma can often obscure the electrophoretogram in the �-� region (THOMAS 2000). Serum was used to enable further biochemical and electrophoresis investigations to be performed.U. The serum samples for electrophoresis were diluted 1:25 with water prior to analysis to avoid the adhesion of proteins to the capillary walls. changes in serum proteins are detected early. both physiological and pathological patterns of electrophoretograms in the different species must be established. The birds were all adults of both sexes.I. in tubes without anticoagulant. W. Inc. The aim of this work was to characterise and document normal reference ranges and electrophoretograms patterns in the Eurasian buzzard. inflammatory processes of different aetiology). radiography. currently.1 Biochemistry Values of biochemical parameters found in the birds of this study are reported in Tables 1 and in Table 2 167 . Blood samples for biochemistry and electrophoresis were collected from the metatarsal vein.P. seroproteins were obtained using P/ACE MDQ capillary electrophoresis system (Beckman Coulter®. equipped with an automatic constant volume sample injection device and temperature control for the capillary. hinders the use of electrophoresis in many species of wild birds to provide diagnostic information. 3 RESULTS 3. This lack of data related to standard values. dehydration. even when serological tests fail to detect the infectious agent.W. In mammalian medicine electrophoretogram patterns provide information about increasing and decreasing levels of proteins with attention to single fraction and alterations in these fractions are pathognomonic of some specific diseases (e. To make a diagnostic tool useful in avian medicine. a useful diagnostic tool to evaluate body condition (starvation.g. Also. 1 ml from each bird.F and the University of Padua. Ca). In order to perform electrophoresis. to indicate a diagnosis and to monitor the animal during therapy and rehabilitation (BLANCO and HOFLE 2003).season and reproductive status (KURYE and GASPARSKA 1985). changes in total blood proteins and in some fractions like albumin and gamma globulins are. a monoclonal peak in the beta fraction may occur in patients with multiple myeloma or lymphoma). in the acute phase of some infectious mammal diseases. Systems control data collection and analyses were done with the software of the equipment. 2 MATERIAL AND METHODS From June to July 2004 the blood of 40 Eurasian buzzards waiting to be released after medical and surgical care performed during the previous winter at the Wildlife Centres of L. During the rehabilitation period the birds were fed with the same diet based on dead birds and rodents. in a narrow-bore uncoated fused silica capillary that is exposed to high voltages. In the avian patient. 15 mg/dl mg/dl mg/dl 12. total bilirubin (TB).89 g/l 5. alanine aminotransferase (ALT).65 g/l �-globulin 4.06 Units g/l . gamma-glutamyl transferase (GGT).22 mg/dl Mg 2.78 g/l Mean 3.72 8.3 89. urea.4 1604.67 Urea Ca P 2.5 3. glucose (Glu). creatinine (Crea).2 116.42 mg/dl Parameters UA 168 Albumin 14.93 U/I Parameters Alb TP Glu Mean 14.8 0. Crea 0.4 26. Values of serum protein parameters found in buzzards. lipase (LIP) and amylase (AMY). phosphorus (P) and magnesium (Mg) 3.84 337. calcium (Ca). cholesterol (CHOL).Table 1. creatine kinase (CK).03 192.08 g/l Parameters Prealbumin Mean 3. Table 2.1 Units g/l g/l mg/dl Albumin (Alb).1 0. triglycerides (TG).2 Electrophoresis Values of serum protein are reported in Table 3.9 U/I U/I U/I U/I U/I U/I mg/dl mg/dl mg/dl 40.6 2008. lactate dehydrogenase (LDH). �-globulin �-globulin 13. aspartate aminotransferase (AST). alkaline phosphatase (AP). Values of biochemical parameters found in buzzards. Values of biochemical parameters found in buzzards. total protein (TP).37 U/I ALT LDH CK GGT AP TB CHOL TG LIP AMY 616. AST 330. Table 3.68 Units mg/dl Uric acid (UA). As in other birds and mammals the globulins consist of a heterogenous group of proteins that have a weaker negative charge and migrate at a slower rate than albumin. Albumin is represented by a high peak shouldered by the � fraction. although the value of amylase can be considered within the ranges reported for the healthy avian patient. Both of these enzymes are sensitive to stress and physical exercise. which appeared to be within physiological ranges. A low to negligible pre-albumin fraction was present in serum of the healthy buzzards. but the variety of methods of analysis reported in literature does not allow us to evaluate the real baseline level of this enzyme. further studies are needed to evaluate if our results may have been influenced by the physical manipulation of the raptors. Values of triglycerides for Buteo buteo are not reported. Pre-albumin appears in the farthest area of migration. other parameters differed. all these birds were kept in large aviaries to allow them to fly. In B. and as this metabolite is also reported to increase after exercise and stressors factors (HOCHLEITHNER 1994). but within the ranges reported for avian patients (HOCHLEITHNER 1994). cholesterol. calcium. glucose. considering the levels of glucose and of other enzymes like alanine aminotransferase and aspartate aminotransferase. phosphorus and magnesium. Electrophoresis is gaining importance in avian medicine as a diagnostic tool and for monitoring the avian patient. we could hypothesise that the high levels of LDH and CK may be due to high daily exercise.4 DISCUSSION Although some of our biochemistry results were similar to previous reports such as: total proteins. migrating at a time of 7-7. 169 . Beta globulins values are slightly lower than the ones found in literature for other species of raptors. they migrate at a time of 4 to 6. so. urea.5 minutes.5 minutes. Although further studies are needed to improve knowledge. both on physiological and pathological electrophoretic patterns of protein fraction in serum and plasma of the European buzzard the values obtained in our study can be considered first reference values in this species. Gamma and beta globulins in this species of raptor appear in the electrophoretograms in a bridged peak and not as distinct fractions. Enzyme levels including creatine kinase and lactate dehydrogenase appeared to be very high despite the good physical condition of the buzzards. However. Amylase and lipase reference values for this species were not found in literature. Alkaline phosphatase appeared to be lower than in many other species of birds. buteo. which is an acute phase reactant of this fraction and can obscure the real value of this globulin. This is because we used serum instead of plasma to evaluate � globulins in healthy animals without fibrinogen. The blood was collected a few minutes after capturing the animals in the aviaries to avoid any increase in glucose level in serum due to stress and the level of this metabolite did not appear to be influenced by manipulation of the birds. Values of metabolites such as creatinine and uric acid appeared to be much lower than those reported by other authors for the European buzzard (HERNANDEZ 1990). J Avian Med Surg 12. AUTHOR ADRESS Donatella Gelli Department of Veterinary Clinical Science Viale dell’università 16. J Rapt Res 1990. 4 . ZINKL JG and JAIN NC (eds): Schalm’s Veterinary Hematology 5th ed. Tenerife 2003: 256 .261. CRAY C. QUESENBERRY K. 2.1109.502. Protein electrophoresis as a diagnostic and prognostic tool in raptor medicine. Lake Worth: Wingers Publishing 1994. Saunders Co.5 CITATION INDEX 1.212.10. 5.B. Plasma protein electrophoresis: an update. In: FELDMAN BF. Clinical Hematology and blood chemistry values for the common buzzard. HERNANDEZ M. HARRISON GJ and HARRISON LR (eds): Avian Medicine: Principles and Applications. protein electrophoresis and cholinesterase values of free-living nestling peregrine falcons in Spain. Reno 1997: 209 . New York: W. Baltimore: Lippincott Williams and Wilkins 2000. 35020 Legnaro. Padova. 223 . 7. Plasma protein electrophoresis as diagnostic and prognostic tool in raptors. 1107 . CRAY C. Italy Email: donatella. J Wild Dis 2001.it 170 . 6.245. Chicago 1991: 112 . 9. Biochemistries. Proc Euro Assoc Avian Vets.118. 899 . J Zoo Wild Med 2000. BLANCO JM and HOFLE U.177.gelli@unipd. Diagnostic use of protein electrophoresis in birds. LANZAROT MP. Protein electrophoresis. XIII. THOMAS JS. 31(4): 497 . Hematological. In: BONAGURA JD (ed): Kirk’s Current Veterinary Therapy. CRAY C. 37: 172 . TATUM LM. HOCHLEITHNER M. Proc Assoc Avian Vets.119. 8. 3. Applications of protein electrophoresis to avian diagnostics. 10. In: RITCHIE BW. Proc Assoc Avian Vet. 4. 2000.903. 14:113 . Plasma electrophoresis in psittacine birds. insectivorous and omnivorous birds tend to accumulate more iron in their liver and. stress stimulates iron absorption from the gut and may initiate haemosiderosis in birds. The disease is also frequently found in mynahs. Other dietary components (tannins.Transferrin ABSTRACT Although the exact pathogenesis of haemosiderosis in frugivorous birds has not been elucidated. 1 INTRODUCTION Haemosiderosis is a common finding in frugivorous birds kept in captivity. organic acids and minerals) significantly influence the bio-availability of the iron in the food and are at least as important. birds of paradise (Sturnidae). A higher intestinal iron uptake by mynah enterocytes compared to chicken enterocytes was demonstrated (METE 2003). Werquin DVM and K. and even some parrots (Psittaciformes). frugivorous. are more prone to the disease than carnivorous.Versele-Laga – R&D Department. Also stress reactions and defense mechanisms may induce iron accumulation in the liver: in contrast to mammals. quetzals (Caprimuligiformes). The most common group of birds to suffer from haemochromatosis are toucans.Iron storage disease . However. piscivorous and granivorous birds.Immune mechanisms .J.S. ascorbic acid. toucanettes and aracari (family Ramphastidae). This paper describes the different chemical forms of iron and their biological availability. This higher intestinal iron uptake could be due to a physiological adaptation to the low iron in the natural habitat of these birds. BUT SEVERAL DIETARY FACTORS AND BIRD-SPECIFIC DEFENSE MECHANISMS CONTRIBUTE TO THE DEVELOPMENT OF THE DISEASE G. Belgium HEPATIC HAEMOSIDEROSIS IN BIRDS : NOT ONLY TOTAL DIETARY IRON LEVEL. fibers. De Cock (Food engineer) KEYWORDS Birds .Haemosiderosis . In general. curassows (Struthioniformes). tanagers and starlings (Passeriformes). phytic acid. Deinze. there is a common consensus that dietary iron load should be restricted.L. current recommendations for total dietary iron are very empirical and disregard differences in bio-availability.D.J.Phytic acid . thus.Iron availability . where the duodenal epithelia are 171 .Tannin . programmed to take up all iron available. restriction of total dietary iron is limited. special premixes without iron containing additifs should be used. but shedded in the lumen of the gastrointestinal tract). Own research showed important variations in iron content of finished products. This paper inventarises factors that influence dietary iron load and describes how defense mechanisms may contribute to iron storage disease in birds. than to a variation in iron content. and levels of other minerals. indicating that other factors may also play an important role in the development of iron storage disease. the evaluation of dietary iron intake should not be limited to the absolute iron level of the diet. When formulating food for frugivorous birds. Further investigations of raw materials demonstrated that these variations where mainly due to the large variations of the iron content in 172 . Iron levels below 60 ppm are often difficult to realise without compromising dietary calcium supply. pH. the iron in the enterocytes is normally bound to ferritin and not transferred to the blood. chemical form of iron present in the diet and the balance of dietary components enhancing or inhibiting the absorption of dietary iron. As most available raw materials contain significant amounts of iron. Surveys of captive birds have produced arguments that there is a direct correlation between immunological or nutritional stress and haemosiderosis. Furthermore. They are often not the result of controlled feeding trials and disregard the variability in iron bio-availibility. which can lead to a > 10-fold variation in iron absorption. Therefore. the recommendations for dietary iron levels vary by author and seem to be set up rather empirically. it is generally accepted that genetic predisposition and dietary iron intake play an important role in the pathogenesis of iron storage disease. raw materials with low iron levels should be selected. It is well known that the variation in dietary iron absorption is due more to differences in the bioavailability. Manufacturers of feed for frugivorous birds should monitor iron content in finished products for each production batch. The iron content of vegetable materials is often very variable and depends on soil conditions as iron content. Raw materials used for the production of feeds for frugivorous birds should be analysed systematically on iron content. Also. it was observed that low iron diets do not protect birds from haemosiderosis. chemical form of iron.1 Dietary iron level Restriction of total dietary iron is the most obvious measure: Empirically based data suggest a maximum iron level of 100 ppm. However. as mineral sources are often rich in iron. These findings still do not explain why the mucosal block does not function (when body iron status is adequate. 2 DIETARY IRON LOAD AND HAEMOSIDEROSIS The amount of iron absorbed from the food is determined by iron status of the bird. but should also take the chemical form of dietary iron and factors influencing dietary iron absorption into consideration: 2. With the current knowledge. There is a common consensus that dietary iron load should be limited in birds prone to haemosiderosis. Concerning the labeling of iron content on the packaging of bird feed. it is important to realise that the European animal feed legislation normally only allows the labeling of the supplemented trace elements. an iron content < 5 mg / MJ ME has been proposed.mineral sources.low iron <300 alfalfa 300 oystershell 290 peas 263 corn gluten 200 brewers yeast 175 sorghum 170 flax 120 rape seed 90 wheat 40-75 corn 40-75 milocorn 40-75 paddy rice 50 fig 35 sunflower seed 30 walnut 25 peanut 20 honey 15 rose hip 5 juniper berry 5 raisin 5 173 .normal 2000-5000 calciumphosphate 800-1400 Meat 480 Spirulina 400 chalk . recommendations for iron levels should be expressed on energy basis in stead of on weight basis. The amount of iron mentioned under the food additives stands for the added iron: to obtain the total iron content. which is very variable for bird food. Some birdfeed companies however obtained derogations on this legislation and are allowed to mention the total iron content for feed destinated for frugivorous and insectivorous birds. The use of special chalk types with guaranteed low iron level proved to be essential in controlling the iron content. this amount must be increased with the iron content of the raw materials. As food consumption is determined by the energy content of the food.On energy basis. Table 1: Iron content in raw materials (mg/kg) chalk . The level of haem iron absorption is relatively unaffected by other dietary factors.3 Factors influencing iron absorption Several dietary factors significantly inhibit iron absorption. Haem and non-haem iron are absorbed differently. The majority of ferrous iron is found in haem iron and the majority of ferric iron is found in non-haem iron. typically less than 10% and often under 5%. Fe2+ (ferrous) and Fe3+ (ferric). supplementation of black tea leaves prevented iron absorption after feeding an iron-enriched diet (SEIBELS 2003). dietary iron inorganic organic Fe3+ Ferric Fe insoluble poorly absorbable Fe2+ Ferrous Fe soluble absorbable = iron porfyrines (haem) = better availability (different absorption mechanisms) = less influenced by dietary factors iron bio-availability Iron oxide < Iron(II)sulfate < iron chloride < iron citrate < iron amino acid complex < haem iron Fig 1: Bio-availability of different chemical compounds of iron Haem iron is present in the haemoglobin and myoglobin of animal products. Phytate. In wild-caught starlings. Also the chemical composition of non-haem iron is important: e. nuts and sorghum reduce the absorption of non-haem iron due to the formation of insoluble iron tannate complexes. bayberries. 2. Compared to haem iron it is relatively poorly absorbed.g. iron oxide is less absorbed than iron chloride or iron citrate. This form of iron is relatively available: about 30% of haem iron is absorbed from the diet. pulses and dried fruit.2 Sources of dietary iron Dietary iron is available in two valence states.2. Plant polyphenols such as tannins present in tea. strongly inhibits non-haem iron absorption by interacting 174 . Non-haem iron is found in materials of vegetable origin such as cereals. pulses (peas. present in the bran of wheat and other wholegrain cereals.5%) of iron uptake occurred at a ratio of 1:1 or less Fe to tannic acid (GLAHN 2002). whereas other factors enhance iron absorption. In vitro studies demonstrated that tannic acid is a very potent inhibitor of non-haem iron uptake. The presence of iron chelating agents in the food reduces the bio-availability of non-haem iron. as maximal inhibition (97. vegetables. beans and lentils) and nuts. The absorption of non-haem iron is much more influenced by the iron status of the animal and several components in the diet that can either inhibit or enhance its absorption. mango and lentils. copper. Also oxalic acid in green leafy vegetables may bind iron to form salts known as oxalates. manganese. rhubarb. rendering it less soluble. preventing iron absorption. iron overload lead. Also calcium and soil clay interfere with iron absorption. zinc ascorbate. ginger. and secondarily block iron through competitive inhibition. Table 2. Factors That Influence Iron Absorption Physical State (bioavailability) Inhibitors Competitors Facilitators haem > Fe2+ > Fe3+ phytates. soil clay. At a level of 1000ppm. soybeans. citrate. the effect was not located in the intestinal lumen but suggested competitive binding at a receptor of the mucosal cell. beans. Moreover. Sources of oxalates are most berries.with it. (a) 175 . iron deficiency Several other metals are taken up by the same absorption mechanism. plantains. strontium. Therefore. tannins. oxalates. tea. suggesting a common step in the transport of these 2 kinds of iron. A strong dose-effect relation between the amount of calcium and the reduction in nonhaem iron absorption has been observed in humans (HALLBERG 1991). calcium also inhibited the absorption of haem iron similarly. amino acids. Fig 2: Influence of different dietary components on iron absorption ratio. Especially zinc supplementation revealed to reduce iron absorption: in vitro studies demonstrated that zinc decreases iron uptake by 57 to 80% (GLAHN 2002). cobalt. bell peppers. phytic acid decreases iron absorption by about 60% (HALLBERG 1989). more iron deposition in the liver has been observed. Biochemical 176 . and to make sure that sufficient nutrients that inhibit iron absorption are present in the diet.Influence of ascorbic acid. Tannic acid is clearly the most potent inhibitor of iron uptake. Animal products should be avoided due to the highly available haem iron. Diet should also contain ingredients rich in tannins (sorghum. It has been suggested that these observations are the result of the iron withholding defense system. Fruits rich in ascorbic acid (citrus) should not be fed to birds prone to haemosiderosis. It may be interesting to develop a similar algorithm for bird nutrition. Another approach may be to assess available iron in bird diets by the use of in vitro methods. organic acids or a low dietary pH enhance iron absorption by reducing the ferric iron to the more readily absorbed ferrous form. HALLBERG AND HULTHEN (2000) developed an algorithm (fig. Due to the strong inhibitary effect of tannins and phytates. Adding tea to the drinking water may be helpful. The evaluation of the dietary iron load should not be limited to the iron content of the food ration. also the iron content of drinking water should be low. (b) Influence of tannic acid. 2) for predicting the effects of factors known to influence haem and non-haem iron absorption in humans. but formulated without tannins Ascorbic acid. it is important not to focus only on the iron content of the food. there are more and more indications that also other mechanisms play an important role in the pathogenesis of haemosiderosis. When evaluating dietary iron load. ascorbic acid) into consideration. black tea) and phytic acid (wheat bran). analysing only total iron content is not sufficient to assess available dietary iron in a reliable way. Particularly ascorbic acid is a strong promoter of iron absorption and has a counteracting effect on phytate and polyphenols. Derived from the algorithm of HALLBERG AND HULTHEN (2000). attempts should be made to take the factors that influence iron absorption (chemical form of iron. the evaluation of commercial feed for frugivorous food focused on the total iron content of these foods (SCHOEMAKER 2001). bacterial or fungal infections. (c) Influence of phytate phosphorus. but should also attempt to take all other important dietary factors into consideration. phytic acid. the amounts of these nutrients may be much more important than the iron content itself. stressful conditions. levels of tannic acid. The incidence of haemosiderosis is higher in birds which are kept in suboptimal. Of course. However. For practical feeding recommendations. (d) Influence of calcium. The tannic acid level is of more importance for the iron absorption than the total dietary iron: a sorghum based diet containing 350 mg tannic acid and 250 ppm iron gives less available iron than a low iron diet containing 60 ppm iron. Until now. 3 DEFENSE MECHANISMS AND HAEMOSIDEROSIS Besides dietary factors. Also in birds suffering from viral. 395. 3. Am J Clin Nutr 1991. Calcium: effect of different amounts on nonheme. blood iron is regulated by 2 proteins: lactoferrin and transferrin.384. transferrin is released in birds (HALLQUIST AND KLASING 1994. In birds. MAZUR-GONKOWSKA 2004). Serotransferrin. J Agric Food Chem 2002. et al. also in birds. 5. Both proteins have a high affinity for iron and are bacteriostatic in vitro for a number of bacteria. quarantine measures for new birds and preventive anti-parasitic treatments are also important. Biochem Mol Biol 1994. Com Biochem Phys B.144. SOUTH P and MILLER D. 49: 140 . BRUNE M and ROSSANDER L. Also transport stress should be limited as much as possible by the use of comfortable. et al. which binds the blood iron but also increases intestinal iron absorption and iron flux to the liver. KLAREN P. and lactoferrin (latest only in mammals) have a central role in ferrokinetics. KONCICKI A. tannic acid and ZnCl2: studies using an in vitro digestion/ Caco-2 cell model. HENDRIKS H. 177 . HALLQUIST N and KLASING K.evidence indicates that immunologic stress raises the level of iron-binding proteins that function in iron transport and that synthesis of these proteins is regulated by stress hormones. HALLBERG L and HULTHEN L. Iron-binding proteins such as transferrin. 71: 1147 . As in mammals. These iron-binding proteins also participate in the process of decreasing iron availability for the microorganisms. BRUNE M. 6.and heme-iron absorption in man. consistent feeding of an equilibrated diet at fixed feeding hours may further reduce stress. HALLBERG L. Am J Clin Nutr 2000. With these new insights. blood iron is regulated by transferrin only. XIE 2002. Am J Clin Nutr 1989. Bull Vet Inst Pulawy 2004. 48. 4. 19 . MAZUR-GONKOWKA B. During stress or infections. Iron metabolism in mynah birds 7. 2. ERLANDSSON M. Assessment of acute phase response in turkeys experimentally infected with Escherichia coli or haemorrhagic enteritis virus. 50(2): 390 . This mechanism may be more important than the dietary iron load in the pathogenesis of haemosiderosis. WORTLEY G. Stress can be reduced by optimal housing in spacious aviaries and avoiding overcrowding.119. KRASNOD and BSKA-DEPTA A. Regular. ovotransferrin and metallothionein levels during an immune response in chickens. HALLBERG L. Prediction of dietary iron absorption: an algorithm for calculating absorption and bioavailability of dietary iron. Optimal veterinary care with a disease control program. Iron absorption in man: ascorbic acid and dose-dependent inhibition by phytate. avoiding stress and infections are probably key factors in the prevention of haemosiderosis in birds. In mammals.23. ferritin.1160. withholding iron from potential pathogens has been described as a primitive host defense mechanism. large transport cages and appropriate feeding during transport. GLAHN R. 4 CITATION INDEX 1. Inhibition of iron uptake by phytic acid. 108 (3): 375 . 53: 112 . METE G. 126: 620 . Tijdschr Diergeneeskd 2001.623. 8. Effective use of tea to limit dietary iron available to starlings (Sturnus vulgaris). 9. LAMBERSKI N. HUFF W. SEIBELS B.werquin@verla. Kapellestraat 70. 32(6): 625 . and in particular the iron content. et al. et al.120. HUFF G. DVM R & D Department. Poult Sci. Avian Pathol 2003. Versele-Laga. GREGORY C. of commercial feeds for mynah birds. Belgium Email: guy. SCHOEMAKER N and BEYNEN A. AUTHORS ADRESS G. 10. Composition. 81(1): 112 .632. B9800 Deinze. 34 (3): 314 . WERQUIN. XIE H. Identification of ovotransferrin as an acute phase protein in chickens.(Gracula religiosa) resembles human hereditary haemochromatosis.be 178 . 2002. J Zoo Wild Med 2003.316. Histology of the pancreas showed lymphocytic infiltration and almost a total loss of tubuloacinar tissue. Faecal trypsin and amylase activities were decreased in comparison with reference values obtained from faeces of 24 racing pigeons. The Netherlands EXOCRINE PANCREATIC INSUFFICIENCY IN A RACING PIGEON (Columba livia domestica) O. 1 INTRODUCTION In domestic animals progressive loss of exocrine pancreatic acinar cell function leads to failure of digestion due to inadequate production of pancreatic enzymes. 179 .Section of Pet Avian. Utrecht University. The most common cause of such severe loss of exocrine tissue in the dog is pancreatic acinar atrophy (PAA).J.Birds. M.M. Exotic Animals and Wildlife Division.M. Dorrestein.Amylase. Amann. Westerhof. I. Aetiology. Visschers. Pancreatic insufficiency is caused less commonly by chronic pancreatitis and rarely pancreatic neoplasia (WILLIAMS 1989). Faecal examination revealed large amounts of fat and starch and lack of fatty acids. Chronic pancreatitis resulting in progressive destruction of pancreatic tissue is also a common cause of EPI in adult human beings. reproductive problems. food intake and faeces production decreased and the pigeon started gaining weight. After 21 days of hospitalisation the pigeon died following an acute onset of haemorrhagic nasal discharge. Schoemaker.T.Lipase. polyphagia and production of voluminous sand-coloured faeces.Coagulopathy ABSTRACT A six-year-old female racing pigeon (Columba livia domestica) was examined because of an 8-month-history of weight loss. diagnosis and treatment of exocrine pancreatic insufficiency and the pathophysiological relation between the haemorrhagic syndrome and the underlying disease will be discussed. Faculty of Veterinary Medicine. Lumeij KEYWORDS Pancreas. While supplementing the pigeon’s diet with pancreatic enzymes.J. N. G. J. The pigeon was hospitalised and fed with Harrison food (Juvenile FormulaTM. AST farma B. RITCHIE et al. 12. 4 CASE REPORT A 7-year-old female racing pigeon (Columba livia domestica) was presented to the avian and exotic animal division of the Faculty of Veterinary Medicine at Utrecht University with an 8 months history of weight loss (especially during the breeding season).In birds a decrease in acinar tissue or fibrosis can occur after a chronic inflammatory process and cause clinical symptoms suggestive of pancreatic exocrine insufficiency (GRAHAM 1992). also a case of pancreatic atrophy in a peregrine Falcon (Falco peregrinus) (SAMOUR and NALDO 2002) and in a blue and gold macaw (Ara ararauna) (QUESENBERRY and LIU 1986). Further tests were done to evaluate exocrine pancreatic function: faecal examination revealed the presence of starch and fat in absence of fatty acids. There was a marked leucopoenia 1. Fort Dodge. 1987) and the protein spectrum showed an increase of the gamma globulins. Harrison’s Bird Food. The Netherlands).3g/L (reference 1-3g/L.6. Oudewater. Brentwood. haematology and plasma biochemistry. therefore no data can be provided.3x109 /L. She was vaccinated once per year against Paramyxovirus I (Colombovac. 1997). the disorder studied here was previously described in a yellow naped Amazon parrot (Amazona ochrocephala) caused by an adenocarcinoma (RITCHEY et al. polyphagia and voluminous sand coloured faeces.. USA) mixed with pancreatic enzyme powder (Zymoral powder. but they were normal (493 and 3 U/L respectively) compared to the reference values reported in this study. Amylase and trypsin in faeces were also determined (0 and 2U/L respectively) and were below the reference values. therefore no data can be provided. Amylase and lipase where also measured.V. 2 MATERIALS AND METHODS This paper has been submitted for publication. 3 RESULTS This paper has been submitted for publication. 5 gram per 180 .1x109 /L (reference 13-22. LUMEIJ 1987) suggesting a chronic inflammatory process. On physical examination.5-3. Preliminary diagnostic work-up included faecal microscopical examination. the bird was alert but very lean weighing 372 g. LUMEIJ 1985) and subsequent decrease in the albumin/globulin ratio. EPI. The bird was hospitalised for diagnostic tests. inactivated adjuvant vaccine). TN.01 (reference 1. 0. Microscopically the lungs showed blood in the parabronchial lumen and capillaries and interstitial oedema. together with high amounts of fat and starch and lack of fatty acids. Some dogs with exocrine pancreatic insufficiency have high serum ALT activity because of disrupted small intestinal barriers and resultant hepatic inflammation. Severe haemorrhage was seen on the left leg from just above the tarsal joint to the left groin. but this would not exclude pancreatic disease in dogs where increases in amylase and lipase occur during acute pancreatitis or acute inflammatory crisis during chronic pancreatitis. Subnormal values indicate inadequate pancreatic exocrine secretion. Once again the appearance of the faeces went back to normal. The faeces returned to be voluminous and sand-coloured. was fed. The low faecal trypsin and amylase activities found is this pigeon. The final conclusion in the pathology report was pancreatitis. In dogs. Within 24 hours a decrease in bodyweight (10 %) and an increase in food intake (14 g more than the previous day) and faecal production were seen. coprophagia and sand-coloured voluminous faeces. 181 . a tentative diagnosis of exocrine pancreatic insufficiency is confirmed by measuring serum trypsin-like immunoreactivity (TLI) after a 12-hour fast. After one day of treatment an increase in the weight was observed. There was also a proliferation of the pancreatic ducts found. while the amount of faeces was smaller and was normal in colour. In dogs. After 21 days of hospitalisation the pigeon showed an acute onset of dyspnoea and haemorrhagic nasal discharge and it was placed in an oxygen cage. After a few days of therapy. Blood was found in the beak. On macroscopic examination a pale and skinny carcass was seen. results of history and physical examination differentiate primary small intestinal disease and malabsorption from exocrine pancreatic insufficiency. All of the symptoms the pigeon showed were similar to those seen in dogs and cats with exocrine pancreatic insufficiency: a chronic history of weight loss despite a vigorous appetite. trachea and lateral parts of both lungs. 5 DISCUSSION In this report. it was decided to challenge the pigeon and only Harrison baby bird food. qualitative faecal analysis for the presence of trypsin and amylase activities and undigested food particles can be used to arrive at a presumptive diagnosis. nephritis and bleeding due to trauma in combination with coagulation disorder. Langerhan’s islets from the exocrine part of the pancreas were not affected. Microscopy of the spleen showed proliferation of the white pulp. Microscopy of the pancreas revealed a large lymphocytic infiltration of the exocrine pancreas and an almost total loss of tubuloacinar tissue. without supplementation of pancreatic enzymes. Also. the occurrence of exocrine pancreatic insufficiency in a racing pigeon is described.100 gram Harrison food. indicate a problem with the production and/or secretion of pancreatic enzymes. Amylase and lipase values of the blood were normal. A few hours later it was found dead in its cage and was submitted for necropsy. Returning the pigeon on its original diet of Harrison and pancreatic enzymes once again resulted in an increase of bodyweight and a decrease in food intake and faecal production. 14: 401 . 97: 549 . J Am Vet Med Assoc 1986. which could extend to the lungs via the air sacs. also consistent with the presence of a chronic inflammation. 7. 3. 1996. Color atlas of avian histopathology. Necropsy indicated that the pigeon had died of haemorrhage.408. DUTTA SK. Blood chemistry reference values in racing pigeons (Columba livia domestica).Unfortunately. GRAHAM DL. The development of a coagulopathy in the course of exocrine pancreatic insufficiency can be explained by the inability of digesting and therefore absorbing food in the intestinal tract. 189: 1107 . This can result in a coagulopathy since vitamin K is essential in the pathways of secondary coagulation. microscopy of the spleen revealed comprehensive proliferation of the white pulp. Exocrine pancreatic insufficiency in a yellow-naped Amazon (Amazona ochrocephala) with pancreatic adenocarcinoma. and wild birds: a pathologist perspective.101. In EPI. exotic. and subsequently a vitamin K deficiency can develop as described in cats and humans (EDWARS et al. Avian Pathol 1985. Necropsy revealed inflammation and loss of exocrine pancreatic tissue. and REECE RL. which. RITCHEY JW. QUESENBERRY KE and LIU SK. 4. LUMEIJ JT. Diseases of the exocrine pancreas in pet. Ann Intern Med 1982. Probable vitamin K-deficient bleeding in two cats with malabsorption syndrome secondary to lymphocytic-plasmocytic enteritis. New Orleans 1992: 190 . This could be explained if the pigeon had a coagulopathy. TLI measuring techniques are not available for the racing pigeon and therefore the diagnosis could not be confirmed with this technique. The Vet Quarterly 1987. J Vet Intern Med 1987. Pancreatic atrophy in a blue and bold macaw. 1: 97 . 2.193. LUMEIJ JT. 6 CITATION INDEX 1. RANDALL C J. Furthermore. could cause excessive haemorrhage. minor trauma can lead to excessive bleeding and eventually death. 1987. 6. This step in lipid digestion is needed for adequate solubilisation and absorption of fat-soluble vitamins including vitamin K. DUTTA 1982). 9: 262 .1108. Vet Pathol 1997. based on the macroscopical finding of severe haemorrhage in the area of the left leg and the airways.268. 5.64. 34: 55 .57. EDWARDS DF. After developing such a coagulopathy. whereby indications for a chronic inflammatory process were found due to a leucopoenia and an increase in gamma. The diagnostic value of plasma proteins and non-protein nitrogen substances in birds. Proc Assoc Avian Vet. Unfortunately the pigeon died before all diagnostic testing could be concluded.52. 8. lipase deficiency prevents breakdown of dietary triglycerides into monoglycerides and fatty acids. Further research is therefore recommended in order to assess the possibilities of diagnosing EPI in vivo and the relationship between EPI and coagulopathies in avian species.globulins. 60 . 182 . consistent with the existence of exocrine pancreatic insufficiency due to chronic pancreatic inflammation and consistent with the results of blood analysis. Deficiencies of fat-soluble vitamins in treated patients with pancreatic insufficiency. in combination with trauma to the left leg. Yalelaan 8. Vet Rec 2002. DVM. AUTHORS ADDRESS Olga Amann.nl 183 . Email: o.9. Exocrine pancreatic disease. 3rd edition.125. Universiteit Utrecht. The Netherlands. Pancreatic atrophy in a peregrine falcon (Falco peregrinus).1554. WILLIAMS DA. 151: 124 .uu. 3584 CM Utrecht. Faculty of Veterinary Medicine. SAMOUR JH and NALDO J. Philadelphia: WB Saunders 1989. 10. In: ETTINGER SJ (ed): Textbook of veterinary internal medicine. Division of Avian and Exotic Animal Medicine Department of Clinical Sciences of Companion Animals.amann@vet. 1528 . The different techniques for endoscope guided multiple entry surgery in birds are described using sterilisation and castration as a model. but rarely used in avian medicine. One example is the visual assessment of the gonads to determine anatomical breeding functionality. despite the fact that imprinting parrots should always be avoided. Lierz. Special equipment of certain size and design (connectable to electro surgery) for use in avian medicine is commercially available. This is surprising. multiple entry surgical techniques can be applied in birds. Only endoscopy-guided tissue biopsies are routinely performed in avian medicine. Also imprinted aggressive parrots might be successfully treated with castration.Avian – Endoscopy-guided surgery . Hafez. Apart from surgical interventions through the working channel of an endoscope.M. endoscopic sexing has still advantages. Using these techniques it is also possible to sterilise and castrate birds of both genders. So far. In addition risks and problems using these techniques are discussed. endoscopy is a very valuable diagnostic tool within avian practice. In addition this procedure is important in female pet birds with chronic egg laying. In addition. In avian medicine endoscopic guided surgery is in its infancy and therefore seldom described. They are named as a potential genetic risk for the wild falcon populations (especially peregrines) in the case of mating with wild birds after escaping from captivity. As in human medicine. double and triple entry surgical techniques have been used. Germany ENDOSCOPY GUIDED MULTIPLE ENTRY SURGERY IN BIRDS M. endoscopy-guided surgery is a routine procedure and it is more and more used in small animal medicine.Minimally invasive surgery ABSTRACT Endoscopic guided surgery is a routine procedure in human medicine and commonly used in small animal practice. Prof Dr med vet KEYWORDS Castration . reducing their sexual behaviour.Sterilisation . 1 INTRODUCTION Avian endoscopy was developed in the 70´s for sexing monomorphic species. Despite establishment of DNA-methods for sexing.Institute for Poultry Diseases Free University of Berlin. This is very important as the Government of certain European countries are discussing this as a routine procedure in hybrid falcons. as the bird with its large air-sac system is ideal for 184 . Dr med vet and H. Castration and sterilisation of female pet birds to avoid a permanent egglaying. geese) (CONVERSE and KENELLY 1994).such procedures and insufflations pumps are not necessary. Beside the quality improvement of meat (castration of bulls and boars). Examples are: Production of falcon hybrids and species conservation projects (A female bird of a rare species is stimulated by a partner of a common species but inseminated with sperm of the rare species. But comparable to small animal medicine also bird owners require such procedures for their animals for the following reasons: Castration of hand reared parrots of both sexes to avoid a sexual related aggression against the owner. Well known and regular used is a single-entry technique. other surgeries will be indicated such as tumour or granuloma resection. finches. Recently endoscopyguided surgery in birds has been developed (HERNANDEZ-DIVERS 2004. Hybrids species (birds of prey. . This has already been performed by the authors successfully with an air sac tumour in a hybrid falcon. Therefore the instrument cannot be manipulated independently of the endoscope. just the castration of male chickens (caponization) is routinely performed to modify the meat quality.Castration and sterilisation to control free ranging populations (feral pigeons. Using a double entry technique an additional cannula with trocar (as channel for an instrument) is placed. A single instrument is directed through a working channel into the visual field of the endoscope. With the use of these techniques. The approaches to the avian coelom are chosen according to the procedure performed. therefore one rare male can be used for several females). Last but not least. Basically different techniques can be distinguished using endoscope guided surgery. as it represents a lower impact to the bird compared to a laparotomy. In avian medicine. (Comm.: Therefore hand.Sterilisation of male birds to use them as stimulation partners for females (used for artificial insemination with sperm of other males) (SAMOUR and MARHAM 1987). . the modification of behaviour (castration of male dogs). Recently a governmental request has been made for castration and/or sterilisation of birds. waterfowl) are often produced but there 185 . in triple entry techniques two cannulae are placed in addition to the centrally placed endoscope. the control of populations (sterilisation of feral cats) or the adaptation to the owner for an easier keeping (castration of female dogs) are reasons for such interventions. These should be used as practical model for introducing endosurgery into avian medicine. . The present paper describes the use of endosurgery techniques in sterilisation and castration of birds as an example.).rearing should always be avoided. 2 STERILISATION AND CASTRATION Sterilisation and castration are routine procedures in veterinary medicine. enabling the surgeon to work with two different instruments. LIERZ 2004). Sterilisation and castration of birds are regular requested procedures and can ideally be performed using endosurgery. 5mm (according to the instrument size) cannulae with trocars. Using this approach nearly all organs can be evaluated. Because of the advantageous angle to the gonad and the larger surgical field the following approaches are recommended. Within the wild they might be able to reproduce with the nominate species introducing foreign genetic material. The development of 2 and 3mm human paediatric laparoscopy equipment (Karl Storz Inc. Larger surgical intervention can be avoided and the risks. It is minimally invasive and the gonads located at the cranial pole of the kidney are easy to reach. SURGICAL TECHNIQUE 3. plastic or graphite. Equipment The main problem using endosurgery is the maintenance of haemostasis. The material of such a unit is stainless steel. Therefore a radiosurgery unit for the connection to the endosurgical instruments is vital. scissors. Placing the birds identical the pelvic limb is pulled cranial. One cannula is inserted caudal to the endoscope through the obliquus abdominis externus muscle just caudal to the mid-point of the pubis. After inserting. The instruments are introduced into the surgical field using 2. 3. Surgical Procedure Preparation for sterilisation or castration by endosurgery (single or double entry technique) is similar to a routine endoscopic examination pulling the left pelvic limb caudal (LIERZ 2004). the trocar is removed holding the cannula in place. Firstly. 3.7mm 30° telescope with a working channel. In addition to the basic endoscopic equipment (2. as described for castration of chickens (COLLIGNON 1949). In addition a bipolar forceps and a monopolar sling are essential. Advanced surgeons might extend their equipment to palpation probes. This double entry technique can be extended by introducing a second cannula cranial to the endoscope into the last or second last 186 . laser units can be used but they are almost not routinely available in veterinary practice.2. As prohibition of producing such hybrids is not always possible or useful to castrate or sterilise such hybrids is in discussion. flexor cruris medialis the endoscope is introduced caudal to the last rib. which can also be used for the other above described techniques. flexible biopsy and grasping forceps as well as scissors) a variety of instruments are available and necessary. the approach to introduce the endoscope is chosen caudal to the last rib. needle holders and knot tiers. The additional cannula using the double entry technique is placed cranioventral to the endoscope between the last two ribs. placing the second trocar ventrocaudal to the endoscope.is a possibility that they may escape.5mm or 3. Within the USA hybrid falcons have to be imprinted or sterilised prior their use in falconry. Castration and sterilisation of birds by endosurgery seem to be ideal. As imprinting leads to unwanted behaviour modification in the birds the sterilisation becomes important (JONES and REDIG 2003).1. grasping forceps from which parts should be monopolar and therefore connectable to the radiosurgery unit. Also. Using the triple entry technique the same approaches can be used. The approach is from the left side (female birds) or from both sides (male birds). Ventral to the m. can be minimised.) has forced the endosurgery in birds. This includes dissection forceps. In juvenile birds the deferent duct can be grasped and removed by pulling using a grasping forceps through the endoscope working channel. In adult birds.intercostals space. According to our experience the ovary stay juvenile in some of those birds. Such females are developing uneventful (incl.2. In some cases it was noticed that the oviduct fills with egg albumin during sexual activity and this might lead to problems. The endoscopist controls the endoscope and the instrument by using the single or double entry technique. Therefore the triple entry technique is only possible in birds larger than 400g. Using a grasping forceps the testicle is elevated from the underlying tissue. Now the oviduct is cut between both obliterations using a monopolar scissor. Therefore female birds are sterilised rather than castrated. As an alternative the deferent duct can also be obliterated by electrosurgery. The infundibulum can be grasped and the oviduct can be removed from underlying tissue by gently pulling. Using a second entry port a scissor is introduced cutting the deferent duct.2. Sterilisation of male birds Sterilisation of male birds is described using a laparotomy (BIRKHEAD and PELLAT 1989). By using a multiple entry technique it is very important to keep the approaches of the instrument ports and the endoscope as far apart from each other as possible. The radiosurgery unit is activated by a foot. The same is performed at the most caudal part of the oviduct which can be reached. As an alternative the oviduct can be obliterated by a monopolar or bipolar grasping forceps. In addition it is recommended to remove at least 1 cm of the duct to avoid a re-establishment of functionality if both ends reunifies.2. Therefore the complete removal of the oviduct is recommended. 3. In juvenile birds a grasping forceps can be introduced through the endoscopes working channel (single entry technique). Performing a triple entry technique the endoscope is placed on a sand bag or handled by an assistant while the endoscopist controls both instruments. By this way it is possible to remove nearly the entire oviduct without greater bleedings. It is of advantage to obliterate the deferent duct in two places and to cut it in between. not expecting any disadvantages as observed in female birds. The oviduct is fixed by a grasping forceps and obliterated as far cranial as possible using a mono or bipolar grasping forceps through a second entry port. Sterilisation and castration of female birds The ovary is a very fragile organ and its endoscopic removal represents a high risk of lethal bleeding. as it was shown in cockatiels (PYE et al.pedal. For sterilisation and in most cases castration the single or double entry techniques are sufficient. In addition the sexual behaviour of such males was unchanged (JONES and REDIG 2003). ovary) and are hormonal active. 187 . 2001). In addition the total removal of all hormone-producing tissue is very difficult. as described in human medicine.1. Using this method in Japanese quail it was demonstrated that the testicles developing uneventful and that the testosterone blood levels increasing with sexual maturity similar to un-sterilised males. Ovulation was not noted in such birds. 3. In adult birds endoscope guided sterilisation is only possible using a double or triple entry technique. A second obliteration is performed slightly behind the first one. The endosurgery technique is similar to those of female birds. leading also to an elevation of the deferent duct. the oviduct can only be removed using a double or triple entry technique. Personal communication. BIRKHEAD TR and PELLATT JE: Vasectomy in small passerine birds.3. Using coagulation techniques it is vital to avoid any damage to underlying tissues especially the kidneys and ureters. Proc Euro Assoc Avian Vet. 22: 112 . 6. LIERZ M: Endoskopie.94. COLLIGNON P: Das Kapaunisieren – Die Kastration der Junghähne und anderen Geflügels. Wildl Soc Bull 1994. 2004. 120: 115. 2. RISKS The risk of uncontrolled lethal bleeding is always present during endosurgery. 117 . 15: 90 . The mesorchium can be cut using a monopolar coagulation scissor. Especially in adult birds.A. Afterwards the testicle can be removed. kidney. In adult birds. 185 . In: ROEMER R GEFLÜGELZUCHT – BÜCHEREI.. Vet Rec 1987. PYE G. CONVERSE K.194.2. obliterating the vessels when closing the sling. Heft 7.W. JONES R. 8.): Leitsymptome bei Papageien und Sittichen. The testicle is elevated from the kidney using a grasping forceps through the working channel of the endoscope (double entry technique) or an extra entry port (triple entry technique).3. the triple entry is of advantage as it allows a better view to the surgical field.123. blood vessels) or attached to other organs (depending on the species) leading to further difficulties. Stuttgart: Eugen-Ulmer Verlag. 4. the blood supply of the testicle is increased making an additional obliteration of the blood vessels by monopolar or bipolar coagulation forceps necessary. In: Pees M (ed. Stuttgart 2004.117 HERNANDEZ-DIVERS S. the structures of the genital tract are sometimes difficult to distinguish from other tissues (adrenal gland. Vet Rec 1989. The risk of lethal bleedings is very high in male birds during sexual activity and therefore castration must be avoided during that time. BENNETT RA. ureter. This requires more experience and training. Enke Verlag. Therefore sterilisation and castration of juvenile birds is always preferable as the blood supply of the genital tract is low. On the other hand. J Avian Med Surg 2001. ISBN: 3-8304-1023-9. 1949. 188 .H. Tenerife 2003. and KENELLY JJ: Evaluation of Canada goose sterilisation for population control. In juvenile birds the testicle can be grasped and removed by a radiosurgical sling. 4. 7. 3. 5. CITATION INDEX 1. PLUNSKE R and DAVIDSON J: Endoscopic salpingohysterectomy of juvenile cockatiels (Nymphicus hollandicus). SAMOUR J. and MARKHAM JA: Vasectomy in budgerigars (Melopsittacus undulatus). 125: 646. and REDIG PT: Endoscopy guided vasectomy in the immature japanese quail (Coturnix coturnix japonica). Castration of male birds The castration of male birds is only possible using a double or triple entry technique. de 189 .michael@vetmed. Free University of Berlin. Königsweg 63.AUTHORS ADRESS Michael Lierz Institute for Poultry Diseases. Germany Email: lierz. 14163 Berlin.fu-berlin. 8mt from the floor. The femoral head is conserved to improve mechanical functions of the hip. Signs developed after they had fallen from the beam with their leg caught. O. the anaesthetic and surgical techniques used and the good results obtained using this surgical technique. Gauthier DVM. The two birds were adult and weighed 1.Cormorant ABSTRACT (Phalacrocorax carbo) were two captive cormorants presented with a femoral head dislocation. France TWO CASES OF FEMORAL HEAD LUXATIONS IN CORMORANTS (PHALACROCORAX CARBO) TREATED WITH A MODIFIED MEIJ-HAZEWINKEL-NAP TECHNIQUE E. The injury occurred during a show in a zoo.Coxofemoral luxation . National Veterinary School of Nantes. PhD KEYWORDS Bird . with one leg restrained with a rope. A first treatment with capsulorraphy failed. They lived in a zoo and participated every day in a bird show.Surgery . the femoral head was in a good position. D. The other bird recovered over several months. Risi DVM. 1. the birds were kept on a beam. 190 . It was decided to try a modified Meij-Hazewinkel-Nap technique to treat these dislocations. Between each performance. The technique consists of reducing the dislocation and fixing the greater trochanter to the hip using a synthetic prosthesis. We will present the clinical and radiological aspects of the dislocation. Ordonneau DVM. One bird died a few weeks post-surgery from probable non-related causes. 1 INTRODUCTION Two cormorants (Phalacrocorax carbo) were seen due to a lack of activity and reluctance to move for 4 days.The Wildlife Center.8 and 2kg respectively. Leg paresis. A severe unilateral non-weight-bearing lameness was present in the two cormorants. By forcing them.Positive nociception test: strong reaction of the bird but difficult movement of the leg. Femoral head or neck fracture. the two birds were in sterno-abdominal recumbency and refused to move.Asymmetric position of the two legs in extension. the injured leg appeared internally rotated and longer than the healthy one. before falling down again. Anaesthesia 191 .2 MATERIAL AND METHODS Clinical examination At the clinical examination. . . Differential diagnoses The differential diagnosis included: Ventral luxation of the femoral head. 10mg/kg IV) via a catheter in the axillary vein. . Other examinations Radiographs in ventrodorsal and lateral recumbency confirmed the ventral coxofemoral luxation of the right leg and the left one respectively. Ventral luxation of the femoral head associated with a proximal fracture of the femur. 0.Negative proprioception test.Pain at coxofemoral mobilization . the following signs were present: . . Ventral luxation of the femoral head associated with nervous damaged.Lack of tonicity of the damaged leg. . Ventral luxation of the femoral head associated with acetabular fracture. followed by ketamine (Chlorketam.Slight amyotrophy of the damaged leg. The first cormorant was anaesthetized using an injection of midazolam (Hypnovel. 3 TREATMENT A surgical treatment was made 7 days after the accident.5 mg/kg IM).Abnormal latero-lateral movements of the damaged leg when the bird was held over the floor. The birds were in a good condition and no other abnormality was found. The clinical exam of the musculoskeletal system revealed some abnormalities on the right leg of one bird and on the left one of the second bird. . No other lesions were visible. Proximal fracture of the femur. An open reduction technique with capsular suture and non-absorbable sutures between the periosteum of the greater trochanter and the acetabular rim was performed. they could stand up with difficulty for only a few seconds on one leg. In the two cases. between the greater trochanter and the ilium by perforating them. This technique had to be modified. Anaesthesia was induced with diazepam (Valium. Radiography revealed a new ventral luxation. 4mg/kg sid IM for 7 days) were given. 1992. Time and intensity of this re-education were daily increased. surgery was performed. Bandages were removed when going back to the zoo. made with a non-absorbable material. because of the hip anatomy of the cormorant and the biped posture. The good reduction of the luxation was confirmed by a radiograph. caudally and dorsally to the hip articulation. A curved skin incision was made over the greater trochanter. Ketofen. associated with an amyotrophy and a moderate muscular fibrosis. abduction and traction of the leg. Analgesia was provided by nalbuphine (Nubain. the leg was held abnormally. A moderate lameness was 192 . 0. The articular cavity was gently cleaned by removing all the coagulated blood and necrotic tissue. The ventral dislocation was then reduced by internal rotation. was present for the 10 first days after bandage removing. the same bandage. 10mg/kg bid for 10 days) and NSAID (ketoprofen. Marbocyl. Necrotic gluteal muscles borders were debrided before suturing these muscles with an absorbable material (Safil). 4 RESULTS The two birds were hospitalized for 3 weeks while bandaged. A Kirshner pin was introduced through the acetabular protuberance and then through the greater trochanter in order to make tunnels in each of these bones. In order not to damage the healthy cranial soft tissue. between the already damaged muscles. The gluteal muscles were torn. Five sutures with a non-absorbable suture material (Flexiden) were made through the gluteal muscle tendons on their trochanteric attachments and the acetabular rim periosteum. The prosthetic ligament (a 1/0 non-absorbable woven multifilament suture) was then passed trough these tunnels and tied in a figure-of-eight pattern. in a bent position for performed. A modified Meij-HazewinkelNap technique was attempted (MEIJ et al. Regular radiographs have been taken to make sure of the good position of the articulation. The day after surgery.5mg/kg) and ketamine (10mg/kg) by intravenous injection then isoflurane.5mg/kg IM). Then. Cage confinement and limited activity were required. This technique consists of stabilizing the hip by passing a prosthetic ligament. 1999). The leg was then maintained in a bent position using a bandage under the body. reduce amyotrophy and joint ankylosis. Skin was then closed with U points. The damaged leg was plucked and disinfected with iodine povidone (Vetedine). Moreover. the birds were allowed to swim in their pool giving further leg movements. The luxation of the second cormorant was treated using the same technique.was then maintained with isoflurane after intubation. The capsular lesions were too severe to allow a capsulorrhaphy. The day after. A non-weightbearing lameness. Antibiotics (marbofloxacin. MEHEUST et al. The caudal border of the acetabulum and the caudal protuberance (present in cormorants) were gently exposed by removing the muscles around. it was decided to use a caudal approach to the hip. A re-educational program was established: the leg was handled daily with rotating movements of the knee and the hip in order to make muscles work. 0. The bird was placed in a lateral recumbency on the healthy side with the wings pulled dorsally. one of the cormorants was found dead without clinical signs. because of the bird’s bipedal posture. The ventral position of the femoral head is here simply explained by the trauma history: a fall from a beam with a caught leg. MEHEUST et al. 1994. MEIJ et al. BENNETT 1997). FOSSUM 2002. luxations are not a common problem and are less frequently reported than fractures (MARTIN and RITCHIE 1994. due to abnormal or compensatory weight-bearing (MACCOY 1989). 1994. hip luxation may rapidly cause problems in the nonluxated leg. no necropsy was made only an incision was made on the operative site to confirm the good position of the coxofemoral articulation and the absence of infectious or abnormal inflammatory reaction around the prosthesis. 1999).g. transacetabular pin (through the great trochanter into the head of the femur across the acetabulum) or extra-articular stabilisation with a prosthetic ligament between the femoral neck and the ilium (Meij-Hazewinkel-Nap technique) (BRINCKER et al. 1994). The different techniques of stabilization include: capsule reconstruction (capsulorrhaphy). 1994. HOLSWORTH and DECAMP 2003). Soft tissue damage always includes round ligament and articular capsule rupture (BRINCKER et al. HOLSWORTH and DECAMP 2003). hip luxations are frequently due to trauma (BRINCKER et al. In birds. In mammals. A prosthetic capsule technique or a toggle pin procedure is preferred (FOSSUM 2002). they are more generally craniodorsal to the acetabulum (MARTIN and RITCHIE 1994. the luxation must be treated to prevent periarticular fibrosis formation leading to joint ankylosis (BENNETT 1997). over a 13 week period after surgery. HOLSWORTH and DECAMP 2003. suture of the dorsal joint capsule alone is inadequate to maintain reduction. 1994. the luxations were ventral and associated with gluteal muscle damage. Gluteal muscle damage is seen in severe cases (BRINCKER et al. In our 2 cases. 193 . Unfortunately. excision arthroplasty of the femoral head and neck. by closed or open reduction (BRINCKER et al. translocation of the greater trochanter.then observed over the following weeks and locomotory function improved gradually with swimming and re-education sessions. Craniodorsal luxations are the most common type (70-80% of patients) (BRINCKER et al. MACCOY 1989) and occur with tendons afflictions or more frequently with trauma by traction and rotation of the leg (HEIDENREICH 1997.2% of patients) and frequently associated with acetabular fractures (BRINCKER et al. HOLSWORTH and DECAMP 2003).5-3. FOSSUM 2002. Luxations in birds should be reduced as early as possible (less than 3 days) (BENNETT 1997). bumblefoot. For ventral luxations. Elbow and shoulder luxations are more common than coxofemoral dislocation (HEIDENREICH 1997). pseudarthrosis. 1994. The second bird gradually returned to full activity. muscle contracture and chronic deviation of the limb (MACCOY 1989). 1994). FOSSUM 2002). In birds. BENNETT 1997. e. prosthetic round ligament. MACCOY 1989). Moreover. hip luxations must be treated quickly. prosthetic capsule (modified Knowles toggle technique). 1992. FOSSUM 2002. As in mammals.Ventral luxations are rare (1. Seven weeks after surgery. 5 DISCUSSION In pet mammals. with a slight lameness but a normal range of motion. They may be considered as rare and serious luxations. 1999). But. The caudal approach of the articulation was made here to avoid other muscle damages and it is reported that. due to long time between the surgery and the death (7 weeks). In spite of the recommendation to place the prosthetic ligament cranial to the articulation (between the greater trochanter and the ilium. Meij-Hazewinkel-Nap technique (MEIJ et al. Few other methods are reported in birds. in birds. because of the anatomy of the leg. 1992. The modified Meij-Hazewinkel-Nap technique used in our cases seems to give good results in birds. a death due to surgery seems unlikely. MEHEUST et al. This long immobilization time could have been responsible for the muscle contracture seen after removal. the caudal approach is preferred to avoid potential injury to the cranial coxal artery and to provide better exposure of the femoral head and neck (MACCOY 1989). Without post-mortem examination. Transacetabular pinning is not recommended because of the risk of penetrating the kidney by inserting the pin too deep (leading to kidney lesions and haemorrhages) (BENNETT 1997). a real Ehmer bandage was not possible. 1994. In mammals. we preferred a caudal position (between the greater trochanter and the acetabular caudal protuberance) to avoid instability by pushing on the leg in the biped position. we decided not to use this technique to prevent gluteal muscles perforation by the femur. The birds kept this bandage for 3 weeks to prevent recurrence of the luxation. but no cases have been reported. In our case. 1994. The forces applied to the hip are almost vertical in birds because of this biped position and the ligament seemed to have a better function in a caudal position. an Ehmer bandage is made to assist hip reduction (4-7 days postoperatively) (BRINCKER et al. The birds weighed between 554g and 1355g). very few techniques are described for hip luxation treatment.In birds. even if this possibility can not be excluded. FOSSUM 2002). No damage to the sciatic nerve occurred with this approach in our two cases. we cannot be sure that the cause of death was unrelated to surgery. A manual reduction and application of a hip splint has been reported in a hyacinth macaw but the bird did not tolerate the splint and frequent recurrences occurred (MACCOY 1989). Owing to the larger weight of the two cormorants and their poor and injured gluteal muscles masses. The results were good several years after surgery. and the absence of gross lesions around the prosthesis. BENNETT 1997). this technique requires a second anaesthetic to remove the pin after a few days. The bandages should have been removed earlier and physiotherapy begun sooner to have better and quicker results. MACCOY describes an excision arthroplasty technique used in 3 birds (a hyacinth macaw. HOLSWORTH and DECAMP 2003). Limited activity is required for an additional 3 weeks (FOSSUM 2002. to maintain the femoral head into the acetabular cavity. Reduction and stabilization with supporting structures (non absorbable sutures). Moreover. The animal is then gradually returned to full activity over a 2-3 weeks period (BRINCKER et al. FOSSUM 2002). An immobilization was made in order to hold the leg in a bending position under the body. in the author’s knowledge. 194 . placed from the trochanter to the dorsolateral iliac crest and to the cranial rim of the acetabulum are proposed (MARTIN and RITCHIE 1994. in order to prevent degenerative joint disease development (BENNETT 1997). a Moluccan cockatoo and an African grey parrot) with a caudolateral approach to the articulation (MACCOY 1989). The recovery time to full locomotion may have been prolonged by this immobilization. Lake Worth: Wingers Publishing 1994. 5. CLUBB SL. Physiotherapy in water is required to improve hip mechanical functions.326. 2. Prat Med Chir Anim Comp 1999. NAP RC.766. DVM Centre de Soins de la Faune Sauvage. Oxford: Blackwell Science 1997. (34): 611 . 194(1): 95 . Editions du Point Vétérinaire 1994. Manuel d’orthopédie et de traitement des fractures des petits animaux. 7. BRINCKER et al. In: SLATTER I (ed): Textbook of small animal surgery 3th ed. J Small Anim Pract 1992. In: RITCHIE BW.621. 560. Orthopedic surgery. 1400. HARRISON GJ and HARRISON LR (eds): Avian medicine. HOLSWORTH IG and DECAMP CE. Stabilisation des luxations coxofémorales traumatiques par la technique de Meij-Hazewinkel-Nap modifiée: étude rétrospective de 63 cas. 7 CITATION INDEX 1. LEGEAR F. St Louis: Mosby 2002. MACCOY DM. Philadelphia: WB Saunders Co 1997.2008. 284.1169. AUTHORS ADDRESS Emmanuel Risi. 733 . Philadelphia: WB Saunders 2003. Orthopedic surgical techniques. QUESENBERRY K (eds): Avian medicine and surgery.6 CONCLUSION A modified Meij-Hazewinkel-Nap technique can be used in cormorants for coxofemoral luxation treatment. MEHEUST P. MARTIN H and RITCHIE BW. FOSSUM TW. 4. 9.risi@wanadoo. Ecole Nationale Vétérinaire de Nantes. (2): 2002 . Maisons-Alfort. BENNETT RA. 1137 .97. A bandage must be applied after surgery and removed before 3 weeks. Small Animal Surgery 2nd ed. (33): 320 . BOURGERON A. J Am Vet Med Ass 1989. MEIJ BP. Birds of prey. principles and application. HEIDEN REICH M. France Email: Emmanuel. medicine and management. DORRESTEIN GM. 3. Route de Gachet. 44300 Nantes. HAZELWINKEL AW. Results of extra-articular stabilisation following open reduction of coxofemoral luxation in dogs and cats. 6. Coxofemoral luxation.fr 195 . 8. In: ALTMAN RB. School of Veterinary Medicine. bony changes and alignment of the joint. Villaverde DVM1. R. Muñoz Guzon DVM PhD2. functional angles. Lugo. 196 . University Complutense of Madrid. Freire DVM1. Spain MANAGEMENT OF THE FEMOROTIBIAL LUXATION BY COAPTATION SPLINTING. The articular surfaces had a minimal damage. Gonzalez DVM1. M. The functional angles were similar in external skeletal fixators (ESF) before and after the surgery. There were not significant histological differences among the different groups when evaluating periarticular fibrosis tissue and bone reaction. Rodriguez-Quiros DVM PhD1 KEYWORDS Avian – Birds – Pigeons – Stifle – Luxation – Orthopaedics ABSTRACT Repair of a stifle luxation requires stabilisation to minimize the formation of periarticular fibrosis. INTRAMEDULLARY PINS. J. Animals were weight bearing two or three days later with splinting and IM pins techniques. Madrid Department of Veterinary Clinical Sciences2. Lopez Peña DVM PhD2. All the techniques performed represented a viable option to luxation repair.Department of Surgery and Internal Medicine1. Animals were assigned to six groups. J. The initial goal consisted of providing lack of femorotibial stability. EXTERNAL SKELETAL FIXATOR AND TENSION BANDS: A PILOT STUDY IN DOMESTIC PIGEONS (Columba livia domestica) S. M. Domestic pigeons were selected. F. Decreased anaesthetic and surgical duration was seen with single band tension and splinting. Benito de la Vibora DVM1. We evaluated anaesthetic and surgical duration. Open surgical managements may increase morbidity. The alignment was restored except in the control group. The purpose of this study is to show comparatively different surgical techniques for luxation repair of the femorotibial joint. School of Veterinary Medicine of Lugo. University of Santiago de Compostela. mobility and normal use of the limb. The formation of scar tissue to stabilise the joint requires 3 to 6 weeks. inhibiting reduction of the luxation and predisposing to joint ankylosis (BENNETT 1998). The femorotibiofibular joint is luxated with concomitant damage of the ligaments (Bennett 1998) and rarely meniscal damage is diagnosed (HARCOURT-BROWN 2000). Butorphanol (1 mg/kg IM) and meloxicam (0. Within a period of as little as 3 days. CLIPSHAM 1991). The total fluid volume was 5% weight equally divided between the two injections. with a mean body weight of 290 g were selected for the pilot study. Dislocation of the femorotibial joint with damage to the collateral ligaments generally results from a developmental abnormality (BENNETT 1998. The birds were induced with isoflurane by mask and they were maintained on a surgical plane of anaesthesia using an oxygen fresh gas flow through a modified Jackson Rees non-rebreathing circuit adjusted to 0. as early as possible to minimize the formation of periarticular fibrosis (BENNET 1998.2 mg/kg IM) were administered preoperatively. MARTIN and RITCHIE 1994). but has been mainly diagnosed in psittacines and raptors (BOWLES and ZANTOP 2002. craniomedially. Stifle luxation is uncommon in birds (ROSENTHAL et al. This luxation is diagnosed definitely radiographically (BOWLES and ZANTOP 2002).9 L/minute and isoflurane inspired concentration adjusted to 2%. positive withdrawal sign and also medial or lateral instability on examination. The pigeons were housed indoors and were allowed to acclimatise for at least two weeks before the start of the experimental procedures which were performed after obtaining authorization from our Hospital Institutional Animal Care Committee. caudolaterally and caudomedially. Findings include hyperextension of the joint. 1992). Luxation of the femorotibial joint occurs craniolaterally. The purpose of this pilot study was to show comparatively five different surgical techniques for luxation repair of the femorotibial joint in birds. and the ability to partially reduce the tibiotarsus into its normal anatomic position suggested a luxation instead of a fracture. 197 . Anaesthetic monitoring included electrocardiogram (ECG). BOWLES and ZANTOP 2002. lack of crepitus on palpation. Repair of a stifle luxation requires stabilisation of the joint by reduction and immobilisation. HARCOURT-BROWN 2000).1 INTRODUCTION Stifle luxation studies and case reports in different species of exotic mammals and reptilian have been reported. respiratory rate. relative arterial oxygen (SpO2) and pulse rate by use of a pulse oximeter. Absence of palpable fracture. 2 MATERIAL AND METHODS Eighteen mature domestic pigeons (Columba livia domestica). Hydration with lactated Ringer’s solution injected subcutaneously was made for each pigeon before induction and at the completion of each surgery. significant fibrosis occurs. spontaneous orthopaedic disease (HARCOURT-BROWN 2000) and following traumatic episodes (BOWLES and ZANTOP 2002). There are scanty reports in the literature showing comparatively surgical techniques for stifle luxation repair in avian patients (MACCOY 1988). but the closure was made with a 4-0 synthetic absorbable suture material of polyglycolic acid to suture the joint capsule incision. and then collateral ligaments were also sectioned to allow a mediolateral stifle luxation. Lübeck. VTP®) for the connecting bars. The leg was wrapping with a cohesive flexible material (i. the subcutaneous tissue and the skin in a simple continuous pattern. A linear incision was made cranial to the lateral collateral ligament. A splint was applied from femur to metatarsal bones including the digits I and III to keep the joint immobile and to maintain the pigeon’s leg in a proper position. Germany). It was applied with the joint placed in a functional angle of 60º. the initial goal of the experimental design consisted of providing lack of femorotibial stability. Group 5 consisted of a screw and tension band with a single strand of nylon suture. A pin of 1. After aseptic preparation and draping. The external portion of each pin was bent 90º form its insertion portion and then they were joined with a veterinary thermoplastic material (i. Group 4 consisted of a transarticular type I ESF.e. All of the surgical and radiographic procedures were performed under general anaesthesia. One 1. A second hole was drilled through the fibular crest with a 3-0 nylon suture needle. Before the skin incision was accomplished. consisting of three birds per group. VTP®) to maintain the stifle joint placed in a functional angle of 60º. On this way withdrawal sign was positive on palpation of the stifle.5 mm Ø bone screw was placed into the femoral condyle. The wires were bent using a veterinary thermoplastic casting material (i. lateral collateral ligament was identified. The splint with a normal angle was attached by wrapping with an elastic tape (i. Vetrap®) around the leg. Group 2 consisted of coaption splinting.e.0 mm Ø) were fixed into the lateral aspect of the femur (2) and tibiotarsus (3). by the management performed to stabilise the luxation: • Group 1 served as control group since the capsule was incised without repairing the joint. Following dislocation of the femorotibial joint. Through the hole a 7 mm long.end tidal carbon dioxide partial pressure (ETCO2) and rectal temperature (Julian DRÄGER anaesthesia machine and monitoring. lateral collateral and femorofibular ligaments were sectioned to achieve a medial patellar dislocation.e. from lateral condyle of the femur to the fibular head. The luxation was reduced with a tension band of a single strand of 3-0 nylon suture that extended from the screw in the lateral condyle of femur to the predrilled hole into the cnemial crest. Vetrap®) in a Robert Jones bandage to protect the surgical repair. Group 3 consisted of two intramedullary pins (IM). The surgical procedure was initiated with a lateral approach to the stifle.0 mm Ø Kirschner wire was placed into the medullary cavity of the tibiotarsal bone and the second one was seated into the medullary cavity of the femur. The joint capsule was incised.e. pigeons were randomly assigned to six groups. 1. Five K-wires (1. • • • • 198 .2 mm was used to drill a hole through the condyles of the femur. 2 mg/kg orally q24h. Four µm sections were stained with haematoxylin and eosin for histopathology. In these groups some cases of articular ankylosis and calcifications were observed. Analgesia was provided with buthorphanol (1. Enrofloxacin (10 mg/kg IM q12h) was administered prophylactically for a minimum of 5 days following surgery. 3 RESULTS All the eighteen procedures were performed safely and easily. in order to value some aspects such as periarticular fibrosis tissue and bone reaction and to compare definitely the different surgical techniques. The luxation was reduced similar to group 5 and a second strand of nylon suture was fixed from lateral condyle of femur to the space between the fibula and tibiotarsus. Decreased anaesthetic and surgical duration was seen in management with single band tension and splinting compared with other techniques. decalcified. The joint was observed radiographically without evidence of potential complications or bony changes in the groups. The stifle was taken out for histophatological study at 6 weeks post surgery. 199 . No inflammatory changes were presented. Animals recovered from anaesthesia. group 6 consisted of a cortical bone screw and double tension band. the bony changes and alignment of the joint radiographically. Animals were weight bearing on the limb two or three days later with splinting and IM pins techniques. The postoperative care was the same for all the groups. before and after the surgery. Histological changes were observed only in groups treated with IM pins and ESF. The functional angles of the flexing limb were similar in ESF technique before and after the surgery. morbidity and normal use of the limb putting weight. the functional angles of the limb flexing and extending the knee. There were not significant histological differences among the different groups when evaluating some aspects such as periarticular fibrosis tissue and bone reaction. for three or four days). The stifle alignment was restored except in the control group.0 mg/kg IM q6-8h) or meloxicam (0. Postoperative radiographs were taken to confirm that the reduction was made and remained intact.• Finally. Healing of the joint was assessed before removal of the orthopaedic material. ventral to the fibular head. Samples were fixed in 10% buffered formalin. The return to normal function of the limb putting weight after surgery suggests that the articular surfaces of the stifle had a minimal damage. bearing the foot and ambulating. processed by routine histological methods and embedded in paraffin. This study evaluated the anaesthetic and surgical duration of the different managements. is the poor anatomic reduction to maintain stabilisation of the joint. 200 . to ambulate and to balance on the other limb (MACCOY 1988). Reduction and stabilisation of the femorotibial joint by surgical fixation is considered the best option because of the patient’s small size. ROSENTHAL et al. ESF with connecting bars with polymethylmethacrylate or stainless steel may be used in avian patients (BENNETT 1998. and age (BOWLES and ZANTOP 2002. HARCOURT-BROWN 2000). The use of methacrylates in birds instead of stainless steel connecting bars of ESF. casts and splinting. Evidence of the return to normal function of birds after stabilisation is by its ability to use the limb to bear weight. The repair of damaged ligaments in large birds using suture material may be attempted (HARCOURT-BROWN 2000). SATERFIELD and O’ROURKE 1981).4 DISCUSSION It is very important to reduce the luxation as early as possible to minimize the formation of fibrosis. ROSENTHAL et al. Techniques describing IM pins joined with acrylic to rigidly stabilise the knee in its normal anatomic position have been reported (BOWLES and ZANTOP 2002). The procedure used to repair the dislocation depends on the size of the patient and the nature of the injury (BENNETT 1998). 1992). 1992). HARCOURT-BROWN 2000. 1992). ESF do not allow limb growth in paediatric patients (BOWLES and ZANTOP 2002). 1992). post-luxation bumblefoot. allows readjustment until the proper anatomic position is achieved (ROSENTHAL et al. compromising its function and range of motion. The significant disadvantage of these techniques is that the pins could interfere with normal stifle joint function (MACCOY 1988). Open surgical management and internal or external fixation techniques have been described (ROSENTHAL et al. Stifle exploration in avian species may be difficult because of the vulnerability of the neurovascular bundle to damage (BOWLES and ZANTOP 2002). The formation of sufficient scar tissue to stabilise the joint requires 3 to 6 weeks. with increased anaesthetic and surgical duration and increased soft tissue trauma. Although these techniques save the limb. ROSENTHAL et al. Cortical bone screws with band tensions have a significant disadvantage because once the screw is placed. degenerative changes and abnormal weight-bearing (BENNETT 1998. The femorotibial joint can be stabilised by using close or open techniques. sequelaes include increased morbidity and prolonged recovery. The main disadvantage of close managements such as coaptation by bandages. Complications of ESF include pathological fracture of the femur or tibiotarsus. which inhibits the reduction of luxation predisposing to joint ankylosis since it is assumed that joint luxation carries a poor prognosis (BENNETT 1998). it is impossible to readjust the angle in which the stifle become fixed (ROSENTHAL et al. REDIG 2001. Arthrotomy has been used to repair femorotibial joint injuries in birds (BOWLES and ZANTOP 2002). which is the interval before the pins are removed (BENNET 1998). Procedures with band tension have been reported with cerclage wire and other different suture and different splinting techniques (HOLZ 1992). 1992. 1992). weight. Recovery may be adversely affected by excessive weight of the fixator and decreased patient compliance (BOWLES and ZANTOP 2002. ROSENTHAL K. Luxation of the stifle joint in a Major Mitchell cockatoo. for their help. CD-Rom. 6(4): 235 . Effective methods for management of avian fractures and other orthopaedic problems. 6. 5 ACKNOWLEDGEMENTS The authors gratefully acknowledge Pilar Llorens. International Wildlife Rehabilitation Council (IWRC).38. and coaptation splinting. Inc. REDIG PT. 2000.204. BW. and especially to Isabel Garcia. (1997). 7. MARTIN HD and RITCHIE BW.15. 201 . 16(1): 34 . 1137 .167. 1998. California. CLIPSHAM R. J Am Anim Hosp 1981. chief of the radiological service. HYLLYER E and MATHIESSEN D. HARRISON.637. J Assoc Avian Vet 1992. radiology technicians of Imaging Diagnostic Section of the Veterinary Teaching Hospital. Without their competence and persistence in the technical and nursing care of the patients. Birds of Prey: Anatomy. We want to express our gratitude to all the students workers. HARCOURT-BROWN NH. plating.42. 3. Concord. Zoological Education Network. in the Veterinary Teaching Hospital. 9. 4.238. Correction of pediatric leg disorders.A bridge to the 21st Century (Proceedings of the International Wildlife Rehabilitation Council Conference. Carmen Osorno and Sonia Pavon. A novel surgical technique for luxation repair of the femorotibial joint in a monk parakeet (Myiopsitta monachus). 6 CITATION INDEX BENNETT RA. 17: 635 . 10. Proc Eastern States Vet Med Assoc 1988: 166 . 8. Orthopedic surgical techniques. Florida. Lake Worth: 1994. Management of joint luxations in birds. Stifle luxation repair in a moluccan cockatoo and a barn owl. Proc Assoc Avian Vet. Munich 2001: 26 . MACCOY DM. pinning.. 11 . 130(2): 34. 1. Wingers Publishing Inc. BOWLES HL and ZANTOP DW. Exotic and Wild Animal Surgery Group. In: REYNOLDS MD (ed): From Science to Reality . GJ and HARRISON LD (eds): Avian Medicine: Principles and Application. Proc Eur Assoc Avian Vet. External skeletal fixation in avian orthopedics using a modified through-and-through Kirschner-Ehmer splint technique (the Boston technique). the success rate of orthopaedic procedures we enjoy would not be realised. 2. SATTERFIELD WC and O’ROURKE KI. HOLZ P.1169. We conclude that stifle luxation repair in small birds might be accomplished with surgical techniques whichever stabilise the joint and permit a good clinical evolution. Chicago 1991: 200 . radiology and clinical conditions of the pelvic limb. In: RITCHIE. Management of coxofemoral luxations by femoral head ostectomy and long bone fractures by K-E splint. Vet Rec 1992.The results of this preliminary study indicate that all of these techniques performed represent a viable option to femorotibial luxation repair in avian species. J Avian Med Surg 2002. Suisun City. Lake Worth. 5. especially to Ruben Mota and Alicia of the Avian. California. Asistencia Veterinaria s. C.I. Sevilleja de la Jara. Villaverde ESVA. Centro de Estudios de Rapaces Ibéricas.l. Spain Email:
[email protected] 202 .E. 45671 Toledo.AUTHORSS ADRESS S.. Psittacine . Baton Rouge. The 2 beak deviation presentations have been classified as scissor beak. SPEER 1995 and 2003). but at the very least. In supporting the corrective device the bird’s natural beak dynamics allow for greater force and quicker manipulation of the beak back into its normal position without the consequences often involved with previously published techniques. N. T. Louisiana. DVM. MARTIN and RITCHIE 1994). MS. continued beak trimming is required due to the malocclusion and 203 . In severe cases the bird’s ability to eat may be compromised. although other psittacine species may be affected by either of these conditions. SPEER 1995). MS. Scissor beak most often affects macaw species while mandibular prognathism is diagnosed commonly in cockatoos. Dip ECAMS.Beak deviation .Mandibular prognathism Scissor beak ABSTRACT Psittacine species have 2 common beak deviation presentations that present during their growth phase. Mitchell DVM.The Louisiana State University. Diaz . O.Figaroa. United States of America USE OF A DENTAL COMPOSITE TO CORRECT BEAK DEVIATION IN PSITTACINE SPECIES T. G. M. where the lower beak is longer than the upper beak causing the upper beak to grow into the lower beak (CLIPSHAM 1989 and 1992. School of Veterinary Medicine Department Veterinary Clinical Sciences. where the rostrum maxillare (upper beak) grows to the side of the rostrum mandibulare (lower beak) and mandibular prognathism. There have been a number of corrective devices promoted to correct these beak deviation conditions in parrot species (CLIPSHAM 1992. Diplomate ABVP (avian). Stevens. A. PhD KEYWORDS Birds . DVM. Tully. Zachariah DVM. Jr. The advantage of this technique is that it uses a cold rapid curing human dental composite and the bird’s natural beak dynamics to support the corrective device.Dental composite . DVM. A. INTRODUCTION Scissor beak and mandibular prognathism are common beak deviation presentations in captive raised psittacine species (CLIPSHAM 1992. Once there is an understanding of the primary goal: function not perfection and acceptance of this goal then the process of correction can begin. The bird is placed in an avian restraint board (Miami Vice. The dental acrylic is layered on the tip of the upper beak for mandibular prognathism and the lower beak for birds presenting with scissors beak. Once there is a substantial amount of dental composite applied to the beak the Dremel® tool with the sandstone bur is used to modify the mass into the corrective shape. A disadvantage with this new technique is similar to other corrective hardware placements. For most cases weekly revisits over the first 6 weeks of treatment are required to assess the patient’s condition and response to the corrective device. Modifications are made to the composite by applying new layers of material over the cured appliance. USA) and held in a standing position. Reforming the composite must take place after the material cures. The beak speculum and tongue depressors are left in place as the acrylic cures. The composite will fall off the beak but appears to remain longer on the beak than the ultraviolet cured acrylic products. Illinois. Another disadvantage of previously published beak correction techniques is the use of general anaesthesia to apply the acrylic or hardware. Boca Raton. Birds with deviated beaks also are difficult to sell or bring a reduced price because of their undesirable appearance. the acrylic foundation breaking off from the beak with pieces of the beak attached and iatrogenic beak trauma from the forces applied leaving permanent beak scarring. Robert Bosch Tool Corp. Dental Products. Illinois. An avian beak speculum (Lafeber Products. Veterinary Specialty Products. 3M ESPE AG. The composite can be cut off the beak in pieces and if the material does not fall off can be pulled off by using a slight twisting motion. MATERIALS AND METHODS The patient and owner are initially assessed on compatibility for this long term (3 to 6 month) intensive beak correcting process. cool curing and the use of the bird’s natural beak anatomy for support of the composite mass. By using the bird’s natural beak anatomy to support the composite mass the chance of iatrogenic beak damage is reduced and gives more support for correction as pressure is place to move the beak into normal position. Cornell.. Mount Prospect. USA) is used to open the beak and tongue depressors are used to manipulate l the dental acrylic. This may also include forming a trough in the composite to manipulate the beak toward centre for scissor beak cases. the patient may have to be hand or tube fed during the treatment period. 204 . The dental acrylic (Protemp™ 3 Garant™. usually 5 to 7 minutes. The other advantages include rapid.continued growth of the keratin surface of the beak. D-82229 Seefeld – Germany) is then placed on a beak that has been scored using a Dremel® tool (Dremel®. the pins pulling through the beak and/or frontal bone. Florida. The problems that are often encountered with previously published techniques is the dental acrylic having difficulty adhering to the beak surface. Previously published techniques have used ultraviolet cured dental acrylic extensions either placed directly over the beak surface or onto a wire/plate foundation to push the beak back into place (CLIPSHAM 1989) and pins/wires placed through the beak and frontal bone using rubber bands to pull the beak back into place (MARTIN and RITCHIE 1994). USA) fitted with a sandstone bur. With this new technique the bird is placed in an avian restraint board and the corrective composite is applied while the bird is awake. Proc Assoc Avian Vet.176. Pittsburgh 2003: 45 . A few cockatoos with mandibular prognathism have also been treated using the dental composite. Proc Euro Assoc Avian Vet. If an owner is willing to hand feed a bird. HARRISON LR (eds): Avian Medicine: Principles and Application. 5. possibly throughout the corrective period.1169. composite falling off prior to visit and owner compliance problems with follow up visits. 1: 22 . SPEER BL. CLIPSHAM R. We have found that this technique offers a quick correction in the scissor beak cases while reducing the number of iatrogenic damage associated with previously published beak corrective techniques. California: Raintree 1995. Proc Ass Avian Vet. Semin Avian Exotic Pet Med 1992.49. wires and dental bar foundations. DISCUSSION This technique is not for every beak deviation case but it can provide many advantages to previously published techniques. Lake Worth: Wingers 1994. MARTIN H and RITCHIE BW. Orthopedic surgical techniques. 205 . Tenerife 2003: 3. Surgical beak restoration and correction. SPEER BL. the majority being macaw species presenting with scissor beak. 1165 . Fort Bragg. There should be minimal if any pain associated with this procedure and the bird is awake during the application of the composite appliance. THOMSEN JB (eds): The Large Macaws. The success we have noted with this dental composite technique gives clients hope that their animals will not end up with permanent scarring associated with pins. In: RITCHIE BW. 4. Non-infectious disease of pediatric psittacines. Trans-sinus pinning technique for the correction of chronic mandibular prognathism in psittacines. 6. A technique for correction of chronic mandibular prognathism. HARRISON GJ. perfection of deviation correction not obtained. By using the natural physics and anatomy of the beak the dental composite correction gives a rapid. SPEER BL. Problems that have been encountered with this technique include.RESULTS This beak correction technique has been tried on approximately 10 birds. hand feeding the patient. make routine appointments and call if there is a change in position of the device then they are good candidates for having their bird treated. Seattle 1989: 164 . 323 .324. 2. Non-infectious diseases. CITATION INDEX 1.33. 3. SPEER BL. secure adjustment to the malformed beak. CLIPSHAM R. In: ABRAMSON J. Diplomate ABVP (avian). Dip ECAMS Louisiana State University.lsu. Skip Bertman Dr. MS. DVM. Tully. Department Veterinary Clinical Sciences.. Baton Rouge. Jr.edu 206 . School of Veterinary Medicine.AUTHORS ADDRESS Thomas N.. United States of America Email: ttully@vetmed. Louisiana 70803. MCMILLIAN 1994). DVM. United States of America Loro Parque-Clinica Veterinaria Puerto de la Cruz. The purpose of this study is to determine if the use of brief isoflurane anaesthesia for the collection of radiographs during a contrast study slows the progression of a contrast media when compared to the use of manual restraint alone. acetylpromazine meleate and triflupromazine hydrochloride were studied using fluoroscopy. HOWLETT 2000. Dipl. Pentobarbital sodium. KRAUTWALD-JUNGHANNS 1996. and to distinguish the gastrointestinal tract from other celomic structures. Tenerife. promazine hydrochloride. methoxyflurane.Avian and Exotic Animal Clinic of Indianapolis Indianapolis. Crosta. Several small animal radiology texts mentioning a transit time decrease with the use of anaesthetics in dogs refer to a single study in 1973 using 2 dogs. stating that anaesthesia may slow gastrointestinal transit time. fentanyl. ABVP-Avian. halothane. 1 INTRODUCTION Nearly all avian medicine and surgery textbooks advice against the use of any anaesthesia in the collection of radiographs during a contrast study.M. It is assumed authors of radiology sections of these books base the recommendations against anaesthesia on the current understanding of the effects of anaesthesia on the gastrointestinal transit time of dogs and cats. Not one of four leading avian textbooks offers a specific reference to support this claim (SMITH BJ and SMITH SA 1997. L. Most drugs decreased gastrointestinal motility. Most textbooks and references describing the use of contrast advise against the use of anaesthetics during a contrast study. DVM KEYWORDS Isoflurane . Lennox. meperidine. droperidol. Spain THE EFFECTS OF ISOFLURANE EFFECTS OF ISOFLURANE ANAESTHESIA ON GASTROINTESTINAL TRANSIT TIME A.Anaesthesia ABSTRACT Oral contrast media is frequently used in psittacine radiology to evaluate the structure and motility of the gastrointestinal tract.Gastrointestinal transit time . with 207 . especially in birds with gastrointestinal disease. and struggling. fear and struggling. significantly decreased transit time. while ketamine and ketamine/acepromazine. The content of food items also affects motility. Psittacines resent being secured to a radiograph cassette. then avian practitioners should no longer be advised to avoid anaesthesia for that reason alone. The authors are unaware of a similar study utilizing psittacines. and regurgitation of contrast material is a risk. as the procedure produces at least some degree of excitement. The purpose of this study is to compare sets of radiographs using contrast media with isoflurane and with manual restraint alone. A rough estimation of gastrointestinal transit time might also be without radiography by measuring the time it takes for coloured markers. 208 . If the use of isoflurane does not significantly slow gastrointestinal transit time when compared to manual restraint. Another study in cats demonstrated that IV diazepam also decreased gastrointestinal transit time (HOGAN and ARONSON 1988.” Of interest is the comment that even “minor psychic disturbances” of the subjects caused at least some delay in gastrointestinal transit time (ZONTINE 1973). Certain antibiotics have been documented to reduce gastrointestinal transit time in poultry. A study using cats in 1988 investigated the effects of a number of injectable anaesthetics on gastrointestinal transit time. 1997). For example. while the presence of food (GI distention) and increased length of fasting increases motility. Evaluation of the gastric transit time alone in psittacines is likely best accomplished in a non-anesthetized. it is unlikely a study on the effect of isoflurane alone on gastrointestinal transit exists. As inhalant anaesthetics are rarely used as a single anaesthetic agent in dogs and cats.halothane and methoxyfluorane producing “profound depression. non-restrained and acclimated bird utilizing fluoroscopy. STEYN et al. Studies in poultry indicate that crop motility is significantly inhibited by excitement. The use of manual restraint alone to collect radiographs involved some degree of difficulty. foods of high fat and protein content cause a slowing of gastrointestinal time. producing poor-quality radiographs. It should also be considered that some excitement and struggling occurs during administration of isoflurane as well. especially crop or upper GI stasis. ventriculus and duodenum are then coordinated in sequence. Ketamine/diazepam combinations produced the least effect on transit time. Manually restraining a psittacine for radiographs is very likely to decrease gastric motility. with a decrease normally seen at night (DUKE 1986). personal communication). Non-anesthetized birds may struggle at the moment the radiograph is collected. Non-anesthetized birds cannot be intubated. to appear in the stool (SILVERMAN S. Numerous factors are known to influence gastrointestinal motility and GI transit time in mammals and birds. fear. or in a similar situation with a perching patient using standard radiographs collected via a lateral beam. Diurnal rhythms affect motility. Isoflurane as a sole agent is the most commonly used anaesthetic in avian medicine. Contractions of the proventriculus. even with the use of sophisticated avian restraint devices. such as vegetable dye. and the amount remaining in the crop. g. Each individual bird’s radiographs with and without isoflurane were compared. or simply taped to the cassette. Species Sex Age Comments App. radiographs were again collected using the same amount of barium and at the same time intervals. Barium sulphate was administered into the crop at a dosage of 25ml/kg. and radiographs were collected without anaesthesia at set intervals using either a commercial manual restraint device. Rather. No attempt was made to compare one bird to another bird. g. Radiographs were subjectively examined for a significant difference in the level of barium in the lower gastrointestinal tract. practitioners should no longer avoid anaesthesia during these procedures for that reason alone. isoflurane anaesthesia. 209 . proventriculus. with isoflurane administered via facemask for the duration of the brief radiographic procedure. galerita Sulphur crested cockatoo C. galerita Cockatiel Nymphicus hollandicus Cockatiel Nymphicus hollandicus Cockatiel Nymphicus hollandicus Cockatiel Nymphicus hollandicus Cockatiel Nymphicus hollandicus M F M F M F M F F M M F F Unknown Unknown Unknown Unknown 9y 11y Unknown Unknown Unknown Unknown Unknown Unknown Unknown Same Same Same Same Same Same Same Same Same from mixed viary with PDD Same Same Same 3 RESULTS No significant differences were detected in the progression of barium between radiographs collected with isoflurane and manual restraint alone. and ventriculus at the different time intervals. sex. Summary of birds used to compare gastrointestinal time with manual restraint vs. as it does isoflurane anaesthesia. decisions on whether or not to use anaesthesia should be based on other pertinent factors. Table 1. species and physical health were used in the study (table 1). After several days. Therefore.2 MATERIALS AND METHODS Fifteen birds of varying age. such as patient condition and perceived risk. It is assumed that stress and excitement during manual restraint produces the same effect on gastrointestinal transit time. healthy 1 Red fronted macaw Ara rubrogenys M 3y on PE 2 Red fronted macaw Ara rubrogenys F 2y Same 3 4 5 6 7 8 9 10 11 12 13 14 15 Blue and gold macaw Ara ararauna Blue and gold macaw Ara ararauna Green winged macaw Ara chloroptera Green winged macaw Ara chloroptera Little corella Cocatua sanguinea Little corella Cocatua sanguinea Sulphur crested cockatoo C. Lennox Avian and Exotic Animal Clinic of Indianapolis. Alimentary canal: anatomy. In: STURKIE PD (ed): Avian Physiology. Dipl. (ed): Avian Medicine. Effect of sedation on transit time of feline gastrointestinal contrast studies. Vet Radiol Ultrasound 1988. HARRISON GJ and HARRISON LR (eds): Avian Medicine: Principles and Application. 5.79. regulation of feeding and motility. SMITH BJ and SMITH SA. PhD. 268 . In: SAMOUR J. 28 . STEYN PF. DORRENSTEIN G and QUESENBERRY K (eds): Avian Medicine and Surgery. Personal communication: SILVERMAN S. United States of America Email:
[email protected] 210 . 8. In: ALTMAN R. New York. Effect of chemical restraint drugs on the passage of barium sulfate through the stomach and duodenum of dogs. DVM. 29(2): 85 . 630 . 2. ZONTINE WJ. Indianapolis.473. KRAUTWALD-JUNGHANNS ME. NY: Springer-Verlag 1986. 4. Clinical and diagnostic procedures. Avian radiology. American Academy of Veterinary Radiology. Vet Radiol 1997. London: Mosby Co 2000. HOWLETT JC.4 CITATION INDEX 1. 9. In: RITCHIE BW. 7. 3. Imaging techniques. Baltimore: Williams and Wilkins 1996. The effect of intravenous diazepam on solid phase gastric emptying in normal cats. CLUB S.663.88. 162: 878: 878 . HOGAN PM and ARONSON E. 170 . MCMILLIAN MC. In: ROSSKOPF WJ and WOERPEL RW (eds): Diseases of cage and aviary birds. 6. J Am Vet Med Assoc 1973. Lake Worth: Wingers Publishing1994.884. 28(6): 469 . Philadelphia: WB Saunders 1997. DUKE GE. TWEDT D and TOOMBS W. Radiology. AUTHOR ADDRESS M.326.288.199. 246 . Strong correlations (p < 0. Each bird was anaesthetized once via face mask. Respiratory rates were controlled to achieve five levels of PETCO2 from 20 to 60 mm Hg. Université de Montréal. These results suggest that the Nellcor NPB-75® capnograph could be a useful non-invasive monitoring tool for the evaluation of CO2 dynamic in birds over 400 g. Rondenay. DMV. (anaesthesiology). CA. as well as between PETCO2 and arterial pH. were used for this study.Monitoring ABSTRACT We compared the value of end-expired partial pressure of carbon dioxide (PETCO2) measured with a handheld capnograph (NPB-75®. assisted ventilation is recommended to prevent hypercapnia in bird anaesthetized with isoflurane (FORBES 1999). G. Analysis of blood gas in arterial 211 .Birds of prey . Lair. heart rate and body temperature were monitored during the whole procedure. Faculté de médecine vétérinaire. such as limited cardiovascular depression. low hepatotoxicity. Rés. DMV.Avian .Capnometry . Canada MONITORING OF END-EXPIRED PARTIAL PRESSURES OF CARBON DIOXIDE IN ANAESTHETIZED BIRDS OF PREY M. Concomitantly measured values of PaCO2 were determined for each level of PETCO2 obtained. intubated and manually ventilated. Arterial blood pressure. DES. Cert. Québec. IPSAV.0001) were observed between PETCO2 and PaCO2. DVSc. Birds are especially sensitive to the effects of this respiratory depression since both inspiration and expiration are active processes. DVM. Diplomate ACZM KEYWORDS Anaesthesia . Despite numerous qualities. MSc and S. this agent induces potentially significant respiratory depression (ABOU-MADI 2001). IPSAV. USA) and the arterial partial pressure of carbon dioxide (PaCO2) in isoflurane-anaesthetized birds of prey. Y.Service de médecine zoologique. ranging from 416 g to 2062 g. Thus. Nine birds. DMV. Desmarchelier. and fast inductions and recoveries. Fitzgerald. Nellcor Puritan Bennett. Plesanton. 1 INTRODUCTION Isoflurane is the most commonly used inhalant anaesthetic in birds. two barred owls (Strix varia) and a great-horned owl (Bubo virginianus) were used for this study. The significant dissimilarities between the avian and mammalian respiratory physiologies prevent cross-species extrapolations. the pulmonary parenchyma of birds is not formed of alveoli as in mammals but of parabronchi. For instance. in such a way that the partial pressure of carbon dioxide of the air exiting the pulmonary parenchyma is higher than the PaCO2 (SELLER 2000) Obviously. a Cooper’s hawk (Accipiter cooperii).samples is the gold standard to assess the balance between CO2 production and elimination. 2001). Canada) was administered IM prior to mask induction with isoflurane (AErrane®. such as the NBP-75® (Nellcor Puritan Bennett. Plesanton. PETCO2 measured with these monitors might provide a sufficiently accurate estimation of PaCO2 (CASATI et al. USA). In mammals. Furthermore. This project was carried out according to animal utilization protocols approved by the Faculté de médicine vétérinaire animal care committee which operate under the auspices of the Canadian Council on Animal Care. Each bird was intubated with a Magill uncuffed endotracheal tube (diameter: 3 to 4. Nellcor Puritan Bennett. two rough-legged hawks (Buteo lagopus). the relation between PaCO2 and PETCO2 will be affected by this physiological feature. EDLING et al. 2002. 0. Baxter. CA. Therefore.5mm) connected to the semi-open non-rebreathing Mapleson D modified Bain circuit. Toronto. Heart rate was monitored with an ultrasonic Doppler (Ultrasonic Doppler Flow Detector. Wyeth Animal Health. Université de Montréal. end-expired partial pressure of carbon dioxide (PETCO2) is widely used as an indirect non-invasive alternative to monitor the ventilatory status during general anaesthesia (O’FLAHERTY 1994). these birds were euthanased at the end of the experiment. 2 MATERIAL AND METHODS Three red-tailed hawks (Buteo jamaicensis). To reproduce anaesthetic protocols used for orthopaedic procedures commonly performed in raptors. 1059 ± 459 g). Gas exchange occurs in a cross-current system that increases O2 uptake and CO2 release. 2000). The objective of our study was to evaluate the usefulness of a handheld Microstream capnograph (NPB-75®. since most species of birds seen by veterinarians have relatively small tidal volumes and high respiratory rates. measurement of PETCO2 in these species with traditional high flow (150 ml/min) side-stream capnometers designed to be used in humans or in domestic mammals will most likely result in erroneous readings. USA) to assess the PaCO2 and PETCO2 relationship in different species of birds of prey anaesthetized with isoflurane using a Bain non-rebreathing system. surrounded by air capillaries. Very few studies have explored the usefulness of PETCO2 as a parameter to evaluate arterial CO2 dynamic in birds (PETTIFER et al. Unfortunately. Plesanton. Due to permanent injuries that prevented their release. were admitted in the rehabilitation program of the Raptor clinic of the Faculté de médecine vétérinaire. ON. Guelph. which weighed from 416 g to 2062 g (mean ± SD. Parabronchic air flow is unidirectional during both expiration and inspiration. These birds.5mg/kg of butorphanol (Torbugesic®. Canada) delivered in oxygen. Microstream capnometers. arterial blood sampling is not practical in most bird species. have a much lower aspiration rate (50 ml/min). ON. CA. model 811- 212 . Nova Biomedical. arterial blood pressure. Boucherville. 30.5 ml of arterial blood was collected from the arterial catheter and injected back into the intraosseous catheter. Parks Medical Electronics. PaCO2. PETCO2 values below and above the dashed line respectively overestimate and underestimate the concomitantly measured values of PaCO2. USA) applied on the palatine artery. individual differences between measured PETCO2 and PaCO2 were quite variable.95 . Canada) (NICHOLSON et al. NPB-75®. Fig.1) and between PETCO2 and arterial pH (r2 = 0. Statistical analysis were performed using a mixed multiple regression model with individual as a random factor to correlate PETCO2. Birds were euthanased at the end of the procedure with an intravenous injection of T-61® (Euthanasia solution.5% to 4% with a fresh gas flow of 0.97 . Dashed line is the line of equality for comparison. p < 0. Inc. then 0.. Due to spontaneous breathing. 3 RESULTS AND DISCUSSION We collected 38 analyzable samples from the nine birds used for this study.8 L/min.3 mg/kg atracurium (atracurium besylate®. The arterial catheter was connected to a three way valve which allowed blood pressure monitoring between arterial blood collections. An intraosseous catheter was inserted into the tibiotarsus using a proximal approach. weight.0001) (Fig. 1992). Strong correlations were observed between PETCO2 and PaCO2 (r2 = 0. PETCO2 levels were first stabilized near each target level for five minutes. Approximately 0. QC. USA) (KILEY 1979). Intervet. and the setting of the isoflurane vaporiser. arterial pH. Canada).0001). five of the nine birds had to be paralyzed with an intravenous or intraosseous injection of 0. Whaltam. p < 0. Blood gas analysis was performed within 5 minutes of sample collection with values normalized to body temperature according to human normograms associated with the blood gas analyzer (Stat Profile M®. Birds were manually ventilated with a peak inspiratory pressure varying between 15 and 20cm H2O. The sampling tube of the capnograph (Handheld capnograph. Ventilation rates were modified to reach different levels of PETCO2. Whitby.4 ml of arterial blood was then sampled from the arterial catheter in a heparinised 1 ml syringe. 50 and 60 mm Hg. A 24-Gauge catheter was inserted in the superficial ulnar artery to facilitate repeated arterial blood collections. 40. However. Isoflurane was delivered in oxygen at a concentration ranging from 2. Arterial blood was sampled for five levels of PETCO2 : 20. Pleasanton. 213 . ON. Sabex. USA) was connected to a paediatric T-piece adapter located between the endotracheal tube and the Bain circuit. Nellcor. Continuous line represents the best-fit linear trend line.B. MA. OR. Aloha. 1: Scatterplot of PaCO2 against PETCO2 in mmHg (n=38). CA. This is contrasting with results encountered with the mammalian alveolar system where the PETCO2 is usually lower then the PaCO2 by 2 to 5 mm Hg due to the presence of alveolar dead space or V/Q mismatch. Another potential explanation for this finding might be the voluminous dead-space of the avian respiratory system. The low respiratory rates needed to obtain elevated PETCO2 (one PaCO2 (mmHg) 214 . the error reported for the NPB-75® Nellcor capnograph for values between 39 to 99 mm of Hg is 5%. Indeed. analytic errors do not account. 1. All but one of the PETCO2 levels measured above 53 mm Hg underestimated the concomitantly measured PaCO2 level (mean ± SD.5 mm Hg higher then their paired PaCO2 (mean ± SD.5 ± 5.0 mm of Hg). the difference between PETCO2 and PaCO2 varied from -7 to 13 mm of Hg. However. at least entirely. the parabronchi and aerial capillaries arrangement enables a cumulative transfer of CO2 from the venous blood to the parabronchic lumens resulting in a PETCO2 higher than the PaCO2 (FEDDE 1986). Measured levels of PETCO2 ranging from 39 and 53 mm Hg were an average 1. for the differences observed between PETCO2 and PaCO2 during hypercapnia. 5. Within this range.2 mmHg). This result is similar to what was reported for African grey parrots (Psittacus erithacus timnus) anaesthetized with isoflurane using a modified Norman elbow non-rebreathing system and receiving intermittent mechanical positive pressure ventilation.7 ± 3. which is much lower then the discrepancy observed.80 75 70 65 60 55 50 45 40 35 30 25 20 15 10 10 15 20 25 30 35 40 45 50 55 60 65 70 PETCO2 (mmHg) All but one of the PETCO2 levels measured below 39 mm Hg exceed the concomitantly measured values of PaCO2 by 3 to 11 mm of Hg (mean ± SD. This overestimation of PaCO2 by PETCO2 is in agreement with the effect of the cross-current exchange system in the pulmonary parenchyma of birds. These results suggest that the accuracy of the capnograph used in this study decreases with high values of CO2. -8.5 mm Hg). Consequently.8 ± 6. CASATI A et al. NICHOLSON A et al.833. In : SEYMOUR C and GLEED R.293. In: STURKIE PD. FORBES NA.626. This study shows that. respiratory rates) were correlated with the difference between PETCO2 and PaCO2. SELLER TJ. weight. 8. PETTIFER GR et al. 2. we proposed that anaesthetized birds should be ventilated in order to maintain levels of PETCO2 between 30 and 45 mm Hg. it is possible that passive mixture occurred between CO2-rich post-parabronchic gas and gas present in the air sacs. None of the other parameters tested (isoflurane concentration. 1994.1. O’FLAHERTY D. EDLING TM et al. arterial partial pressure of O2 and O2 saturation of haemoglobin remained within normal limits throughout the procedures. Avian anesthesia. 16: 268 . 6. 17: 622 . 9. Capnography – Principles and practice series. Accuracy of end-tidal carbon dioxide monitoring using the NBP-75 microstream capnometer. Vet Clin N Am: Exot Anim Pract 2001. Shurdinton: BSAVA. This shows that these parameters are not sensitive indicators of the ventilatory status in anaesthetized birds receiving 100% inspired oxygen.breath every 1 or 2 minutes) were probably associated with a transient stagnation of gas exiting the pulmonary parenchyma. despite some individual variations. European J Anesthesiology 2000. Interestingly. 1999: 283 . Birds. Based on the results obtained in the current study. Respiration and cardiovascular responses to exercise in the duck. arterial blood pressure. 1.2342. J Appl Physiol 1979. Boca Raton: CRC Press 2000. 219: 114 . 3. FEDDE MR. KILEY JP et al. Bird respiration vol. as a result. 4. 47: 827 . A study in intubated ventilated and spontaneously breathing nonintubated patients. 7. Therefore. 4: 147 . resulting in a dilution of the CO2 concentrations and. 1986. Avian Physiology. 10. the monitoring of PETCO2 with a side-stream Microstream capnometer provides a sufficiently accurate estimation of PaCO2 in isoflurane-anaesthetized birds over 400g receiving manual positive ventilation with a Bain system. The comparative cardiopulmonary effects of spontaneous and controlled ventilation by using the Hallowell EMC anesthesia workstation in Hispaniolan Amazon parrots (Amazona ventralis). BMJ Publishing Group.167. 215 . Manual of small animal anaesthesia and analgesia. 53: 2337 .1718. Am J Vet Res 1992. 4 CITATION INDEX ABOU-MADI N. J Avian Med Surg 2002. 5. Neuromuscular and cardiovascular effects of atracurium in isoflurane-anesthetized chickens. London. J Am Vet Med Assoc 2001. Capnographic monitoring of anesthetized African grey parrots receiving intermittent positive pressure. Respiration. an artefactually low PETCO2 at the following expiration. New York: Springer-Verlag.276. Université de Montréal C. Faculté de médecine vétérinaire de
[email protected]. QC J2S 7C6. DMV.AUTHOR ADDRESS M. IPSAV Service de médecine zoologique. Desmarchelier. 5000. St-Hyacinthe.fr 216 . Canada Email: marion. University of Heidelberg. Austria. It is an important health problem in Psittacines. Haefeli2. M. W. followed by A. The mean Cmax after IV application of 10mg/ kg voriconazole was ~7. Enteral absorption of the drug was low with a calculated bioavailability of <20% after PO application. University of Veterinary Medicine. that would indicate organ damage. During treatment with 10mg/kg PO for 30 days no clinical signs of side effects or disease have been observed. Burhenne2. as well as in some zoo-birds. Different antimycotics have been used for treatment of birds (OROSZ and FRAZIER 1995. itraconazole is currently considered as the first choice for treatment of aspergillosis in birds (OROSZ and FRAZIER 1995.Birds ABSTRACT Voriconazole (Vfend®. Clinical Pharmacology and Pharmacoepidemiology. Reptile and Fish Medicine. No relevant changes of clinical chemistries were seen. E.Clinic for Avian1. Scope1. FISCHER and HATT 2003). J.Antifungal agent – Pharmacokinetics . the half-life was about 2 hours in chicken. Because of its broad spectrum and low incidence of toxicity. There are some concerns that this drug may cause severe side effects like 217 . INTRODUCTION Aspergillosis is a frequent cause of respiratory disease in birds. Department of Internal Medicine VI2. evaluated in chickens as a model. Hess1 KEYWORDS Aspergillosis – Voriconazole . Germany PHARMACOKINETICS AND PHARMACODYNAMICS OF THE NEW ANTIFUNGAL AGENT VORICONAZOLE IN BIRDS A. highly potent triazole antifungal agent with high in vitro activity against a wide variety of fungal pathogens. raptors and waterfowl maintained in captivity. after PO application <1 µg/ml. The most common etiologic agent is Aspergillus fumigatus. 1. 1995). LUMEIJ at al. The present study was performed to gain information on pharmacokinetics and safety of voriconazole in birds.5µg/ml. flavus and A. Vienna. niger. Pfizer) is a novel. There were no remarkable differences of plasma concentrations of voriconazole between single applications and long-term treatment indicating that there was no accumulation of the drug. Detection was performed either by UV detection or mass selective detection (LC/MS). All other study parts were performed with the 10 mg/kg dose which clinical evidence suggests is effective in aspergillosis treatment. highly potent triazole antifungal agent that is a derivative of fluconazole with high in-vitro activity against a wide variety of fungal pathogens.5. Plasma samples were spiked with internal standard and borate buffer was added. 1. Each group consisted of three chickens to receive 5. 10. 218 . The analytical methods were validated according to FDA validation guidelines and fulfilled all quality assurance for accuracy. In this species it is recommended to switch to other drugs like ketoconazole or amphotericin B.0. Limits of quantification were 0. including Aspergillus. 2. robustness. or 15 mg/kg voriconazole PO.205/192-BrGT/2003 and consisted of three parts. The linear calibration range was 0.20)µg/ml up to 10. MATERIALS AND METHODS 2.5.99.CNS symptoms. QUESENBERRY et al. The extracts were dried. and even death in African grey parrots (VANDERMAST et al. ESPINEL-INGROFF et al. Performance of the study parts The study was conducted under animal testing permit No. 0. 6. One case of a cockatiel with suspected aspergillosis that has been treated with voriconazole has been reported (LANGHOFER 2004). (ABRAHAM et al. 1999. 2002). This new antifungal appears therefore a promising potential alternative for the treatment of aspergillosis in birds. 2. Plasma concentrations were evaluated after 0. 7. but no standardized studies on pharmacokinetics have been published yet. Analytical Methods An analytical method for the determination of voriconazole in plasma was developed utilizing high pressure liquid chromatography (HPLC) and solid phase extraction (SPE) technology. 1. 1991). Correlation coefficients were always r2>0. GZ 68.20µg/ml (HPLC/UV) and 0. reconstituted and analyzed by HPLC. Voriconazole (Vfend®. and 12 hours. Pfizer) is a novel. vomiting. 2002.05 (0. but with the drawbacks of other possible side effects or lower efficacy.0µg/ml. especially in African grey parrots. 1990. 2. Candida. The present study was performed to gain first information on pharmacokinetics and safety of voriconazole in birds using the chicken as a model. 2001). and recovery. precision. OROSZ and FRAZIER 1995. and Cryptococcus spp. These samples were loaded onto SPE columns and eluted with organic solvent. 3. In humans this drug has been efficacious in treating invasive aspergillosis and is well tolerated (DENNING et al.05µg/ml (LC/MS). Part 1: For the evaluation of single dose pharmacokinetics and dose dependency single doses of voriconazole administered PO to four groups of chicken. MUIJSERS et al. CK. Part 3: The pharmacokinetics and safety of long-term application of voriconazole was studied in a group of 9 birds receiving 10 mg/kg voriconazole PO for up to 30 days. 1. GLU. 1.0 hours and were used to calculate bioavailability. Plasma concentrations were determined after 10. time to reach Cmax (tmax). 0. 3. From the same samples.25 ± 0.15 Part 2 Pharmacokinetics after IV applications are shown in Table 2.0. GLDH. maximum plasma concentrations (Cmax). TP. 20.25. Plasma concentrations were determined in blood samples taken after 0. and 3. LDH. oral bioavailability was less than 20%. CA. In a randomized cross-order design. URIC) were also determined for safety assessment. Table 2: Intravenous pharmacokinetics of 10 mg/kg voriconazole in chickens. and the half-life (t1/2) of voriconazole. plasma biochemistries (AST. Dose [mg/kg] 5. Parameter IV Part 3: Plasma values at treatment days 10.1 AUC [(µg/mL)*h] 1.Part 2: For the assessment of the bioavailability of voriconazole in chicken two groups of two chickens were assembled.7 ± 0. and 30 at 1 hour and 3 hours after application were always lower than 3µg/ml and showed no meaningful increase with the application duration. each chicken got 10 mg/kg voriconazole PO or IV separated by a washout phase of 1 week.75.4 ± 0. 2.7 Cmax [µg/mL] 0. ChE.0 .7 tmax [h] 1. AUC0-3h [(µg/ml)*h] ~11 VD [ml/kg] <1700 Clt [ml/(h*kg)] <600 t1/2 [h] 1. 20. and 30 days 1 hour and 3 hours after dosing to monitor whether drug accumulation occurs.50. Due to the very fast elimination (t1/2~2 h) in chicken only low Cmax values were reached and the AUC values showed no significant increase with the dose. 0. RESULTS Part 1 Table 1 shows the single dose pharmacokinetic of part 1.3 ± 0.5. No clinically relevant changes of clinical chemistries occurred during the observation period.0.2 219 . Table 1: Mean values of pharmacokinetic parameters in chickens. 5. MANAVATHU EK. RIBAUD P. 5. 34: 563 . VANDERMAST H. Emerging antifungals and the use of voriconazole with amphotericin to treat Aspergillus. GOA KL and SCOTT LJ. et al. BOYLE K and SHEEHAN DJ. 7. 1. 150: 101-15. J Assoc Avian Vet 1991. Antifungal agents: A review of their pharmacology and therapeutic indications.5µg/ml suggesting that antifungal concentrations throughout the dosing interval may be insufficient for some fungi. 220 . 5th ECAMS Scientific Meeting.22. Voriconazole: in the treatment of invasive aspergillosis.4. itraconazole and amphotericin B. Roundtable discussion: Clinical therapy. New Orleans 2004: 21 .191. 2001) and depends on the infectious agent. 8. Mycopathologia 2001. Efficacy and safety of voriconazole in the treatment of active aspergillosis. FISCHER I and HATT J-M. CUTRIGHT JL and CHANDRASEKAR PH.2664. Preliminary results of voriconazole in Psittacine species indicate that there might be big species-specific differences in pharmacokinetics. MUIJSERS RB. CITATION INDEX ABRAHAM OC. 10. In vitro antifungal activities of voriconazole and reference agents as determined by NCCLS methods: Review of the literature. Tenerife 2003: 21 . DENNING DW. 4: 189. Mycology 1999. Proc Assoc Avian Vet.05-2µg/ml voriconazole for Aspergillus spp. DORRESTEIN GM and WESTERHOF J. SPEER B and FLAMMER K. ESPINEL-INGROFF et al. OROSZ SE and FRAZIER DL. Plasma and tissue concentrations of itraconazole in racing pigeons (Columba livia domestica). 33: 7 .11. 9: 8 . BAUCK L.571. J Avian Med Surg 1995. This is mainly caused by the short half-life of the compound in chickens and the poor oral bioavailability. Drugs 2002. 1999. GORGEVSKA D and WOESTENBORGHS R. J Assoc Avian Vet 1990. In vitro susceptibilities of Aspergillus species to voriconazole. ESPINEL-INGROF A. A fatal treatment of sinusitis in an African grey parrot. J Avian Med Surg 1995. Pfizer.18. Clin Infect Dis 2002. USA). 9. MILPIED N. 4. The poor bioavailability together with the rather small VD (limited tissue distribution) might indicate that effective therapies would require multiple dosing to reach sufficient plasma concentrations. Antifungal therapy in birds: A review and critical discussion of new trends. LANGHOFER B. QUESENBERRY K. 62: 2655 . LUMEIJ JT. This is in obvious contrast to the very good absorption of voriconazole in humans (bioavailability up to 96%). The manufacturer recommends 0. 6. 9: 32-5. 5: 186 .24. The fact that even during treatment with 10mg/kg PO for 30 days no evidence for adverse effects or organ damage emerged may at least suggest that it does not have substantial toxicity. DISCUSSION The minimum inhibitory concentration (MIC) of voriconazole has been evaluated in vitro (ABRAHAM et al. as is the fact that during longterm administration no accumulation occurred. 2. Only for a short period of time this limit has been exceeded after IV application of 10 mg/kg voriconazole and after PO application the mean Cmax was even <0. for humans (Vfend® information sheet. Because only relatively low plasma concentrations were achieved a final assessment of the safety of the compound is not possible. 3. NY. at 221 . University of Veterinary Medicine. 1210 Vienna. Reptile and Fish Medicine.AUTHORS ADRESS Alexandra Scope. Austria Email: alexandra. Veterinärplatz 1.ac. DVM Clinic for Avian.scope@vu-wien. Two birds with severe aspergillosis died 22 and 30 days respectively after the end of treatment. negative in 4/23. 222 . but more often from 21 to 30 days.Psittacus erithacus . but did not fully recover because of chronic lesions on 2/23. No negative clinical effects were recorded in 20/23 birds. Ages varied from 3 months to 42 years. gastro-enteritis and loss of condition. but in African grey parrots this drug is reported to be toxic. vomiting and yellow or dark faeces.African grey parrot . An alternative treatment is fluconazole. suspension at 10mg/ml) was administered at 15mg/kg q12h. dermatitis.M. The 24 birds examined consisted of 19 Psittacus erithacus and 5 Psittacus erithacus timneh. Aspergillosis serology was performed by electrocineresis on 23/24 cases. Thirteen parrots were treated with fluconazole PO as the sole antifungal agent and 10 with a combination of fluconazole PO and amphotericin by nebulisation. Positive clinical results were noticed in all birds: 18/23 recovered. old age on 1/23. The duration of treatment varied from 5 days to 75 days. The effects were reported by the owners. severe disease and death later on 2/23.Fluconazole . A retrospective review of 24 African grey parrots treated with fluconazole PO allowed us to assess the efficiency and tolerance of the protocol used. A positive result was obtained in 19/23 cases. No difference in negative clinical effects was seen between fluconazole alone or associated with amphotericin. Of the remaining. one case (during prolonged treatment) reported decreased appetite.Clinique vétérinaire Sigean. France CLINICAL ASSESMENT ON THE USE OF FLUCONAZOLE PER OS IN 24 AFRICAN GREY PARROTS (PSITTACUS ERITHACUS) : ACCEPTANCE. Pericard DVM KEYWORDS Birds . SIDE EFFECTS AND EFFICIENCY. The birds presented with a range of clinical symptoms including upper and lower respiratory disease. the 5/23 remaining cases improved. One of them reported tiredness during treatment. These deaths reflect probably more a lack of treatment and the severity of the disease than a toxicity of the drug.Electrocineresis Aspergillosis ABSTRACT Itraconazole is the more commonly used antifungal agent in birds. J. The fluconazole (Triflucan TM. Treatment was well accepted in 23/24 cases and refused in only 1/24 cases. further observations were made by the author during consultations. positive clinical effects.1 INTRODUCTION Fungal diseases commonly occur in birds. This is the case of injectable antifungal drugs. a higher incidence of aspergillosis is seen in the African grey parrot (Psittacus erithacus). suspension at 10mg/ ml). combination of amphotericin by nebulisation. or timneh). duration of treatment with fluconazole. doses per day. the assessment on the use of fluconazole PO in the African grey parrot is important. 223 . 2. 2 Treatments Thirteen parrots were treated with fluconazole PO as the only antifungal agent. Ages varied from 3 months to 42 years. The 24 birds examined consisted of 19 Psittacus erithacus and 5 Psittacus erithacus timneh. The birds presented with a range of clinical symptoms including upper and lower respiratory disease. and duration between beginning of treatment and last contact. The history of the selected cases was then studied for the chosen data: age. serology of aspergillosis by electrocineresis. subspecies. Unfortunately. Many factors are known to favour aspergillosis. dermatitis. symptoms. Among psittacine birds. gastro-enteritis and loss of condition. In addition. the treatments of fungal diseases have been improved with the advent of new drugs and better knowledge of the bioavailability in certain avian species. The duration varied from 5 to 75 days. 1 Cases review The review was carried out by selecting cases of our database. The effects were observed by the owners and during consultations. Due to this. but more often 21 to 30 days. negative clinical effects. b) treatment: including fluconazole. weight. including the injectable forms of itraconazole and amphotericin. based on 3 criteria: a) species: African grey parrot (either nominal. During the last few years. c) date: from 1st January 2003. 10 with a combination of fluconazole PO and amphotericin by nebulisation (suspension at 0. 2 MATERIAL AND METHODS 2. but in that species immunosuppression due to a poor nutrition and husbandry may play the most important role. The dose used was 15mg/kg q12h (Triflucan TM. African greys do not tolerate this drug very well and may show signs of toxicity at standard therapeutic doses (OROSZ et al 1996. Aspergillosis of the respiratory tract is common but systemic infections such as fungal dermatitis and fungal disease of the gastro-intestinal tract are also commonly seen. itraconazole seems to be the preferred choice against Aspergillus. some drugs or galenic forms are not readily available to veterinarians in France. number of administration per day. was contacted by phone to complete the history if needed. if not seen at the clinic recently. Every owner.5mg/ml). POWERS personal communication). Among these drugs. 2 Clinical effects No clinical negative effects were noticed in 20 on 23 birds treated with fluconazole. The bird received fluconazole during a total of 75 days on 115 days. CPK 648 UI). Nutrition was very poor. total protein 38g/l. a pelleted food and improved. 1 Acceptance Among these 24 African Grey. It received fluconazole during more than 21 days. At serology the bird was positive for aspergillosis and chlamydophilosis. Periods of 21 days were separated by 7 to 13 days without treatment. A positive result was obtained in 19/23 cases. 3. The bird not tested was a chick with crop fistula under short preventive treatment. The only case (No 2450) in which side-effects were noticed was a very old bird (40 years) with weight loss and less active. arthrosis. Paris) on 23/24 cases. probiotics. Other treatments included doxycycline PO in water. 3 RESULTS The cases are listed in Table 1. injection of vitamin A. the owners reported that the administration was easy. but also general improvement. pelleted food and fresh fruits. Other treatments include nebulisation with amphotericin at 0.5mg/ml. modified vocalisations for 3 years. The first case (No 3788) had a recurrence infection with severe chronic air sacculitis. This bird was not handled by their owners and did not try again! For the rest. diarrhoea. Two birds died later. The blood chemistry was within reference values (uric acid 39mg/l. The bird died suddenly 2 months after the beginning of fluconazole (and about 1 month after the end of fluconazole). but these were birds with severe disease.2. The treatment with fluconazole as the sole antifungal agent was prolonged because the owner noticed a significant improvement. ASAT 129 UI. The owner did not call us and no necropsy was performed. then nothing during 10 days and then again 10 days of fluconazole. and because it was considered that a single 21 days treatment was not sufficient. The owner noticed tiredness. a poor nutrition. This serology is used as a diagnostic tool. 3. only one refused to take the drug. and to monitor the recovery by repeated samples. glucose 2. probiotics. The owner did not wish a necropsy. The bird died 22 days after the end of fluconazole. polyuria and loss of balance. 3 Serology of aspergillosis Aspergillosis serology was performed by electrocineresis (Laboratory Ceri. The second case (No 4312) was also affected by aspergillosis with weight loss. calcium 101mg/l. negative in 4/23. It received fluconazole during 36 days.35g/l. One owner preferred to give it mixed with jam instead of liquid. The bird has a feather picking syndrome for years. and later doxycycline. juvenile formula and pelleted 224 . Doses and frequency of fluconazole treatment The doses and frequency we chose for most of these treatments were 15mg/kg q12h (RUPIGER unpublished. Eleven were treated with fluconazole alone. 2 Clinical improvement The other 5 birds of the 23 treated improved. bile acids 56. During the second period of 21 days of fluconazole. weight gain. 3. The 2 others were considered only improved because the opacity of air sacs remains. The blood sample for chemistry taken 2 days after showed decreased total proteins (uric acid 57mg/l. However. able to fly and later able to fly without dyspnoea. reduction of polyuria (No 4312). better appetite (No 3788). The clinical signs did not reappear after a short treatment with clomipramide and lespedeza extracts. 4. improvement was noticed: able to speak again. Many authors think that for aspergillosis the duration must be at least several weeks and 225 . 1 Clinical recovery Recovery was seen by disappearance of the symptoms in 18 of the 23 birds treated with fluconazole. 4 DISCUSSION 4. Duration of treatment The duration of treatment is chosen according to the disease and results. Even for the 2 who died later. Our results show a good clinical efficacy and few side effects at this dose.2). the owner noticed yellow stools and decrease appetite.food. The symptoms disappeared when the treatment with fluconazole stop and were not present at the time of the consultation 2 days after (no vomiting. 3 Positive clinical effects 3. in OLSEN and OROSZ. No doxycycline was given at that time. normal stools). more active. total protein 25g/l. 7 with a combination of fluconazole and amphotericin in more severe cases.1µmol/l). this cannot stand in for the lack of studies on the bioavailability of fluconazole in the African grey. better vocalisation. The treatment with fluconazole was discontinued when the bird started vomiting and passing dark stools. one of them is still under treatment. “méconnaissable” (No 2450). 3. 2000). 2 days after the beginning of a new period. The one who had side-effects also improved considerably. The negative results of the electrocineresis for aspergillosis and the duration between the treatment and the last contact were also good indicators of recovery (see table 1). 3. limited by its old age: behaviour more active. 3. 2. LDH 257 UI. a lot higher than 20mg/kg q48h or 10mg/kg q24h as proposed by FLAMMER (1996). There were no statistical differences between the side-effects during a treatment with fluconazole alone or associated with amphotericin (P=0. 1. ASAT 135 UI. calcium 91mg/l. It seems as good as treatments with itraconazole Per Os in experimental aspergillosis: 41/60 quails survived in the itraconazole treatment group. Symptoms disappeared immediately when the treatment stop and before consultation. 4. 0/60 in the untreated group (GUMUSSOY et al. Side-effects and toxicity Fluconazole has negligible hepatic metabolism. and another one was noticed to have a better appetite! The effects observed on case No 2450 (lower appetite and yellow stools. In this case review. Because amphotericin also has a renal metabolism (OROSZ 2003). only one of 23 had a transient lower appetite. then vomiting. Centre de Pharmacovigilance Veterinaire de Lyon. and has a neutral pH. Then the potential toxicity is very different from itraconazole. we made interruption of about 10 days between duration of 3 to 5 weeks. is probably due to a selective bias: more severe cases were treated with the association of antifungal agents. while itraconazole has extensive hepatic metabolism. 4. In the present cases. It is water soluble and well absorbed PO. dark stools. less severe ones with fluconazole alone. Ecole Veterinaire de Lyon. 226 . 2004). 3. but we could not confirm this. 5 ACKNOWLEDGEMENTS I thank Dr Florence Buronfosse. Fluconazole preferentially accumulates in the kidneys and is excreted in the urine. 21/23 survived) is very good. cumulative toxicity could be possible when fluconazole is used at the same time. But it is not easy to separate the effects of the treatments and of the disease itself! Some African greys are reported partially to totally anorexic during fluconazole treatment (OROSZ 2003). ANDRE 2004). It is not linked to proteins contrary to itraconazole. The idea is to minimise the possible toxicity by accumulation (see under). The apparent better results of the fluconazole alone compared to fluconazole and amphotericin. and hypoproteinemia) might be linked to renal insufficiency. For longer treatments. for her help and advice with this study and Dr Glen Cousquer for kindly reviewing the manuscript. well tolerated by mucosa (review in OROSZ 2003). 4. Urine analysis was impossible in that bird since it had a large amount of mixed stools due to a dilated cloaca.sometimes several months (review in ABUNDIS-SANTAMARIA 2002. Efficiency The global results of the treatments (18/23 recovered. We share that opinion and often made treatments of about 1 month. no statistical difference was seen on side effects and deaths. ABUNDIS-SANTAMARIA E. St Louis: Mosby.L. ANDRE J.pdf. 2. (ed): Mexico: Cervantes Olivares.350.M. La Teste 33 France. AUTHORS ADRESS J.E. Experimental Aspergillus fumigatus infection in quails and results of treatment with itraconazole. 6.173. 490. Published by the author.C..E.J. AGUILAR R. ATASEVER A. Péricard. and OROSZ S. SCHROEDER E. FRAZIER D. Tampa 1996: 203 .uk/secure/veterinary/ Aspergillosis. 2004.H. 5.P. 3.
[email protected]. CES de pathologie aviaire. J Vet Med B Infect Dis Vet Public Health. 10: 168 . 7. Vet Clin Exot Anim 2003.pericard. 2004. De la maladie à la bonne santé.204 4. Unpublished data in OLSEN G. 51(1): 34 . et al. Proc Assoc Avian Vet. Oiseaux de cages et de volières.E.aspergillus.. 2000.6 CITATION INDEX 1. DVM. FLAMMER K. 2002: 31. OROSZ S. 6: 337 . Aspergillosis in birds of prey. OROSZ S. www. Antifungal drug therapy in avian species.: Manual of Avian Medicine. diplôme d’épidémiologie Clinique vétérinaire. RUPIGER D. GUMUSSOY KS.man. Pharmacokinetic properties of itraconazole in blue-fronted Amazon parrots (Amazona aestiva aestiva) J Avian Med Surg 1996. CAM Y. rue du Cers.fr 227 . France Email: jm. Fluconazole in psittacines birds. UYANIK F. 1113 Sigean. per day Negative clinical effects Positive clinical effects Serology electrocineresis aspergillosis Last contact after treatment (month) 8 24 4 15 19 4972 4? 512 Regurgitation 4233 2 422 4246 5. thinness 0 arc then 2 then 1 3 4429 18 471 Nasal granuloma.2 2 Yes None >30 21 25 30 2 2 Yes No None None 3788 18. but chronic lesions. loss of voice. air sacculitis Rhino-sinusitis. low appetite 36.7 2 Yes None Recovery 23. can speak.7 2 Yes None Recovery 27.3 2 No 30 2 No 29. rhinosinusitis. voice loss.3 2 No None Gain of weight Recovery 1 arc then 0 1 arc then 0 1 arc then 0 2 arcs then 0 1 arc Weight (g) Symptoms Duration fluconazole days Dosis mg/kg per day Nb of admin.5? 356 Recurence of aspergillosis with severe chronic air sacculitis. loss of voice.Table 1. chlamydophilosis Improvement but chronic lesions 1 arc then 0 19 « « 1 arc 1 arc then 0 22 16 4636 0. prostration.7 2 Yes None None None Tiredness Died 22 days after end of fluconazole Improvement: voice. gout Air sac opacity Loss of voice. nasal granuloma. control after purchase Improvement but chronic lesions Gain of weight Recovery .air sacculitis. +gout. prostration.2 2 No None Recovery 35. Results of the clinical assessment on the use of fluconazole PO in 24 African Grey parrots (Psittacus erithacus) No Ampho Age (years) 23 30 31 30 15 31. stop 10 10 30 2 Yes 33 21? 21 29. bullet & ancient fracture.7 410 Thinness 4754 5? 477 Vomiting. prostration 4361 ? Adult 448 Severe dyspnea.5 538 4451 3 398 4484 12 256 Dyspnea. sinusitis 1 arc then 0 1 arc then 1 then 0 1 arc 19 16 14 4639 0. Recovery Recovery Recovery 228 >60 31.4 480 Thinness.2 2 Yes None Recovery 29. 4313 3.9 1 Yes 23.25 488g Dyspnea during effort. polydypsia. «asthma» Dyspnea.8 1 Yes 26 2 No None None None None None Recovery Improvement then Recovery Recovery Recovery Improvement Recovery 27 2 No None 29 2 No None 36 2 No None Recovery Recovery Recovery Recovery 30 2 Yes None Recovery « 10. then 0 « 10 15 8 8 15 5 21 21 30 not given 30 35 37. anorexia. prostration. dark green stools. less active . reduction of polyuria Improvement « « « « 21. polyuria. anorexia. nervous troubles Crop fistula 2565 42 368g Diarrhea.stop 10 30 2 No Improvement: behaviour. keratitis. anorexia. low appetite Vomiting.. loss of voice Prostation. flight… Improvement: flight without dyspnea. 15 30 30 30 2 2 2 30. stop 7 30 2 No yellow stools. automutilation. thinness « 4165 ? Adult 496 Almost dead (abandonment).3 2 10.4312 21.. conjonctivitis. diarrhea. voice. less active None but sudden death 2 months later None Improvement: gain of weight. taeniasis « 560 5000 1.75 423 5293 1. loss of voice 4198 12. prostration. stop 10 2.4 24 19 4787 ? 375 4964 8 275g Timnhe 229 4694 0.5 305g Timnhe Dermatitis.5 4 13 9 0. dark stools 4 0 arc 0 arc no test 0 arc 0 arc 1 arc 25 1 arc then 1 1 arc then 0 1 arc then 0 1 arc then 0 2 arcs then 1 0 then 1 then 1 then 0 25 9 5 1.stop 13 21.1 1 No No Yes Yes No No None None None 11. stop 7.2 2 No 2 arcs 2 372 Loss of weight. stop 30 2 No Vomiting.6 232g Timnhe 5457 14 259g Timnhe 710 35 390 Prostration. trouble of equilibrium 2450 40 427 Loss of weight. sinusitis. air sacculitis Loss of weight. muscle development.5? 421 Rhinitis. decrease appetite 30 2 No 2 arcs then 3 ? Adult >21 32. rhinitis. C. 1999. is a NSAID that has recently become available in the U. LVT. Hernandez-Divers. DACZM. 1 INTRODUCTION Reduction of pain and inflammation is an important part of medical and surgical management of avian patients. Dipl ABVP (avian). but this method has been fraught with potential complications in the past. the studies that have been performed in birds are predominately in species not commonly kept as companion animals.5mg/kg. (BAERT et al. The results suggest that meloxicam should be administered at 0. which has been used in veterinary medicine for many years in Europe. DVM. Meloxicam (Metacam. Peak serum concentrations were 1. Merial. and M. POULSEN et al.2mg/kg PO q 24h.S. and ducks. ostriches. K. (BUDSBERG et al. While NSAIDs have been used and studied in many mammalian species. MRCVS. Frequently. DVM. 2002. K. DACVP. University of Georgia. quail.Psittacula krameri ABSTRACT This study evaluated the pharmacokinetics of meloxicam given to ring-necked parakeets (Psittacula krameri) orally and intravenously at a dose of 0. 1996. 1991) Furthermore. 1998. LEES et al. drug dosages for birds are extrapolated from mammalian dosages. there is little data on their use in birds. The elimination half-life was 16 hours after oral administration and 4 hours after intravenous administration. such as poultry. Grant. PhD. it has only been licensed for use in dogs. Duluth. Meloxicam has been shown to have nearly 100% bioavailability when administered orally with food in dogs. Budsberg. 230 . DACVS. TURCK et al.86ug/ ml after oral and intravenous administration respectively.College of Veterinary Medicine. S. H. United States of America PHARMACOKINETICS AND USE OF MELOXICAM IN PSITTACINE BIRDS G. GA USA). S. Wilson. Latimer. S. for which the reported dosage is 0. BUSCH et al.5mg/kg twice a day if given orally or six times a day if given intravenously. BVetMed.1 to 0. Pethel KEYWORDS Meloxicam .DZoo Med.Pharmacokinetics . At this time.Ring necked parakeets . DVM. 2002.38ug/ml and 3. were used.5mg/kg twice a day orally or every 4 hours intravenously is recommended in psittacine birds. Bioavailability was 100% for intravenous administration and was also high for oral administration.BAERT et al. The pharmacokinetics of meloxicam have not been established in any parrot species.0mg/kg orally q 12h. the purpose of this study was to establish the pharmacokinetics of meloxicam given orally and intravenously and to evaluate bioavailability in one species of psittacine birds. Therefore. and pigeons. 2 MATERIAL AND METHODS Twenty adult ring-necked parakeets (Psittacula krameri) of both sexes. BAERT et al. When meloxicam was given to these birds in the injectable form. 231 . turkeys. Plasma concentrations of the drug were determined by highperformance liquid chromatography methods. the shorter the half-life. have not looked at oral bioavailability. (BAERT and DEBACKER 2003) In humans and rats and presumably in birds. ostriches. although there are anecdotal reports of its usage in pet birds in Europe at a dose of 0.3 to 1. Results showed that the half-life of meloxicam in ring-necked parakeets was of longer duration than in any of the previously studied non-psittacine avian species for both the oral and IV routes. the previous studies in chickens. No more than 10% of the estimated total blood volume for any individual was removed within a 24 hour period. Based on these results a dose of 0. 2003.5 mg/kg was given intravenously (IV) to each of the 20 birds. Pharmacodynamic studies are needed to further understand the disposition of meloxicam in psittacine birds. The collections times for each bird were randomized to minimize the effect of iatrogenic anaemia on drug dosage concentrations and health of the animal. there were significant differences in elimination rates of the drug between different species.5 mg/kg) administered orally (PO). 2002. weighing between 110 and 140 grams each. Blood samples were again taken from each bird at several times over a 24 hour period.(VELPANDIAN et al. GRAHAM et al. so the larger the bird. A dose of 0.5mg/kg given orally or IV. based on the results of this study and previous studies in other species of bird. 3 RESULTS There were no apparent clinical side effects noted in the birds in this study at a dose of 0. After a two week washout period the subjects then received the same dose (0. meloxicam is eliminated partly in urine and partly in faeces. 2001) Also. ducks. 2000) The UGA Institutional Animal Care and Use Committee approved the protocol. Blood samples were taken prior to administration of the drug (time 0) and from each bird again at two of several randomized time samples over a 24 hour period. it appears that the half-life is inversely proportional to the weight. 4 DISCUSSION Interestingly. JAISWAL J. 5. University of Georgia. BAERT K and DEBACKER P. 16(2): 123 129. QUANDT JE. 10. HIGGINS AJ. et al. Proc Assoc Avian Vet. VELPANDIAN T. DVM. J Avian Med Surg 2002. BUSCH U. 449 . Pharmacokinetics and pharmacodynamics of meloxicam in the horse. BAERT K and DE BACKER P. Development and validation of a new high-performance liquid chromatographic estimation method of meloxicam in biological samples. Heather Wilson. 738(2): 431 . BHARDWAJ RK and GUPTA SK. SEDGWICK AD. Orlando 2001: 19 . Disposition of sodium salicylate. flunixin and meloxicam after intravenous administration in broiler chickens. ABVP-avian Department of Small Animal Medicine. A review of the clinical pharmacokinetics of meloxicam. et al. flunixin. 9. Pharmacodynamic and pharmacokinetic aspects of the non-inflammatory non-steroidal agent meloxicam in dogs. Pharmacokinetics of ketoprofen in adult Japanese quail. NACKAERTS J and DE BACKER P. 106(3): 94 100.6. 147(2): 97 .33. Athens. TURCK D. BAERT K. Pharmacokinetics of meloxicam in animals and the relevance to humans. GA 31522. 6. HEINZEL G. Drug Metab Dispos 1998.21. TELL LA. AUTHORS ADRESS G.5 CITATION INDEX 1. Disposition of sodium salicylate.453. LEES P. Dtsch Tierarztl Wochenshcr 1999. ROTH W and BUSCH U. Comparative pharmacokinetics of three non-steroidal anti-inflammatory drugs in five bird species. Comp Biochem and Physi 2003. 4. GRAHAM JE. BUDSBERG SC. 2. KOLLIAS-BAKER C and CRAIGMILL AL. and meloxicam after intravenous administration in ostriches (Struthio camelus).edu 232 . J Vet Pharmacol Therap 2002. 7.63.436. College of Veterinary Medicine. J Chromatogr B Biomed Sci Appl 2000. Am J Vet Res 2002. 134(1): 25 . 35(1): 13 . Br Vet J 1991. POULSEN NB and HORSTERMANN D. CROSS AR. 26(6): 576 . Evaluation of intravenous administration of meloxicam for perioperative pain management following stifle joint surgery in dogs. 25.uga.108. et al. United States of America Email: hwilson@vet. SCHMID J. 63(11): 1557 . Dipl. Br J Rheumatol 1996. 8. 3.584. E. Nevertheless. Heart glycosides (digoxin. several studies about the examination of cardiac function have been done over the last years (KRAUTWALD-JUNGHANNS et al. 1999. Germany THE USE OF ENALAPRIL IN BIRDS : INDICATIONS. digitoxin) have a positive inotropic effect on the contraction of cardiac muscle. 1995. Kuhring. 2004). K. BOSKOVIC et al. In contrast. PEES et al. WILSON et al. but also possible side effects of the ACE-inhibitor enalapril in birds. it is possible to assess cardiac function and to diagnose alterations intra vitam. 1 INTRODUCTION Recent studies show the incidence of cardiac disease as a common finding in post mortem examinations (KRAUTWALD-JUNGHANNS et al. Krautwald-Junghanns. intra vitam diagnosis is comparably rare due to the non-specific clinical signs and accompanying diseases overlying the cardiovascular symptoms. today. DrMedVet. Unfortunately. PHARMACOKINETICS AND POSSIBLE SIDE EFFECTS M. ProfDrMedVet. First experiences indicate the high potential value.Enalapril ABSTRACT Cardiac disease in birds occurs more frequently than suspected previously. With the ability to diagnose cardiac alterations ante mortem. CLINICAL EXPERIENCES. the state of scientific knowledge is far below that of mammal medicine. Pees. The only group of cardiac drugs for which pharmacokinetic data is available for birds are heart glycosides (HAMLIN and STALNAKER 1987. the possibility of an efficient therapy becomes more important. Only few studies and some case reports about cardiac therapy in birds exist.Clinic for Birds and Reptiles University of Leipzig. DiplECAMS KEYWORDS Birds – Cardiac Disease – ACE-inhibitor . electrocardiography (ECG) and especially echocardiography have been found to be of diagnostic value. improve 233 . 1989). Radiology. CARRANI et al. 2004). M. concerning this point. 2003. With the ongoing technical progress and increasing experience. parasitological and microbiological examination. Only healthy birds without signs of any disease were used for the study. Enalapril was given over a period of 21 days with regular control examinations (body weight. 0.2. and 24. 12.0h. necropsy was performed to evaluate the cause of cardiovascular disease and accompanying diseases. 16. enalapril tablets (Enacard® Tablets. 2. Clinical experiences In our clinic.0h. their main indication is stabilizing birds in emergency situations.5h. Therefore ACE-inhibitors lead to an improvement of renal function.0h.0h. enalapril was administered orally to 15 pigeons. and long-time application seems to be critical in birds due to difficulties in controlling effect and plasma levels. blood 234 . The birds were housed in single caged and fed with a commercial seed diet and water ad libitum. 2 MATERIAL AND METHODS 2. Blood was taken from the birds and examined (WBC.0h. the effects of the administration on the clinical picture. In our experience. Germany) were dissolved in sterile water and applied with a crop cannula following body mass. Intervet.and afterload of the heart. For the examination of the pharmacokinetics. Their mechanism is based on the inhibition of the conversion from angiotensin I to angiotension II. 39 adult pigeons were included in the study. For the examination of dosis-dependent side-effects. their therapeutic margin is very small. Since the half-life period and effectivity of enalapril is higher compared to captopril (SPONER 2002). For the application. 2. they underwent a thorough clinical. 4. clinical examination. an increase of diuresis and a decrease of blood pressure resulting in a decreased pre. Unfortunately.1. In mammals. enalapril) are frequently used for therapy of cardiac problems in human and mammal medicine. The aim of this study was to summarize clinical experiences with this drug in birds. Blood was taken on defined time intervals (0h. 2001). blood chemistry) after an acclimatisation period of 7 days. 24 pigeons were divided into groups with each 8 animals. this drug is preferable for use in birds. and to give first guidelines for the application and the control of the therapeutic success.0h. Before administration of enalapril. 6.0h) to check for drug concentration. enalapril is used for the therapy of cardiovascular disease since 4 years. the survival time and side-effects that are potentially related to the drug administration have been noted. Pharmakokinetics and dosis-dependent side effects To examine the pharmacokinetics and the dosis-dependent effects of enalapril. In birds that have died. the therapeutic margin of enalapril is not as critical as the margin of cardiac glycosides (SPONER 2002).0h. ACE-inhibitors (captopril. Besides clinical examination. radiography was performed and the heart was examined by means of electrocardiography and echocardiography. 1. This enzyme is responsible for the contraction of the vessels and the retention of sodium and water in the kidneys.its relaxation and decrease the heart rate. 8. During this period. Dosage was 5 mg/kg body mass. Sudden death has been reported after some days of application (PEES et al. liver congestion and free fluid in the thoraco-coelomic cavity. Ideopathic hydropericardium was diagnosed several times and also treated. one group was treated with 5 mg/kg orally once daily. dosing and application was easier than with tablets. not as tablets but as solution of the tablets in water.8% (day 1) to 36. It seems that the taste of the solution is favoured by the patient since the acceptance is better than the acceptance of pure water or any other drug used in the clinic. In consequence of this good acceptance most owners could apply the drug regularly and without problems. the wall thickness of the interventricular septum and the contractility of the ventricles was measured by means of echocardiography (values see table 1). The effect on the cardiac function could be proven with a long-term therapy in an amazon bird suffering from right-sided heart failure. 3 RESULTS AND DISCUSSION 3. and electrocardiography). Clinical experiences The main indications for the use of enalapril in birds were alterations of the right ventricle (hypertrophy. too. insufficiency of the muscular atrioventricular valve) and following congestion into the large circulatory cycle (resulting in liver congestion. There is nothing known if this depends on the drug itself or on accompanying ingredients of the tablets. Concerning the application. This improvement was always connected with an improvement of the status of the cardiovascular system. With this solution.examination. In birds with high-grade pericardial effusion and/or ascites. The improvement of cardiovascular function could be seen in a decreased congestion. coordination) could be seen in the majority of the treated patients. Correspondingly. an improvement of the general body condition (activity. a regular observation was that the acceptance of this drug was extraordinary well. the drug was administered orally. appetite. and also the final decompensation (day 825) could be seen (decrease of contractility of both ventricles). in birds that not reacted to the enalapril administration. there was also no change in the cardiovascular function. As in mammals.6% (day 140)). hydropericardium and ascites). visible in a reduction of pericardial effusion. One group was treated without application of enalapril (placebo). An improvement of the cardiac function (increase of the fractional shortening (FS)of the right ventricle from 26. a left ventricular dysfunction was diagnosed and treated with enalapril. Diagnosis was made in all cases by means of ultrasound (after preceding hints for cardiovascular disease from the clinical examination and/or radiology). The clinical use of enalapril over the last 4 years indicates a broad safety margin of this drug in birds. Concerning the effect of enalapril on the cardiovascular system and the general health status. dilatation. and the third group was given 10 mg/kg orally once daily. echocardiography. In this birds the size of the ventricles. the use of enalapril was combined with the surgical removal of fluid from the thoraco-abdominal/pericardial cavity by means of ultrasound-guided abdominocentesis/ pericardiocentesis 235 .1. Only in one bird. the long term application of enalapril should be monitored carefully with regular blood examinations. 2. J Avian Med Surg 1995.11 PEES M. Transcoelomic two. that was possibly caused by an increased diuresis. STRAUB J and KRAUTWALD-JUNGHANNS ME. Admittedly it has to be said that the diagnosis of lowgrade heart alterations is still difficult and based on individual experience rather than objective measurements. 27: 334 . et al. In these cases. 6. 148(17): 540 .und Nachtgreifvögeln (Accipitriformes. J Vet Pharmacol Therap 1987. Pharmacokinetics and dosis-dependent side effects Results of the examinations concerning the pharmacokinetics and dosis-dependent side-effects will be presented at the conference (ongoing research). 3. 4. 236 .2. the therapy with enalapril (and additional therapeutic measures) could improve the life quality and the survival time of the birds (up to 2 ½ years). 2003). The remaining fluid could be resorbed after administration of enalapril (in some cases in combination with furosemide 0. Strigiformes). HAGNER D. et al.31.341. 3. 9: 19 . the reduction of fluid from the pericardial cavity seemed to have an influence on the cardiac function (decreased pressure on the heart). Falconiformes. Vet Rec 2001. Insufficiency of the muscular atrioventricular valve in the heart of a blue fronted amazon (Amazona aestiva aestiva). 145-149. et al. SCHULZ M. KRAUTWALD-JUNGHANNS ME. Tenerife 2003. On the other hand. In birds with high-grade pathological alterations of the heart. clinical experiences show that with dosages of 1 mg/kg/day. The only side effect that has been observed clinically during long-term therapy in dosages of 5 mg/kg/day orally was an increase of the PCV. Basis for use of digoxin in small birds. BOSKOVIC M. STALNAKER PS. GELLI D. In birds with idiopathic signs of congestion or only low-grade alterations cardiac function seemed to be normal after the signs of congestion had vanished. Proc Eur Assoc Avian Vet. CARRANI F. KRAUTWALD-JUNGHANNS ME. KRAUTWALD-JUNGHANNS ME.356. SALVATORI M. BRAUN S. A dosis reduction after improvement of the general health status is advisable. Since this dehydration was severe. 18(1): 2 . J Avian Med Surg 2004. PEES M. FAILING K.543.1 to 2 mg/kg/day). Practical cases will be presented at the conference.dimensional echocardiography in the avian patient. 10: 354 . Research on the Anatomy and Pathology of the Psittacine Heart. Möglichkeiten und Grenzen echokardiographischer Untersuchungen bei Tag.(STRAUB et al. HAMLIN RL. Tierarztl Prax 1995. et al. also the effectivity on the cardiovascular systems seems to be reduced. 4 CITATION INDEX 1. A preliminary echocardiographic initial approach to diastolic and systolic function in medium and large parrots. 5. PEES M. An den Tierkliniken 17. 04103 Leipzig. ZENOBLE RD.uni-leipzig. PEES M. 10. Echocardiographic examinations of 60 African grey parrots and 30 other psittacine birds. 9. Stuttgart: Enke 2002: 147 .434. HORTON CR. Löscher W (eds): Lehrbuch der Pharmakologie und Toxikologie für die Veterinärmedizin.76 8. DrMedVet. ENDERS F and KRAUTWALD-JUNGHANNS ME. STRAUB J.169. 20: 432 . 155: 73 .de 237 . Pharmakologie des Herz-Kreislauf-Systems. WILSON RC. AUTHORS ADDRESS M. Germany Email:
[email protected]. Vet Rec 2004. et al. J Zoo Wildl Med 1989. ResECAMS Clinic for Birds and Reptiles. Single dose digoxin pharmacokinetics in quaker conure. In: Frey HH. SPONER G. University of Leipzig. Pericardiocentesis and the use of enalapril in a fischer’s lovebird. Vet Rec 2003. Pees. STRAUB J and KRAUTWALD-JUNGHANNS ME. 152: 24 . Therefore the aim of this study was to develop an oral therapeutic regime and to determine an appropriate dosage of doxycycline with optimal effectiveness and lower side effects for treating chlamydophilosis in pet birds using the racing pigeon as a model. 2 MATERIAL AND METHODS For all studies. is still common.Doxycycline . POWERS 1997).Chlamydophilosis . E. that doxycycline is the drug of choice to treat the clinical signs and to handle the elimination of the pathogens. The birds had free access to 238 . They were housed in individual cages with a wired floor at room temperature and an artificial daylength of 16 hours. F.Oral administration ABSTRACT Chlamydophilosis in racing pigeons. Weilacher. Dipl ECAMS. adult. R. There have been several studies about the possible administration of doxycycline (PRUS 1992. Prof DrMedVet.Antibiotic therapy . Krautwald-Junghanns.Pigeons . In avian medicine doxycycline is known especially for its effectiveness in treatment of Chlamydophila infections. healthy racing pigeons (Columbia livia domestica) of both sexes were used. Ungemach. as in any pet bird species. In recent years this drug regained importance again because of its effectiveness against a broad antibacterial spectrum with good compatibility and low costs. University of Leipzig. BIOAVAILABILITY AND COMPATIBILITY OF DOXYCYCLINE IN BIRDS AFTER ORAL ADMINISTRATION E.Clinic for Birds and Reptiles. M. Germany PHARMACOKINETICS. Prof DrMedVet KEYWORDS Birds . 1 INTRODUCTION For decades doxycycline is a frequently used antibiotic in veterinary medicine. Investigations by several authors show. M. In the last section (4) the doxycycline tolerance was examined over 20 days after oral administration. Considering the half-life of doxycycline the drug was distributed in two equal doses twice a day and administered with a crop cannula into the crop. 16. 6. The voluntary intake of doxycycline medicated drinking water and the resulting doxycycline plasma levels were tested. Blood samples were taken from the ulnar vein at various times before and after administration of doxycycline. one group with 60mg/kg/d and the fifth group with 150mg/kg/d. one group was treated with the therapeutic dose of doxycycline of 30mg/kg/d. 2. The blood and organ concentrations of doxycycline were determined with HPLCanalysis with UV-detection after reconditioning the samples extracted by solid phase extraction.5. 12. 1. half of the animals one week later. 24. 7 and 13. Blood was taken at defined intervals after the application (0. Half of the pigeons were necropsied the day after the last application. 4. 2. 48. This was done to check for effective doxycycline concentrations in the organs needed for therapy of Chlamydophila infections. 239 . Blood samples were taken and examined (haematology.water and a standard breeding grain-mix for racing pigeons. blood chemistry. On the basis of these results in the following section 2 36 racing pigeons were examined over 14 days. Doxycycline blood concentrations were determined of expected peak and trough values on day 2. Sixty pigeons were divided into groups of 12 animals each. For this purpose a single dose of doxycycline of 60mg/ kg was administered to 15 pigeons via crop cannula. 6. microbiological and parasitological). food and water consumption were recorded. At day 1. 40. Section 3 was conducted to simulate the conditions in the field. 0. 72 h) and examined for doxycycline concentration. 36. No grit-mineral mixture was given during the experiments. doxycycline concentration). The study was divided into four sections: The aim of section 1 was to assess whether a sufficient doxycycline absorption in the gastrointestinal tract after oral administration had been achieved and to examine pharmacokinetic parameters. The main question was whether a reduced dose of 30mg/kg/d maintains constantly high and effective plasma levels for treating Chlamydophilosis. one group with 45mg/kg/d. 10 blood samples were collected two times a day (8 am and 4 pm) and the doxycycline concentrations determined. 4. 8. The doxycycline solution was administered divided into two equal dosages with a crop cannula. Before and during the study the pigeons were checked for their health status (clinical. After 14 days of doxycycline administration levels in lungs and liver of 12 pigeons were examined 2 and 5 hours after the last doxycycline application. One group was treated with a placebo (0mg/kg/d). Twenty racing pigeons received doxycycline medicated drinking water at a concentration of 750mg/l ad libitum for 10 days. After 20 days a pathological examination was performed. 8. 6 7.7 18.4 6. t1/2 = 11.5 h.1µg/ml.3 184. Table 1. Pharmacokinetic parameters after a single application of 60mg/kg doxycycline Parameter Cmax Tmax �Z t1/2 AUC0-∞ MRT0-∞ CL/f VZ/f Unit [µg/ml] [h] [h-1] [h] [µg·h/ml] [h] [ml/min·kg] [l/kg] Section 1: single application of 60mg/kg BW 8.3 l/kg.0 h.0615 11.1 6.4 5.5 and 2. VZ/f = 5.3 In section 2 (oral doxycycline administration) all doxycycline blood levels were above 1 µg/ml.8 5. Table 2. The determined values fluctuated between 1.3 h.0554 12.4 (peak in the steady state) 1.7µg/ml.2 [h] [µg·h/ml] [h] [ml/min·kg] [l/kg] 240 .0 0.1 18.3 RESULTS The evaluation of section 1 (checking for pharmacokinetic parameters) resulted in the following average values: cmax = 8.8 (trough in the steady state) steady state 0.5 37. this resulted in a calculated elimination half-life of 12. necessary for treating chlamydophilosis. Doxycycline was well absorbed in the gastrointestinal tract of pigeons after oral administration. tmax = 6. Doxycycline plasma concentrations could be measured up to 73 h after the last application. Pharmacokinetic parameters after application of 30mg/kg (2x 15mg/kg) doxycycline for 10 days Parameter Cmax Tmax �Z t1/2 AUC0-∞ MRT0-∞ CL/f VZ/f Unit [µg/ml] [h] [h ] -1 Section 2: twice a day application of 15mg/kg BW 2. Some birds failed to maintain doxycycline concentrations above 1µg/ml for the entire period. 4 DISCUSSION Due to practical considerations and despite known possible disadvantages the application of antibiotics via drinking water is preferred. The organ/plasmalevel indicated an accumulation of the highly lipophilic doxycycline in these organs. respectively 4. PADILLA (2003) reported comparable data in fruit doves. However. The organ/plasma-level of the doxycycline concentrations were about 8 for the liver and about 2 for the lungs and also confirmed an accumulation of the high lipophilic doxycycline in the organs of racing pigeons. blood chemistry. The pigeons of this group showed regurgitation after administration of the drug. after 14 days of doxycycline administration reached 14.5fold. This offers a possibility to avoid production of medicated feeding stuff that is often not very well accepted or injections that may produce muscle necrosis. Under field conditions with a voluntary intake of drinking water a good acceptance of doxycycline medicated drinking water was found. The results of our studies show that doxycycline is well absorbed in the gastrointestinal tract of pigeons after oral administration. the results of the group with the 5fold therapeutic dose differed significantly from the other groups.62 up to 2. FLAMMER (2000) in psittacines. for food and water intake. Furthermore. At a concentration of 750mg/l of 241 . and the 2fold therapeutic dose were observed by the clinical examination.The doxycycline levels in liver and lungs. A loss of bodyweight was registered. haematology and at necropsy. In the blood samples taken two times a day (8 am. but the majority of birds reached or exceeded these levels during treatment. the group receiving the therapeutic dose. In pet bird medicine this is for example especially important in racing pigeons to keep the birds in good flight condition. They had polyuria and yellowish uric acid. the mean daily doxycycline dose ingested was 27mg/kg. the 1.8µg/g 5 hours past the last doxycycline application. GGT. bile acids and cholinesterase were significantly increased during the application period. Section 3 showed a good acceptance and voluntary intake of doxycycline medicated drinking water and constant doxycycline plasma levels for 10 days. In accordance with an average intake of medicated drinking water of 18. The water intake increased from an average of 20 ml to an average of 50 ml/pigeon.4 and 2. Microscopically an extramedullar haematopoesis (2 pigeons).20µg/ml were detected.0ml/kg/d. 3 pm) average doxycycline concentrations of 1. Pathological gross findings were diffuse yellow-brown or yellow-red coloured livers. In section 4 no significant differences between the placebo group. high doxycycline levels observed not only in the plasma. Constant doxycycline blood levels were achieved exceeding 1µg/ml needed to effectively treat chlamydophilosis. AST. At any time the concentrations were above 1 µg/g.7µg/g 2 hours past the last doxycycline application.3 and 9. but also in the target and elimination organs for Chlamydophila (lungs and liver). acute hepatitis (1 pigeon) and fatty liver cell degeneration (1 pigeon) was seen. Higher doses should be avoided. On the other hand the pigeons which received the 5 fold therapeutic dose showed side effects like regurgitation and loss of body weight. MILLER RE and FLAMMER K. because they could induce liver damage and therefore impair the well being of the animals.doxycycline in the drinking water again plasma levels above 1 µg/ml were reached. Germany Email: weilacher@vmf. Proc Am Assoc Zoo Vet. Doxycycline in drinking water for treatment of Chlamydophila psittaci in fruit doves. FLAMMER K: Preliminary notes on treatment of chlamydiosis with doxycycline medicated water. PRUS SE.268. This fact has to be considered especially in diseased birds with a higher water intake. This is in accordance with other studies (PADILLA 2003). Proc Assoc Avian Vet. PADILLA LR. 5 CITATION INDEX 1. 4. Dosing methods for administration of doxycycline in cockatiels. It can be assumed that the tolerance of doxycycline for racing pigeons after oral administration for 20 days is excellent up to the twofold therapeutic dose. CLUBB SL and FLAMMER K. Avian Dis 1992. Portland 2000: 3 – 5. liver damage with following polydipsia. Minneapolis 2003: 267 .uni-leipzig. Proc Assoc Avian Vet.de 242 .5 fold and 2 fold therapeutic dose. polyuria. 2. yellowish colouring of the uric acid and increased liver enzymes. Doxycycline plasma concentrations in macaws fed a medicate corn diet. AUTHORS ADRESS E M Weilacher.483. 480 . POWERS L and FLAMMER K. 3. Reno 1997: 57 – 58. DVM Clinic for Birds and Reptiles University of Leipzig An den Tierkliniken 17 04103 Leipzig. In section 4 no side effects of doxycycline for the therapeutic dose and with an application period of 20 days could be found. In contrast to studies in psittacines however (FLAMMER 2000) we did not see any side effects in racing pigeons for the 1. a clinical effectiveness could not be postulated for the treatment with chloramphenicol against a S. Schmidt Res ECAMS and N. the clinical efficacy of chloramphenicol for treatment of a S. typhimurium infection. coli strain that proved to be pathogenic for pigeons rsp. coli infections in racing pigeons in a dosage of 12. the treatment with different modes of oral application was started. E. coli infections (study 1) rsp. Both furazolidone preparations proved to be effective in treating gastrointestinal E. The pigeons originated from conventional breeders and were housed in different groups (control -.Clinic for Birds and Reptiles University of Leipzig. After this time. By legal 243 . 24 adult pigeons. typhimurium infections (study 2) was investigated by examination of 36 rsp. The negative identification of the marked bacteria was determined as main parameter for the clinical efficacy of the treatment. typhimurium var. a S.E. however best results were obtained by application via capsule for 5 days. Reitz KEYWORDS Furazolidone – Chloramphenicol .racing pigeon ABSTRACT In two similar studies. This phase was followed by an adspectory phase and/or by a pathological examination in study 2. After infection with an E. In Germany. Copenhagen strain. the animals developed clinical signs of disease.5 mg /pigeon. the clinical efficacy of furazolidone for treatment of gastrointestinal E. coli – Salmonella . Particularly minor species are concerned by that. animals. V. capsule and powder group). whose number is too small to animate pharmaceutical enterprises to investigate the costs of the admission. 1 INTRODUCTION Changes in drug regulations and requirements often cause an omission of different effective drugs from the market. Germany ORAL FURAZOLIDONE AND CHLORAMPHENICOL FOR TREATMENT OF GASTROINTESTINAL BACTERIAL INFECTIONS M. Krautwald-Junghanns. ProfDrMedVet.Oral application. In contrast to study 1. furazolidone and chloramphenicol are allowed only for veterinary purposes and here only for racing pigeons. DiplECAMS. contact dermatitis. Furazolidone is called a “cavity therapeutic agent”. 1996). However when furazolidone and chloramphenicol are considered. The advantage of the bacteriostatic furazolidone is its broad antibacterial spectrum and the insignificant formation of resistance. This was due to the potential side effects chloramphenicol may cause in men like aplastic anaemia. typhimurium var. The veterinary importance of chloramphenicol is based on its broad antibacterial spectrum and a slow development of resistance. since its therapeutic effect is only in the intestine and it is hardly absorbed from there. Campylobacter. Drugs used Furazolidone: In 1995 the use of nitrofuranes in food-supplying animals was already forbidden in all member states of the EU (KROKER et al. in poultry the double therapeutic dose can lead to cardiomyopathy (KROKER et al. Mycobacterium and Pseudomonas possess a natural resistance (LÖSCHER et al.. Copenhagen using different oral modes of application. In this context it was the aim of our study. In racing pigeons this antibiotic is mainly used to treat samonellosis caused by Salmonella enterica subsp. 1996).. but many other sensitive strains like Klebsiella.conversion they may be used also for other pet and zoo birds. Due to the bad systemic compatibility after parenteral application furazolidone has to be administered locally or orally (KROKER et al. These bacteria rank among the most frequent causes of bacterial diseases of the avian intestinal tract. The permission is limited to the treatment of bacterial gastro-intestinal tract infections caused by E. Chloramphenicol may penetrate the cell wall. coli infection of the gastro-intestinal tract. to examine the clinical effectiveness of furazolidone and chloramphenicol as chemotherapeutic agents in gastro-intestinal tract diseases caused by E. Salmonella and Shigella may be covered as well (KROKER et al. Waterfowl may react particularly sensitive to furazolidone and should therefore not be treated with this antibiotic drug (GYLSTORFF and GRIMM. 1998). enterica Serovar Typhimurium 244 . Due to its lipophilic effect it may be used to treat bacterial central nervous diseases. they have to be used under critical consideration among others of their side effects. which is mainly plasmid bound. Nitrofuranes have a small therapeutic margin. so it is effective against intracellular bacteria. Chloramphenicol: Already in 1990 the use of chloramphenicol was forbidden in foodsupplying animals in all member states of the EU. It was predominantly used therefore in former times in poultry for the treatment of infectious enteritis. coli for furazolidone and those caused by Salmonella typhimurium for chloramphenicol. 1996). S. Other side effects that have been described may be regurgitation in pigeons after oral application and bone marrow depression in embryos (so chloramphenicol should not be used in breeding hens). Besides that the nitrofuranes possess a spermicidal effect. Another important point for the mode of application of these drugs is their half-life and whether sufficient blood and tissue levels may be achieved for example by the common oral application. 2002). coli rsp. The effectivity spectrum of the bacteriostatic chloramphenicol covers many sensitive strains of most gram positive and gram negative bacteria. 1999). so their application is contraindicated in breeding animals. The resistance situation of nitrofuranes is generally good. For the pigeon however there are only little current publications. Main indication is the E. Capsule: The daily dosage was one capsule/ pigeon. typhimurium var. powder) were infected with a marked. However the half-life of chloramphenicol in pigeons reaches only 0.capsule and powder group). so that each pigeon got (average weight 500g) daily 12. which are used in practice for oral administration over the drinking water or instilled into the crop. The controlled clinical studies were performed as parallel’s group study. After oral administration the absorption is almost complete. the animals developed clinical signs of disease. Copenhagen (called S. coli. 1992). 2 METHODS The studies were performed according to valid regulations as well as according to the principles of the “Good Clinical Practice “ (GCP). The furazolidone powder contained 500 mg furazolidone per 7. Then. Directly before the application of the furazolidone a solution of the powder with warm water was prepared in each case.var.FURAZOLIDONE Dosage and application Two different furazolidone preparations were applied (powder and capsule form). coli strain could be re-isolated from the cloaca of all pigeons. coli strain (infection dose 1 x of 109). The negative identification of the marked bacteria was determined as main parameter for the clinical efficacy of the treatment. The apparent distribution volume is very high. Copenhagen strain.once in the morning and once in the evening. 2. The pigeons originated from conventional breeders and were housed in different groups a 12 birds (control . drinking water). Powder: Since the drinking water admission is dependent on numerous factors in pigeons. After infection with an E.26 h (CLARK et al.5 mg furazolidone per capsule. a S. pathogenic E. After completion of the therapeutic part on day 5 and 6 swabs were taken from choana. coli strain that proved to be pathogenic for pigeons rsp. the animals got a more exact dosage into the crop 2 x daily every 12 hours . 245 . The theoretical therapy dose was 25 mg/kg BW furazolidone. started the therapy for 5 days as described. 1999). After this time started the treatment with different mode of application (capsule. The elimination is predominantly renal (LÖSCHER et al. inoculated. Twenty four hours after the inoculation the marked E. these were dipped directly before the installation into paraffin oil.5 g packing unit according to manufacturer data in powder form. crop and cloaca and examined for the presence of the inoculated E. Like that an application volume of 20 ml/ kg BW of the accordingly medicated drinking water was instilled in each pigeon by crop cannula. typhimurium in the text). 1 STUDY 1 . Expiration of study 1 The pigeons of the two therapy-groups (capsule. the application of the capsules took place once daily in the morning one hour before feeding. In order to ensure a better admission of the capsules.5 mg furazolidone over 5 days. the furazolidone in capsule form contained 12. This phase was followed by an adspectory phase and – in case of the Salmonella infection – by a pathological investigation. 2. Since the drinking water admission is dependent of temperature. Expiration of study 2 The pigeons were infected orally with a chloramphenicol sensitive.CHLORAMPHENICOL Dosage and application The chloramphenicol powder used was certified for oral administration in the drinking water and contained (according to manufacturer data) per 6. Other side effects could not be found. clear signs of clinical disease could be found and all animals shed the inoculated S. Directly before the application a solution with a concentration of 16. The re-isolation was performed due to described methods after euthanasia of the animals from the individual organs. Copenhagen strain. 3 pigeons in the capsule group showed regurgitation. The dosage was determined on the basis of the following formula: 650 mg chloramphenicol /kg BW/day x BW (kg) of the animal treated mean daily drinking water admission (l)/ animal =. infect. STUDY 2 . In the adspection and clinical investigation already 24 hours after beginning of treatment. In the powder group this improvement was seen approx. 48 hours after beginning of therapy. 246 .5g packing unit in powder form in powder form 850 mg of the active substance cloramphenicol. The infection dose was 1x109. However a positive tendency of this clinical criterium (towards physiological faeces) was already seen in the capsule and powder group from day 5 on. Copenhagen strain. so that the therapeutic part of the study began. 2.. Only after completion of the therapy phase a normalization of the faeces in the two treated groups could be seen. pathogenic S. typhimurium var.Results of study 1 At day 5 and 6 in none of the treated pigeons the marked “E” could be isolated in contrast to the birds of the control group. the animals were treated – in contrast to study 1 and due to the bad results of various pre-studies with lower application intervals .. 3 to 7 days p. mg chloramphenicol per l water The middle daily drinking water admission was determined with 40ml/kg BW. Due to the orange colour of the regurgitated material it was most likely that this contained the dissolved capsules. typhimurium var. The application intervals of the drug had to be shortened to 3 hours (see above) and the application duration was extended to 10 days.25mg chloramphenicol/ ml was made with warm water in each case. At the last day of drug application. a clear improvement of almost all investigation criteria could be seen in the capsule group. air humidity and other factors.five times daily (every 3 hours) with an application volume of 20ml/kg medicated drinking water by crop cannula. the 12 animals of the untreated group were S. The clinical improvement of the treated pigeons however could not be brought in connection with an efficient destruction of the pathogen. an improvement could be documented in the phase of acclimatisation (infected but not treated). taste of the drug and actual diseases may play an important role in this context. behaviour.5mg/ pigeon. body attitude and plumage in the treated groups. typhimurium. typhimurium strain could be re-isolated from all pigeons from the faeces. all reisolated S. coli infections in racing pigeons in a dosage of 12.26 hours and the peculiarities of Salmonella infections this result is explainable. kind of food. Due to the short half-life of chloramphenicol in racing pigeons of 0. 3 DISCUSSION Both furazolidone preparations proved to be effective in treating gastrointestinal E. typhimurium positive as well. Both in the treated as well as (as expected) in the untreated group pathological-anatomical findings in the liver could be found. In order to ensure the admission of an effective dose in the two studies an application with crop cannula was necessary instead of the application via drinking water. Concerning the oral modes of application in these studies and in the field: Since the housing conditions in the single cages in an air-conditioned room do not correspond to the actual housing conditions in the field (with free flight etc. The latter does not apply to furazolidone and chloramphenicol. For the treatment with chloramphenicol it has to be noticed in summary that the main criterion for the evaluation of the clinical effectiveness of chloramphenicol in treating S. As expected. Interestingly most of these alterations were seen in the pathological-anatomical investigation immediately after the end of treatment (day 10). Significant differences in the isolation from the single organs could not be determined. typhimurium strains were chloramphenicol sensitive. however best results were obtained by application via capsule for 5 days. After the therapy with chloramphenicol.). at the beginning and at the end of the therapy in the following points. Due to the inaccurate dosage with this kind of application however actually only preparations should be used. 1998). on day 10 and 16 organ samples were taken. In the daily adspection and the clinical investigations. for an antibiotic treatment of larger bird collections the antibiotic is usually applied within the drinking water despite the well-known disadvantages just due to practicability reasons. were so far not observed in the pigeon. Under practice conditions without doubt substantially larger fluctuations in the actually intake of drinking water have to be expected: Thus among other things hot weather conditions. typhimurium positive. Generally. however incompatibilities as described for water fowl (GYLSTORFF and GRIMM. In contrast to the latter.Results of study 2 6 days after the inoculation the inoculated S. typhimurium infection was not fulfilled. a strongly reduced drinking water admission under these condition was expected due to various previous experiences. 247 . which have large therapeutic margin. The inoculated S. which hint at an infection with S. an exact dosage per animal could be achieved with the crop installation like by capsule application. Nevertheless when these drugs are administered via drinking water the actual seasonal drinking water intake of the individual bird group should be considered in each case. all 12 animals of the treated group were S. typhimurium strain could be re-isolated in all pigeons of the treated group. . Tagungsberichte der 2. Possibly the noticed temporal difference lies in the high concentration of the active substance in the capsule. As a further disadvantage it is described that the pigeons may vomit the capsules/ tablets. UNGEMACH F. A clear improvement of the clinical symptoms occurred quite early (as opposed to the powder group) in the capsule group .. 2. UNGEMACH F. H.. R. and W.de 248 . 5. 2. that even under natural conditions with free access to the medicated drinking water. 4 CITATION INDEX 1. Another point that has to be considered is. University of Leipzig. without this is noticed by the owner.. 7.): Lehrbuch der Pharmakologie und Toxikologie für die Veterinärmedizin. 4. KROKER.uni-leipzig. pees@vmf. Stuttgart: Enke Verlag. KROKER R. Berlin: Blackwell Verlag 1999: 243 . LÖSCHER W.uni-leipzig. a sufficient effective blood level may not be kept constant.): Lehrbuch der Pharmakologie und Toxikologie für die Veterinärmedizin. (eds. W.H. 2002: 387 . Stuttgart: Ulmer Verlag 1998: 148.. R. 3. R. Leipzig: Leipziger Samstagsakademie 2003.501. KROKER. ProfDrMedVet. Nitrofurane. In: FREY. In: FREY.244 KROKER.besides the unusual application by crop cannula . In our own study.-H.. Stuttgart: Enke Verlag 1996: 499 . since particularly in the night the pigeons do not drink and thus a sub-therapeutic effect may develop. Antibiotikatherapie bei Brieftauben.the most exact possibility of the oral administration of a drug in birds. Germany Email: krautwald@vmf. Vogelkrankheiten.already 24 hours after beginning of therapy.LÖSCHER (eds.. SIMUNEK J. H.388 LÖSCHER W. Plasma concentrations of chloramphenicol in birds.de. et al.. SCHERKL R.Without doubt the mode of application by capsule or tablet is . KRAUTWALD-JUNGHANNS. KAMPHAUSEN L. THOMAS JE. KROKER R. 6. et al. LÖSCHER W. UNGEMACH F.. Berlin: Parey Buchverlag 1999: 230 . 04103 Leipzig. Nitrofurane. DiplECAMS Clinic for Birds and Reptiles. Pharmakotherapie bei Hausund Nutztieren“.): Pharmakotherapie bei Nutz. CLARK CH. 43: 1249 – 1253. In: LÖSCHER. E. Auflage.231 AUTHORS ADDRESS M. GYLSTORFF I AND GRIMM F. An den Tierkliniken 17. Am J Vet Res 1982. Nitrofurane. The disadvantages of the application by capsule are that it is time consuming and impracticable for the treatment of a larger collection (KAMPHAUSEN 2003).und Haustieren. MILTON JT. (eds. regurgitation could be seen after capsule application in 3 pigeons. The results indicate that in African grey parrots. despite the observed immune-depression in young birds of this species. all 29 were tested PBFDV-positive by PCR with a positive PBFD-signal predominantly in skin-feather-samples (n=26). H. 1989). Considering the high risk of contaminated PCRsamples from the skin and/or feathers by dust. a member of the genus Circovirus. the psittacine beak and feather disease virus (PBFDV) has been reported in some other bird species (RAHAUS and WOLFF 2003. Whereas in 21 birds viral DNA was detected in more than one sample (skin 18/21. in eight birds only the skin-feather sample was PBFDVpositive. we screened skin-feather samples and primary lymphoid tissues (spleen. pigeon circovirus. infection with the aetiological agent. PBFDV is. Köhler. caused by psittacine beak and feather disease virus (PBFDV).Acute outbreak . goose circovirus and canary circovirus should also be grouped in this genus (MANKERTZ 249 . spleen (1/3) and bursa (2/3). R. bursa 9/19). Korbel KEYWORDS PBFD . Nonetheless. biopsy specimen from the liver seems to be an alternative for individual diagnostic of PBFDV in African grey parrots. as well as liver. the liver is the primary target organ. To detect diseased birds or virus carriers. In addition. B. bursa of Fabricius) in parallel. Ludwig-Maximilian-Universität München.Diagnostic . Grund. EISENBERG et al.Klinik für Vögel. Germany EVALUATION OF VARIOUS TISSUES FOR DIAGNOSIS OF PSITTACINE BEAK AND FEATHER DISEASE (PBFD) C. 1 INTRODUCTION Beak and feather disease (PBFD) is a severe disease in most species of the order Psittaciformes (RITCHIE 1995). spleen 9/21. which suffered an outbreak of acute PBFD. T. 2003). Of a total of 34 birds investigated.PCR . By homologies of the viral genomes.African grey parrots ABSTRACT Psittacine beak and feather disease (PBFD) is an important viral disease of psittacine birds. In order to evaluate the reliability of PCR results obtained by testing skin-feather samples. in birds tested PBFDV-negative in the skin. like the porcine circovirus. liver 21/21. PCR is the diagnostic method of choice. viral DNA could be detected in liver (3/3). Samples were derived from a flock of young African grey parrots (Psittacus erithacus). of the family Circoviridae (RITCHIE et al. Germany). 20s 57°C. 2 MATERIAL AND METHODS 2. 250 . In particular.2. 2000. blood and feathers are used routinely for diagnostic PBFDVPCR.2mM dNTP`s. which may result in lethal secondary infections and sometimes accompanied by an irreversible loss of feathers. The 717bp long PCR products were separated in a 2% agarose gel and stained with ethidium bromide. Considering. 2. that virus load might vary considerably in different species depending on the course of infection we investigated tissue distribution in 34 African grey parrots that died during an outbreak of acute PBFDV infection by PCR. DNA extraction DNA extraction was done by a commercial DNA-isolation kit (DNA easy tissue kit.et al.2 IU of taq-Polymerase (peqlab. in order to obtain reliable results by this fast and sensitive method. 2. However. The thermocycler conditions were: 15 min 95°C (activation of the polymerase and initial denaturation) followed by 32 cycles of 30s 95°C. 30s 72°C and a final extension at 72°C for 5 min. 0. in African Grey parrots this acute course of PBFDV infection has been recognised in young birds that died with no obvious symptoms (SHOEMAKER et al. TODD et al. Polymerase chain reaction Isolated DNA was tested for the presence of circoviral DNA by PCR amplifying a part of the genomic region within ORF 1 applying the protocol (primer no. Erlangen. independent of the species tested. Diagnosis of PBFDV infection relies most often on detection of viral DNA by polymerase chain reaction (PCR). Samples taken for PCR-test included the spleen. Hilden. primary lymphoid organs (spleen and bursa of Fabricius) as well as liver samples in parallel. (1999) with some modifications. Quiagen. the other organs with two injection needles. Characteristic signs common in chronic cases. Animals and experimental design From a flock of African grey parrots (Psittacus erithacus) that suffered an acute outbreak of PBFD. 34 birds were sent in for pathological examination. Germany) according to the manufacturer’s instructions.1. we used 25µl reaction mixture containing 25pmol of each primer. selection of the sample is crucial. The birds were delivered together in big plastic bags in frozen condition. Testing skin-feather samples. are a slow progressive loss of feathers in conjunction with beak elongation and necrosis of beak and palate. 2 and 4) described by YPELAAR et al. reliable diagnostic samples for PBFDV-PCR in African grey parrots should be identified.3. bursa of Fabricius and skin with feathers. 2000). 1mM MgCl2 and 0. paying attention to change instruments between each sample. liver. the course of the disease is dominated by an acute onset of a severe immunosuppression. In young birds however. 2001). Skin with feather samples was cut with fresh razor blades. All samples were stored at -70°C until further processing. clinical outcome of infection may vary. In brief. So far. Depending on the species and age of birds. 251 . 1: PCR-results of birds from an acute PBFD-outbreak. a total of 34 African grey parrots from a suspected outbreak of acute PBFD were investigated for gross anatomic changes as well as for viral infection by PCR. In eight birds a discreet red discolouration of the contour feathers in the dorsal and ventral caudal abdominal region was visible. In contrast a positive PBFDV-signal in bursa and spleen was obtained only randomly (spleen: 9/31. Viral DNA was predominantly detected in samples of the skin and feathers (26/32) and liver (21/33). bursa: 9/27). as well as a hydropericardium (n=11). Predominant alterations of internal organs included hepatomegaly (n=15) and splenomegaly (n=12). viral DNA could be detected in liver (3/3). spleen (1/3) and bursa (2/3) (Fig 1B). However. Fig. bursa n=7) were tested PBFDV-positive (Fig. in 3 birds tested PBFDVnegative in the skin. Testing for PBFDV by PCR a total of 29 birds were tested PBFDV-positive.3 RESULTS For this study. 21 had a positive PCR-signal in two or more tissues. 1A). Overall. Within the group of PBFDV-positive birds (n=29). the birds` body condition was good except for seven birds with a light to moderately reduced bodyweight. At external inspection birds had complete plumage with normal formed feathers and the horn of beak and claws was intact. Investigating the birds where PBFDV was detected in the skin-feather-samples (n=26) only three quarter of the liver-samples (n=18) and one third of the lymphoid organs (spleen n=8. with the respective percentage given above. 1C. the high frequency of PBFDV in the liver emphasise the relevance of the liver for pathogenesis of acute PBFD in young African grey parrots. In both groups viral DNA was detected also in the liver. in the group of PBFD-positive birds with negative PCR-results of the liver-samples (Fig 1D. In addition. 2000).From 34 African grey parrots. in a longitudinal study of PBFDV infected budgerigars (Melopsittacus undulatus) viral DNA could be detected in feathers of birds without clinical signs for as long as 38 weeks (HESS 2004). 29 birds were tested positive for PBFDV by PCR. E PBFDV-positive spleen sample (n=9) and F PBFDV-positive bursa sample (n=9). PBFDV-positive samples. This frequent detection of PBFDV in the liver is in agreement with observations by Shoemaker et al. that viral inclusion bodies were detected regularly in the bursa (7/7). bursa 9/19). biopsy specimen from the liver for PCR seems to be an alternative for the diagnosis of PBFDV in individual birds in this species. n=9) or bursa of Fabricius (Fig 1F. but not always in the skin-feather samples. that feathers harvest virus. 4 DISCUSSION In the present study of 34 African grey parrots from a flock that suffered an outbreak of acute PBFD. in particular a positive PCR-signal only in a skin-feather sample. C PBFDV-positive liver sample (n=21). In contrast. he was able to demonstrate moderate to severe coagulative necrosis in the liver. Nonetheless. that within the group tested positive in the liver (Fig. In addition. (SHOEMAKER et al. n=21) almost 1/3 of the lymphoid organs (spleen n=9. it became evident. D PBFDV-negative liver sample (n=7). Considering that acute PBFD is seldom associated with feather manifestations (SHOEMAKER et al. In our study we were able to demonstrate viral DNA in the bursa as well as in the spleen. In particular. B PBFDV-negative skin/feather sample (n=3). PCR-samples from the skin and/or feathers have a high risk of contamination. Considering that PBFDV can spread by feather dust. In a study on 14 young African grey parrots. Comparing the PCR-results of the internal organs. 252 . Columns in blue represent the number of samples tested and in red. In addition to skin-feather samples (n=26) viral DNA was detected regularly in the liver (n=21). it is known. even without alterations. but not as often as in the liver (spleen 9/21. suffering acute PBFD. Concerning our results in African grey parrots. However. the lymphoid tissues (bursa and spleen) gave also negative results (Fig 1D). Similar results were observed when PBFDV-DNA was detected in the spleen (Fig 1E. bursa n=9) were tested PBFDV-positive as well as 18 skin-feather samples. n=9). n=7). For example. the regular detection of PBFDV-DNA in skin-feather samples is somewhat surprising. Shown are the PCR-results of these 29 birds divided in groups with A PBFDVpositive skin/feather sample (n=26). the comparison of the PCR results from spleen and liver proves that the birds did not die during viremia and thus the positive PCR-signal indicates replication within the liver. 29 birds were tested PBFDV-positive by PCR. he reported. as observed in 7 birds in this study has to evaluated critically. 2000). 4. 171(1): 83 .88. Vet Microbiol 1999. Avian Pathol 2004.371. J Virol 2001. Lake Worth: Wingers Publishing 1995: 223 .uni-muenchen. 9. Beak and feather disease virus and porcine circovirus genomes: intermediates between the geminiviruses and plant circoviruses. 85764 Oberschleißheim. MANKERTZ A. 286.5 CITATION INDEX 1. 44(2): 470 . Detection of circovirus with a polymerase chain reaction in the ostrich (Struthio camelus) on a farm in the Netherlands. Email: Christian. Grund. Avian Dis 2000. RITCHIE BW. 354 . a new circovirus from pigeons. AUTHORS ADRESS C.2479. YPELAAR IA.38. 145: 2469 .Grund@lrz. RAHAUS M. H. Cloning and sequencing of columbic circovirus (CoCV). 95: 27 . Characterization of a new virus from cockatoos with psittacine beak and feather disease.W. SCHOEMAKER NJ. In B.478. 3.1744. Sonnenstrasse 18. Universal polymerase chain reaction for the detection of psittacine beak and feather disease virus. Arch Virol 2000. Circoviridae.362. EISENBERG SWF. Ludwig Maximilian Universität. cloacal swabs and blood from budgerigars (Melopsittacus undulatus.de 253 . Comparative sensitivity of polymerase chain reaction diagnosis of psittacine beak and feather disease on feather samples. RITCHIE BW. Severe leukopenia and liver necrosis in young African grey parrots (Psittacus erithacus erithacus) infected with psittacine circovirus. 10. Vet Microbiol 2003. 33: 477 . 2. HESS M. 6. 68: 141 148. Shaw 1805). Psittacine beak and feather disease: a first survey of the distribution of beak and feather disease virus inside the population of captive psittacine birds in Germany. TODD D. 143 (9): 1723. Virology 1989. Arch Virol 1998.).252. 7. Genome sequence determination and analyses of novel circoviruses from goose and pigeon. J Vet Med B 2003.481. NIAGRO FD. 5. Klinik für Vögel. Avian Viruses: Function and Control. 50: 368 . 8. RITCHIE (Ed. Bet Dagan. We conclude that the crop should not be recommended for clinical PDD diagnosis due to the high prevalence of false-negative cases.Crop biopsy ABSTRACT A pair of blue and yellow macaws (Ara ararauna) was introduced into a breeding farm in Israel and produced chicks that presented with proventricular dilatation disease (PDD) signs at the age of 10-14 wks. The rest of the birds did not present any histologic changes. Israel RELIABILITY OF CROP BIOPSY AS A DIAGNOSTIC TOOL FOR PROVENTRICULAR DILATATION DISEASE IN PSITTACINE BIRDS U. A. Bendheim1. Nor does a histopathological 254 . Edery2. while only 4 birds had similar microscopic changes in the crop. 19 of the euthanased birds were necropsied for comparison of crop vs. PhD. Kimron Veterinary Institute2. DVM. regurgitation and indigestion. as central and peripheral nervous tissues are involved. Dip ECAMS. 1 INTRODUCTION Proventricular dilatation disease (PDD) was first described in macaws as a malabsorption/maldigestion disorder. Birds intended for breeding were euthanased. Clinically PDD is not easy to confirm in the live bird. and were processed for histological examination. Square samples of 1 cm2 including blood vessels and probably nerves were taken from these tissues. Forty two grey parrots (Psittacus erithacus) and 8 blue and yellow Macaws were hatched in this aviary in that period. but central nervous signs may also be observed. DVM KEYWORDS Proventricular Dilatation Disease (PDD) . proventricular and brain lesions to test whether crop biopsy may be a reliable means for the clinical diagnosis of PDD. Lublin 2 DVM. Hebrew University. N. about 27 of whom died and demonstrated lesions typical of PDD. Common clinical signs are weight loss. because x-ray imaging does not give a clear diagnosis (GREGORY 1995) and because of the lack of serology.Koret School of Veterinary Medicine1. Also brain and proventriculus may present negative histological results even in birds with PDD. Jerusalem. Histology demonstrated multifocal lymphohistiocytic infiltration with gliosis in brain and infiltration in ganglia of the muscularis externa of the proventriculus in 10 birds out of 19. 33% of birds with lymphoplasmacytic proliferation in the proventriculus did not have proventricular dilatation (GREGORY 1995). all birds presented dilatation of the proventriculus. refusal to eat pelleted food. and one chick had blood in the faeces and in the regurgitation content.5 40 75 Total 69 * Recovered after treatment. and were left for follow up. about one year after the introduction of a blue and yellow macaw (Ara ararauna) breeding pair to the farm. Most of the chicks became sick at 10-14 wks. In post mortem. typical lesions were found in the proventriculus of all birds. Sporadic cases of PDD have been observed in adult psittacine birds (various species) in Israel in the past. following the veterinarian’s advice. agreed to euthanase the clinically affected birds intended for breeding. In a severe outbreak in a psittacine breeding farm with high mortality which was diagnosed as PDD the owners. The three remaining chicks were treated for 1 month with 10 mg per day of celecoxib (CELCOX 100). GERLACH (1991) found microscopic lesions in only 62% of birds with typical PDD clinical signs. towards weaning time the chicks refused to eat spontaneously and even refused to accept the usual amount of forcefeeding. One bird revealed severe respiratory distress. but have not been published. with histological confirmation of PDD in 12 out of 19 cases (63%). growth retardation and in most cases also regurgitation.evaluation of suspected cases give a clear diagnosis. which belongs to a new family of non-steroidal antiinflammatory drugs (NSAIDs) (DAHLHAUSEN and ALDRED 2004). However. 255 . Distribution of morbidity and mortality of the various species is presented in Table 1. In a group of 15 psittacine birds with proventricular dilatation. Species Grey parrot Blue and yellow macaw Citron-crested cockatoo Galah Eclectus N 42 8 6 8 5 Healthy 3* 0 4 7 3 17 Sick birds euthanased 17 1 0 0 0 18 Died 22 7 2 1 2 34 %Affected (morbidity & mortality) 93 100 33 12. In most cases the disease lasted for 2-10 days before death. Distribution of morbidity and mortality of various species is a breeding farm in which PDD occurred. Table 1. 2 CASE HISTORY A severe outbreak of PDD occurred in year 2004 in chicks of a psittacine breeding farm in Israel. All sick chicks except 3 were euthanased. in order to prevent further spreading of the disease. Three of the chicks also had central nervous signs (torticollis). one chick at 30 wks. The clinical signs were: stasis of the crop. but only in 67% in the crop (GREGORY 1995). Signs of clinical PDD 19 PDD in gross pathology 19 Histological changes in brain 10 Histological changes in proventriculus 10 Histological changes in crop 4 Differential diagnosis: To eliminate other diseases that may be involved in nervous signs. Candida albicans was isolated. Proc Euro Assoc Avian Vet. together with a brain sample. and were stained with hematoxylin and eosin. Lake Worth: Wingers Pub 1995.il 256 . proventriculus or crop (at least in one of these organs). CITATION INDEX 1. These included: Indian ringneck parakeet (n=20). All premises in which the affected birds were kept were disinfected by formaldehyde evaporation. Resolution of clinical proventricular dilatation disease by cyclo-oxigenase 2 inhibition. 3 PATHOLOGY Nineteen of the dead birds were necropsied (14 grey parrots. bacterial and mycotic agents in brain. analysis of species.net. DIP ECAMS P.t AUTHORS ADDRESS U. In addition.Box 196. 3. the rest results were all negative. DAHLHAUSEN B and ALDRED S.81. sun conure (n=3). Israel Email: dr-uri@012. black-headed caique (n=5). (2004).O.Several species remained free of any clinical signs. Shavey Zion 25227. tests for Newcastle disease. segments of 1 cm2 were cut from crop and proventriculus and were fixed in a 10% formaldehyde solution. The typical microscopic lesions were multifocal lymphohistiocytic infiltration with gliosis in brain and lymphohistiocytic infiltration in ganglia of the muscularis externa of the proventriculus and/or the crop. and Chlamydophila psittaci (serology and immunofluorescence) were carried. West Nile disease (serology). Vienna 1991. epizootiology. From these birds. GERLACH H. diagnosis and virological results. Macaw wasting disease. Pionus spp (n=6). The macaws that had been suspected as the origin of this outbreak of PDD were transferred to a non-breeding aviary. 3 blue and yellow macaws and 2 citron-crested cockatoos). 444 .445. Table 2. DVM. GREGORY CR. The diagnosis of PDD was confirmed in 12 of the birds following a histopathological examination of the brain. Proventricular dilatation disease. 2. Summary of histological changes in 19 birds with clinical PDD. 273 . The histological changes in the examined birds are summarized in Table 2. the crops were examined for fungi. Bendheim. and in 3 of them. Dilatation of the proventriculus was observed in all necropsied birds. A 4 year study on clinical case history. In: RITCHIE BW (ed): Avian Viruses Function And Control. avian influenza. and lory (n=2). Jardine’s parrot (n=6). weight loss.Passeriformes .Myenteric ganglioneuritis . The most common clinical signs of PDD include depression. Universidad Autonoma de Barcelona. Barcelona. is described in 4 species of non-psittacine birds (3 of the order Passeriformes and 1 of the order Piciformes). LV. Parque Zoologico de Barcelona2. 1998). and characterized histologically by lymphoplasmacitic infiltrates of the myenteric plexus of the gastrointestinal tract. H. Maragall ExoticsVeterinary Centre3. a chaffinch Fringilla coelebs. 2003). especially the nerves that supply the muscles in the proventriculus and other digestive organs including crop. LV. passage of indigested seeds on the faeces and/or central nervous system signs (RITCHIE et al. Barcelona. Canada geese and rosette spoonbills (RITCHIE et al. weaver finches. canaries. Ramis. DAOUST et al. regurgitation. and an Amazon umbrella bird Cephalopterus ornatus) and 1 species of Piciformes (a bearded barbet Lybius dubius). This paper describes 4 non-related cases of a PDD-like syndrome in 3 species of Passeriformes (a canary Serinus canaria. Barcelona. Although this syndrome is typical of psittacine birds. honey-creepers. C. 1 INTRODUCTION Proventricular dilatation disease (PDD) is a condition of psittacine birds characterised histologically by an inflammatory response with accumulation of lymphocytes and plasma cells in the nervous system. red-tailed hawks. LV. and splachnic neuropathy (SCHMIDT et al. MS. LV. suggestive lesions compatible with PDD have also been reported in toucans. 1998. A. The disease has also been called neuropathic gastric dilatation. MS. PhD. Perpiñán1. Dipl ECVP KEYWORDS Birds .Piciformes . 2003). 257 .Proventricular dilatation disease ABSTRACT A clinical syndrome similar to proventricular dilatation disease of psittacines.Veterinary practitioner1. López3. Fernández-Bellon2. Spain MYENTERIC GANGLIONEURITIS IN FOUR NON-PSITTACINE BIRDS D. 1998). 1991. myenteric ganglioneuritis. Facultad de Veterinaria. Departamento de Medicina y Cirugia3. SCHMIDT et al. ventriculus and small intestine (RITCHIE et al. 517. the bearded barbet and the Amazon umbrella bird. In the present paper we describe 4 non-related cases that clinically. this condition must be suspected in birds with wasting disease or crop problems. 2000). Gross necropsy findings were variable and included emaciation. the disease seemed to be limited to macaws. The description of myenteric ganglioneuritis (or PDD-like syndrome) in multiple families of birds may suggest that its cause is not restricted to a particular host (GREGORY et al. hepatic and renal atrophy or enlargement. YASON CV and ARTSOB H. GREGORY et al. 27(3): 513 . Histologically. weakness. all 4 cases presented multifocal lymphoplasmacytic myenteric ganglioneuritis. 2000). 2002) and their use should be also considered in non-psittacine birds. since then PDD has been reported in more than 50 species of Psittaciformes (GREGORY et al. and the characteristic myenteric ganglioneuritis. Initially. The bearded barbet was found dead in its enclosure. SCHMIDT et al. E. Three of the species affected (canary. DAOUST P-Y. varied for each case. However. 2. 3 DISCUSSION Proventricular dilatation disease (PDD) was first described in the late 1970s. DALHAUSEN B. macroscopically. Although suggestive lesions of PDD in some non-psittacine species have also been orally communicated (GREGORY et al. 4 CITATION INDEX 1. Although myenteric ganglioneuritis can show a range of clinical signs. were housed at the Barcelona Zoo. J Wildl Dis 1991.2 CLINICAL CASES Two of the birds.12. chaffinch and Amazonian umbrella bird) were Passeriformes and the fourth (bearded barbet) was a member of the family Piciformes. Proventricular impaction associated with nonsuppurative encephalomyelitis and ganglioneuritis in two Canada geese. and intestinal and ventricular dilatation. 258 . gallbladder dilatation. Monterey 2002: 9 . Although specific treatment was not attempted in the present cases. since they combine gastrointestinal signs and lesions. ALDRED S and COLAIZZI. Spain. Cardiac lymphoplasmacytic ganglioneuritis was also observed in all cases. cyclooxigenase-2 inhibitors have been used successfully in psittacine birds with PDD (DALHAUSEN et al. JULIAN RJ. Resolution of clinical proventricular dilatation disease by cyclooxigenase 2 inhibition. there is only one published article that reports a condition similar to PDD in 2 Canada geese (DAOUST et al. The intensity of the lesion. Proc Assoc Avian Vet. and/or ataxia. including polyphagia. and the canary and the chaffinch were kept as pets and were presented to the Maragall Exotics Veterinary Centre in Barcelona. and microscopically are similar to psittacine PDD. as well as the degree of involvement of different GI sections. especially when the results of crop cytology are unrewarding. and the other animals showed varied clinical signs before their death. 2003). without previous signs. 2000. weight loss. 1994. 1991). 75. AUTHORS ADDRESS David Perpiñán C/ Balmes. Polyomavirus. RITCHIE BW. St Paul 1998: 25 . J Assoc Avian Vet 1994. et al.40. GREGORY CR.65. GREGORY CR.3. 17. Proc Assoc Avian Vet. 8(2): 69 . Progress in understanding proventricular dilatation disease. Gastrointestinal system and pancreas. Proc Assoc Avian Vet. and PBFD virus.275.es 259 . Spain Email: dperpinan@yahoo. et al. SCHMIDT RE. In: Pathology of pet and aviary birds 2003. REAVILL DR and PHALEN DN. 08918 Badalona (Barcelona). LATIMER KS. GREGORY CR. NIAGRO FD. Iowa: Iowa State Press: 41 . 6. RITCHIE BW. Portland 2000: 269 . et al. LATIMER KS. LATIMER KS. A review of proventricular dilatation syndrome. 5. 4. Progress in preventing PDD. Test . increase of fungal exposure. Deville1. depression. psittacine and waterfowl. M. 38 avian sera were considered “positive” for antigenemia analysis. For all these birds aspergillosis was suspected on the basis of clinical signs and/or radiographical examination. was detected by counter-immunoelectrophoresis. Centre de Sauvegarde2. France EVALUATION OF THE SEROLOGICAL TEST PLATELIA® ASPERGILLUS FOR THE DIAGNOSIS OF ASPERGILLOSIS G.Platelia . In 14 out of these sera. the presence of precipitating antibodies to Aspergillus spp. E. especially in psittacine. Usually the disease does not occur when the bird is healthy and exposed to a low concentration of fungal conidia. under certain circumstances (e. especially in birds of prey. J. cyanosis or sudden death without any clinical signs. Ecole Nationale Vétérinaire d’Alfort. We conducted a prospective study during the years 2003 and 2004. This test is based on the detection of fungal antigen (galactomannan) by sandwich ELISA. raptors and aquatic birds in captivity. Maisons-Alfort.Service de Parasitologie-Mycologie1. Risi2. Guillot1 KEYWORDS Serology .g. Faculté de Médicine de Créteil. Using the Platelia® Aspergillus kit. Ecole Nationale Vétérinaire de Nantes3. Service de Pasitologie.Aspergillus fumigatus . Avian aspergillosis usually presents fairly nonspecifically with inappetance. Bretagne3.Serological . This study clearly indicated that the detection of Aspergillus antigens should not be considered as a valuable test for the diagnosis of aspergillosis in birds. 1 INTRODUCTION Aspergillosis is a common and life threatening disease in wild and domestic birds. S.Aspergillosis ABSTRACT Aspergillosis remains a major cause of mortality in birds. UMR BIPAR. Le Loch’1. On the other hand. The condition is caused by saprophyte fungi of the genus Aspergillus. A total number of 141 serum samples from 33 different species were collected from privately-owned psittacines or wild birds at two wildlife refuges in France: Nantes and Alfort. weakened immunity) 260 . dyspnoea. The objective of the present study was to evaluate the accuracy of the serological test Platelia® Aspergillus for diagnosis and follow up of cases of aspergillosis in different avian species. with Aspergillus fumigatus being the predominant species involved. is the antigen whose detection by ELISA (Platelia® Aspergillus) allows diagnosis of invasive aspergillosis in humans. or because the results of complementary tests suggested the presence of aspergillosis. Sometimes. combined with radiography. The serum was separated by centrifugation at 1000g for five minutes. History and clinical examination of the bird. provide clues to the presence of lesions due to Aspergillus. OGLESBEE 1997. We recently conducted a feasibility study on birds to assess the efficiency of Platelia® Aspergillus for the diagnosis of aspergillosis. etc. The sera were collected either because birds showed clinical signs of aspergillosis.Aspergillus can develop in the body and lead to non-specific clinical signs. After electrosyneresis. respectively) and that specificity is the same (93%) (LATGE et al. the sera were frozen at -20°C 261 . KAWAZU et al. An indirect ELISA has been developed specifically for birds in raptor centers.g.5ml Eppendorf tubes. MENNINK-KERSTEN et al. histology and culture. but these tests require the anaesthesia of the bird. MENNINKKERSTEN et al. This ELISA is not available in France. The treatment of aspergillosis is difficult and often not efficient because the diagnosis is made too late. The diagnosis of avian aspergillosis remains a challenge for veterinarians. Serological testing for the anti-Aspergillus antibodies assay is used and usually allows an efficient and early diagnosis. 2 MATERIALS AND METHODS One hundred and twenty-three sera collected from 116 birds were sent to the Parasitology department at the veterinary college of Alfort (ENVA) by veterinarians or wildlife care centers. a polysaccharide composing the cell wall of Aspergillus fumigatus. endoscopy and a complete blood count. 2004. REDIG 1993). Some serum samples were also collected because the bird’s situation rendered it vulnerable to aspergillosis: e. Moreover. In human medicine. aspergillosis is a major disease in haematology and oncology care units. GRACZYK et al. the bird dies before any clinical signs appear (BAUCK 1994. Galactomannan (figure 1). OROSZ 2000). Diagnosis must be made early to allow efficient treatment. birds exposed to excessive stress. 1998. Clinical signs are diverse and often late in becoming evident. LATGE et al. 1994. the sera were conserved at 4°C until the moment they underwent electrosyneresis (a few days). The detection of Aspergillus is achieved by cytology. 2004). 2004). young birds recently purchased. JONES and OROSZ 2000. and the necessary handling can threaten the life of a bird under respiratory distress (KAWADU et al. studies for the detection of Aspergillus DNA have been made and are promising (PHALEN 2001. Stored in 1. It has recently been shown that the use of serological testing could resolve this problem. where veterinarians use electrosyneresis to detect precipitant antibodies. 2004. wild birds held in wildlife centers. 1994. A comparative study in humans has showed that the sensitivity of ELISA Platelia® Aspergillus is higher than the sensitivity of a PCR test (test GeniQ-Asper®) (100% and 55%. EBA-2. The Platelia® Aspergillus test is an immuno-enzymatic assay directly on a solid phase. which allows the semi-quantitative detection of galactomannan circulating in the serum. This is a monoclonal antibody.. A serum sample is considered to be positive when it forms at least one clear precipitation arc with at least one of the two types of antigens.up to the moment of undergoing the ELISA test. on which they migrated towards the anode and cathode. when marked with peroxidase. 1994) galactomannan galactofurans. The number and intensity of the precipitation arcs varied as a function of the concentration of precipitant antibodies. KAWAZU et al. GRACZYK et al. called an ELISA sandwich. LATGE et al. and. visible after staining. 30µl of each antigen and 15µl of sera to be tested were placed in the presence of an electric field on cellulose acetate strips. This test has a positive threshold fixed at 1ng/ ml of serum tested. which recognizes the � (BAUCK 1994. 1998). 2004. The first step in the reaction consists of treating the sera in order to free the circulating immuno- 262 . JONES and OROSZ 2000. 2004) For electrosyneresis. it is used as a conjugate for the detection of antigens. Contact between antigens and antibodies leads to the formation of one or several precipitation arcs. but considering the small volumes of the sera we collected. we used only 100µl. as well as a test to detect galactomannan (Platelia® Aspergillus test). These instructions advise using 300µl of sera. An IgM antibody was used. The positive control serum came from a dog in which aspergillosis was confirmed. Each serum sample was subject of a serological test to detect anti-Aspergillus antibodies (electrosyneresis). 1998. The ELISA test was conducted using the Platelia® Aspergillus Bio-Rad kit instructions (GRACZYK et al. It has a double use because it is fixed at the bottom of the microtitre wells. respectively. Two types of antigens were used in the electrosyneresis: metabolic and somatic Aspergillus fumigatus antigens (Bio-Rad). Fig 1: structure of galactomannan (MENNINK-KERSTEN et al. two threshold sera (1 ng/ml of galactomannan) and one positive control serum (10ng/ml of galactomannan). For each well. The peroxidase enzymatic activity was revealed by the addition of 200µl of dimethylsulfoxide (DMSO) in each well. the birds showed no clinical signs.000g. 100µl of each serum is vigorously mixed with 33µl of an EDTA acid solution (ethylene dinitrolo tetra acetic acid). the plates were washed five times with a TRIS-NaCl buffer solution. and 7 species belonging to various taxonomic families. The support plate was then incubated at 37°C for 90 minutes. Once the sera were prepared.e. an index (I) was calculated for each serum tested as follows: I = OD sample / TV The aim of this calculation is to limit the variations in the inter-trial and inter-laboratory OD.complexes and to precipitate the proteins that can interfere during the immunoenzymatic reaction. with the majority being Psittacidae (17 species). They consist of one negative control serum. The test is validated if the following criteria are respected: 0. 263 . and 38 birds older than one year. The reaction was ended after 30 minutes by the adding of 100µl of a sulfuric acid stopping solution to the wells. 6 raptor species. complexes of antibodyantigen-antibody + peroxidase form. 38 were sampled by simple screening i.8 I (R5) > 2.0 ≤ I < 1. The readings were taken by a spectrophotometer at a wavelength of 620nm. Following incubation.5 The results can then be interpreted as follows: I < 1. Thirty species were represented. Of those birds for which an age could be determined. In the presence of antigens. To achieve this. there were 39 birds aged one year or less.5: serum “intermediate” for the presence of galactomannan I ≥ 1. On determining a threshold value (TV) corresponding to the average OD of the wells containing the threshold serum (R4).3 ≤ TV ≤ 0. then with 50µl of supernatant for each serum treated.0 I (R3) < 0.0: serum “negative” for the presence of galactomannan 1.5: serum “positive” for the presence of galactomannan 3 RESULTS AND DISCUSSION Among the 116 birds from which serum samples was collected. heated at 100°C for three minutes. the microtitre wells (in which the EBA-2 antibodies are fixed) were filled with 50µl of conjugate (EBA-2 antibodies coupled with peroxidase). and then centrifuged for 10 min at 10. an optical density (OD) was determined. Four control sera supplied with the kit were prepared in the same manner. We can suppose that the immune system.5 > I 4 ≥ 5.Sula bassana). and Group C (aspergillosis considered as unlikely) comprised birds showing a negative electrosyneresis result and no clinical signs. immuno-suppression becomes very important and the circulating level of galactomannan rises sharply.5 > I 2 ≥ 3. when galactomannan is present in a bird serum. As a consequence.5 ≥ 1>I 0. we were able to calculate the sensitivity and specificity of the Platelia® Aspergillus test: 30% and 86%.5 and 3. 55 50 45 40 Number of sera 35 30 25 20 15 10 5 0 I < 0.5 were in poor general condition. Group B (presumptive aspergillosis) comprised birds showing only one of the two characteristics shown by group A.Of the 123 serum samples tested. only 18 per cent of the birds whose index was between 1.5 ≥ 5>I 4. But as soon as it is surpassed. respectively.5 Index Figure 2 : distribution of sera according to their index According to clinical signs and electrosyneresis results the birds were categorized into three groups: Group A (aspergillosis considered as highly probable) is made up of birds giving a positive electrosyneresis result and showing symptoms compatible with aspergillosis. insofar as it remains competent.5 (15. Fifty-six per cent of the birds whose index was higher than 3.5 ≥ 3>I 2.5 > I 3 ≥ 4. 35 gave a positive Platelia® Aspergillus result (figure 2). Conversely.2ng/ml of galactomannan) and 18 of these sera had an index above 3.2ng/ml of galactomannan). it is present in high concentrations. The average index of the 35 positive serum samples was 3. Given the low sensitivity of the test in 264 ≥ 6>I 5.5 > I 5 ≥ 6.5 ≥ 1.5 ≥ 4>I 3. allows the bird to maintain a relatively low level of antigens.5 > I 6 ≥ I6.39 (approximately 11.5 ≥ 2>I 1. and for at least one of these birds (a gannet .5 were in poor general condition.5 . Considering birds of group A as infected and birds of group C as healthy.5 > I 1 ≥ 2. aspergillosis was confirmed by culture and isolation of Aspergillus fumigatus from a pulmonary granuloma. Sem Avian Exot Pet Med 2000. The Platelia® Aspergillus test is capable of detecting galactomannan at a concentration of 1ng/ml. NANNYA Y.2741. Mycoses. Value of antigen and antibody detection. appears to be limited. which can give a positive result on the ELISA test.the present study. when used alone. An investigation with confirmed cases of invasive aspergillosis would be necessary to confirm the usefulness and the performance of the Platelia® Aspergillus test in these specific cases. In humans. Some of these fungi are occasionally found in birds. 2733 . only a few false positive results occurred with the Platelia® Aspergillus test and the specificity (86%) and the positive predictive value (75%) of the test were high. double-sandwich enzyme-linked immunosorbent assay for galactomannan. moulds of the genera Penicillium. false positive results have been explained by positive reactions of ELISA with the lipoteic acid in Bifidobacterium. It would be interesting to know if these birds develop aspergillosis later on. Prospective comparison of the diagnostic potential of real-time PCR. CRANFIELD MR and KLEIN PN. Finally. 3. OGLESBEE 1997). Mycopathologia. KANDA Y. The Platelia® Aspergillus test is thus relevant for the detection of circulating galactomannan. Furthermore. GRACZYK TK. The diagnostic of aspergillosis in birds.58. without any signs of the infection) showed a positive result for the Platelia® Aspergillus test. since eight birds out of the 38 tested by random screening (i. but not for the diagnosis of avian aspergillosis. and a (1�3)-�-D-Glucan test in weekly screening for invasive aspergillosis in patients with hematological disorders. 4 CITATION INDEX 1. KAWAZU M. 9(2).1006.127. Alternaria. BAUCK L. Lake Worth: Wingers Publishing 1994. On the other hand. 2. The Platelia® test could be useful in the detection of avian aspergillosis before the onset of clinical signs. It thus seems that the circulating concentrations of galactomannan in birds infected by Aspergillus are frequently lower than 1ng/ml. 52 . 1998. 140. This could be explained partly by the low vascularisation of certain infected organs such as the air sacs. 42. et al. In: RITCHIE BW. J Clin Micro 2004. 997 . the usefulness of this test for diagnosing aspergillosis in birds. and blood evaluation parameters in diagnosis of avian invasive aspergillosis. and Paecilomyces all contain galactomannan. 121 . HARRISON GJ and HARRISON LR (eds): Avian Medicine: Principles and Application. We may thus presume that the Platelia® Aspergillus test is better advised for the diagnosis of invasive aspergillosis than for localized forms of the infection.e. we observed that birds showing signs of invasive aspergillosis yielded a positive result for the Platelia® Aspergillus test more frequently (45%) than those showing only signs of respiratory aspergillosis (23%). 4. 265 . as well as the time between galactomannan detection and the appearance of the first clinical signs. Recent study demonstrated that a low antigenemia level may be common in healthy birds (JONES and OROSZ 2000. JONES MP and OROSZ SE. 327. CLUBB SL. LATGE JP. Sem Avian Exot Pet Med 2001.5. Sem Avian Exot Pet Med 2000. Ecole Nationale Vétérinaire d’Alfort.89. et al. (eds): Avian medicine and surgery. The Lancet 2004. 8. MENNINK-KERSTEN M. In: FOWLER ME (ed): Zoo and Wild Animal Medicine Current Therapy. France Email: guillaume_ll@hotmail. 59 . 6. DORRESTEIN GM. PHALEN DN. Philadelphia: WB Saunders 1997. 323 .181. Mycotic diseases. 5424 . 7. 9. UMR BIPAR. WARRIS A. KOBAYASHI H. 77 . 325 . Infection and Immunity 1994. OROSZ SE. Overview of aspergillosis: pathogenesis and treatment options. DEBEAUPUIS JP. 9(2).com 266 . The use of serologic assays in avian medicine. In: ALTMAN RB. 10(2). 178 . KLONT R. Philadelphia: WB Saunders 1993. Chemical and immunological characterization of the extracellular galactomannan of Aspergillus fumigatus.331. et al. Avian aspergillosis. et al. 3th ed. AUTHORS ADDRESS G Le Loch’ Service de Parasitologie-Mycologie. Bifidobacterium lipoteichoic acid and false ELISA reactivity in Aspergillus antigen detection. REDIG PT. OGLESBEE BL. 10. 363.5433. 62. Maisons-Alfort.65. Disc diffusion testing is a technique with which to assess the antibacterial efficacy of varying agents on isolated organisms. TDDS3. The most common isolates for both groups of birds were coliforms (both lactose and non-lactose fermenting).5%) were Gram negative. These agents were enrofloxacin. Pseudomonas species were recovered from five birds.Nebulisation . Forbes1 BVetMed CBiol MIBiol DipECAMS FRCVS KEYWORDS Disc diffusion test .25mg gentamicin and F10 at dilutions of 1:250 (0. Culture and sensitivity tests were performed. P. 267 . Thirty-five isolates were cultured from the psittaciformes. J.5mg enrofloxacin.Bacterial flora ABSTRACT Nebulisation therapy is widely used in avian medicine for the treatment of respiratory tract infections. using discs containing 0. Chitty2 BVetMed CertZooMed MRCVS. Exeter. This study aimed to establish a method of in-house disc diffusion testing for specific concentrations of nebulising agents and to evaluate the antibacterial efficacy. Thirtyseven bacterial isolates were cultured from the raptors.8%). Fisk3 BSc (Hons). gentamicin and F10. Monks1 BVSc (Hons) MACVSc (Avian Health) CertZooMed MRCVS. Swindon. the efficacy of those dilutions has not been tested against common avian tracheal isolates. at recommended dilutions. C.6%). 1:62. The F10 results were highly variable and the methodology did not appear transferable to this agent. of which 22 (59. tetrasodium ethylene diamine tetraacetic acid (EDTA) and non-toxic ampholytic surfactants. Strathmore Veterinary Clinic2. of which 14 (40%) were Gram positive and 21 (60%) were Gram negative. of three commonly used antibacterial nebulising agents. incorporating healthy and unhealthy raptors and psittacine birds. Although there are published dose rates of commonly nebulised agents. United Kingdom ASSESSMENT OF DISC DIFFUSION TESTS TO EVALUATE THE EFFECTS OF DIFFERENT NEBULISATION AGENTS AGAINST BACTERIA ISOLATED FROM THE AVIAN TRACHEA D. Andover. Zsivanovits1 DMedVet MRCVS. a disinfectant containing quaternary ammonium and biguanide compounds.4%). 1:125 (0.Trachea .Great Western Referrals1. Tracheal swabs were taken from fifty-three birds. Only Enterococci and a single Staphylococcus showed resistance to either enrofloxacin or gentamicin with the methodology used in this study. N.5 (1. Enterococcus and Staphylococcus species.5%) were Gram positive while 15 (40. 0. EDTA disrupts cell wall integrity by chelating metal ions (FOSTER and DEBOER 1998). Aerosol therapy with F10 has been successfully used in cases of lower respiratory tract infections at a dilution of 1:250 and is well tolerated by patients (CHITTY 2002). broad spectrum fluoroquinolone antibiotic that inhibits bacterial DNA-gyrase. bactericidal. tetrasodium ethylene diamine tetraacetic acid (EDTA) and non-toxic ampholytic surfactants. It offers a method of topical administration of antimicrobial agents. acts by interference with bacterial protein synthesis and is bactericidal. Food dye was added to facilitate identification of each disc type. 2 MATERIALS AND METHODS Blank discs were purchased from Tekia2 and sterilised in an autoclave. Gentamicin. Potentially nephrotoxic agents such as gentamicin can be used safely due to limited drug absorption across respiratory membranes. A preliminary test was conducted in which the discs were prepared as above. such as air sacs. gentamicin and F10 were prepared using sterile water as diluent.05ml of solution per disc) and the discs were left to dry before being sent to the laboratoryc. increases hydration of respiratory epithelium and may increase dissolution of necrotic debris. The active constituents of F101 Super Concentrate Disinfectant (F10) include quaternary ammonium and biguanide compounds. the efficacy of those dilutions has not been tested against common tracheal isolates. an aminoglycoside antibiotic. Various concentrations of enrofloxacin. Additionally. to trial disc diffusion testing as a practical means of assessing the efficacy of commonly used nebulising agents (enrofloxacin. quaternary ammonium compounds cause increased permeability of the outer member in Gram negative bacteria (MAILLARD 2002). F10 is said to be virucidal. Activity is concentration dependent and the spectrum of action includes a wide range of Gram negative bacteria. amongst other actions. Aminoglycosides are generally active against Gram negative aerobes including Enterobacteriaceae and Pseudomonas (KROKER 1997). with the exact formulary remaining a commercial secret. The solutions were then placed onto the discs in a sterile fashion (0. Although there are published dose rates of commonly nebulised antimicrobial agents.1 INTRODUCTION Nebulisation therapy is widely used in avian medicine for the treatment of respiratory tract infections. some Gram positive aerobes and organisms such as Mycoplasma and Chlamydophila (KROKER 1997). causing. Nebulisation minimises patient stress by reducing patient handling. The aim of this study was to demonstrate the tracheal bacterial flora in healthy and unhealthy raptors and psittacine birds. Enrofloxacin is a bactericidal. coagulation of cytoplasmic constituents. but lacking food dye and using commercial concentrations of both enrofloxacin (5ug) and gentamicin 268 . Biguanide compounds and quaternary ammonium compounds act via interference with cytoplasmic membrane function. gentamicin and F10) in an in-house setting and to evaluate the efficacy of these agents at recommended concentrations. which may achieve better concentrations than parenteral antimicrobials in organs with poor perfusion. fungicidal and sporicidal (VERWOERD 2001). a filter paper method of F10 testing was used.8%. one Staphylococcus/ Streptococcus selective agar plate and a section of Sabouraud‘s agar. 3 RESULTS Fifty-three birds were sampled. Blood agar was used to estimate total bacterial population. Bacterial growth around and immediately under and over the paper strip was assessed after incubation. gentamicin (5mg/ml) and F10 (1:250) as above and sent to the TDDS laboratoryc. 1. whereas later in the study. and then examined for growth. 19 (39. This gave disc concentrations of 0. Initially. the zones of inhibition diameters were reported in millimetres. Of the 29 psittaciformes. These were taken after mask induction with oxygen isoflurane.6%) originated from raptors and 29 (60. four (21. using commercial culturettes and then sent in transport media to the TDDS laboratoryc. A sterile swab was used to inoculate dried isosensitest plates3 with the ringer solution.b. Disc diffusion testing was performed simultaneously with the experimental and commercial discs and the zones of inhibition were comparable. All solid and liquid media used in the laboratory was supplied by Oxoid Limited3. at 370C for a maximum of 48 hours. Of the remaining 48 samples. Sterile water was used on the filter strips as control. one CLED (cystine lactose electrolyte deficient) agar plate.(10ug). Some samples were subsequently tested with concentrations of F10 (1: 125. The four isolates from healthy raptors were 269 .5. Of the 19 raptors. gentamicin and F101. oxidase and coagulase were carried out to confirm genus and species.4%) from psittacine birds. organisms were only designated as sensitive or resistant. Additionally. This involved placing a strip of filter paper over the inoculated agar plate. and impregnated discs or filter paper were applied to the surface using sterile forceps. ten (34. New discs with added food dye were also tested.6%).5%) were reported as healthy. Further basic identification tests such as Gram stain.25mg and 0. Discs were then prepared using solutions of enrofloxacin (10mg/mL). Tracheal samples were collected on an opportunistic basis at two veterinary centresa. The plates were then examined and colony types were recorded. One of the horse blood agar plates was then incubated under anaerobic conditions and the other four plates were incubated in air.4% for enrofloxacin. The coliform group included lactose-fermenting coliform species (not identified further) and Klebsiella species. Swabs were initially plated onto two 5% columbia whole defibrinated columbia horse blood agar plates. Table 1 shows the distribution of organisms recovered from both raptors and psittaciformes. 0. Plates were incubated overnight at 370C. 1:62. respectively.1%) were reported as healthy. 0. of which five samples did not culture. and wetting it with F10 at identical concentrations to those used in the disc testing. Sensitivity testing was performed by selecting several colonies of a particular species and emulsifying them in 5ml quarter strength ringer solution. with no appreciable difference in effect. The most common isolates are shown in Table 2. Zones of inhibition around the discs were measured and organisms designated sensitive or resistant according to laboratory protocol.5mg. CLED agar was used to identify lactosefermenting isolates and Sabouraud’s agar was used for the isolation of yeast. though. 21/35) of the total isolates. No distinct trends were noted for any variable. two from raptors and six from psittacine birds.3%) and two raptors (20%) showed signs of respiratory disease. Of those birds. Again.6%). three moderate growth) and a Streptococcus species (one very scanty growth).Enterococcus species (one moderate and two heavy growth).2%.8%) Enterococcus isolates showing resistance to at least one antibiotic. The expected tracheal flora of healthy raptors and psittaciformes is not well-documented. 5/29: raptors 21. The majority of birds in our study were unwell.8%) and two birds (28. Pseudomonas and Staphylococcus species as common tracheal isolates (MARTEL et al. One psittacine Staphylococcus isolate was resistant to both enrofloxacin and gentamicin. Nine birds in total had signs of respiratory disease (psittaciforme 17. coliforms (one scanty growth. Enterococcus. Of those birds. There was significant variability in the F10 results. 4 DISCUSSION Although the total distribution of Gram positive and Gram negative bacteria was equal (36/72. these bacteria were also isolated from birds that did not show evidence of respiratory disease. our study found coliforms. there were differences in distribution of raptors and psittaciformes. the zones of inhibition for F10 were variable and unreliable for all seven isolates. showed signs of respiratory disease. Importantly. Some isolates were resistant only to enrofloxacin or gentamicin while some were resistant to both. respectively. Table 3 shows the distribution of resistance against enrofloxacin and gentamicin amongst the isolates obtained. three psittaciformes (50%) and three raptors (25%) showed signs of respiratory disease. Staphylococcus species (moderate growth) and a coliform (heavy growth). between different isolates. There are no guidelines for the determination of appropriate zones of resistance and susceptibility for F10 using disc diffusion test methods. As expected.1%. In agreement with previous studies on unhealthy birds. Klebsiella species (one heavy growth). Enterococcus species accounted for the majority of the resistant isolates. of which three (18. 2004). In fact. which may have biased the results. all seven isolates were resistant to enrofloxacin and gentamicin. Enterococci commonly 270 . The six isolates from healthy psittaciformes were Candida (three scanty growths). coliforms were isolated from four healthy psittaciformes and two healthy raptors while Enterococcus species were isolated from three healthy raptors. different disc concentrations and discs versus filter strips. Psittaciforme birds had Gram negative bacteria comprising over half (60%. with 14 of the 18 (77. Coliforms were isolated from in 16 psittaciformes and seven raptors. Staphylococcus species were isolated from six psittaciformes and ten raptors. whereas 40. Enterococcus species were isolated from six psittaciformes and twelve raptors. A total of eight yeast were isolated. two psittaciformes (33.5% (15/37) of the bacteria isolated from raptors were Gram negative. Enterococcus species are generally considered to be of low pathogenicity and can form part of the normal flora of some mucosal surfaces. 4/19). 50% each). given the much larger concentration of drugs in the in-house discs (0. the presence of Gram positive cocci on tracheal cytology may be a useful marker of potential resistance. including Pseudomonas.za/product_details. ready-to-use’ F10 impregnated discs gave inconsistent results when used in disc diffusion tests. Furthermore. at known bacterial loads according to standard procedures. Only 3/18 (16. then inoculation of agar plates with the broth after a set incubation time. personal communication). It is important to note that the presence of these bacteria was not pathognomonic for clinical signs of respiratory disease in our study. This contrasts to principles of zone inhibition testing.co. The presence of resistance is interesting. 271 . 1999.5mg enrofloxacin and 0. and sensitivity testing is considered clinically important (SELBITZ 1992). without known bacteria loads. That is. 10ug gentamicin). However.25mg gentamicin) compared to the commercial discs (5ug enrofloxacin. To the authors‘ knowledge. the zones of inhibition were measured from the edge of the disc to the edge of the zone in the current study. the aim of this study was to assess the disc diffusion method as commonly used in diverse hospitals and laboratories as an in-house test. Dry. The biocidal label claims of F10 against bacteria have been approved and validated by testing agencies in a number of countries (http://www. Interestingly. Given this pattern. unpublished data using agar diffusion techniques (with the bacteria incorporated into the agar itself) as well as “wet discs” (using the disc diffusion method) has given less variable results (J. showed resistance to enrofloxacin or gentamicin. using whatever bacteria isolated from the trachea. healthandhygiene. the actual concentrations of the enrofloxacin and gentamicin discs were 100 and 25 times more concentrated than commercial discs. Elize Lloyd.asp?ContentId=68&FormDocumentsLi st_Page=3#DocumentsList) Testing usually involves determination the minimal inhibition concentration (MIC) of an agent. the organism is suspended in aqueous solution with the biocide. Another Gram positive coccus. Different disc methodologies and the correlation of MIC with zones of inhibition for F10 are areas worthy of further research. Temperley. YONG et al. However. respectively. while generally zones of inhibition are measured across the diameter of the inhibition zone and the disc. Staphylococcus was the other resistant isolate.exhibit antibiotic resistance. By that. there is no published information about impregnation of quaternary ammonium or biguanide compounds into discs for antibiotic sensitivity testing. 2002). Enrofloxacin and gentamicin discs were produced at concentrations used for nebulisation. The differences in variability may relate to the use of different disc methodology and the use of agar diffusion versus disc diffusion techniques. Studies investigating the use of EDTA-impregnated discs reported conflicting results with poor “appearance and reproducibility” in several strains of bacteria (ARAKAWA et al.7%) Enterococci were sensitive to both enrofloxacin and gentamicin. Enterococcus species accounted for all but one of the resistant isolates in our trial. An appropriate zone of inhibition for a particular biocide requires correlation of the zone of inhibition with the MIC of that biocide. None of the Gram negative isolates in our study. which generally use the lowest concentration that is still effective. The F10 disc diffusion and filter paper methods of testing generated highly variable results. addition of the known concentration into a broth of known antimicrobial concentration. at the concentration that the tested agents are used for nebulisation. 8. FOSTER AP and DEBOER DJ. VERWOERD D. 3.918. VERMINNEN K. California. Bacterial target sites for biocide action. Falco 2001. Pharmak zur Behandlung und Verhutung bakterieller Infektionen In: LOSCHER W. 240. Germany 1992. 5. Monterey. J Clinical Microbiol 2000. MARTEL A. J Clinical Microbiol 2000.18. Proc Assoc Avian Vet. UNGEMACH FR and KROKER R (eds): Pharmakotherapie bei Haus-und Nutztieren. United Kingdom Email: d. 6 CITATION INDEX 1.27. et al. Germany. et al.5 ACKNOWLEDGEMENTS We would like to acknowledge the co-operation we have received from Health and Hygiene (Pty) Ltd and the Microbiology Department of the South African Bureau of Standards. SHIBATA N. ARAKAWA Y. The role of Pseudomonas in canine ear disease. 10: 3798 . et al. LEE K.co. CHITTY J. 92: 16S .monks@gwreferrals. KROKER R. Imipenem-EDTA disk method for differentiation of metallo-B-producing clinical isolates of Pseudomonas spp.uk 272 . 4. YONG D. Swindon. 20(8): 909 . 6. SN1 2NR. SHIBAYAMA K. MAILLARD JY. 25 . 17: 17 . YUM JH. Lehrbuch der veterinarmedizinischen Bakteriologie. 171. AUTHORS ADDRESS Deborah Monks BVSc (Hons) MACVSc (Avian Health) CertZooMed MRCVS Great Western Referrals. and Acinetobacter spp. 2. USA 2002. Proc Assoc Avian Vet. Gustav Fischer Stuttgard. 1997: 211 246.371. Parey Buchverlag Berlin.43. Aerosol use of a novel disinfectant as part of an integrated approach to preventing and treating aspergillosis in falcons in the UAE. J Applied Microbiol Symp Suppl 2002. Microbial pathogens associated with respiratory disorders in psittacine birds.27S SELBITZ HJ. A novel disinfectant in psittacine respiratory disease. Convenient test for screening metallo-B-lactamase-producing Gram-negative bacteria by using thiol compounds. 9. RAUE R. 7. New Orleans 2004: 369 . County Park Estate.3801. Shrivenham Road. Comp Cont Ed 1998. 38(1): 40 . Yeast Raptors Psittaciformes Total 2 6 8 Gram positive (% of total) 22 (59.5) 14 (40) 36 (50) Bacteria Gram negative (% of total) 15 (40.7 33.9 (Gram �) Staphylococcus 6 42. Number % gram isolates + isolates Raptors: Coliforms 7 NA N=22 gram positive Pseudomonas 5 NA (Gram +) N=15 gram negative Enterococcus 12 54. Common isolates by taxonomic group.3 NA NA 76.7 2.Table 1.9 17. Sensitivity spectrum for enrofloxacin and gentamicin.9 % of gram � isolates 46.5) 21 (60) 36 (50) Total isolates 37 35 72 Table 2.8 NA NA % of total isolates 18.5 Psittaciformes: Coliforms 16 NA N=14 gram positive Pseudomonas 1 NA (Gram +) N=21 gram negative Enterococcus 6 42.2 4.0 45.5 32. Summary of isolated organisms.9 13. Enrofloxacin Gentamicin Sensitive Resistant Sensitive Resistant Raptors: Coliforms 7 0 7 0 Pseudomonas 5 0 5 0 Enterococcus 6 6 5 6 Staphylococcus 10 0 9 0 Total Gram 16 6 16 6 Positive Total Gram Negative 15 0 15 0 Psittaciformes: Coliforms 16 0 16 0 Pseudomonas 1 0 1 0 Enterococcus 5 1 3 3 Staphylococcus 5 1* 5 1* Total Gram 12 2 10 4 Positive Total Gram 21 0 21 0 Negative Total 7 5 11 9 22 5 16 1 6 6 14 21 273 .1 Table 3.1 17.5 (Gram �) Staphylococcus 10 45.4 27. is common in rotavirus infection. DVM.Kimron Veterinary Institute. Lublin. V. The virus was diagnosed by ELISA only in 3 of 18 suspected birds (two grey parrots and one ring-necked parakeet). could not be demonstrated. Rotavirus have been found especially in intestinal mucosal cells. 1 INTRODUCTION Rotavirus is a double-stranded RNA virus of the Reoviridae family with a three-layer capsid 70-80 nm in diameter lacking an external envelope. also by immuno-electron microscopy-negative staining. Rotavirus invades the intestinal mucosal cells. Eight of 18 flocks (mostly broilers) with signs of stunting. Clinical signs of rotavirus infection include growth retardation.e. but there is no consensus about cross infection between the two groups. especially at the edges of the intestinal villi. Immunosuppression and outbreaks of other intestinal pathogens such as Clostridium sp. DVM. lack of uniformity in flock.. lack of uniformity in a flock. S. but also in two birds without clinical suspicion (grey parrot. Israel DIAGNOSES OF ROTAVIRUS IN PET BIRDS VS POULTRY A. The main replication site is in the enterocytes of the small intestines. In broilers we found association between rotavirus infection and malabsorption syndrome. In pet birds. i. as had been suggested by some researchers. The virus is one of the causes of enteritis with diarrhoea in birds. lamina propria. It seems that rotavirus is more abundant in stunted poultry than in pet birds with similar signs. abnormal feathering and sometimes bone lesions. PhD. Bet Dagan. but also in the colon and cecum. Viral replication causes lysis of the host cells and impairing of absorption. Mechani. cockatoo). as in mammals. diarrhoea. while association with reovirus infection. DVM KEYWORDS Rotavirus – Malabsorption syndrome – VP6 . and in few of the cases. 274 . low food conversion. rotavirus is one of the causes to gut dilatation and impaired food absorption. but also in the inner connective tissue of the mucosa. undigested food and dilatation of the proventriculus were positive to the major inner capsid protein VP6 of group A-rotavirus by ELISA and by immunohistochemistry. Bumbarov. and inside intestinal contents. Infection by rotavirus includes growth retardation. anaemia.Immunohistochemistry ABSTRACT Rotavirus is one of the causes of enteritis in birds. and by immuno-electron microscopy with rotavirus antiserum. Thus. on average 37% of the manure. The main pathological lesions of the syndrome consist of white-coloured or transparent intestinal walls. 1. enlarged gall bladder. abnormal feathering and sometimes bone lesions. but the results were not satisfactory. or coccidia. The most important portal of entry is by ingestion. in 42% of the cases. In a few of the cases the virus was also diagnosed by electron microscopy using a negative staining technique. in comparison to 15% of cases in duodenum and 29% in jejunum-ileum. breeders.05). Most reports are derived from poultry and less information is known about pet birds. diarrhoea.e. the highest rate of infection was found in cecum. Rotavirus was also detected by ELISA in poultry manure. pancreatic atrophy. Immunosuppression and outbreaks of other intestinal pathogens such as Clostridium sp. Virus prevalence was relatively high in turkey manure (about 40%). In contrast. Clinical disease (retarded growth with or without diarrhoea) was reproduced in 12 inoculation experiments with filtered intestinal contents after direct administration into the crop of specific pathogen free 1-day-old chicks. Diagnosis of the virus by immunofluorescence was also tested. Serological surveys of reovirus in flocks were negative in an experimental model in which infected intestinal contents from retarded birds were inoculated into specific pathogen free 1–day-old chicks and follow-up for antibody titres in affected birds (as most of the inoculated chicks became). could be detected by ELISA and by immunohistochemistry. is its main aetiology.04. One of the advantages in using immunohistochemistry is the ability to locate the virus in tissue (as red colour appears at the sites of antigen localization). making the excreted faeces a route of spreading the virus. In testing the various segments of the intestines for rotavirus infection. The virus was also observed in intestinal contents. sometimes proventriculitis. The high prevalence in manure is probably due to its accumulation in manure and its high resistance in the environment 275 .1.low food conversion. anaemia. Rotavirus was diagnosed particularly in intestinal mucosal cells and to a lesser extent in the inner connective tissue of the mucosa i. sensitivity to rotavirus infection is limited to broiler chicks younger than 1-2 days. Rotavirus in poultry In broilers we found an association between rotavirus infection and malabsorption syndrome. lamina propria. reovirus either rotavirus. P<0. in 8 out of 18 flocks (mostly broilers) with signs that may indicate rotavirus. The syndrome could not be reproduced in chicks older than 2 days. degeneration of the bursa of Fabricius and rickets. layers. a group A-rotavirus antigen – major inner capsid protein VP6. with significant correlation between the two diagnostic methods (chi-square coefficient 4. that according various researchers. The virus is excreted in high numbers in faeces and can be transmitted directly and indirectly. are common in rotavirus infection. Sixteen tested flocks out of 21 received the same result with both methods. but was found also in manure from broilers. pullets and “organic chickens”. Inoculation of turkey poults did not reproduce the disease. Rotavirus was also detected in two birds without clinical suspicion (grey parrot. POB 12. therefore more research needs to be done in pet birds. proventricular dilatation and undigested food as in other cases. AUTHORS ADDRESS A. 2 CONCLUSIONS 1. In pet birds rotavirus is less familiar than in domesticated birds. PhD Division of Avian & Fish Diseases. Watery diarrhoea may occur in infected pigeons. 2. widespread especially in lovebirds and pigeons. rotavirus was diagnosed by ELISA only in 3 of 18 birds suspected according to clinical and pathological signs: two grey parrots (Psittacus erithacus) with pathological presentation of proventricular dilatation disease (PDD) and a ringnecked parakeet (Psittacula krameri) from a flock with respiratory disturbances and mortality. In poultry rotavirus is usually associated with a clinical manifestation of diarrhoea/stunting and in pet birds this association is less obvious.gov. Israel Email: lublina@int. 3. cockatoo). Some of the strains do not cause clinical symptoms in carrier birds.il 276 .1.Bet Dagan 50250. Rotavirus is more abundant in stunted poultry than in pet birds with similar signs. In a survey conducted in our laboratory. More information concerning rotavirus in poultry exists compared with pet birds. DVM. Lublin. Kimron Veterinary Institute. while necropsy findings in this case included diarrhoea.2. Rotavirus in pet birds In pet birds rotavirus is one of the causes of gut dilatation and impaired food absorption. the designation psittacine adenovirus (PsAdV) is proposed. Therefore.L1 region ABSTRACT Adenovirus infections in psittacine birds have been well known and were mainly caused by fowl adenoviruses (FAdV).. 27 aa are located in the 4 hypervariable regions of loop 1. egg drop syndrome virus and hemorrhagic enteritis virus.3 and 61. Out of these. A PCR product of approximately 590 bp in size was amplified and sequenced. H. the percentage of identical nucleotides ranged between 60. Germany PHYLOGENETIC ANALYSIS OF THE HEXON LOOP 1 REGION OF ADENOVIRUS ISOLATED FROM PSITTACINE BIRDS INDICATES THE EXISTENCE OF A NEW PSITTACINE ADENOVIRUS R. Faculty of Veterinary Medicine. Gerlach2 and H.Hexon gene . Leipzig.PsAdV .3 and 67. 1 INTRODUCTION The classification of the family Adenoviridae is currently under consideration.0. A PCR amplifying the variable loop 1 region of the hexon gene was developed using primers located in two conserved pedestal regions. Raue1. Munich2. Avian adenoviruses characterised by a common group-specific antigen were previously classified as group I and still will represent the genus Aviadenovirus. indicating that the sequence originated from a new avian adenovirus. University of Leipzig. 37 unique aa exchanges were observed. Müller1 KEYWORDS Psittacine adenovirus . showing morphological and histological signs of an adenovirus infection. In this region. In a phylogenetical analysis the sequence obtained grouped outside of the FAdV reference strains of all 12 serotypes. which encode the serotype-specific epitopes. Further group I 277 .0. According to the classification of the International Committee on Taxonomy of Viruses.PCR . and that of identical amino acids (aa) between 51. were investigated in this study. Prototypes of this genus are the fowl adenoviruses (FAdV) with 12 serotypes. Liver samples collected from an outbreak of acute disease in Poicephalus spp.Institute for Virology1. In a comparison to the FAdV reference strains. it is obvious that a new conventional adenovirus from psittacine birds has been identified. The capsid consists of 252 capsomers. the fibre and the hexon. two conserved regions were identified for primer annealing and amplification of the L1 region of the hexon gene. duck and pigeon. With a length of more than 130 amino acids (aa) it is the largest one. particularly of L1 and L2. The sense primer Hex L1-s (5´-atgggagcsacctayttcgacat-3´) is located in a region from nt 301 to nt 323 of the hexon gene of FAdV1.avian adenoviruses have been isolated from turkey. By the alignment of published nt sequences of all 12 FAdV serotypes. with acute disease. 4 HVR in L1. L2 and L4 are located. 1986). On the outer surface of the hexon 3 loop (L) regions designated as L1. The L3 region combines the outer surface regions and the conserved pedestal (P) regions P1 and P2 forming the inner surface. the antisense primer Hex L1-as (5´-aaattgtccckraanccgatgta-3´) corresponds to a region spanning nt 890 to nt 868. the phylogenetic analysis of the sequenced L1 region showed remarkable differences supporting the proposal for a new psittacine adenovirus (PsAdV). total DNA was isolated using the DNeasy Tissue Kit (Qiagen. Seven hyper variable regions (HVR) were identified in the L regions of the hexon gene. Adenoviruses are non-enveloped particles of 70-90 nm in diameter with a single linear molecule of double-stranded DNA of variable size between approximately 26 and 45 kbp (BENKÖ et al. Liver samples (M158-04 and M159-04) were collected and investigated by histological examination. all FAdV strains have two fibres at each penton base. 2 MATERIALS AND METHODS Two 42 days-old Senegal parrots (Poicephalus senegalus) hatched out in an aviary died within 24 hours after the onset of severe clinical disease demonstrated by ruffled feathers and anorexia. 2 in L2 and 1 in L4 (CRAWFORD-MIKSZA and SCHNURR 1996). 2000). In a comparison with various avian adenovirus reference strains. Intranuclear eosinophilic inclusion bodies in hepatocytes of these samples indicate the presence of an adenovirus. In this study liver samples showing typical histological signs of an adenovirus infection were collected from Poicephalus spp. Hilden. 278 .5% homology between FAdV1 and FAdV10. From the samples. Germany) as recommended by the supplier. A PCR amplifying the L1 region of the hexon gene was developed. The expected fragment size was approximately 590 bp. Serotype-specific determinants are located on both. respectively. It has been demonstrated that the type-specific antigenic determinant � is encoded by the HVR. The former group II egg drop syndrome virus (EDSV) and group III turkey adenovirus 3 (TAdV-3) recently have been moved to the new genera Siadenovirus and Atadenovirus. The nucleotide (nt) sequence obtained was analysed at the nt and the aa level. The three-dimensional structure of human adenovirus 2 has been identified (ROBERTS et al. The fibre is bound non-covalently to the penton base. As demonstrated by the analysis of L regions of different avian adenoviruses. which can be differentiated by cross neutralization tests. As a particularity. 240 hexons and 12 pentons. L1 represents the most variable region (RAUE and HESS 1998). showing only 42. Erlangen.Isolated DNA was tested for the presence of adenovirus DNA by PCR as follows: 5 units of Taq-DNA-polymerase (Peqlab. cloned in to the pCR4-TOPO® vector as recommended by the supplier (Invitrogen. The nt sequence was determined using both M13 and M13 reverse primers and an automated 377 ABI DNA Sequencer together with the Dye Terminator Cycle Sequencing Kit (Applied Biosystems Inc. The PCR was started with an initial denaturation step of 5 min at 95 °C. Foster City.5 %. USA). 30 s at 56 °C for primer annealing and 45 s at 72 °C for elongation. Erlangen. the sequence of EDSV showed the lowest percentage of identity when compared with that of M158-04. whereas that of the TAdV-3 sequence and M158-04 was only 14. The temperature profile of the following 40 cycles consisted of 30 s at 94 °C for denaturation. M159-04) whereas sample 107-04 reacted negative. The overall percentage of identity among the sequences obtained from the FAdV reference strains and M158-04 ranged between 60. 10 mM of each dNTP (Peqlab. The PCR product of liver sample M158-04 was purified (Qiaquick gel purification kit...55). The reaction was terminated by a final elongation step of 5 min at 72 °C. This bird did not show any clinical or pathological signs of an adenovirus infection. 279 . a liver sample (107-04) taken from a 78 days-old Jardin’s parrot (Poicephalus gulielmi) was used.5% agarose gel stained with ethidium bromide and visualised by UV transillumination. Using the primer combination Hex L1-s and Hex L1-as. Both samples reacting positive in the PCR showed the same restriction enzyme pattern after cleavage with different restriction enzymes. were aligned with the published sequences of the reference strains of avian adenoviruses using the Clustal W method of the MegAlign program (DNA Star Inc.1 %. 160 mM (NH4)2S04 and 20 mM MgCl). 3 RESULTS For the specificity testing of the developed PCR. Germany). Germany) and sequenced for phylogenetic analysis. the presence of adenoviral DNA was demonstrated in fresh liver samples collected from two Senegal parrots (Poicephalus senegalus) with clinical signs of an adenovirus infection (M158-04. Germany) and 100 pmol of primers Hex L1-s and Hex L1-as were mixed with 4 µl of template DNA in a total volume of 50 µl. Therefore. The Nt sequence of the 587 bp PCR products and the deduced aa sequence. Karlsruhe. The length of the fragment compared varied between 575 nt (strain TAdV-3) and 602 nt (strains 340 and IBH-2A). With 12. 5 µl of 10x reaction buffer (200 mM Tris-HCl (pH 8. USA). Hilden. only the nt sequence of the PCR product obtained with sample M158-04 was established and compared with the sequences of other avian adenoviruses. The PCR products (5 µl aliquots) were separated on a 1. Germany).3 to 67. Qiagen.0 %. Wisconsin. but had died following acute psittacine beak and feather disease. 4 DISCUSSION This is the first report of a specific psittacine adenovirus detected by PCR from Senegal parrots with signs of acute disease.3 to 61. The alignment of deduced aa sequences revealed that unique aa exchanges were observed at 30 positions in the M158-04 sequence: twelve of these were located in the first variable region A1. the existence of FAdV2. a fast and convenient method for the characterisation of avian adenoviruses became available. Furthermore. Since it was demonstrated recently that FAdV strains can be typed by their hexon L1 sequences (MEULEMANS et al. As this region encodes most of the serotype-specific epitopes. FAdV4 and FAdV8. 5 in A2. 1). respectively.5 %.81-59. it may be expected from the deduced amino acid sequence to show whether a new serotype exists or not. 7 aa of the M158-04 sequence were identical to those in the EDSV and TAdV-3 sequences.99 %. the M158-04 sequence was a g/c-rich sequence and within the range observed in the FAdV sequences (48. whereas that of the sequences obtained from EDSV and M158-04 was only 34. With a g/c content of 52. 1) showing 3 major branches for the proposed genera. 2001). Both. Atadenovirus (EDSV) and Siadenovirus (TAdV-3). Six out of the seven branches formed the groups representing the genogroups as published (ZSAK and KISARY 1984). FAdV3.A phylogenetic tree was established (Fig. within this major branch a sole minor branch was formed by M158-04 indicating that M158-04 represents a new avian adenovirus genotype. Avian adenoviruses have been detected in psittacine birds by electron microscopy many times. The length of the aa sequences varied between 191 (strain TAdV-3) and 200 aa (strains 340 and IBH-2A). However. only limited data exist on the classification of adenoviruses infecting psittacines. the sequence of TAdV-3 showed the lowest percentage of identity as compared to sequence of M158-04. was demonstrated in psittacines with or without clinical signs by neutralisation tests. but not to those observed in the sequences of the FAdV strains. The phylogenetic tree of the deduced aa sequences was comparable to the one described for the nt sequences. It is evident that the sequence M158-04 represents a new group I avian adenovirus which should be designated as PsAdV. Within the Aviadenovirus branch. a PCR protocol was developed amplifying the L1 region of the hexon gene. 7 minor branches were observed. 1 in A2 and 1 in A3) and 2 in the conserved regions.0 %. The charge at pH 7. fife out of these were located in the variable regions A1-A4 (3 in A1. The evidence that the M158-04 sequence belongs to the group I avian adenoviruses is supported by the analysis of its g/c content. the sequences of EDSV and TAdV-3 showed lower g/c contents. The sequence obtained here revealed significant differences on the nt as well as on the deduced aa level (Fig. In accordance with the 280 . Therefore. With 32. So far. 4 in A3. The overall percentage of identity among the sequences obtained from the FAdV reference strains and M158-04 ranged between 51.0 and the isoelectric point of the M158-04 sequence are in a range observed in the case of the FAdV strains. respectively.03 %). However. Aviadenovirus (FAdV1-12).0 %. 1 in A4 and 8 in the conserved regions. Intervirol 1984. The identification of new adenoviruses by sequencing is a common technique.114 2. 7. Furthermore.nomenclature used by the International Committee of Taxonomy of Viruses (ICTV) it is proposed to designate this new avian adenovirus as PsAdV.1151 ZSAK L. Science 1986: 232: 1148 . J Virol 1996. 227 . nothing is known about the distribution and significance of PsAdV and its possible role for a specific disease in psittacines. Three-dimensional structure of the adenovirus major coat protein hexon.238 CRAWFORD-MIKSZA LK. isoelectric point and charge at pH 7. On the other side. San Diego: Academic Press 2000.e. 281 . M. (eds): Virus Taxonomy. PsAdV shows common characteristics of group I avian adenovirus such as its g/c content. sera of birds living in the same aviary as those investigated here and surviving the disease reacted positive with common group I avian adenovirus-specific antigens in an immunodiffusion test (H. 73: 211 . So far. 30: 655 . 5. Polymerase chain reaction combined with restriction enzyme analysis for detection and differentiation of fowl adenoviruses.660 RAUE R.34 MEULEMANS G. Hexon based PCRs combined with restriction enzyme analysis for rapid detection and differentiation of fowl adenoviruses and egg drop syndrome virus. Seventh Report of the International Committee on Taxonomy of Viruses. Avian Pathol 2001. 4.1844 KOVACS GM.217 ROBERTS MM. 37 out of 195 (19. The PCR described here is a suitable and convenient method to identify PsAdV DNA in liver samples. In: VAN REGENMORTEL MHV. 27 out of them were located in the 4 hypervariable regions of loop 1 carrying most of the serotype-specific epitopes. 2003). J Virol Methods 1998. HARRACH B and RUSSELL WC. i. Here. 6. Grouping of fowl adenoviruses based upon the restriction patterns of DNA generated by BamHI and HindIII. the sequence obtained allowed the development of specific and sensitive PCR protocols which can be used for epidemiological studies in the future. BISHOP DHL. 22: 110 . Hafez. Analysis of 15 adenovirus hexon proteins reveals the location and structure of seven hypervariable regions containing serotype-specific residues. Furthermore. unpublished data).0 %) aa of the PsAdV sequence were not observed in any of the FAdV reference strains. Phylogenetic analysis of the hexon and protease genes of a fish adenovirus isolated from white sturgeon (Acipenser transmontanus) supports the proposal for a new adenovirus genus. an adenovirus from white sturgeon was recently identified by its hexon sequence and based on that a new adenovirus group was predicted (KOVACS et al. 98: 27 . 5 CITATION INDEX 1. Virus Res 2003.0. BENKÖ M. 3. 70: 1836 . FAUQUET CM. et al. These observations might indicate that PsAdV is a new avian adenovirus with special (antigenic) characteristics. Family Adenoviridae. Phylogenetic tree established with the nucleotide sequences of the approximately 590 bp fragment of the hexon gene containing the L1 region (clustal W method). D-04103 Leipzig. 282 . Numbers in parenthesis correspond to the accession numbers. Germany Email:
[email protected] Fig. An den Tierkliniken 29. Typing of FAdV into genogroups (A-E) according to ZSAK and KISARY (1984) is also mentioned. Faculty of Veterinary Medicine. Rüdiger Raue Institute for Virology.AUTHORS ADRESS Dr. The box indicates sequences belonging to group I avian adenovirus classified as Aviadenovirus. 1.uni-leipzig. University Leipzig. these variations lead to difficulties in the interpretation of results and are therefore responsible for the under use of plasma protein electrophoresis in avian medicine. 2000).Parc de Clères Jean Delacour Muséum National d’Histoire Naturelle. 34 black kites (Milvus migrans migrans. MsC. Y. 18 blacksmith lapwings (Vanellus armatus) were blood sampled to establish reference ranges and to compare interspecific differences. 36 rock doves (Columbia livia domestica). KEYWORDS Protein – Electrophoresis – Reference ranges. Ecole Nationale Vétérinaire de Nantes.Duvernoy. 44 jackdaws (Corvus monedula). most of the data available in the literature deals with raptors or psittacine birds and there is a lack of information 283 . France PLASMA ELECTROPHORESIS REFERENCE RANGES IN VARIOUS BIRD SPECIES D. Roman . PhD. 40 peafowls (Pavo cristatus). Unfortunately. 42 white storks (Ciconia ciconia). In the daily practice. at this time. DVM. Chaste . Practitioners should be aware of the species reference ranges to be confident in their results and to interpret variations of the different fractions concentrations. However. Milvus migrans parasitus). D. insisting on peculiarities of each species. M. Ordonneau. wide variations of electrophoretic patterns have been described between some bird taxa (BLANCO and HOFLE 2003. ZAIAS et al. 291 birds belonging to 8 different species (62 bar headed geese (Anser indicus). Unfortunately. ABSTRACT Plasma protein electrophoresis is generally recognised to be a very reliable diagnostic tool in birds for many pathologic conditions. CRAY and TATUM 1998). Biol. 13 African grey parrots (Psittacus erithacus). 1 INTRODUCTION Plasma protein electrophoresis is generally recognised to be a very reliable diagnostic tool in birds for many pathologic conditions (WERNER and REAVILL 1999.C. Vet student. CRAY and TATUM 1998. This study focuses on plasma protein electrophoresis of birds belonging to species that are not well documented. Bomsel. wide variations of electrophoretic patterns have already been described between some bird taxa. because it is less prone to haemolysis and contains fibrinogen. Heparinised blood was immediately centrifuged and plasma samples were stored in cryotubes at -20°C for a variable duration before being send to the laboratory.about other bird taxa. Samples were then allowed to thaw and were rehomogenised 1 hour before analysis. presumed healthy. The electrophoresis curves and dosages of the different fractions were 284 . Total protein concentration was determined by the biuret method with the Roche integra system. 1 to 5 ml of blood was collected from the right jugular or brachial vein. transferrin. Plasma aliquots were loaded onto the gel and allowed to migrate for 8 min at 296 V (6mA) in a Sebia Hydrasis system. 44 free ranging jackdaws (Corvus monedula) from the zoological park of Clères (epidemiological study). 1996). 36 free ranging rock doves (Columbia livia domestica) from the Ménagerie du jardin des plantes de Paris (France) (epidemiological study). Agarose gel electrophoresis was performed with Hygragel protein 15/30 kits (Sebia. these variations lead to difficulties in the interpretation of the results and are therefore responsible for the under use of plasma protein electrophoresis in bird medicine. depending on the species. 40 peafowls (Pavo cristatus) from the zoological park of Clères. WERNER and REAVILL 1999). in order to minimise intraspecific variations. birds belonging to 8 phylogenetically distant species: 62 bar headed geese (Anser indicus) from the zoological parks of Clères (France) and Branféré (France). La Haute Touche (France) and Branféré. immunoglobulins in �-globulins) (CRAY et al.13 African grey parrots (Psittacus erithacus) from the zoological park of Clères. which is a characteristic acute phase protein (KANEKO 1989. In daily practice. Some data is available about the protein content of some fractions in psittacine birds (albumin. Blood was therefore collected in lithium heparin. The purpose of the study reported here was to define plasma protein electrophoresis reference ranges on various bird species in order to extend the use of this test to other avian species and to comment on interspecific differences. but much of it seems to have been extrapolated from mammals (KANEKO 1989). care was taken to avoid haemolysis by taking blood samples with 20 G needles. and some private owners. �-antitrypsin. fibrinogen. From each bird. we studied the largest population sample as possible. the variations between bird taxa make it difficult to define the different protein fractions in a standardised way. haptoglobulin in �-globulins. Once dried and coloured with amidoblack in the same system. gels were scanned by a high resolution Epson expression 1680pro flat scanner. 34 black kites (Milvus migrans migrans and parasitus) from the falcon house of the Puy du Fou (France). Furthermore. 2 MATERIALS AND METHODS This study was conducted on 291 adult. since it is commonly admitted that plasma is preferable to serum for protein electrophoresis in birds. 42 white storks (Ciconia ciconia) from the zoological parks of Clères. the Ménagerie du jardin des plantes. France). complement. Since haemoglobin is known to migrate in the � fraction (WERNER and REAVILL 1999). Protein electrophoresis was carried out on plasma. 18 blacksmith lapwings (Vanellus armatus) from the zoological park of Clères. �-lipoprotein in �-globulins. In each species. �-lipoprotein. �2-macroglobulin. Z = 41.001) and all protein fractions (Kruskal and Wallis test: df = 6. albumin / globulin ratio (Kruskal and Wallis’ test: df = 6. p < 0. Prealbumin fraction values were very low and were included to albumin fraction values. a very important peak in �2-fraction. the blacksmith lapwing showed one peak for each fraction except for �1-fraction. Systat 7.001).141. we used basic statistic and Kruskal and Wallis’ one way analysis of variance. The peafowl’s curve was characterised by 2 peaks in the �2-fraction. Finally. The African grey parrot’s curve had the same characteristics as the rock dove’s with a smaller peak in �2 and �-fractions. This stresses the importance of defining reference ranges in main taxonomic groups. it is generally accepted that the most relevant parameters for the interpretation of avian proteinograms are total protein concentration and albumin / globulin ratio (FUDGE 2000). �2-globulin. �1-globulin. The bar headed goose’s curve was very simple with one peak for each fraction. there was one peak for each fraction and a tray between �2-fraction and �-fraction. �-globulin) (Fig. considering the fact that fibrinogen seems to be the only protein migrating in a consistent way. p < 0. 4 DISCUSSION This study highlights that there is no intraspecific variability for the parameters studied. The rock dove’s curve was characterised by an unapparent �1-fraction.made with Phoresis Sebia’s software. 1-8). Without any standards for defining protein fractions. No intraspecific differences were found whatever the species may be (Kruskal and Wallis’ test). we observed an unapparent �1-fraction and 2 peaks in �-fraction. p ≤ 0. 285 .001). However. Anatidae. In the black kite’s curve. Columbidae) (ORDONNEAU unpublished data). Interspecific differences were highly significant for all parameters: total protein concentration (Kruskal and Wallis’ test: df = 6. 1-8). whatever the species may be (ORDONNEAU unpublished data). Results are documented in Table 1. �-globulin. Electrophoretic patterns appeared to be divided in 5 fractions (. In the jackdaw’s curve.0 software (SPSS INC© 1997) was used for all analyses. Z = 193. in a defined taxon. and 2 peaks in the �-fraction. the fractions were defined by comparison with other bird species. electrophoretic patterns of different species generally share many resemblances. The white stork’s curve was similar.Albumin. showing that there is a great homogeneity within taxonomic groups (Phasianidae.724. which was unapparent. Considering the small size of the population studied. 3 RESULTS Without any information about a standardized method of defining the different protein fractions. In this study. these parameters appeared to be species specific. Electrophoretic patterns and values are completely species specific (Fig. with a higher �1-fraction. and that albumin fraction was the first high peak in the electrophoretic patterns. This result is quite surprising and was not documented in other publications about this species (LUMEIJ and BRUIJNE 1987). by 2D electrophoresis and mass spectroscopy. Proc Assoc Avian Vet. Applications of protein electrophoresis in avian diagnostics. Electrophorèse et Immunofixation des protéines sériques. BOSSART GD and HARRIS D. Further studies will focus on the identification of this important protein fraction in Columbidae. he cannot be confident with the interpretation of results. Without this knowledge. This difference could be related to the use of different kinds of gels (cellulose acetate instead of agarose). CRAY C and TATUM LM. Due to important taxonomic differences. LE CARRER D. However. In: KANEKO JJ. different ways of defining protein fractions could have led to different results.In the rock dove’s curve. 4. Tenerife 2003: 256 . the black kite’s and African grey parrot’s data appeared to be similar to information about raptors (BLANCO and HOFLE 2003) and African grey parrot (CRAY and TATUM 1998). Serum proteins and the dysproteinemias. Laboratoires SEBIA. BLANCO JM and HOFLE U. the use of the Sebia system enabled us to use gels that were the same. 4th ed. Tampa 1996: 97 . but it often appears to have been extrapolated from mammal serum protein electrophoresis (KANEKO 1989).261. without this information. 1998. The practitioner should always pay attention to the technique used in his usual laboratory to be able to compare his own results with published reference ranges (CRAY and TATUM 1998). (ed): Clinical Biochemistry of Domestic Animals. and there was no variation related to the operator’s handling. publication of reference values is not useful for the bird practitioner. Some data was published about the composition of protein fractions. 12(1): 4 . it appears very difficult to define a standardized way of defining protein fractions in birds. Proc Euro Assoc Avian Vet. 5 CITATION INDEX 1.10.165. Plasma protein electrophoresis as diagnostic an prognostic tool in Raptors. KANEKO JJ. 3. 2. J Avian Med Surg 1998. In this study. authors should always illustrate the way they defined each protein fraction. the practitioner must be very familiar with the electrophoretic patterns of bird species he deals with. San Diego: Academic Press Inc. Further studies will focus on identification of proteins that make up each fraction. Plasma protein electrophoresis : an update. 142 . In further studies. Identification of these proteins should allow better definition of protein fractions in order to standardise plasma protein electrophoresis in various bird taxa. Without this information. Agarose gels are more discriminating and allow a better separation of protein fractions (LE CARRER 1998). 5. 1989. �2-fraction concentration was higher than albumin fraction concentration. 286 . At this time. CRAY C. One must choose a standardised technique to allow comparison between further studies. In this study.100. and specially J. N. Avian Pathol 1987. Huyghe from the zoological park of Branféré. and biochemical profiles of brown pelicans (Pelecanus occidentalis). plasma protein. 8. Chai. Vet student N°11 Résidence Plein Sud. WERNER LL and REAVILL DR. Blood chemistry reference values in racing pigeons (Columbia livia domestica). 7. France Email: ordonneau. Am J Vet Res 2000. (ed): Laboratory Medicine : Avian and Exotic Pets.do@voila. Hematologic.774.408. FOX WP. Av d’Aunis.L. 9. 14: 401 . In: FUDGE AM. ZAIAS J. the falcon house of the Puy du Fou. from the zoological park of la Haute Touche and F..173. Ordonneau. 171 . Avian protein disorders.662. ROSENTHAL K. 6 AKNOWLEDGMENTS We thank Sebia France for their generous gift of Hydragel protein 15/30 kits. AUTHORS ADRESS D.fr 287 . CRAY C and ALTMAN NH. from the Ménagerie du jardin des plantes. LUMEIJ JT and BRUIJNE JJ.6. Liégeois. 61(7): 771 . Philadelphia: WB Saunders 2000. 17570 La Palmyre.L. The diagnostic utility of serum protein electrophoresis. 2: 651 . Vet Clin North Amer: Exot Anim Prac 1999. 67 0.99 2.34 25.08 6.78 0.13 7.15 0.21 0.16 Total protein ± 5.46 1.76 0.11 n=18 % A/G* = albumin / globulin (�1+�2+�+�) ratio A/G* = albumin / globulin (�1+�2+�+�) ratio .08 5.21 0.77 1.91 8.86 2 1.59 1.22 43.04 4.54 3.59 3.64 13.65 1.09 0.57 0.18 n=34 % Pavo cristatus g/L 44.39 4.31 0.98 15.32 26.14 17.45 1.8 4.88 2.94 29.7 25.29 2.94 3.07 53.23 0.32 2.24 0.67 1.01 5.66 2.29 2.05 Albumin Alpha 1 Alpha 2 Beta Gamma ± 0.6 3.1 2.64 2.34 3.58 29.01 4.04 4.1 4.39 0.82 5.13 24.7 6.32 5.02 3.68 1.54 0.4 5.76 0.56 0.08 2.9 0.16 15.2 2.14 6.31 1.34 57.07 23.93 2.34 6.3 6.55 2.23 2.27 0.12 0.45 39.11 2.7 8.23 2.73 4.24 ± mean ± mean ± mean ± mean ± mean 2.71 15.08 16.59 1.23 5.Table 1 : Plasma protein electrophoresis mean values and SD in g/L and percentages of healthy birds of 8 different species A/G* mean 1.02 3.59 2.06 1.86 1.11 mean Anser indicus g/L 41.03 5.01 2.2 25.22 2.94 9.28 39.07 0.14 15.51 12.98 21.79 5.84 1.6 0.86 0.38 0.02 4.38 n=40 % Psittacus erithacus g/L 37.99 2.14 3.51 36.91 1.12 4.76 1.87 10.19 22.3 1.66 4.17 n=13 % Vanellus armatus g/L 36.21 n=42 % Columbia livia domestica g/L 36.34 10.67 1.16 n=62 % Ciconia ciconia g/L 40.98 4.75 2.52 1.91 7.15 0.3 13.18 13.64 3.41 9.94 5.48 4.63 1.47 0.96 5.11 10.15 14.33 18.87 1.79 0.44 1.76 8.82 0.5 2.39 2.25 0.83 n=36 % 288 Corvus monedula g/L 36 n=44 % Milvus migrans g/L 40.21 2.55 7.52 16.17 2.1 7.39 17.2 19.87 2.27 1.92 10.42 0.28 0.71 1.81 9.42 0.75 5.68 2.65 2.7 3.09 1.36 6.67 0.48 2.77 9.95 2.02 2.99 2.45 1.14 0.89 1.51 42.57 1.88 0.95 5.46 56. 289 . Changes on the electrophoretic patterns occurred mainly in the � fraction .Electrophoresis .DUVERNOY. MSc. The inflammatory condition led to visible changes as early as 3 hours after surgery for haematology and 16 hours after surgery for protein electrophoresis. INTRODUCTION Serum protein electrophoresis has been extensively used for 3 decades in human diagnostics. ORDONNEAU. BOMSEL. ROMAN. 28 and 52 hours after surgery. Like haematology.C.and in the � fraction. 16. In acute phases of diseases. CAMPBELL 1995). Muséum National d’Histoire Naturelle. PhD. 11. DVM.Inflammation . 6. protein electrophoresis is quite a non specific laboratory examination which is thought to react rapidly. PhD. significant changes in the electrophoretogram may already be recorded (CRAY and 290 . M. D. but its use in avian medicine is far more recent. 1. KEYWORDS Protein .where fibrinogen is known to migrate .Parc de Clères Jean Delacour. MSc D. M. HINGRAT. We assessed the inflammatory kinetic by performing plasma protein electrophoresis and haematology on blood samples drawn at intervals of 3.Peafowl. Haematology has been used in avian medicine for a far longer time. This surgery was considered to be the origin of an iatrogenic inflammatory condition. Y. Plasma protein electrophoresis and haematology thus both seem to be reliable laboratory examinations allowing early detection of acute inflammation conditions. SAINT JALME. France EARLY DETECTION OF AN ACUTE INFLAMMATORY CONDITION BY PLASMA PROTEIN ELECTROPHORESIS AND HAEMATOLOGY IN PEAFOWLS Y. Biol. ABSTRACT This study was conducted at Clères Zoological Park (France) where we followed the inflammatory kinetic after a superficial pectoralis tendonectomy on ten adult peafowl (Pavo cristatus). CHASTE . These examinations are both commonly recognized to be reliable diagnostic tools for many pathologic conditions in birds (WERNER 1999.Haematology . when serological tests fail to detect the agent of a disease. Agarose gel electrophoresis was performed with Hydragel protein 15/30 kits (Sebia. 6. we collected 1 ml of blood on EDTA. Plasma aliquots were loaded onto the gel and allowed to migrate for 8 min at 296 V (6mA) in a Sebia Hydrasis system. Samples were then thawed and re-homogenised 1 hour before analysis. Smears were made with the microscope slide and coverslip technique. 2. This surgery. no anti-inflammatory drugs were used postoperatively. 1994). HOCHLEITHNER. Since haemoglobin is known to migrate in the � fraction.TATUM 1998. � and �. Blood smears were made within 20 min after blood sampling to avoid artefacts caused by prolonged exposure to EDTA. We followed the inflammatory kinetic by performing plasma protein electrophoresis and haematology on blood samples drawn at the right jugular vein at intervals of 3. All surgeries went well and all the birds were released back into the park after the last blood sample was taken. The purpose of the study reported here was to determine the period of time between the beginning of an inflammatory condition and its detection by haematology and plasma protein electrophoresis. This kind of missing information often makes it difficult to be confident about the interpretation of the results and is therefore responsible for it’s underused in avian medicine. since it’s commonly accepted that plasma is preferable to serum. However. For haematology. and 291 . MATERIAL AND METHODS This study was conducted at the Clères zoological park (France) on ten healthy adult peafowl (Pavo cristatus) which were prevented to fly by superficial pectoralis tendonectomy (OLSEN. �2. �3. 28 and 52 hours after surgery. Total protein concentration was determined by the biuret method with the Roche integra system. performed under sterile conditions was considered to be the origin of an iatrogenic inflammatory condition. Once dried and coloured with amidoblack in the same system. France). gels were scanned by a high resolution Epson expression 1680 pro flat scanner. Albumin concentration includes prealbumin fraction. for protein electrophoresis on birds. For plasma protein electrophoresis. we collected 2 ml of blood on lithium heparin. BLANCO and HOFLE 2003). For the needs of this study. 1994) for legal and safety reasons. This study also aims at describing the kinetics of a normal inflammatory condition resulting from soft tissue surgery and to compare the efficiency of haematology versus plasma protein electrophoresis to diagnose such an acute inflammatory condition. Heparinised blood was immediately centrifuged and plasmas were stored in cryotubes for one week at -20°C (-4°F) to guarantee all reagents would be the same for the ulterior analysis. The electrophoretic curves and dosages of the different fractions were made with Phoresis Sebia’s software. Globulins were divided into 5 fractions: �1. 16. Protein electrophoresis were carried out on plasma. 11. as explained later in the discussion. care was taken to avoid haemolysis by taking blood samples with 20 G needles. because it is less prone to haemolysis and contain fibrinogen which is a characteristic acute phase protein (CRAY and TATUM 1998. we lack information about the delay between the beginning of an inflammatory process and the changes in the haemogram or in the proteinogram. 022). monocytes and immature WBC.dried immediately with a hair drier. France). TWBCs were performed within 5 hours after blood sampling. lymphocytes. The �2 fraction concentration then significantly decreased until T11 (Wilcoxon’s test: p<0.07) and T11 (Wilcoxon’s test: p<0.003). They all dramatically decreased between T0 and T3 (Wilcoxon’s test: p<0. Systat 7. Prealbumin values were very low and were included to albumin values.05). 3. but we found 3 fractions in the � region. Albumin and � globulin concentration slowly decreased from T3 to T16. Considering the small size of the population studied. The � fraction values then significantly increased from T11 to T52 (Wilcoxon’s test: p<0. The � fraction values significantly decreased between T6 and T11 and significantly increased between T16 and T28. �3) which varied independently. eosinophil granulocytes. 1. without any significant difference in the fraction rates and in albumin / globulin ratio between these two times. Total protein and protein fraction values all significantly varied with the time after surgery as illustrated in Fig. since looking similar to � and � peaks in other species. p=0.041) and then appeared to be stable. under phase contrast microscopy. and the eosinophil Unopette 5877 system (not available in France) by a 1/200 dilution of the blood with eosinophil dilution liquid containing B phloxine (LMR5004. as represented on Fig. but replacing the Neubauer haemocytometer by a Mallassez haemocytometer. whereas �1 and �3 fraction concentrations respectively increased after T16 (Wilcoxon’s test: p<0. and used to perform differential counts. using the indirect method described in avian haematology and cytology (CAMPBELL 1995). albumin concentration significantly increased between T16 and T28.001).07). RESULTS 3. Counts were made by counting 100 WBC which were distributed into 6 cell classes: heterophil granulocytes. It significantly decreased from T6 to T28 (Wilcoxon’s test: p<0. 1 Protein electrophoresis Peafowl electrophoretic patterns appeared to be divided into 7 fractions as illustrated in Fig. �2. Labo Moderne. The albumin / globulin ratio did not appear to significantly vary between T0 and T6. 2 (Friedman’s test: p<0. 3. Smears were coloured with the Wright and Giemsa coloration. From our interpretation of these fractions albumin showed the biggest peak. basophil granulocytes. we used Friedman’s two way analysis of variance and Wilcoxon’s signed rank test. Difference with T0 became significant after T16 (Wilcoxon’s test: Z=-2. All of them decreased between T0 and T3.981. The � fraction appeared to be divided into 3 sub-fractions (�1. After that.013).0 software (SPSS Inc 1997) was used for all analyses. The � and � globulin were identified on the electrophoretic curves. 292 . In this study. In peafowl. These results agree with what is usually described in the literature about acute inflammatory condition (CRAY and TATUM 1998). much data available in the literature was extrapolated from mammal serum protein electrophoresis. p=0. Unfortunately.022) and decreased again between T11 and T16 (Wilcoxon’s test: Z=-2.005).4). Further studies should be made to determine a standardised method of defining protein fractions on bird electrophoretic patterns. p=0. T3 and T6. we found that total protein and protein fraction concentrations all significantly decreased within 3 hours after surgery. 4. the publication of reference values is completely useless for the bird practitioner. basophil granulocyte.803. p=0. heterophil granulocyte and lymphocyte counts significantly varied with the time after surgery (Friedman’s test: p<0. Since there were no significant differences between the different fraction rates at T0. Without these precautions. Lymphocyte counts appeared to significantly decrease between T0 and T6 (Wilcoxon’s test: p<0. Significant increases were recorded after T16 (Wilcoxon’s test: p<0.037). Without the massive protein leakage occurring within the first 6 hours. � and � fraction values increased until the end of the study at T52. Eosinophil granulocyte. we can state that this protein leakage does not specifically affect one kind of protein. The � and � fractions contain acute phase proteins which rise during the first hours after an acute inflammatory condition like surgery. like it should in an active inflammatory condition (exudate).008) (Fig. and the inevitable 293 .293. 1999). It stresses the importance of explaining in each study how the author defined protein fraction. like massive blood loss or transudate occurring per and immediately postoperatively. The main acute phase protein in the � peak is generally accepted to be fibrinogen. which are well described in the literature. DISCUSSION This study highlights the problems encountered when dealing with electrophoretic patterns of bird Taxa other than raptors or psittacine birds.028). 2 Haematology TWBCs.3. T3 and T6 and between albumin / globulin ratios at T0. None of the variations recorded later appeared to be significant. This protein loss may be a passive phenomenon. only �1 and �3 fractions seem to contain acute phase proteins and may therefore be of diagnostic interest in an acute inflammatory condition.028). However � sub-fractions appeared to react in different ways. They significantly increased between T6 and T11 (Wilcoxon’s test: Z=2. Heterophil granulocyte counts dramatically increased between T0 and T3 (Wilcoxon’s test: Z=2. Another hypothesis could be a response to isoflurane anaesthesia (DRESSEN et al.191. After the decrease occurring in the first 3 hours. monocyte and immature cell counts did not significantly vary with the time after surgery. Figure 1 : example of a peafowl electrophoretic pattern at T0 Figure 2 : variation of protein fractions concentrations (g/dl) with the time after surgery (h) Asterisks represent significant differences between two successive measures (Wilcoxon’s test: p<0.05). 294 . Figure 4: variation of WBC counts (cell/µl) with the time after surgery (h) Asterisks represent significant differences between two successive counts (Wilcoxon’s test: p<0.05). 295 .Figure 3: variation of � fractions concentrations (g/dl) with the time after surgery (h) Asterisks represent significant differences between two successive measures (Wilcoxon’s test: p<0.05). haematology generally requires very experienced interpreters while plasma protein electrophoresis seems to be less prone to subjectivity. In return. Albumin / Globulin ratio decreases during an inflammatory condition. with chronic antigenic stimulation (CAMPBELL 1995). Monocyte counts appeared to be stable from the beginning to the end of the study. Without standards for defining each protein fraction. It was not therefore surprising that no significant variation of lymphocyte counts were recorded from the beginning to the end of this study. 1999). The rise in the heterophil counts is therefore undoubtedly more rapid than protein in answer to the acute inflammatory condition. However. haematology being even far more precocious. making plasma protein electrophoresis being reliable for early detection of an acute inflammatory condition in birds. protein electrophoresis in birds 296 . Haematology and plasma protein electrophoresis are both reliable laboratory tests to diagnose acute inflammatory conditions within the first 16 hours. However. the rise of acute phase protein concentrations should have probably been far more precocious and massive than demonstrated in this study. since A/ G ratio was stable within the first 11 hours even though total protein was decreased. In avian medicine. because many immature cells were found in the blood stream in response to the massive consumption of granulocytes at the inflammation site. Further studies concluded that plasma protein electrophoresis was useful for acute diagnostic of chlamydophilosis (< 2 weeks). Monocyte counts only rise in chronic pathological conditions in which chemotactic agents are released (like aspergillosis. the most relevant criteria allowing the assessment of the bird’s health status seems to be the albumin / globulin ratio. Haematology appeared to be difficult to interpret in our case. This study clearly demonstrates that A/G ratio decreases and acute phase proteins rise at least within 16 hours after the beginning of an inflammatory condition. The dramatically increase of heterophil counts within the first 3 hours postoperatively was probably due to massive endogen corticosteroid discharge in response to the surgical trauma (CAMPBELL 1995). Lymphocyte counts are generally thought to increase in chronic diseases. chlamydophilosis. before seroconversion of the avian patient (CRAY and TATUM. the decrease could be related to the haemodilution due to frequent blood samples. In this study. This study highlights the fact that A/G ratio values must be interpreted in the light of total protein concentration. in accordance with what is generally accepted. as other authors reported in previous publications (LUMEIJ 1997). 1998). the successive decrease and increase of these lymphocyte counts within 16 hours after surgery is quite difficult to understand and should be investigated in further studies. avian tuberculosis or massive tissue necrosis) (CAMPBELL 1995). due to increased globulin fractions and decreased albumin fraction (mainly in chronic inflammatory conditions). The slow decrease of � fraction values is normal because this fraction is commonly known to contain immunoglobulin in birds (CRAY and TATUM 1998) and is therefore supposed only to increase in chronic inflammatory conditions (>10 days).Dilution artefact related to successive blood samples (DRESSEN et al. underlining the fact that protein electrophoresis may be a valuable laboratory test to diagnose acute inflammatory conditions. protein electrophoresis was first recognised for the detection of chronic subclinical inflammatory conditions such as aspergillosis. at the present time. tuberculosis and psittacosis as well as egg peritonitis (MARGOLIN 1995). 4. 5. Plasma protein electrophoresis as a diagnostic and prognostic tool in raptors. 12: 4 . France Email: yannick.10. 8. In: RITCHIE B W. San Diego: Academic Press 1997. Avian clinical biochemistry. 859 . 3. OLSEN J H. CRAY C and TATUM L. BLANCO J M and HOFLE U. J Avian Med Surg 1998.662. HARVEY J W and BRUSS M L (eds): Clinical biochemistry of domestic animals. 2. (ed): Avian medicine: principles and application. Muséum national d’Histoire naturelle. In: RITCHIE B W.1272. Lake Worth: Wingers Publishing Inc 1994. WIMSATT J and BURKHARD M J. MARGOLIN T. Avian haematology and cytology second edition.29. Proc Assoc Avian Vet. The diagnostic utility of serum protein electrophoresis. WERNER L L and REAVILL D R. 1270 . or in the choice of the equipment. 76690 Clères.179. 6. to A SADRIN. Roman. F BURON for their English language proof-reading of this article. In: KANEKO J J. HARRISSON G J and HARRISSON L R (eds): Avian medicine: principles and application. Veterinary Clinics of North America: Exotic Animal Practice 1999. The effects of isoflurane anesthesia on hematologic and plasma biochemical values of American kestrels (Falco sparverius). Lake Worth: Wingers publishing 1994. AUTHORS ADRESS Y. Normal electrophoretic values in cockatiels (Nymphicus hollandicus) and factors affecting these values. 237 . MsC. DRESSEN PJ. CAMPBELL T W. Proc Eur Assoc Avian Vet.238. Anseriformes. 3 . Biochemistries.66. DVM. J Avian Med Surg 1999. 32 avenue du parc. Iowa: Iowa State University Press/Ames 1995. the technique or the way of defining protein fractions. Nashville 1995: 65 . LUMEIJ J T. 5 AKNOWLEDGMENTS To Sebia France for their generous gift of Hydragel protein 15/30 kits. 7. 6 CITATION INDEX 1. Tenerife 2003: 256 .roman@online. 13(3): 173 . C HOOTS. 2: 651 .appears to be far less standardised than haematology and results clearly depend on the choice of serum instead of plasma. Ames. Application of protein electrophoresis in avian diagnostic testing. HOCHLEITHNER M.261.861. HARRISSON G H and HARRISSON L R.fr 297 . D VIDAL. 9. Le Parc de Clères Jean Delacour. habil. med. In birds 298 . i.Confocal scanning technique . availability for combination with FAG and slit lamp examination and thus availability for daily routine follow ups of ophthalmological treatment procedures. vet. i. that using an infrared device as an observation light allows documentation in pupil diameters as small as 4 mm. vet. Prof. med. Cert Vet Ophthalmol KEYWORDS Digital scanning ophthalmoscope . even in non-mydriatic pupils. that the new digital scanning ophthalmoscope includes major advances for avian ophthalmology such as short examination periods avoiding stress situations due to real-time image capture.Fluorescence angiography . the fovea. Dr. Using ophthalmoscopy besides the vitreous the – in birds anangiotic – retina. Dr. Dip ECAMS. Ludwig-Maximilians-University Munich. e.Individual identification ABSTRACT Ophthalmoscopy is a routine ophthalmological procedure for examining ocular structures located behind the lens.. rapid availability of single and sequence still images as well as video documentations. Switzerland) which allows rapid digital documentation of fundus.Fundus oculi . Korbel. T. Germany DIGITAL SCANNING OPHTHALMOSCOPY IN BIRDS R. Wild Medtec. choroid and sclera) may be seen. The present study is focussing on ophthalmoscopy using a new scanning digital ophthalmoscope (SDO-A. Zürich. Depending on the pigmentation of the retinal pigment epithelium (RPE) the underlying vascular pattern of the choroid and – most often due to minimal or absent pigmentation of the RPE – in nocturnal species the sclera as the outer layer of the three ocular layers (retina. the vitreous and the fundus oculi.Clinic for Birds. e. the pecten and at its basis the more or less obscured optic nerve head may be visualized. 1 INTRODUCTION Ophthalmoscopy is a routine ophthalmological procedure for examining ocular structures located behind the lens. Investigation results in various raptor and psittacine species as well as pigeons show. Furthermore results show. Depending on the anatomy additionally structures like the ora serata. by performing paracentesis. Zürich. structures of the lens including associated structures like the processus ciliares and even anterior eye chamber structures like the filtration angle with the pectinate ligamentum. Zürich. most often arising from the pecten oculi. Additionally. Whereas for birds with a pupil diameter of less than 6 mm the special laboratory animal fundus cameras. For evaluation purposes this camera was used taking stills and videos in healthy diurnal raptors including common buzzards (Buteo buteo. Wild Medtec. iridal structures and more may be examined using an ophthalmoscope. nocturnal raptors including great horned owls (Bubo bubo. n = 4). whereas for avian ophthalmology and birds with a pupil diameter of more than 6 mm (most raptors. enabling the examiner not only to take still photos but also videos. large psittacines) handheld systems like the KOWA RC 2 or KOWA Genesis for Animals and KOWAS Genesesis are suitable devices. psittacines. as technique to diagnose fundus disorders associated to the vascular pattern. while more than 35 % of all traumatised birds may develop intraocular haemorrhages. as these structures are like a fingerprint and may be used as a fraud proof method for individual markers without using rings or micro-transponders. various raptor and psittacine species (total: n = 18) with diseased eyes were examined. the left pupil was dilated using intracameral application of a 0. digital cameras have been made available. n = 4). the pars plicata of the retina. n = 12). Switzerland) was used. like the KOWA Genesis for laboratory animals have to be used. Up to the most recent past. Switzerland) which allows rapid digital documentation of the fundus. These post-traumatic damages have to be diagnosed rapidly followed by effective therapy as visual impairment and blindness may be a possible result. Furthermore visualising fundus structures will be of future importance for individual identification by pictorial documentation of individual vascular patterns and pecten structures. 2000. n = 4) and racing pigeons (Columba livia. including blue fronted amazons (Amazona aestiva.03 cc) depending on the size of the anterior eye chamber. KORBEL 2000). Most recently. For examination purposes of each individual. Wild Medtec. 2 MATERIAL AND METHODS A scanning digital ophthalmoscope (SDO-A. n = 12).01 – 0. allowing only steady images including serial image documentation (for fluorescence angiography) with up to 1 frames per second lacking the possibility for taking clips or movies. only analogous fundus camera systems (cameras using slide films) were available. the so-called fundus cameras may be used.3 % d-tubocurine solution (0. The pupil diameter of these birds examined covered a range down to 4 mm in diameter. Sophisticated diagnostic techniques based on ophthalmoscopy include fluorescein angiography.ophthalmoscopy is indispensable in traumatised birds. 299 . n = 12). One of these systems is the newly available scanning digital ophthalmoscope (SDO-A. For image documentation. while the right pupil was left un-dilated (KORBEL et al. tawny owls (Strix aluco. macaws (Ara macao. (eds): Raptor Biomedicine III. while combined with the ease of handheld and mobile operation procedure. med. Germany Email: korbel@lmu. even in non-mydriatic pupils in raptors. Dr. D – 85764 Oberschleissheim.3 RESULTS Investigation results in various raptor and psittacine species as well as pigeons showed. the digital camera may serve as an aid for identification of individuals by documenting the vascular pattern of the choroid rather as an alternative to rings or micro-transponders. availability for combination with FAG and slit lamp examination and thus availability for daily routine follow ups of ophthalmological treatment procedures by documenting anterior as well as posterior eye segment structures.de 300 . Dr. Ludwig-Maximilians-Universität München. et al. that using the new digital fundus camera allowed high resolution black and white as well as colour images. REDIG PT. Video fluorescein angiography in the eyes of various raptors and mammals. et al. e. Sonnenstrasse 18. vet habil. Able to document anterior as well as posterior fundus segments in all birds with a pupil diameter down to 4 mm. AUTHORS ADRESS R. KORBEL R. Results have been documented producing jpeg and tiff files and will be documented within the presentation. vet. Klinik für Vögel. Korbel.95. rapid availability of single and sequence still images as well as video documentations. real-time capture and video tracking.. Dip ECAMS. med. Lake Worth: Zool Educ Network 2000: 179 – 94. 2.g. 4 DISCUSSION The results showed that the new digital scanning ophthalmoscope includes major advances for avian ophthalmology such as short examination periods avoiding stress situations due to real-time image capture. REMPLE JD. Proc Assoc Avian Vet. Portland 2000: 89 . 5 CITATION INDEX 1. In: LUMEIJ JT. Prof. REDIG PT. Additionally. T. Diseases of the posterior eye segment. NELL B. KORBEL RT. calcium deficiency. E and selenium deficiency abdominal hernia. there is still a gap between knowledge and understanding of reproductive disorders in avian species and mammals (CROSTA et al. in which the different imaging techniques (endoscopy.Radiology . radiographs and ultrasounds). infectious diseases of the oviduct and genetic predisposition (CROSTA 2000. torsion of the uterus. GERLACH 1998. The paper presents common and unusual avian obstetrics cases. CROSTA et al. excessive egg laying. obesity. 1 INTRODUCTION Although disorders of the reproductive apparatus represent an important part of domestic animal medicine. thin eggshell. neoplasia. and/or useful for the resolution of the case. Vitamin A. 2003). Causes of egg retention are multifactorial and involve: oversized or misshapen eggs. 2003). dysfunction of the oviduct. 2003).Obstetrics . Timossi. Dystocia can be described as the mechanical obstruction of the egg passage in the caudal part of the oviduct (uterus and vagina) (GERLACH 1998).Ultrasound ABSTRACT Obstetrics is a relatively unexplored field of avian medicine and radiology is still the most used diagnostic tool in birds. but still the diagnosis and subsequent treatment protocol of several female bird conditions are not well established. Crosta.Endoscopy . Egg retention and dystocia are among the most frequent obstetric disorders of birds (CROSTA et al. DVM KEYWORDS Birds .Loro Parque Tenerife – Canary Islands. have been effective in reaching a diagnosis. Some specific diseases of the female and male reproductive system have been described (BOWLES 2002. DVM and L. 1997) 301 . Spain IMAGING TECHNIQUES IN AVIAN OBSTETRICS L. They are also the most frequent avian emergencies. OROSZ et al. Egg retention may be described as an egg which is not laid within the normal time (CROSTA 2000). and it was confirmed radiologically. In some cases. To prevent subsequent egg laying. The shell fragments should pass within a few days. 2003). SQ fluids. The presence of an egg should be ascertained by radiographs or ultrasound. leg paralysis or respiratory difficulty (OROSZ et al. the egg will pass without further treatment (JOYNER 1994). In minimally depressed birds. but the cloaca was emptied and lubricated. at the utero-vaginal sphincter level (GERLACH 1998). 2 CASE REPORTS Three example cases are taken into consideration for this study. Further. 1992). Therapy depends on the physical status of the patient and the severity of the clinical signs (OROSZ et al. 1997). or there were soft shelled eggs. physical examination. At clinical examination a suspect diagnosis of egg retention was made. Case No 1 A 3 years old female emerald-collared parakeet (Psittacula calthorpae) was found resting in the bottom of her cage. a PGE-2 gel was inserted 302 . megestrol acetate can be given (CROSTA et al. more aggressive therapy should be considered. Other signs might include lameness. Also. Since there were several differences. either because the birds belonged to different species. 1997). due to the patient conditions. vitamin E/selenium. blood samples were not collected. which is extremely risky for the recently egg-bound hen. radiography and/or ultrasonography. with distended coelom and tenesmus. ultrasound is the preferred diagnostic tool (CROSTA 2000). fluids. 2004) and also several improvements have been done using ultrasounds to scan the avian female reproductive system. calcium and antibiotics. antibiotics and diluted calcium gluconate were administered. and run the risk of kidney damage and intestinal impaction. different techniques had to be used for the diagnosis and for the resolution of the problem. 2003). through the cloaca or by transcoelomic access (OROSZ et al. After supportive therapy with fluids. or the removal through the cloaca is not possible. In birds which are not able to pass faeces and urine. The birds should be placed in a warmed and humidified cage (CROSTA et al. The egg was considered too large to be easily delivered and the bird was extremely weak and judged at risk for any invasive treatment. Vitamin D and calcium gluconate (HARRISON et al. Every case is involving a female bird that became egg bound.The diagnosis of egg retention should be based on clinical signs. as coelomic palpation is not sensitive enough. In the case of egg retention with a soft egg with non-calcified egg. such as eggshell fragments are not delivered. Suspicion should arise if the clinician is presented with a female patient. or if the adhesions between the egg and the uterus are severe. Patients which are still able to pass faeces and urine should be given antibiotics. To promote oviduct contractions Prostaglandin E2-gel can be inserted in the cloaca. Recently some publications have appeared about endoscopy of the oviduct (CROSTA et al. or even because more than one egg was involved in the problem. a coeliotomy might be necessary (JOYNER 1994). 1997). ovocentesis should be performed. giving the impression of a relatively soft-shelled egg. most important. and the bird’s condition was clearly worsening: patient was not eating. The endoscope. and a transabdominal ovocentesis was also to be excluded. The parakeet was quickly anaesthetized with isoflorane. Furthermore. The cloaca was carefully emptied. Since this was not possible. to a point where it could be visualized without the endoscope. in a more distal position. and eventually be ready to perform a hysterotomy to retrieve the collapsed eggshell. After 12 hours. the eggshell collapsed with no need for a manual compression. The parakeet recovered uneventfully from the surgery. Unfortunately the 5-Fr grasping forceps were too small for a good grip. and a 18-G needle. but. was presented for abdominal distension that was lasting since several days. but there was no sign of egg laying.into the cloaca. was done. the egg had not been delivered and the patient’s conditions were worsening again. but simply opening the cloaca with a small speculum. even if this was not apparent at x-ray. since it was soft shelled. because one egg was very difficult to locate. together with its examination sheath. was carefully introduced into the distal oviduct. 303 . was apparently poorly calcified. and only small amounts of eggshell were retrieved in this way. The bird was then put in warmed cage and food and water were provided. The cloaca was clearly inflamed. passing through the abdominal and oviduct walls. Whole body X-rays were taken as well as blood for haematology and blood chemistry and microbiological samples. The following day the bird received enrofloxacin and meloxicam PO. using gently manoeuvred cotton swabs. and a cloacoscopy. connected to a 5ml syringe was inserted into the egg. Case No 2 A 9 years old female gentoo penguin (Pygoscelis papua). The endoscope was retrieved and inserted into an operating sheath with an instrument channel and the whole system was inserted again into the cloaca. After several days the bird was still doing well. It was thus decided to have a thoroughly endoscopic examination of the cloaca. and an attempt to visualize the eggshell through the left oviduct-cloacal sphincter. The cloaca was carefully emptied again. nor urine were passed. for the presence of the oversized and retained egg. but the oviduct-cloacal sphincter could easily be located. one of which. it was decided to perform a trans-abdominal ovocentesis: the abdomen was cleaned and disinfected. and the collapsed eggshell could be visualized. Nevertheless it was possible to pull the eggshell further distally through the oviduct. During the aspiration of the egg content. Small curved mosquito forceps were then inserted and the eggshell could be firmly grasped and fully retrieved. no faeces. was contraindicated. The bird was anaesthetized again and entubated. by face mask. This was likely due to the relief of the intra-abdominal pressure. was started. X-ray was pointing out the presence of two eggs. PGE-2). the egg had not been delivered yet. oral megestrole acetate was added to treatment. 6 hours after the procedure. On the other hand the use of drug that may induce the contraction of the oviduct (oxytocin. The eggshell was located again and it was grasped with the small flexible 5-Fr grasping forceps. Immediately after recovering from the minor procedure the bird looked better. but since the clinical situation was not evolving. The patient. She was treated for few days after surgery. The oviduct was fixed with two stay sutures and the egg content was gently aspirated with a 20ml syringe. The patient overcame the procedure with no apparent problems. For this reason leuprolide acetate was administered and repeated at day 15 and 30. After one week ultrasounds showed an oviduct that was still dilated. Once the egg was emptied. Also Meloxicam was administered for a week. but it was apparently non effective. The penguin recovered uneventfully from the surgery. The bird was given Calcium and vitamins and. and the bird was not allowed to swim for 10 days after surgery. The patient was anaesthetised and maintained with isoflorane and the oviduct was easily identified after entering the coeloma. Exactly one year after surgery. Case No 3 A female blue and gold macaw (Ara ararauna). X-ray revealed the presence of a large egg. for rescuing dystocial eggs has been described (JENKINS 2000. Oviduct and abdominal wall were sutured as described in other avian species. but it could be manually extracted. The egg was then laid after 30 minutes. 304 . she laid a normal egg. Case No 2. Case No 3. a macaw used for a show. The second egg was in a very proximal position. To facilitate this manoeuvre. the egg content was aspirated by guiding the needle with the help of ultrasounds. was presented for abdominal distension lasting several days. so that the egg would naturally drop more distal in the coeloma. The egg wasn’t laid after this time and PGE-2 were administered intracloacally. was reincorporated into the program and has been displaying during the show since then. The bird was doing fine immediately after the procedure. A treatment with megestrole was started. so it could not be visualized. The bird was monitored for few days. A long term course of antibiotics and megestrole were used for the penguin. The day after ultrasounds ascertained the presence of a large soft shelled egg. ECHOLS 2002). As soon as the oviduct was opened the soft shelled egg was spotted. and is still with her cage mate in a large aviary where we are waiting for her to breed. it was decided to wait 24 hours. a helper was needed to hold the penguin in a 45º position. The patient has been fine since then. but it has never been published in penguins. when the wound was healed and the bird was returned to the exhibit. A laparotomy technique. it was retrieved with a small forceps.It was decided to have the eggs out surgically. and has been in very good clinical conditions since then. but a new treatment was needed to have a complete recovery. since its conditions were good. 3 RESULTS Case No 1. to limit possible postoperative infections and help reducing temporarily the size and inflammation of the oviduct. to date. 305 . but does not come out in a natural way. Finally. still it was too far in the long oviduct of penguins for the endoscope. Also. size and age of the patient. probably because of the feathering of penguins that is too thick and ultrasound window that is too small. Case No 1 is a good example of how endoscopy has few limits in birds: once an egg has been imploded. The authors want to stress the need of a multidisciplinary approach to the diagnosis of the diseases of the avian female reproductive tract. number. Case No 3 is a clear example of how ultrasounds are a great help first in the diagnosis. While X-ray remains the main imaging technique in avian obstetrics. It is obvious that these are simple examples of a very complicated field.4 DISCUSSION Obstetrics is still a relatively unexplored field of avian medicine and radiology has historically been the most used diagnostic tool in birds. when a hen is egg-bound with a soft shelled egg. in which many different presentations (species. This especially because normally calcified eggs are well radio-opaque. quality and relative size of the eggs) can lead the clinician to prefer one technique instead of another. but the access to good quality tools and a creative mind may lead to discoveries also in this field. one soft-shelled egg and a normally calcified one. endoscopy and ultrasonography. but radiology is still a great diagnostic help. the relative large size of the avian egg may limit the application of endoscopy in avian obstetrics. and then in the resolution of unusual obstetric cases. the endoscope was not able to reach the soft shelled egg. This likely because entrance to the oviduct is not easy to identify and special skills and very good instruments are needed to access this organ. Ultrasounds could not spot the eggs. So the only option was to go for a standard surgery and rescue the eggs. This is because. there are cases in which endoscopy and ultrasound do not find a good application. Experience and constant practice are needed to become proficient in the use of sophisticated techniques. This bird has been diagnosed to be egg-bound with. even if the egg was very low in the abdomen. but its application to obstetrics has been very limited. but the authors showed that they have an application also in the treatment of egg-binding. Endoscopy is one of the most used techniques in bird medicine. such as radiology. this paper shows how some of the techniques widely used in other animals are very promising even in birds. The gentoo penguin in Case No 2 is giving an example. at least. On the other hand. Ultrasounds have been already advocated as the only diagnostic tool able to confirm the presence of non-calcified eggs. the use of the endoscope may solve the problem in a relatively non-traumatic way. Stuttgart.5 CITATION INDEX 1. Urogenital disorders. ECHOLS SM: Surgery of the Avian Reproductive Tract. 2000. diagnosis and diseases of the avian reproductive tract. Cremona. 6(1): 10 . J Assoc Avian Vet 1992. 748 .692.644. HARRISON GJ and HARRISON LR (eds. Lake Worth: Wingers Publishing 1994.804. Philadelphia: WB Saunders.15. 7.): Avian Medicine: Principles and Application. HARRISON G. Loro Parque s/n 38400 Puerto de la Cruz Tenerife. 196/o-196/za.com 306 . Italy. 9. In: Wiesner E (Hrsg. Seminars in Avian and Exotic Pet Medicine 2002: 11(4): 177 . Exotic DVM 2004. BÜRKLE M and TIMOSSI L: Physiology. BÜRKLE M and TIMOSSI L: Endoscopy-assisted Resolution of Egg Binding in an Emerald-collared Parakeet. 3.1): 19 .195.): Handlexikon der Tierärztlichen Praxis. GERLACH H. et al: Roundtable discussion: Emergency medicine. 6. In: Proc Corso SIVAE (Società Italiana Veterinari per Animali Esotici) di Medicina e Chirurgia Aviare. 1997. 614 . Erkrankungen. (3:3): 637 .83. Vet Clin North Am: Exotic Animal Practice 2003. Spain Email: lorenzo_birdvet@yahoo. 5. DORRESTEIN GM. et al. 2. BOWLES HL: Reproductive Diseases of Pet Bird Species.22. 6(3): 57 . DORRESTEIN GM and SPEER BL. ( 6. 1998. AUTHORS ADRESS Lorenzo CROSTA DVM Veterinary Director Loro Parque & Loro Parque Foundation Avda. OROSZ SE. Vet Clin North Am: Exotic Animal Practice 2002. GERLACH H. CROSTA L: Patologie della Riproduzione e dell’Apparato Riproduttore. Ferdinand Enke. In: RITCHIE BR.506. 5(2): 489 . In: ALTMAN RB. Eibildende Organe. CROSTA L. (eds): Avian Medicine and Surgery. JENKINS JR: Surgery of the Avian Reproductive and Gastrointestinal Systems. 8. CLUBB SL. Vet Clin North Am: Exotic Animal Practice 2000. 10. 4. JOYNER KL: Theriogenology. CROSTA L. Hässig. Hatt. 1998) and has been adapted to measure bone quality in vivo in poultry (KORVER et al. M. To our knowledge QCT has not been described in psittacine birds.. Peripheral quantitative computer tomography (pQCT) is a non-invasive technique commonly used to measure bone mineral content (BMC) and bone mineral density (BMD) in vivo. 307 . Dr. Switzerland QUANTITATIVE COMPUTED TOMOGRAPHY FOR THE ASSESSMENT OF BONE DENSITY IN BUDGERIGARS : A PILOT STUDY I. vet. 2004). 2004. These results demonstrate that pQCT may be used for in vivo to evaluate bone quality in budgerigars. BMC and BMD values were higher in the female group compared to the male group. We evaluated 10 adult budgerigars (Melopsittacus undulatus) (5 females and 5 males) housed under identical conditions and identical feeding regime for BMC and BMD. Dr. A. med. Vetsuisse Faculty.-M. Dr. 2004).Budgerigar ABSTRACT Peripheral quantitative computer tomography (pQCT) is a non-invasive technique commonly used to measure bone mineral content (BMC) and bone mineral density (BMD). 1 INTRODUCTION The bone parameters commonly used to investigate bone mineral quality are bone ash. vet. vet KEYWORDS Computed tomography . LIESEGANG et al. PQCT is used diagnostically in humans to assess osteoporosis (FORMICA et al. The main disadvantage of these methods is that they are invasive. J. Liesegang.. med. vet. med.Division of Zoo Animals and Exotic Pets. Fischer. The measurements were carried out on the tibiotarsus with peripheral quantitative computer tomography under general anaesthesia.Bone mineral density . med. bone breaking strength and atomic absorption spectrophotometry (KORVER et al. Dr. University of Zurich.. Prof.Bone mineral content . PQCT is used diagnostically in humans and has been adapted to measure bone quality in vivo in poultry. 1 389. Germany).2 BMC mg/cm 1. Mean values of BMD (xfemale = 509.04 mg/cm) were higher in the female compared to the male group. xmale = 373. Switzerland) plus carrots and a mineral supplement (Quikon.5 335.3 414.44 1.3 444. M= male.2 MATERIAL AND METHODS We evaluated 10 adult budgerigars (Melopsittacus undulatus) (5 females and 5 males) housed under identical conditions and identical feeding regime.95 1.9 330. xmale = 1. Statistical data were analyzed using ANOVA (Systat.23 0.86 308 . The budgerigars were anaesthetized with isoflurane and positioned in dorsal recumbency with wings in normal position. 3 RESULTS Bone mineral density and bone mineral content were evaluated in the tibiotarsus of a budgerigar with three measurements.0001) and BMD (P = 0. Stratec Medizinaltechnik GmbH. Jordi AG. SWV English budgerigar-food. The values of BMD and BMC were similar within the male or female group. Germany).93 1. respectively.9 713. Table 1 Bone mineral density and bone mineral content of the tibiotarsus of budgerigars evaluated with peripheral quantitative computed tomography.65 mg/cm . Bocholt.94 1.04 1. F= female N° 1 2 3 4 5 6 7 8 9 10 Sex M M M M M F F F F F BMD mg/cm3 427.66 mg/cm3) and BMC (xfemale = 1. Significant differences were seen in BMC (p<0. IL). The right leg was fixated in extended position.Signau.55 2.49 0. The budgerigars received a commercial seed mixture (90 Hammer.4 382. Firma Quiko.0003) between males and females.03 0.3 594. The values were reproducible. Pforzheim. Total bone mineral density (BMD) and content (BMC) were measured in the right tibitotarsus with peripheral quantitative computer tomography (pQCT) (Stratec XCT 2000 bone scanner.82 mg/cm3 .5 391. ch 309 . 2004: 220 . University of Zurich. Before appliance in other psittacine birds the pQCT has to be evaluated. to evaluate the calcium metabolism or to control fracture healing. KORVER D. 3. These results demonstrate that pQCT may be used for in vivo to evaluate bone quality in budgerigars. 8: 460 . 2. The difference between male and female group may be due to calcium storage in the bones for eggshell formation. AUTHORS ADDRESS Isabelle Fischer.467. Einfluss von durch Hitzebehandlung des Futters reduzierter Phytaseaktivität auf die Phosphorversorgung und Knochenparameter von Broilern. Vetsuisse Faculty. 83: 222 . CROSMAN F.229. Schriftenreihe aus dem Institut für Nutztierwissenschaften. med. LIESEGANG A. 8057 Zurich. Switzerland Email: ifischer@vetclinics. Assessing bone mineral density in vivo: quantitative computed tomography. MESSIKOMMER R and WENK C.unizh. vet. 5 CITATION INDEX 1. Winterthurerstrasse 260. Comparative assessment of bone mineral measurements using dual x-ray absorptiometry and peripheral quantitative computed tomography. Division of Zoo Animals and Exotic Pets. We encourage the evaluation of pQCT to investigate the influence of different diets on bone quality. SAUNDERS-BLADE J and NADEAU K. Dr. Tagungsbericht. et al.223 FORMICA C. ETH Zürich. J.4 DISCUSSION The BMD and BMC values of this study show a similar distribution within the same gender group and we were able to reproduce the values with several measurements in the same bird. Poult Sci 2004. NIEVES. Osteoporos Int 1998. serves to remind all those charged with maintaining hospital facilities. all patients and the biosecurity status of the facility. Forbes BVetMed CBiol MIBiol DipECAMS FRCVS S. Swindon United Kingdom BIOSECURITY OF THE AVIAN HOSPITAL FACILITY. which aims to minimise surfaces which may facilitate pathogen build up. whilst Ward B was disinfected by fogging. KEYWORDS Pathogens – Hospital – Biosecurity – Fogging . 1 INTRODUCTION Avian and Exotic Animal Clinicians are faced with seriously pathogenic and on occasions fastidious pathogens on a daily basis. In the human medical field where design and material cost limitations may be less restrictive. designs in surfaces which are more readily cleaned and disinfected are well documented and will not be discussed here. Ward A was disinfected in a traditional manner for the first week. BY DESIGN. PROTOCOLS AND PROCEDURES N. The results will be presented and discussed at the conference and will be published in the near future elsewhere.Great Western Referrals. The principles of modern hospital design and standards.Disinfection ABSTRACT Environmental sampling was conducted in two equally busy wards dealing with avian patients. ‘Surface samples’ and ‘settle plates’ were employed to evaluate the levels of surface and air borne contamination. In the third week. These pathogens present a risk to staff. of the critical importance of optimal biosecurity procedures. In the second week. both wards were disinfected in a traditional manner. following conventional surface disinfection as opposed to F10 fogging. which has led to a significant number of deaths and endangered many thousands of other patients. In the third week. Ward A was disinfected by fogging. Microbiology samples were taken on weekdays of each week. whilst Ward B was disinfected in the traditional manner. Smith BVetMed MRCVS. Ward A was disinfected by fogging. whilst Ward B was disinfected in the traditional manner. 310 . the recent dramatic increase in methicillin-resistant Staphylococcus aureus (MRSA). Such areas tend to be busy and are likely to contain significant amounts of equipment. The active constituents of F10 Super Concentrate Disinfectant (“F10”) include quaternary ammonium and biguanide compounds. irregular hard surfaces is difficult and considered that the problem may be solved by using gaseous decontamination methods (FRENCH et al 2004). before and after traditional cleaning and disinfection procedures. Mycobacterium spp.. as this is a more controlled environment than a treatment or procedure area. whilst (Circovirus) is also extremely persistent in the environment.4%. BENDHEIM and DAVIDSON (2005) in a recently published survey of prevalence of circovirus in breeding establishments in Israel found a 20% level. the greatest risk is then posed by consulting areas as well as central procedure or treatment areas.. edetate and non-toxic 311 .e. and a cross over trial cold be readily performed. The latter method reduced the contamination level by 98. through which many different patients will pass in a day. after a patient has vacated an area). i. air borne. The aim of this trial was to compare the efficacy of two cleaning modalities. but those of greatest concern to the authors are Chlamydophila spp. In a typical avian practice it is likely that over 50% of these will appear ill. Within the ward cross contamination can be minimised by maintaining birds in their own ‘discrete individually air extracted space’. In effect traditional cleaning techniques reduced the MRSA levels by only 11%. soft furnishings.. documented MRSA levels of 74% and 66% respectively. by the use of the disinfectant F10 (STANFORD personal communication). with hydrogen peroxide vapour disinfection (Bioquell PLC). such that the clinician is not aware of a potential contamination risk. such that the risk of infection from one bird to another is minimised. These pathogens are highlighted as they are commonly present in apparently normal birds. The trial was to be conducted in the ‘common ward area’ of two wards (i. Within the avian and exotic animal setting.FRENCH et al (2004). Cryptosporidia. Pseudomonas spp. general ward. with the intention of then applying an improved protocol to all clinical areas. Circovirus and the causative agent of Psittacine Proventricular Dilation Syndrome.e. sampling a London Teaching hospital environment. Where ever possible equipment and disposables should be enclosed such that air borne contamination risk is minimised. is minimised. In this way the risk of contamination of them..e. Chlamydophila and Circovirus are typically spread in feather or faecal dust. Herpes virus. the clinician is commonly faced with a range of highly pathogenic organisms (Salmonella spp. excluding patient areas which benefit for ‘discrete individually air extracted space’). Overalls and shoes used in contaminated areas can be banned from the cleanest areas (theatres). Campylobacter spp. Circovirus has previously been shown to be controlled in an aviary situation. Those authors then compared these results collected after conventional terminal cleaning (i. whilst the incidence of Chlamydophila in similar situations has previously been reported to range from 10-60%. or from them respectively. whilst a further significant percentage will appear well but be carrying potentially contagious diseases. Hospitals should be designed with the very clean areas (theatres) and more contaminated areas (isolation ward. autopsy room) positioned at ‘dead-ends’. However it is recognised that the cleaning of complex environments containing medical equipment. by fogging application. However having taken these steps. floor wall interface. shelving at a height of 1 metre). Hampshire. Basingstoke. is said to be virucidal. at five sites (door furniture. Biguanide compounds and quaternary ammonium compounds act via interference with cytoplasmic membrane function. bactericidal. whilst Ward B was disinfected by fogging. UK). were moistened in sterile saline and used to sample pre determined surfaces of approximately 25cm2 by standardised swabbing in two directions at right angles. this was scored ‘2’. amongst other actions.1. Growth was recorded semi-quantitatively: If growth remained in the area of primary culture. following conventional surface disinfection as opposed to F10 fogging. In the third week. were placed in each ward on a daily basis. 2 MATERIALS AND METHODS This investigation was conducted in the ‘avian and exotic animal’ ward facility of the authors’ veterinary facility. 2. causing.ampholytic surfactants. following streaking out. on 5 consecutive days. swabs were re-suspended by agitation in 2ml of sterile saline for 30 seconds and ten left to stand for a further 5 minutes. patient cage fronts. Prior to plating the saline containing the swabs were re-agitated and the swabs were then used to inoculate Blood Agar and Maconkey Agar Plates (Oxoid. Hampshire. this scored ‘1’. Settle plates contained blood agar (BA) (Oxoid. quaternary ammonium compounds cause increased permeability of the outer member in Gram negative bacteria (MAILLARD 2002). Additionally. whilst if the growth extended beyond the area of initial inoculation. as a routine disinfectant procedure. wall. ‘Surface samples’ and ‘settle plates’ were employed to evaluate the levels of surface and air borne contamination. Ward A was disinfected by fogging. whilst Ward B was disinfected in the traditional manner. Environmental sampling was conducted in two equally busy wards dealing with avian patients. both wards were disinfected in a traditional manner. 12 hours after disinfection and left open for 60 minutes. After collection. Basingstoke. Samples were taken on weekdays of each week. Ward A was disinfected in a traditional manner for the first week. prior to the start of the next day. The authors wished to evaluate ‘fogged’ F10. coagulation of cytoplasmic constituents. fungicidal and sporicidal (VERWOERD 2001). EDTA disrupts cell wall integrity by chelating metal ions (FOSTER and DEBOER 1998). F10. with the exact formulary remaining a commercial secret. In the second week. Aerosol therapy with F10 has been successfully used in cases of lower respiratory tract infections at a dilution of 1:250 and is well tolerated by patients (CHITTY 2002). Environmental surface sampling and microbiological methods Cotton tipped sterile swabs. Cleaning was performed at the end of each working day and samples were collected some 12 hours later. The air conditioning system was inactivated between disinfection and the time when sampling was completed. Conventional Cleaning/ 312 . Minimal human and patient movements occurred in the area in the interim. compared to more traditional veterinary hospital cleaning methods (see below). which were then incubated at 37oC for 36 hours. UK). Disinfection: the conventional cleaning technique used in the ward area has been wet wiping with 1:250 concentration F10. Tackling contamination of the hospital environment by methicillin-resistant Staphylococcus aureus (MRSA): a comparison between conventional terminal cleaning and hydrogen peroxide vapour decontamination. J of Hospital Infection (2004): 57: 31 – 37.forbes@gwreferrals. FRENCH GL.27S. 20(8): 909 . FOSTER AP and DEBOER DJ. Unit 10. 5. Arles 2005. Swindon.co. 2002: 92:16S .18. Falco 2001. United Kingdom Email: n. BENHEIM U and DAVIDSON I. Berkshire House. SHANNON KP. County Park Estate. which is left to dry naturally. CHITTY J. 4. 4 CITATION INDEX 1. F10 Fogging Disinfection: 1:250 dilution of F10 was delivered using a commercial poultry house fogger (RL Corporation Model 1037BR Pro-ULV) for a 15 minute period. A novel disinfectant in psittacine respiratory disease. J Applied Microbiol Symp Suppl. Monterey 2002: 25 .27. The role of Pseudomonas in canine ear disease. Proc Eur College Avian Med Surg. 2. 3. et al. The results will be presented and discussed at the conference and will be published in the near future elsewhere. 3 RESULTS AND DISCUSSION The authors regret that due to technical and mechanical difficulties the results are unavoidably delayed and not available at the time of going to press. Comp Cont Ed 1998. Bacterial target sites for biocide action. SN1 2NR. MAILLARD JY. 6. VERWOERD D. Shrivenham Road. Molecular Identification and prevalence of psittacine circovirus in Israel. AUTHORS ADDRESS Great Western Referrals.918. OTTER JA. Proc Assoc Avian Vet.uk 313 . Aerosol use of a novel disinfectant as part of an integrated approach to preventing and treating aspergillosis in falcons in the UAE. 17: 17 . In this presentation the changes in the lungs and the effects of hypoxaemia on other organs. In this presentation the pathology of an outbreak of toxoplasmosis in crowned pigeons (Goura scheepmakeri sclaterii) will be used to demonstrate the acute pulmonary lesions and the consequences for other organs. 1 INTRODUCTION Severe respiratory dyspnoea is a common clinical symptom in birds.Liver pathology . RATCLIFFE and WORTH 1991). Toxoplasma gondii is a species of coccidian protozoa comprising intracellular parasites in many organs and tissues of birds and mammals. This results in decreased lung compliance. Acute pulmonary damage is often a non-specific reaction to injury of respiratory epithelial and endothelial cells from a variety of acute insults. intestines. hypoxaemia. e. Utrecht University. The only known complete hosts are cats and other Felidae (GARELL 1999.Kidney failure . The “normal” 314 . The pathophysiological cascade of a hypoxaemia will be demonstrated and based on the changes a clinical approach to counter the effects of these changes is discussed. Dorrestein. DVM. TACKAERT-HENRY and KAGERUKA 1977). spleen. Section of Pet Avian. The Netherlands CONSEQUENCE OF SEVERE DYSPNOEA FOR ORGAN FUNCTION G.Department of Pathobiology.Toxoplasmosis . including humans. Exotic Animals and Wildlife Division. and multiple organ dysfunction. kidneys and lungs (POELMA and ZWART 1972. The “normal” approach is to provide oxygen and attempting to “remove” the causative insults.Birds ABSTRACT Severe respiratory dyspnoea is a common clinical symptom in birds.g. PhD KEYWORDS Dyspnoea . This will be done by presenting details of an acute outbreak of toxoplasmosis in crowned pigeons (Goura scheepmakeri sclaterii). The diagnosis is normally made at necropsy by demonstrating the Toxoplasma-cysts and –pseudocysts by light microscopy using immunohistochemical staining. M. In acute cases lesions can be seen in many organs. The acute changes in the lungs may play a crucial role in the acute and high mortality. liver and kidneys are illustrated. including necrotic lesions in liver. but in general liver. It is a complex response to the invasion of the Toxoplasma parasite. Based on the changes a clinical approach to counter the effects of these changes is discussed. This effect will decrease the potential for reaction and effective action in these organs. In the brain no lesions were found. In liver. leading to an arterial hypoxaemia and a decreased supply of oxygen in all other tissues. 2 CASE REPORT Unexpectedly. On histological examination peracute changes were seen in the lungs including hyperaemia and vascular damage leading to transudate and haemorrhages. There is arteriolar dilatation resulting in hyperaemia. It will also support the development of local necrosis. 3 DISCUSSION 3. In time the permeability increases and the fluids change from watery transudate (oedema) to protein-rich. At post mortem all birds were in good to excellent condition. Other areas of the lung were emphysematous.approach in acute respiratory distress is providing oxygen and trying to “remove” the causative insult. The increased permeability of the microvasculature is demonstrated by fluid in the extravascular tissues resulting in oedema and fluid in the parabronchi. the other three birds were found dead in the morning without previous clinical symptoms. 4 Scheepmaker’s crowned pigeons (Goura scheepmakeri sclaterii) housed in the same enclosure in a zoo died within 4 days. 315 . In all examined organs immunohistochemically positive organisms for antibodies against Toxoplasma gondii were demonstrated. heterophils and macrophages.1 Acute changes in the lungs The acute reaction in the lungs is an excellent example to demonstrate what is going on in the body of the infected bird. increased distance between air (O2) and blood (CO2). two blue-winged kookaburras (Dacelo leachii) and two wattled brush turkeys (Aegopodius arfakianus). Through the microscope we see all the typical local manifestations of an acute inflammation. spleen and kidney were degenerated (pale) and swollen. kidney and spleen acute to subacute inflammations with necrosis were seen. marbled areas with oedema and a foamy content in the bronchi. some cytokines and trombocyte-activating factors induce this increased vascular leakage (BOCHSLER and SLAUSON 2002). In the intestinal wall and pancreas a cellular infiltration without necrosis was seen. leukotrienes. cellcontaining exudate. A peripheral margination of white blood cells can be seen in capillaries and venules. This cellular component existed of lymphocytes. The most prominent macroscopically recognisable alteration in all pigeons was seen in the lungs. complement fragments. prostaglandins. It is an overlapping series of events that form a continuum. Direct damage or a scala of mediators like vaso-kinins. 2 juvenile and their parents were housed in a mixed exhibit with two yellow-tailed black cockatoos (Calyptorhynchus funereus funereus). All acute changes lead to local stasis of the circulation. Changes of other organs were less consistent. The first reactions are haemodynamic changes. Excessive cellular responses led to severe pulmonary damage. The birds. All lungs showed dark red. One bird was found moribund. involving many systemic responses. which results in acute renal failure (JENNETTE and SPARGO 1999). Remove extravascular fluids from the lung as soon as possible to allow minimal distance between air. because of the vital function of the lung for the O2/CO2 exchanges all therapeutic measures.2 Effects on other organs In the liver the infection with the parasite resulted in an acute to subacute inflammation. The inflammatory process itself is aimed at maximising normalcy as an endpoint. This will also reduce the development of permanent scar tissue after recovery. However. the host may suffer more from tissue damage as a result of an inflammatory reaction than from damage caused by the initiating stimulus had there been no inflammatory response at all. even immunocompromised patients may benefit from treatment with chemotherapeutic drugs (such as pyrimethamine plus sulfadoxine. Tubular injury is focal and is most pronounced in the proximal tubules. The centrolobular zone. An example is seen in calves: if neutropaenia is induced by experimental means in calves with an induced pneumonic pasteurellosis. is most distal to the blood supply from the portal tract and normally has a low oxygen tension. Acute inflammatory cells accumulate in the necrotic zones. In the kidney are the tubular epithelial cells. particularly sensitive to hypoxia and anoxia. 2002). This leads to acute tubular necrosis. Microscopically.3. In many situations.and blood capillaries by stimulating kidney activity (diuretics?). control or otherwise modify inflammation (NSAID’s?). Ideally this can be accomplished by removal of the initiating stimulus (in this case: Toxoplasma organisms) and regenerative repair of the injured tissue. fibrin. inflammations can be harmful as well. and maybe also in this lung reaction. The arterial hypoxaemia at this stage cannot be reversed by simply increasing the oxygen tension of the inspired air. or trimethoprim plus sulfamethoxazole) (DIRIENZO et al. the low oxygen supply leads to ischemic necrosis of the centrolobular hepatocytes. 3. which cause rapid depletion of intranuclear ATP in the tubular epithelium. a severe but potentially reversible impairment of tubular epithelial function. 316 . the calves are partially protected against the development of severe pneumonia and the resultant hypoxia (BOCHSLER and SLAUSON 2002). coagulative necrosis of centrolobular hepatocytes is accompanied by frank haemorrhage. should be aiming at: Preventing free fluids to come into the respiratory tissue (drowning effect) by using anti-inflammatory drugs to limit. Mostly no pathological changes are seen in the glomeruli or blood vessels. in addition to providing extra oxygen. with their high rate of energy-consuming metabolic activity and numerous organelles. When the diagnosis is made in time. exudate and so forth) may be worse than tissuedamaging contributions of the original Toxoplasma infection. In the case with the pigeons the intensive inflammatory reaction in the lung with tissue damaging contributions of the inflammatory host-defence mechanisms (hyperaemia.3 Discussion of a clinical approach Although inflammation fundamentally is a defensive reaction. referred to as zone 3 in the functional concept of the hepatic acinus. However. DVM. 131 . Dorrestein. Dept. 3. The Kidney. 55: 29 .dorrestein@vet. Toxoplasmose bij kroonduiven en andere vogels in de Koninklijke Rotterdamse Diergaarde “Blijdorp”. Philadelphia: LippincottRaven 1999. TACKAERT-HENRY MC and KAGERUKA P. 18: 89 . Inflammation and repair of tissue. 7. Utrecht University.245. In: FOWLER ME.901. AIDS Res Hum Retroviruses 2002. 69: 163 .135. of Pathobiology. Goura cristata Pallas et Goura victoria Frazer. Acta Zool Path Antverpensia 1972. Toxoplasmosis of wild birds and mammals. VAN DER HORST C. du Zoo d’Ánvers. Efficacy of trimethoprim-sulfamethoxazole for the prevention of bacterial infections in a randomized prophylaxis trial of patients with advanced HIV infection. 3rd edition. BOCHSLER PN and SLAUSON DO.168. 141 . RATCLIFFE HL and WORTH CB.nl 317 . DIRIENZO AG. In: RUBIN E and FARBER JL (eds): Pathology.m. Chapter 16. FINKELSTEIN DM and FRAME P. JENNETTE J C and SPARGO BH. PhD Faculty of Veterinary Medicine. AUTHORS ADDRESS Gerry M. 5. GARELL DM. 900 . Une Épizootie de Toxoplasmose parmi les pigeons couronnés. St Louis: Mosby 2002. 2. 4. Current Therapy 4.94. POELMA FG and ZWART P.uu. 27: 655. Toxoplasmosis in Zoo Animals. Am J Path 1991. Philadelphia: WB Saunders Co 1999. MILLER RE (eds): Zoo & Wild Animal Medicine. Section Pet Avian. 3584 CL Utrecht. Acta Zool Path Antverpensia 1977. 6. the Netherlands Email: g. In: SLAUNSON DO and COOPER BJ (eds): Mechanisms of Disease.40. Yalelaan 1.4 CITATION INDEX 1. Exotic Animals and Wildlife. Diplomate ACVP KEYWORDS Birds . The reported clinical signs range from unexpected death to weight loss and passing intact seeds in the droppings. certified in avian practice. and mixed species aviaries are common recent stresses. Diplomate ABVP. psittacines. They were trimmed. Fifty three cases from 13. United States of America A REVIEW OF AVIAN ENDOVENTRICULAR MYCOSIS SUBMITTED DURING 1998-2004 D. 53 cases of endoventricular mycosis were identified. and galliformes (GRAHAM 1994). Follow-up information was obtained by mailing an inquiry form to referring veterinarians or by telephone conversation. Although antibiotic therapy is typically associated with secondary yeast infections.950 avian histology samples collected over six years were identified from the database of Zoo/Exotic Pathology Service. 1 INTRODUCTION The lesion of endoventricular mycosis is a common one of the ventricular koilin in finch and finch-like birds (HUBBARD 1985).950 psittacine biopsy samples submitted. The information requested was 318 . It is also described in ratites.Ventriculus . crowded housing. this is not a frequently reported occurrence with endoventricular mycosis (HUBBARD 1985). California. reproductive activities. Tissues were submitted fixed in 10% buffered formalin. DVM. and stained with hematoxylin-eosin.Zoo/Exotic Pathology Service Sacramento. All cases were necropsy findings and ante-mortem diagnosis and therapy in the live bird is not reported. sectioned at 5 µm. embedded in paraffin. Reavill. 2 MATERIALS AND METHODS From 13. Various conditions such as recent shipping.Yeast .Koilin ABSTRACT Endoventricular mycosis is a proliferation of yeast-like organisms within the koilin of the ventriculus. One case each of malignant lymphoma. zebra finch (Poephila castanotis). There were 12 males and 13 females when the sex was known or reported. Many birds (n = 31) had concurrent diseases. and one each Amazon (Amazona species). and proventricular dilatation disease (n = 2) were the next most common diseases. Smaller number of other species included blacksmith plover (n = 2). guinea fowl and a quail (Coturnix coturnix japonica). only 1 case reported blood work that included anemia and decreased serum protein. fungal pneumonia (n = 3). canary (Serinus canaria). Two macaws had proventricular dilatation disease. purple Grenadier (n = 2). Based on follow-up of the other avian species (42/44).35 years. age. and three birds (a cockatiel. flagellates. any ancillary tests. red-tail laughing thrush (n = 2). one cockatiel had chronic crop stasis. The nine psittacines were cockatiels (Nymphicus hollandicus) (n = 3). mockingbird (n = 2) and one each bronze-winged mannikin (Lonchura cucullatus). Other parasitic infections included protozoa (coccidia. honeycreeper and starling. orange cheeked waxbill (Estrilda melpoda) (n = 2). macaws (Ara species) (n = 3). As the disease. 3 RESULTS The most common birds with a diagnosis of endoventricular mycosis were passerines (n = 38) composed of finches (species not provided) (n = 7). and one each flamingo. cordon bleu (Uraeginthus bengalus) (n = 5). goldenfronted leafbird (Chloropsis aurifrons). and systemic mycobacterium were recognized. was not identified ante-mortem in any of the birds. and any other therapies. sex. there were 20 finches from aviary collections. cytomegalovirus. and three were wildlife rescues. red-crested cardinal (Paroaria coronata).for the species. one conure had chronic cloacal papillomatosis. Amyloid that was recognized only in the finches (n = 5). duration of clinical signs. bird of paradise. Nematode infections were most common (n = 9). Gouldians (Poephila gouldiae) (n = 7). The common clinical signs reported included significant weight loss (n = 21) and whole seeds passing in the droppings (n = 3). white-headed munia (Lonchura maja) (n = 2). The age range was from 3 days to 24 years with an average of 4. and cryptosporidia) (n = 5). Three of the finch aviaries reported using multiple water-based drug therapies. trumpeter swan. no specific therapies were attempted. megabacteria or gastric yeast (n = 4). conure (Aratinga species) and parrotlett (Forpus species). eight of the nine psittacines had other serious diseases and conditions. and macaw) were less than one week old with systemic diseases. In only four birds were gross lesions noted in the proventriculus of dilation. the Amazon had systemic mycobacterium. parrotlett. From the responses and histopathology. 319 . endoventricular mycosis. Of the 50 survey responses. survival. owl finch. 16 birds were from zoo collections. and Tippit. SCHMIDT R. Many of these birds were under significant stress. Puzio. Inflammation is usually mild. and fecal contamination. United States of America Email: Dreavill@zooexotic. Roose. 2. et al. Lane. Proc Assoc Avian Vet. Zoo/Exotic Pathology Service. hypovitaminoses (especially Vitamin A). either induced by other diseases or husbandry situations (large aviaries. stale. Fungal organisms (usually Candida sp. 279 . Fisher. 21: 25 . and gross changes are rarely seen. DVM. Riggs. whereas yeast infections in psittacines are more commonly isolated to the ingluvies. Diplomate ACVP. Ante-mortem blood work was uncommon in these birds as many were finches in large collections or zoo birds. The only gross lesion noted in our submissions was of proventricular dilation. Fronefield. Littlehale. Ristich. Citrus Heights. Histologically. Mauroo. Rich. Dazen.28. Known factors predisposing avian species to yeast infections include: prolonged antibiotic therapy.4 DISCUSSION Endoventricular mycosis is seen in a variety of pet birds and is especially common in finches. It is interesting that the majority of endoventricular mycosis cases were in finches. may bleed causing anemia or rarely a fatal blood loss. Although antibiotic therapy is typically associated with secondary yeast infections. Govers. Nye. Sitinas. feeding spoiled. or sour foods. Orosz. Only three finches came from aviaries reporting multiple water-borne drug therapies. although two of the birds had proventricular dilation disease. malnutrition. Fudge.com 320 . 7647 Wachtel Way. HUBBARD G. CA 95610. AUTHORS ADDRESS Drury Reavill. Fungal infections of ventriculi in captive birds. a stressful environment with moist floors.) are found in the koilin layer and occasionally in the mucosa.282. Okimoto. J Wildl Dis 1985. Endoventricular mycosis: an avian pathologist’s perspective. GRAHAM D. Swerida. It is this fragmentation of the koilin and possible gastric dysfunction that may have resulted in the proventricular dilation. where individual testing was not performed. Only one bird had ante-mortem blood work and it was anemic. Diplomate ABVP. or wildlife rehabilitation). dirty nests. 5 CITATION INDEX 1. Fiskett. fragmentation of the koilin was a common associated lesion of yeast infections of the ventriculus. These lesions. Reno 1994. Douglass. EISENBRANDT D. like those of bacterial infections of the koilin. 6 ACKNOWLEDGMENTS Drs. Stonebreaker. many of our cases received no specific therapy. and co-existing bacterial or viral infections. zoological displays. Burke. MacCabe-Laduke. certified in avian practice. 13 Ara ararauna (12 chicks and an adult). E. in juvenile birds (3 to 12 weeks of age). in some cases. The mortality peak in macaws occurs from 4 to 8 weeks of age. Avian polyomaviruses (APV) that infect budgerigars. Tarantino3. R. and 11 Ara spp. Bert4. No concurrent PBFD infection was detected in analysed macaws. 3 Ara spp. In the literature descriptions of juvenile and adult macaws APV infections are infrequent compared to budgerigar and lovebird cases. Private practitioners2. no reports of macaw deaths related to APV infection are described in Italy. and each report involves one or few subjects.University of Camerino. transmission electron microscopy and immunogold techniques. University of Pisa. C. after which infection is asymptomatic.Department of Animal Science1. (eggs containing embryos of different age). in all cases. viral presence was revealed by immunohistochemistry.5 to 3 years of age). other psittacine birds and finches appear to be morphologically and. Department of Animal Production4. Rossi1 KEYWORDS Avian polyoma virus .Macaws . G. characterized by high mortality in embryos. Pesaro1. and in some adults (1.and microscopic lesions were typical of APV infection and. Italy THE FIRST ITALIAN OUTBREAK OF POLYOMAVIRUS INFECTION IN MACAWS S. In most cases PCR confirmed these tests and was the only test performed in eggs/embryos. until now. In addition.Outbreak . Ceccherelli2. In total 10 Ara chloroptera (9 chicks and an adult). 1 INTRODUCTION Polyomaviruses and papillomaviruses are members of subfamilies of the Papovaviridae. Scoccianti2. S. Department of Animal Pathology3. University of Turin.Immuno-electron microscopy Polymerase chain reaction ABSTRACT Avian polyomavirus (APV) is one of the most significant viral pathogens of cage birds. antigenically 321 .(chicks of 2 weeks of age). Macaws are susceptible to APV infection and disease is seen up to approximately 14 weeks of age. were analysed. We present the description of an outbreak of APV infection in three different breeding farms of the central area of Italy. Epidemiology and Ecology. Macro. were necropsied to identify the aetiology of the outbreak. anorexia. 1991). Samples of liver. in a period of six months. 1991). heart. 1991). Sections were also mounted on positively charged glass slides (Superfrost Plus. 1986). In addition. and in some adults (1. and embedded in paraffin. and onto Campylobacter Kit Blaser (Difco Laboratories). the clinical presentation. intestine. distribution of lesions and epizootiology of these viruses differ dramatically among susceptible species (CLUBB et al. Italy). These animals belonging to three different breeding farms of large psittacine birds of the central area of Italy. until now. the faecal material was inoculated onto Desoxycholate Agar (Difco Laboratories.5 to 3 years of age) was observed.similar. Pittsburgh. For the latter purpose. fixed in 10% buffered formalin. no reports of outbreak of macaws death related to APV infection are described in Italy. evidence suggests that the genome of the virus that infects various psittacine birds and finches partially differs.subclinical infections in the natural immunocompetent host.e. Detroit. 10 (Ara chloroptera) (9 chicks and an adult).) (eggs containing embryos of different age) were analysed in our laboratory (Dept. 13 (Ara ararauna) (12 chicks and an adult). central nervous system and skin were collected from all birds. lungs. routinely deparaffinized and then immersed in distilled water containing 15% H2O2 for 30 min at room temperature in order to inactivate endogenous peroxidases. Fresh smears of intestinal content were made and examined both for parasites and bacteriological tests. bursa. 3 (Ara spp. The slides were 322 . Michigan. the acute nature of some avian polyomavirus infections is typical of bird infection. However. pancreas. Because of the differences in the way avian polyomavirus affects budgerigars versus larger psittacine birds. With regards to polyomavirus infections in mammals. Antibody surveys indicate that most infections in non-budgerigars psittacine birds are sub-clinical. and the immune response that occurs in these birds prevents the acute form of the disease. generalized infection associated with APV was described in young budgerigars and was called budgerigar fledgling disease (BFD) (LEHN et al. RITCHIE et al. 2 MATERIALS AND METHODS During 2003. The most important information about APV infection in macaws derived from experimental infections and only sporadic descriptions of natural APV infection. delayed crop emptying. 3 µm sections were stained using haematoxylin and eosin and periodic acid Schiff’s reaction (PEARSE 1985) for histological examination. Blood smears were stained with May Grünwald . Fisher. characterized by apathogenic . USA).Giemsa for haemoprotozoan examination. the extensive information derived from studies in budgerigars may not be completely applicable to other psittacine birds (RITCHIE et al. All birds. Additionally. spleen. 1984. bleeding under the skin. Camerino. The first acute. characterized by per-acute death with no premonitory signs or following a 12-48-hour period of clinical changes (i. diarrhoea. polyuria and dyspnoea) are reported (RITCHIE et al. depression. PA). kidney. in which an outbreak characterised by high mortality in embryos. and 11 (Ara spp. recently dead. some stool samples were enriched using Selenite Broth (Difco Laboratories) and then inoculated onto SSA (Salmonella-Shigella Agar) and BGA (Brilliant Green Agar) (Difco Laboratories). in juvenile birds (3 to 12 weeks of age). of Veterinary Sciences.) (chicks of 2 weeks of age). Warrington. and floated on drops of 10% normal goat serum. postfixed in phosphate buffered 1% OsO4 and. followed by a proteinase K (DAKO Corporation. Finally. followed by incubation with 3. Amplification for BFDV: the amplification was carried out with primers n°2 and 4° described by YPELAAR et al. PCR products were separated by 323 . In order to minimize non specific background staining. Primer antisense (n°4): GTC ACA GTC CTC CTT GTA CC. 1.2% saponin for 60 min. The final reaction conditions were as follows: 50mM KCl.rinsed in phosphate-buffered saline (PBS) three times for 2 min each. UK). UK) for 45 min at room temperature. An initial denaturing step at 94°C for 2 min was followed by 32 cycles of 60°C for 30s.1% bovine serum albumin (BSA) plus 0.5mM MgCl2. Semi-thin sections were stained with methylene blue and Azure II while for immuno-electron microscopy. For this purpose DNA extraction: DNA was extracted from animal tissues using the Wizard genomic DNA purification kit� (Promega – USA) with some modifications. Control procedures included the use of a known polyomavirus infected tissue. MA)..05% Tween 20 in PBS (BTP). or non immune mouse ascitic fluid (both diluted 1:10 in BTP containing 1% normal goat serum and 0. Burlingame. 72°C for 1 min and 94°C for 1 min. Negative controls included normal bird tissues and positive controls included psittacine bird tissues infected by APV. 10 mM Tris-HCl pH9... Primer sense (n°2): AAC CCT ACA GAC GGC GAG. Carpinteria. 100 ng of each primer and 0. Tissues and embryos at different stage of development were also sampled and utilised for PCR examination.3’diaminobenzidine solution containing H2O2 as substrate-chromogen for 5 min. The sections were then incubated with primary anti-VP-1 antibody (KHAN et al. Peabody. CA) treatment at room temperature for 6 min. Chemical Co.2% saponin. rinsed in PBS and incubated with a biotinylated anti-mouse secondary antibody for 45 min at room temperature. Pa). the sections were incubated in a serum blocking solution (10% non-immune horse serum in 1% bovine serum albumin dissolved in PBS) for 1 hr at 37 °C. 200mM of each dNTP. The slides were rinsed in distilled water. spleen and kidney were immediately fixed in phosphate buffered 0. Missouri) diluted 1:50 in BTP containing 1% normal goat serum and 0. embedded in Epon/Araldite (Polyscience Inc. and 0. (1999).15 units of Taq polymerase (Promega – USA). 0. A final run of 72°C for 7 min completed the program. St. Sections were then incubated for 24 hr at 4°C with either anti-VP-1 mouse monoclonal antibody (6). sections were incubated with enzyme conjugate (ABC-peroxydase complex. Vector. that amplified a 717 bp tract of the BFDV ORF1. Stansted. portions of liver. 2000) diluted 1:500 in PBS overnight in a 4 °C moist chamber.2% saponin). ultra thin sections (70nm) were placed on 200-mesh nickel grids supplied with formvar-carbon film (Agar Scientific Ltd. In addition. etched with sodium metaperiodate for 30 min. rapidly counterstained with haematoxylin and examined microscopically. The PCR amplification was carried out in a total volume of 25 µl. the sections were incubated with goat anti-mouse immunoglobulin G gold conjugated. Grids were then fixed in 1% glutaraldehyde and stained with uranyl acetate and lead citrate and examined with a JEOL 1200-EX transmission electron microscope (JEOL. Louis.4. 10nm (Sigma. After three rinses in PBS. after dehydration.1M 2% glutaraldehyde pH 7. PCR was performed in a GeneAmp PCR System 2400� (Applied Biosystem -USA) thermal cycler. Amazon parrots (Amazona spp. In some cases inflammatory cell infiltrates. Histological examination evidenced in all birds various tissue lesions characterized by the presence of typical karyomegalic. Especially in the liver. lovebirds (Agapornis spp. especially budgerigars (Melopsittacus undulatus). but in these cells intra-nuclear inclusions were less frequently observed. In addition. (1991). 3 RESULTS Similar lesions were observed in all necropsied birds. immuno-histochemistry documented APV infection in damaged tissues belonging to all dead birds. in some cases. the presence of intra nuclear inclusions was accompanied by multi-focal hepatocellular necrosis and haemorrhages. In the spleen as well as in the bursa.).and hepatomegaly with irregular red and yellow mottling of the liver. The positive reaction was characterized by intense brown chromogen deposition in large inclusions. according to the procedure suggested by the authors. heart. all birds had abdominal distension with spleno. in the same tissues inclusion bodies were present within intimate cells of arterioles. amphophilic intra nuclear inclusions. Amplification for APV: The amplification was performed using primers which have been published by PHALEN et al. Similar results were obtained in immuno-electron-micropic tests. consisting in multi focal heterophilic to lympho-plasmacitic cells. strictly associated with hyper-plastic aspect of the Kupffer cells. accompanied by interstitial inflammatory infiltrates. No bacterial organisms were isolated from cultures prepared with material collected on necropsy.) and African 324 . suggestive of an APV infection. A large number of birds showed severe thickening of the intestinal walls with haemorrhage.to 49 nm viral particles inside intra-nuclear inclusions. in skin and skeletal muscles. which demonstrated typical endo-nuclear inclusions. Using monoclonal VP-1 specific antibody. Similar aspects of variable degrees of epithelial necrosis. surrounded the areas of hepato-cellular necrosis. severe aspects of depletion and lymphocytic apoptosis phenomena were observed. indicating the antibody recognition of VP-1 specific of APV. cockatiel (Nymphicus hollandicus). in which endo-nuclear aggregates of viruses were labelled by gold particles. In the most of affected birds. 4 DISCUSSION Acute and chronic epornitics of avian polyomavirus (APV) infection have been observed in various genera and species of juvenile and adult psittacine birds. kidneys were pale and swollen and hydropericardium was also evident. meninges and.electrophoresis for 45-60 min at 7-10V/cm in a 2% agarose gel stained with ethidium bromide. The lungs were generally congested and contained an exudate. PCR results demonstrated the presence of DNA of polyomavirus in different organs of chicks and in embryos. All negative control tissues were devoid of chromogen deposition. Ultra-structural examination of affected tissues demonstrated the presence of aggregates of a great number of 42. were also observed in renal tubular epithelial cells. Haemorrhages were also observed in spleen. various degrees of myocardial necrosis were detected in of necropsied birds. JHONE R. Polyomavirus persistent infections are frequently described in mammals and in birds. IHC-positive inclusion bodies. painted conures (Pyrrhura picta). when various causes provoke a similar deficiency in the immune system. Boulder 1984: 121 – 129. this is the first report of an acute APV outbreak in embryos and young macaws in Italy. and the peak mortality occurs from 4 to 8 weeks of age. moluccan cockatoos (Cacatua moluccensis). depletion and death of lymphocytes in the bursa. the most important observations are performed during experimental infections and very little is known about the possibility of APV embryos infection. no reports regarding APV outbreak in macaws are described in Italy. Proc Assoc. graycheeked parakeets (Brotogeris pyrrhopterus). we can conclude that polyomavirus-induced lesions suggested an immune-mediated disease. blue and gold macaws (Ara ararauna). 1995). 2. In previous reports. eclectus parrots (Eclectus roratus) and pionus parrots (Pionus spp. 1993. quaker parakeets (Myiopsitta monachus). as reported by different authors (PHALEN et al. In addition. J Virol Methods 2000.gray parrots (Psitacus erithacus). 1993). BECK I. 1992. 89. In addition. different Authors describe clinical signs. and confirms that young macaws were very susceptible to avian polyomavirus acute and symptomatic infection. when stress or concomitant diseases intervene. 1995). shed virus resulting in outbreaks of disease (PHALEN et al. According to the literature (PHALEN et al. 1992. On the basis of the macro and microscopic findings. rose-ringed parakeets (Psittacula krameri). Therefore. Avian Vet. 1993). necrosis of skeletal and myocardial muscular cells. et al. 1995) the most relevant lesions observed in necropsied birds were represented by vasculitis (related to diffuse haemorrhages) with presence of inclusion bodies in the walls of blood vessels. RITCHIE et al. generalized vasculitis. and immune-mediated hepatitis.) (RITCHIE et al. 1991). Several studies suggest that these subclinically infected birds. and the histological findings are related to high prevalence of immune-complex production during the disease. In macaws. and histological lesions observed in one or few animals during spontaneous episodes of APV infection. Our report. membranous glomerulopathy with focal necrosis (and rare inclusion bodies). 39 – 48. necroscopic findings. KHAN MS. RITCHIE et al. CLUBB SL and DAVIS RB. confirms the susceptibility of macaws to APV infection and the fact that the disease up to approximately 14 weeks of age. Development of a blocking enzymelinked immunosorbent assay for the detection of avian polyomavirus-specific antibodies. particularly in budgerigars (PHALEN et al. necrotic foci in spleen parenchima and hepatic foci of centrilobular necrosis with abundant. It is possible to explicate the tissue lesions represented by membranous glomerulonephritis. this might be responsible for the occasional APV-induced death in adult persistently infected psittacine birds or the frequent deaths that occur in some born infected young birds (RITCHIE et al. white bellied caiques (Pionites leucogaster). Outbreak of papova-like viral infection in a psittacine nursery – retrospective view. 5 CITATION INDEX 1. splendid parakeets (Neophema splendida). 325 . L. 151. NIAGRO FD. LATIMER KS and DAVIS RB.it 326 .1037. 8. J. Avian polyomavirus: an overview. WILSON VG and GRAHAM DL. PHALEN DN. 735 . 362 . Cloning and characterization of budgerigar fledgling disease virus (BFDV). Via della circonvallazione 93/95 62014 Matelica MC. an avian Polyomavirus.rossi@unicam. Avian viruses: function and control. Italy Email: giacomo.K. LEHN H and MULLER H. University of Camerino. HUDELSON S. DVM.832.. A universal polymerase chain reaction for the detection of Psittacine Beak and Feather Disease Virus. GE WILCOX and RAIDAl SR. 6. PEARSE AGE. 10. 7. Avian polyomavirus infection and disease: a complex phenomenon. 1995. RITCHIE BW and CARTER K. MATTEWS J. RITCHIE BW. Proc Eur Assoc Avian Vet.10. Histochemistry. Vet Microbiol 1999.167.370. 5: 147 . WILSON VG and GRAHAM D. Rossi. J Clinical Microbiol 1991. PHALEN DN. Lake Worth: Wingers Publishing Inc. 1985. WILSON VGH and GRAHAM DL. Avian polyomavirus biology and its clinical applications. and HUDELSON P. 4th edition. AUTHORS ADDRESS G. YPELAAR I. Edinburgh: Churchill Livingstone Inc. 1991.148. Analytical technology. PHALEN DN.153. Proc Assoc.216.. Utrecht 1993: 200 . Virology 1986. Avian Vet. (eds). Polymerase chain reaction assay for Avian Polyomavirus. Assoc. Avian Vet. MR BASSAMI. theoretical applied. 4. 136 .3. 5. 1030 . PhD Department of Animal Science. 9. 68: 141 . New Orleans 1992: 5 . 1 INTRODUCTION The cloaca is comprised of three areas: the coprodeum. The coprodeum is the connection between the distal colon and the cloaca. Dipl ABVP (Avian) KEYWORDS Cloaca – Avian – Coprodeum – Urodeum – Proctodeum – Cloacoscopy Cloacotomy ABSTRACT The cloaca is a complex structure in the bird. K. Boston. Specific case examples are included for each of these categories. Anseriformes. prolapse. The coprourodeal fold prevents faecal contamination of the urodeum and proctodeum and during defecation protrudes through the vent to serve this function. obstruction and neoplasia. It functions as a holding and processing area for products of the gastrointestinal. A fold within the cloacal lumen called the uroproctodeal fold separates the urodeum and the proctodeum. The anatomy and physiology of the cloaca will be discussed as well as the diagnosis and treatment of different cloacal conditions. DVM. The bursa reaches its largest size at eight to twelve weeks of age and decreases in size as the bird ages.Angell Animal Medical Center. Cloacal disorders discussed will include cloacal infection and inflammation. The Bursa of Fabricius is a diverticulum of the dorsal wall of the urodeum. Ritzman. Coracopsis vasa) have phallic bodies housed within the cloaca. a copulatory organ may be housed within the cloaca. the urodeum and the proctodeum. The bursa is involved with the immune function of the bird and is the location of B-lymphocyte differentiation. The urodeum contains the openings of the ureters and the genital ducts. reproductive and urinary systems. ratites and an occasional psittacine species (Vasa parrot. Due to the complexity of the cloaca and the pertinent anatomical structures associated with it. In male birds of some species. United States of America CLOACAL DISEASE AND DISORDERS IN THE AVIAN PATIENT T. The opening and closing of the vent is controlled by striated sphincter muscle. The proctodeum communicates with the outside of the bird’s body through the vent. the health of this structure is essential to the well being of the avian patient. During the egg laying process the coprourodeal fold will protrude 327 . The author diagnosed a case of complete cloacal obstruction in an umbrella cockatoo (Cacatua alba) from a fungal granuloma. respiratory signs may be apparent due to impingement of the abdominal airsacs. tone or sensation may be present. decrease or change dropping appearance and production. Causes of cloacal abnormalities include bacterial or fungal infection. causing possible renal failure and haematological changes such as an elevation in uric acid values. inability to breed or produce eggs normally. or cloacal orifice. or mass effect in the caudal abdominal region. change in perching posture. A thorough anamnesis should include information on age. The vent. or neoplasia. PHALEN 1997). After the vent has been evaluated. With cloacal inflammation or infections a change in the protein electrophoresis pattern may occur. Masses can develop from the cloacal mucosa or associated structures. Diagnosis of Cloacal Disease Diagnosis of cloacal disease is often possible during the initial examination of the bird. Stress-induced polyuria can occur if the bird releases the cloacal contents too quickly for normal urine processing and water reabsorption to occur. Common causes of cloacal disease The causes of cloacal disease can be related to any of the systems associated with this structure. Vent stricture and subsequent cloacal obstruction can occur in avian patients secondary to distal cloacal and external vent sphincter abnormalities. faecalith or urolith formation. A cursory examination can be performed in the restrained 328 . gender. can develop into large concretions of several centimetres in size causing severe dilatation and obstruction of the cloaca. Cloacal neoplasia is an infrequent diagnosis but should be included as a differential for cloacal wall thickening or luminal masses (ANTINOFF 1997). retained eggs. lethargy. Cloacal papillomatosis can produce proliferative lesions of the cloacal mucosa (SUNDBERG 1986. Cloacal lymphoma was diagnosed by the author in an Indian ring-necked parakeet (Psittacula krameri) from cloacal mucosa biopsies obtained via cloacoscopy. prolapse of cloacal mucosa or other cloacal structures. Urine deposited from the ureters into the urodeum often moves retrograde into the rectum where this reabsorption takes place (SKADHAUGE 1968). inflammation. diarrhoea. Change in vent size or symmetry. reproductive status and activity.in a similar manner. If the cloaca becomes obstructed congestion of the ureters can occur. an internal cloacal exam can be performed. Some birds with cloacal infection may develop a heterophilic leukocytosis. A thorough cloacal examination should be performed on every bird as part of the routine physical assessment. flatulence. haematochezia. prolapse. soiling of vent area. Clinical signs of cloacal disease can vary but include tenesmus. Cloacaliths. Routine haematology including a complete blood count and chemistry profile will often be normal. should be examined prior to performing an internal cloacal exam. Most birds reabsorb water from the urine held in the urodeum. The avian patient with a blockage of the cloaca will present with clinical signs of obstructive disease (RITZMAN 1999). If cloacal disease is due to a mass effect within the cloacal lumen. diet and management. anorexia. with multiple lamellar layers. 15 minute. Any masses or changes in mucosal consistency should be further evaluated. survey radiographs can be taken. Radiography is a useful diagnostic modality in the diagnosis of obstructive cloacal disease. urate crystals. Cloacal mucosal biopsies may be taken at the time of the endoscopic examination. Timed radiographs are then taken after contrast administration.bird. appropriately sized swab should be gently inserted into the cloaca. Cloacal cytology can be useful in diagnosing disorders of the lower intestinal tract. The number and timing of the radiographs 329 . it may not be possible to safely insert anything into the cloaca. Normal cloacal cytology often includes epithelial cells (noncornified squamous or columnar). and full-thickness biopsies are contraindicated in most cases. Michael Taylor (TAYLOR and MURRAY 2002). The size of the patient dictates equipment potential. A definitive diagnosis of papillomatosis is made from histopathologic examination of a biopsy (SUNDBERG 1986.7 mm rod lens endoscope with an instrumented sheath system works well for an internal cloacal examination (TAYLOR and MURRAY 2002). lubricated. contrast radiography can then be utilised to further delineate structures. PHALEN 1997). extracellular bacteria. A whole body ventrodorsal and lateral view should be taken to allow for evaluation of the entire gastrointestinal tract. For most cases. In small birds. plant and faecal material and other background debris. The internal visual cloacal exam can be performed in several ways. Intraluminal masses and mucosal inflammation or ulceration can be detected with this swab technique. Papillomatosis of the cloaca can cause proliferative mucosal changes (SUNDBERG 1986. reproductive tract. urinary tract or cloaca itself. If survey radiographs indicate any abnormalities of the gastrointestinal system. A Gram’s stain should be performed on a faecal sample collected from the cloaca. however. 30 minute and 60 minute views are taken. An otoscope with a sterile cone tip can be used but has limited range of view. A 2. Infusion cloacoscopy is an ideal diagnostic tool for internal cloacal visualisation and described by Dr. there are several options. Internal cloacal structures such as the ureteral openings. Direct visualisation of the cloacal lumen is the most effective way to diagnosis cloacal disease. oviductal orifice and distal colon can often be examined. Swab samples for bacterial or fungal cultures can be collected. The mucosa should appear moist and pink with a uniform appearance. Care must be taken to choose a biopsy site devoid of pertinent anatomic structures. The distal cloacal mucosa can then be gently everted for examination. Application of dilute acetic acid (vinegar solution) to the cloacal mucosa has been reported to increase suspicion of papillomatous changes by causing a white discoloration of the affected mucosal tissue. Insufflating the cloaca with an infusion of a liquid such as sterile saline will aid in visualisation (TAYLOR and MURRAY 2002). In medium to large-sized birds. Cell samples collected from the cloaca may originate from any of these organ systems and additional diagnostics are required to localize abnormalities. PHALEN 1997). reproductive organs and urinary systems. Obstructive conditions in the cloaca can cause subsequent dilatation of the distal intestinal tract or changes in other related organ systems. For initial assessment. A sterile. A contrast agent such as dilute barium suspension can be administered by tube into the proximal gastrointestinal tract (crop). Retroperistaltic movement of the contrast agent into the colon is normal and will often be noted with this study. an L-shaped incision can be made. although useful. Cloacoscopy can be utilised for the biopsy of internal cloacal masses and the treatment of papillomatosis and debridement of cloacal lumen masses. into the cloacal lumen. A surgical technique has been described for the treatment of cloacal papilloma via a cloacotomy and cloacal mucosal stripping (ANTINOFF 2000). Treatment Options Cloacal conditions can either be managed medically. For this study. Care should be taken to avoid normal anatomic structures of the cloaca such as the ureteral openings or reproductive structures. there is a subcutaneous layer of fat that must be reflected prior to encountering the cloacal serosa. a fullthickness incision is made through the cloacal wall. A technique is being developed utilizing diode laser application via cloacoscopy with fluid insufflation by Dr.must be adjusted according to the patient size and condition. Stephen HernandezDivers at the University of Georgia. a cloacotomy may be required (DVORAK et al. A cloacotomy allows for direct visualisation of the cloacal lumen and facilitates removal of masses (e. Laser ablation therapy via cloacoscopy was utilised at the University of Georgia. A localized contrast study can be performed on the cloaca itself. surgically or with a combination. Gastrointestinal emptying times vary according to species and patient size. The shape of the incision made depends on the size of the patient and the exposure required. cloacaliths or eggs) or resection or biopsy collection of abnormal tissues. if possible. College of Veterinary Medicine. The optimal treatment program is dependent on the underlying cause of the condition. Fluoroscopy can be utilised to evaluate the gastrointestinal motility and function of the bird. After the contrast agent has been infused an immediate radiograph can be taken. This diagnostic test. Once the cloacal serosa is exposed. After the skin is incised. a complete visual examination can be 330 . is rarely necessary for the diagnosis of cloacal obstruction since direct internal visualisation is often diagnostic. The caudal abdomen is prepared and an incision is made over the cloaca. to prevent potential aspiration of the contrast agent. For the cloacotomy procedure. Once the cloacal lumen is exposed. and the contrast agent (usually 3.6 ml) can be directly infused. Any masses within the cloacal lumen will be outlined with the contrast agent. The thin abdominal muscle wall and pleuro-peritoneum must be incised. the patient is anesthetised and placed in dorsal recumbency. Endotracheal intubation should be performed. and references should be consulted when evaluating the contrast films.g. a flexible catheter can be inserted into the cloaca. surgical intervention if often required. College of Veterinary Medicine to treat a case of cloacal adenocarcinoma (HERNANDEZ-DIVERS 2004). Because many cloacal disorders are caused by a prolapse or mass effect within the cloaca. 1998). For maximum visualisation of the cloacal lumen and removal or resection of large masses. and in most birds. There is potential in the future for development of a bipolar cautery through the rigid endoscope that may replace the diode laser technique and be more readily available to the veterinary clinician. For maximum exposure. the subcutaneous tissues are gently dissected. Inflammation and infection of the cloacal mucosa can lead to ascending infections of the gastrointestinal tract. 3 CITATION INDEX 1. 2.100. medical or surgical prevention of egg laying may be indicated. and absorbable sponges (Gelfoam®. J Avian Med Surg 1997. HOEFER HL. ventplasty surgery can be performed in order to reduce vent size and prevent future prolapse. If indicated. Closure of the cloacal wall is made with an interrupted or continuous inverting pattern with absorbable suture. and those diagnosed with cloacal disease may require more frequent evaluations. Gentle tissue handling techniques will minimize mucosal bleeding. ANTINOFF N. If there is a history of cloacal obstruction due to egg binding in a female bird not intended for breeding. et al. egg binding and other obstructive cloacal conditions can lead to vent stretching and atony. Care should be taken to maintain adequate vent diameter to allow for normal defecation and reproductive activity. If the bird normally eats a seed-based diet. haemorrhage of the tissue is quite common. 2 CONCLUSION Cloacal disease is a common clinical entity in the avian patient. ANTINOFF N. antimicrobial and/or antifungal medications should be given if indicated. Proc Assoc Avian Vet. Because the cloacal mucosa is often inflamed or infected. Close monitoring of the droppings and the external vent area as well as regular examinations will aid in the prevention and early diagnosis of cloacal disease. or ureters. Clients can be informed on how to monitor their birds for cloacal dysfunction. Kalamazoo. Smooth muscle neoplasia of suspected oviductal origin in the cloaca of a blue-fronted Amazon parrot (Amazona aestiva). Nutritional support should be initiated as soon as the patient can be fed. Upjohn. which will allow for early diagnosis of any problems and prevention of obstruction. 11(4): 255 . Cloacal prolapse. Treatment of a cloacal papilloma by mucosal stripping in an Amazon parrot. Regular cloacal examinations are recommended for all avian patients. Portland 2000: 97 . The patient should be supported post-operatively with analgesia. If cloacal prolapse has occurred. The condition of the vent should also be assessed at the time of surgery. A readily digestible diet such as an extruded commercial avian food is recommended as the base of the diet. reproductive tract. resection of affected tissue can be performed. MI) can be placed within the cloacal lumen to control haemorrhage and promote haemostasis.259. a gradual conversion to the formulated diet is recommended. fluids and general supportive care. or in the case of papillomatosis.performed and any masses gently lifted out of the cloaca or resected. 331 . The body wall and skin are closed with a simple continuous or interrupted pattern. cloacapexy can be performed at the time of the cloacotomy. ROSENTHAL KL. Cloacal assessment is recommended at regular three to six month intervals for the first year post-cloacotomy and at least annually for subsequent years. Biopsy samples can be collected. O’BRANION MK. In: RITCHIE B. Proc Assoc Avian Vet. Comp Biochem Physiol 1968. 6. 5. RITZMAN TK. Personal communication. AUTHORS ADDRESS Tracey K. 9. Boston. The psittacine cloaca: a clinical review. BENNETT RA and CRANOR K.org 332 . Avian medicine and surgery.3. FL: Wingers. 4. Cloacal papillomas in psittacines. and MURRAY MJ. 1994. 24: 7 . and HARRISON L (eds): Avian medicine: principles and application. HARRISON G. MA 02130.312. SUNDBERG JP. PHALEN DN. Philadelphia: WB Saunders. J Avian Med Surg 1998. 1.15.932. 350 South Huntington Ave.. Cloacal storage of urine in the rooster. CLUBB SL. 310 .512. Dipl ABVP-Avian Practice Angell Animal Medical Center-Boston.18. New Orleans 1999: 87 . College of Veterinary Medicine. SKADHAUGE E. Am J Vet Res 1986. 12 (1): 11 . In: ALTMAN RB. LUMEIJ JT. 1997. Proc Assoc Avian Vet. DORRESTEIN GM. Lake Worth. Ritzman. Department of Small Animal Medicine & Surgery. 2004. DVM. 8. Obstructive cloacal conditions in the avian patient.91. et al. Viruses. 7. Monterey. University of Georgia. 509 . and QUESENBERRY KE (eds). HERNANDEZ-DIVERS S. 4.269. United States Email: tritzman@angell. JUNGE RE. Cloacotomy for excision of cloacal papillomas in a Catalina macaw. DVORAK LR. TAYLOR M. 2002: 265 . 47(4): 928 . Gastroenterology. is composed of an external part. a complex structure with the membranous labyrinth. The short canal (external acoustic meatus) leading to the tympanic membrane (eardrum) is covered by special feathers that lack barbules and do not obstruct sound transmission. In birds. This is caused by the processes of the extracolumellar cartilage. DVM KEYWORDS Birds .Ear . a complex structure with the membranous labyrinth. They are called ear coverts or auriculars. a middle part with an air filled tympanic cavity and an inner part. 333 . often hidden by specialized feathers.Clinique Vétérinaire de l’Arche. The tympanic membrane. France AURICULAR DISEASES IN BIRDS F. at the end of the external acoustic meatus. often hidden by specialised feathers. a middle part with an air filled tympanic cavity and an inner part. Valence. is composed of an external part. Rival. 1 INTRODUCTION The ability to listen and vocal communication play an important role in the natural life of birds. The avian ear. an organ of hearing and balance. an organ of hearing and balance. but the practitioner must be familiar with and be able to recognise them when encountered.Otitis . This can be observed during endoscopy examination of the canal.Tympanic membrane ABSTRACT This paper intends to describe the anatomy and physiology of the ear and the most common diseases as seen in private practice. have no pinna. protudes cone-like into the meatus. 2 ANATOMY AND PHYSIOLOGY OF EARS The avian ear. Diseases of the ear are not very common in birds. unlike many mammals. Birds possess a highly evolved auditory system and their hearing performance is not necessarily inferior to those of most mammalian species. the outer or external ear. It consists of the cochlear organ and a vestibular organ. Inner ear The inner ear is the sensory receptor for both sound and equilibrium. they are not replaced. a canary’s song would have to be slowed down ten times before the 334 . birds appear less sensitive to higher and lower vocal tones as compared to humans. though the sensitivity to loudness is reduced. but some birds are able to detect much higher frequencies (up to 10 .2. and stapes). Twenty eight days after an exposure to a chemical (kanamycin) that destroys hair cells the number of hair cells in the regenerated bird ear had nearly returned to normal levels (DOOLING and DENT 2001). the cochlear (round) window (accessory tympanic membrane) and a rod-like bone (ossicle) known as the columella. In mammals.1. Hair cells Loss or damage of sensory or hair cells in the inner ear results in hearing impairment. The infraorbital sinus in most birds opens dorsally into the middle and caudal nasal conchae and is connected by small canals in the tympanic cavity with the cervicocephalic air sacs (KRAUTWALD 1998). The columella picks up sound vibrations from the eardrum and transmits them by the footplate at the proximal end to a membranous oval window in the inner ear. Their ability to differentiate various sounds is ten times faster than humans.2.1. ligaments. Hearing ranges of birds The structure of a bird’s ear is simpler than in humans. In other words. hearing is best at frequencies of 1 . but their hearing seems to be about the same. 3. Middle ear The middle ear is an air filled tympanic cavity containing a muscle. have the remarkable ability to regenerate and replace the hair cells of the inner ear following damage or loss (DOOLING and DENT 2001). so hearing loss is permanent. The cochlea is not coiled as in mammals but slightly curved.2. incus. • • For most birds. Birds differ from mammals in that they have only one bony ossicle (the columella is equivalent to the stapes) while mammals have three (malleus. However. The cochlear organ contains the basilar papilla as a sensory epithelium for hearing. Birds. however. This single ossicle consists of two segments: a cartilaginous extracolumella and the bony columella proper. The ability of birds to discriminate differences in the frequency of sounds and to detect gaps between sounds is generally similar to humans. The range of frequencies (pitch) is similar. once these hair cells are lost.12 kHz). 3 SOME AVIAN CHARACTERISTICS 3. These cells can be damaged by overexposure to intense sounds. the tympanic membrane.4 kHz. 2. Most birds. where the brain compares information received from each ear and translates subtle differences into a perception of a single sound coming from a particular location.human ear could identify all the notes that are learned by a canary chick. including parrots. one opening higher than the other. Some birds can echolocate by producing specific sounds. • • There is considerable evidence that birds can and do localise the vocalisations (or other sounds) of their own and other species. prey). Sound localization Birds have good hearing but they hear things differently to humans. Echolocation • Echolocation occurs in 2 families of birds: Steatornidae (oilbirds) and Apodidae (cave swiftlets). the ear canals are covered by a thin membrane that has tiny openings evident by day 2.3. 335 . but owls excel at this task. In many macaws and eclectus parrots the ear membrane opens very slowly creating pinpoint holes that dilate fully in four to ten days.5. This also means that they do not hear their own wing beats. 3. Ear opening The outer ear canal membranes are open at hatching in cockatoos and king parrots. pinpointing where sounds are coming from requires a complex process called binaural fusion. These birds exhibit such extraordinary sound localization abilities that they are able to hunt in total darkness. which means sounds appear louder in one ear than the other and helps to determine the distance and direction of the source of the sound. Barn owls (Tyto alba) can locate prey by sound alone and this remarkable sensitivity and accuracy is achieved by having: • • • A facial disc (like a satellite dish) that funnels sound to the eardrums. For birds it is important to localize the vocalisations or other sounds (and therefore a prey). 3. The ability to identify where sounds are coming from based on auditory cues is common to all hearing creatures. 3. but do not begin to open until approximately 23 days. Many owls are extremely capable of localising the source of sounds (and therefore. i. Ear openings that are asymmetrical. They can locate a mouse by its sound. • Penguins also appear to hunt by echolocation while underwater. • Birds (and other animals) echolocate by producing clicking sounds and then receiving and interpreting the resulting echo. Flight feathers that have soft rounded edges allowing the owl to fly silently. The ear canals of macaws are also covered at hatching.e. In eclectus parrot (Eclectus roratus) hatchings.4. do not hear ultrasonic vibrations. While most sounds can be distinguished quite well with one ear alone. • Used for orientation in dark caves. 1. 4.5. Benign neoplasia of unspecified type was reported to be the cause of haemorrhage in one Amazon parrot. Pruritus may be present causing the bird to scratch the ear or rub the head on the perch.3. 336 .g. If there is a bacterial infection. However. is often localized on the head and near the ears (BOURDEAU 2004 personal communication). The most frequent bacteria isolated include Pseudomonas aeruginosa. mutans. Harvest mites of the family Trombiculidae may enter the ear. 4. Carcinoma of the ceruminous gland of the ear. The infraorbital sinus has rostral and caudal extensions near the ear.2. 4. grass lawn seed.1. The flea Echidnophaga gallinacea. Parasitic diseases Some parasites are seen near or in the ear of birds such as fly larvae. the tissues become hyperaemic and swollen and the opening is no longer clear.6. Foreign body It is possible to find foreign bodies in bird’s ear. Occlusion of the external openings may occur in macaws. Cnemidocoptes pilae or C. e. Enterobacter sp and Kocuria kristinae (DESMARCHELIER 2004 personal communication) a human opportunist bacteria. squamous cell carcinoma (KERN 1997) are the most common neoplasm and must be differentiated from chronic infection. Deafness is a possible sequel.35 days.1. with or without signs of vestibular dysfunction. Neoplasia Neoplasia can affect the ears. it must be opened surgically.1. Haemorrhage from the ear Trauma can occur and a haemorrhage in the external ear is usually due to a cranial injury.1. Infraorbital sinusitis can involve the ear. Acute or chronic bleeding from the external meatus may be present. Klebsiella sp.4. The ear opening may be swollen and red and the feathers may be matted with a discharge. There are disorders specific to the ear and other more generalised diseases that may affect the ears. It has been reported (KERN 1994) that the ear openings may also appear hyperaemic in birds with sinusitis. hypoboscis. Macaws hatch with a thin membrane covering their ear canals. If it does not.1. This membrane should start to open in 12 . Specific auricular diseases 4.1.1. 4. Otitis externa Otitis externa is an inflammation of the external ear. 4. squamous cell cancers are much more aggressive than basal cell cancers. It may by caused by bacterial or fungal organisms. 4. Lovebirds seem to be more prone to external ear infections (otitis externa) than other species of birds. A serous to purulent discharge may be noted.4 AURICULAR DISEASES Ear problems are not very common in birds. 4. • Discharge. opisthotonus.1. Pseudomonas aeruginosa infection was reported in an African grey parrot (Psittacus erithacus). feet and around the opening to the external ear 5. (2001). Chronic Pasteurella multocida infection is a frequent cause of otitis media and interna in poultry (KERN 1997). e. Otitis media and interna may result from septicaemia. Balance problem Birds with balance problems and regurgitate after rubbing on their toy.9 mm telescope is often needed to explore the deeper aspects of the canal. • Bleeding. Viral agents such as paramyxovirus (PMV) can cause otitis interna. may have ear infections. In pigeons. Fibroepithelial hyperplasia is a rare lesion of the external acoustic meatus.8. Ostriches (Struthio camelus) can develop poxvirus-induced lesions on the eyelids. and a mass protruding from the left ear. Neurological signs may range from depression. crusty lesions extending around eyes. (2001). resulting in loss of balance and torticollis. described a mycobacterial infection in a pionus parrot (Pionus menstruus) with weight loss. This loss was present with thickened. Non specific diseases TELL et al. and it is important to note if any of these signs are present: • Pruritus. lethargy. ulcers or proliferative (excessive growth) tissue. unilateral or bilateral paralysis of the legs and wings. • Nodular masses.7. ABRAMS et al.0 to 15. more than in the ear. Vascular or cardiac disease with associated anaemia. ataxia. hyperexcitability. Clinical signs associated with PMV infection in companion avian species are dependent on the strain of virus and the age. A 1. over the top of the head and invading the ear. • Large growths filling the ear canal. • Crusts. cerebrum. • Vestibular (balance) signs. PMV1 paratyphoid or polyneuritis. • Odour (rare).0 mm in passerine and psittacine birds) to very large. We have seen this in a PBFD positive kakariki (Cyanoramphus sp). In birds. brain stem and spinal cord. have also been incriminated. • Skin changes around the ear. described an Aspergillus blepharitis and dermatitis in a falcon with generalized feather loss. ears are hidden from 337 . a condition loosely described as vertigo has been reported quite commonly and whilst this is a non-specific term which may be applied to a variety of pathologies. circling and seizures. species and condition of the bird affected. 4. The opening is usually rounded but can vary in diameter from small (2. torticollis. CLINICAL EXAMINATION AND DIAGNOSIS Early in the illness. face. many birds have no obvious clinical signs. the possibility of otitis should not be overlooked nor should the ears be overlooked during post-mortem examination.1. Histopathologic lesions from PMV within the central nervous system are usually noted in the cerebellum.g. No particular problem has been encountered in our experience. Immunosupression is frequently associated with otitis and a PBFD PCR should be performed. A haemogram and skull radiography is necessary when the bird has clinical signs of otitis media or interna. To find the ears. Peripheral vestibular signs are often restricted to horizontal or rotatory nystagmus that does not change with changes in the position of the head. cranial nerve VIII. submit a biopsy sample for histopathology. A sample should be taken for bacterial and fungal culture and sensitivity testing. Falconidae and Strigiformes. The vestibulocochlear nerve. dorsoventral and lateral projection are good while in the others dorsoventral projection is not helpful and lateral projection is of limited use. Vertical nystagmus is associated with brain stem lesions. 338 . Electrosurgery is the preferred technique. provides information on the threedimensional position of the head in space as well as hearing (loss of function would result in deafness.1. An antibiotic or antifungal solution can be applied two to three times daily into the ear canal. Corticosteroids are given topically and not systemically. Usually these animals are more difficult to rouse. postural reaction deficits are on the ipsilateral side of the lesions. head tilt and/or nystagmus.view. Consider broad spectrum systemic antibiotic therapy for suspected bacterial otitis media and interna and provide appropriate supportive care. KRAUTWALD et al. Central disease involves the brainstem and carries a poorer prognosis than peripheral disease. If a mass is present. In the Psittaciformes.2. Corticosteroids may also be helpful in reducing inflammation in the external part of the ear. and/or loss of balance). Ophthalmic antimicrobial preparations without steroids are convenient to use. occlusion of the external openings. Cultures and cytology may also be necessary to confirm a diagnosis. or neoplasia. 6. Cochlear function is tested by examining the patient’s response to a sudden loud noise. The ear canal should be swabbed for cytology and Gram’s staining of the exudate. look behind and below the eye. Possibly other cranial nerve deficits may be present. The feathers directly over the ears have a slightly different texture and appearance. Surgical Surgery is necessary for foreign body removal. With vestibular disease. (1998) had shown that the suitability of the radiographic projections to demonstrate the anatomic structure of the external auditory meatus is different in the Psittacifomes compared to the Accipitridae. radiography or endoscopy. Medical Treatment is by cleaning and the application of topical antibiotics. it is important to determine whether the lesion is central or peripheral. 6 TREATMENT 6. circling. Part them and the ears will be apparent. The clinical examination consists of a visual inspection. Central lesions are often associated with changes in mental status. 589. WOODS L and CROMIE RL. DORRESTEIN GM and QUESENBERRY K. 192 Avenue de Romans. Lake Worth: Winger Publishing Inc 1994: 144 – 175. Psittacine Neonatal Development. CLUBB SL. The Avian ear and
[email protected]. TELL LA. 8 CITATION INDEX 1.120. Mycobacteriosis in birds. In: ALTMAN RB. HARRISON GJ and RITCHIE. 2. ABRAMS et al. DOOLING RJ and DENT ML. CES d’Ophtalmologie Clinique vétérinaire de l’Arche. 5. 583 .7 CONCLUSION Auricular diseases are uncommon in birds but the avian veterinarian must include the ear in the clinical examination. J Acoustical Soc Japan 2001. 26000 Valence. In: Whittow GC (ed): Sturkies’s Avian Physiology.fr 339 . 7. Winged Wisdom. Consider that immunosupression is frequently associated with otitis and perform a PCR for diagnosis of PBFD. France Email: franck. J Avian Med Surg 1998. In: HARRISON GJ. NECKER R. Pet Bird Magazine. The avian ear: part I and II.June 2003 8. May . 20(1): 180 .100. 2001. (ed): Avian Medicine and Surgery. San Diego: Academic Press 2000. 21 . 3. KRAUTWALD-JUNGHANNS ME. Disorders of the special senses. HARRISON LR and RITCHIE BW (ed) Avian Medicine: principles and application. Techniques and Research. Psittacine Aviculture. KERN TJ. ABRC Publications 1992. PESEK L. Philadelphia: WB Saunders 1997. Making distinctions in the physical examination. 12 – 17.203 AUTHORS ADDRESS F. the ear. CLUBB KJ and CLUBB SL. 6. 5th edition. 15(2): 114 . 9. Rival. In: Rev Sci Tech Off Int Epiz. SCHUBOT M. Aspergillus blepharitis and dermatitis in a peregrine falcongyr falcon hybrid. DVM. New studies on hair cell regeneration in birds.157. Perspectives. Président du GENAC. 4. 12 (3): 149 . J Avian Med Surg 2001. KOSTKA VM and DÖRSCH B. Comparative studies on the diagnostic value of conventional radiography and computed tomography in evaluating the heads of psittacine and raptorial birds.. 22: 93 . vigour.Oxidative stress . The future fecundity of female nestlings has also been linked to the nutrition received both from the egg and immediately post-hatch 340 .Antioxidants ABSTRACT The success of the avian reproductive cycle from the induction of follicle formation through ovulation. Stockdale BVM&S. early feeding behaviour. egg production. England United Kingdom THE NUTRITIONAL IMPLICATIONS OF PRODUCING THE ‘OPTIMAL EGG’ IN CAPTIVE BIRDS B. white. The egg composition is designed in such a way that all nutrients necessary for the development of the future embryo are accumulated within the egg yolk.Avian embryo . a heavy onus is placed on the nutritional status of the female bird. fertilization. MRCVS KEYWORDS Nutrients . This paper provides an overview of some of the nutritional dynamics involved in the production of the ‘optimal egg’. the egg.Dead-in-shell . and the immune status of the chick. Because all the nutrients required for embryonic development and survival must be placed within the egg prior to shell formation. Poor maternal state at the start of breeding affects the female bird’s ability to produce and incubate eggs. 1 INTRODUCTION Avian embryonic development takes place in a semi-closed environment. and recycle. a nutrient complete diet both pre and during the ovulatory period are essential in ensuring that the requisite essential nutritional components are available for optimal egg formation. Leicestershire.Meadow Lane Veterinary Centre. Poor egg quality affects chick hatch size. Loughborough. The implications of inadequate egg nutrition are reviewed and it is proposed that nutrition in captive birds may play a more prominent role in reproductive failures. and the subsequent rearing of the young. It also influences their willingness to provide parental care and ability to rear chicks up to fledging. and shell. C. characterised by deadin-shell than is currently appreciated. is dependent on adequate and appropriate nutrition. As egg nutrient provision is funded from both endogenous protein reserves and exogenous dietary sources. where only exchanges of gas and water take place. In most species. development of the oviduct and accretion of egg proteins. lipids. from nutrients stored in anticipation of egg synthesis. The growth of the oviduct and the synthesis of several yolks are mostly complete before the first egg is laid. the nutritional needs of laying birds include the nutrients required for general maintenance. The requirements for specific essential amino acids for egg production. and cysteine are commonly cited as limiting amino acids. showing how dietary manipulation can achieve higher production in terms of fertility. Birds laying eggs require protein for maintenance. Maternal nutrition can therefore be considered as the major determinant in the development and health of the avian embryo and viability in early post hatch life. the female’s requirements increase at least a week prior to her first oviposition.1 times greater than maintenance being required for egg production in the budgerigar. i. egg albumen is synthesised in the oviduct during a 24h period before ovulation. in proportion to the birds’ needs. from nutrient reserves. are often far in excess of those required for maintenance.e. this should not detract from the hen’s worth as a model for understanding the consequences of inadequate embryo nutrition in other avian species. methionine. The additional nutritional needs may be met either directly from the diet.1. 2 OVERVIEW During reproduction. or any combination of the above (MURPHY 1994). however. which may include stores or any tissue component that can be drawn upon to reallocate endogenous nutrients when daily intake falls below needs. Lysine. These estimates assume that the dietary protein is of satisfactory quality. Estimates of the additional protein requirements of several avian groups range from 86-232% of their minimum maintenance requirements (ROBBINS 1981). a rapid increase in tissue mass of the female reproductive tract. and weight gain in domestic poultry. and overinterpreting the parallels between precocial chicks and altricial nestlings.(GORMAN and NAGER 2004). levels of lysine 4. Consequently. Whilst care must be observed when discussing levels of specific nutrients. postnatal survival. and antioxidants are specifically indicated and the following sections highlight their respective roles in the production of the ‘optimal egg’. proteins. however. that the pattern of the amino acids in the dietary protein is supplied more or less. 2. The quality of these proteins is reflected by the quality of the daily dietary protein levels (MURPHY 1994). Among different nutrients that can significantly affect chick embryo development and subsequent viability. thus dietary amino acid requirements are especially high on the day preceding each oviposition. hatchability. Very little work has been done on the specific role of nutrition in achieving improved reproductive performance in birds kept as avicultural specimens. A wealth of literature does exist. for example. The biochemical and biological nature of the deficiencies and excesses within the egg should still hold true. 341 . Protein as a limiting factor Egg production is a protein-demanding process. with. and those required for egg synthesis. 2. lipids. The importance of lipoproteins for chick embryo development is unequivocal (SURI 2002). Lipids and polyunsaturated fatty acids (PUFAs) The production of a clutch of eggs requires the deposition of large amounts of yolk lipids. suggesting that the use of tissue protein to assist with egg production may be widespread in birds. has been shown to affect embryonic development. as well as providing an energy supply. mostly during the several days prior to ovulation. 342 . not in proportion to the needs for synthesis. 2. The individual amino acids are released more or less in proportion to their occurrence in proteins breaking down. body plundering occurs and use is made of endogenous reserves. on the levels of amino acids she is able to fund for the next clutch of eggs and her ability to both incubate and rear her nestlings. even if they restore tissue proteins the next day.Where dietary protein levels fall below the required levels for reproductive performance. This can have a serious knock-on effect on the overall health of the female. Dietary requirements of essential fatty acids The fatty acid composition of the maternal diet and subsequent inappropriate fatty acid provision within the yolk. A significant loss of muscle weight at the time of egg formation is documented in 21 of 29 species studied (HOUSTON 1995). protein removal from the sarcoplasm occurs over a range of different proteins. Deposition of egg yolk is completed 24h before its ovulation. and is recognised as a cause of increased chick mortality (VILCHEZ 1990). is sufficient for the adequate maintenance of the chick for several days post hatch. viability. Overnight. Yolk lipids and proteins are synthesised in the liver under the influence of oestrogen and progesterone and are transferred through the blood to the ovarian follicles. and hatchability. All tissues need to be considered as potential protein sources. During embryonic development. Eggs are enriched with methionine and cysteine relative to skeletal muscle and so greater volumes of endogenous proteins must be used to provide for those proteins in the egg. Lipids in the yolk are of two main types: specific yolk-targeted very low-density lipoproteins (VLDLy) and vitellogenins (VTN).3. Lipids comprise about 10% of the total weight of an egg and 33% of the yolk weight. The importance of PUFAs in cell metabolism is difficult to overestimate (SURI 2002). but the pectoral muscle mass provides the main supply. in the form of residual yolk. During egg laying. all birds rely at least partially on amino acids derived from tissue proteins to synthesize egg proteins. This is a costly process for the female bird as the balance of amino acids in skeletal muscle is sufficiently different from that in egg protein. Approximately 80% of the entire lipid content of the yolk is mobilised and absorbed into the embryonic tissues over the last few days of development and the remainder. About 90% of the energy required for the developing chick is supplied from egg yolk by fatty acid oxidation. play a crucial role in providing the developing embryo with a source of important constituents for the production of biological membranes and biologically active substances.2. Almost all the lipids in the yolk are present as lipoprotein complexes with an overall lipid:protein ratio of about 2:1. As well as having sufficient levels of essential fatty acids in the maternal diet. the water soluble antioxidants. Vitamin E. Little work has been done other than with gallinaceous birds into the ability of avian species to convert these fatty acids and it is suggested that given the variability among other vertebrate taxa. it is important that they are supplied in approximately equal proportions. vitamin E and carotenoids. The main fat-soluble antioxidants. In the chicken it has been shown that producing each egg requires the release of twice as much vitamin E as is stored in the liver (SURI 2002) indicating that the liver is not a good reservoir of vitamin E in the laying hen. LNA is the parent for n-3 fatty acids and LA the parent for the n-6 series of fatty acids and they are not inter-convertible. The antioxidant system of the newly hatched chick includes the fat-soluble antioxidants. linseed and rape oils are good sources of LNA and grains and plant oils such as corn.5.The basic building block fatty acids that are deemed to be essential dietary requirements for birds are linoleic (LA) and linolenic (LNA) acids. the embryonic liver contains high levels of PUFAs derived from the yolk that require a considerable degree of antioxidant defence against peroxidation. It is considered to play a central role in antioxidant production during embryogenesis and low levels have been associated with decreased hatchability and slow embryonic development. Yolk lipid droplets containing vitamin E are taken up by the yolk sac membrane and processed into lipoprotein particles that are released into the embryo circulation. At hatching. ascorbic acid and glutathione.4. Vitamin E is a generic term which encompasses eight main tocol and tocotrienol derivatives (SURI 2002). 2. Green leafy vegetables. These lipoproteins 343 . so must be derived from the diet. Both of these groups compete for the same modifying enzyme systems and a preponderance of one affects the levels of the other. and sunflower are rich sources of LA. Antioxidant systems and avian embryo development The animal body is under constant attack from free radicals formed as a natural consequence of the body’s normal metabolic activity within its tissues and antioxidants provide protection against their damaging effects. Failure to prevent lipid peroxidation within the egg will potentially result in excessive tissue damage and embryonic death. 2. as well as antioxidant enzymes. The main site of vitamin E accumulation in the avian embryo is the liver. that diets for birds should also contain arachidonic acid and eicosapentanoic acid (KLASING 1998). and preservation of their integrity and function. cannot be synthesised de novo by animals. Vitamin E is transported from the maternal diet to the hen bird’s liver and further to the developing oocyte as a component of VLDLy and is subsequently deposited within the lipid rich fraction of the yolk. Exclusion of vitamin E from the diet can therefore cause a rapid decrease in tocopherol concentration in the egg yolk. vitamin E and carotenoids. The most important effect of free radicals on the cellular metabolism is in their participation in lipid peroxidation. Natural foods contain differing proportions of these fatty acids. Prime targets for peroxidation are the PUFAs that are integral in the structure of cellular membranes. Although fat deposits contain a high level of carotenoid pigments. This can result in high levels of ‘pipping death’. surprisingly.are then metabolised by various tissue lipoprotein lipases and the resultant remnants taken up by the embryo liver. They also function as important visual indicators of offspring health. plays a crucial role in maintaining physiological levels of vitamin E in the chicken tissue during the first ten days of postnatal development. This may be explained as a result of low pancreatic lipase activity and restricted bile production in the developing avian alimentary tract. but species specificity in absorption and transfer to the egg would seem to occur. indicating that dietary intake is insufficient to maintain reserves. they are considered to be ‘sinks’ rather than stores with the carotenoids being unavailable for use except during fat metabolism. This is probably a direct effect of dietary manipulations . carotenoids are deposited in a dose dependant manner. the toe web. therefore. as opposed to a set level deposition. Those wild birds studied having a much higher egg yolk carotenoid level than those commercially reared. Maternal diet. The main storage sites within the body are the liver and. The intensity of the colour is thought to be a true reflection of the quality of the nestling’s health . can be explained by the mobilization of carotenoid stores for incorporation into the yolk. and the levels in eggs respond very quickly to dietary level changes.pale gapes being a consequence of insufficient provision or diversion of carotenoids to combat disease. The gradual loss of pigment from the shank and beak of certain birds as egg production continues. which occur in varying levels. Liver levels of vitamin E in chickens drop dramatically over the first few weeks post-hatching. The concentration of tocopherol in the liver tissue increases 3-5 times in the last week (chicken) of incubation reaching a maximum just after hatching. The passerine nestling also shows a similar lack of enzyme activity (CAVIEDES-VIDAL 2001). zeaxanthin and carotene. Vitamin E is particularly important at hatching.6. Carotenoids Carotenoids play a specific role in avian embryonic development and there are some indications that carotenoids are extremely important elements in maintaining the chick immune system. Absorption follows a similar pattern to that of vitamin E. The bright gape flanges of nestlings act as a temporal stimulus eliciting parental care and feeding.access to a wider range of variable carotenoid containing 344 . The most common carotenoids in seeds and plants are lutein. it is possible that lipoproteins are released directly into the portal vein and transported straight to the liver. imparting its characteristic yellow colouration. Plasma levels vary significantly depending on dietary provision and assimilation. when the onset of pulmonary respiration results in increased oxygen tension and increased free radical production. 2. During the rapid growth of the ova. Studies are limited. digestion of the food matrix is followed by the formation of lipid mixed micelles which are taken up into intestinal mucosal cells and delivered to the plasma via lymph system. As the lymphatics are not well developed in birds. It may also be relevant that the antioxidant activity of carotenoids is mainly expressed at the low oxygen tensions that prevail in embryonic tissues. food availability is a much stronger cue. This increases to over 23% at breeding without a substantial increase in energy requirements. Small birds appear. (a move in breeding granivorous birds to also consume high protein legume seeds and invertebrates). with no additional provision for the nutritional demands of egg laying. to have limited ability to actively exploit dietary amino acid complementation. The provision of a range of foods whilst offering a wider amino acid profile may not necessarily correct the problem. particularly small passerines. some of which lay a clutch of eggs weighing more than the female’s own body weight. Whilst mobilization of body tissues occurs in many species to aid with the 345 . As reproduction is a life-compromising activity there is inevitably a conflict between female and egg. or by choosing foods in amounts that permit complementation of constituent amino acids. Foraging birds can find foods containing protein of various quality to satisfy their amino acid needs by either choosing only those foods that contain a suitable array of essential amino acids – protein shift.5%. as energy expenditure and thus demand. but may indicate a more prominent role for carotenoids in embryo protection in certain species. Whilst the proximate factor in initiating breeding in birds from temperate regions is photoperiodic change. Reviews of captive bird diets focus principally on maintenance levels of nutrients and generally fail to address the issue of additional reproductive nutritional needs. the deficiency is in quality rather than quantity. dry seeds still make up the main ingredients of many captive bird diets. The protein component of ripe cereal seeds has long been recognized as having the potential to be nutritionally limiting for birds. Conservative diets aimed at providing maintenance levels of nutrients. When protein is the limiting factor in cereal-based diets. These tend to be misleading. however. Reproduction puts an even greater strain on a limited amino acid pool. for example. the results of a poor match between the amino acid balance of seed and the requirements for synthesis and renewal of tissue proteins. require a maintenance level of protein around 5. Feeding levels for essential nutrients (as far as these are known) are usually linked to energy based formulae. These have a much higher essential amino acid percentage than dried seeds (ALLEN and HUME 1997). in captive birds is much lower than wild birds. for opportunistic breeders from neotropical regions (where a large number of our avicultural species originate). Nevertheless. Japanese quail (Coturnix coturnix japonica). Biological mechanisms exist within wild birds to time breeding to coincide with the production of ripening seeds. 3 DISCUSSION The production of eggs is a nutritionally demanding process for many birds. Their dietary habits are fixed more by their mechanical ability both to husk and consume a seed type in the most expedient time. regardless of content. Protein levels geared to energy requirements of wild birds will grossly under-fund captive birds protein needs.food substances. may be contributing to reduced reproductive potential in captive birds. As more evidence accumulates showing that the maternal diet can have a profound effect on the health status. to a greater extent. It is suggested that low-quality neonatal nutrition results in a long-term impairment in the capacity to assimilate dietary antioxidants. Austr J Ecology 1997. Captive birds are limited by the very nature of their captive diets. the relative costs are high. Less is known about the features of vitamin E metabolism and accumulation in the egg yolk of wild birds. dead in shell. In the zebra finch. on endogenous reserves. perhaps it is time that we reassessed the role of nutrition in reproductive management of captive birds. sufficient carry over from diet to egg on a daily basis must be made in breeding birds. 346 .418. Artificially high dietary vitamin A levels can equally have an adverse effect on vitamin E levels. in their ability to manipulate their protein intake. 74: 769 . The importance of green seed in the nitrogen nutrition of the Zebra Finch. over 70% of the female’s daily intake of tocopherol and carotenoids are deposited in the egg yolk. Whether by preferential selection or chance consumption. and nestling desertion may all have a greater nutritional basis than currently appreciated. As vitamin E is poorly stored. especially psittacine birds. therefore. ALLEN LR and HUME ID. Independent of later nutrition. 22: 412 . individuals experiencing a short period of low quality nutrition during the nestling period have a two-fold reduction of these antioxidants at adulthood. resulting in a reduction of both reproductive and actual longevity. failure to incubate. can be. 2. Adequate provision of antioxidants is essential for successful embryonic development and hatching. Diets for breeding birds need to better reflect their reproductive needs. to accommodate the provisioning of the egg. CAVIDES-VIDEL E and KARASOV WH. Food quantity should never be an issue in captive bird management. therefore. A lot of attention has been focused on vitamin A levels in avian diets but little on the levels of carotenoids. 4 CITATION INDEX 1. Physiol Biochem Zoology 2001. nestling mortality. The deposition of these antioxidants declines with laying sequence. suggesting that the decline in provisioning may be a consequence of resource limitation (SURI 2002).amino acid needs of reproduction. Failure to breed. Supplementation of diets with foods high in limiting amino acids or the use of formulated diets may achieve a solution. expressed in terms of available nutrients. To date there is little information available relating to the tocopherol content of the egg yolk of different avian species.782. Low dietary vitamin A can have a direct adverse effect on reproduction and a knockon effect on the levels of carotenoids availability. Developmental changes in digestive physiology of nestling house sparrows. growth and development of the avian embryo and newly hatched chick. balance their amino acid intake. wild birds are able. the level of vitamin E in the diet will have a direct influence. vitamin A levels may also be marginal. More onus is placed. With many avicultural species. Species-specific differences in the metabolism of vitamin E may influence yolk levels but perhaps more importantly for captive birds. whereas food quality. Listed maintenance levels for vitamin E need to be increased. Nottingham Press. 9 Meadow Lane. 25: 27-38.1928. Condor 1981. ROBBINS C. 9.1538 AUTHORS ADDRESS Brian C Stockdale BVM&S MRCVS Meadow Lane Veterinary Centre. Estimation of the relative protein costs of reproduction in birds.co. 5. Natural antioxidants in avian nutrition and reproduction. LE11 1JU. CAB International 1998: 171172. KLASING K. SURI P. 83: 177. Comparative avian nutrition. Changes in the muscle condition of female zebra finches during egg laying and the role of protein storage in bird skeletal muscle.pipex. 8. Ibis 1995. HOUSTON DC et al. GORMAN H and NAGER R. 2002 VILCHEZ C et al. England Email: the. The influence of supplemental corn oil and free fatty acids on the reproductive performance of Japanese quail (Coturnix japonica).uk 347 .stockdales@dial. Poultry Sci 1990. 1923 . 7. Amino acid composition of avian eggs and tissue: Nutritional implications. 6. Proc R Soc London B 2004. Leicestershire. MURPHY M.T. J Avian Biol 1994. 69: 1533 . 137: 322 .328.3. Prenatal developmental conditions have long-term effects on offspring fecundity. 4. Loughborough. These diets also contain high levels of phosphorus that form phylate complexes with calcium reducing the bio availability of the mineral. Hypocalcaemia is a commonly recognised syndrome in grey parrots in captivity although the aetiology is still unconfirmed (ROSSKOPF et. Two groups of grey parrots were fed different diets for 12 months whilst being exposed to UV-B radiation from fluorescent tubes. In captive bred grey parrots osteodystrophy is a common clinical sign in young birds with deformity of the long bones easily identified radiographically (HARCOURT-BROWN 2003).Ionised Calcium ABSTRACT This study investigates the effects of exposing healthy captive grey parrots to artificial UV-B (315-285nm) radiation for 12 hours daily. Seed based diets are commonly fed to grey parrots in captivity containing low levels of calcium and vitamin D3. which respond to calcium or vitamin D therapy (HOCHLEITHNER 1989). erithacus) is wide spread throughout equatorial Africa. al. It is thought that the syndrome could be due either to a primary hypoparathyroidism or a secondary hyperparathyroidism due to inadequate husbandry. ranging from slight ataxia to seizures. Stanford BVSc MRCVS KEYWORDS Grey parrot – Ultraviolet radiation – Hypocalcaemia – Vitamin D. A significant increase in the serum concentration of 25 hydroxycholecalciferol and ionised calcium was found independent of the diet fed after UV-B radiation exposure. 1 INTRODUCTION The grey parrot (Psittacus e. Affected adult birds present clinically with a variety of neurological signs. 1985). It is the second most commonly traded psittacine bird in the world pet trade desired mainly due to its excellent ability to mimic (DEL HOYO 1997).Birch Heath Veterinary Clinic Cheshire United Kingdom THE EFFECT OF UV-B RADIATION ON CALCIUM METABOLISM IN GREY PARROTS M. Captive breeding of the grey parrot has increased dramatically in the last decade and it has become a common patient for veterinary surgeons worldwide. It has therefore been 348 . Failure to provide adequate UV-B light in poultry kept indoors will produce symptoms of vitamin D3 deficiency. 1994). The role of UV-B (285-315nm) radiation in the control of vitamin D metabolism has not been researched in psittacine birds at the present time. The skin has been established as the organ for vitamin D3 production as in mammals. 1941). After translocation into the circulation vitamin D3 is transported bound to a specific globulin binding protein. The measurement of 25 hydroxycholecalciferol has been shown to be a useful indicator of the UV-B exposure of an individual. rarely develop clinical signs of hypocalcaemia (RANDELL 1981). 1994). Vitamin D is supplied to birds from a combination of endogenous synthesis and dietary supply. Cholecalciferol is initially metabolised to 25 hydroxycholecalciferol in the liver (BLUNT et al. 1982). 1968). Provitamin D is converted to cholecalciferol (Previtamin D3) by an ultraviolet light (285-315nm wavelength) dependent isomerisation reaction. the active metabolites of cholecalciferol in the domestic fowl. The most significant active metabolite of vitamin D3 in domestic chickens is 1. In captivity grey parrots are usually kept indoors with limited access to ultraviolet light. Once sufficient previtamin D3 has been formed additional solar radiation transforms the provitamin D to biologically inactive compounds lumisterol and tachysterol (HOLICK 1994). 25 dihydroxycholecalciferol. 25 dihydroxycholecalciferol controlling both bone development and intestinal calcium absorption. 25 dihydroxycholecalciferol or 24.3nm wavelength (HOLICK et al. Recently it has been shown that there is thirty times more Provitamin D3 on the featherless leg skin than the back indicating the importance of this area for vitamin D3 metabolism (TIAN et al. 1989). The maximum conversion of provitamin D to Previtamin D occurs at the 297+/. The ultraviolet light required for endogenous vitamin D synthesis can either be supplied naturally from full spectrum sunlight or using artificial lamps manufactured to provide UV-B radiation. In one study none of the commercial lamps produced significant amounts of UVB wavelength in the vitamin synthesis spectrum 349 . Cholecalciferol can be stored in adipose tissue but to be physiologically active it must be metabolised by a 2-stage hydroxylation process (HOLICK 1995). Cholecalciferol is a sterol prohormone which undergoes a temperature dependent isomerisation reaction to form vitamin D3 (HOLICK. Birds secrete 7-dehydrocholesterol (Provitamin D3) onto the featherless areas of skin (Koch et al. This might suggest that they require higher ultraviolet light supplementation than other psittacine birds. This explains why hypervitaminosis D has never been reported from excessive exposure to UV-B light. Fluorescent tubes that provide some UVB radiation in addition to visible light are available commercially mainly designed for captive reptile exhibits (LOGAN 1969).postulated that poor diet contributes to a nutritional secondary hyperparathyroidism (Klasing 1998) but this does not explain why other psittacine species despite being fed the same diets. A lack of adequate ultraviolet light (UV-B spectrum 285-315nm) in captive birds may lead to a functional vitamin D deficiency and subsequent problems with calcium metabolism. There is a time delay between vitamin D3 production on the skin and its translocation into the circulation (TIAN et al. Studies have been conducted in poultry to demonstrate the requirement for cholecalciferol kept in the absence of ultraviolet light indicating (EDWARDS et al.. 25 hydroxycholecalciferol is transported to the kidney via carrier proteins and converted to either 1. 1994). If ultraviolet light was excluded and dietary cholecalciferol concentrations fell below 400ICU/kg symptoms of rickets were seen with concurrent low plasma ionised calcium concentrations. the other group was maintained on an unsupplemented mixed seed diet (Tidymix. After 12 months exposure to artificial UV-B lighting further blood tests were taken during the annual health examination to assess the effects of UV-B radiation on calcium metabolism in grey parrots. Oxford UK) was used to demonstrate an increase in the ultraviolet light levels experienced by all the birds in the third year. On the basis of these tests only healthy birds were included in the main population. Nebraska. Faecal samples taken from each individual had been subject to parasitology. Croydon. microbiology and gram stain examinations. Cairo New Road. US). Blood samples taken from each bird were subject to routine haematological and biochemical analysis including circovirus. The tubes were replaced after 6 months according to the manufacturers’ instructions. polyoma virus and chlamydophilia PCR tests. An Elsec UV-B light monitor 763 (Littlemoore Scientific Engineering. The monitor uses two photodiodes to detect UV-B radiation in the 285-400nm wavelengths expressed as mW/M2. UK). 40 birds were selected from the main population using a simple randomisation process to form the study group. Arcadia House. One group was fed Harrison’s High Potency Course pellet diet (Harrisons International Bird Foods. Two tubes were supplied to each pair of birds. John Heath. Railway Lane.5 metres from the tubes. The light was supplied using 1200mm 36W FB36 Arcadia bird lamps (Arcadia. Each aviary had a wooden nest box measuring 40cm by 30cm of shoebox design. When perching each birds would be a maximum distance of 0. The aim of this study was to investigate the effects of artificially supplied UV-B radiation on 2 groups of healthy grey parrots on different dietary regimes. A reflector (Arcadia ALR36) was mounted behind each tube to direct light towards the birds maximising the amount of UV-B that each bird received. The birds were kept in the same building under the same conditions as the main population. The group of 40 birds was randomly allocated into 2 groups of 10 pairs of grey parrots (n=20 birds per group: 10 male and 10 female). Each bird in the main population had been examined clinically prior to purchase in 1998 using a standard format. All the birds were purchased from a single source as wild caught imports from Guyana. Each bird was examined by laparoscopy to confirm sexual maturity and gender. 350 . All of the birds were subsequently placed under artificial ultraviolet light (UV-B 285315) for 12 hours daily whilst keeping the diets constant.so clearly clear should be taken in lamp selection (BERNARD 1995). After a year of no exposure to UV-B radiation during the annual health examination additional blood was taken from the study group to assay 25 hydroxycholecalciferol and ionised calcium in grey parrots with the informed consent of the owner. UK) suspended directly above each pair of birds. 2 MATERIALS AND METHODS The main population consisted of 100 healthy sexually mature grey parrots housed indoors as 50 pairs. All the aviaries were positioned in a single span farm building of brick and slate roof construction with no exposure to natural ultraviolet light. Each pair was housed in an identical individual aviary measuring 2m by 2m of identical brick and wire construction. Hull. 85 95% of mean 26.0 P value 0.20 71.77 139.24 SD 0.83 17.22 16.47 130.27 3.56 SE 21.23 1.01 0.23 69. 3 RESULTS 3.20-1.All the blood samples were taken under isoflurane anaesthesia.45 112.0 123.0038 0.71-116.2753 351 .20 1. The blood testing was performed on September 1st each year outside the breeding season to minimise the effects of both oestrogen and seasonality on both vitamin D3 and calcium levels (BENTLEY 1998).66 115.66 115.23 1.08-1.02 0.3 Effect of UV-B radiation on each dietary group Parameter Ionised calcium 25 hydroxycholecalciferol Dietary group Seed Pellet Seed Pellet Mean year 1 1.01 0.5 18.10 1.44 Wilcoxon W statistic 2.14 Median 35.31 4.55 106.01 Median 1.13 1.2 Descriptive statistics ionised calcium (mmol/l) Year of Study Year 1 Dietary Group Seed Pellet Seed Year 2 Pellet Mean 1.06 0.22-1.0053 0.94 102. A 2ml blood sample was taken from each bird split equally into heparin and EDTA eperndorph tubes.24 1.11 1.72-124.1 Descriptive statistics 25 hydroxycholecalciferol (nmol/l) Year of Study Dietary group Seed Year 1 Pellet Seed Year 2 Pellet Mean 71.25 1.77 Mean year 2 1.0 27.07 0.44 SD 90.0001 0.05 SE 0.01 108.23 78. The plasma sample was immediately cooled to –70 Celsius for subsequent analysis for parathyroid hormone.37 3.05 0.24 139.16-1.25 118.19 1.40 122.47 130.61-182.22 24. The EDTA sample was centrifuged for 15 minutes at 1600/s and the plasma titrated into another eperndorph tube.78-176.11 1.23 1.24 95% of mean 1. As most captive grey parrots are either kept indoors or live in Northern latitudes they would not be expected to receive similar ultraviolet light levels compared with equatorial Africa. 352 . This may well explain why grey parrots are so susceptible to symptoms of hypocalcaemia. Furthermore the birds are traditionally fed a diet with a low calcium and vitamin D content.66 115.24 139. At the beginning of the study there was a significant difference between the serum ionised calcium and vitamin D concentrations between the 2 dietary groups.6620 Kruskal-Wallis ANOVA statistic P value Ionised Calcium Vitamin D 4 DISCUSSION The study suggested that artificial UV-B radiation could be used in grey parrots to influence calcium metabolism.37 0. The author now recommends the provision of 2. The findings are potentially significant in a breed known to commonly suffer from disorders of calcium metabolism.3.4 Comparison between dietary groups after UV-B supplementation.5446 0.23 0. Parameter Dietary group Seed Pellet Seed Pellet Mean year 3 1. After the provision of UV-B light for 12 months there was no significant difference in 25 hydroxycholecalciferol or ionised calcium concentrations in the 2 dietary groups. Preferably this should come from solar radiation due to potential problems with supplying artificial UV-B with light both from the performance of the lamps and the practicalities of keeping the bulbs close to the birds. This was due to the higher vitamin D and calcium content in the pellet over the seed diet. The ionised calcium concentration was significantly increased in both the pellet and seed fed groups after the provision of UV-B radiation. This might suggest that the pellet fed group already had adequate stores of vitamin D in the form of 25 hydroxycholecalciferol.44 0.5% UV-B fluorescent lighting to all captive grey parrots kept indoors.19 1. The additional UV-B light would not lead to vitamin D toxicity due to the natural feedback mechanisms to prevent this. It is proposed that grey parrots should be provided with UV-B radiation as part of their husbandry. The vitamin D concentration was significantly increased in the seed fed group but not the pellet fed group. Future research should determine whether calcium metabolism in grey parrots differs from other species of psittacine birds concentrating particularly on the effect of ultraviolet light within different species. 35. K. 20: 33 .294. The hypocalcaemic syndrome in African Greys: An updated clinical viewpoint. KLASING. 73(2): 288 .C.W.295. Spectral irradiance of special fluorescent lamps and their efficiency for promoting vitamin D synthesis in herbivorous reptiles. 7: 3317 . and LANE R. P. 25 hydroxycholecalciferol. 5. 290 .3322 4. Michigan State University 1995. CW6 9UU United Kingdom Email:
[email protected] REFERENCES BENTLEY. AUTHORS ADDRESS Michael D Stanford BVSc MRCVS Birch Heath Veterinary Clinic Birch Heath Road. The full list of references is available from the author 1. DEL HOYO J. The provitamin of covering tissues of chickens. Poultry Science 1941. Convulsions in African Grey parrots in connection with hypocalcaemia: Five selected cases. DELUCA HF and SCHNOES HK.J. ELLIOT MA.A. eds: Handbook of the Birds of the World. Biochemistry 1968. Proc Assoc Avian Vet. 3. Incidence of juvenile osteodystrophy in hand reared parrots (Psittacus e psittacus). 2. 6. Sandgrouse to Cuckoos.132. 10. Cambridge: Cambridge University Press 1998. 152: 438 . A biologically active metabolite of vitamin D3. BERNARD JB. Cheshire. ROSSKOPF W.52. Hochleithner. HARCOURT-BROWN N.J. 44 . 1985. Comparative Avian Nutrition. BLUNT JW. WOERPEL R.. Comparative Vertebrate Endocrinology. KOCH EM and KOCH FC. 8. Quantitive requirement for cholecalciferol in the absence of ultraviolet light. 129 . Proc Euro Symp Avian Med Surg.com 353 . Tarporley. Poult Sci 1994. ELLIOT A and SARGATAL J.439. 7. PhD dissertation. M. New York: CAB International 1998. 269 . Vol 4.301. Vet Rec 2003. SOONCHARERYNING S and BRITTON WM. 9. EDWARDS HM JR. Barcelona: Lynx Editions 1997. 1989. The Animal Emergency Center1, Glendale, WI; Avian and Exotics2, Miami, FL; Angell Memorial Animal Hospital3, Boston, MA; Medical College of Wisconsin4, Milwaukee, WI; Avian and Wildlife Laboratories5, University of Miami, Miami, FL; Virginia Regional College of Veterinary Medicine6, United States of America RESPONSE TO FLUID RESUSCITATION AFTER ACUTE BLOOD LOSS IN MALLARD DUCKS (ANAS PLATYRHYNCHOS) M. Lichtenberger1, DVM, Dipl ACVECC, W. Chavez2, DVM, C. Cray5, PhD, C. Orcutt3, DVM, Dipl ABVP-Avian, D. DeBehnke4, MD, FACEP, E. Stumpp1, DVM, S. Diehl1, DVM, B. F. Feldman6, DVM, Ph.D, C. Page1, BSc, CVT, L. Mull1, BSc, CVT, L. Inman1, BSc, R. Kirby1, DVM, Dipl ACVIM, Dipl ACVECC KEYWORDS Hypovolemic shock – Resuscitation – Crystalloids - Colloids, Polychromasia ABSTRACT The pathophysiology of haemorrhagic shock is poorly understood in avian species. Acute blood loss of 30-40 % of blood volume has been shown to result in 50% mortality (LD50) in mammals. Blood loss is better tolerated in birds than in mammals. The LD50 for acute blood loss in ducks has been shown to be 60% of the total blood volume. No studies have been reported in birds documenting changes in heart rate and blood pressure after acute blood loss and fluid resuscitation. Reticulocytosis, or polychromasia, is the hallmark of intensified erythropoiesis, allowing classification of anaemias into regenerative or non-regenerative types. Reticulocyte release from the bone marrow in mammals commonly occurs 5 days after acute blood loss and rarely can occur as early as 2-4 days. No studies have been reported in birds after acute blood loss showing regenerative changes documented by absolute numbers of polychromasia, haematocrit and haemoglobin levels. The purpose of this study in mallard ducks (Anas platyrhynchos) was threefold: a) to demonstrate the heart rate and blood pressure response to acute blood loss at the predetermined LD50, b) to demonstrate if birds respond to acute blood loss with a baroreceptor response as seen in man and mammals and c) to document the efficacy and safety of smallvolume crystalloid resuscitation alone or in conjunction with colloids or Oxyglobin® after acute blood loss; and to demonstrate the time required for a regenerative red blood cell response after acute haemorrhage by documenting an increase in absolute polychromasia, haematocrit and haemoglobin concentrations. Twenty-seven mallard ducks were included in the study. Phlebotomy was performed on each duck to a LD50 of 60% of their total blood volume. The birds were randomized into three groups 354 to receive crystalloids alone, hetastarch and crystalloids, and Oxyglobin® and crystalloids. Indirect blood pressure measurements and heart rates were recorded throughout the study. The birds were recovered and observed for mortality over 96 hours. Blood was collected from 3 birds in each of the 3 fluid resuscitation groups at 0 h, 12 h, 24 h, 36 h, 48 h, 60 h, 72 h, 84 h and 96 h. The blood was evaluated for polychromatic changes, red blood cell count, haematocrit, and haemoglobin concentration. 1 INTRODUCTION Acute haemorrhagic shock is a syndrome resulting from reduction of the effective circulating volume, and in which impairment of the circulation steadily progresses until it eventuates into a state of irreversible circulatory failure (WIGGERS 1975). This definition of haemorrhagic shock was described a hundred years ago, but remains valid today with the outcome optimised by fluid resuscitation (MARINO 1997). The most common causes of blood loss in birds include traumatic injury and haemorrhagic lesions of internal organs, such as ulcerated neoplasms and proventricular/ventricular ulcerations (FELDMAN et al. 2000). Significant blood loss can occur during surgical procedures. Heavy infestation with blood-sucking ectoparasites or gastrointestinal parasites, coagulopathies associated with toxicities or severe liver disease are less common causes of blood loss anaemia in birds (FELDMAN et al. 2000). The concept of haemorrhagic shock is poorly understood in avian species. Acute blood loss of 30-40 % of blood volume has been shown to result in 50 % mortality (LD50) in mammals (MARINO 1997). Blood loss is tolerated better in birds than in mammals. The LD50 for acute blood loss in ducks has been shown to be 60% of their total blood volume (LICHTENBERGER et al. 2002). Acute blood loss of greater than 40 % of the blood volume in mammals usually marks the onset of hypovolemic shock with a subsequent increase in heart rate due to a strong baroreceptor response. A high level of sympathetic tone commonly occurs and is characterised by pale mucous membranes, prolonged capillary refill time, poor pulse quality and tachycardia. It has been stated that in the chicken, acute blood loss is not associated with a carotid sinus baroreceptor response as it is in mammals (JENKINS 1997, (STURKIE 1986). In mammals, optimal fluid resuscitation subsequent to acute blood loss results in a decrease in heart rate and increase in blood pressure (WIGGERS 1975). This study will investigate if birds have a similar baroreceptor response to shock as seen in mammals. No studies have been done in birds documenting changes in heart rate and blood pressure after acute blood loss and fluid resuscitation. When blood loss is severe, the priority of resuscitation is to expand the vascular space and improve cardiac output. Resuscitation implies an urgent need to restore tissue perfusion and oxygenation. The goal is to give the least amount of fluids to reach the desired endpoints of resuscitation. The endpoints of resuscitation are a normal blood pressure and heart rate. Crystalloids such as lactated Ringer’s solution are popular fluid therapy for shock as they are inexpensive, and readily available. However, in 355 shock resuscitation, only about 25% of the infused volume of crystalloids remains in the intravascular space by the end of infusion; therefore, it is of limited value as a plasma volume expander unless given repeatedly in very large volumes (TAKORI and SAFAR 1967). Large molecular weight colloids, such as hetastarch (HES) (Abbott Laboratories, North Chicago, IL, USA) and Oxyglobin® (Biopure Corporation, Cambridge, MA, USA), given via the intravenous or intraosseous route, remain primarily within the vascular space and do not freely pass into the interstitial space as do crystalloids. Thus colloids should be more effective than crystalloids for rapid, small-volume fluid resuscitation subsequent to haemorrhagic shock (MARINO 1997). After acute blood loss, mammals are dependent on red blood cell (RBC) regeneration to maintain oxygen delivery to the tissues. In response to tissue hypoxia, erythropoietin stimulates RBC production by the bone marrow. Reticulocyte release from the bone marrow in mammals occurs rarely after 2-4 days and most commonly longer than 5 days after acute blood loss (FELDMAN et al. 2000). Reticulocytosis, or polychromasia, is the hallmark of intensified erythropoiesis, allowing classification of anaemias into regenerative or non-regenerative types (FELDMAN et al. 2000). A previous study in acute blood loss in the duck documented a early regenerative response shown by presence of polychromasia at 36 hours after blood loss (LICHTENBERGER et al.2002). The relatively short life span of the red blood cell (28-45 days) and presence of a nucleated red blood cell (RBC) may account for a birds’ ability to mount a very early regenerative response. No studies have been done in birds with acute blood loss, showing regenerative changes documented by absolute numbers of polychromasia, changes in haematocrit (HCT), and haemoglobin (Hb) concentrations. The purpose of this study in mallard ducks (Anas platyrhynchos) was threefold: to demonstrate the heart rate and blood pressure response to acute blood loss (at the predetermined LD50) as well as the response to fluid resuscitation; to demonstrate the efficacy and safety of small-volume colloid resuscitation (HES and Oxyglobin®) with crystalloids as compared to small-volume crystalloid resuscitation after acute blood loss; and to demonstrate the time required for a regenerative red blood cell response after acute haemorrhage by documenting an increase in absolute polychromasia, HCT, and Hb concentrations. 2 MATERIALS AND METHODS Mallard ducks were used in the fluid administration studies. The fluid resuscitation studies were performed at the Medical College of Wisconsin research facility and were approved by the Medical College of Wisconsin Animal Care and Use Committee. All birds were housed at the Medical College of Wisconsin research facility during the study period and were fed a commercial pelleted ration. Nine of the 29 birds were housed in pairs in cages. The other 18 birds were housed together in large pens. Twenty nine mallard ducks (15 males and 15 females, less than 1 year of age) were initially included in this study. Physical examination findings and weight were recorded. Heart rates using an electrocardiogram (Escort, Medical Data Equipment, Arletta, CA, USA) and indirect blood pressure monitoring (Park Medical Electronics 356 Inc., Aloha, OR, USA) were recorded after anaesthesia induction and intubation, after phlebotomy, and after fluid resuscitation. A 19-gauge butterfly catheter was placed in the jugular vein, and 6 ml/100 gm body weight (determined as the LD50 for acute blood loss) was withdrawn over 10-20 minutes. Two birds were eliminated from the study, because they died after blood collection and prior to fluid resuscitation. Subsequently, the remaining 27 birds were randomized (9 birds in the crystalloid group, 9 birds in the HES/crystalloid group and 9 birds in the Oxyglobin®/crystalloid group) to receive either 15 ml/kg of an isotonic crystalloid, 10 ml/kg of a crystalloid with 5 ml/kg of HES, or 10 ml/kg of a crystalloid with 5 ml/kg of Oxyglobin® over a 5-minute period. Anaesthesia was turned off after the fluid resuscitation. Heart rate and blood pressure monitoring were recorded after fluid resuscitation and until the bird woke up from the anaesthesia. The birds were recovered in a cage. The birds were observed for mortality over the following 96 hours. Any surviving birds were then euthanised. Blood was collected from 3 birds in each of the 3 fluid resuscitation groups at 0 h (before phlebotomy, after phlebotomy and after fluid resuscitation), 12 h, 24 h, 36 h, 48 h, 60 h, 72 h, 84 h, and 96 h. A blood smear was made and the rest of the sample was placed in a microtainer (Microtainer Brand, Becton Dickinson Vacutainer Systems, Franklin Lakes, New Jersey, USA) with EDTA anticoagulant after a blood smear had been made. All blood smears and samples were sent to the Avian and Wildlife Laboratories (University of Miami, Miami, FL, USA) for evaluation within 24 hours from collection. The HCT was evaluated at 0 h, 24 h and every 12 hours until completion of the study. The haemoglobin concentrations were evaluated at 0 h, 48 h, and every 12 hours until completion of the study. The absolute degree of polychromasia (polychromatic cells/100 RBC cells with an average of 300 cells counted) was determined at 0 h and every 12 hours until completion of the study. Death rate between the three fluid resuscitation groups was statistically analysed using chi-square analysis. Polychromasia, RBC counts, haemoglobin (Hb), and HCT were evaluated using paired, 1 or 2 tailed T test. 3 RESULTS AND DISCUSSION Haemorrhagic shock is poorly understood and not well studied in avian species. During the fluid resuscitation study, there was a trend of decreased mortality in the Oxyglobin® group; however, we found no statistical difference in mortality rate among crystalloids, HES and Oxyglobin® used for resuscitation from acute blood loss. The oxyglobin group resuscitated quickly to normal haemodynamic parameters and awoke without signs of weakness. The HES and crystalloid group did not recover as well as the oxyglobin group. This may show that Oxyglobin® may be a superior resuscitation fluid for acute blood loss, when compared to HES or crystalloids. Further studies are needed to monitor the haemodynamic response to acute blood loss as well as to various protocols of fluid resuscitation in the avian species. Also, future studies need to be done using a larger number of birds and use of different volumes of fluids to a resuscitation endpoint. 357 All birds in the study became tachycardic with a decrease in blood pressure immediately post phlebotomy, and hypotensive and tachycardic when the amount exceeded 2030%. The heart rate and blood pressure returned to normal after fluid resuscitation in most of the birds. These responses may indicate that birds have a baroreceptor response to intravascular fluid volume changes as is seen in mammals. In this study, birds started to show a RBC regeneration starting at 12 hours after acute blood loss and peaking at 60 hours, as measured by the absolute number of polychromatic cells, Hb concentration and HCT. This response is much earlier than that seen in mammals. Further studies need to be done evaluating erythropoietin levels and bone marrow changes after acute blood loss. 4 CITATION INDEX 1. 2. 3. 4. 5. 6. 7. WIGGERS CI. Hemodynamic and metabolic alterations in peripheral tissue during haemorrhagic shock. Ann Surg 1975;11: 696 - 703. MARINO PL. Hemorrhage and hypovolemia. In: MARINO PL (ed): The ICU Book. Philadelphia, PA: Williams & Wilkins ; 1997; 207 - 227. FELDMAN BF, ZINKL JG and JAIN NC. Red blood cell regeneration. In: FELDMAN BF, ZINKL JG, JAIN NC (eds): Schalm’s Veterinary Hematology. Baltimore: Lippincott Williams & Wilkins; 2000; 445 - 467. LICHTENBERGER M, ORCUTT C, DeBEHNKE D, et al. Mortality and response to fluid resuscitation after acute blood loss in mallard ducks (Anas platyrhynnchos). Proc Assoc Avian Vet, Pittsburgh 2002: 65 - 70. JENKINS JH. Avian critical care and emergency medicine. In: ALTMAN AB, CLUBB SL, DORRESTEIN GM and QUESENBERRY K (eds): Avian Medicine and Surgery. Philadelphia: WB Saunders 1997: 839 - 863. STURKIE PD. Body fluids: Blood. In: STURKIE PD (ed): Avian Physiology. New York, NY: Springer-Verlag 1986: 102 - 121. TAKORI M and SAFAR P. Treatment of massive hemorrhage with colloid and crystalloid solutions. Studies in dogs. J Am Med. Assoc 1967; 199: 297. AUTHORS ADDRESS Marla Lichtenberger, DVM, DACVECC The Animal Emergency Center 2100 West Silver Spring Drive Glendale, WI, 53209 United States of America Email:
[email protected] 358 Clinic for Birds, Meppel, The Netherlands PARROTS DON’T BITE CHILDREN J. Hooimeijer, DVM CPBC KEYWORDS Parrots - Behaviour - Biting - Children ABSTRACT An impressive beak is one of the salient features of a parrot. It is also often the source of considerable anxiety for parents who associate a beak with biting and are concerned about the damage that could be inflicted on their children. It is therefore important to realize that a parrot does not use its beak in the wild in order to injure or kill, but for climbing, eating, preening, feeding and defending. 1 INTRODUCTION The beak of a parrot, a cockatoo or a macaw is an imposing instrument that many bird owners regard with a certain amount of awe and anxiety. The power behind a parrot’s beak is well known to everyone. In the wild, beaks are used to crack open hard nuts and strong seed coverings. Nesting holes in trees are enlarged using this same powerful tool. In captivity proffered tree branches are turned into matchsticks, nuts that have been fastened with a wrench are loosened from their bolts, and toys and furniture are reduced to fragments, all by these same beaks, and seemingly without effort. The amount of power that a parrot can exert with a lightweight skull and a lightweight beak is exceedingly impressive. By combining strong muscles and the hinge construction of the upper beak, parrot beaks can be as effective as a pair of strong sharp pliers. Apart from eating and nest building, the beak also has many other important nonaggressive functions. It is used as a third foot when the birds are climbing to keep them steady. It is used to hold objects so that the sensitive tongue can investigate them. The beak is also the instrument that is used to care for the bird’s own feathers and for those of his or her partner. Young birds are also cared for using the beak. 359 Biting is a frequently cited reason for getting rid of a pet parrot, who then disappears into the cycle of sale and re-sale, or is dumped in a rescue centre. The arrival of a baby in the house often coincides with the departure of the parrot because of the new parents’ fear that their offspring will not be safe around their pet. That beak, after all, what damage that could do to little fingers, little toes, little ears, or little nose!!! 2 BITING OTHER PARROTS IN THE WILD It is striking that there is no significant data to support the idea that parrots inflict serious or fatal bite wounds on each other in the wild. On the contrary, all evidence indicates that deliberately wounding or killing their fellows is not part of the natural behaviour of parrots. Although they are equipped with a built-in lethal weapon that could easily maim or kill another bird of their own kind, such behaviour is practically unknown in nature. Debilitating members of the same species is not in the interest of the preservation of that species. Skirmishes certainly take place, but these are mostly displays and mock fights in which real damage is seldom done. Parrots learn early to read the body language of their fellows and know precisely what is permitted and how far they can go in their combativeness. Playful romps with other youngsters are part of the learning and socialization process for every young parrot and seldom if ever lead to injury. 3 BITING OTHER PARROTS IN CAPTIVITY In captivity biting problems are seen most notably among the cockatoos, where males have been known to seriously injure or even kill a female. Pet cockatoos are also prone to express frustration by mutilating themselves using their beak. Neither of these behaviours is known to occur in the wild. Limitations due to the size of the housing of birds in captivity often hinder avoidance behaviour or make it impossible for the birds to respond appropriately to body language that in the wild would elicit a retreat from a confrontational situation. Unable to flee, a bird becomes insecure and aggressive. Attacking or biting other birds can be regarded as unnatural behaviour due to the circumstances of captivity. In captivity birds of some species have been known to attack and even kill sick or wounded fellows. Serious head wounds have been observed in budgies, cockatiels and lovebirds when they are housed in same-species groups. It is not uncommon that the dead birds are then cannibalised. The author is not aware to what extent, if at all, this occurs in nature. We can regard biting in captivity as an expression of insecurity, and thus part of a behaviour problem. We observe insecurity in birds that are not treated with respect, as well as in periods of hormonal or sexual activity, and in instances of physical problems or sickness. 360 Birds with a strong attachment to their owner exhibit bonding behaviour, which in turn causes territorial behaviour. This territoriality is often considered aggressive or dominant behaviour although it is actually insecure and defensive. Away from his own territory, or when the partner/owner is absent, the bird behaves completely differently. 4 BITING PEOPLE IN CAPTIVITY There is a constant stream of stories and anecdotes from parrot owners who report having been bitten by their pets. The number of instances where subsequent medical attention was necessary, however, remains exceedingly small. This is surprising considering the amount of damage a parrot beak could do if actually used with the intention to maim or injure. It is even more surprising that at the Clinic for Birds in the last 22 years we have not seen a single incident of a parrot biting a child. In addition to that, consistent inquiry by this author as to personal, anecdotal or media-covered experience of bitten children has not uncovered a single incident of a child being bitten by a parrot. Again, considering the actual capabilities of a parrot beak, and the size of a child’s finger, nose, or ear, one might have expected to hear stories of severed or mutilated young appendages. In our experience, parrots react completely differently to children compared to the way they react to adults. Apparently parrots view children in much the same way as human adults do: they are not seen as threatening and therefore do not make the birds feel insecure. This is all the more striking when compared with the behaviour of dogs. When a dog owner is afraid that his pet might bite a child, his insecurity turns the child into a confusing factor in the dog’s environment and increases the chances that the dog indeed will bite. In spite of the fact that most parents feel anxiety about the perceived risk that a parrot will bite a child, parrots do not respond to this by biting. At most, the bird plays a game in which he pretends to bite, but does not carry out the “threat”. The frightened reaction of the parents can be regarded as a “reward” for this undesired behaviour, thus reinforcing it. Even in situations where one could think that the parrot had every reason to bite, as when a child “pets” too hard, pulls a tail, or intentionally or inadvertently teases, parrots do not inflict the expected wounds. At most, a blackand-blue mark may be the result, and this is most often caused by pulling back of the finger or hand that was being “held” in the beak. Apparently parrots have a “natural” inhibition when it comes to biting children. This is all the more reason to respect their natural behaviour. In spite of the many mistakes that are made in handling them, biting is not a normal parrot behaviour pattern. 5 CONCLUSION Biting can be regarded as learned behaviour in parrots, behaviour that is often unintentionally rewarded. Parrots that bite are usually not approached and treated with respect. Allowing a parrot to see that a person is afraid of it, is a sign of disrespect. When a parrot sees that the owner has a problem, it becomes insecure and loses 361 its respect for the owner. It is striking to observe that children apparently cause no feelings of insecurity in parrots even when the parents make it clear that they do not trust the bird. There is every reason to have much respect for the intelligence and the normal behaviour of parrots. This is certainly the case when we realize that they adapt to extremely unnatural circumstances in order to integrate in captivity as house pets. It is even more impressive when we realize that parrots don’t bite children in spite of the fact that all too many children are victims of child abuse by their own parents or are victims of biting dogs. 6 ACKNOWLEDGEMENTS I would like to thank Gina Kornblith for helping with the translation of this paper. 7 CITATION INDEX 1. 2. 3. WILSON L. Biting and screaming in companion parrots. Proc Assoc Avian Vet, Portland 2000, 71 - 76 HOOIMEIJER J. Behavioral problems of cockatoos in captivity. Proc Assoc Avian Vet, New Orleans 2004, 271 - 281. HOOIMEIJER J. A Practical behavior protocol for dealing with parrots. Proc Assoc Avian Vet, Pittsburgh 2003: 177 - 181 AUTHORS ADDRESS Jan Hooimeijer, DVM CPBC Clinic for Birds, Galgenkampsweg 4, 7942 HD Meppel, The Netherlands Email:
[email protected] 362 Concurrent Master Classes Sessions . The Netherlands THE AVIAN RESPIRATORY SYSTEM : NORMAL AND PATHOLOGICAL ASPECTS P. DVM. These run mainly parallel. These included the pathogenesis and aspect of the lesions. especially during clinical examination.Respiratory system . Prof. Dipl ECVP. diagnostic techniques and anaesthesia. The inner surface of the parabronchi is covered with a network of muscle bundles. These must be taken into consideration by the practitioner. The main bronchus enters the lungs and opens caudally into the posterior thoracic and the abdominal air sac. but very efficient. Em. ABSTRACT An extensice survey was conducted of pathological findings in the lungs and air sacs of birds. the dorsal bronchi branch. The dorsobronchi split into the parabronchi. Macroscopic and microscopic designations of types of lesions are given. The avian respiratory system is the most sophisticated structure in the Animal Kingdom. In contrast to the general belief. PhD. From the main bronchus. the ventral bronchi. the avian lung is elastic. Both parts are fixed. and more caudally. the surface available for gas exchange is 10 times larger. The end terminals of this network are the openings to the atria. The total volume of the avian lungs is 50 % smaller than in mammals. University of Utrecht. The atria continue as infundibuli.Department of Veterinary Pathology. After fixation the second lung is removed and the difference in volume of the two lungs is measured (personal observations). the trachea winds subcutaneously over the breast muscles in a species-specific way. Zwart. The trachea is provided with rings of cartilage. which cross the respiratory tissue.Air sac . This is clearly demonstrated in the following setup: at post mortem only one lung is removed. especially in cranes. KEYWORDS Bird – Lung . In some avian species. Yet. The lungs are relatively small.Pathology 1 INTRODUCTION The respiratory organs of the bird reveal specific anatomical and physiological peculiarities. while the other lung remains in its position in the thorax. to reach the outer boundary of the “respiratory unit” 365 . lesions around the nares may be the result of rank-order fights. It is then collected in larger veins. the fibres of connective tissue and abnormalities can be recognized. Knemidocoptes pilae infection is a frequent aetiology of lesions around the nares. arassaris and birds of prey). The air capillaries anastomose with each other and are surrounded by blood capillaries. rich in blood vessels. However this problem is well known and is only mentioned here. Between the pneumocytes type I. 1986). 366 . There is an arterial blood supply. Blood supply to the lungs is via the arteria pulmonalis. chicks. are situated between the parabronchi. however. vagus. The basal membrane is covered on one side by the pneumocytes and on the other side by the capillary endothelium. GYLSTORFF and GRIMM (1998) recognised obliteration of the nares caused by hypertrophy of the cere as well as foreign or regurgitated material and necrotic and/or ulcerative processes. blood vessels enter into the respiratory tissue. The infundibuli. These may result in ulcerative or granulomatous lesions. the surface of which is mainly provided by microvilli. rarely lead to clinical signs such as sneezing. there are scattered. They may be the result of picking at food outside the cage. accumulation of fluid or respiratory distress. Especially in polarised light. starts near the inner surface of the parabronchial lumen. running between the parabronchi. which. Starting in these bands. Small ganglia of nerve cells are scattered throughout the lung. as well as the air capillaries. Trauma can cause marked lesions. a slide can be placed under an airsac wall and cut loose at the edges. the fibres of which run perpendicular to each other. broad bands of connective tissue. Vitamin A deficiency may lead to a variety of secondary disorders of the respiratory system. In these birds. The venous conveyance via primordial veins. small areas of taller cells provided with ciliae. are covered by very thin pneumocytes. In the avian lung. the granulated type II pneumocytes can be found. the pneumocytes are invisible as they only show up in electron microscopy. For further examination of air sacs at post mortem. delicate arterioles enter into the respiratory tissue to branch into the blood capillaries. pox can be recognised. a capillary network and venous transport. The wall is covered with a very thin epithelium. Near the entrances of the bronchi. The innervation of the air sac wall is via the rami pulmonales of the N. In young pigeons. A remarkable anatomy is seen in king penguins (Aptenodytes patagonica). At light microscopy. In canary birds. surfactant is produced (SMITH et al. which branches between the parabronchi. The air sacs consist of two thin layers of connective tissue. Pathology of the upper respiratory tract There are only a few. more extensive treatises on the pathology of the upper respiratory tract. A drop of physiological saline is added and the sample is covered with a cover glass. Short. Lymph vessels surround the arteries. In pigeons and agapornids. fights (pheasants.(SMITH et al. The delicate air capillaries branch from the infundibuli. 1986). 2) Pieces of peanuts results in local accumulation of fat (BRUNNER and MEINL 1976). 1) 2) 3) Deposition in lungs 3) Soot (anthracosis) (originating from burned coal. Yolk of ruptured egg follicle: severe catarrhal pneumonia. inducing foreign body giant cell reaction. Syngamus sp. Stomach contents: Inflammation. birefringent material in atrial walls. 5) Oxalate-Crystals produced during A. earth. bacteria.): Accumulation of black. oil. canaries). secondary lymphoid tissue may develop. diatoms etc): Granulomas with greyish. In cases of generalised amyloidosis. 6) Iron pigment (pulmonary haemosiderosis): in case of haemolytic anaemia (YAMATO et al. The character of the inflammation may be serous. purulent or chronic proliferative changes. 1) Food particles [starch granules]: (forced feeding) Distributed throughout the lungs. 1998). non-birefringent material in atrial walls. parasites and irritating gases are known aetiologies. after inhaling a formalin-based disinfectant. catarrhal. niger infection in vivo (WOBESER and SAUNDERS 1975): causes a haemorrhagic – necrotic pneumonia. Lungs Retrograde aspiration 367 . Viruses. 1) A seed causes local inflammation. The animals suffocated.Aspiration Trachea: Main Bronchus: A seed (blocks the trachea). Acute death. foreign body reaction.. Occasionally. 3) Eggs of Syngamus trachea: severe local inflammation. Mycoplasms. Trematodes (Tracheophilum sisowi in swans). 2) Pieces of plants etc: Inflammation evt. wood etc. Infectious aetiologies are: mites (esp. 7) Amyloid in muscle-bundles of atrial walls (NAKAMURA et al. We observed a severe proliferative reaction of the tracheal epithelium in Gouldian finches (Chloebia gouldiae). 1996). 8) In man: Lung carcinoma related to inhalation of dust from feathers? The trachea may be subject to a number of inflammation types. 4) Dust (pneumoconiosis) (from sand. Blood (effect of trauma). 2. 4. Proliferation of the epithelium of the main bronchi may occur in canary pox infections (presence of an epidermal growth factor). The lungs are markedly hyperaemic and serous fluid is extravasated into the respiratory capillaries and the parabronchi. keeping a bird for some period in the closed hand).The effects of Vitamin A deficiency on the trachea may be different in different birds. can cause considerable damage. elevating spots. Especially in Galliformes. In the syrinx. Heat shock (high temperatures. In case of lesions caused by claws of a cat. arrows etc. Bullets. small pieces of these tissues are present (BORST and ZWART 1976). Psittaciformes and Accipitriformes. which can be accompanied by sounds. waiting in their basket to be vaccinated. Tracheoscopy may loosen the mass. The respiratory tissue Microscopically. Mechanical lesions of the lungs Perforation of the thoracic wall may occur. The main bronchi Chronic infections or irritations may lead to the production of Bronchiolar Associated Lymphoid Tissue (BALT). 3. cornifying epithelium leading to infections. there is a danger of Pasteurella multocida infection. Generalised lesions These are: 1. Bleedings in the trachea. Inflammatory changes are seen in case of cryptosporidiosis. Suffocation of the patient can be a consequence. In Psittaciformes. eventually followed by aspiration of blood may be consequences of trauma such as collision with cables and others. metaplasia of epithelium. cartilage and bone tissue can be found as a normal constituent of the avian lung. cornified epithelial cells accumulate in the mucous glands and lead to the production of pinhead-sized whitish. may lead to a most severe respiratory distress. Inhalation of Teflon fumes. Typical example are pigeons. Inhalation of water (drowning). the mucous glands are lost and the tracheal mucosa changes into a uniform layer of stratified. In Galliformes. Aplasia Aplasia of part of a lung is a rare finding. which can then be inhaled. 368 . The subcutaneously situated venous plexus at the ventral side of the neck is overfilled. Aspiration of the material can eventually alleviate the acute danger. due to differences in anatomy and physiology. various types of lesions can be recognised 1. Technically perfect survey radiographs may reveal an indication of dilatation of parabronchi. 3) bacterial thrombi). 1996). at least part of the parabronchi dilates to ensure such a passage. compression by tumours. in principle. pulmonary tuberculosis and/or mycosis etc. is based on comparative pathology. the caudal parts of the lungs are most frequent affected. After 2-4 days of respiratory distress. Fat emboli (in 22 out of 49 psittaciformes (BRUNNER and MEINL 1976). In a minah bird (Gracula religiosa). However. 369 . and increase in connective tissue. For instance. this cannot be applied throughout. 3. 4. the first signs of disturbance of respiratory capillaries occur. This may occur in case of degeneration of the heart muscle. suffering from hydrops ascites. Changes in blood vessels 1. 8. Thromboembolic lesions (in case of bacterial inflammation such as bumble foot or endocarditis (GREENWOOD et al. With time.5. 2) diffuse intravascular coagulation. At post-mortem it should be paid attention whether the parabronchi are filled with air or with fluid or exudates. resulting in a wide parabronchial lumina. The classification given in this paper. Focal haemorrhagic-catarrhal pneumonias can be the result of viral (canary pox). Causes of emphysema are sub acute of chronic respiratory impairment such as chronic inflammation due to mite infection. there is a progressive loss of respiratory tissue. (in case of deep freezing there is a lot of free fluid – it is no longer fixed in cells). 5. Passive congestion of blood in lungs. 6. Emphysema. 7. the congestion of lungs coincided with the production of haem-crystals. a characteristic aspect is the parallel arrangement of the blood capillaries. Pneumonia Pneumonias are the most frequent cause of death in captive birds (IPPEN and SCHROEDER 1972). the infundibuli dilate. I have seen these in connection with foreign materials such as Silicium (pneumoconiosis) and plant material. a minimal amount of respiratory tissue. It is essential for the functioning of the lungs that a sufficient volume of air passes through the parabronchi. 2001). At macroscopic examination. Proliferation of Arteria pulmonalis branches in the lungs Calcium deposits in arteria. Cavernous pneumonias are rare. pseudotuberculosis) and fungal (aspergillosis in weakened birds) infections. bacterial (salmonellosis. In case of suffocation (narcosis in concentrated ether) there is an acute dilatation of the parabronchi. In case of pneumonia. At microscopy. Most probably this situation has been studied separately (ZANDVLIET et al. 2. 2. Thrombi (causes: 1) vascular invasion by Absidia corymbifera. which most probably is related to the retrograde flow of air through the lungs. Focal proliferation is seen near the adherence sites of lung mites.3. 2. Atrial walls are centres of reaction. Swelling and proliferation in case of an acute diffuse pneumonia. Tumours Primary lung-tumours are rare in birds (Table 1). A more diffuse proliferation is occasionally seen. coli. Fibrosis of lungs is very rare. Granulomatous pneumonias occur especially with bacterial (E. Loss of respiratory tissue is seen in case of emphysema due to prolonged respiratory distress or chronic infections of low intensity. 2. Only occasionally proliferation of the respiratory epithelium and/ or the capillary endothelium is recognised. very few cases are known in which the volume of respiratory tissue has not distinctly diminished and reveals a diffuse increase in connective tissue. 370 . 3. by fibroangioblastic tissue. though the aetiology could not be defined. They are involved in: 1. Fibrinous pneumonia with exudation of fibrin. avian pneumonias can be differentiated in: 1. bacteria or parasites (tachyzoites of Toxoplasma). The lymph vessels around the artery can also be involved in pneumonias. Calcium deposition on the walls of respiratory capillaries may incidentally occur. Exudation of macrophages (phagocytising fungal spores and other materials). most probably they died before this stage has developed. Eventually combined with a metaplasia to mucus producing cells. Organisation of exudates in parabronchial lumina. They also may contain lymphocytes and/or heterophils. for instance around inflammatory foci caused by bacteria of fungi. The exudate in the parabronchi had produced hyaline membranes. Small nests of lymphocytes may occur in atrial walls. The stage of hepatisation was not recognized in birds. Salmonella typhimurium. According to the type of reaction. In case of oedema. pseudotuberculosis) and fungal infections (A. The respiratory epithelium of air capillaries was cubic (and the epithelium of the atria revealed polyp-like proliferations). Though some fibrosis generally occurs in case of emphysema (as a result of inflammation). Y. fumigatus). Eventually in combination with phagocytising foreign body giant cells. 3. these lymph vessels may be overfilled. 4. Catarrhal pneumonia with exudation of heterophils and lymphocytes. Pathologic proliferation of muscle fibres in the tips of atrial walls occurs in cases of chronic irritation. with exudation of protein rich exudates and generally a marked hyperaemia. Pathologic changes in the respiratory tissue generally are very delicate. Impressive changes were found in a white-fronted goose (Anser albifrons) suffering from leucosis. It can also be a consequence of the presence of parasitestages in the endothelial cells (Plasmodium infection in penguins). Serous pneumonia. This can be a localised reaction. 1991 Tumours may cause compression of the lungs. hyperaemia was clearly recognised. Common name Golden eagle Red-tailed hawk Eider duck Scientific name Aquila chrysaetos Buteo jamaicensis Somateria mollissima Type of tumour Bile duct carcinoma Bile duct carcinoma lipomatosis/ fibromatosis Author Mikaelian et al. this may result in suffocation or pneumonia (check by redrawing the plunger). At microscopic examination. lymphocytes and macrophages could be recognised. Un-infected wounds generally heal without further treatment. the frequency is ± 1. In case fluid is accidentally injected into an air sac. We could recognise catarrhal and chronic reactions. This was seen in a chick (Gallus gallus forma domestica) suffering from an osteosarcoma of the ribs. It had resulted in extensive emphysema of the lungs. Inflammation of air sacs Acute catarrhal lesions were characterised by hyperaemia and production of an exudate. Chronic alterations were seen as a variable increase in connective tissue.Table 1: Primary lung tumours listed in birds. and local production of lymph follicles. Table 2: Metastasis of tumours to the lungs listed in birds. 1998 Metastasis of tumours to the lungs is known (Table 2).57 %. In dense colonies of terns (Sterna hirundo) and herring gulls (Larus argentatus). mites or filariae may be found. At endoscopy examination of the air sacs. In addition infiltration of heterophils. 1996 Daoust et al. aspergillomas. infiltration of inflammatory cells. Most of these cases are the result of trauma and removal of accumulated air results in a rapid healing. lesions in the air sacs are related to these in the lungs. Rupture of air sacs Rupture of air sacs are mainly caused by trauma or marked respiratory distress (pneumonia. The air sacs Generally. 1998 Hartup et al. Common name Cockatiel Grey parrot Salmon crested cockatoo Scientific name Nymphicus hollandicus Psittacus erithacus Cacatua moluccensis Type of tumour Bronchial tumour Fibrosarcoma Cystadenoma Author Burgmann1994 André and Delverdier 1999 Powers et al. endocarditis). Inflammation of muscle bundles stretching from the ribs over the ventral surface of the lungs was recognised 371 . 2 CITATION INDEX 1. et al. 42(1): 209 . This may lead to inflammation of the air sacs. Vet. 25(2): 387 391. 4. 5. 330 . 13.84. 42(2): 408 .543. 6. Pulmonary fibrosarcoma with hepatic metastases in a cockatiel (Nymphicus hollandicus). ANDRÉ JP and DELVERDIER M. Ber. 10: 94 . et al. 27(4): 539 . Vet Pathol 1976. MIKAELIAN I. 13 (3): 180 . et al. 12. Avian Dis 1998. air sacs and respiratory space of the budgerigar (Melopsittacus undulatus). J Zoo Wildl Med 1996. 27(3): 321 . The muscles were hyaline and yellowish. 15.334. Axillary cystadenocarcinoma in a Moluccan cockatoo (Cacatua moluccensis).139. SMITH BL.in a macaw. et al. SMITH JJ. Kleintierpraxis 1971. Erkrk. NORRMANN A. 3. Pneumoconiosis in the captive New Zealand Kiwi.186. 2nd Ed. In addition.412. Microscopic and submicroscopic anatomy of the parabronchi.214. 11. 14: 11 . Ulmer. 9. A contribution to diseases of zoo-birds (Ein Beitrag zu den Erkrankungen der Zoovögel). Multicentric intramuscular lipomatosis/fibromatosis in free-flying white-fronted and Canada geese. J Wildl Dis1991. HARTUP BK.27. Avian Pathol 1998. 8 (2): 81 . In case of rupture of the proventriculus (psittacine proventricular dilatation syndrome).GREENWOOD AG. 372 . Pathol 1973. 8. Fat emboli in the blood vessels of the avian lung (Fettembolische Verschlüsse in den Blutgefässen der Vogellunge). 177(2): 221 . Primary bronchial carcinoma with osseous metastasis in an African grey parrot (Psittacus erithacus). 2. Avian Pathol 1996.101. Systemic amyloidosis in laying Japanese quail. Zootiere 1972. NAKAMURA K. Avian diseases (Vogelkrankheiten). suffering from a bacterial pneumonia.179. BRUNNER P and MEINL M. food particles and even grit may be propelled into the air sacs. Bone structures in avian and mammalian lungs. J Avian Med Surg 1999. perforation of the gizzard (sharp objects).103. J Assoc Avian Vet 1994. 13: 17. BORST GHA and ZWART P. DAOUST P-Y et al. GYLSTORFF I and GRIMM F. Stuttgart 1998. The diseases of the respiratory organs in the budgerigar (Die Krankheiten der Atmungsorgane beim Wellensittich). BURGMANN PM. POWERS LV. Cholangiocarcinoma in a red-tailed hawk (Buteo jamaicensis). et al. 27(1): 135 .325. Avian Dis 1998. 13: 98 . the same muscles were invaded by sarcosporids. showing up as thickening and (brownish) discolouration. 16: 169 . 14. 7. et al. 10. Am J Anat 1986. Verhandl. et al. Metastatic cholangiocellular carcinoma and renal adenocarcinoma in a golden eagle (Aquila chrysaetos). Vet Path 1976. Vegetative endocarditis in a Waldrapp ibis (Vegetative endocarditis in a Waldrapp ibis).242. IPPEN R and SCHROEDER H-D. 1248. Avian Pathol 2001. et al. Zootiere Eskilstuna 1990. 32(2): 381 . The Netherlands. JR. Email: bibliozoo. 20.142. Avian Dis 1975. 18. 32 Int. et al. d. Weijerstraat 16. ZANDVLIET MMJM. Verhandl.1242 and 1243 . WOBESER G. Symp. J Wildl Dis 1996. WISSER J. AUTHORS ADDRRESS Peernel Zwart
[email protected]. 19: 388 . V. et al. 17. ZWART P. 43(7): 1238 . Chronic pulmonary interstitial fibrosis in Amazon parrots.384. Am J Vet Res 1982. Dept Veterinary Pathology. Hemolytic anemia in wild seaducks caused by marine oil pollution..nl 373 . Pulmonary oxalosis in association with Aspergillus niger infection in a great horned owl (Bubo virginianus). et al. Emerit Professor. WELLS R..524.392. 137 . University of Utrecht. 19. Ber. Erkrank. 30: 517 . YAMATO O. and SAUNDERS. Burg. Acute toxicosis of budgerigars (Melopsittacus undulatus) caused by pyrolysis products from heated polytetrafluoroethylene: clinical study.16. PhD. 3981 EK BUNNIK. Deposits in the air passages of zoo birds (Ablagerungen in den Luftwegen bei Vögeln aus zoologischen Gärten). gastrointestinal disease. FLUID THERAPY AND CPCR FOR THE AVIAN PATIENT Marla Lichtenberger.g.g.. surgical complications. physiological diversity and lack of research and clinical data on their response to therapy. The message of clinical importance is that fixed fluid regimens (e.. fixed volumes (e. The message of clinical importance is that fixed fluid regimens (e. can produce astounding and at times miraculous results in avian medicine. fixed volumes (e.g. United States of America SHOCK. the same principles and techniques of monitoring used in domestic animals can be applied to the exotic patient. the same principles and techniques of monitoring used in domestic animals can be applied to the avian patient.. Many of these birds present in a state of hypovolemic shock due to trauma. Treatment of hypovolemic shock and critical care monitoring in birds are complicated by small patient size. DVM. monitoring techniques. and owners want to receive high quality medical care for these pets. The goal of this article is to provide an in-depth presentation on the principles and pathophysiology of shock.The Animal Emergency Center Glendale. The goal of this presentation is to provide an in-depth discussion on the principles and pathophysiology of shock. WI. Despite these impediments. types of fluids. ml/kg) and rules of thumb are in most instances outdated. and shock resuscitation methods for use in birds. 1 INTRODUCTION Increasing numbers of birds are being kept as pets. 374 .g. monitoring techniques. types of fluids.. Arterial blood pressure measurement is an important tool in the management of the critically ill bird.. Treatment of hypovolemic shock and critical care monitoring in birds are complicated by small patient size. Despite these impediments. Principles of cardiopulmonary-cerebral resuscitation will also be discussed. Lactated Ringers). and shock resuscitation methods for use in birds. An understanding about fluid therapy and blood pressure monitoring will be discussed. physiological diversity and lack of research and clinical data on their response to therapy. DACVECC ABSTRACT Increasing number of birds are being kept as pets and owners want to receive high quality medical care for these pets. Appropriate fluid therapy. Lactated Ringers). combined with frequent patient evaluation and periodic blood pressure monitoring techniques. coagulophaties and severe live disease. inappropriate and often times inadequate. ml/kg) and rules of thumb are in most instances outdated. Exam Initially the bird should be evaluated in its cage. Often. interest in the environment. cat carrier). Proficiency in handling birds is an important factor in gaining client confidence. accurate treatment is vital to a favourable outcome. In virtually all cases. birds with chronic disease present as an emergency because of their ability to mask clinical signs of the disease until the condition is severe. Most clients have never heard their bird scream or seen their bird struggle the way it does during restraint. Following stress. seizuring. It may be better to postpone a complete physical exam in a weak bird or bird in respiratory distress. The client should be instructed to bring the bird in a cage. can produce astounding and at times miraculous results. The optimum temperatures for ill birds are 85 to 90ºF (29-30ºC). amount or blood). Once the bird has arrived. urates (normally is white to off white). Prompt. Perform a complete physical exam as the condition of the bird allows. should be picked up in a darkened room. then the bird is probably very sick and needs to be seen. if possible. The nurse should assess the condition of the bird in a room. Efficiency is important when handling a bird. combined with frequent patient evaluation and periodic blood pressure monitoring techniques. If the owner is concerned enough to call. In cases of bleeding. Prior to picking up the bird. The head should be examined for 375 . closed window and no fans. The water dish should be emptied but the cage should not be cleaned prior to travelling to the hospital. and fluffing of the feathers are assessed. Setting up ahead of time for procedures to be performed will reduce the total restraint time. weak or sitting at the bottom of the cage. Its posture. bleeding.g. ability to ambulate and perch. respiratory distress. regurgitation and anorexia are considered true emergencies. If the bird is fluffed. While all signs reported by the client can be of concern.inappropriate and often times inadequate. It is important to warn clients prior to picking up the bird what is going to happen. particularly finches and canaries. the bird should be evaluated immediately by the veterinarian. respiratory status. Examine the faeces (colour. Weak birds and birds in respiratory distress could die during handling. Most birds do not show signs of illness in the early stages of disease. birds frequently demonstrate polyuria. it is ideal to have a trained receptionist call for an immediate triage by a nurse. A quick one minute exam can be performed on a bird while taking the bird out of the cage and placing in the incubator. you should determine if it is safe to restrain the bird. The cage can be examined for discharges and vomit and sources of lead and zinc. sitting at the bottom of the cage. and urine (an increase or change in colour is abnormal) parts of the droppings. I advise the receptionist to recommend the bird be brought in for an exam. otherwise instructs the owner about suitable alternatives (e. Some birds. it is best to work in a small room with low ceilings. box. head trauma and respiratory distress. Appropriate fluid therapy. Sudden death is no uncommon with restraint of obese budgies fed an inadequate diet. they should be placed in a warmed incubator and oxygen is administered. When handling a bird. Fluid resuscitation of the patient in hypovolemic shock can be a challenge and the clinician should understand the basic pathophysiology of shock. In normal birds. The beak is examined for bleeding.. psittacosis). Normal veins refill in 1-2 seconds after depression. symmetry or fractures. have knowledge of the different types of fluid and blood pressure monitoring techniques. These drugs have been extensively investigated in the shock syndrome. Appropriate fluid therapy. Hydration is assessed by eyelid mobility. Recent studies done by the author on shock in birds. The grasp reflex of the feet will help determine weakness. it is difficult to palpate abdominal organs. has provided in-depth knowledge of a birds response to hypovolemic shock. blood pressure monitoring and characteristics of fluids. These principles will then be applied to treatment of hypovolemic shock in the avian species. increased risk of infection (i. The first part of this paper will discuss general principles of hypovolemic shock. hyperglycemia and gastric ulceration may outweigh their benefits. aspergillosis. The heart and lungs are ausculted. 376 . combined with frequent patient evaluation and blood pressure monitoring can produce favourable outcomes. The bird is also weighed on a gram scale. Their use in shock caused by haemorrhage and hypovolemia is not currently recommended. 2 SHOCK IN THE AVIAN PATIENT Shock is defined as poor tissue perfusion from either low blood flow or unevenly distributed flow. The pectoral muscle should be evaluated to judge the bird’s body condition. principles of perfusion. An increase in the distance between the caudal end of the sternum and the pubis can suggest abdominal organomegaly. The side effects of immunosuppression.oculonasal discharges and swellings. The abdomen is palpated for signs of masses or fluid distension. Glucocorticoids The use of glucocorticoids in the treatment of shock is controversial. The importance of pain management and nutrition will also be discussed. The crop is palpated for presence of food or foreign bodies. skin turgor and dry mucous membranes. The respiratory rate and effort is evaluated. redness or swelling. The oropharynx and choanal is examined for colour mucous and presence of blunted papillae.e. neoplasia or ascites. Observation of refill time of the basilic (wing) vein will estimate perfusion status. This results in an inadequate delivery of oxygen to the tissues. The second part of this paper will discuss principles of CPCR (Cardiopulmonary-Cerebral Resuscitation) and ARD (acute respiratory distress). This definition applies to all species of animals. The vent should be examined for matting. When an animal begins haemorrhaging. from their release causing an increase in extracellular fluid volume and an increase in blood pressure. LRS) are considered an important means of increasing the alkalinity of the extracellular fluid. This sends a neural signal to the vasomotor centre in the medulla oblongata. 377 . which results in inhibition of vagal parasympathetic centre and stimulation of the sympathetic centre. Potential aetiologies of absolute hypovolemia would be any cause of haemorrhage. Acute blood loss of 30-40 % of blood volume has been shown to result in 50% mortality (LD50) in mammals. surgical mishaps or a ruptured neoplasia. blood pressure decreases below a mean arterial pressure of 60 mm Hg or a systolic pressure of less than 90 mm Hg. Since this is rarely possible in the avian patient. gastrointestinal bleeding. Crystalloid fluids containing lactate. Correction of acidemia initially begins with correction of the patient’s perfusion and hydration status through the use of fluid therapy. Normasol R. The LD50 for acute blood loss in ducks has been shown in a recent study to be 60% of the total blood volume. uterus or gastrointestinal tract.e. There is a strong vasoconstriction and water retention. use of sodium bicarbonate in shock is not recommended. acetate. and gluconate (i. Plasma-Lyte. This causes a decrease in return to the left side of the heart and therefore a decrease in cardiac output. The most common cause of hypovolemic shock is haemorrhage. and antidiuretic hormone. Examples of relative hypovolemia would include severe dehydration from gastrointestinal loss. or loss into a third-body space such as the coelomic cavity. Blood gas parameters must be evaluated before considering the administration of sodium bicarbonate. including trauma. there is a decrease in blood volume and decrease in venous return to the right side of the heart. With a substantial hypovolemia. Blood loss is better tolerated in birds than in mammals. or extensive loss of plasma as in a burn patient.. Renin causes release of Angiotensin 11. The humoral response is an increase in adrenal circulating catecholamines which in turn stimulates renin release via adrenergic receptors on cells of the juxataglomerular apparatus (specialized smooth muscle cells in the afferent arterioles). coagulopathy. This causes vasoconstriction of the veins and arterioles throughout the peripheral circulatory system and increases heart rate and strength of heart contraction. 3 HYPOVOLEMIC SHOCK-PATHOPHYSIOLOGY Hypovolemic shock is caused by either an absolute or relative hypovolemia.Sodium bicarbonate The most important method of correction of severe metabolic acidosis is aimed at increasing the pH through increasing the extracellular fluid pH. aldosterone. The carotid and aortic artery baroreceptors detect a decrease in stretch due to the decrease in cardiac output. The pathophysiology of hemorrhagic shock is poorly understood in avian species. Once greater than 60% of the intravascular volume is lost. The increased heart rate and normal or increased blood pressure is the key indicator of compensatory shock. with progression of lactic acidosis. skin. A hypermetabolic state results from the humoral release of epinephrine. and normal or increased flow (bounding pulses and capillary refill less than 1 second). Acidosis leads to cellular swelling with loss of extracellular fluid volume into the cells. the neuroendocrine responses to hypovolemia become ineffective and irreversible organ failure begins. 378 . There is an uneven distribution of blood flow. which causes lactic acidosis. prolonged capillary refill time. and muscles. tachycardia. gastrointestinal tract. b. there is development of multiple organ dysfunction. decreased blood pressure. Aggressive fluid therapy to support blood pressure and heart rate is required in this stage. This is the stage seen commonly in birds with blood loss less than 20% of their total body weight. Clinical signs in the bird include an increase in heart rate. Anaerobic metabolism is less efficient than aerobic metabolism. cool limbs and skin. Volume replacement at this stage is usually associated with a good outcome. That study may show that birds have a baroreceptor response to shock similar to that seen in mammals. 4 HYPOVOLEMIC SHOCK-COMPENSATORY RESPONSE a. There is a reduction in the blood flow to the kidneys. This stage occurs when fluid losses continue.A recent hemorrhagic shock study in mallard ducks (Anas platyrhynchos) documented an increase in heart rate and decrease in blood pressure following acute blood loss. Oxygen consumption in the tissues becomes dependent on oxygen delivery and anaerobic metabolism. normal or increased blood pressure. c. Early decompensatory The second stage of shock is called the middle or early decompensatory stage of shock. Clinical signs of early decompensatory shock in the bird include hypothermia. and mental depression. In anaerobic metabolism there is production of lactic acid and hydrogen ions. Irreversible hypovolemic shock occurs when the neuroendocrine compensatory responses fail to restore and maintain tissue perfusion. As cell integrity is lost. Blood pressure increases due to the increase in cardiac output and systemic vascular resistance. Decompensatory or terminal shock The final stage is called decompensatory or terminal shock in all animals. pale mucous membranes. which lead to metabolic acidosis after exhaustion of the buffer systems. Early or compensatory The early or compensated stage of shock occurs as a result of the baroreceptormediated release of catecholamines. For animals with evidence of interstitial dehydration on physical examination but stable cardiovascular parameters. weak or absent pulses. severe hypotension. and anticipated ongoing losses. If the interstitial volume is rapidly lost. pulmonary ooedema. These parameters change in response to the various stages of shock as was discussed above. hypothermia. 5 PERFUSION VERSUS HYDRATION The transport of fluid and oxygen through the blood vessels to the capillaries is called perfusion. Interstitial hydration is the presence of fluid in the interstitial space. Interstitial volume deficits are typically associated with a decrease in skin turgor and dry mucous membranes. The rate of fluid administration depends primarily on the rate of fluid losses and clinical status of the animal. so this method is usually not warranted. pale or cyanotic mucous membranes. then the interstitial fluid deficit should be rapidly replaced (2-4 hours). capillary refill time.At this stage. 6 CLINICAL TECHNIQUES FOR THE MEASUREMENT OF BLOOD PRESSURE The “gold standard” for measuring blood pressure would be the direct method. and the heart and brain begin to fail with the severe hypoxia. Rehydration of the interstitial compartment is best accomplished using an isotonic replacement fluid. Isotonic replacement fluids are administered according to the patient’s estimated dehydration. oliguria to anuric renal failure. Tissue perfusion is directly dependent on adequate intravascular volume and a normally functioning cardiovascular system. Cardiopulmonary arrest commonly occurs. except that the damage has overwhelmed the body’s natural protective mechanisms and multiple organ failure has occurred. and a stupor to comatose state. Unfortunately. Clinical parameters evaluated during assessment of interstitial hydration include mucous membrane moisture. which include mucous membrane colour. Perfusion should be evaluated first followed by an evaluation of hydration. as indicated by the physical exam and laboratory parameters. The pathophysiological profile of terminal shock is a continuum of that described for the early decompensatory stage. position of the ocular globe. fluid deficits can be replaced over 12-24 hours. The effects of perfusion are demonstrated on physical examination by perfusion parameters. Adequate hydration provides support for the cells and transport media for molecules. the intrinsic compensatory mechanisms no longer provide delivery of oxygen to the tissues. arterial catheterization in birds is technically difficult and can be time-consuming. Although there are several indirect 379 . absent capillary refill time. maintenance needs. When dehydration is estimated to be greater than 8%. and skin turgor. an intravascular volume deficit is expected to occur from osmotic fluid shifts. and resting heart rate as well as blood pressure. The clinical signs are bradycardia with low cardiac output. which entails placing a catheter in an artery and connecting it to a pressure transducer. Reticulocyte release from the bone marrow in mammals occurs rarely after 2-4 days and most commonly longer than 5 days after acute blood loss. The cuff is deflated with the first sound heard and marked as the systolic pressure. 7 RED BLOOD CELL REGENERATION AFTER ACUTE BLOOD LOSS After acute blood loss. Crystalloids are the mainstay of the rehydration and maintenance phases of fluid therapy. Aloha.. allowing classification of anaemias into regenerative or non-regenerative types. The cuff bladder is inflated to a suprasystemic pressure with cut-off of the Doppler signal. OR) uses ultrasonic waves to detect and make audible blood flow in an artery distal to the blood pressure cuff.. A previous study done by the author. The Doppler probe is placed on the basilica and metatarsal arteries respectively. In response to tissue hypoxia. is the hallmark of intensified erythropoiesis in mammals and birds. The three basic groups (i.or non-invasive methods (i.e. mammals are dependent on red blood cell (RBC) regeneration to maintain oxygen delivery to the tissues. oscillometric and Doppler) available. synthetic colloids. Normal systolic blood pressure for birds under isoflurane anaesthesia. the cuff is placed on the distal humerus or femur. erythropoietin stimulates RBC production by the bone marrow. crystalloids. The relatively short live span of the red blood cell (28-45 days) and presence of a nucleated red blood cell (RBC) may account for a birds’ ability to mount a very early regenerative response. and Haemoglobin-Based Oxygen Carriers) of fluids will be discussed.e. it is sometimes impossible to obtain a reading on the avian patient. The use of early supportive care with fluid therapy in avian shock may help bridge the gap for the first 24 hours. Crystalloids: Crystalloids (also called replacement fluids) are fluids containing sodium chloride and other solutes that are capable of distributing to all body fluid compartments. on acute blood loss in the duck documented a early regenerative response shown by presence of polychromasia starting at 12 hours after blood loss. Crystalloids solutions can be used together with colloids during the resuscitation phase. In the birds. The Doppler method is more versatile than the oscillometric method.. Trends of change in systolic pressures obtained using the Doppler flow detector has been found to correlate well with direct pressure measurements in anesthetized ducks (unpublished data done by the author). after which birds can mount their own RBC regenerative response. or polychromasia. Reticulocytosis. type. Replacement fluids have electrolyte concentrations that resemble extracellular fluid 380 . 8 FLUID SELECTION Individual characteristics of available fluids influence the dose. and volume of fluid administered. a. measured at the author’s clinic is between 90-150 mmHg. and is the method used by the author for all exotic patients. The ultrasonic Doppler flow detector (Parks Medical Electronics Inc. Deerfield. producing a final intravascular volume greater than the volume infused. Hydroxyethyl starch is a molecule made from maize or sorghum and is primarily an amylopectin. Colloids Colloids are fluids containing large molecular weight substances that generally are not able to pass through capillary membranes. crystalloids should be thought of as interstitial dehydrators. hypercalcemia. Interstitial oedema may be extremely detrimental in cases of cerebral oedema and pulmonary oedema. ILL). Normal saline can also be used for treating conditions causing hyperkalemia (e. Their negative charge attracts sodium and water (Gibbs-Donnan effect). Overland Park. Since most patients in shock require sustained intravascular volume expansion. Ill) and Normosol-R (CEVA Laboratories. b. Colloids can be considered intravascular volume expanders. 381 . and albumin. the highest sodium-containing isotonic crystalloid. to animals with head trauma. Thus. Examples include synthetic colloids such as hetastarch (Braun Medical Inc. not intravascular volume expanders. KS) or Plasmalyte-A (Baxter.g.whereas maintenance fluids contain much less sodium (40-60 mEq/l) and more potassium (15-30 mEq/l). The most commonly used replacement fluids are 0.9% sodium chloride. Most hospitals do not have readily available donors for species specific donation and commercial blood banks do not carry exotic pet blood products. Since approximately 80% of extracellular fluid is in the interstitial space. If the patient has lost clotting factors. North Chicago. ILL). The liver must metabolize lactate whereas many cells in the body metabolize acetate and gluconate. urinary obstruction and oliguric renal failure). It expands the volume by about 1. plasma. colloids are indicated frequently during fluid resuscitation. Buffered solutions are usually indicated for resuscitating patients in shock since administration of a highly acidic solution may worsen a preexisting metabolic acidosis. and biologic colloids such as whole blood. Therefore fluids containing lactate may not be tolerated well by animals with end-stage liver disease. Irvine..4 times the volume infused. Ideally blood should be used for resuscitation if the patient has lost whole blood. crystalloids will rapidly redistribute and after approximately 1 hour there will be only 20% of the administered volume remaining in the circulation. There is evidence to support the use of 0. This increase in interstitial fluid can lead to tissue oedema (thus decreasing the ability of oxygen to diffuse to the cells). and hypochloremic metabolic alkalosis. The lower the haematocrit becomes. lactated Ringer’s solution (Abbott lab. Deerfield. the more important this is to ensure adequate oxygen delivery to the cells. This synthetic colloid fluid contains large-molecular-weight particles that effectively increase the COP beyond what can be obtained with blood product infusion alone. They maintain intravascular osmotic pressure because their molecular size is too large to pass through the normal capillary pores.9% saline (Baxter. to avoid rapid changes in osmolality. The availability of blood products in sufficient quantities to meet the needs of the avian species is often the limiting factor in survival. CA). Buffered solutions usually contain lactate. clotting factors should be replaced. but this effect will be short-lived. gluconate or acetate. On a short-term basis crystalloids will expand the intravascular space. Although HBOC are colloids. Haemoglobin-Based Oxygen Carriers Haemoglobin-based oxygen carriers (HBOC) are indicated during resuscitation when increased oxygen delivery to tissues is desired. they have the added advantage of carrying oxygen to the tissues. 3-DPG levels leading to increased oxygen binding and decreased oxygen delivery at the tissue level. Oxyglobin® can also be administered to patients suffering from chronic anaemia with PCV values less than 10%. Because it is a smaller molecule than red cells it is able to perfuse tissue beds that red cells may not be able to reach. Filters are not required. the length of clinical benefit is currently unknown Oxyglobin® can be administered at any time to any species in need of a blood transfusion or during acute and critical situations in which an acute increase in blood volume and oxygen carrying capacity are needed. It has a pH of 7. It has a short half-life (30 to 40 hours). Its oxygen affinity is dependent upon the chloride ion concentration not the concentration of 2. similar to that of plasma. low volumes of Oxyglobin® can be used effectively to treat hemorrhagic shock. It has a COP (20-25 mmHg). 382 . 3-DPG). Once opened the bag must be discarded within 24 hours due to the production of methaemoglobin. Oxyglobin® is now available in 60 ml containers. It is isoosmotic and has an average molecular weight of 200. For this reason. which makes it useful for hospitals that cannot keep blood products readily available.8 which means it will not contribute to the acidosis of the patient during infusion. It is an excellent fluid to use in animals suffering form acute blood loss secondary to trauma or surgical mishap and should be administered to patients that have become haemodiluted (PCV< 15-18%) from crystalloids. Synthetic colloids are administered with isotonic crystalloids to reduce interstitial volume depletion. Oxyglobin® is a purified. c. as many crystalloid solutions may.Hetastarch has an average molecular weight of 450. It has a lower viscosity than canine blood that may improve microvascular flow.000 Daltons and has a half-life of 25 hours. It can be kept at room temperature and has a 3year shelf life. polymerized bovine haemoglobin that is in a modified lactated Ringer’s solution that is approved for use in dogs. This provides a distinct advantage over blood that has been stored longer than 1 week that may have significantly depleted 2.000 making it a very effective colloid. 3-diphosphoglycerate (2. Oxyglobin® can be administered via standard intravenous administration sets and standard intravenous infusion pumps can be used for delivery. however. Oxyglobin® is up to 10 times more effective than blood when given during fluid resuscitation to animals in hemorrhagic shock. it offers the opportunity to decrease patient morbidity and mortality. Because it contains no antigens cross matching is not required and there is no possibility of transfusion reactions. In addition the normal oxygen affinity of Oxyglobin® is lower than that of normal canine blood which enhances delivery of oxygen to the tissues. Appropriately used. The dose of crystalloid administered is only 40 to 60% of what it would be if crystalloids were used alone during resuscitation. ILL). and restore intracellular water balance. in ducks. Resuscitation implies an urgent need to restore tissue perfusion and oxygenation. should immediately be placed in a warm incubator (Temperature at 85-90°F [29. North Chicago. the importance of small volume resuscitation with one bolus of crystalloids with Oxyglobin® to hypovolemic birds may have clinical relevance. A pilot study of acute hemorrhagic shock in cockatiels treated with HES and Oxyglobin® found no adverse side effects associated with either fluid. 10 FLUID THERAPY-BIRDS Any sick. The maintenance phase provides fluids and electrolytes to replace ongoing losses. The intravascular volume must be replaced initially. However. Recommendations for fluid resuscitation subsequent to severe blood loss in birds have included the administration of whole blood.9 FLUID THERAPY PLAN The administration of fluids is almost always thought of as the single most important therapy during surgery and for the treatment of patients suffering from hypovolemic shock. this must be stopped immediately.1°C]) with oxygen supplementation for 2-4 hours. The type. Birds should be offered food and water during this time.4-32. bolus infusion of Oxyglobin® and crystalloid was effective in correcting heart rate and blood pressure within 30 to 120 seconds after infusion. Re-evaluation of the hydration status is necessary before planning the rehydration phase. When active external haemorrhage is present. responsive) and can be safely anesthetised with mask isoflurane (Abbott Lab. rehydration (correction of interstitial deficits). diagnostics and treatment for hypovolemia and dehydration can be performed. The fluid therapy plan typically has a resuscitation (correction of perfusion deficits). quantity. IO or SQ. debilitated bird presenting for emergency care. Since fluid resuscitation in critically ill birds is difficult. and maintenance phase. Rapid bolus administration (doses of 5 ml/kg over 1 minute) of Oxyglobin® or HES with crystalloids (10 ml/kg) has been given to hypovolemic birds (birds with acute blood loss) in research and clinical trials without side effects. meet metabolic demands. In a study on acute blood loss (loss of 60% of their total blood volume). Blood pressure monitoring using Doppler and an ECG can be used during these procedures. the use of avian blood products is limited by the lack of availability. Most birds benefit from the administration of warmed crystalloids at 3 ml/100gBW IV. When the bird appears stable (alert. The beneficial effects of fluid administration are most obvious in haemorrhaging or traumatized patients because of the absolute losses of blood and fluids from the vascular and interstitial fluid compartments. 383 . North Chicago. and rate of fluid administration required to reach the desired resuscitation endpoints are determined and depend on the type of shock and underlying disease process. ILL) or sevoflurane (Abbott Lab. St. however. The blood pressure of various avian species under isoflurane or sevoflurane anaesthesia at the author’s clinic is 90-140 mmHg systolic. Glendora. and the bird is released. Hold the bird in an upright position with the neck extended. when perfusion parameters and kidney parameters are normal. CA) is recommended. Tube feedings should be warmed in hot water to body temperature to avoid crop stasis. over the tongue. considerable care should be taken to ensure the bird does not bite the feeding tube and swallow the end. release the bird and allow the bird to clear the food from 384 .5 mg/kg IM) can be used for pain relief. When blood pressures are below 90 mmHg systolic. maldigestion. the tube or feeding needle is carefully withdrawn. Palpate the crop before feeding. The importance of nutrition early on is so important and emphasizes the saying “when the gut works. A curved metal feeding needle with a balled tip (Dosing Needles. Administration of analgesics is strongly recommended in these birds. Daily maintenance fluid requirements (2 ml/kg/hr) are added to the fluid deficit volume. Bolus administration of crystalloids (10 ml/kg) and colloids (HES or Oxyglobin® at 5 ml/kg) can be given IV or IO until blood pressure is greater than 90mmHg systolic. Estimation of the fluid deficit is based on estimated dehydration and body weight: Estimated Dehydration (%) x Body Weight (g) = Fluid Deficit (ml). and weight loss need nutritional support.5-1 mg/ kg) intramuscularly has been shown as a useful analgesic is birds. Ejay International. MO) can be used. use it”. Red rubber feeding tubes (Sovereign Feeding Tubes. The tube or feeding needle is gently passed into the left side of the beak. Do not feed a bird with food still present in the crop. The formula is infused into the crop. Louis. Hand-feeding formulas for baby birds can also be used. If the bird refluxes into the mouth during the procedure.The Doppler cuff can be placed on the distal humerus or femur and Doppler probe on the medial surface of the proximal ulna or tibiotarsus respectively. In the authors experience 1 or 2 bolus infusions are usually required.0 to 2. and down the right side of the oesophagus into the crop. Palpate the crop and verify that the feeding needle or tube is in the crop. Butorphanol (0. Perform other procedures before tube feeding to prevent regurgitation and aspiration. There are several enteric feeding formulas on the market which range from less than 1. Sherwood Medical. The volume is estimated at 3ml per 100g of bird. birds are treated for hypovolemia as given below. Baby birds may be fed up to 5ml per 100g as they have a greater crop capacity.0 Kcal/ml. 11 PAIN MANAGEMENT IN THE CRITICAL AVIAN PATIENT Most seriously ill birds experience pain and anxiety during hospitalization. In severely dehydrated birds that are not eating IV or IO catheters are placed for replacement of dehydration losses with crystalloids. Non-steroidal antiinflammatory drugs as meloxicam (0. 12 NUTRITION Critically-ill avian patients suffering from anorexia. The goal is to use these monitoring devices to identify cardiovascular instability early.its mouth. An electrocardiogram. Epinephrine (0. because of risk for laceration of the coronary vessels. IO (intraosseous). or via the endotracheal route (using a tom cat catheter inserted down the endotracheal tube and doubling the dose used for IV). The most recent International Heart Association guidelines for CPCR and emergency cardiac care in humans were published in 2000.01 mg/kg) and atropine (0. The primary focus was use of evidence-based medicine. Place an endotracheal tube and start positive-pressure ventilation with 100% oxygen. An alternative method for positive pressure ventilation is placement of an air-sac breathing tube. Always monitor birds with a Doppler blood pressure and electrocardiogram when placing under gas anaesthesia. If the bird arrests on anaesthesia. 385 . Cardiac arrest carries a poor prognosis. oesophagus or crop injuries may need to have a feeding tube placed into the proventriculus. The suggested volumes of food to be administered in different species. Because birds do not have a diaphragm. I have taken this information and applied the same principles to the avian species. and end-tidal CO2 monitor can be used to evaluate the effectiveness of CPR resuscitation. and can be left in place until the bird starts eating on their own. The feedings should be done three to four times a day.1 ml for a small bird) IM (intramuscularly) to stimulate the respiratory centres. because direct compression of the heart is not possible because of the overlying sternum. Intracardiac injections should be avoided.04 mg/kg) can be given IV (intravenous). stop the anaesthesia administration. The prognosis for respiratory arrest especially when caused by isoflurane anaesthesia overdose is good. is given below. Persistently regurgitating birds or birds with beak.2 ml for a large bird and 0. closed-chest compressions in birds cannot utilize the thoracic pump mechanism to increase overall negative intrathoracic pressure. Suggested Volumes for Tube Feeding Volumes Species of bird (ml) Budgerigar 1-3 Lovebird 2-3 Cockatiel 3-6 Conure 4-12 Parrot 15-35 Cockatoo 20-40 Large Macaw 35-60 13 CARDIOPULMONARY-CEREBRAL RESUSCITATION CPCR is a comprehensive term used to describe both the basic principles of cardiopulmonary resuscitation (CPR) as well as advanced life support and post-resuscitation care. Doppler blood pressure. Early recognition of cardiovascular instability is the key to success in avian medicine. The bird is given an injection of naloxone (0. This tube is termed an oesophagostomy tube. The word cerebral resuscitation was added to cardiopulmonary resuscitation to identify the importance of being alive with complete neurological function. These guidelines were used to extrapolate the basic principles for mammals. egg-related peritonitis.14 ACUTE RESPIRATORY DISTRESS (ARD) Differential diagnosis a. trimethoprim sulfadiazine) and bronchodilators when aspiration pneumonia is suspected. Causes of parenchymal disease include: cardiogenic pulmonary oedema. iii. pneumonia. mass. aerosols and candles. i.g. serology antibody test and a PCR test) to lessen chances of falsepositive and false-positve results. poxvirus. and the fluoroquinolones). Oxygen supplementation by mask or cage oxygen administered during resuscitative procedures. b. smoke inhalation. b. 386 .g.g. East Hanover. Upper airway obstruction requires rapid anaesthetic induction with sevaflurane/isoflurane anaesthesia. Birds with a history of regurgitation or vomiting may benefit from broadspectrum antibiotics (e. Gram-negative bacteria. liver disease.. NJ) at 10-15 mg/kg BID for 4 weeks. Birds suspected of psittacosis (leukocytosis . tetracyclines.e. iv. chloramphenicol. Small airway disease includes inhalant respiratory irritants as smoke. The author recommends the use of two tests run simultaneously (e. Treatment Resuscitative intervention is required most often prior to aggressive diagnostics to minimize stress.. ii. Chlamydophila. An air sac tube may partially improve respiration in birds with primary lung and air sac disease. macrolides.Birds suspected of aspergillosis (leukocytosis and pulmonary nodules) should be treated with terbinafine hydrochloride (Lamisil. Norvartis Pharmaceuticals. Teflon toxicity. Careful auscultation for a heart murmur or gallop or persistent arrhythmia suggestive of cardiogenic oedema. Causes of coelom space disease include: heart disease. Pacheco’s disease and reovirus.. Causes of large airway disease include: obstruction by foreign body (i. diarrhoea and respiratory distress. organomegaly. If there is a total upper airway obstruction. hypoalbuminemia. Parenchymal disease may benefit from use of parental or nebulisation of bronchodilators and antibiotics. seed in cockatiels). d. administration of furosemide (2-4 mg/kg IV) and nitroglycerine ointment on the back is administered and the bird is allowed to stabilize prior to further diagnostics. rapid intubation and ventilation with 100% oxygen. c. cephalosporins. Aspergillus granuloma or oropharyngeal granulomas. emergency air sac tube placement is performed. There is no “gold standard” test that can be used to unequivocally determine if a particular test is accurate. Aspergillus.. If heart failure is suspected. a. clinical signs varying from chronic unthriftiness to acute anorexia. enrofloxacin. and “lime green urates”) should be treated with drugs inhibiting the growth of Chlamydophila (e. Coelomic distension will require immediate coelomocentesis when ascites is present. Small airway disease will require: Bronchodilators (terbutaline at 0. i. Radiology or ultrasonography is useful when differentiation is not obvious. iii. egg binding or a mass lesion is suspected when the coelom is firm on palpation. Coelomocentesis is done on the right lateral coelom just cranial to the cloaca. Organomegaly. WI United States of America Email: AECMARLA@AOL. Often ultrasound can help guide thoracentesis if the fluid is loculated and difficult to aspirate. iv.01mg/kg IM) can be given parenterally or terbutaline can be given by nebulisation. Use a 25-22 gauge needle and 12 ml syringe.c. Save any fluid recovered for culture/sensitivity and cytology. ii. d. AUTHORS ADDRESS The Animal Emergency Center.COM 387 . Glendale. 4%.8% of their body mass as kidney tissue. Important consideration should be given to the bird’s diet and exposure to toxins and infectious agents in addition to a variety of diagnostic tests. while those with salt glands have 1. Dipl ABVP (Avian).Water excretion – Urinalysis – Casts – Electrophoresis – Urography . and the Lafeber Company2. The urinary concentrating ability generally varies inversely with body mass. Birds without salt glands have 0. DVM. Their kidneys are retroperitoneal in location.Protein levels . Orosz 2 PhD. diagnosis and treatment of renal disease when compared with mammals. However. USA. avian renal surgery is often restricted and therapy is frequently limited to supportive care. The unique anatomic and physiologic features of avian kidneys lead to many differences in the types.Renal fibrosis – Amyloidosis – Hyperuricemia . Dipl ABVP (Avian) and S. Perrysburg. Echols1 DVM. Concentration of urine may occur by retroperistalsis. US and Perrysburg Animal Care1. The avian kidney contains 2 basic types of nephrons—cortical or reptilian-type nephrons and the medullary or mammalian-type nephrons that contain medullary loops of Henle. Ohio. Texas. 388 . 1 GROSS ANATOMICAL CONCEPTS The kidneys of birds are firmly attached to the synsacrum and sit within the renal fossae. Extensions of the abdominal air sacs from the pelvis can be found between the synsacrum and the kidneys. Cornell. Austin. Illinois. Tennessee. United States of America A MASTER CLASS OF THE URINARY SYSTEM : FROM ANATOMY TO TREATMENT OPTIONS M.Renal disease .E.Gout ABSTRACT The kidneys of birds are retroperitoneal and firmly attached to the synsacrum.Calcium levels .S. renal biopsy and histologic evaluation is often the only means of a definitive diagnosis and can better guide the clinician to more accurately treat kidney disorders in birds.Glomerular filtration – Diuresis . The University of Tennessee. Knoxville. Dipl ECAMS KEYWORDS Urinary system – Kidneys .Westgate Pet and Bird Hospital1. Given anatomical constraints. College of Veterinary Medicine2. and extend from the lungs cranially to the distal end of the synsacrum caudally. This stalk-like region of the narrowed end of the lobule is the medullary region or the medullary cone.The kidneys of birds do not have lobes as in mammals but have anatomical divisions. the basic unit of the avian kidney is the lobule. The wide. They tend to be pear-shaped and wedged between the interlobular veins of the renal portal system. and caudal division. This area contains collecting tubules and the nephronal loops of Henle from the medullary type nephrons. 2 HISTOLOGIC STRUCTURE In general. middle. 389 . Parts of a cortical (reptilian) nephron type (top) and a medullary (mammalian) nephron type (bottom). Each kidney has a cranial. but they are often obscured. Figure 1. The widest portion of the lobule sits near the surface of the kidney. From a clinical perspective. while the smaller end is where the collecting tubules converge to form branches of the ureter. Renal lobes may be recognized in some species. superficial portion represents the cortical region of the lobule and contains nephrons of both the cortical and medullary type nephrons. it is important to note that the spinal nerves from the lumbar and sacral plexuses pass through the substance of the kidney. Nerve impairment to the pelvic limb from swelling or pressure as with neoplasia can result in neuropraxis or proprioceptive deficits to the limb. The lobules are oriented with their long axis perpendicular to the long axis of the kidney as it sits in the renal fossae. an epithelial thickening at the beginning of the distal convoluted tubule. The medullary type has a loop of Henle. Birds. Amyloid deposits are often related to chronic inflammatory disease and usually found systemically. The outer ring of collecting tubules of each lobule consists of perilobular collecting tubules. 3 GENERAL MECHANISMS AND CONSEQUENCES OF RENAL INJURY Initiation of renal disease Proposed mechanisms of the process of initiation of renal injury and perpetuation of disease are complex. This is the type of nephron found in reptiles and the nephrons secrete primarily uric acid.and intralobular veins. they form a secondary branch of the ureter. The products resulting from the arachidonic acid cascade have effects throughout the body. a collecting duct. 390 . but can affect specific tissues. which is anatomically similar to the short mammalian nephron type with the proximal descending loop enlarging prior to the bend. An inner ring is formed by the ascending limbs of the loops of Henle. like mammals. Amyloidosis Amyloidosis is occasionally noted in association with avian renal disease. Commonly. the avian kidney contains 2 basic types of nephrons—cortical or reptilian-type nephrons and the medullary or mammalian-type nephrons that contain medullary loops of Henle. these medullary tubules combine to form a single larger vessel. They form the majority of the nephrons in the kidneys of birds. All of the nephrons have a proximal and distal portion to their nephrons. The cortical type of nephron is found in the cortical region only and does not have loops of Henle. have a juxtaglomerular apparatus. but the mammalian model may shed some light on avian renal disease. which consists of a macula densa. The distal segment of the reptilian nephron has compact convolutions near the intralobular veins. When several of these collecting ducts combine. However. For this reason. Both types of nephrons have a renal corpuscle at their proximal ends. The middle segment of this nephron is poorly defined but is short and convoluted.As suggested. as their intermediate segment. The renal corpuscles lie about midway between the inter. the loop of Henle. as occurs in mammals that has a potential to produce and. but only the mammalian type have the loop of Henle. The cortical nephrons have their proximal segment in the outer portion of the lobule and form approximately half the length of this nephron type. some of the inflammatory cascade that occurs with renal disease is described. a recent study suggests a gradient of nephrons with the outermost representing cortical nephrons and the innermost nephrons (with the largest loops of Henle) representing the mammalian types. while those that are part of the medullary region are termed medullary collecting tubules. The medullary nephrons have. concentrate urine. to a certain extent. Post-mortem renal change Renal post-mortem changes are noted in chickens as soon as 22 minutes following death at 37oC (98. cyclo-oxygenase upon arachidonic acid results in the formation of numerous prostaglandins (PE2. anorexia. disorientation. blood flow. tubules. In mammals. and PGD2) and thromboxanes (thromboxane A2 [TXA] and thromboxane B2) all of which have varying actions on cells. demineralization of bones. visceral gout. Selected toxic and nutritional diseases Vitamin D intoxication: As a result of excessive calcium uptake. The action of prostaglandin synthetase. Early renal post-mortem changes occur in the proximal tubular epithelium. renal medulla tubular cells. Nephritis: Nephritis is simply inflammation of the kidney and may involve the interstitium. recruitment of inflammatory cells. polydipsia. prostaglandin and thromboxane production is altered (primarily increased). polyuria. nausea. and muscle weakness. Nephrosis: Nephrosis is a non-specific histopathologic change characterised as any degenerative. and especially platelets. TXA causes mesangial cell contraction and is a potent vasoconstrictor. 391 . visceral calcinosis. nephrocalcinosis. Platelet aggregation and activation occur secondary to complement activated antigenantibody interactions and renal endothelial damage. Symptoms of hypervitaminosis D include hypercalcemia. and/or the glomerulus (although ‘glomerulonephritis’ is typically reserved for glomerular lesions). Hypervitaminosis D may occur when feeding chicks vitamin D containing supplements.The renal medulla and papilla are a rich source of an enzymatic group known as prostaglandin synthetases. and other physiologic effects.6oF). and urate nephrosis are considered frequent complications of vitamin D intoxication in birds. painful joints. These ‘alterations’ subsequently result in varying effects on the body and kidney including changes in renal vascular resistance. non-inflammatory lesion of the kidney. followed by collecting tubule epithelium and glomerular nuclei. Thromboxane A2 is produced by glomerular epithelial and mesangial cells. from cloudy swelling to necrosis. Both of these actions can result in decreased glomerular filtration rate (GFR) resulting in renal damage. PGF2�. In response to renal ischemia and vasoconstriction. Haemostatic abnormalities: Abnormalities of haemostasis can be noted. whatever the cause. Non-steroidal anti-inflammatory drugs act to inhibit prostaglandin synthetase and represent another method to ‘alter’ these arachidonic acid by-products and their subsequent actions. Thromboxane A2 also promotes platelet aggregation and may be partially responsible for haemostatic abnormalities noted with renal disease. TXA production is considered the main cause of renal vasoconstriction associated with acute renal failure and is believed to play a pathogenic role in many forms of kidney disease. Because of the multiple sources of toxin. In a study involving young and adult budgerigars (Melopsittacus undulatus). High protein diets: High protein diets have been associated with renal disease in birds. These epithelial changes can result in decreased mucin production and excessive keratin leading to plug formation and ureteral obstruction. has been reported in chickens and ducks.5% calcium. and 3000 IU/kg/feed of vitamin D3. Parent birds were fed diets containing 0. Renal lesions may include proximal tubular necrosis and degeneration (nephrosis). Dietary supplementation with calcium. 1000. 0. visceral gout. but a direct relationship with renal disease has not been established. Ochratoxin A is produced by several species of Aspergillus and Penicillium.5 % calcium died by 24-32 days old and never fledged (32-35 days). The adults subsequently fed the young the same diet.5% calcium. While only a few adults died by 5 months on diets containing 1. such as budgerigars. Lead nephropathy: Lead toxicity is the most common cause of metal poisoning in waterfowl and affects a wide variety of birds. Lesions have been limited to non-specific tubular nephrosis and were reversible after feeding a non-pelletised diet for 1-3 months.7% and 1. and in some birds. has been correlated with renal disease. and 1.3%. Diets high in urea have also been linked to nephritis outbreaks in poultry.3% calcium had no evidence of metastatic mineralization and had good hatchability and growth rates. multiple avian species can be exposed to ochratoxin resulting in kidney damage. most had metastatic renal mineralization when fed 0.7%.5% dietary calcium. Fish-meal adulterated with urea was linked to high (6-8%) mortality in 2 separate farms. When fed a diet containing 3000 IU/kg/feed of vitamin D3 there was a questionably increased mortality rate only in the birds receiving 1. increasing dietary calcium levels were shown to be more renal toxic than was excess vitamin D3.5% dietary calcium with a range of 500. Birds fed 0.7% calcium.5% calcium. acidfast intranuclear inclusion bodies. as well as limestone sand substrate. there was a clear correlation with mild and severe metastatic (renal) mineralization in birds fed 0. may be very sensitive to dietary calcium levels and that supplementation should be used cautiously. 1500. but only under specific conditions. Hypovitaminosis A: Hypovitaminosis A may also lead to renal disease in avian patients. In birds with hypovitaminosis A. However. The young birds fed 0. the ureters and renal collecting ducts may undergo metaplasia. Mycotoxic nephropathy: Mycotoxic nephropathy. ‘Diet-induced renal disease of colour variety psittacine birds’: There may be form of renal disease induced by feeding predominately pelletised diets to colour mutations in psittacine birds. This study suggests that some species. 392 . Extraordinarily high protein levels in the diet of genetically predisposed chickens have been shown to cause gout.7% and 1. respectively.Hypercalcinosis: High calcium intake has also been directly correlated with renal disease in birds. Growth rate and hatchability were poor only in the groups fed 1. Ochratoxicosis induces hypertrophy of renal proximal convoluted tubules. primarily due to ochratoxin A. the blood then flows into renal veins of its appropriate division. Depending on the division of the kidney. If 393 . This has important ramifications for the removal of urates along with other components including drugs. The renal portal system is involved in the secretion of urates in the blood so that it can be excreted by the kidneys in birds. These valves are found within the lumen of the common iliac veins between the renal and the portal veins.4 THE VENOUS SUPPLY TO THE KIDNEYS Blood from the area around the loops of Henle are drained by the venulae recta into the intralobular veins. The renal portal system forms a venous ring with both kidneys. It has been suggested that the renal portal system provides about two-thirds of the blood supply to the kidneys that bypasses the glomeruli. There are numerous small venous branches that take origin from this ring to penetrate the parenchyma of the kidney. Figure 2. Ventral view of kidneys of the domestic chicken illustrating the renal portal system. Renal portal valves control blood flow to the kidney and systematically. The intralobular veins drain blood into the efferent renal veins or branches. Each valve is innervated by adrengeric and acetylcholine receptors. Spontaneous articular gout in birds without underlying renal pathology is relatively uncommon and appears to have a hereditary basis. This shunt can result in the bypass of the kidney completely but often is only partially activated so that blood may go a variety of directions. then blood will flow directly into the caudal vena cava and not the kidney. inflammatory bowel disease (IBD) can be related to renal disorders. colitis may serve as a source of infectious agents. Therefore. administered various nephrotoxic agents. and general kidney damage can decrease uric acid elimination. In humans. even if the initial insult is treated and ‘cured’. Intestinal inflammation may lead to renal disease. into the caudal mesenteric vein to the liver. When the valve is closed. visceral and articular gout can be present in the same bird. 394 . including gout. gizzard erosions have been associated with naturally-occurring urolithiasis. and inflammatory products to the avian kidney. The avian caudal mesenteric vein drains the mesentery of the hindgut into the hepatic portal and/or the renal portal vein. However. and /or into the internal vertebral venous plexus within the vertebral canal. Gout Renal disease may lead to numerous other conditions. Gout may be only a nutritional disease in birds with hypovitaminosis A.the valves are open. obstructed ureters. Blood from the renal portal system can flow through the portal valve into the vena cava. Diffuse urate deposits on visceral surfaces do not occur in articular gout. blood flows into the parenchyma of the kidney. Clinically. persistent and progressive kidney damage is likely to occur. A potential consequence of the venous supply: Gastrointestinal ulcerations are reported in some animals with uraemia and advanced renal disease but are rarely mentioned concurrently in clinical reports of birds with kidney disorders. Gout reportedly may be caused by reduced excretion of urates or by increased dietary protein Dehydration. toxins. urate crystals precipitate. these connections with the renal portal system help explain the spread of neoplasia and infectious agents to other parts of the body. and following ureteral ligation and urolithiasis. Continuing damage Once renal damage occurs. As blood uric acid rises and exceeds its solubility (hyperuricemia). Articular gout results from accumulation of urates in the synovial capsules and tendon sheaths of the joints. Experimentally. Valve closure is inhibited by norepinephrine and epinephrine so that with “flight” there is available venous return to the heart. Although no refereed literature describes the post-treatment progression of renal lesions in living avian patients. Acetylcholine stimulates its closure. visceral gout has been induced in chickens fed excessive dietary calcium and a diet deficient in vitamin A. the author has observed from repeated biopsies that some mild renal lesions persist even if the patient is clinically normal or improved. which can further damage the kidneys or other organ systems. at least in chickens. In chickens. Visceral gout results secondarily from elevated plasma uric acid levels and its resultant deposition on visceral organs. enlargement. oocysts.and macrogamonts. Grossly. Chickens experimentally infected with E. distortion. developed a fairly consistent pattern and progression of renal disease. The typical lesions suggestive of bacterial nephritis include tubular dilatation and impaction with inflammatory cells. The tubules are often distended with endogenous developmental stages (micro. Histologic patterns are highly variable as some viruses directly affect the kidneys.Infectious diseases Bacterial: Certain patterns may be expected with bacterial nephritis. macrogametes) and maturing Emeria sp. Crowding likely favours transmission. Light microscopic identification of bacteria within renal tissue may be difficult as has been noted in dogs and swine with renal disease. such as pheasant coronavirus-associated nephritis. The prepatent period appears to range between species and has included 5 to 21 days. Staphylococcus aureus and Actinomyces pyogenes. is the most frequently reported avian renal parasite in those species and has been clearly associated with disease in some cases. Renal coccidiosis. As nephritis becomes chronic. In summary. Numerous viruses may infect and affect avian kidneys. oocysts are passed in faeces via the ureter and sporulate rapidly in the environment. bacterial nephritis is often a component of systemic infection and multiple organs may be involved. found predominately in some waterfowl and marine species. Most clinically affected species are young birds. tubular necrosis. and hemorrhagic foci. Lesions progressed to nephrotoxic nephritis and included tubular epithelial cell degeneration and necrosis with the formation of hyaline casts and eosinophilic material. Birds inoculated subcutaneously developed more severe renal lesions and these lesions were noted earlier than those exposed to bacteria Per Os. Using sterile collection and culture methods. 395 . Renal Eimeria sp. Viral: Viruses perhaps have the most varied effect on avian kidneys. Affected birds are typically emaciated and may have diarrhoea with or without blood. and interstitial fibrosis with mononuclear cell infiltration become evident. coli 01K67(B12)]. As is likely true of most viral and fungal renal diseases. Although transmission between different avian species is not clear. one study suggested that renal coccidia of geese do not infect ducks. are best identified in the kidney and provide an additional reason to save extra renal tissue (frozen and/or formalinised) for later testing. bacterial nephritis is definitively diagnosed by recovering bacterial organisms from affected kidneys. the kidneys are often enlarged with white to yellowish nodules containing urates and/or oocysts. Gross renal changes included congestion. Some viral infections. many of which are wild with concomitant intestinal parasites. coli [E. Parasitic: Renal coccidian Primary and secondary renal parasites have been noted throughout the avian literature and some contribute to significant morbidity and mortality. any septicaemia can potentially result in kidney infection and inflammation. while others like psittacine herpesvirus and polyomavirus damage renal tissue as a part of a more systemic process. such as the West Nile virus. cyst formation. especially those with concurrent immunosuppressive illness. Microsporidia Microsporidia (Encephalitozoon sp. Pulmonary cryptosporidiosis was also a common feature of the pullets. Several authors have hypothesized that urinary tract Cryptosporidium infection originates in the cloaca and retrogrades into the kidneys via the ureters. Sarcocystis organisms have also been noted histologically in the renal parenchyma of cockatiels. it has been directly associated with kidney lesions in a 4-month-old black-throated finch (Peophila cincta). Although relatively uncommon. Nephroblastomas are the most commonly reported avian renal tumours. depression. direct extension from the respiratory system (rather than primary renal invasion) is likely the cause of the necrotic fungal lesions in the kidneys. Cryptosporidium sp. Given the close association between the air sacs and kidneys. and adult laying hens. but again the significance is unclear.) have been reported in numerous avian species with variable effects on the kidney including a PBFD positive Eclectus parrot. Avian leucosis virus (ALV) can induce renal tumours in chickens. Fungal: Fungal nephritis is uncommonly reported in birds. and a double-yellow headed Amazon parrot. is associated with visceral gout most likely from partial ureteral obstruction. an 8-week-old Sonnarat’s jungle fowl (Gallus sonneratii). 3 species of lovebirds. Cryptosporidia Although renal cryptosporidiosis is infrequently reported.Sarcocystis Canaries (Serinus canaria) experimentally infected with Sarcocystis falcatula developed mild multifocal interstitial renal infiltrates and glomerular hypertrophy with mesangial hyperplasia that modestly progressed with duration of infection. There are likely many causes of renal tumours in birds but there is little information regarding definitive aetiologies. urinary tract cryptosporidiosis and associated disease seem to primarily be a concern in chickens. While ALV has been found in budgerigars with renal tumours. budgerigars. is susceptible to neoplastic conditions. a definitive 396 . Renal carcinomas are the most frequently reported tumours of the urinary system in non-domestic free-ranging and captive birds. Clinical signs ranged from acute death (finch and jungle fowl) to thinning. leg weakness and respiratory distress (4-month-old pullets and 4-day-old chicks) to slightly increased morbidity and mortality (adult chickens). just as with other animal tissue. Neoplasia The avian kidney. Four-day-old chickens co-infected with Marek’s disease virus have also been studied. Nephroblastomas and renal adenocarcinomas comprise the majority of kidney tumours in budgerigars. 4-month old pullets. Most (approximately 95%) of this volume is reabsorbed by tubular reabsorption. 397 . In addition. abdominal distension. enhanced tubular reabsorption. Other clinical signs may vary but often include diarrhoea. This likely result from a change in the rate of filtration and/or the rate of reabsorption. In addition to lameness and muscle atrophy. a decrease in the extracellular fluid volume using a haemorrhage model results in the release of AVT. The other effect of water overload and/or an increased extracellular fluid expansion results in an increased GFR to compensate. to hyperosmotic at 2–3 times the osmolality of plasma. ipsilateral osteopenia was noted in a cockatiel (Nymphicus hollandicus) with a renal adenocarcinoma. 5 REGULATION OF WATER EXCRETION The kidney filters a large volume of fluid daily with up to 11 times the total body water for a 100-g bird. with a range of less than 70% to more than 99% of the filtered volume. A common presentation with renal cancer is unilateral to bilateral leg weakness or paralysis and slight ataxia. The most common reason for AVT release is a rise in extracellular fluid osmolality. Dehydration also results in an increase in circulating levels of AVT. dyspnoea. The lumbar plexus lies dorsal to the cranial renal division while the sacral plexus runs through the middle division parenchyma. Additional neoplastic extension to the overlying spinal column may also result in nerve dysfunction. AVT is an 8-amino-acid peptide hormone that is released by the neurohypophysis. at approximately 40mmol/kg. As in mammals. For this reason. Studies using isolated nephrons or those using micropuncture techniques suggest that the proximal tubules are capable of reabsorbing 70% of the fluid volume. not seen with most spinal cord lesions. This reabsorption is dependant on active sodium reabsorption. the mechanism is not known. arginine vasotocin (AVT). This is important as these nephrons lack a loop of Henle and therefore the ability to concentrate urine. indirectly. any parenchymal inflammation or pressure on. reabsorption depends on a counter current multiplier system of the loop of Henle that is produced by an osmolality gradient in the medullary region. and weight loss. These changes can result from the increased levels of the circulating antidiuretic hormone. Because of this close association. but not bicarbonate.association has not been made. The reduction of GFR may result from constriction of afferent arterioles that causes a reduction or even a cessation of filtration in some or all nephrons. Peripheral neural compression should result in peripheral neuropathy with eventual loss of the withdrawal reflex. Reptilian or cortical nephrons are the most sensitive to AVT. which leads to reduced urine flow because of reduced filtration and also. or from within. as occurs in mammals. the kidney can potentially result in nerve dysfunction and resultant lameness. the urine can range from dilute. However. Birds also have the ability to concentrate their urine by tubular reabsorption. Associated with the rise in AVT is a reduction of the glomerular filtration rate (GFR). ‘albumenous’ casts in renal tubules. but it may involve tubular reabsorption and/or reduction of filtration by cortical nephrons. as they run through this osmolality gradient.. the ‘urine’ present in a dropping is not the same urine produced from the kidneys. as noted in Amazona spp. inflammation) may alter urine production and composition.against a concentration gradient while having a low permeability to water and net water transport. In birds. This results in reabsorption of ions in the filtrate as it moves through this TAL portion of the nephron. This produces the highest osmolality at the tip of the loop and the development of a gradient outward from there. This latter action would result in an increased osmolality gradient in the medullary region. hematuria may be noted with renal disease but should be carefully differentiated from bleeding originating from the gastrointestinal and reproductive tracts. the thick ascending limb (TAL) allows for the transport of Na+ and Cl. gastrointestinal bleeding. diseases of the lower intestine (eg. In short. Protein and cellular casts. parrots with lead intoxication and in other species with differing disorders. hyaline casts in kidney sections or renal tubules. Further concentration of the urine may occur by retroperistalsis up into the coprodeum and the large intestine.but has low water permeability. many of which had interstial nephritis. 6 DIAGNOSTIC TESTS Urinalysis Biochemical and cytological sediment analysis of avian urine has been advocated as potentially useful in diagnosing avian renal disease. eosinophilic granular 398 . Exactly how AVT alters this system remains unknown. However. specially designed cannulae can be inserted into the cloaca for collection of ureteral urine. while birds greater than 500 g concentrate to 600–700 mmol/kg. Ureteral urine may be refluxed orad into the colon up to the ceca where water. Additionally. The net effect is that birds are able to concentrate their urine often at 2–3 times that of plasma. Once emptied of faeces. and viral nephropathies may be more frequently seen associated with hematuria in birds. White blood cells were seen in 45% of urine sediment from pigeons with paratyphus. Sediment analysis may be complicated by anatomical factors. Collection: True urine can be collected in birds with some difficulty.The thin descending limb (TDL) is permeable to Na+ and Cl. Casts: Urinary casts represent cellular and/or acellular material sloughed from the inner lining of various renal tubules. bacterial. Urine may be mixed with faeces when voiding. Water permeability of the collecting ducts. may or may not be related to renal disease. and sometimes electrolyte. neoplastic. Toxic. reabsorption takes place. A foley catheter was used in chickens. produces the concentration of urine over plasma. which are able to reabsorb water. Hemoglobinuria. The urinary concentrating ability generally varies inversely with body mass so that small birds (10–25 g) typically concentrate to 1000 mmol/kg. as noted in diving ducks. Because ostriches can eliminate urinary waste separate from faeces. and ketones in all ostriches. Osmolality and specific gravity: Avian urine is typically isosmotic with its osmolality maximally increased to 2. phosphate.1 to 9. Once the egg is laid or calcium is no longer being deposited. Because of inconsistent results for other analytes in other species. difficulty in obtaining ureteral urine.0-2. E. the author (M. Urinary casts may be a sign of renal disease but there is little information correlating them with any type of renal disease in birds. urine osmolality significantly increased up to 3 times control levels in post-flight and dehydrated pigeons.casts in renal tubules.007) in Salmonella typhimurium-infected pigeons. and total osmolality than found in ureteral samples. these values may not apply to most other birds. GAVAERT et al.7) in egg laying female birds during calcium deposition. Few studies mention test strips for use in avian urinalyses. noted consistent polyuria and hyposthenuria (60% had specific gravity below 1. 31/35 (89%) and 35/35 (100%) of the urine samples were positive for nitrite and protein. in house sparrows undergoing trials with the antidiuretic arginine vasotocin. Urine electrolytes and chemistries can be collected. may drop urine pH to 4. S. and insignificantly changed. 399 .6.1.4. many of which had interstitial nephritis. but there is limited information on their interpretation. potassium. One study noted that in normal and dehydrated starlings (Sturnus vulgaris). The urine pH may be acid (down to 4. bilirubin. eosinophilic casts in renal collecting tubules. urinary chemistries might be useful in detecting kidney damage. One study evaluated commercial urine dipsticks (Combur-9 stix [Boehringer Mannheim]) on normal urine of 35 ostriches (Struthio camelus). respectively. No association with renal disease was made. Urine pH: Urine pH is highly variable in birds. This is minimal compared to mammals that can concentrate urine osmolaltiy 25 to 30 times that of plasma. cloacal urine contained significantly higher concentrations of magnesium. In the study. limited critical studies.5 times that of plasma. There is limited information on urine specific gravity or osmolality in avian health or disease. eosinophilic tubular casts. Chicken urine reportedly contains non-uric acid chromagen. The urine chemistry strips were negative for glucose. and clinical experience. urinary pH may climb to 8. Normal ostriches have a urine pH range of 6. urobilinogen. Urine chemistries and electrolytes: Standard mammalian dipsticks may be used but not all components are applicable to avian urine. Non-protein chromogens are known to interfere with refractometric and chemical measurement of plasma proteins and may also apply to avian urine sampling. In a separate evaluation.) believes that chemistry strips currently available have limited value for avian urinalyses. Male birds have an approximate urine pH of 6.0. and epithelial casts in urine samples have been identified in various species. LUMEIJ suggested that because renal intracellular enzymes are likely voided in the urine.7. with a mean of 7. Urine sodium and potassium were measured. Hypoxia. As viewed with a lateral radiograph.09 mg/ml in dogs and humans). However. The reflux of urine into the cloaca may be a mechanism to recover some of the urinary protein as cloacally voided fluid contains very little protein compared to ureteral samples. is found in both the liquid urine and uric acid spheres in chickens. Indirect methods such as positive contrast radiography of the alimentary tract may be helpful in outlining renal masses. Ultrasound: Ultrasonographic imaging of normal avian kidneys is difficult because of the surrounding air sacs (ventrally) and bone (dorsally and laterally). A lateral view is the best method to view the kidneys radiographically. Nephrocalcinosis was detected radiographically in ostriches and appeared as multiple radio-opacities throughout the renal parenchyma.Electrophoresis Plasma protein electrophoresis: Properly determined hypoalbuminemia (via plasma electrophoresis) is not reported in confirmed active cases of avian renal disease. abnormalities. imaging of the avian renal system may be difficult. and inflammatory nephromegaly (11). In one study of 386 mixed bird species that underwent ultrasonographic evaluation of the urogenital tract. In these abnormal situations. ultrasonography can serve as a non-invasive and safe means to evaluate coelomic structures such as the kidneys. cancer (12). Because the renal silhouettes are superimposed on a lateral view of the abdomen. magnetic resonance imaging. Renal density and gross size changes may indicate renal disease. Because the avian kidney is surrounded by bone dorsally and air sacs ventrally. nuclear scintigraphy. Sonographic imaging of the normal kidney was deemed not possible. Urinary protein electrophresis: Avian urine normally contains a large amount of protein (average of 5 mg/ml up to 15 mg/ml). especially when compared to that of mammals (< 0. the absence of the normal dorsal diverticulum of the abdominal air sac (dorsal to the kidney and ventral to the synsacrum) may indicate renal enlargement. Some disease conditions that either obliterate the air sacs or result in fluid accumulation in the coelomic cavity may actually improve renal ultrasonographic imaging. such as renal cysts (6). and computed tomography can be used to ‘image’ the avian kidneys. preventing aggregation and renal tubular blockage. Radiographically visible renomegally was noted in a salmon-crested cockatoo (Cacatua moluccensis) with chronic interstitial nephritis and calcification as the result of hypervitaminosis D3. among other proteins. it is possible that birds may develop low albumin/protein with some kidney disorders. 400 . ultrasound. an oblique view may also be used to distinguish each kidney. Improper positioning can artifactually change the appearance of this air-filled diverticulum. Biochemically determined hypoalbuminemia has been noted in some active avian renal disease cases. Proteinuria is likely necessary to maintain the excreted uric acid-containing spheres in a colloidal suspension. Serum albumin. Imaging Radiography: Plain and contrast radiography. were identified in only 29 patients. Uric acid crystals are easily confirmed using microscopy or the murexide test. shape. 7. depending on the species and physical condition of the bird. 7 WATER DEPRIVATION TESTING Water deprivation testing is considered when attempting to rule out unknown causes of polyuria/polydipsia (PU/PD) including central and nephrogenic diabetes insipidus. After 2 to 5 minutes. A presumptive diagnosis is based on whether birds can concentrate their urine. LUMEIJ reports using organic iodine compounds (Urographine 76) given IV at 2 mg/kg in the basilic vein. endocrine (diabetes mellitus). and metabolic (hypercalcemia) diseases. 2. Some of the many causes of PU/PD in birds to consider include organic (liver. cardiac. Intravenous excretory urography may have some limited uses in a clinical setting. This technique should not be used in birds with severe renal compromise. etc). chalky substance in joints (articular) or on tissue surfaces (visceral). The needle-shaped crystals are easy to identify on direct and stained smears. Squib Diagnostics. the cloaca will be outlined. As a normal response. Articular gout material may be recovered using fine needle aspiration. intestine. The organic iodine can be visualized radiographically in the heart and pulmonary artery within 10 seconds and outlining the kidneys and ureters 20 to 50 seconds later. 401 .Intravenous excretory urography: Intravenous excretory urography has been described in birds as a method to gain information on kidney size. and function. Cytologically. Birds with diabetes insipidus become dehydrated (as supported by plasma variables) but maintain dilute urine (low specific gravity and osmolality). The avian patient is caged with no food or water for the duration of the test. USA) to evaluate the ureters post-ureterotomy in a double-yellowheaded Amazon parrot (Amazona ochrocephala). kidney. Smaller birds should be evaluated at more frequent intervals. Renografin-76. NJ. Princeton. ‘gouty’ material typically presents as uric acid crystals surrounded by a pyogranulomatous infiltrate. urates can be seen in alcohol fixed tissue using Gomori’s methenamine silver impregnation technique. and psychogenic polydipsia. urates will dissolve in formalin and therefore the crystalline form will not be seen on conventionally fixed tissue. usually without organisms. However. Radiographic images taken at 1. some birds such as European starlings may become distressed within 24 hours. Hereditary diabetes insipidus has been described in a strain of chickens that produce low osmolaltiy urine and maintain high circulating level of AVT. and 10 minutes post-injection were used to evaluate ureter peristaltic movement and size. Due to their water-soluble nature. Evaluate both blood and urine parameters every 3 to 24 hours for 12 to 48 hours. Identifying uric acid crystals Gout results when uric acid precipitates out as a solid. DENNIS and BENNETT successfully used a water-soluble iodinated contrast agent (400 mg/kg IV in right medial metatarsal vein. 8 AVIAN RENAL DISEASE TREATMENT Therapeutic considerations Treatment options for renal disorders in birds depend upon the cause and type of kidney disease and secondary complications present. relieving a potential ureteral obstruction. culture media (organism recovery).) has also used minor surgery in articular gout cases. consider biopsy. kidneys. renal tissues can be stored in 10% formalin for light microscopy. 402 . If available. Most renal disease patients are medically managed. additional tissue may be stored in glutaraldehyde (electron microscopy). It is often useful to combine the pathologist’s interpretation with the veterinarian’s first-hand knowledge. Renal histologic lesions are rarely pathognomonic for a specific disease process. S. avian kidneys are difficult to remove surgically. For the most part. However. and alcohol (visualizing uric acid crystals) or frozen (PCR studies). The close associations with the lumbar and sacral plexuses and extensive vascular network surrounding the kidneys lead to the high probability of significant haemorrhage expected during surgery and possible neurologic damage. Renal stones were successfully removed via extracorporeal shock wave lithotripsy in a Magellanic penguin (Spheniscus megallanicus) and ureteral stones successfully removed from a 21-year-old male double-yellow-headed Amazon parrot. as kidney surgery is difficult and often not needed. Before a renal biopsy is performed. A renal biopsy is most frequently performed during endoscopic examination of the coelomic cavity and specifically. therapeutic surgery (including endoscopic biopsy) for superficial renal lesions and the ureters may sometimes be useful. Because of the location within the renal fossae. most ‘surgical’ renal disease cases are managed medically. the only way to definitively diagnose avian renal disease and specific pathologic patterns is with a kidney biopsy and histopathologic evaluation. Small incisions are made over the gouty lesions and the thick material expressed. primarily via endoscopy. The author (M. the cost: benefit of the surgical procedure versus conservative therapy must be considered as many birds have compromised health. Several methods of renal biopsy.Biopsy When history. Bandaging is often required to stem bleeding and prevent secondary infection. and detailed accounts of avian kidney anatomy and physiology have been previously discussed. and/or laboratory abnormalities support the presence of renal disease. Cloacaliths and other masses within the cloaca may be easily removed. Currently. physical examination. especially if they have kidney disease. Many different diseases cause similar renal lesions and different pathologists may make differing morphologic diagnoses on the same renal tissue. Given the concern of serious haemorrhage. Anaesthesia is recommended as this can be quite painful. E. A few accounts of therapeutic renal surgery exist. ) typically diureses ill and severely hyperuricemic renal disease patients. Clinical consideration regarding potential antimicrobialinduced toxicities is important. Until acid/base and electrolyte disorders are better evaluated in birds with renal disease. The IV and IO routes are most appropriate for critically ill patients. such as Lactated Ringer’s Solution (LRS). and/or supportive histopathology). replace fluid losses. Because of the renal portal system and possible shunting of blood from the intestines directly to the kidneys. and/or induce diuresis as needed. The estimated daily fluid requirement for most birds is 40-60 ml/kg/day. Antibiotics Antibiotics are indicated in patients with known or suspected bacterial nephritis. more active. ovarian. salpingitis. maintaining hydration is important in birds with most kidney disorders. Oral fluids are reserved for stable patients with mild dehydration that have normal gastrointestinal function and are often contraindicated in critically ill birds. Subcutaneous fluids are not adequate to rehydrate patients with severe dehydration. Anuric and oliguric patients should be diuresed. balanced electrolyte solutions. E. or intraosseously (IO). even with most forms of renal disease. etc). extension from local tissues (peritonitis. oophoritis. Acid/base and electrolyte disorders may likely be present in birds with renal disease. LUMEIJ recommends that 10% of the bird’s body weight should be given in fluids when the patient is in renal failure.Diuresis and fluid therapy As in other animals with renal disease. Fluid therapy (combined with other medications if needed) is generally continued until the blood uric acid level drops to either normal or mildly elevated levels (10-20 mg/dl) and the bird shows signs of improvement (eating. shock. salpinx. IO. Clinically. providing parenteral fluids often induces diuresis in birds. alimentary tract organisms may contribute to kidney disease and should be considered when using antimicrobial therapy. should be used to maintain hydration. S. etc). identified during kidney biopsy sampling) or a suspected infectious agent (blood. Although manitol and lasix have been recommended to induce diuresis in birds. and haematogenously. Drug choices are based on an isolated renal organism (ie. The author (M. the author will typically give 50-100 ml/kg of fluid q12h via the SC. or hypothermia. 403 . Warmed fluids are given either with food (tube/ syringe fed). or combination routes. IV. ’Severe hyperuricemia’ is identified when one or more of the following conditions are met in clinically ill non-carnivorous and appropriately fasted carnivorous birds: • uric acid levels exceed 30 mg/dl • uric acid levels are elevated (> 10 mg/dl for most species) and rising over a period of several days (even if below 30 mg/dl) • there is evidence of rapidly progressive articular or visceral gout Depending on the patient’s condition. or cloacal/faecal cultures. these drugs are poorly studied in avian species. SC. IV. Bacterial renal infections in birds may result from an ascending ureteritis. In humans. cephalosporin. S. For these reasons. respectively. but had no significant effect on plasma uric acid concentrations. colchicine reversibly inhibits xanthine dehydrogenase (compared to a ‘pseudo-reversal’ with allopurinol). renal fibrosis and amyloidosis Allopurinol: Allopurinol’s main action is to decrease uric acid production. Although the long-term effects are not clear. which is required to convert hypoxanthine to xanthine and subsequently to uric acid. colchicine has been used to treat amyloidosis and hepatic fibrosis. In small animals.) uses allopurinol as a first-line drug to lower uric acid when fluid therapy and diet modification alone are not sufficient or when hyperuricemia is severe. Colchicine: Theoretically. colchicine blocks the synthesis and secretion of serum amyloid A and decreases the formation and increases the breakdown of collagen. colchicine can reduce serum uric acid levels in birds and be used to control hyperuricemia. With the exception of red-tailed hawks. and toxicities of many antimicrobials are poorly defined in most bird species. elimination. flouroquinolone. Allopurinol has been specifically shown to prevent renal synthesis of urates and allow the excretion of unchanged xanthine. allopurinol given to chickens increases oxidative activity by lowering plasma uric acid.) has also used colchicine successfully to reduce renal (and hepatic) fibrosis. an important avian antioxidant. trimethoprim/sulfonamide. Those drugs with known potential nephrotoxicity should be used cautiously in birds with renal impairment. E. Allopurinol has been shown to be toxic to red-tailed hawks (Buteo jamaicensis) at 50 mg/kg PO q24h or at higher doses. colchicine is best known for its anti-gout activity. only aminoglycosides have been consistently and definitively associated with renal disease in birds. allopurinol inhibits xanthine oxidase. allopurinol use is reported to be non-toxic in birds (in studied chickens).The distribution. The author (M. S. Clinical use of colchicine suggests possible benefit in reducing hyperuricemia in birds with renal disease. Allopurinol administered at 25 mg/kg PO q24h was shown to be safe. Antimicrobials that reach high concentrations in the renal tissue and urine without inducing toxicity should be chosen preferentially and used cautiously in kidney disease patients. E. colchicine may be used in birds with amyloidosis. Colchicine prevents the progression of renal disease in humans with familial Mediterranean fever. Allopurinol and colchicine are generally well tolerated when given together. Toxicity was attributed to oxypurinol. a disease of recurring fever often complicated by amyloidosis. Clinical and experimental data show decreased plasma/serum and/or urinary uric acid levels in birds treated with allopurinol. Although mammalian literature warns of potential nephrotoxicity with amphotericin B. and tetracycline use. The author uses colchicine as a second-line drug to reduce hyperuricemia and a primary medication for histologically confirmed tissue fibrosis. the active metabolite of allopurinol. Specifically. The author (M. If diagnosed pre-mortem. 404 . Managing hyperuricemia. In chicken livers. lipid-stabilizing and anti-neoplastic effects. EPA also serves as a substrate for the formation of vasodilatory prostaglandin/cyclins (PGI/ PGE) and their respective products (PGI2 /PGE2 and PGI3 /PGE3). respectively and therefore have different N-3 FA compositions. As with arachidonic acid. most animals readily convert omega-6 fatty acids (N-6 FA) to arachidonic acid and subsequently eicosanoids (prostaglandins. These vasodilatory prostaglandin/cyclins increase renal blood flow and single nephron GFR. Nutritional supplementation Treatment: omega-3 fatty acids: Omega-3 fatty acids (N-3 FA) have gained popularity for their anti-inflammatory. birds should be fed diets appropriate for their species. renal protective properties. In mammals. all of which have similar biologic potency. but the identification of hypoproteinemia and its association with renal disease is unclear. This pathway decreases the release of arachidonic acid and. A study with pigeons and red-tailed hawks concluded that ‘urate oxidase is much more effective compared with allopurinol’ but further evaluation is needed. Protein: The question of dietary protein restriction in the face of renal disease remains controversial. Careful supervision and adequate calories are urged in the human literature. docosohexaenoic (DHA). Current human and veterinary literature both supports and refutes protein restriction in patients with renal disease. subsequently. Studies evaluating N-3 FA in mammals serve as the basis for potential treatment value in birds with selected renal disease. Management of hypoproteinemia may also be important in birds with renal disease. At least in humans. In contrast. N-3 FA are those rich in eicosapentaenoic (EPA). Avian kidney disease patients should be fed a well-balanced diet appropriate to the species. Dietary modification As a general note. N-3 FA can significantly reduce thromboxane A2 (TXA) synthesis in platelets and glomerular cells and increase production of vasodilatory prostaglandins. and/or linolenic acid. Birds fed protein-restricted diets should be carefully monitored. the cyclooxygenase-mediated synthesis of TXA. At this time.Urate oxidase: Urate oxidase has also recently been discussed as an alternative method to manage hyperuricemia in birds. only anecdotal information exists regarding use of N-3 FA in birds with renal disease. N-3 FA partially substitutes EPA and DHA acid for arachidonic acid in membrane phospholipid. A safe recommendation is that birds with hyperuricemia and/or gout should not consume diets with protein levels greater than is normal for the species. 405 . and other potential qualities. Flax seed and menhaden (cold water plankton-feeding fish) oils contain predominately linolenic acid and EPA and DHA. which the kidneys can more easily clear than uric acid. urate oxidase is reported to degrade the excess of uric acid to allantoin. Renal disease patients should be monitored regularly for weight loss. TXA). In general.0 mg/kg PO q 12 h) together to manage histologically confirmed glomerulopathies in avian patients. Non-steroidal anti-inflammatories Non-steroidal anti-inflammatory drugs (NSAIDS) are frequently discussed for use in human and animal renal disease patients. NSAIDS such as aspirin and ibuprofen are non-specific cyclo-oxygenase inhibitors. Although there are limited avian studies. NSAID therapy have not been studied in birds with renal disease. Supplementation with 160 mg/kg of vitamin E (dl-�-tocopheryl acetate) was shown to prevent loss of �-tocopherol in tissues and normalize or increase resistance to lipid peroxidation in chickens fed a commercial diet supplemented with 3% tuna oil (N-3 FA). low dose aspirin therapy has been suggested to reduce platelet aggregation and subsequent thromboembolism and to minimize glomerular inflammation for mammalian patients with some glomerulopathies. A dietary N-6 FA: N-3 FA ranging from 5:1 to 15:1 has been proposed as desirable for dogs and cats with renal disease. Diclofenac-treated livestock caused several Gyps spp. In birds with suspected hypovitaminosis A and renal disease. Consequently. In such situations. 406 . Supplementing the diet with N-3 FA increases the requirements for dietary vitamin E. Vitamin A: Hypovitaminosis A is a reported cause of renal failure and results from metaplasia of the ureters leading to hyperkeratinization. while therapeutic administration of diclofenac has been implicated in the decline of the Oriental whitebacked vulture (Gyps bengalensis). the beneficial effects of low dose.Specific ‘toxicities’ associated with N-3 FA supplementation are poorly described but some potential adverse effects may occur. have been shown to attenuate renal dysfunction/damage.22 cc/kg body weight. decreased mucin production. and impaction. Flunixin meglumine (Banamine®) induced glomerular lesions in bobwhite quail (Colinus virginianus) in proportion with the dose. or specific. More specific NSAIDS. some believe that the appropriate N-6 to N-3 FA ratio is more important to inhibiting detrimental eicosanoid synthesis from arachidonic acid than is the absolute amount of N-3 FA. Aspirin has been associated with significant inhibition of prostaglandin synthesis in Japanese quail (Coturnix japonica) and with temporary diuresis in Pekin ducks (Aix galericulata) when administered IV.51. The author has successfully used supplements containing N-6 FA: N-3 FA of 4-5:1 to 1:3 (0. Unfortunately. appropriate diet modification and short-term parenteral vitamin A is a logical therapeutic component. vultures to die of renal failure and gout. most NSAIDS are eliminated by renal clearance and should be used with caution. Low doses of aspirin may actually inhibit platelet cyclo-oxygenase production but allow beneficial (vasodilatory) prostacyclin formation and may be safe. such as thromboxane synthetase inhibitors. PO. as they have been associated with a variety of renal lesions in birds and mammals. Although specific doses have not been established. the author gives a single IM vitamin A injection at the beginning of the therapy and recommends correcting the patient’s diet to improve long-term nutritional status. SID) combined with low-dose aspirin (0. N3-FA can be given chronically if needed. Give vitamin A if hypovitaminosis 407 . For renal fibrosis. and vitamin A supplementation may also be indicated. For colour variety psittacine birds. Otherwise use colchicine for 6-12 months. Parenteral vit A may also be provided. If after 3-6 months all signs of renal disease are gone.5-1. Articular gout can be treated with colchicine and allopurinol together until all signs of gout and hyperuricemia have resolved. E. remove/control any source of infection/inflammation. accurate renal histologic diagnosis (if available). or until laboratory abnormalities normalize. infectious diseases. S. Confirmed glomerular disorders in birds without an obvious underlying disease may be managed in some cases with low-dose aspirin and N-3 FA supplementation. hyperuricemia. Treatment summary Treatments of avian renal disease should be individualized according to the patient’s needs.Even with the noted toxicities and lack of therapeutic studies in birds. and possibly other NSAIDS.0 mg/ kg PO q12h) combined with N-3 FA supplementation is safe and may be effective at reducing the severity of some forms of avian renal disease. Otherwise. low dose aspirin. Consider N3-FA supplementation. especially glomerular disorders. Anti-neoplastic treatment of certain avian renal tumourss may be indicated and should be considered. Consider diagnosing the cause of probable underlying renal disease and manage appropriately. Nephrosis may best be managed by removing known nephrotoxins and addressing secondary complications. Antibacterials should also be considered when concurrent colitis is present. N3-FA may also be beneficial. in part. and anorexia. If identifiable. Aspirin (and N-3 FA) therapy can be used chronically. can be beneficial in treating avian kidney disease patients. low dose-aspirin (0. In the author’s experience (M. Provide antibiotics for a minimum of 6 weeks. concurrent disorders. providing a balanced diet. pellets (< 50% of total diet) can be cautiously added to diet. discontinue pellets and change diet over to whole grains.). Discontinue use when evidence of renal disease is gone or the disorder is satisfactorily managed. fibrosis. Specifically identifying and managing underlying diseases that may be concurrently present may best control glomerulopathies. bactericidal antibiotics that reach high kidney concentrations and are non-nephrotoxic. Such secondary complication of any renal disease may include dehydration. and client considerations. Nutritional management such as weight loss. on culture and sensitivity results when available. seeds. use colchicine until histologic fibrosis resolves. suspected bacterialinduced nephritis should be treated with broad-spectrum. and vegetables as is appropriate for the species. Dietary-induced renal diseases can be managed with diet change or supplementation depending on the aetiology. Treatment of bacterial nephritis with appropriate antibiotics should be based. fruits. Texas. AUTHORS ADDRESS MS Echols DVM. Dipl ABVP (Avian) Westgate Pet and Bird Hospital.com 408 . Based on the author’s experience. giving a good prognosis for long-term health to the individual patient. etc should be managed as needed. unacceptable weight loss. Austin. USA Email:
[email protected] is suspected. dehydration. N3-FA may be beneficial. Prognosis There are limited studies that estimate the outcome of selected avian renal disorders. several forms of renal disease can be successfully managed. Combination therapy should be considered when two or more histologic renal lesions are present. and some resolved. Again. Articular gout lesions may also be surgically opened and expressed to speed removal of uric acid crystal accumulation. Secondary infections. Use aggressive fluid therapy if articular or visceral gout is accumulating rapidly. In 2 diagnostic tables the most common problems in small passerines will be covered including suggestions for additional diagnostic tests. The Netherlands PASSERINE AND SOFTBILL MEDICINE AND SURGERY G. including diagnosis and treatment.8 to 1350 grams. and therapeutics will be summarized. feeding. Special attention is paid to the diagnostic possibilities and infectious and parasitic diseases.Diseases . Passerines (songbirds) are regularly presented for veterinary care as aviculturists recognize that a visit to an avian veterinarian can result in a successful medical treatment. Exotic Animal and Wildlife Section Utrecht University. For an optimal approach of the veterinary problems.Department of Pathobiology. Dorrestein. Pet Avian. Supportive care and measures to minimize stress are often needed to maintain the host’s defence mechanisms. diagnostic procedures and common diseases and their treatment will be discussed based on own experience and recent publications. Diseases in these birds are very much influenced by nutrition. DVM. PhD KEYWORDS Passerines . housing and husbandry of these species. housing and stress. even in these oftentiny patients. management and diseases of the small passerines (canaries and tropical finches). with body weights ranging from 4. The aviculture. Toucans and mynahs are often grouped together. 409 . 1 INTRODUCTION Many of the veterinarians will be relatively unfamiliar with the passerines.Treatment ABSTRACT In a short overview basic information is presented of housing. blood chemistry. but are from different families. M.Husbandry . The order Passeriformes contains over 5700 species. In table’s basic haematology. it is a must to become familiar with aviculture. In the breeding-season the birds are mostly kept in couples in small box type cages (50x40x40 cm). their colours.). the housing depends on the season. eg. The weanlings are 410 . The canary (Serinus canaria) are bred and kept for their song (e. largest: Java rice sparrow 20 g. the border fancy and the Norwich).1 Housing The small Passeriformes are kept in captivity as individual pet birds and as flocks in aviaries. In breeding aviaries located in a garden house or in a loft.g. The birds are in the aviary all year with a shed for shelter and a fly pen outside. i. In the latter. is less dense and species specific diseases are restricted to only a few of the occupants of the aviary. for breeding and selecting.g. In mixed aviaries. presence of nest and nesting material.e. Nowadays canaries are mostly bred indoors. Disease control in planted aviaries can give difficulties related to controlling microorganisms and in medicating individual birds.. These fanciers often go to shows and competitions and there is exchange of birds (and possibly pathogens). large numbers of the same species are kept. the population. enough water and food. 2. There are several hundred species of finches and they have a worldwide distribution. an accepted partner. There is a fair amount of size disparity between the common finch pets (smallest: Gold-breasted waxbill 7 g. Cochlosomosis and Campylobacter infections. This gives special problems because they can be carriers of diseases that can kill the foster fledglings. zebra finch (Poephila guttata). consisting of individuals or couples of different species. The former are usually located outside and different species are kept in it together. Lady Gouldian (Chloebia gouldiae) and parrot finch (Erythrura psittacea) are kept for breeding but also as ornamental birds. The more domesticated species are bred in captivity since many decades.2 BIOLOGY AND HUSBANDRY 2.25 g. or their build and shape (e.1 Species The most common representatives of the passerines in captivity are canaries and finches. Normally canaries will start breeding when the following conditions are met: maturity and good health. the Harzer and the Spanish timbrado). mostly for ornamental purposes. Their weight is 15 . mostly indoors. The sexes are alike and their lifespan is 6-16 years. however. Planted aviaries are popular for these passerines because the vegetation provides observers a more natural view of a bird’s behaviour. These plants. Two types of aviaries can be distinguished: mixed ornamental aviaries and breeding aviaries. a minimum temperature and photoperiodic stimulation. Bengalese (or society finch (Lonchura striata domestica) and zebra finches are used as fosters for breeding Australian finches. Most common finches belonging to the family Fringillidae and Estrildidae. Normally the fancier breeds two to three rounds. are often necessary to get breeding results. a minimum length of the day. 2 Diet and Husbandry Dietary and husbandry requirements are diverse. and strong lighting in combination with a magnification device will greatly facilitate any examination of the tiny birds.housed in communal flights with or without outside quarters. Attention should be paid to the droppings. When handling the birds. Examination of the cage or aviary can provide a great deal of useful information. especially in flocks. The majority of commercially available passerine diets are seed mixes and therefore multi-deficient. available phosphorus. vitamin B12 and choline. The birds can acclimate then to their new surroundings and fresh stool for examination is likely to be produced. age. Singing canaries are housed individually in small sing-cages (21x20x15 cm) for more then 5 months to be trained and to enter the singing competitions. The basic approach like in parrot-medicine will lead to proper diagnosis and treatment. these procedures should be followed by a diagnostic necropsy. vitamins A.1 Clinical diagnostics The history should include information on the species. sodium. Some species adapt readily to commercially available diets. the feed dishes. selenium. The main clinical diagnostic procedures for these small birds are history. symptoms. insectivorous. even before the history is taken. Most breeders of passerines bring in their birds in transport boxes or cages. riboflavin. 3. available niacin. E and K. 2. iron. carnivorous. examination of the cage. Birds should be put in a birdcage as soon after arrival as possible. calcium. diet and housing. The breeding aviaries are relatively easy to clean. In many cases. Larger passerines like mynahs are more expensive. and the floor. omnivorous or. A careful history will provide much information needed to arrive at a diagnosis. keep the windows closed! 411 . zinc. Common nutrient deficiencies of seeds include lysine. Veterinary care in these species is frequently directed toward appropriate preventive husbandry measures and approaching medical problems from a flock perspective. manganese. while others may require live food. Transport in their own cage is recommended whenever possible. “Light out/perches out” catching techniques are almost mandatory. 3 DIAGNOSTIC PROCEDURES Diagnostic and treatment options in passerines may be limited by owners’ financial constraints and difficulties in collecting samples from small birds. The inadequacies of these seeds for growth apply generally for reproduction and. physical exam and limited clinical procedures. iodine. to a lesser extent. while others species are nectivorous. pantothenic acid. In the winter season (resting season) the males and females are housed as separate groups in pens. for maintenance of adults. The composition of the basic diet will be affected by the species of bird in question. frugivorous. but are beyond this presentation. Most common passerine species are primarily seed eating or granivorous and have been domesticated for centuries. D3. Do not interfere with the movements of the sternum. blood can be collected from the right jugular vein.g. but also with some practice from small finches. serologic. The usual physical examination is performed as for any other bird. but the container must be weighed or tarred. One drop is used for a blood smear. and histopathologic examinations and immunodiagnostic techniques are special techniques to come to a diagnosis. Routine diagnostic procedures also include the following: Faecal examination. and contact or impression smears from a freshly cut surface of liver.g. Crop swabs. Special attention should be paid to the state of moult. the abdomen (blow the feathers apart and look for an enlarged liver and dilatation of the gastrointestinal tract). The necropsy is also the ultimate method to confirm a diagnosis. Coccidia. Normal haematological and serum biochemical references are presented in table 1.g. For additional information in small individual passerine birds. Crop swabs are essential for the diagnosis of trichomoniasis and crop candidiasis. Giardia spp) are found with direct wet preparations or after flotation techniques. Campylobacter jejuni) can be found in stained faecal smears. but nevertheless very important. Yeasts and protozoal cysts (e. and the skin (search for pox lesions and parasites). - 3. scrapings from the mucosa of the crop. proventriculus. present. and any altered tissues. Bacteriologic. but are more often seen in wild-caught mynahs and insectivorous passerines. for the type of anaemia. Listen for respiratory sounds. Routine microbiologic aerobic cultures should be negative and microaerophylic strains (e. The smears are stained routinely with Romanowsky stains (e. The following procedures can provide much additional information during the necropsy: direct wet preparations of the gut contents and of the coating of the serosae. Because passerines are not considered to have a normal gut-flora. are excreted mainly between 2 pm and darkness.g. spleen. Blood samples. which are common in small passerines. Total protein (TP) determinations provide a very significant diagnostic measure. mycologic. duodenum. no bacteria or other microorganisms should be found in stained faecal smears.2 The diagnostic necropsy A necropsy should always be performed on birds that die from unknown causes so that flaws in management can be rectified and a possible epidemic can be forestalled. Giemsa) or “Quick” stains (e. which can be examined for blood parasites and if. The diagnosis of cochlosomosis in Society finches or Australian finches can only be made in direct wet mounts of fresh and warm stool without dilution. and rectum. which could kill the patient. any reading less than 35% indicates anaemia. virologic. From lager birds. the pectoral muscle mass (chronic or acute problem). DiffQuick) and searched microscopically (cytology) with oil immersion. Most gram scales can provide an accurate weight if the finch is contained in a paper box or bag. 412 .The physical examination and clinical procedures are limited in the smaller passerines. blood can be collected in heparinised capillary tubes after puncturing the medial metatarsal vein. Helminthic infections are very rare indeed in small passerines. lungs. The packed cell volume (PCV) normally ranges from 40% to 55%. Because the elimination is so rapid. or by placement of a capsule or granules in the centre of a grape or raisin. the injection site should be located in the caudal third of the breast muscle. Application via the drinking water is a common route of drug administration in birds. which will result in a more even distributed blood levels over a period of 24 hours. In many disease cases and after adding drugs to the water the water intake will be reduced substantially. Oral administration in larger passerines can be given directly. This change of day-night rhythm. For the different dosage rates see the different formularies. Medications can be easily administered by several methods.4 TREATMENTS Compared to the other orders of the class Aves passerines. by metal or rubber gavage tube. For parenteral administrations (intramuscular or subcutaneous) a 27-gauge needle is required. including mynahs. Administration of medication in the drinking water is especially not recommended in soft bills and fruit-eating finches because they generally do not consume large quantities of water. Treatment of skin or leg lesions with ophthalmic ointments often results in intoxication and the death of the patient. to achieve better therapeutic results. 5 COMMON DISEASES AND THEIR TREATMENT A quick algorithm for determining a possible diagnosis based on the information collected and the most common diseases in passerines is presented in Table 3 Additional diagnostic test and the differential diagnosis are listed in Table 4. The owner should be instructed to apply the ointment very sparingly or put a collar on the bird. and even these can cause significant haemorrhage if not used with caution. This results in very irregular or even a lack of effective blood levels. however. Water medication can be successful against mild infections of the gastrointestinal tract in which the drug may have a local effect in the gut. Antibiotics must be administered for at least 7 days and bactericidal antibiotics are preferred above bacteriostatics. the drug should be administered at the same time in both drinking water and the soft or egg food. there will with oral administration through the food and water be no measurable blood levels during the night in most cases. which can be the cause of disappointing therapeutic results and the development of resistance. resulting in self-medication several times a day (Table 2). In flock-treatment of small passerines. It is possible to increase water and food intake during the night by changing to a 24-hour light regimen. will disturb breeding and can induce moulting. A few drops swallowed during cleaning its feathers are sufficient to kill the bird. have an extremely high basal metabolic rate (K=129). The resulting effective blood levels after administration of drugs are often of a very short duration. The normal water intake is already irregularly spread over the day. To minimize risk. They obtain most of their daily fluid requirements from fruit. 413 . E. Klebsiella spp. It may be difficult. when they get the wrong food or no food at all (e. Overcrowding leads to aggression. coli. The breeding results are also disappointing in birds with an unbalanced diet. are often seen in mixed aviaries and individual pet finches. but it is not enteritis in the true sense. piscivorous. if the owner was away and someone else fed the birds). Haemochromatosis or iron storage disease is the most common non-infectious disease in soft bills including mynahs.1 Problems in mixed aviaries Nutritional problems especially those resulting from an unbalanced diet. Diets with 100 ppm iron or less have been recommended to reduce dietary 414 . and granivorous birds.g. A good starting point is a controlled feeding of three parts of a seed mix supplemented with one part soft food.5. Even granivorous birds need a certain amount of “soft bill food” as an unbalanced diet enfeebles these birds and can lead to problems with Enterobacteriaceae (eg. fructivorous. and omnivorous birds accumulate more iron in their liver than carnivorous. however. Trauma. Aggression is the main drive when aviaries are overcrowded or nesting sites or territory is involved.) and yeast infections (esp. Sick birds may attract aggressive behaviour. Often is diagnosed at necropsy. perhaps because they were too ill to eat (e. Hemorrhagic enteritis. because of an infection or intoxication) or. and insufficient nesting sites results in poor breeding results. This condition is often falsely called renal gout. and weakness are seen with hepatic haemochromatosis. to make the birds eat the soft food. therefore. when treating disease problems in these birds. This can range from a few feathers lost on the back of the head to cannibalism. more often. It should be differentiated from visceral gout based on impaired renal function or a high protein diet. insectivorous. white kidneys which are the result of uric acid precipitation in the collection tubules that occurs when birds do not drink. however. It is seen in small birds that for some reason did not eat at all for over 24 hours. but it should not be interpreted as nephritis or gout. The primary cause of many problems in Australian and other tropical finches is an unbalanced diet. Zebra finches are prone to indulging in this behaviour. Management and hygiene-related problems. In general. Candida albicans). Picking is a common problem in aviaries. improvement of the diet has to be the first objective. A similar interpretation should be given to swollen. It should be considered as a hemorrhagic diathesis (blood leaking into the gut).and endoparasites is a matter of constant attention. even within the same order.g. abdominal distension (hydrops ascites). Clinically dyspnoea. A typical sign is an empty crop. therefore the attacked bird should be separated and the underlying problem addressed. are management and hygiene-related problems due to location food and water utensils. The control of ecto. Many problems in aviaries. and Enterobacter spp. It can also be the result of inappropriate sexual behaviour of one or more dominant male birds. Articular gout is a poorly understood chronic condition with no relation to renal function. although and are exceptions and can infect any species. because most birds shown to have the disease had been consuming diets in excess of 100 ppm iron. The disease causes both young nestling mortality and a more chronic disease in which poor development and beak abnormalities predominate. zebra finches. In birds that are prone to develop iron storage in the liver. Herpesvirus and Cytomegalovirus. African silverbills [Lonchura malabarica cantans]. phlebotomy. Papovavirus and polyoma-like virus infections occur in finch aviaries across Australia and the United States and are probably more common than the number of cases actually diagnosed would indicate. 415 .sources of iron. A severe pancreatitis is found on histological examination. Although is often diagnosed in finches. These coccidia are said to belong to the Isospora lacazei group. respiratory dyspnoea and high mortality. a liver biopsy confirms haemochromatosis. Typical findings are skin-pox lesions. New information shows good effects of adding tannins or phytines to the diet. The diagnosis is based on the symptoms and can be confirmed by serology and virus isolation. In masked bull finches (Pyrrhula erythaca) a poxvirus has been demonstrated causing tumour-like lesions in the head-region and inside the beak.2 Infectious diseases Many infectious diseases are species specific. avian pox as a septicaemic problem is seen almost exclusively in canaries and other Serinus sp. this observation may prove to be more in line with what is available to feed birds than what is needed to prevent excessive iron storage. Because in a practical situation diets with less than 100 ppm iron are difficult to formulate. and Gouldian finches) with torticollis. Papovavirus. diets below 50 to 60 ppm can prevent induce an iron storage. In passerine birds kept in captivity. PMV-3 infections are commonly seen in many finches (e. One effective treatment.1 Viral Diseases Avian pox. SC) combined with a low iron diet (65 ppm) for periods as long as 4 months until the iron content in the liver of a toucan normalized.g. 5. Even diets with 100 ppm iron are in excess of the requirements for growth of poultry. A less invasive treatment is documented using deferoxamine (100 mg/kg q24h. The diagnosis is made by a specific fluorescent antibody test on liver and spleen impression smears and electron microscopic examination of the intranuclear inclusions Paramyxovirus. most birds appear to have their own coccidian species. Preventive vaccination is possible by the wing web method preferable in early summer. The necropsy is a specific. 5. which generally require 60 to 80 ppm. Radiographs revealing enlarged liver and ascites make the diagnosis. is usually performed in conjunction with low iron diets.2. These viruses cause respiratory problems in Australian finches. These are secondary pathogens. dermatitis. Campylobacter fetus subsp. and Staphylococcus spp. identical to pseudotuberculosis. and should be considered as a symptom of poor health or hygienic conditions. atoxoplasmosis or coccidiosis). Possible causes are unbalanced diet. Yersiniosis (pseudotuberculosis): An infection with Yersinia pseudotuberculosis is regularly seen in the wintertime in Europe in canaries and wild finches. and death. yellow. sinusitis. In fledglings. the clinician must search for the real cause. Treatment of choice is amoxicillin via the drinking water and the soft food. Treatment with an antibiotic does improve this condition but only temporarily. Salmonellosis (paratyphoid. jejuni is found very often in tropical finches. as well as. Society finches often are carriers without conspicuous clinical symptoms. Infections with Influenza virus have been reported in finches as well as imported mynahs 5. Carriers are not known in canaries. The antibiotics mostly effective are trimethoprim (with or without sulpha). extra water. Cocci infection. Enterobacteriaceae are regularly cultured from cases with diarrhoea in passerine nestlings (“sweating disease”). At necropsy a dark. In general there is a malaise with some birds showing conjunctivitis and rhinitis. coli (and other Enterobacteriaceae): These bacteria are isolated very often from the faeces or from the intestinal contents of diseased passerine birds.2. The antibiotics of choice are neomycin or spectinomycin because they are effective and not absorbed from the gut. like green food. are seen often. and cultivating the bacteria on special microaerophilic media confirm the diagnosis. or enrofloxacin. and at necropsy. The drug is administered via the soft food. Cocci will be seen in the impression smears. Demonstrating the curved rods in stained smears from the droppings or gut contents. “bumblefoot” and conjunctivitis. Local and systemic treatment with ampicillin or amoxycillin is the therapy of choice. Other primary diseases may be present (e. The clinical signs are not specific. There is no evidence for zoonotic importance of this infection. a few birds may die. will prevent dehydration. housing problems or management problems. with and without diarrhoea. The diagnosis is confirmed after culturing the microorganisms.2 Bacterial Infections E. pneumonia. amoxicillin. The clinical signs are abscesses. 416 . however. focal bacterial granulomas are found. The diagnosis is confirmed after culturing the microorganisms. The therapy needs to be combined with hygienic measurements.Other virus infections.g. Infections with Streptococcus spp. less often. although a chronic course is seen more often in salmonellosis. arthritis. especially in Estrildidae.) Infection with Salmonella typhimurium is clinically. swollen congested liver and spleen with small. On histological examination. slightly swollen liver. It is never seen as a flock problem. Mostly it are infections with atypical Mycobacterium avium or Mycobacterium-avium-intracellulare-complex and M. 417 . however. Cytology stain or cultures of the crop or cloaca should be performed. The classic tuberculosis with tubercles in the organs is seldom seen in small passerines. Avian tuberculosis. but are restricted to individual birds. pneumonia and air sac infections in canaries. 5. This is relatively uncommon problem in passerines. Mycoplasma spp have been isolated from canaries and many cases of conjunctivitis and upper respiratory disease in canaries respond to tylosin. toxoplasmosis and trichomoniasis. Listeria monocytogenes and Pasteurella multocida (cat-bite?).4 Protozoal Infections The most important protozoal infections in canaries are atoxoplasmosis. predominantly found on the mucosal surface and in the ducts of the glands. voice changes and dyspnoea. has been associated with chronic tracheitis.Enterococcus fecalis. In canaries infection with the yeast Macrorhabdus ornithogaster (formerly called Megabacteria)of the proventriculus is very common.3 Mycotic Infections Mycotic infections are not a significant problem in canaries.2. A polluted flower-spraying syringe. Ornithosis. The following bacteria are occasionally isolated from diseased passerines: Erysipelothrix rhusiopathia. Coccidiosis. No alterations are apparent at necropsy. macrophages loaded with acid-fast bacilli can be found in many organs. coccidiosis. genavense). can cause a severe necropurulent pneumonia and aerosacculitis. It is not uncommon to identify the yeast Candida albicans in cultures of the gastrointestinal tract of toucans and other soft bills. Especially cases of candidiasis are seen. there has been no conclusive work proving that Mycoplasma is associated with this syndrome. cochlosomosis and trichomoniasis are very common in finches. 5. Chicks that have poor daily weight gain or poor feeding response should be examined for potential bacterial or yeast overgrowth. except perhaps a dark.2. Pseudomonas infections. Other bacterial infections. but are much more common in tropical finches and mynahs. Inexpertly prepared sprouted or germinated seeds and dirty drinking vessels or baths can be the source of Pseudomonas spp. Atoxoplasma like infections and cryptosporidiosis are found only occasionally in finches. Individual infections with acid-fast bacilli are seen relatively often. infection causing a foul smelling diarrhoea. used for spraying the birds. Clinically affected birds have harsh respiratory sounds. Trichomoniasis. greenfinch and bullfinch). Cochlosoma sp. or infection of mice confirms the diagnosis. The diagnosis is based on demonstrating the flagellates in fresh and body-warm faeces from the finches. can cause many deaths among Australian finches fostered by these carriers. dark red coloured spleen. At necropsy trichomoniasis presents a thickened. diarrhoea. a huge. No effective treatment is known. 5. In the imprints of the liver. and death. living in the intestinal tract of Society finches. Acuaria skrjabini infections of the gizzard with mucosal necrosis were reported in adult finches in Australia Tapeworminfestations in insectivorous finches (particularly parrot finches and diamond firetails) are common and are regularly seen in Sturdidae. Serology. Cochlosomosis. opaque crop wall. The diagnosis can be made in a live bird with a crop swab. referred to by fanciers as “thick liver disease”. Infections with Trichomonas sp are sporadically seen in canaries. In wet preparations from the droppings large amounts of oocysts can be seen. sometimes spotted liver. and often an oedematous duodenum with vascularisation are seen. The clinical symptoms are huddling and ruffling of the feathers. The clinical symptoms are diarrhoea and emaciation. in the acute phase with necrosis. Atoxoplasma-like infections are seen in tropical finches. This infection is also a common problem in other European finches kept in captivity (e.Atoxoplasmosis is caused by Isospora serini in canaries. Canaries become blind many weeks after the infection. At necropsy an enlarged. occasionally neurological signs (20%). debilitation.2. Plasmodium and other blood parasite have incidentally been reported in passerines. In outdoor aviaries an infection with gapeworms (Syngamus trachea) is incidentally seen. Coccidiosis can be a problem in passerines of all ages older then 2 months. An enlarged liver can be seen as a blue spot at the right side of the abdomen caudal to the sternum. siskin. Therapy consists of strict hygienic measurements and treatment with coccidiostatic drugs.. Toxoplasmosis: In the acute phase of toxoplasmosis the birds (canaries and mynahs) may show severe respiratory signs. The flagellate. In cytology from the lungs trophozoites are easily demonstrated and in histology slides from the brains (pseudo)cysts can be found.g. mynahs and other Sturdidae. but common in Australian finches.5 Helminthic parasitism Worms are of no significance in small passerines. 418 . immunofluorescence on brain tissue slides. goldfinch. the spleen and the lungs the parasites are found in the cytoplasm of the monocytes. It is easily found and recognised in scrapings from the altered beak. Local treatment with any oil or 0. they are seen mostly Australian finches. KS. and Neocheyletiella media) and feather mites (eg. Epidermotidae.6 Arthropods Ectoparasites.5-4. Dermation spp) in the calamus of the feathers.6 200-400 2. including blood-sucking mites (Dermanyssus gallinae and Ornithonyssus sylviarum). which is caused by a papillomavirus. This infestation should not be confused with the so-called “tassel-foot” in the European goldfinch (Carduelis carduelis). Some selected normal haematologic and serum biochemical values in passerines (adapted from CARPENTER JW.5.5 Finch 45-62 2.5-4. DORRESTEIN GM and QUESENBERRY K.2.1% ivermectin applied locally will cure the birds.0 6-11 25-65 20-60 0-3 0-3 0-5 130-350 (600-1000) 9-13 190-350 2.4-4.6 3-8 20-65 20-65 0-1 0-1 0-5 150-350 200-450 3-5 Mynah 44-55 2. Table 1. 1996. Air sac mites.5 WBC (106/µl) Heterophils (%) Lymphocytes (%) Monocytes (%) Eosinophils (%) Basophils (%) Chemistries: AST (IU/L) LDH (IU/L) Ca (mg/dl) P (mg/dl) Glucose (mg/dl) TP (g/dl) 419 . Sternostoma tracheacolum are found occasionally in canaries. pp 156 and ALTMAN RB. skin mites (eg. RUPIPER DJ: Exotic Animal Formulary.6-5. p 1021) Measurement Haematology: PCV (%) RBC (10 /µl) 6 Canary 45-60 2. Saunders Co.5 4-9 20-50 40-75 0-1 0-1 0-5 45-345 120-350 5.3-4. Greystone Publications. Manhattan. 1997.4 1. Backericheyla sp. Knemidocoptes pilae is occasionally seen on the beak base of finches.0-4. CLUBB SL. MASHIMA TY. Meal mites (Tyroglyphus farinae) can cause unrest and irritation.1-13. 3000 200 . in soft food (mg/kg) 300 .1500 100 250 200 125 100-200 800-10003 100-200 200 100 80-100 100 000 IU 50 000 IU 400 200-400 200-400 150-300 150 50-100 250-400 Conc. one drop of 1% solution. 4 For the treatment of Candida albicans for 3-6 weeks. 21 days. Dosage regimens for chemotherapeutics and antibiotics for canaries and small passerines Drug Amoxycillin Ampicillin Chloramphenicol Chlortetracycline Dimetridazole Doxycycline1 Enrofloxacin2 Erythromycin Furazolidone Ivermectin3 Lincospectin Ketoconazole Metronidazole Neomycin Nystatin 4 1 Conc.300 1500 1000 200 200 200 200 200 100 100 200 000 IU 50 000 IU 400 400 400 100 400 Polymyxin Ronidazol Spectinomycin Spiramycin Sulphachlorpyrazin Sulphadimidine Trim/sulpha Tylosin 5 -----------------------------------------------------------------------------------------------------1 In case of ornithosis. 2 In case of ornithosis. alternative by topical application. 5 This dosage is for the trimethoprim part alone.Table 2. 30 days. 420 .500 2000 . 3 Concentration in MICROGRAMS/L.2000 100-150 1000 .400 1000 . in drinking water (mg/l) 200 . regurgitation. the infection spreads quickly.Juvenile.Canary . sometimes ATOXOPLASMOSIS neurological signs (20%) and death.Table 3. respiratory symptoms.Nestling . -Apathy. and there is much mortality between 1 and 3 days of age. -Very pale membranes visible by opening their beaks and weak in stretching their necks. TOXOPLASMOSIS BLOOD SUCKING MITES STERNOSTOMOSIS TRICHOMONIASIS -Severe respiratory signs. on eyelids. debilitation with scabs and pox-lesions. wheezing. sneezing. under one year of age . -Minor to severe respiratory symptoms with anaemia and sometime a high mortality. A low mortality. Females B L O O D . debilitation. head shaking and gasping. .Interior of the nests are yellow stained by diarrhoea of the nestlings.Loss of voice. The main complaint from the owner is usually a general depression in the bird.Mixed aviary Age: . 6 7 5 -Respiratory distress -Respiratory symptoms not main sign 6 -Dyspnoea.Any age 3 . MITES 421 4 -The youngsters show huddling and ruffling of the feathers. but seldom diarrhoea. Birds of all ages can be AVIAN POX affected and the mortality lays between 20 and 100%. respiratory distress. Mortality can be as high as 80%. the E. Diphtheric lesions can be found in the mouth and larynx. coughing. Diagnostic table for canaries and finches 1 Species: 2 12 7 3 4 5 2 .S U C K I N G can be found death sitting on the eggs. esp. which often results in blind birds after 3 months due to a panophthalmia. general illness and central nervous symptoms and iridocyclitis. blowing bubbles and emaciation. the feathers sticky. nasal discharge. . diarrhoea. commissure of the beak and in feather follicles. squeaking. COLI DIARRHEA youngsters stunt. decline of physical condition.Australian finch . 7 . clinically indistinguishable from pseudotuberculosis. diarrhoea.Nestlings and fledglings under the age of 3 months . nasal exudates and conjunctivitis. Weakness is often interpreted as a CNS-symptom. 13 16 14 15 12 .staining of the fledglings. SALMONELLOSIS -Especially in outdoor aviaries. often a obvious salivation and dyspnoea or diarrhoea in apathic birds CHLAMYDIOSIS MACRORHABDUS ORNITHOGASTER TOXICOSIS -Often after a weekend when some other than the owner fed the birds. The clinical signs are apathy.Birds grow thinner. more often chronic -Apathy.Sudden death of several birds 10 -Most infections are seen in winter. Sometimes black-stained STARVATION droppings or diarrhoea. and high mortality.Obvious wasting . but only low mortality 11 -Not specific.Age: .Diarrhoea not specific 8 -A general decline of the physical condition. diarrhoea. respiratory symptoms.Diarrhoea 9 8 . shrivelling and yellow. COCCIDIOSIS diarrhoea and emaciation. 422 . The mortality is low. . decline in food and water intake.Natural breed or foster parents 14 -From the age of 10 days until 6 weeks. -Several birds demonstrate a general malaise. Some birds may die. ruffling of the feathers. The foster parents COCHLOSOMOSIS show only watery droppings. huddling and ruffling of the feathers. debilitation. there are debilitation. difficulties with moulting and parts of or whole seeds in the droppings. CNS-symptoms. PSEUDOTUBERCULOSIS debilitation.All ages affected 13 . emaciation. 10 11 9 .Bengalese or Society finches as foster parents . The mortality is usually less then 10%. debilitation. with or without diarrhoea and some birds showing COLIBACILLOSIS conjunctivitis and rhinitis. STERNOSTOMOSIS head shaking and gasping. adult Estrildidae can show apathy and yellow diarrhoea or yellow solid CAMPYLOBACTER droppings due to large amounts of undigested amylum. The mortality is low. -Apathy. regurgitation. blowing bubbles and emaciation. 17 18 16 .Respiratory distress not the main symptom 17 -Respiratory distress. nasal discharge. -In nestlings the crop is bloating.Sudden death of several birds . 19 7 PARAMYXOVIRUS 11 18 . wheezing. In weanlings and CANDIDIASIS adult birds diarrhoea and moulting problems are more prominent. loss of voice. respiratory symptoms.Respiratory distress . .15 -High losses of nestlings. As long as these birds can still eat. coughing. mortality is low. sneezing.CNS-symptoms not a main symptom 19 -Torticollis is the main symptom. sometimes TRICHOMONIASIS diarrhea. and a thickened crop wall is relatively common.CNS-symptoms 423 . squeaking. Table 4. IFT or Elisa. Not important in small passerines. Exotic Animal and Wildlife Section. Culture Necropsy and demonstration of the agent by staining.m. Dorrestein. organ-smears or histological Toxoplasmosis AUTHORS ADDRESS G. Helminthic infestation Syngamus very occasionally. Necropsy Serology and demonstration of the parasite in brain-smears.nl 424 . Pet Avian. Detailed case history. when the toxin is not known. Parasitical examination of droppings collected between Coccidiosis 2 and 6 p. Mites Macrorhabdus sp Paramyxovirus Pseudotuberculose Salmonellosis Starvation Sweating Disease Sternostomose Trichomoniasis Demonstration of mites in the nest or bird-room crevities Demonstration of the yeasts in a scraping of the mucosa of proventriculus Serological and virologic screening. Haemorrhagic diathesis (bleeding into the gut) at necropsy Demonstration and isolation of bacteria in the faces.uu. The Netherlands Email: g. In the histology a pancreatitis Necrotic foci at necropsy in liver and spleen and agent isolation. histology and virus isolation In smears from the dropping the microorganisms can be demonstrated after staining with Diff-Quick. Utrecht University.m. Analysis of the situation for other factors in combination Colibacillose with the isolation. M. A direct wet preparation and/or a stained smear. Diagnostic necropsy and demonstration of the parasite Demonstration of flagellates in crop-swab. DVM.dorrestein@vet. Flagellates in a wet mount of fresh and body-warm faces Cochlosomose from the finches. A direct conformation often Intoxication impossible. Special hints for further diagnostics A necropsy and demonstration of the parasites in imprints Atoxoplasmosis of several organs Avipox Campylobacter Candidiasis Chlamydiosis Necropsy. Necrotic foci at necropsy in liver and spleen and agent isolation. Cultivation only on special media. PhD Department of Pathobiology. These have implications for the day to day medical and surgical care of birds and include. air sacs. development of the more heavily calcified avian egg and parent incubation. for example. An understanding of how birds evolved from reptilian ancestors is helpful in understanding the anatomy.B. The Cretaceous-Tertiary boundary extinctions of c. presence of pneumatic bones and fusion and strengthening of bones throughout the body. M.V.V. It is not possible for veterinarians to learn detailed anatomy of each of these species and clinicians are often challenged by types of birds they have rarely or never previously encountered. (Bird Medicine) KEYWORDS Bird clinical anatomy – Evolution . physiology and diseases of present day avian patients. respiration and renal function reflect birds’ reptilian ancestry as well as constraints of powered flight.Highbury Veterinary Clinic..000 species of birds on the planet.Therapoda ABSTRACT Birds are flying dinosaurs that trace their ancestry to bipedal therapods of the Mesozoic Era. Australia AVIAN CLINICAL ANATOMY FROM AN EVOLUTIONARY PERSPECTIVE P. Avian anatomy and physiology. (Hons). recovered 425 . including musculo-skeletal structure. 33 million years ago were key events in the emergence of modern volant families of birds. M.. reproduction. F. Macwhirter B. small clutch sizes compared with reptiles. Victoria. in Europe have been known to science since the 1860s but discoveries of the late twentieth century. more than any other group of vertebrates.A. a primitive bird from the Upper Jurassic.Sc. about 150 million years ago (mya). Burwood. nesting behaviour. lack of a diaphragm and use of air capillaries rather than alveoli.A. 1 INTRODUCTION There are currently over 9. 65 million years ago and the development of strong winds following the severing of the link between greater Australia and Antarctica c. unique jaw and beak structure. particularly that of Protoavis.Sc.A.C. Fossil remains of Archaeopteryx. 350 mya. When a wave of global extinctions occurred 245 mya. * Examples showing clinical relevance of avian evolution are highlighted with this symbol in these notes. Protoavis had pectoral girdle suggesting it was capable of limited powered flight while the pectoral girdle structure of Archaeopteryx suggests that this bird was primarily a glider. In the Carboniferous Period. lizards and snakes as well as the archosaurs which in the warm humid climate of the Mesozoic Era gave rise to the crocodile family. the dinosaurs and. The earliest amniotes were anapsids without any lateral openings on the side of the cranium posterior to the orbit. gave oxygen in exchange for carbon dioxide. In this context the amniotes emerged. Birds subsequently evolved avidiapsid crania in which the two lateral cranial openings merged into a single opening. 2 THE AMNIOTE EGG Since multicellular organisms first emerged on earth some 4 billion years ago. 426 . one above the other. birds. enabling a vastly increased pool of natural variation and selection pressures that favoured organisms able to reproduce independently of an aquatic environment. Independently from primitive anapsids the synapsids (mammals) evolved with a single lower opening on the skull posterior to the orbit and the diapsids (reptiles) evolved with two lateral openings. These were vertebrates that produced eggs containing specialised membranes that provided the developing embryo with a liquid environment. Pangaea. large amphibians over most of Pangaea were wiped out while descendants of the small reptiles survived and evolved to fill a wide range of ecological niches. These openings allowed for larger. retained yolk sacs. more powerful jaw muscles for chewing and capturing prey. jaw function and injury repair. periods of gradual diversification of life forms have been punctuated with periods of abrupt extinctions (DE DUVE 1995). Flora and fauna between the two landmasses intermingled. moved south and coalesced with the southern continent of Gondwana to become a single land mass. Tortoises and turtles have been traditionally classified as members of the Anapsida. These included the progenitors of turtles. a sliding action of the quadrate bone that enabled independent movement of the upper beak. later. stored food as yolk and isolated nitrogenous waste. which included Europe and parts of North America had emerged from the sea. the pterosaurs. Interestingly. lycopods and horsetails. * Clinical relevance: hatching difficulties.from Triassic (225 mya) fossil beds in Texas and a trove of feathered dinosaurs from the Cretaceous (about 125 mya) from China have changed and expanded current knowledge of the evolution of birds. allowing for streptostyly. The feathered dinosaurs of China most likely did not fly but used their feathers for other purposes. crocodile shaped amphibians called labyrinthodonts dominated a land vegetated with primitive psilophytes. the much older bird. The continent of Laurasia. respiration. The paucity of the avian fossil record. 4 HOW DID REPTILES EVOLVE INTO FLIGHTED BIRDS? If a species is to survive and multiply. flighted and non-flighted avian species of today has been the subject of much speculation. Bird bones are particularly fragile and do not preserve well. buoyancy or flight 427 . evolutionary change needs to confer immediate advantage for the next generation. 65 mya. Efficient flapping flight appears to have occurred later in the sequence. Feathers are the key difference between birds and (other) reptiles but the earliest birds did not fly and the evolutionary steps between ectothermic reptiles and the wide diversity of endothermic. the expansion of winged insects and emergence of flowering plants c. ‘la grande coupure’ . Bipedalism with feathers for thermoregulation. reproduction. canids and humans. global cooling and increased winds. possibly due to a meteor crash near present day Mexico c. Critical bio-geographical events over the 225 million year period since the first appearance in the fossil record of possibly feathered reptiles have included • • • • • • • the gradual break up of Pangaea and Gondwana due to tectonic plate movement. phylogenetic analysis and analysis of fossils with possible pro-avian features emerging from both Asia and North America have lent weight to the case that birds are embedded in the bipedal dinosaur class of therapoda at the highest node of dinosaur evolution. fluctuating climates and the emergence and global expansion of mammalian species. of the dinosaur family (CHATTERJEE 1997. felids. indeed the only the only living members. They are living members. ongoing sea level changes.3 WHEN AND WHERE DID FEATHERED REPTILES FIRST APPEAR? It has been strongly argued that birds should be placed as a subgroup within Reptilia rather than given equal rank with Mammalia and Reptilia. However. 120 mya the Cretaceous-Tertiary (‘K-T’) Boundary Disaster that marked the end of the Mesozoic Era. Crocodiles are their closest living relatives. renal function and musculo-skeletal structure. particularly from the Mesozoic Era. leaves many blanks in the sequence but key events in the transition from reptiles to birds must have included the development of feathers along with changes in thermoregulation.the separation of South America and then greater Australia from Antarctica with resultant circumpolar currents. 33 mya. c. especially pinniped. nesting behaviour. so it is not surprising there are gaps in the fossil record. MARTIN 2002). may have been the key initial benefits that feathers provided (PRUM and BRUSH 2004). still with reptilian teeth. In common with other therapods. most birds have the phalangeal formula of 2-3-4-5 with absence of the fifth digit. tarsometatarsus and intertarsal joint structure and disease. scaly and feather mites.In general. ectotherms are at an advantage in hot climates where food sources scarce but endotherms perform better when the climate is cooler and food sources abundant. using a long neck and bill. * Clinical relevance: tibiotarsus. birds that first appear in the fossil record around 120 mya showed features of a reduced tail. Both features improving their manoeuvrability and capacity for powered flight. around the shores of the Tethys Sea where Archeopteryx lived or in the Triassic swamps of Texas where Protoavis lived. loon-like Cretaceous marine bird from the northern hemisphere. leg fractures & repair. pododermatitis in webbed and other feet. Later in the fossil record. in addition. balance and perhaps buoyancy. Evolving near the Antarctic circle. Feathers do not appear to be modified scales but rather emerged in therapod dinosaurs as independent tubular structures that became progressively more complex. these birds might have 428 . a toothed. Those living at a distance. walked or ran on two legs rather than perched on small branches as their more curved footed descendants came to do. It is likely that birds and bird-like reptiles living near the impact would have had little chance of surviving the initial explosion. couldn’t fly but feathers aided in buoyancy and it used its large feet for swimming. Enanatiornithines. The flat foot structure of early fossil birds suggest they leaped. They perhaps leaped or hang-glided. Probably in response to a bipedal cursorial lifestyle proximal tarsal bones fused with the tibia to form the tibiotarsus and distal tarsal and metatarsal bones form a single tarsometatarsal bone. 5 TERTIARY BIRDS. Such insects. pygostyle injuries in wing clipped birds. Insulation. * Clinical relevance: problems associated with large avian eyes. for example southern South America. to catch prey. it would seem that a variety of primitive birds were widespread at the time of the great Yucatan meteor crash of 65 mya. a deep keel bone and a pectoral girdle structure with a rudimentary triosseous foramen suggesting the bird was capable of limited powered flight. fossil evidence suggests that Protoavis also had stereoscopic vision. a pygostyle and fused carpal and metacarpal bones. their forelimbs free for balance and other purposes. rather than flight. signs of pituitary tumours. THE K-T BOUNDARY DISASTER Given the global distribution of enantiornithines and more than a dozen other genera of land and sea based Mesozoic bird fossils. fires and the ensuing long dark winter. stress lines and feather formation. waded. Antarctica and Australia (which at that time were still conjoined as Gondwana) may have escaped the worst of the global disaster. Both species had feathers and were almost certainly endothermic but. Winged insects pre-dated the emergence of birds. along with aquatic creatures could have provided an abundant food source. Hesperornis. penguins. dark conditions and could provide bird populations from which the northern hemisphere could be repopulated. Grebes. * Clinical relevance: Megapod reproduction. By producing a more heavily calcified. less porous egg shell in a shell gland. For both birds and reptiles an excretory system based on the production of insoluble urates (rather than soluble urea) enabled waste material to be compartmentalised within the egg but away from the developing embryo in a nontoxic form and the capacity to adapt to dry conditions. * Clinical relevance: Ratite. Fossil evidence and distribution of endemic bird species suggest that following major extinctions at the K-T boundary. For pro-avians. cloacoliths. The ability to fly or swim would have aided dispersal. particularly in cool climates. gout. partially endothermic avian ancestors may have nested in burrows as. Kidneys became compressed into fossa in the synsacrum. The ability of feathered pro-avians to use their own body heat to speed up the incubation process may have been advantageous in the short term in enabling birds to make use of nest sites in tree hollows. bird eggs became less prone to desiccation and birds could exploit nesting sites above the ground. Bony-toothed pelicaniformes were found globally (WARHEIT and KENNETH 2001). Alternatively. Urogenital adaptation and nesting behaviour Birds’ reptilian ancestors had two ovaries and laid large clutches of eggs simultaneously which were incubated in nests on the ground in warm. penguin and pelican affinities and diseases. using body heat to shorten the incubation time (and later developing specialised brood patches) would have been an advantage in reducing risk of predation and enhancing survival of their young. some penguins (another ancient avian family) do today (KAVANAU 1987). penguin-like birds using their wings as flippers and loon-like marine birds using their feet as paddles were amongst primitive avifauna of the southern regions of Early Tertiary times. an ancient family that builds floating nests. Crocodiles. incubation patterns in waterbirds. still nest in this manner. It would have also favoured birds having small enough clutches to incubate against the parent’s bodies. branches or bushes but this also required a change in egg structure to enable the embryos to survive in conditions with lower humidity. * Clinical relevance: commorant and cockatiel posture. feathered. Laying small clutches of eggs individually rather than as a simultaneous clutch eliminated a need for two ovaries and pro-avian bodies could be streamlined to contain a single ovary and oviduct. The emergence of medullary bone lay down of calcium in egg laying females would have assisted this process. moist conditions and gave rise to precocial young. for example. 429 . Flight and the avian urogenital ‘package’ developed hand in hand. renal function. Flightless land birds. birds closest extant reptilian relations.been better adapted to the cool. might be a model for this type of incubation. Western Gondwana might have been the site of geographic origin of many bird families of today. kiwis. Fossils of giant penguin-like birds have been found near Adelaide. The Drake Passage between South America and Antarctica opened c. C. but they require strong muscles for the down beat – a function performed by the superficial pectoral muscles. In avian ancestors rotation of the coracoid to the front of the chest and the scapula to the flat of the back left the glenoid in a dorsolateral position. This caused global cooling and strengthened coastal and trade winds. so coined by the Swiss palaeontologist Hans Stehlin. altricial young could be produced from smaller eggs relative to the size of the adult bird.the great cut. Volant birds generally have dorsoventral rather than lateral flattening of the body as the centre of gravity needs to be below the extended wing rather than above it (SIMKISS 1963). temperatures dropped and global cold water currents carried rich marine food sources northwards (FLANNERY 2000). egg binding. South Australia date to these times (VICKERSRICH and HEWITH-RICH 1996). 33 MYA Following the K-T crisis. This positioning of the scapula facilitates climbing as well as the formation of the wing and it is also seen in mammalian climbers. narrow and fixed compared with the broad triangular scapula of climbing animals. 6 LA GRANDE COUPURE. The effect was magnified as polar ice caps expanded. This could have been an advantage for many flighted birds and might offset the greater parental care required to rear altricial young. egg yolk related peritonitis. By 40 mya. However in the avian model the scapula is strap-like. Flighted birds do not require strong muscles to raise their wings as ‘lift’ performs this function. ‘hernias’. Musculo-skeletal adaptations The strong circumpolar winds that followed la grande coupure may have been a key factor favouring birds with deep keel bones that were good at flapping flight. The two deep pectoral muscles (supracoracoids). around 33 mya. winds strengthened. 35 mya and finally. a long submarine rise that stretched from Australia to Antarctica severed allowing bottom water of the Antarctic circumpolar current to flood between the two continents for the first time. While primitive birds would likely have hatched precocial young. It was from this time that a dramatic expansion and diversification of flighted bird families occurred. Beginning 60 mya Australia began to unzip itself from Antarctica as tectonic plate movement slowly drew the continent northwards. another critical biogeographical event for evolving avifauna was la grande coupure . including primates. 430 . whose tendons pass through the triosseous foramen and insert on the humeri. India had crashed into southern Asia and the Himalayan Mountains were rising. help to produce lift by altering the angle of attack and adjusting curvature of the dorsal aerofoil surface of the wing through their indirect effect on the patagial membranes.* Clinical relevance: medullary bone lay down. both of which are cornerstones for leaping. spine and synsacrum.* Clinical relevance: distinguishing bones of the pectoral girdle on radiographs. replacement of teeth with lighter beaks fusion and strengthening of bones of the limbs. Protoavis and the Enantiornithines but the upsurge and explosion in diversity of birds capable of flapping flight occurred after 30 mya when pectoral girdle and deep sternal confirmation typical of modern carinates was refined and became widespread. shortening of the tail to become the pygostyle. Almost all other birds have well developed neopulmonic parabronchi. the neopulmonic parabronchial net. * Clinical relevance: patterns of bone fractures and coracoid and wing injuries. development of uncinate processes on the ribs. swivel carpal joints for wing folding enlargement of the ulna to which secondary wing feathers attach. Respiratory. both primitive avian families. Storks. Ratites and penguins. have paleopulmonic parabronchi in which air flow is caudo-cranial. digestive and immune system adaptations Respiratory systems of birds differ significantly from reptiles in their extensive development of the air sacs. fused clavicles forming the furcula. the enlargement of the coracoid. swimming or flight. presence of pneumatic bones and use of air capillaries rather than alveoli. Development of flight feathers with asymmetrical closed pennaceous vanes The seeds of some of these changes could be seen in Archeopteryx. cormorants and cranes have this system as well as an additional network of parabronchi. These features generally make for lighter bodyweight and efficient respiration. lack a diaphragm. spinal injuries at the flexible joint T6-7. development of the keeled sternum. running. Birds’ rigid skeletal systems appear to have functioned as a bellowslike apparatus in breathing and aided in streamlining and lightening birds’ bodies for swimming or gliding before birds took to the air in flapping flight. unidirectional and linked with air sacs. the triossesus foramen which enabled the tendon of the deep pectoral muscle to attach to the head of the humerus and control the angle of attack of the leading edge of the wing. in which air flow is bi-directional. In addition to the change in the centre of gravity and reduction and fixation of the scapula. 431 . effects of pectoral muscle injuries. sternal injuries associated with wing clipping. development of the alula from the second digit of the wing. skeletal refinements useful for flapping flight in birds include: • • • • • • • • • • • • • pneumatic bones. Bird flight. Although not reflected in the fossil record. London: Hutchinson Educational Ltd. behaviour and evolution. 2. The Rise of Birds. (1997). 72 .Avian digestive systems reflect diet diversity and weight constraints of powered flight. Which came first. BAIRD R. 1 . SIMKISS K. KAVANAU JL. the origin and evolution of life on Earth. use of abdominal breathing tubes for anaesthesia. 6. Los Angeles: Science Software Systems. New York: Basic Books 1995. 225 million Years of Evolution. will be used. 5. * Clinical relevance: distribution of (e. large caeca may be present in herbivorous ground dwelling birds such as gallinaceous birds but are reduced or absent in passerines. Vital dust. while all present day birds lack teeth.640. nature of allergic reactions in birds and response to intradermal skin testing. 7.) Aspergillosis lesions. 9. 1 – 8. 9 CITATION INDEX AND FURTHER READING 1. MARTIN L. 3. PRUM R and BRUSH A. IgA.g. In: VICKERSRICH P. The eternal frontier. 8. IgG and IgE as occur in mammals.224. cockatiels and budgerigars. The rise of birds. Baltimore: John Hopkins University Press 1997. For example. IgA and IgY. Melbourne: Text Publishing Company 2000. as seen in reptiles rather than IgM. 809 . 225 million years of evolution. with kind permission of the publisher John Hopkins University Press. the key avian immunoglobulins are IgM. An early archosaurian origin for birds. Melbourne: Monash University Publications 1996. 10. BAIRD RF and RICH TH (eds): Vertebrate palaeontology of Australasia. 8 ACKNOWLEDGEMENT Illustrations from CHATTERJEE S. DE DUVE C. Beijing: IOC Proceedings 2002. 7 CONCLUSION Discoveries being made in the fields of palaeontology and molecular biology are giving greater insight into avian anatomy and physiology from an evolutionary perspective. 1963. Avian fossils from the quaternary of Australia. FLANNERY T. 373 . Using an evolutionary framework is a useful aide in trying to understand and deliver quality veterinary care to our diverse range ‘flying dinosaur’ patients. MONOGHAN JM. CHATTERJEE S. 4. Birds as dinosaurs.82. Inc 1987. Lovebirds.46. in conducting this Master Class. Beijing: IOC Proceedings 2002. 13 . the feather or the bird? Scientific American 2004. 9.870. disease patterns of Coturnix and Turnix quail. SERENO P. 432 . M. 128 Highbury Road. Sydney: Reed 1993: Wildlife of Gondwana.V. (Hons).A.C. The seabird fossil record and the role of palaeontology in understanding seabird community structure. In: Schreiber E. 3125 Australia Email: patvet@bigpond.. Burwood. 1 .808. Victoria. 12. MONOGHAN JM. and Burger J.Sc. Boca Raton: CRC Press 2001.com 433 . 721 . BAIRD RF and RICH TH (ed): Vertebrate palaeontology of Australasia. 11. Birds.A. VICKERS-RICH P. 13. 17-56.A.V. Macwhirter B.Sc. Melbourne: Monash Univ Publications 1996. The Mesozoic and Tertiary history of birds on the Australian plate.. AUTHORS ADDRESS P. VICKERS-RICH P and HEWITT-RICH T.A. In: VICKERS-RICH P.148. M. WARHEIT and KENNETH. (Bird Medicine) Highbury Veterinary Clinic. Scientific American 1979.10.B. WILSON B. F. Biology of Marine Birds. University of Leipzig Germany IMAGING NON-INVASIVE DIAGNOSTIC TECHNIQUES OF THE UROGENITAL TRACT M. lethargy.E. diminished appetite up to emaciation. Prof DrMedVet.Ultrasound ABSTRACT Radiology and ultrasound are important diagnostic tools in birds with alterations of the urogenital system. which pass through the kidneys. endocrine and neoplastic origin can be diagnosed. DrMedVet KEYWORDS Urogenital system – Kidney – Testes . Krautwald-Junghanns. M. Dipl ECAMS. echogenicity). Both non-invasive techniques have its advantages and limitations. The combination of them gives information about the organs’ size and inner structure (radiodensity.Clinic for Birds and Reptiles. 434 . More specific are clinical findings like non-traumatic unilateral or bilateral paresis of the legs. dehydration. defecation problems and changes in the urinary output are common signs of renal disease (LUMEIJ 1994).2 Indications for non-invasive diagnostic techniques Clinical signs indicating renal disease are generally non-specific and frequently superimposed by secondary changes caused by renal dysfunction.Ovary – Oviduct – Imaging techniques – Radiology . Subcutaneous urate tophi or urate accumulations in the joints are signs of hyperuricemia caused by renal disorder. 1. Pees.1 Anatomy The paired kidneys are situated in the dorsocaudal part of the thoracoabdominal cavity. alterations of traumatic. 1 KIDNEYS 1. Dyspnoea. Since a urinary bladder and a urethra are missing. Besides infectious diseases. Each kidney is divided into three lobes all containing cortical and medullar tissue. caused by compression or inflammation of branches of the lumbosacral plexus. the urine is conducted via the ureters directly into the urodaeum of the cloaca. Active gonads must not be confused with kidney tumours. urate tophi in the kidneys are visible as radiodense shadows.g. Nevertheless blood chemical values within the physiological range do not exclude a renal disease.Alterations of blood chemical parameters like uric acid are further hints for renal disease. 1. However. the surrounding air sacs are displaced and it might be difficult to evaluate the cause of distension. crystalline deposits in the kidney is readily demonstrated in both of the standard planes. 435 . gout and vitamin A deficiency.3 Radiology Survey radiographs (laterolateral and ventrodorsal projection) provide information about the size and radiopacity of the kidneys. and/or cases of radiologically diagnosed soft tissue masses in the region of the kidneys (HOFBAUER and KRAUTWALD-JUNGHANNS 1999). Enlargement of the renal shadow can be assessed in the laterolateral projection. In these cases. Barium sulphate contrast of the gastrointestinal tract point out space-occupying lesions like renal tumours or cysts. They are also seen after prolonged fluid deprivations. In cases of massive enlargement of the kidneys. since surrounding air sacs provide a good negative contrast. However in some severe cases only. kidney cysts and neoplasia. it may be necessary to have recourse to contrast radiography in order to define the kidneys against the surrounding organs. Non-infectious causes are vitamin A deficiency. Contrast studies using organic iodine compounds for visualisation of the kidneys/ the ureters are possible and may provide information about the functional status of the urinary system. due to anatomical peculiarities in comparison to mammals their use is limited in birds (KRAUTWALD et al. 1992). They are generally non-specific signs for renal insufficiency of different aetiologies. They occur in chronic kidney infections indicating renal insufficiency. The latter is one of the most frequent aetiologies of renal disease in the budgerigar (Melopsittacus undulatus). only high-grade alterations can be diagnosed. there tends to be an increased density of the kidney shadow. 1. In the ventrodorsal projection. The presence of radiodense. Such crystals are evenly distributed throughout the organ and are due to the precipitation of uric acid and/ or renal calcinosis. chlamydiosis). abnormalities like crystalline inclusions can also be detected.3. Besides the size. but they are not specific for gout. but this is difficult to evaluate as the bony pelvis often obscures the image.1 Plain radiology Enlargement of the renal shadow is interpreted as a non-specific sign of many generalised infections (e. Furthermore indications for sonographic examination of the urogenital tract consist of any disorder which is more or less connected with abdominal distension. they are superimposed by the gastrointestinal tract. often in association with a high blood uric acid level. This could involve GI contrast or renal contrast. In severe dehydration. complete absence of a contrast image in the kidney or ureter occurs sometimes when renal tumours or cysts are present. in birds with organ enlargements. gonadal tumours). cloaca and terminal gut are visualised 2 – 5 minutes after the injection.80 % solution of organic iodine compound or a solution containing 300 – 400 mg of iodine per ml (800 mg/kg intravenously (NEWELL 2000). usually into the ulnar vein. In cases of kidney enlargement. Kidney tissue is demonstrated in the area between the intestines and the total reflexion of the spinal/pelvic bones. but also the density (echogenicity). abdominal swelling and especially in cases when ascites is present. heart and pulmonary arteries are demonstrated 10 seconds after the injection. where it is desirable to augment the demonstration of those organs. such as in sinography where it is injected directly into the sinus. or by some other route. unless this has already been established in the plain radiograph (enlargement of the kidneys.1. • In isolated cases: diseases of the other parenchymatous organs. Furthermore ultrasound is ideal for diagnosis of cystic alterations within kidney tissue (HOFBAUER and KRAUTWALD-JUNGHANNS 1999). • Conditions of the kidneys and ureters accompanied by decreased function as. kidneys and ureters are shown 30 – 60 seconds after injections. It may be necessary to aid emptying the gut by introducing liquid paraffin into the cloaca.2 Urography Renal contrast radiography is indicated in some instances to obtain information about functional disturbances of the urinary apparatus. The dose is 2 ml per kg body weight. For urography the organic iodine compounds is warmed to body temperature and injected slowly intravenously. A reduction or. 436 . if this could not be achieved by the plain radiograph or GI-contrast study (for instance in ovarian cysts. 1. rarely orally (an example for oral use is Gastrografin®). on the preparation used and on the iodine concentration of the medium. Aorta. For urography the patient should be fasted for about 2 hours before the contrast medium is given. Indications: • Diseases of the kidneys and ureters that are associated with alteration in their size or shape. In contrast. • Defining the kidney against the surrounding organs. Elimination of the contrast medium is retarded in renal insufficiency due to a variety of causes. for instance. not only the size of kidney can be assessed. the sonographical examination may provide more information than any other imaging technique. as a 70 % . The quality of the contrast obtained depends on the concentrating capacity of the kidneys.3. Organic iodine compounds are administrated intravenously. in renal neoplasia or cysts and in renal insufficiency from whatever cause when excretion is retarded. for instance. in cases of acute nephritis).4 Ultrasonography Ultrasonography is of limited use for the assessment of the normal-sized kidney. indeed. in cases of organ enlargement.1 Approaches for ultrasonography of the kidneys There are three possible approaches to the organs of the avian urogenital tract (HOFBAUER 1996): The cranioventral approach. both kidneys can be compared to each other. A direct comparison of both kidneys is not possible in this view. With this view. the major part of the kidney tissue can be examined for parenchymatous alterations (changes in echogenicity.4. the kidneys are presented lying in a W-shaped total reflexion caused by the pelvic/spinal bones. this technique is of little practice importance in pet birds. the kidneys can be examined in their long axis (longitudinal to the spinal column). see below). This is possible not only in large birds but also in smaller ones (e. the examination is possible and both the size and the parenchyma can be assessed.g. This led to better imaging. in the flank directly behind the last rib Due to anatomic dissimilarities these approaches are not available in all species.1. the gently caudally sloped scanner has to be angled about 70° to obtain a better image of the organs. by gentle pressure with the scanner onto the abdominal wall. within the depressions of the pelvis and the surrounding abdominal air sacs. With a cross section (the beam plane perpendicular to the spinal column). mid-abdominally directly behind the Margo caudalis of the sternum • The caudoventral approach. particularly with the lateral approach. This has been described in birds with a bodyweight of more than 2 kg (HILDEBRANDT et al.4. Kidney size: Whereas the ultrasonographic visualisation of the normal kidneys is difficult. it is advantageous to compress the air sacs. and also due to the rather small size along the ventrodorsal direction of the ultrasound waves. at the present time. This is due to its position along the vertebral column. and measurement of the size can be done. This should be done after identifying the renal tissue in the cross section. situated between the pubic bones of the pelvis • The lateral approach. With this view. located underneath the body wall. In this section view of the kidneys they are presented round to oval.2 Examination methods and evaluation criteria The sonographic demonstration of the normal kidney by transcutaneous ultrasonography is in most cases not possible. budgerigars) without problems. Using special sonographic equipment and transintestinal examination technique the demonstration of the normal kidney is possible. 1995). Kidney parenchyma: The echotexture of the enlarged inflamed kidneys is homogenous and rather • 437 . By turning the transducer about 90 degrees. but also in the ventral approaches. Furthermore. On account of the complications caused by the caudal air sacs. not only in the lateral. However. due to the high costs of equipment and the limited possibilities of application (small size of most avian patients). 1. an ultrasound-guided puncture of the fluid-filled cavities is easy to perform and the fluid can be examined. GENITAL TRACT 2. In the other cases the patient owners report nonspecific symptoms which are also applicable for other diseases (for example respiratory disease). changes in posture. The size of the gonads and the oviduct depends highly on sexual maturation and activity. However. the vertical cross-section is preferable since the comparison of both kidneys is possible with this view. In cases of renal cysts without renal enlargement. Behavioral changes of the patients are also frequently observed. Sometimes. palpation can even lead to false negative findings. the history often contains obvious indicators for a reproductive disorder (abnormal eggs. dyspnoea. multiple poorly echoic foci mark necrotic areas within the irregular masses of the tumours. In these cases. tail wagging.2 Indications for non-invasive diagnostic techniques Clinical signs in birds with disturbances of the genital system include abdominal swelling. blood chemistry) should be taken into account before making diagnosis. but the reproductive tract of the hen consists only of a left ovary and oviduct. cranioventrally to the cranial lobes of the kidneys. The testes are paired. often with marked distal acoustic enhancement.anechoic with no recognizable inner structure. 2. They appear sonographically as clearly defined. diagnosis of renal gout by means of ultrasound is difficult. Neoplasia is visible as voluminous and often rounded single masses. it might be difficult to differentiate between renal and ovarial cysts. multiple growths as well as cases that are complicated by peritonitis cannot be definitively diagnosed by palpation (BOWLES 2001). Uric acid depositions and/ or calcifications cause reflexions (increased echogenicity). furthermore. In these cases. frequently with diffuse inhomogenous echotexture. the renal tissue appears more inhomogeneous. palpation proved to be a simple and reliable diagnostic technique for eggs with calcified shells in the caudal abdomen. cases of 438 . chronic egg layer). the patient history is only indicative in about half of the cases. If the hen has already laid eggs. which is often significant higher compared to the normal renal tissue surrounding the neoplasia. Also colour Doppler ultrasonography is possible for detection of the blood flow within the tumour. Besides neoplasia. other techniques (radiology. also cysts of the kidneys are easy to detect by means of ultrasound. However. laminated and thinshelled eggs. endoscopy. For identifying single-sided tumorous alterations. rounded anechoic structures. ectopic eggs. In cases of egg-binding. defecation problems and rear limb paresis/ paralysis (JOYNER 1994). depression/ anorexia. Therefore the results of the clinical examination should be confined by means of radiology and/ or ultrasound. Indications for sonographic examination of the genital tract consist of any disorder which is more or less connected with abdominal distension. 2. However.1 Anatomy The gonads are situated in the dorsal part of the thoracoabdominal cavity. as they occur. Also in birds with ovarial cysts and gonadal tumours. size. In birds with egg retention the egg can often be palpated. By this means. provided it is calcified and not situated too deep in the thoracoabdominal cavity. apathy. Radiography is helpful in diagnosing shell alterations. Medullar bone is therefore a helpful finding in radiographs with otherwise unclear findings. neoplasia of the testes may lead to emaciation. Laminated or thin-shelled eggs may be the cause of egg-binding in birds. be helpful (MC MILLAN 1994). can be fairly easily ruled out. are often associated with other radiographic features such as polyostotic hyperostosis and abdominal hernia. number and position of eggs. for instance.radiologically diagnosed soft tissue masses in the urogenital region and/or formation of medullar bones. In males (particularly budgerigars). such as cystic changes of the ovaries and oviduct and gonadomas. A radiograph is not only necessary to provide information about the form. Radiographically diagnosed medullar bone and contrast radiographs of the gastro-intestinal tract can. Alterations in the size and shape of the gonads. 439 . dyspnoea. changes that occur in conjunction with an increased estrogen level. such as tumours of the liver and kidneys. However. cannot be ruled out. Oviduct In some cases of salpingitis the oviducts shadow is enlarged and more dense than normal. it is also found in other disease processes. Nevertheless. however. common differential diagnoses in budgerigars. 2. anorexia and signs of feminization (in budgerigars: discoloration of the cere from bluish to brownish). Gonads In respect of the gonads one must differentiate between pathological enlargement and physiological activity but the clinical findings give the necessary indications. it can also help to decide between their surgical and non-surgical removal. which have radiographically diagnosed medullar bone. it is not possible to differentiate radiographically between eggs without a calcified shell or with a very thin shell and other soft tissue masses (for example tumours) in the thoracoabdominal cavity. multiple and deep intraabdominal situated eggs and is therefore an indispensable tool in choosing a therapy. medullar bones are often seen in radiographic examination. and no definite conclusion on the existence of a pathological process can be made on this basis (KOSTKA 1992). in cystic ovaries.3 Radiography A definitive diagnosis of egg-binding is reached with the help of radiography in 30% of the cases. Dyspnoea may be caused by distension of the air sacs. Unfortunately. Budgerigars are particularly prone to gonadal neoplasia which causes massive enlargement of the organ resulting in pressure on the air sacs and ventrocaudal displacement of the gastrointestinal tract (seen with barium-sulphate contrast studies). rear limb paresis due to compression of the nervus ischiadicus. varying echogenicity as well as. but there may also be evidence of an abdominal hernia.4. often with diffuse inhomogenous 440 . Its advantage is the direct diagnosis of soft tissue structures. 2. 1995) Gonads Testes: The sonographic demonstration of testes is only successful in the case of highly sexually active birds. It is. should be considered in cases with nonspecific histories. By transcloacal sonography a sex determination in birds with more than 2 kg BW is possible (HILDEBRANDT et al.4 Ultrasonography Ultrasonographic examination proved to be a helpful tool in diagnosing genital tract physiology and abnormalities (HOFBAUER and KRAUTWALD-JUNGHANNS 1999). Both methods. hyperechoic string surrounding the parenchyma.4. The parenchyma of this organ shows a delicately granulated structure of average echogenicity.1 Approaches for ultrasonography of the genital tract These are the same as described for the examination of the kidneys (see 1. Tumours appear as voluminous and rounded single masses. possible to recognize developed follicles. although testicular vessels normally cannot be identified. in most cases. laminated and thin-shelled (non-calcified) eggs and neoplasia as well as pathological processes in the oviduct (salpingitis). On the radiograph the laminated eggs produce a distinct soft tissue shadow. as opposed to radiography. The closely fitting testicular serosa occurs as a smooth. inflammatory processes and other alterations in conjunction with enlargement of the organ are principally sonographically observable. for example. Sonographic examination again allows a clear characterization of cysts.2 Examination methods and evaluation criteria General comments: The demonstration of the healthy gonads by transcutaneous sonography is only successful in cases of highly sexually active birds. Radiographic and sonographic examination therefore compliments one another in cases of suspected genital disease. in which extensive contrast studies are necessary.4. displacement of the gastro-intestinal tract and an increased density of the long bones. Cystic degeneration of various organs. Laminated eggs are typified by their shape. also have their diagnostic limits. especially of the liver and the kidneys must be considered. This differential diagnosis can be ruled out with the help of a contrast study of the gastro-intestinal tract with barium sulfate and/ or a sonographic examination. For the evaluation of the shell quality of intact eggs. Neoplasia. a fluid perimeter.The eggs shell may be abnormal in salpingitis. 2. sonographic examination is only appropriate in combination with radiography. however. As differential diagnoses neoplasia of other internal organs.1). 2. however. especially kidneys and ovaries. since not all changes are visible. its reflexion and absorption of the sound waves prevents the examination of the inner structures of the completed egg. Beyond that. Furthermore. the quality of sonographic images depends on the phase of development and. However. In polycystic abnormalities this is evident in the parenchyma. In sonographic demonstration and interpretation both ovarial parenchyma and follicles of varying degrees of maturity were principally regarded as a morphological unit. Ovary: The sonographic demonstration of the ovary is successful in most of the active hens with a bodyweight of more than 70g. the origin of the tumours cannot be determined. in connection with this. the well defined structures appeared as large rounded masses of mixed echogenicity. A number of rudimentary follicles within the ovary tissue results in an inhomogenous echotexture. Ovarian neoplasias are distinctly demonstrable by means of sonography. The sonographic complexity of the tumorous tissue allows often a clear demarcation from surrounding structures. In no case were remnants of unchanged testicular tissue demonstrable. Ultrasound-guided biopsy for histopathology is possible but difficult (risk of internal bleeding). Sometimes. Changes in the shape of the follicle were visible due to the movement of adjacent intestinal loops. it is not possible to infer the character of the neoplasias. representing necrotic or cystic areas intensified the inhomogenous impression. The closer the ovaries were located to the body wall. the better the image of the organ was. Histopathologically most neoplasias of the testes are found to be sertoli cell tumours. it is not possible to associate the masses definitely with the testes. occurred as an area of higher echogenicity.echotexture. was not possible. A clear differentiation of inactive ovaries. anechoic or hypoechoic inner structure. which gave it a characteristical appearance of a signetring. in hens it is possible to differentiate follicles or eggs in the oviduct. 441 . In some cases multiple hypo. Histopathologically the masses are often found to be adenocarcinomas or granulosa cell tumours. Evidentially. on the increase in size. The picture of active ovaries is characterised by the presence of follicles in different sizes representing various stages of development. Because of their massive extension. Developing follicles are first seen as round areas with indistinct. A decentrally positioned round area of minor echogenicity was often visible within the yolk. showing the phenomenon of distal acoustic enhancement. is easily recognizable. The ovarial parenchyma. the ova exhibit a distinct separation in echogenic yolk surrounded by poorly echoic perimeter of albumen. Accompanied by massive enlargement of the affected organ. But due to the extension. In advanced stages of development the follicles exhibit the more echoic content of yolk. multiple poorly echoic foci mark necrotic areas within these irregular masses. only visible to some extent between the follicles. Ovarial cysts may occur as single large cysts or as polycystic changes of the ovary. however.to anechoic foci within the masses. Ovarial cysts appear sonographically as clearly defined rounded anechoic compartments. added in the uterus. The hyperechoic shell. in the magnum. More distally in the oviduct. seen as marked focal or diffuse inhomogenous echotexture. Oviduct Irrespective of its functional state. thin-shelled or non-calcified eggs. Depending on the kind of effusion. These are seen as oval to round structures of varying echogenicity. essentially because of the similar echogenicity. HOFBAUER H. In pronounced suppurative processes the echogenicity of the content increases. PITRA C. Moreover. GÖRITZ F. 538 – 555. Pathologic changes of the oviduct are commonly salpingitis. In: RITCHIE BW. KRAUTWALD-JUNGHANNS ME.326. 40: 58 – 64. HOFBAUER H. 3 CITATION INDEX 1. 1996. The sonographic image of advanced salpingitis is. they are sometimes surrounded by a margin of anechoic to hypoechoic fluid. Furthermore. Egg binding: Abnormal eggs are detected most frequently in suspected cases of egg binding. Proc Euro Assoc Avian Vet. et al. Lake Worth: Wingers Publishing 1994. In birds with high grade salpingitis it may be sonographically impossible to make a distinction between the laminated egg and the oviductal wall. In: RITCHIE BW. cysts of the rudimentary right oviduct may be detected. Nephrology. the inflammatory exudates may appear sonographically anechoic to hypoechoic. They show the same sonographic picture as ovarian cysts. HARRISON G and HARRISON L (eds): Avian Medicine – Principles and Application. Furthermore. Lake Worth. recognizable by increased thickness of the oviduct wall. roughness of the egg shells cannot be demonstrated in most cases.41. 3. 4. Lowgrade oviductal inflammation processes are sonographically indistinguishable. which gives the laminated egg a sonographic appearance of onion layers. 5. LUMEIJ JT. particularly in the sectional view. HARRISON G and HARRISON L. MCMILLAN MC. Beitrag zur transkutanen Ultraschalluntersuchung des aviären Urogenitaltraktes. Vet Rad Ultrasound 1999. Egg binding in association with fully formed. Transcutaneous ultrasonography of the avian urogenital tract. Transintestinal ultrasonographic sexing. Florida: Wingers Publishing 1994. malformed eggs as well as eggs with destroyed shells are sonographically assessable. The inflammation of the oviduct is frequently accompanied by an increased incidence of laminated eggs. Giessen: Doctoral thesis. 246 . HILDEBRANDT T. Imaging Techniques. 2. Jerusalem 1995: 37 . In some cases salpingitis may be found after egg binding surgery and following hormonal therapy. correctly shelled eggs does occur quite often in birds. Laminated eggs are clearly demonstrated sonographically. Because of their high echogenicity. 442 . (eds): Avian Medicine – Principles and Application. the unchanged oviduct is often sonographically indistinguishable from the surrounding abdominal structures (intestines). due to different densities of the deposited material. Proc Assoc Avian Vet. KRAUTWALD-JUNGHANNS. An den Tierkliniken 17. Portland 2000: 421 . E. 8. Atlas of Radiographic Anatomy and Diagnosis in Cage Birds. KOSTKA VM. Hamburg and Berlin: Paul Parey 1992. Avian Vet. HUMMEL G. In: RITCHIE BW.6. Orlando 2001: 349-357. Diagnosis and management of female avian reproductive diseases. KRAUTWALD ME. Theriogenology. et al.uni-leipzig. 1992. 04103 Leipzig. University of Leipzig. 9. JOYNER KL. 10. TELLHELM B. Lake Worth: Wingers Publishing 1994: 748-804. Dipl ECAMS Clinic for Birds and Reptiles. Proc Assoc. Radiology of the avian patient – Y2K and beyond. (eds): Avian Medicine – Principles and Application. Gießen: Doctoral thesis. Prof DrMedVet.de 443 . Germany Email: krautwald@vmf. Röntgendiagnostik des aviären Skelettsystems. AUTHORS ADDRESS M. BOWLES HL. HARRISON G and HARRISON L. NEWELL SM. 7.424. . European Association of Avian Veterinarians Conference POSTERS . . 6% in the formulated-diet group. KEYWORDS Diet – Bodyweight– Intestinal flora – Budgerigar ABSTRACT Two groups (n=22) of budgerigars (Melopsittacus undulatus) were maintained under identical conditions with the exception of the diet for 6 months. HARRISON 2003). J. Vetsuisse Faculty. 447 . Keller. med. Average bodyweight of the seed-diet budgerigars remained constant during the trial. Faecal examinations represent an important diagnostic tool in the clinical evaluation of diseased birds. On day 0. P. Following an increase of the formulated diet bodyweights increased. vet. but showed that the composition of the food might have an influence on the intestinal flora (GLUNDER 2002). ornithogaster 1 INTRODUCTION Clinical signs in birds are unspecific and diagnosis has to rely heavily on additional clinical investigative methods. The other group received a commercial formulated diet for budgerigars plus carrots. especially E.5g (16%) on day 15 compared to day 0. The evaluation of the faecal samples revealed no significant differences between the both groups with the exception of Macrorhabdus ornithogaster which was found in 7. Dr. One group was offered a commercial seed mixture plus carrots and a mineral supplement. Hatt. University of Zurich.Division of Zoo Animals and Exotic Pets. coli is not part of the normal intestinal flora of granivorous birds.7% of samples in the seed-diet group versus 36. Dr. The bodyweight of the formulated-diet group dropped after diet-changing to a minimum average of 41. Fischer. 20. In healthy granivorous psittacine birds gram negative bacteria. Switzerland INFLUENCE OF A SEED DIET VERSUS A FORMULATED DIET ON BODYWEIGHT AND INTESTINAL FLORA IN BUDGERIGARS (MELOPSITTACUS UNDULATUS) OVER A SIX MONTH PERIOD I. do not belong to the normal intestinal flora (FIENNES 1959.-M. vet. med. 35. med.. 15. Dr. vet.. The results of this study suggest that transition of diet in budgerigars has to be carefully monitored and that the different diets showed no influence on the intestinal flora with the exception of M. Prof. A recent study supported the assumption that E. coli. 90 and 180 days each animal was weighed and faeces were analysed microscopically with Gram stain. Switzerland). Basel. The phosphorus concentration was measured with an autoanalyzer (Cobas Mira. The evaluation of the faecal samples revealed no significant differences between both groups. At day 90 the average bodyweight (44. with the exception of Macrorhabdus ornithogaster which was found in 7.9g) dropped again (9. 90 and 180 days each animal was weighed and on day 90 and 180 faecal material was smeared in a thin layer onto a slide. The calcium concentration was measured by atomic absorption spectrometry (SpectrAA-20. The required amount of the formulated diet was offered with continuous decrease of the seed fraction. Roche. France) using the procedure described by the manufacturer. One bird died on day 16 due to emaciation as confirmed by necropsy. Jordi AG. BioMérieux. The other group was changed to a commercial formulated diet (Harrison’s Bird Diet Adult Lifetime Fine Grind.5 g FM/bird/d). 15. 3 RESULTS The average bodyweight of the pellet-diet group dropped after diet-changing to a minimum average of 41. Average bodyweight of the seed-diet budgerigars remained constant during the trial. Zug. Switzerland) with a colorimetric method. Gram-negative rods were found in 25. Varian. Statistical analyses were performed with ANOVA.5 g FM/bird/d) and a mineral supplement (Quikon. In three to five uniform optical fields the percentage of Gram positive rods.1g.7g) without change to the diet. Germany) (4g FM/bird/d. Changing to the formulated diet took place over a period of 14 days. One group was offered a commercial seed mixture ad libitum (90 Hammer.6% in the formulated diet group. Additionally. Firma Quiko. Bocholt. Each sample was evaluated by different persons. several fields were scanned for the presence of yeasts and their percentage of budding. 448 . The slides were analysed under 1000x magnification in oil immersion by scanning several optical fields to evaluate the uniformity.7%). Switzerland) plus carrots (0. Gram positive cocci and Gram negative rods were recorded and the average was determined. 20.1). The formulated diet and the seed diet were analysed for nutrient contents (Tab. No yeasts were diagnosed.6% of the samples. SWV English budgerigar-food.7% of samples of seed diet group versus 36. crude fat and crude fibre were determined using standard procedures for Weender analysis. The contents for crude ash.e.5g (i.9%) compared to day 0 and subsequently increased again by day 180 (49. a decrease of 16%) on day 15 compared to day 0. The faecal smears were stained with Gram’s solution (Color Gram 2. manufacturer recommendation) plus carrots (0. 35.2 MATERIAL AND METHODS Two groups including 22 randomly selected 2-year-old budgerigars (Melopsittacus undulatus) each were housed under identical conditions for 6 months with the diet being the only difference.Signau. Germany). crude protein.1%) over Gram-positive cocci (14. Following an increase of the formulated diet of 50% bodyweights increased by day 35 to 48. Avifood. Gräfelfing. and for Macrorhabdus ornithogaster. On day 0. Grampositive rods dominated (79. 109: 266 .5 75 17. A possible explanation for the decrease in bodyweight at day 90 in the formulated diet group could not be found. Furthermore it is not known if the vitamin and mineral-supplement had an effect. HARRISON G. Exotic DVM. Proc Zool Soc London 1959. 4.8 4 DISCUSSION The present study shows that transition of seed fed budgerigars to a formulated diet is possible. 4.13. Values are expressed on a fresh matter (FM) or dry matter (DM) basis. 2003. 132: 129 . Dtsch Tieraerztl Wochensch 2002. 2003. Possible reasons may be that the durations of both studies were not comparable or that the different number of animals had an influence on the interpretation.0 0.0 96 19. GLUNDER G. 4.11.1 4.6: 10 . Influence of diet on the occurence of some bacteria in the Intestinal flora of wild and pet birds.5 171 21.6 Seed diet (dehulled) 100.270.4 36. Additionally it may be that the diets and species used in the study of Stanford (2003) and this study may be too different to allow comparison. Preliminary Field Study of Fecal Gram’s Stain Results in Two Free-Ranging Australian Parrot Species. STANFORD M.3 0. The observation that the formulated diet did not have a significant effect on intestinal flora differs from another study in 100 African grey parrots (Psittacus erithacus) (STANFORD 2003). Report of the Society’s Pathologist for the Year 1957. 5 CITATION INDEX 1. FIENNES T. but that bodyweights should be carefully monitored during transition and amounts fed to birds may have to be higher during transition than for subsequent maintenance. 449 . ornithogaster may be the stress of transition to the formulated diet. Exotic DVM.4 5. 3.Table 1. 2.3 4.6 171 29.0 0.6 32. Analysis of the diets fed to budgerigars (Melopsittacus undulatus).146. Unit Water Crude ash Crude protein Crude fibre Crude fat Metabolizable Energy Calcium Phosphorus Sodium g/kg FM g/kg Dm g/kg Dm g/kg Dm g/kg Dm MJ/kg Dm g/kg Dm g/kg Dm g/kg Dm Pelleted diet 77.6: 12 . Effects of Dietary Change on Fecal Gram’s Stains in the African Grey Parrot. A possible reason for the increased occurrence of M. med. vet. Switzerland. University of Zurich. Email: ifischer@vetclinics. 8057 Zurich.ch 450 . Division of Zoo Animals and Exotic Pets. Dr. Winterthurerstrasse 260. Vetsuisse Faculty.unizh.AUTHORS ADDRESS Isabelle Fischer. Ticks . We also found that the survival rate after 2-3 years of unbanded king penguin chicks with tags is about twice as large as that reported in the literature for banded chicks at different colonies or years. DVM.org 451 . Le Bohec. Gauthier-Clerc. but the significance of the data relies on the assumption that there is no bias introduced in this type of long-term monitoring. PhD. After five years using an automated system of identification of king penguins implanted with electronic tags. Gauthier-Clerc. France Email: gauthier-clerc@tourduvalat. DVM.P.Reproduction ABSTRACT Changes in seabird populations. lower breeding probability and lower chick production. Le Maho KEYWORDS Penguins .C. AUTHORS ADDRESS M. Gendner. France EFFECTS OF FLIPPER BANDS ON FREE-LIVING PENGUINS M. J-P. F – 13200 Arles.Borrelia burgdorfer .E. This often implies large-scale banding to identify individual birds. PhD Station Biologique de la Tour du Valat. offer a unique opportunity for investigating the impact of fisheries and climatic variations on marine resources.E-CNRS Strasbourg. C. Le Sambuc. we can report that banding results in later arrival at the colony for courtship in some years. Y. and particularly of penguins. org. Borrelia burgdorferi antibodies were detected in 14% of tick-infested adults and in 6% of chicks sera. i. one group breeding in an infested area and the other in a non-infested area. the haematocrit level and the behaviour during incubation according to tick infestation. We compared the body mass. France Email: gauthier-clerc@tourduvalat. PhD KEYWORDS Penguins .Station Biologique de la Tour du Valat Arles. The level of infestation varied between years. We compared two groups of penguins carrying individual subcutaneous electronic tags. DVM. DVM. We have studied the effects of tick parasitism by Ixodes uriae in a colony of king penguins (Aptenodytes patagonicus) at Possession Island during three breeding seasons. Contrary to what might have been expected.e. France INFESTATION BY TICKS IXODES URIAE IN PENGUIN COLONIES M. during the incubating period. The survival was not different over the 32 following months between both groups.Reproduction ABSTRACT High densities of penguins in their colonies and the continuous use of these sites over consecutive reproductive periods increase the risk of development of tick populations.Borrelia burgdorferi . Le Sambuc. F – 13200 Arles. Penguins showed a lower incubating success in infested areas during a year of high infestation. PhD Station Biologique de la Tour du Valat.Ticks . Tick feeding activity coincided with the periods when adult penguins stayed ashore for six days or more. Gauthier-Clerc. In an infested area. This duration corresponds to the duration of a tick meal on the host. Gauthier-Clerc. We investigated the prevalence and periods of tick infestation during the one-year breeding cycle of penguins. the haematocrit or the behaviour of incubating penguins. Tick infestation had no effect on the body mass. 452 . individuals seen with ticks had a lower breeding success in rearing a oneyear old chick than those seen without ticks. AUTHORS ADDRESS M. The effects of tick parasitism on penguin breeding performance were assessed from photographs of the colony and with an automatic penguin identification system. birds breeding in the infested area did not show a lower fidelity to that area the next breeding season in comparison to birds breeding in the non-infested area. the provided qualitative result is often inconclusive: what is the meaning of the presence of the detected agent? For example. 1 INTRODUCTION Although clinical signs in affected birds can lead to a presumptive diagnosis. France REAL-TIME PCR FOR DIAGNOSIS OF THREE COMMON INFECTIOUS DISEASES IN CAGED BIRDS : CHLAMYDIOPHILOSIS. Diagnosis testing can be used to confirm or infirm a clinical suspicion and it makes possible to the veterinarian to prescribe a suitable treatment to the bird. 453 . through some clinical examples. BEAK AND FEATHER DISEASE AND AVIAN POLYOMAVIROSIS S. MS KEYWORDS Real-time PCR . is it an acute or a subclinical infection? Real-Time PCR is an advanced PCR-based technology which enables to assess the quantity of one specific agent in various biological samples. provides helpful results either for the approach of a clinical case or for the detection of asymptomatic carriers. definitive diagnosis often involves demonstration of virus presence. A quantitative tool can make interpretation easier. Direct diagnosis with a quantitative approach produces a conclusive result when others assays may sometimes not provide any solution for avian practitioners. asymptomatic carriers have to be identified to prevent dissemination and economic losses in facilities and to reduce the zoonotic risk with Chlamydophila. as these diseases are very contagious.Toulouse.Infectious diseases . Whereas these assays can be sensitive and specific. Nucleic acid detection of a pathogen.Caged birds . we describe this technique and. Furthermore. The Polymerase Chain Reaction (PCR) is an effective method for the detection of these three different pathogens. the interest to perform Real-Time PCR analyzes in birds veterinary medicine is shown. as PCR. Here. Lafon.Scanelis.Pathogen quantification ABSTRACT Chlamydophila psittaci. beak and feather disease virus (BFDV) and avian polyomavirus (APV) are common pathogens in psittacine birds. Some real-time PCR assays for caged-birds infections and their interest are described bellow. a specific probe (100 nM) and primers (600 nM) for 60 cycles. The extraction yield and the absence of PCR inhibitors were checked with a specific real-time PCR. EDTA blood samples and feather calamus have been screened for BFDV and APV and cloacal swabs for C. Amplification curves show that very different initial pathogen loads lead to equivalent amounts of PCR products after 60 PCR cycles. 3 RESULTS 3.5 % agarose gel. 454 . 3. the detection of asymptomatic but infected birds is improved. BFDV conventional PCR: a specific assay was used to amplify a fragment of BFDV genome. 2. the relative quantification of the viral load at two different times provides very interesting information about disease evolution and/or the efficiency of any treatment. The electrophoresis result confirms this finding: end-point quantification is not accurate. PCR products run in 1. The limit of detection of PCR was assessed on quantified templates. 3. an improved PCR-based technology. Realtime PCR: the TaqMan® system. 1 A higher sensitivity for a best detection of APV healthy carriers As only 10 copies of APV genome lead to a positive result with our assay. PCR Internal Control: a synthetic DNA was added in each sample at the first step of the DNA extraction. the lower the Ct value is. 2. The more the initial amount is high. Also any trace of virus can be detected. real-time PCR does! Real-time PCR in TaqMan® system is a PCR with internal probes for the quantification of PCR products. 2. 1 Whereas conventional PCR can not provide quantitative data. The Ct (Threshold cycle) value is the number of cycles necessary to reach a defined fluorescence threshold. 3 Viral load monitoring for BFDV: transient viraemia or disease progress? For one bird. allowing prevention of virus dissemination in the breeding. 2 A tool to interpret chlamydophilosis testing results The bacterial load can be helpful to assess the role of the detected Chlamydophila in the clinical signs observed.2 MATERIAL AND METHODS DNA extraction: DNA was extracted from each field sample with High Pure PCR Template Purification Kit (Roche). psittaci. was used on an ABI7900 SDS (Applied Biosystems). 2 Interest of sensitivity and pathogen load in veterinary medicine: examples 3. Conditions: 2X UDG Platinum® Quantitative PCR (Invitrogen). 3. Specificity was checked on various pathogens. Research and Development Scanelis . France Email: soph. Compared to conventional PCR.4 DISCUSSION There are many applications of the real-time PCR in the veterinary field.com 455 . both improving the quality of the result. 31076 Toulouse Cedex 3. MS.23.National Veterinary School of Toulouse . AUTHORS ADRESS S. this technology increases sensitivity and allows quantification. Lafon. chemin des Capelles.lafon@scanelis. The increase of specificity and the end of carry-over problems are two other major sides which bring false-positive results with conventional PCR to an end. Larhantec KEYWORDS Oil spill . However. not only hydrocarbons but also metals such as nickel. PhD. We suggest that vanadium may be not a good marker of acute oiling but may be better to mark a chronic toxicity. Ecole Nationale Vétérinaire Nantes. we present a study about the vanadium content of organs of oiled birds since the Erika oil spill in France. Vanadium was measured by atomic absorption spectrophotometry in liver and kidney of several oiled seabird species. Pouliquen DMV. The toxicity of petroleum is related to its composition. A certain amount of data is beginning to be accumulated concerning its presence in marine organisms (MIRAMAND and FOWLER 1998). probably in relation to their diet. Moreover. Kammerer. ECPVT Diplomate.Ecotoxicology . The aim is to study the differences between species and between the types of oil (with different vanadium value). It also has a specific toxicity (cellular and DNA toxicity). The concentrations were not higher in oiled birds than in dead birds found on the beaches without visible traces of petroleum. there appears to be differences between origins of birds (Atlantic vrs Channel) but not between types of oil. DMV. stranded on French Atlantic and Channel coasts that had died in wildlife care Centres. aluminium or vanadium. France VANADIUM CONTENT OF OILED BIRDS : IS IT A GOOD MARKER FOR OIL EXPOSURE ? S. Le Dréan-Quenec’hdu. PhD. M. DMV. H. 456 .Wildlife Centre. but its metabolism in other animal species is not well known. 1 INTRODUCTION Seabirds that have been victims of accidental release or operational discharges from ships suffer primarily from the physical action of petroleum. M. In this poster. PhD. The aims of this study are to compare the vanadium content of oiled birds form different species (with different diet) and form different origin (different hydrocarbons). during the winters of 19992003. we showed that there were differences between species.Seabirds ABSTRACT Vanadium is a trace element. contaminating oil products. p<0. in parallel with some hydrocarbons of feathers. 3 RESULTS The vanadium content of livers according to species varied between 26ng/g and 131ng/g. The procedure has been described with details in KAMMERER et al. and thus the possible physiological and toxicological consequences seem however limited enough for oiled seabirds.247. USA).2 MATERIAL AND METHODS The analysed birds died naturally in wildlife centres during the Erika oil spill. 457 . Moreover. It would be interesting to complete it by other analyses for various types of hydrocarbon with content vanadium known and by dosages. during summer 2001. it is possible that this variability comes from the variability of exposure of birds. 4 DISCUSSION The values for concentrations of vanadium in the liver of birds found in the literature vary from 18 to 38 ng. Moreover. the variability of data was also higher. The hypothesis we made was that the birds wintering in Channels wintered in “polluted zones “. 2001-2002 (chronic pollution). The data showed an important individual variability. 20002001. However. the Prestige and Tricolor oil spills. p>0. The vanadium contents were significantly higher for birds that died during winter 2002-2003 than during previous winter. were taken and separately stored at –20°C in polyethylene flasks. liver and kidney were also collected on seven other Allcidae found dead without visible traces of petroleum.1374.006). Vanadium concentrations were determined by using an atomic absorption spectrophotometer 1 100 B (Perkin-Elmer. the birds coming from Channel showed higher vanadium content than birds coming from Atlantic sea. The vanadium resumption.g-1 for waterbirds living in polluted zones. Norwalk.05.092.1 in 3 ppm (PULS 1994) and from 100 to 900 ng. There were also significant differences between types of hydrocarbons (F=4. CT. Our results are widely superior to the values for not exposed poultry but the lack of reference value makes difficult the interpretation. in the skeleton. without gall-bladder. The relation between vanadium content of oil sampled on feather and of livers was not significant but the sample is low (r2=0. These birds were used as “controls”. Our study does not allow the assertion that the vanadium represents a good biomarker of the pollution by oil to the birds. Furthermore control birds showed vanadium contents higher than oiled birds (except «Tricolor» birds).001).g-1 (fresh weight) for poultry receiving a food with a vanadium content of 0. p=0. n= 8). in fishing-nets in the Channel. Whole kidney and liver. Indeed the bone could be a better marker of the exposure. However. even control birds that were not oiled. within the same sort. There were significant differences between species (t guillemot vrs kittiwake = -3. (2004). Clearbook 1994.fr 458 . LE DRÉAN-QUENEC’HDU S. Bioaccumulation and transfer of vanadium in marine organisms. 2. Sherpa International. PULS R. DVM. Ecole Nationale Vétérinaire. 333: 295 . Liver and kidney concentrations of vanadium in oiled seabirds. BP 40706. Atlanple-Chantrerie. In: NRIAGU (ed): Vanadium in the environment. POULIQUEN H and LARHANTEC M. The Sci Total Environ 2004. AUTHORS ADRESS Le Dréan-Quenec’hdu S. MIRAMAND P and FOWLER SW. PhD Wildlife Centre. Mineral levels in animal health 2nd ed. Part 1: Chemistry and biochemistry. 1998. 3. 44307 Nantes cedex 03. Canada. France Email: sldq@club-internet. New York: John Wiley and Sons Inc. 167 . MASTAIN O.5 CITATION INDEX 1.301.197. mallards).Toxicology . The release rate will be also shown. 2 CAUSES OF CARE Hundred and twenty sorts were so taken in among which 106 sorts of birds.Infectious disease .Seabirds . KEYWORDS Wild birds . In Nantes. hunting. The causes of the collect will be analyzed according to season and to species. The sea birds as the guillemots are welcomed because of oil. the wildlife centre of the National Veterinarian School of Nantes welcomes these injured animals. The waterbirds are 459 . 1 INTRODUCTION The increasing human activity leads numerous modifications of the environment and assaults the balance of the ecosystems. birds of prey (common buzzard. France CAUSE OF MORTALITY OF WILD BIRDS COLLECTED IN A WILDLIFE CENTER IN FRANCE : EXAMPLE OF NANTES WILDLIFE CENTER Le Dréan-Quenec’hdu S. All these animals are examined and looked after with for objective their insertion or reintegration in their environment. DVM. 44307 Nantes cedex 03. PhD. Since 1985. Lambert O.Trauma ABSTRACT More than 1000 wild birds are collected each year in the wildlife center of the National Veterinary School of Nantes (France). Most of them are raptor and sea birds. The causes of entrance vary according to the sorts. in the respect for charter of the French Union of Wildlife Centres (UFCS). Risi E. electrocution. Every day. gulls. the animals are mainly birds (94% in 2003). which alone could not survive. numerous animals of the wild fauna are victims of poisoning. plate glass windows) and of diseyrie in breeding period.BP 40706. The most frequent sorts are the sea birds (guillemots. Ecole Nationale Vétérinaire Atlanple-Chantrerie. barn owl). common kestrel.Wildlife Centre. The collisions with vehicles or electric lines are the most frequent causes of entrance for birds of prey (58 barn-owls entry for collision with a car in 2003).Raptors . DVM. accidents (road. The release rate is very variable according to the species and according to the causes of entrance. Atlanple-Chantrerie. in spring. to dive). 46% died in care and 41% were released. It is hospitalised in rooms adapted to its species. For example. Finally. PhD Wildlife Centre. Ecole Nationale Vétérinaire. AUTHORS ADRESS Le Dréan-Quenec’hdu S. several decisions can be taken. it is also euthanised: This obligation to look after animals savage in the objective to release them in the wild is registered in the rule and in the charter of the French Union of the Wildlife Centres. - In 2003. BP 40706. 13% of the animals were euthanised. only 12% of the barn owls were released. it will be transferred in another centre. If the animal cannot be taken care on the centre (for example seal). more than 85% of the little owls were released.essentially welcomed because of illness (botulism). it is taken care: he can then benefit from medical care and / or surgical. the animal cannot be released in its natural environment (example of a bird of prey which we should amputate of a wing or which would have become blind). in 2003.fr 460 . The causes of entrance also vary according to the season: oiling is more frequent in winter (because reach the wintering populations). 3 RELEASE RATE According to the state of the animal. If the veterinarian thinks that the animal has chances to be able to be released in the wild after the care. adapted to its species. the centre welcomes mainly young birds fallen from the nest. Then it benefits from a phase of rehabilitation / rehabilitation (for example the sea birds are put in swimming pool to re-learn to live on the water. If we think that in spite of the care. If the animal presents too grave wounds after which we cannot look. it is released after having been ringed. it is euthanised. DVM. On the other hand. 44307 Nantes cedex 03 France Email: sldq@club-internet. A. KRAUTWALD-JUNGHANNS et al. 1993. 2 MATERIALS AND METHODS Two adult male and female pigeons were used in this study. but the use of computed tomography (CT) technology is increasing. De Rycke2. The present study describes the CT anatomy of the head. General anaesthesia was 461 . No references of tomographic anatomy for pigeons are currently available. H. compared to anatomical sections of frozen pigeons. Ghent University. Department of Medical Imaging2. The present study describes the CT anatomy of the head. as this technique avoids superposition of overlying structures and provides a better soft tissue differentiation than classical radiology. 1998a. F. 1 INTRODUCTION Routine radiography is still the most frequently performed imaging modality. To avoid movement and stress during CT examination. Bacteriology and Avian Diseases1. Several reports have already described the use of CT in psittacines. van Bree2 KEYWORDS Pigeon – Anatomy – Computed tomography ABSTRACT Routine radiography is still the most frequently performed imaging modality in avian patients. Faculty of Veterinary Medicine. Gielen2. compared with anatomical sections of frozen pigeons. as this technique avoids superposition of overlying structures.Department of Pathology. the birds were anaesthetized. I. thorax and abdomen of the pigeon (Columba livia). Belgium COMPUTED TOMOGRAPHIC ANATOMY OF THE PIGEON (Columba livia) A. Van Caelenberg2. 1998b. raptors and anseriformes (BARTELS et al. Martel1. Merelbeke. The birds appeared healthy at clinical examination on the day of the study. OROSZ and TOAL 1992). 2000. L. but the use of computed tomography (CT) technology is increasing. Pasmans1. thorax and abdomen of the pigeon. GUMPENBERGER and HENNIGER 2001. and HENNINGER W. However. 2. Comparative studies on the diagnostic value of conventional radiography and computed tomography in evaluating the heads of psittacine and raptorial birds.108. Evaluation of the lower respiratory tract in psittacines using radiology and computed tomography. Principles and concepts of computed tomography. CT examinations were done using a CT Pace scanner (General Electrics) with the birds positioned in ventral recumbency. KRAUTWALD-JUNGHANNS ME. 23: 399 . but this is not a significant problem.180. 5. The image acquisition time was 15 minutes. 462 . were obtained without overlap. GUMPENBERGER M. BARTELS T. J Avian Med Surg 1998a. For example to distinguish the intestinal tract from other viscera. CT images provided good anatomical detail of the internal structures. Vet Radiol Ultrasound 1993. 3. 34: 382 .390.157. KRAUTWALD-JUNHANNS ME. SCHUHMACHER F and TELLEHLM B. PORTMANN S. The use of conventional radiography and computer-assisted tomography as instruments for demonstration of gross pathological lesions in the cranium and cerebrum in the crested breed of the domestic duck (Anas platyrhynchos forma domestica). such as the gonads and the cranial pole of the kidneys. Veterinary Clinics of North America: Small Animal Practice 1993. Despite these small limitations. Sem Avian Exotic Pet Med 2001.414. 4 CITATION INDEX 1. this differentiation is visible on the sagittal scans. KOSTKA VM and DÖRSCH B. 3 RESULTS AND DISCUSSION The results of the study will be shown in sequential matched photographs of the anatomical sections with their corresponding CT images (see poster). the euthanised pigeons were frozen at -20°C and sectioned into approximately 10 mm thick slab sections using an electric planner. Avian Pathol 2000. Transverse and sagittal sections were made. Transversal CT images may be unable to differentiate between soft tissue organs in close proximity. Effects of breathing motion are seen on occasional transverse images. et al. 10: 174 . The use of computed tomography in avian and reptile medicine. 29: 101 .induced and maintained with isoflurane and oxygen by face mask. Afterwards. HATCHCOCK JT and STICKLE RL. 4. CT is a useful tool in the study of avian anatomy. KRAUTWALD-JUNGHANNS ME. A 3-second scan time was used. Twenty-six contiguous 2 mm thick transverse CT-slices from the head and heart region. 12: 149 . and 33 contiguous 5 mm thick transverse scans beginning from the thoracic inlet and proceeding to the cloaca. CT images were made using 100 mA and 120 kV. barium sulphate can be used. Contrast enhancement of the soft tissues during total body CT would help to identify and differentiate various organs. Anatomical sections were photographed and compared to the corresponding CT images. Eighteen 5 mm thick sagittal slices were taken. 7. 9820 Merelbeke. Salisburylaan 133.Arten mit hilfe der rekonstruktiven röntgentransmissionstomogr aphie.70. 26: 61 . SCHUHMACHER F and SOHN HG.Martel@ugent. OROSZ SE and TOAL RL.6. Untersuchungen am unteren respirationstrakt von Psittacinae. Martel.46. J Zoo Wildlife Med 1992. Tomographic anatomy of the golden eagle (Aquila chrysaetos). KRAUTWALD-JUNGHANNS ME. AUTHORS ADDRESS A. Faculty of Veterinary Medicine.und Amasonae. PhD Clinic for Avian and Exotic Animal Diseases. Tierärztl Prax 1998b. 23: 39 . Belgium Email: An.be 463 . DVM. Exposure – Anticoagulant rodenticides – France ABSTRACT An epidemiological survey was performed in Loire Atlantique (France) on raptors and water birds which did not present with clinical signs or lesions of poisoning. Pouliquen. or secondary by eating contaminated preys (SHORE et al. carrots. to five anticoagulant rodenticides (brodifacoum. coumafen. reflecting its wide use in Loire Atlantique against coypus. 1 INTRODUCTION Many anticoagulant rodenticides are used in field-treatments in France. The aim of this study was to evaluate possible exposure of these non-target wildlife species to anticoagulant rodenticides used against rodents. L’Hostis KEYWORDS Raptors – Water birds . bromadiolone was the most frequent detected or quantified in the liver of birds. Lambert. either directly by eating contaminated baits. 11 common buzzards (Buteo 464 . O. The exposure of raptors was large and significantly greater than the exposure of water birds. Among the 5 anticoagulants tested. 2 MATERIALS AND METHODS Thirty raptors and 28 water birds from Loire Atlantique (France) collected in the wildlife centre of the national veterinary school of Nantes were included in this study : 15 diurnal raptors [4 kestrel falcons (Falco tinnunculus). Larhantec. The aim of this study was to evaluate the exposure of on target species. they might have a negative impact on non-target species. C. M. France DETERMINATION OF THE POSSIBLE EXPOSURE OF RAPTORS AND WATER BIRDS TO FIVE ANTICOAGULANT RODENTICIDES IN LOIRE ATLANTIQUE (FRANCE 44) H.Wildlife Center National Veterinary School of Nantes. principally due to their feeding habits. apples or cereals. such as raptors and water birds. bromadiolone. M. 1996. 1997). Thorin. BERNY et al. coumatetralyl and difenacoum). Despite the fact that they are used against rodents. FREESTONE P and KITCHENER AC. 91: 279 . 187 . BURONFOSSE T. exposure of raptors is significantly greater than exposure of water birds. 15 nocturnal raptors [10 barn-owls (Tyto alba). The anticoagulant most frequently detected or quantified in liver was bromadiolone.1828. Bull Environ Contam Toxicol 2003. 70: 34-40. (1999. Identification and quantification of brodifacoum. 5. J Wild Dis 1999. coumafene in 5 birds. BURONFOSSE F. 4 DISCUSSION Our results show that the raptors exposure to anticoagulants rodenticides is large and similar findings were reported by STONE et al. 4. Furthermore. probably because of their feeding habits (MERSON et al 1984). OKONIEWSKI JC and STEDELIN JR. Poisoning of wildlife with anticoagulant rodenticides in New York. due to its regular and common used in wetlands from Loire Atlantique in the form of contaminated carrots or apples baits against coypus (Myocastor coypus).buteo)]. Secondgeneration rodenticides and polecats (Mustela putorius) in Britain. MERSON MH. Anticoagulant rodenticides and raptors: recent findings from New York. STONE WB. difenacoum in 8 birds. 35: 1817 . LAMARQUE F and LORGUE G. coumafen.52 µg/g). Chemosphere 1997.193. 1998-2001. J Wildl Manage 1984. BERNY P. For all birds. OKONIEWSKI JC and STEDELIN JR. BYERS RE and KAUKEINEN DE.282. Envir Pollu 1996. BIRKS JDS. the concentration in liver was low (<1 µg/g) except for one common buzzard (coumafene liver concentration = 2 µg/g) and one black coot (coumafene liver concentration = 23. 48: 212 . Bromadiolone was detected or quantified in 15 birds. SHORE RF. coumatetralyl and difenacoum in liver were completed at the Toxicology Laboratory of the National Veterinary School in Nantes.216. 5 tawny owls (Strix aluco)] and 28 water birds [15 mallards (Anas platyrhynchos). 5 CITATION INDEX 1. The liver of each dead bird was sampled and kept at -18°C until analyses. a 4-year survey. 3 RESULTS Anticoagulant rodenticides were detected or quantified in 22 raptors (73%) and 4 water birds (14%). Residues of the rodenticide brodifacoum in voles and raptors after orchard treatment. 2003). bromadiolone. brodifacoum in 4 birds and coumatetralyl in only one bird. STONE WB. 13 black coots (Fulica atra) and 1 common moorhen (Gallinula chloropus)]. 3. 2. 465 . Field evidence of secondary poisoning of foxes (Vulpes vulpes) and buzzards (Buteo buteo) by bromadiolone. Pouliquen Laboratoire de Diagnostic Toxicologique.La Chantrerie.fr 466 . Ecole Nationale Vétérinaire de Nantes. 44307 Nantes Cedex 03.AUTHORS ADDRESS H.risi@wanadoo. Atlanpole . France Email: emmanuel. DVM. Dipl ECAMS C. degenerative. Remple. penguins and swans are particularly susceptible (HUMPHREYS 1996). A foot cast is designed to relieve pressure (a major factor in the initiation of bumblefoot and an obstacle to healing) on prominent weight-bearing pads and redistribute body weight and pressure onto other plantar pedal surfaces. heavy. SUPPORT CUSHION TO FACILITATE WOUND HEALING IN BUMBLEFOOT J. REIDARSON ET AL 1999). poor perching surfaces and increased body weight are factors in development (HARCOURT-BROWN 1996). Large.Pododermatitis – Foot cast – Cushion – Silicone impression material INTRODUCTION Bumblefoot (pododermatitis) is a chronic. Nurse KEYWORDS Birds – Falcons – Bumblefoot – Metatarsal pad – Digital pads . Bumblefoot has been recognized in falcons for centuries. 467 . D. Colorado. and most information on the disease emanates from that avian group. granulomatous disease of the avian foot. Inc. FLEXIBLE. rubber foot cast for birds.Aspen Wing Bird and Animal Hospital. Kentucky Blacksmith Farrier and Veterinary Supply. IN). Indianapolis. made from a composite silicone dental impression material that has been modified for veterinary use as an equine sole support for acute laminitis (K B Cushion SupportR. The disease usually begins by repeated trauma and excessive pressure to plantar pedal skin resulting in devitalization of the epithelial barrier to infection with microbial entry into underlying tissues (REMPLE 1993). RIDDLE and HOOLIHAN 1993. Foot casting has also proven to be of value in preventing bumblefoot from developing in a healthy foot as a result of prolonged weight-shifting due to injury or disease in the contralateral limb (HARCOURT-BROWN 1996). This report describes the construction and application of a soft. Owing to the pressure relief on lesion areas. flexible. REMPLE 1993. foot casting has shown to assist healing and minimize recurrence of bumblefoot following surgical debridement (REMPLE and REMPLE 1987. Loveland. United States of America USE OF A COMPOSITE SILICONE DENTAL IMPRESSION MATERIAL TO CREATE A FORM-FITTING. and lack of exercise. non-perching birds such as falcons.. Bumblefoot occurs nearly exclusively in birds held in captivity. The padding. and compressed to the contours of the foot and proximal digits. The metatarsus is lightly bandaged with a non-adhesive elastic wrap (VetrapR. which is later removed from the cast. Casts were well tolerated by birds because they were comfortable and they allowed full use of feet while being worn. there silicone composite was placed into preformed moulds of falcon’s feet. A small piece of adherent padding (Dr Scholl’s MolefoamR. non-adhering dressing (TelfaR.MATERIALS AND METHODS Step 1: Upon completion of the surgical procedure. plantar side down. while the cast maker works the composite into every “nook and cranny” along the sides of each toe and the metatarsus. and a rough outline of the foot is traced onto the paper. 3M Animal Care Products). they can cause discomfort. At normal room temperature the composite has a moulding time of about 2 minutes and a setting time of 4 minutes. the exterior of the cast is trimmed with a #11 BP scalpel blade to form a neatly fitting ‘half-shoe’. the incision is overlaid with a thin. These falcons all returned for cast removal weeks later after successful hunting with the foot casts still intact! Previous reports have described various rigid casts (REMPLE and REMPLE 1987. The cast is affixed to the foot with vetrap that encircles the toes and cast. approximately the diameter of a piece of chalk. The foot is held motionless. The tracing extends to the ends of each digit. RESULTS AND DISCUSSION The casting method described above is an updated version of the one previously developed at the Dubai Falcon Hospital (DFH). REMPLE 1993. impede pedal function and/or transfer 468 . RIDDLE and HOOLIHAN 1993). and placed onto the foot tracing. and the padding incision spacer is removed from the foot or cast (whatever it has adhered to) and discarded. the bird is lifted out of the foot cast. The mixed composite is formed into a cylindrical shape. When the composite has set (~4 minutes). acts to create a dead space between incision and form-fitting cast. Step 2: An assistant holds the bird and places the bandaged foot. Colgate-Palmolive). Step 3: Equal parts (red and white) of the silicone composite impression material (KB Cushion SupportR) are quickly mixed in the palm of the hand to a uniform pink colour. Next. in a normal standing position on top of a sheet of paper. With the bird sedated. Schering-Plough) measuring the length of the incision X 5 mm wide is laid over the incision area and pressed onto the outside of the vetrap bandage. and presses the foot and toes into the silicone composite. birds were prematurely released to their owners for hunting purposes with foot casts still on. On several occasions. Trimming at these locations allows for unhampered toe flexion while the cast is being worn. Hundreds of silicone foot casts have been used at DFH over the past decade with excellent results. Step 4: The ends of the cast toes are trimmed off just distal to the middle digital pads of digits 3 and 4 and approximately 2/3rds the distance along digits 1 and 2. Although rigid casts relieve pressure on the metatarsal pad. the assistant again holds the leg in a normal standing position. A novel approach to the treatment of bumblefoot in penguins.167. REMPLE JD. A form-fitting. Dip ECAMS Aspen Wing Bird and Animal Hospital. 5. and it addresses all the concerns of rigid foot casts.314.164. Foot and leg problems. 3. HARCOURT-BROWN NH. J Amer An Hospital Assoc 1987. RIDDLE KE and HOOLIHAN J. In: BEYNON PH. FORBES NA and HARCOURT-BROWN NH (eds): Manual of Raptors. In: BEYNON PH.net 469 . COOPER JE.127. 161 . Pigeons and Waterfowl. Colorado. Pigeons and Waterfowl. J Avian Med Surg 1999. CITATION INDEX 1. mould of the foot that cushions and supports the entire foot while selectively relieving pressure from lesion areas. 13(2): 124 . Foot casting as adjunctive therapy to surgical management of bumblefoot in raptorial species. 7. Minneapolis: University of Minnesota Press 1993. Wing and leg problems. United States of America Email: jdremple@earthlink. FORBES NA and HARCOURT-BROWN NH (eds): Manual of Raptors. AUTHORS ADDRESS J D Remple DVM.4.8. Glouchestershire: BSAVA 1996. 23: 633 . The silicone foot cast described above is the latest advancement in an evolution of foot casting. In: REDIG PT. 9.foot pressure to hard cast areas with new bumblefoot lesions ensuing as a result. Minneapolis: University of Minnesota Press 1993. REMPLE JD and REMPLE CJ. In: REDIG PT. HUMPHREYS PN. The finished ‘halfshoe’ forms a flexible. REMPLE JD and HUNTER DB (eds): Raptor Biomedicine. REIDARSON TH. 313 . 163 . Raptor bumblefoot: a new treatment technique.2. Glouchestershire: BSAVA 1996. and BURCH L.639. 154 160. REMPLE JD and HUNTER DB (eds): Raptor Biomedicine. Loveland.6.10. composite-casting method for avian appendages. COOPER JE. MCBAIN J. 11. It could only move by leaning its wings on the floor.Wildlife Centre National Veterinary School of Nantes. 2 radial carpal bones. in a very poor condition. 2 CASE REPORT The young owl was probably 2 to 4 weeks old. The poster presents the clinical aspect of the bird. L’Hostis M DVM. France CONGENITAL ABNORMALITIES ON A WILD YOUNG LONG-EARED OWL (Asio otus) Risi E.Asio otus – Malformation – Abnormalities .Congenital ABSTRACT A wild young long-eared Owl (Asio otus) was brought to the wildlife centre of the National Veterinary School of Nantes. 2 supernumerary metatarsal bones and 2 supernumerary toes 470 . Several anatomical malformations were observed including 6 digits on each foot. showed ataxia and died during the first night. and each wing bore 2 alular digits. 2 alular digists on the wings and 2 incomplete supernumerary tarsometatarsi on each leg. 2 alular digits on both wings. at the bottom of a tree. A radiograph showed the presence of 2 radius on the left wing associated with a marked bowing of the radius and ulna on the left side and a slight bowing of the same bones on the right side. Physical examination showed skeletal malformations. PhD. Three bones were palpated on the left wing. 2 radius on the left wing. probably just under its nest. The bird had been found in the wild. 1 INTRODUCTION In June 2002. a long-eared owl chick (Asio otus) was presented to the wildlife centre of the National Veterinary School of Nantes (France). The bird was a few weeks old. DVM. 2 radial carpal bones on both wings. it was ataxic and unable to walk. The bird was in a poor condition and very weak. Costiou P. radiographs and anatomical preparations to preserve the bones and muscles. DVM. The bird had 6 toes on each foot. PhD KEYWORDS Long-eared owl . BARREIRO et al. 2002). The bird died on the first night after admission and the body was preserved in alcohol for anatomical research. In these 2 cases. One case of congenital dislocation of radius and carpometacarpus was described in a wild tawny owl chick (BARREIRO et al. 2003). polychlorinated biphenyls (FERNIE et al. 3 radius. . Chemical and physical factors are known to be teratogenic in vertebrates including 6-amino nicotinic acidamide. 2003). Wild birds with anatomical malformations are very rarely observed. At necropsy. Absence of alular digits has been described in a wild tawny owl chick (Strix aluco) (BARREIRO et al. 4 radial carpal bones. 12 toes). 4 CONCLUSIONS This case is of interest for several reasons: . the number of radial bones was normal.The large number of skeletal anomalies described (4 alular digits. 2002. 6 metatarsal bones. Unfortunately. 2003) and less frequently an additional digit distant from the foot (FOX 1989). . no internal malformations were observed.around the hind toe (digit I) of both feet. because they certainly died in the wild before being found. 2 bowed radius could be observed and one of them was clearly enlarged. The case described in this poster seems to be the first description of duplicated radius on a bird. The survival of this long-eared owl had probably been allowed by the parents raising up to the point of leaving the nest. elevated temperature during early embryonic development (CROSTA et al. this is the first documented report in a long-eared owl. A peregrine falcon (COOPER 1984) was found with bowing of one radius associated with carpometacarpal anomalies. One case of incomplete duplicated radius was reported in a black vulture (Coragyps atratus) with an ectopic wing hanging from its cervical region (OSOFSKY et al 1990). Extra alular digits are a less frequent anomaly. Radius anomalies are less frequently reported than those of the digits.The bird was found in the wild and did not come from a breeding centre (rarely found. low level of inbreeding). The antebrachium of this third wing consisted of 2 bones. On the left wing. no toxicological investigations were performed in our case. 2002). 2002). One case has been reported in a goshawk (Accipiter gentilis) and a peregrine falcon (Falco peregrinus) (COOPER 1984) with a shortened carpometacarpus and an extra alula on one side only and one case in a common snipe (Gallinago gallinago) with bilateral supernumerary halluces (CROSTA et al. . Up to 9 digits (in a hawk) have been described in raptors (CROSTA et al. each of which was duplicated beginning at mid shaft (OSOFSKY 1990). insulin (COOPER 1984). Few cases have been documented in raptors and pet exotic birds. 2003). a radius and ulna.To our knowledge. this case is the first documented report of radius duplication. CROSTA et al. retinoids (CROSTA et al. 2002).To our knowledge. 2 bowed ulna. 471 . Polydactylism is the most common malformation described in birds of prey and reports describe generally digit I duplications (COOPER 1984. 3 DISCUSSION Anatomical abnormalities are not rare in farm breeding birds (poultry). The aetiology of these anomalies remains unclear but hazardous material related genetic mutation can not be excluded. 5 CITATION INDEX BARREIRO A et al. Congenital skeletal abnormalities in a tawny owl chick (Strix aluco). Avian Dis 2003; 47: 774 - 776. COOPER JE. Developmental abnormalities in two British falcons (Falco spp). Avian Path 1984; 13: 639 - 645. CROSTA L et al. Unilateral pentadactylism in a yellow-shouldered Amazon (Amazona barbadensis). J Avian Med Surg 2002; 16(1): 26 - 30. FERNIE K et al. Reproductive abnormalities, teratogenicity, and developmental problems in American kestrels (Falco sparverius) exposed to polychlorinates biphenyls. J Toxicol Environ Health A 2003; 66(22): 2089 - 2103. FOX NC. A unilateral extra digit in a wild commom buzzard (Buteo buteo). Avian Path 1989; 18: 193 - 196. OSOFSKY SA et al. An ectopic wing in a wild black vulture (Coragyps atratus). Avian Dis 1990; 34: 765 - 769. AUTHORS ADRESS Emmanuel Risi Centre Vétérinaire de la Faune Sauvage, Ecole Nationale Vétérinaire de Nantes, Atlanpole, La Chantrerie, BP 40706, 44307 Nantes cedex 03, France Email:
[email protected] 472 Wildlife Centre National Veterinary School of Nantes, FRANCE SUCCESSFUL TREATMENT OF A CERVICAL RHABDOMYOSARCOMA ON AN INDIAN FANTAIL PIGEON (COLUMBA LIVIA) BY SURGICAL RESECTION Risi E, NGuyen F, Albaric O, Abadie J KEYWORDS Bird - Pigeon, - Rhabdomyosarcoma - Surgery, - Neoplasm ABSTRACT A domestic Indian fantail pigeon (Columba livia) was presented with a large ulcerated mass on the cervical region. After clinical, endoscopical and radiological examinations, it was decided to treat this mass surgically. The neoplasm was diagnosed as a rhabdomyosarcoma by immunohistochemistry. This poster presents the clinical aspect of the tumour and the successful surgical treatment. The pigeon was still alive two years after surgery. This case is the second published case of rhabdomyosarcoma on a pigeon and the first published case of a successful treatment. 1 INTRODUCTION Rhabdomyosarcoma is a rare malignant neoplasm in birds, previously described in hens, budgerigars, a pigeon and a vulture. All previous cases were observed at postmortem exmination, with metastasis in the lung, spleen, liver and/or kidneys. Here we describe a case of rhabdomyosarcoma in an Indian fantail pigeon (Columba livia), which was successfully treated by surgical resection. 2 MATERIAL AND METHODS A 1-year-old Indian fantail pigeon was presented with a cervical mass of 4 weeks duration. Since 3 days, bleeding and head tilt were observed by the owner. The bird was not able to eat any more, because of the weight of the mass and the abnormal position of the head. The mass was located in the cervical region, near the crop. It was mobile, red, firm, round, 3 cm2, partially plucked and ulcerated. No other abnormality 473 was observed. By endoscopy, the internal wall of the crop was normal and seeds were present inside. Radiographs of the body in a lateral and a dorsal recumbency did not reveal any lesion, in particular any metastasis. A surgical resection of the mass was performed. The bird was anaesthetised with isoflurane by face mask and then maintained after intubation (isoflurane 5%, 02 1,5 L/min, then 1,5%, 02 1 L/min). The area was gently plucked and disinfected with iodine povydone (“Vetedine”). A sterile gauze was used as drape. Dissection was performed around the mass and along the crop wall. The crop wall was very thin and seeds were seen through. Two veins and one artery were ligated before removing the mass. After checking the absence of crop lesion, the skin was sutured with a interrupted suture pattern. Antibiotics were given for 5 days (amoxycillin-clavulanic acid “SYNULOX” 125 mg/kg bid PO). The biopsy was fixed in formalin 10%, embedded in paraffin, cut and stained with haematoxylineosin (HE). Serial sections were used for immunohistochemistry using the biotinavidin-peroxidase technique, with anti-vimentin (1:1000, Dako M0725), anti-desmin (1:200, Dako M0724), anti-smooth muscle actin (1:1000, Dako M0851) monoclonal antibodies, and anti-protein S-100 (1:4000, Dako Z0311) polyclonal antibody. 2 RESULTS Arising from the striated muscle, there was an un-encapsulated, infiltrative, densely cellular neoplasm composed of thick interlacing bundles. The neoplastic cells were spindle-shaped, 25 micrometers in diameter, or giant and polygonal, up to 100 micrometers in diameter, with abundant eosinophilic cytoplasm and one or two hypochromatic nuclei with prominent nucleoli. Abnormal mitotic figures were present. The neoplastic cells stained positive for vimentin, desmin, and smooth muscle actin, but negative for Protein S-100. The neoplasm was diagnosed as a cervical rhabdomyosarcoma, without evidence of vascular emboli. At the following presentation, 15 days after, the healing was good, the head was in a normal position and the bird was healthy. Two years after surgery, no recurrence was observed. 3 DISCUSSION Reports of neoplasms in birds are numerous, but muscular neoplasms are not frequent (LATIMER 1994, SCHMIDT 1997). Smooth muscle tumours are reported twice as frequently as striated ones (LATIMER 1994). Rhabdomyosarcoma are generally described as irregular, elevated, lobulated and firm subcutaneous swelling of the wing or shoulder (LATIMER 1994). They are frequently immobile and firmly attached to the bone (LATIMER 1994, FERNANDEZ-BELLON et al 2003). In our case, the location and the mobility of the tumour may be considered as infrequent. Most cases are described on dead birds. In hens, rhabdomyosarcomas were reported in the anterior uvea, orbit (DUKES and PETIT 1983), heart, in skeletal muscles (FUJIMOTO and OKADA 1970), breast muscle (with metastasis in liver, lungs and kidney) (ERER and KIRAN 1989) and the biceps femoralis (with spleen metastasis) (FUJIMOTO and OKADA 1970). In a vulture, rhabdomyosarcoma within the myocardium was reported (DUNCAN and FITZGERALD 1997). In a racing pigeon (Columba livia), a 474 rhabdomyosarcoma was described on the ventral aspect of the wing (FERNANDEZBELLON et al 2003). The bird was euthanised and immunohistochemistry was positive for muscle actin and myoglobin and negative for desmin, neuron-specific enolase and S-100 protein (undifferentiated rhabdomyosarcoma) (FERNANDEZBELLON et al 2003). In a budgerigar, a surgical treatment of a rhabdomyosarcoma was not successful. Only 90% of the neoplasm was removed and the bird died 3 hours after surgery (RAPHAEL and NGUYEN 1980). On necropsy, liver metastases were found. Few cases of successful surgical treatments of neoplasms are described. Most of the time, benign tumours are successfully treated (lipoma) (SCHMIDT 1997). Intraabdominal tumours are known to be difficult to remove. In our case, the success of the surgical treatment may be related to the mobility and the location of the mass, and the absence of metastases. The large surgical resection was sufficient. To our knowledge, no successful treatments of rhabdomyosarcoma had been previously reported. 5 CITATION INDEX DUKES TW and PETIT JR. Avian ocular neoplasia-a description of spontaneously occurring cases. Canadian J Comp Medi 1983; 47: 33 - 36. DUNCAN AE and FITZGERALD SD.. Multiple primary rhabdomyosarcomas within the myocardium of a vulture. Clinical challenge 1997; 28(4): 501 - 503. ERER H and KIRAN MM. A case of an undifferentiated rhabdomyosarcoma in a hen. Berl Munch Tierarztl Wochenschr 1989; 102(11):382 - 385. FERNANDEZ-BELLON H et al. Rhabdomyosarcoma in a racing pigeon (Columba livia). Avian Pathol 2003; 32(6): 613 - 616. FUJIMOTO Y, OKADA K. Rhabdomyosarcoma in the chicken. Jap J Vet Res 1970; 18: 109 - 115. LATIMER KS. Oncology. In RITCHIE BW et al, Avian medicine: Principles and Application, 1st ed., Lake worth, FL: Wingers publishing 1994: 640 - 672. RAPHAEL BL and NGUYEN HT. Metastasizing rhabdomyosarcoma in a Budgerigar. J Am Vet Med Assoc 1980; 177(9): 925-926. SCHMIDT RE. Neoplastic diseases. In ALTMAN RB et al. Avian Medicine and Surgery, 1st ed., Philadelphia, PA: WB Saunders 1997:590 - 603. AUTHORS ADDRESS Emmanuel Risi, Centre Vétérinaire de la Faune Sauvage, Ecole Nationale Vétérinaire de Nantes, Atlanpole – La Chantrerie, BP 40706, 44307 Nantes cedex 03, France, Email:
[email protected] 475 Central Veterinary Research Laboratory Dubai, United Arab Emirates ENDO-PARASITES OF THE ASIAN HOUBARA BUSTARD (Chlamydotis undulata macqueenii) R. K. Schuster, U. Wernery, J. Kinne KEYWORDS Chlamydotis undulata macqueenii - Acanthocephalans - Cestodes – Nematodes - U.A.E. ABSTRACT In January 2004 thirty-one carcasses of houbara bustards (Chlamydotis undulata macqueenii) were sent for necropsy to the Central Veterinary Research Laboratory. All birds were heavily infested with endo-parasites. Two species of acanthocephalans (Empodius taeniatus, Sphaerirostris embae, three species of cestodes (Otiotaenia conoideis, Ideogenes otidis, Hispaniolepis falsata) and one nematode species (Harteria rotundata) were identified. In addition, one unidentified Eimeria species was found in a single bird. 1 INTRODUCTION The houbara bustard (Chlamydotis undulata) occurs in 3 subspecies in Asia (C. u. macqueenii), Northern Africa (C. u. undulata) and on the Canary Islands (C. u. fuertaventurae), respectively. The Asian representative is a migratory bird with breeding sites in Central Asia. During the winter these birds migrate south and some of them enter the Arabian Peninsula. C. u. macqueeni is the favourite prey for hunting falcons and an unknown number of these birds are caught in countries north of the Arabian Gulf and smuggled into the U.A.E (BAILEY 2004). The knowledge on parasites of this bustard species is based on a small number of examined birds (WERNERY et al. 2001). In a larger survey in 2003 we examined 104 carcasses of originally wild caught houbara bustards kept at different places in Dubai (SCHUSTER and KINNE 2003). Most of these birds got however, anthelminthic treatment on arrival but drugs used were not indicated. 476 2 MATERIAL AND METHODS In January 2004, thirty-one carcasses of confiscated wild caught houbara bustards were sent to the Central Veterinary Research Laboratory for necropsy. Parasitological examination concentrated on endo-parasites. For this, the intestines were gently opened and helminths were isolated and washed in normal saline. For identification, nematodes and acanthocephalans were cleared up in lactic acid while cestodes were stained in lactic carmine. In addition, a faecal sample taken from the large intestine of each bird was examined using the flotation method. 3 RESULTS All birds were infested with endo-parasites. The helminth spectrum consisted of two acanthocephalans (Empodius taeniatus, Sphaerirostris embae, three cestodes (Otiotaenia conoideis, Ideogenes otidis, Hispaniolepis falsata) and one nematode (Harteria rotundata). Prevalence and burdens of these helminths are given in Tab. 1. The number of birds harbouring one, two, three, four and five worm species was 5, 9, 10, 6 and one, respectively. In addition, a low number of unsporulated oocysts of an unidentified Eimeria species were detected in the faeces of a single bird. Table 1: Prevalence and number of helminths in houbara bustards (n = 31) Parameter species Empodius Sphaerirostris Otiotaenia taeniatus embae conoideis 67.74 5.9 1 39 48.39 53.2 5 315 45.16 14.1 2 125 Ideogenes otidis 19.35 141.3 32 1200 Hispaniolepis Harteria falsata rotundata 32.26 40.5 3 100 58.06 11.8 1 156 prevalence% av. burden min. burden max. burden 4 DISCUSSION All helminths isolated are considered to be bustard specific but some of them might be found in other birds sharing the same environment and having similar food preferences. All isolated worms have a heteroxenic life cycle with arthropods as intermediate hosts. In comparison to the previous investigation (SCHUSTER and KINNE 2003) the current study reflects the helminthological situation in wild houbara bustards more realistically since no anthelminthic treatment was given. All birds harboured at least one parasite species and the prevalence of cestodes and H. rotundata was double high as in the former investigation where most of the birds got an anthelminthic treatment with one of the following compounds: praziquantel, fenbendazole, ivermectin and doramectin. While praziquantel is highly effective against cestodes, the macrocyclic lactones ivermectin and doramectin are administered for nematode infections, and fenbendazole can be used for the control of both cestodes and nematodes. None of the available anthelminthics are acting against acanthocephalans and for this reason the figures for prevalence and abundance of E. taeniatus and S. embae were comparable in both treated and untreated groups. 477 5 CITATION INDEX 1. 2. BAILEY T. Health status of free-living and rehabilitated houbara bustards. Proceedings 3rd Annual Wildlife Disease Association - Africa & Middle East Section, Abu Dhabi, in press. SCHUSTER RK and KINNE J. Acanthocephalans and other helminths of the Asian houbara bustard (Chlamydotis undulata macqueenii). 9th Internat Helminthol Symp, Programme and Abstracts, Stara Lesna, High Tatras 2003: 37. WERNERY U, MOLNAR L and HUNT K. Disease status of wild houbara bustards (Chlamydotis undulata macqueenii). Proc Euro Assoc Avian Vet, Munich 2001: 268 - 269. 3. AUTHORS ADRESS Prof. Dr. R. K. Schuster Central Veterinary Research Laboratory, PO Box 597, Dubai, United Arab Emirates Email:
[email protected] 478 Dubai Falcon Hospital, Dubai, United Arab Emirates ANTIFUNGAL SUSCEPTIBILITY TESTING OF FUNGI ISOLATED FROM THE RESPIRATORY TRACT OF FALCONS C. Silvanose, BSc, DMLT; T. Bailey, MSc, MRCVS, PhD, Dip ECAMS and A. Di Somma, DVM KEYWORDS Aspergillus sp - Minimum inhibitory concentration - Amphotericin B - Itraconazole - Voriconazole ABSTRACT As new anti-fungal agents are introduced for the treatment of fungal infections in birds, establishing minimum inhibitory concentrations (MICs) of fungal isolates from birds is necessary. Biopsy samples were collected during endoscopy from the air sacs of forty-five falcons diagnosed with fungal diseases of the lower respiratory tract. Our data indicated that antifungal tests before and during treatment may be helpful in guiding appropriate antifungal therapy in avian species. 1 INTRODUCTION Aspergillosis is the most common avian mycosis, especially in captive waterfowl, wading birds, penguins, raptors, ostriches, pheasants and passerines (DEEM 2003, POLLOCK 2003, BAUCK 1994, KEARNS 2003, PEREZ 2003, WERNERY 2004). Among raptors, goshawks (Accipiter gentilis) gyr falcons (Falco rusticolus), immature red-tailed hawks (Buteo jamaicensis), golden eagles (Aquila chrysaetos) and snowy owls (Nyctea scandiaca) are more likely to develop the disease (REDIG 2000). Aspergillosis is the most important cause of morbidity and mortality in falcons trained for falconry in the Middle East (SAMOUR 2000). The disease is caused by Aspergillus fumigatus and less commonly by A. flavus and A. niger (BAUCK 1994, KUNKLE 2003, KEARNS 2003). Due to the life threatening nature of these infections and reports of drug resistance, susceptibility testing of pathogenic fungi has become very important in human medicine. As new anti-fungal agents are introduced for the treatment of fungal infections in birds, establishing minimum inhibitory concentrations (MICs) of fungal isolates from birds is necessary. 479 2 MATERIALS AND METHODS This study was carried out at the Dubai Falcon Hospital, from 2002 to 2004. Biopsy samples were collected during endoscopy from the air sacs of forty-five falcons diagnosed with fungal diseases of the lower respiratory tract. Falcon species and numbers included; peregrine (F. peregrinus) - 5, saker (F. cherrug) - 5, gyr (F. rusticolus) – 15, barbary (F. pelegrinoides) -1 and hybrid falcons - 19. hybrid falcons included gyr x peregrine (F. rusticolus x F. peregrinus) - 13, gyr x saker (F. rusticolus x F. cherrug) - 5, and gyr x lanner (F. rusticolus x F. biarmicus) - 1. Forty-five fungal isolates were collected on first presentation before antifungal treatment (Group - 1) was initiated. Fungal isolates were collected during Amphotericin B (AMP) treatment from 8 cases (Group – 2), from 7 cases during Itraconazole (ITZ) treatment (Group – 3) and from 15 cases during Voriconazole (VOC) treatment (Group – 4). Among the treated cases, samples were collected three times from one case during therapy. The antifungal strips used were E strips containing AMP (concentration range, 0.002 to 32µg/ml), ITZ (concentration range, 0.002 to 32µg/ml) and VOC (concentration range, 0.002 to 32µg/ml) supplied by AB Biodisk, Solna, Sweden. The media used for this study was RPMI agar includes RPMI, 1640 with L- glutamine and MOPS buffer. 3 RESULTS AND DISCUSSION An antifungal study was carried out to determine the MIC of fungi isolated from the air sacs of falcons before (Group - 1) and during (Group – 2, 3 & 4) antifungal treatment with amphotericin B (AMP), itraconazole (ITZ) and voriconazole (VOC) respectively. Ninety five percent of isolates (Group - 1) including Aspergillus fumigatus, A. flavus, A. niger and A. terreus were susceptible to VOC at MICs ≤ 0.38 µg/ml and 100% of the isolates at MICs ≤ 1 µg /ml. Twenty one percent of isolates (Group - 1) including A. fumigatus (27.6%), A. flavus (16.6%), A. niger (100%) and A. terreus (23%) had an MIC to ITZ ≥ 1 µg/ml. In human medicine fungi with an MIC ≥ 1 µg/ml are considered resistant. Fifty one percent of isolates (Group - 1) including A. fumigatus (31%), A. flavus (78%), A. niger (14%), and A. terreus (77%) had MICs > 1 µg/ml to AMP. The MICs of AMP for Aspergillus strains during antifungal therapy (MIC ≥ 3 µg/ml, median 8 µg/ml, range 3-32 µg/ml) were significantly higher than the MIC’s before therapy. There was no significant difference between different Aspergillus strains for the MIC of VOC and ITZ before and during antifungal treatment, except one case of A. flavus. In-vitro antifungal tests before and during therapy shows that the majority of Aspergillus sp. are susceptible to VOC at low concentrations. In our study, A. niger was resistant to ITZ but, susceptible to AMP at low concentrations. Isolates showed significant changes of MIC or resistance to AMP, during therapy. Our data indicated that antifungal tests before and during treatment may be helpful in guiding appropriate antifungal therapy in avian species. 480 4 CITATION INDEX 1. BAUCK L. Mycoses. In: BW RITCHIE, GJ HARRISON and LR HARRISON. (eds): Avian Medicine Principles and Application. Lake Worth: Wingers Publishing Inc. 1994; 997 - 1006. DEEM SL. Fungal diseases of birds of prey. Vet Clinics North America: Exotic Animal Practice 2003; 6: 363-76. KUNKLE RA. Fungal infections. In: Y.M. SAIF (ed): Diseases of Poultry, 11th edition. Iowa: Iowa State Press. 2003; 883-895. REDIG P. Fungal diseases: Aspergillosis. In: J. SAMOUR (ed): Avian Medicine. Harcourt Publishers Ltd, London: 2000; 275 - 278. PEREZ J, GARCIA, PM, MENDEZ A, et al. Outbreak of aspergillosis of adult ostriches. Vet Rec 2003; 153: 124 - 125. SAMOUR JH. Veterinary considerations during the hunting trip. In: J.T. LUMEIJ, J.D. REMPLE, P.T. REDIG, et al. (eds): Raptor Biomedicine III. Zoological Education Network, Lake Worth. 2000; 267 - 274. WERNERY R, WERNERY W, KINNE J and SAMOUR J. (eds): Fungal diseases. In: Colour Atlas of Falcon Medicine. Hanover: Schluetersche 2004; 81 – 91. 2. 3. 4. 5. 6. 7. 5 ACKNOWLEDGEMENTS We thank HH Sheikh Hamdan bin Rashid al Maktoum, Deputy Ruler of Dubai, Finance and Industry Minister of UAE for his continued support of the Dubai Falcon Hospital. We thank Mr Humaid Obaid al Muhari, the staff of the Dubai Falcon Hospital and Mr. Peter McKinney (Falcon Center, Dubai) for their technical support. AUTHORS ADRESS C. Silvanose Dubai Falcon Hospital, P.O.Box 23919, Dubai, United Arab Emirates Email:
[email protected] 481 482 . Tena Pera. blood samples (for haematology. blood extraction (from the external cubital vein) and collection of a faeceal sample. The health of captured individuals was assessed through a comprehensive clinical examination. Animals caught for radiotracking projects were individually checked for health problems and a parasitic survey was conducted at several lek places. A 33% ZnSO4 flotation technique was used to give qualitative results and the McMaster technique was used for the quantitative results. DVM. Fresh faecal samples were collected from several lek sites. KEYWORDS Analysis .Les Valls Veterinary Hospital. 2 MATERIAL AND METHODS Capture: Animals were captured using nylon nets on the leks.Samples . biochemistry and serology for selected infectious diseases).Capture . A complete clinical inspection was performed: external inspection. The health survey was used to complete the demographic data and to find new indicators useful for management decisions. ectoparasites and endoparasite.Capercaillies ABSTRACT In the principality of Andorra (Central Pyrenees). 1 INTRODUCTION The demographic survey has enabled collection of numerous data about the general tendency of the Andorran capercaillie’s population . The framework of health control includes capture operations and clinical analysis. A parallel parasitological study was made by faecal collection in the birds’ singing sites covering the entire country. the capercailles are included in the list of protected species. Principality of Andorra HEALTH CONTROL OF VARIOUS CAPERCAILLIES (TETRAO UROGALLUS) CAPTURED IN THE PRINCIPALITY OF ANDORRA J. Physical restraint was used. Internal parasites: see section IV 3. 2 AD 500 Andorra la Vella. 3. The liver parameters were within normal range with the exception of one individual. No individuals have died during or post-capture. capercaillie density might have had an impact on the presence of parasites. It is possible that human presence. Haematological parameters are within normal range. Depending on the previous factors cited. 5 CITATION INDEX References available from the author AUTHORS ADRESS J. optimum leks could be identified. vehicles. In terms of the parasitology results we consider it likely that a stressor (human presence.3 RESULTS 3. The data obtained on captured animals has not identified major sanitary problems. paramyxovirus and influenza virus tests were all negative.2 Parasitic study External parasites (ticks and lice): no parasites were observed. Higher muscular markers (CPK.1 Results on captured animals: a total of 80 % of the animals had a Body Condition (BC) of 4. 4 DISCUSSION The number of individuals that have been studied is low. Tena DVM Les Valls Veterinary Hospital C/Joan Maragall. GORTAZAR 2000). LDH) than those established for the Gallus domesticus are probably due to stress related to capture or to the mating season (LIUKKONEN-ANTTILA 2000). 3. Email:
[email protected] 483 . 20% of the animals had a BC of 3. capercaillie density) has an impact in the presence of parasites (BELLEAU 1991. further capture and marking tasks will produce further data. vehicles. Principality of Andorra.3 Infectious diseases serology Chlamydophila.4 – The parasitological survey A 70% frequency of coccidia and a 100% of cestoda was observed in the leks located less than 100 metres of a road. No lesions or injuries were found on external examination. Allocating thirty minutes to handling enabled marking of the animal and collection of thirty-seven physical and clinical variables. JCCM Instituto de Investigación en Recursos Cinegéticos IREC Spain TREATMENT OF CROP FISTULAS DUE TO SEVERE TRICHOMONIASIS IN TWO BOOTED EAGLES (HIERAAETUS PENNATUS) S.Trichomoniasis ABSTRACT Trichomonas gallinae may provoke the development of large fibronecrotic lesions in the upper digestive tract and occasionally other locations of columbiforms and other avian species. U. but also passerines and psittacine species. These lesions generally lead to death of the affected bird from starvation or secondary infection. Blanco DVM KEYWORDS Booted eagles . especially in pigeons the parasite is frequently present in absence of clinical lesions. ESVA. fibronecrotic lesions that develop in clinical disease may produce impairment of the normal motility of the crop and oesophagus due to the partial obstruction of the crop lumen and destruction of the mucosa and adjacent tissue. Höfle DVM.M. Asistencia Veterinaria sl Centro de Estudios de Rapaces Ibéricas CERI. Both animals had also intestinal parasites and one of them had an infection with Candida albicans.Hieraaetus pennatus . Villaverde. Trichomoniasis related lesions are usually localized in the oropharynx and crop and may occasionally involve the respiratory tract or other locations such as the infraorbital or supraorbital regions (SAMOUR 2000). gallinae from morphology and culture and PCR.Crop fistula . J.Aquila Foundation. Numerous flagellate protozoa were observed on direct examination and identified as T. Treatment was medical and also involved surgery achieving satisfactory healing and recovery in both cases. Although. 1 INTRODUCTION Trichomoniasis is a disease of columbiforms and birds of prey. 484 . Two booted eagles were admitted to the Centro de Estudios de Rapaces Ibéricas with large fibronecrotic lesions and associated fistulas in the crop. It is caused by the flagellated protozoan Trichomonas gallinae. In both booted eagles other pathogens were found. Hydration was maintained with intravenous fluids (Ringer’s lactate). Both birds were severely emaciated upon admission and the male presented an additional firm. In the female C. In the other case a one-layer continuous inverted suture with 4-0 Vicryl was used and an additional dressing to protect the wound was applied. and were identified as T. The male was also positive for the presence of Capillaria and suffered from a low infestation with Leucozytozoon. Candidiasis was treated using oral 485 . a male and a female were admitted to the Centre for Studies on Iberian raptors (CERI) with well developed fibronecrotic lesions in the crop that had led to external fistulae in both cases. relative and absolute eosinophilia and marked toxicity of the heterophils (Grade II-IV). Both birds were re-cultured twice for the presence of T. albicans was cultured from the lesion and propagules of Capillaria and were observed on faecal flotation (344 propagules/g of faeces) while on direct examination of faecal matter. evident on the blood smear. The eagles were received within one month but originated from different regions. In addition blood samples were obtained from both birds and total white cell counts. rounded lesion of about 4mm in diameter on its left eyelid. and electrophoresis of plasma proteins were performed.2 MATERIAL AND METHODS Two juvenile booted eagles (Hieraaetus pennatus). This fistula required continuous cleaning and refreshing during 5 days before excision of the full extent of necrotic tissue and suturing was possible. tapeworm propagules were observed. In one of the eagles. In both cases the fibronecrotic mass adhered to the lesion required surgical removal 7-9 days after initiation of medical treatment. cultures for Salmonella and other pathogenic Enterobacteriaceae were carried out from the cloaca as well as faecal flotation for parasite ova. From both birds swabs were taken from the lesions for direct wet mounts and culture for Trichomonas sp. Additional postoperative care included soft diet and antibiotics for 7-10 days after healing of the lesions. the tissue damage and necrosis involved the crop mucosa and skin in a region of approximately 2cm around the fistula.. 3 RESULTS High numbers of flagellated protozoa were observed directly and on culture from the crop lesions and also from the mass on the eyelid of the male booted eagle. We used hydrocolloidal pads for this dressing to absorb fluids and exudates and to encourage granulation and re-epithelialisation. Both eagles had a moderate to marked leucocytosis. Both cases were treated with a combined therapy using metronidazole (local) and carnidazole (15mg/kg single dosis). differential white cell counts. Sistemic antibiotic treatment (Piperacillin 20mg/kg IM BID) and clorhexidine flushing of the crop were used to control secondary infections due to the presence of necrotic tissue. pathogenic fungi and bacteria. albumin/globulin rates were decreased due to mainly a marked decrease in albumin and an increase in the alpha 2 and beta fractions. gallinae by morphology and PCR. gallinae throughout a period of two months after surgery and treatment. Also. Upon plasma protein electrophoresis. Closure of the surgery site was obtained using a two-layer suture in one of the cases. both with 4-0 Vicryl. the first layer with a continuous inverted pattern and the second layer using a simple interrupted suture. S. regaining full normal motility of the oesophagus and crop in absence of any constrictions due to scar tissue. Trichomoniasis in a Bonelli’s eagle population in Spain. REMPLE. Trichomoniasis in free-living goshawks (Accipiter gentilis gentilis) from Great Britain. BLANCO J M. while in others. J Wildl Dis 2000. In: LUMEIJ. et al. KS. J Wild Dis 1998.52. BOAL CW. J. 4 DISCUSSION T.T. C. Trichomoniasis in Bonelli’s eagle (Hieraaetus fasciatus) nestlings in South-west Portugal. 24: 80 . Trichomoniasis in Cooper’s hawks from Arizona. gallinae in the subsequent analysis. The association of trichomoniasis with infection by C. 4. COLE R A. Direct microscopy would have been sufficient for diagnosis of trichomoniasis in this case. Healing of the lesions and function of the crop was satisfactory in both eagles. 45 . 2000). SAMOUR JH. PALMA L and MELO P. Both birds were released back into the wild about six months after admission. COOPER J E and PETTY S J. albicans is consistent with previous observations in Bonelli’s eagle (Hieraaaetus fasciatus) nestlings (HÖFLE et al. culture has proven more effective (KOCAN and KNISLEY 1970). candidiasis and parasitosis. 5 CITATION INDEX 1. gallinae is generally considered a secondary pathogen.140. and FRANSON. D.. MAÑOSA S and MUÑOZ E. (eds): Raptor Biomedicine III. After treatment none of the two birds cultured positive for T. J. Summer in central Spain is very dry and hot. P. M. 2.206. This and the fact that these juveniles have recently become independent from their parents may have caused them to prey on diseased pigeons or feed on carrion and also rendered them more susceptible to the disease. 486 .593. (eds): Field Manual of Wildlife Diseases. and both birds recovered fully with treatment of trichomoniasis. 36: 64 . REAL J. MANNAN RW and HUDELSON. Lake Worth: Zoological Education Network. National Geological Survey.. 5. Supraorbital trichomoniasis infection in two saker falcons (Falco cherrug) Vet Rec 2000. In the booted eagles in this case no other pathogen or primary traumatic lesion could be detected.87. and food and water resources are scarce. 2000). Department of Interior. REAL et al. Trichomonosis. In: FRIEND. 2000. Inc 2000. Potential subjacent viral infections however were not analysed in this case. However the disease has been considered an emerging disease in free-living nestlings of birds of prey in Spain in relation to changes in prey availability (HÖFLE et al.Itraconazole (15mg/kg) and the helminthosis resolved with oral levamisole (4mg/kg) and praziquantel. Both eagles fully recovered their physical condition.70. HÖFLE U. J. REDIG. U. especially in columbiforms were prevalence of the parasite is high and only occasionally associated with disease (KOCAN and KNISLEY 1970). J Wildl Dis 1988.T. 146: 139 . especially in subclinical infections. 34: 590 . 6. 201 . 3. 1999. 45671 Toledo.R. Challenge infection as a means of determining the rate of disease resistant Trichomonas gallinae free birds in a population. C. 135: 358-362. Sevilleja de la Jara. BAILEY TA and COOPER JE.es 487 .15. Vet Rec 1995.E. 6: 13 . AUTHORSS ADRESS S. 8. Asistencia Veterinaria s.I.l.. Centro de Estudios de Rapaces Ibéricas. Villaverde ESVA. Spain Email: cerito@jccm. SAMOUR JH. Trichomoniasis in birds of prey (order Falconiformes) in Bahrain.7. J Wildl Dis 1970. KOCAN RM and KNISLEY JO. caudal). Air enters the nasal cavities via the nares. The nasal cavities are separated by an osseous septum (vomer bone). The caudal (or olfactive) concha is not present in the African grey parrot (Psittacus erithacus) (KRAUTWALD-JUNGHANS et al. 1998). preauditory. Various treatments and laboratory tests were started over several months without success. It will circulate in the IOS. France MUCOCELE IN A LESSER SULFUR-CRESTED COCKATOO (CACATUA SULPHUREA) J. Both side possess nares. Finally a biopsy revealed a mucocele.5 to 3 mm in diameter) into the nasal cavity to allow drainage of secretions. to be moistened. KEYWORDS Parrot – Mucocele ABSTRACT A 9 year-old lesser sulphur-crested cockatoo (Cacatua sulphurea) was presented with a 3-month history of chronic sinusitis that was non-responsive to treatment. middle. Usually. The nares are the inlet of nasal cavities. Surgery alone resolved that apparently unusual case. warmed and filtered in the conchae. cervico-cephalic air sacs and choanae (ANDRÉ 2004).P. making draining of a purulent secretion difficult. Degorce-Rubiales. Secretions of the lacrymal glands are drained into the nasal cavity (middle concha) via lacrimal ducti.Laboratoire d’Anatomie Pathologique Vétérinaire du Sud-ouest. maxillary). DVM. infra-orbital sinuses (IOS). The IOS consists of several diverticulae (infra-orbital. In parrots the right and left IOS are in communication. The last concha does not communicate directly with the nasal cavity. André. Each IOS has a very small dorsal opening of the medial concha (0. DVM. each nasal cavity is divided into 3 conchae (rostral. cervico-cephalic air sacs and cranial pneumatic bones before entering the oropharynx via the choana. mandibular) and 2 chambers (suborbital. Toulouse. 1 INTRODUCTION The upper respiratory system of birds is very complex. 488 . rostral. and F. pre-orbital. The middle concha communicates with the buccal cavity by means of the choana (“internal nares”). nasal cavities. it was alert and weighed 270g. Under a keratinized Malpighian epithelial tissue. “Jonquille” seemed in good body condition. without recurrence of swollen IOS. close by these deposits of tissue. one could consider that it is a complication of a chronic sinusitis. oedematous tissue that was very difficult to clear. often surrounded with a strip of macrophagic inflammatory cells. reminiscent of a sialocele or mucocele. many sinus washings with daily cleaning of nares were carried out. The diagnosis was confirmed by G. BELISA and ELISA tests for Chlamydophila psittaci were negative. The mucus produced by the glands of the hyperplasic respiratory epithelium could account for the retention in connection with a lack of sinus drainage 489 . Neighbouring feathers were moist and matted. “Jonquille” has been in good health. Therefore. very few nonimpaired white cells. All treatment was stopped 10 days before our examination. The bird was head-shaking as if to eliminate this discharge. with or without anaesthesia. Conclusion: a sialocoele (or mucocele). intrusion of which gives rise to a granulomatous inflammation. No diagnostic tests had been performed. or an inflammatory cyst extended around the secretory product of a mucous gland”. cystic lesion of the sinuses which is lined by a pseudostratified or columnar epithelium (PRATER 1997). numerous “puddles” of a mucinous bluish product can be observed. Since then. “Jonquille” lived in a small inside aviary. Several surgical operations were still necessary to eliminate the abnormal tissue. On physical examination. Cytology showed desquamation cells. fruits and vegetables. 3 DISCUSSION The description of a mucocele in a bird appears to be quite unusual. She had laid one egg 4 years before. one observes. In birds. one observes glandular structures (more exactly tubular structures) covered with a double layer of epithelial cells reminiscent of salivary or mucous glands excretory ducts. The chief complaint was a chronic sinusitis for 3 months. but no bacteria nor yeasts. During a 5 month period. slimy.2 CLINICAL REPORT A 9-year old female lesser sulphur crested cockatoo named “Jonquille” was presented to our veterinary office. Veterinary Faculty of Utrecht (personal communication). A week later. The laboratory report was as follows: “Several samples were examined. At times. histologic appearance is in favour of a breaking or trauma of these excretory structures that accounts for the intrusion of the material secreted in the surrounding chorion. The left nare was mis-shapen (oval). To investigate a possible myxoma. surgery was undertaken on the left side to remove gelatinous material. but the sinusitis had not resolved. She was fed seed. A significant translucent nasal discharge was associated with this. in the underlying fibrous chorion. Cultures were negative for bacteria and fungi. Its face was very swollen all around the eyes. very likely derived from a mucous membrane. Dorrestein. In man it concerns a chronic. Various antibiotics and aerosols had been prescribed by their usual veterinarian. surgery was performed on the right side and a sample of the gelatinous material was taken for histologic analysis. A bilateral incision of the infra-orbital diverticulum of the IOS revealed a translucent. ANDRÉ JP. 27bis rue du 14 juillet. (ed) MIPP-F. F. UTMB. 1-8.33130 Bègles 2004. AUTHORS ADDRESS J. March 1997. Oiseaux de cages et de volières. André. De la maladie à la bonne santé. of Otolaryngology.157. P. ANDRÉ JP. the delayed undertaking of a histologic examination. In this clinical report. PRATER ME.(obstruction of the ostia).htm 3.fr 490 . KRAUTWALD-JUNGHANS ME et al. France email: jeanpierre. http://www. 4 CITATION INDEX 1. 2. J Avian Med Surg 1998. Dept. 12: 149 . the duration of disease (8 months) and the difficulties encountered for the removal of all the material that completely moulded the IOS diverticulum. DVM -Cabinet Vétérinaire. Comparative studies on the diagnostic value of conventional radiography and computed tomography in evaluating the heads of psittacine and raptorial birds.33260 La Teste de Buch. it must be pointed out.edu/otoref/Grnds/Sinusitisacute-9703/sinus-acute-9703. Grand rounds presentations. the diagnostic error at the outset as an infectious IOS sinusitis was considered.andre9@free. Acute sinusitis. reaching 6.0/5. Pittman CVT.Johannesburg Zoo South Africa ZINC TOXICOSIS IN A WATTLED CRANE (BUGERANUS CARUNCULATUS) M. Over the six month period. with lower levels in the subsequent cases. her weight decreased further to 5.4kg although she eventually regained the weight lost. The crane was treated medically with chelation agents and had her ventriculus flushed under anaesthesia. 1 CLINICAL REPORT A four year old female wattled crane was treated twice over a period of six months for ingestion of heavy metal. Even though the keepers had checked the enclosure with a metal detector before placing the birds in the new exhibit. M. which the bird and her mate had been moved to a few weeks prior to the first episode. The majority of the metal pieces were galvanised zinc hog rings used in construction of the fencing for a new enclosure.Zinc ABSTRACT This case report details the diagnosis and treatment of zinc toxicosis in an adult female wattled crane (Bugeranus carunculatus). J.Wattled crane .7kg after successful treatment. A serum zinc result of 53umol/l (346ug/dl) was obtained initially. to a maximum of 40. there were no abnormalities on physical exam. The report covers a period of several months during which the crane ingested galvanised zinc hog rings used in construction of a new exhibit on two separate occasions. On each occasion the main clinical signs were weight loss in spite of a normal appetite and a depressed attitude.Toxicosis . She weighed 6. KEYWORDS Birds . Biochemistry results were consistently normal. whereas the white cell count was often but variably raised. Hartley BVetMed MapplSc (Wildlife Health) MRCVS. Barrows BSc BVMS CertZooMed MRCVS. The initial serum zinc level was 53umol/l and the main presenting sign was weight loss. 491 . the female had managed to find and ingest several hog rings and other pieces of metal discarded by the builders. Initially the crane was examined as part of a routine health check and was found to be in poor body condition with a prominent sternum and a body condition index of 2.3kg and apart from the weight loss. M. Interestingly the male bird never ingested any metal.68 x 109/l at the time of initial diagnosis. however approximately one month after the final flushing procedure the crane started to gain weight and was discharged back to her enclosure weighing 5. Blood lead was 4. which had again been searched with a metal detector for any further pieces of metal while she had been hospitalised. raw egg. The decision was then made to flush the ventriculus under anaesthesia and this procedure was repeated on the subsequent occasion. In spite of this she continued to find and ingest the hog rings and so after this a three metre strip of turf was removed from the perimeter of the enclosure before replacing the crane. Post-operative care included clavulanate potentiated amoxycillin at 150mg/kg SC sid. Recovery was complicated by a tracheitis necessitating further diagnostic procedures and treatments.1 mg/kg PO sid and a course of CaEDTA at 32mg/kg IM sid. Water at a temperature of approximately 40 °C was flushed through the tube into the ventriculus and the bird’s head held down over a bucket containing a sieve. Vetark. Analgesics. There has been no recurrence of metal ingestion since. UK). Each time lateral and ventrodorsal radiographs taken under isoflurane anaesthesia revealed several pieces of metal present in the ventriculus. On the first occasion eighteen pieces of metal were removed and on the second occasion twenty pieces. Approximately 50ml/kg warmed Lactated Ringer’s Solution was given intravenously via a catheter in the medial metatarsal vein during the procedure. On the first occasion she was also given cisapride at 1mg/kg PO sid and on the second occasion. meloxicam at 0. after seven days the edetate injections were replaced by 300mg penicillamine given PO for two weeks. antibiotics and a course of CaEDTA were started and continued postoperatively as detailed below. A piece of gauze was held over the choana during the procedure.8 kg. an avian probiotic (Avipro. Four months later her weight had increased to 6.5mm endotracheal tube.Zinc levels greater than 200ug/dl are suggestive of zinc toxicosis (LABONDE 1995). Vetark. Five days later the crane was anaesthetised for repeat radiographs which showed some smaller pieces of wire passing through the intestines but all of the hog rings still present in the ventriculus.0ug/dl. During the first episode the crane was hospitalised and treatment initiated with 200mg (32mg/kg) sodium calcium edetate (CaEDTA) diluted in sterile water and given intramuscularly into alternate pectoral muscles once daily. The second episode occurred after the crane had been discharged back to her original enclosure. a lubricated piece of plastic tubing. 2m long was passed into the ventriculus.7 kg. The bird was kept in ventral recumbency with the neck streteched out and the body inclined at an angle of approximately 60 degrees. 492 . No other abnormalities were seen on the radiographs. Uk) and a calcium/vitamin D3 supplement (ZolCal D. The crane was subdued and anorexic for a day or two after the flushing procedures and so was tube fed with a mixture made from ground trout pellets. a maltodextrin and protein concentrate solution (Critical Care Formula. The crane was fasted overnight and then after anaesthetising and intubating her with a 6. Vetark UK) until her appetite returned. Saunders Co 1997. They then clumped together and got stuck at the base of the neck where repeated flushing attempts were needed to dislodge them. Stainless steel should also be used instead of galvanised zinc.31 493 . 1994.2 DISCUSSION Cranes are particularly susceptible to heavy metal toxicosis due to their propensity to ingest foreign bodies (OLSEN and CARPENTER 1997). Veterinary Clinics of North America: Small Animal Practice. studies on birds have shown a significant diurnal variation in blood zinc concentration (ROSENTHAL et al. On one occasion the pieces of metal were flushed quickly out of the ventriculus into the proventriculus requiring an adjustment in tube position before they could be moved up the oesophagus. LABONDE J. DORRESTEIN GM and QUESENBERRY K (eds): Avian Medicine and Surgery. Seminars in Exotic Pet Medicine. weight loss. CaEDTA is excreted by the kidneys and may be contraindicated in birds with renal compromise. Avian Toxicology. 21(6): 1329 . 3. 3 CITATION INDEX 1. LABONDE J. 1991. It is recommended that liver and renal functions are monitored during chelation therapy. diarrhoea. CaEDTA nephrotoxicity is recognised in humans. 2. Toxic Diseases.B. This case highlights the importance of builders and maintenance workers clearing up meticulously after constructing new exhibits. In spite of attempting to block the choana during the flushing procedure. polyuria. Opting for the gizzard flushing procedure at an early stage is likely to be preferable to lengthy courses of chelation therapy and associated handling.613. MAUTINO 1997). Organs affected by zinc toxicity include the pancreas. 4: 23 . However there are no reliable reports of nephrotoxicity in birds as it has been used for up to 23 consecutive days with no ill effects (LABONDE 1991. The weight loss in spite of a good appetite seen in some cases of zinc toxicity could at least partly be attributed to maldigestion due to pancreatic insufficiency (ZDZIARSKI et al. BAUCK L and LABONDE J. including lethargy. regurgitation. ZDZIARSKI et al. In: ALTMAN RB.1342. liver. depression. BAUCK and LABONDE 1997. as in humans. on every occasion at least one or two pieces of metal or grit from the ventriculus had to be removed from the choana at the end of the procedure. The gizzard flushing procedure was found to be straightforward. 1995. 1994). CLUBB SL. although repeat radiographs were essential to monitor progress. It is important to radiograph the head and neck at the end of the procedure to ensure that there are no pieces of metal left in the oesophagus or oral cavity. Zinc is a cumulative toxin and ingestion can result in a variety of clinical signs in birds. kidney and gastrointestinal tract. Treatment and diagnosis can be complicated by the stress of hospitalisation and handling in these sensitive birds. 2004). Toxicity in pet avian patients. It is important to note when relying on serum zinc levels to diagnose toxicity that. ileus and ataxia (LABONDE 1995. Philadelphia: W. feather picking. 604 . SAMOUR and NALDO 2002). 6. 973 . J Zoo Wild Med 1997. 5. Cranes. Diagnosis and therapeutic management of lead toxicosis in falcons in Saudi Arabia.35. AUTHORS ADDRESS M. CLUBB SL.org.20. BSc BVMS Cert ZooMed MRCVS Professional Zoolife Consultants. DORRESTEIN GM and QUESENBERRY K (eds): Avian Medicine and Surgery. MAUTINO M.445. South Africa Email: Vet@jhbzoo. MATTIX R. Philadelphia: WB Saunders Co 1997. Houghton 2041. JOHMSTON M. Barrows. ZDZIARSKI J. ROSENTHAL K. SHOFER F and POPPENGA R. Lead and zinc intoxication in zoological medicine: a review. BUSH M and MONTALI R. 25 (3): 438 . SAMOUR J and NALDO J. 28(1): 28 . J Avian Med Surg 2002. Heavy metal plasma concentration: daily fluctuations and clinical implications.991. 8.4. PO Box 956. In: ALTMAN RB. J Zoo Wild Med 1994. Proc Assoc Avian Vet.35 OLSEN G and CARPENTER J. 16(1): 16 .za 494 . Zinc toxicosis in diving ducks. Johannesburg. 7. New Orleans 2004: 33 . SCHMIDT RE and FUDGE 1990). Most methods for obtaining such samples rely on removing a feather. BAUCK 1997. There are many documented causes of these syndromes and the majority are not directly related to the skin (COOPER and HARRISON 1994. Diagnosis relies on the cytological examination of feather pulp. and cytology (COOPER and HARRISON 1994. SCHMIDT RE and FUDGE 1990). WELLE 1999). a leucocytosis with toxic heterophilia may also be seen (REAVILL. WELLE 1999. In addition. BAUCK 1997. BAUCK 1997. SCHMIDT RE and FUDGE 1990). FORBES and LAWTON 1996.Feather plucking .Parrots ABSTRACT Feather pulpitis appears to be a common cause or complicating factor in feather plucking parrots. biopsy (COOPER and HARRISON 1994. WELLE 1999). FORBES and LAWTON 1996.R. FORBES and LAWTON 1996.Feather pulp . GILL 2001.Strathmore Veterinary Clinic. FORBES and LAWTON 1996. REAVILL. FORBES and LAWTON 1996. Bacterial infections of the feather pulp are described in the texts but are often described as rare (COOPER and HARRISON 1994. 495 . WELLE 1999). Of these methods. BAUCK 1997.Chitty BVetMed CertZooMed MRCVS KEYWORDS Cytology . 1 INTRODUCTION Feather plucking or chewing are common reasons for psittacine birds being presented to the veterinarian and this is reflected by the literature coverage (COOPER and HARRISON 1994. SCHMIDT RE and FUDGE 1990). FORBES and LAWTON 1996. United Kingdom A NEW METHOD FOR THE CYTOLOGICAL SAMPLING OF FEATHER PULP J. GILL 2001. However these samples are often contaminated with blood from the follicle as the feather is removed. BAUCK 1997. It is also difficult (and painful for the bird) to remove remiges and/or rectrices that may need to be sampled. These syndromes are seen much less commonly in raptors (MALLEY and WHITBREAD 1996).Pulpitis . GILL 2001. Methods used to investigate these infections include culture/ sensitivity (COOPER and HARRISON 1994. WELLE 1999. This method described fine needle aspiration of feather pulp via the superior umbilicus without removal of the entire feather. BAUCK 1997. REAVILL. Andover. REAVILL. COOPER JE and HARRISON GJ. However. FORBES NA and LAWTON MPC. Shurdington: BSAVA: 1996: 211 . 5.221 .124. Veterinary Clinics of North America: Exotic Animal Practice 2001.5ml syringe and a small aspirate obtained. these techniques require feathers to be removed from the bird and. The superior umbilicus of the blood feather to be sampled is identified and the area surrounding it cleaned using chlorhexidine on a cotton-tipped swab. there is difficulty in obtaining a representative sample for the pulp without contamination from blood or from the follicle. Clinical Approach to Feather Picking. Philadelphia: WB Saunders: 1997: 548 – 562. 2 METHOD 1. 4: 463 – 492. Suction is applied using a 2. 4. WELLE 1995) while others only recommend it where there is obvious inflammation or crusting (BAUCK 1997.639. non-invasive. Avian Dermatology. (eds): Avian Medicine: Principles and Application. WELLE K. Sampling in this manner does not appear to increase the chances of the bird damaging or removing the sampled feather although that is hard to determine in a feather damaging bird. In: RITCHIE BW. 4 CITATION INDEX 1. In: Manual of Psittacine Birds. This may provide an indication that innervation of the growing feather is significant and that inflammatory conditions within the pulp are capable of causing discomfort or pain. birds often vocalise when the sample is being aspirated. 4. sensitive and results are obtained quickly. Miscellaneous. In: ALTMAN RB. The bird should be adequately restrained. in the case of body feathers.cytology is simple. Smears are made in the usual manner prior to staining. 496 . A hypodermic needle is passed via the superior umbilicus and into the pulp. HARR 2000). JH. Avian Skin Diseases. Proc Assoc Avian Vet. 3.6 x 30mm needle for remiges/rectrices. the following technique has been developed. Phoenix 1999: 119 . GILL.5 x 16mm needle for body feathers and a 0. HARRISON GJ and HARRISON LR. 3. cellular harvest seems reasonable. The author uses a 0. Techniques for sampling feather pulp are described by ECHOLS (2000) and CHITTY (2002). (eds): Avian Medicine and Surgery. DORRESTEIN GM and QUESENBERRY K. CLUBB SL. 2. Lake Worth: Wingers Publishing Inc 1994: 607 . 2. Dermatology. Some authors recommend cytological investigation of all feather plucking/ chewing cases FORBES NA and LAWTON. Interestingly. 3 DISCUSSION Using this technique. BAUCK L. Therefore. (eds): Manual of Raptors. United Kingdom Email xmo32@dial. HARR KE.com 497 . Clinical Cytology (Avian Specialty Advanced Program) Assoc Avian Vet. Portland 2000: 21 – 26. Clinical Cytology (Avian Specialty Advanced Program) Assoc Avian Vet. SCHMIDT RE and FUDGE AM. Pigeons and Waterfowl. REAVILL DR. Sample Collection. ECHOLS S. The Integument. Avian Integument. Portland 2000: 3 – 16. New Orleans 1990: 248 – 255. 7. 8. Cytological Sampling in Avian Skin Disease.6. AUTHORS ADDRESS JR Chitty BVetMed CertZooMed MRCVS Strathmore Veterinary Clinic. Preparation and Handling.pipex. Avian Skin and Feather Disorders: A retrospective study. London Road. Monterey 2002: 355 – 358. 1996: 129 . MALLEY AD and WHITBREAD TJ. In: BEYNON PH. FORBES NA and HARCOURT-BROWN NH. Shurdington: BSAVA. Proc Assoc Avian Vet. Andover. 10. CHITTY JR. Hants SP10 2PH.139. 9. Proc Assoc Avian Vet. both in healthy and in sick birds.Klebsiella oxytoca .Respiratory infection ABSTRACT Klebsiella species was isolated from healthy and from birds with upper respiratory tract infection considered to be potential pathogens of the respiratory tract (primary or opportunities). Nasopharyngeal infection resolved completely in 12 days. inflamed nasal and pharyngeal mucosa with lot of mucopurulent discharge and plugged right nare. with correct feeding and without stress. E. Prukner-Radov�i� KEYWORDS Monk parakeet. The presence of K. Horvatek. in which they can act as primary pathogens or can be involved as opportunists in immunosupressed or stressed birds (GERLACH. under certain circumstances. I. Treatment included enrofloxacin orally every 12 ours for 10 days.. coughing. Ciglar Grozdani�. Gottstein. 498 . JESUS (1998) reported K. oxytoca. Croatia KLEBSIELLA OXYTOCA INFECTION IN MONK PARAKEET D. present in intestinal tract of man and animals (Kelly et al. oxytoca and Klebsiella pneumoniae to be potential pathogens of the respiratory tract. non motile. without any contact with birds. �. H.Faculty of Veterinary Medicine University of Zagreb. A 17-year-old female monk parakeet (Myiopsitta monachus) came with multiple signs of respiratory distress and loss of weight. 1985). Abnormalities shown on physical examination included open-mouthed breathing with severe dyspnoea. oxytoca in bird housed in optimal condition. both in healthy and in sick birds under stress or immunosupression. 1 INTRODUCTION Klebsiella oxytoca is encapsulated. Cultures from pharynx and nares revealed heavy growth of Klebsiella oxytoca and Staphylococcus spp. Klebsiella species are mainly isolated from finches. was unexpected finding. facultative anaerobic Gram-negative bacteria. Mazija. 1994). COLES (2001) reports about an outbreak of juvenile bird mortality because of pollution of drinking water supply with K. given by the owner under prescription and supervision. Biorad. Pliva. and was maintained on seed diet with lot of different fruits and vegetables every day. oxytoca. Swabs were placed directly on Nutrient Agar (Becton Dickinson. like symptomatic therapy. Gram staining. Cockeysville. France). Biorad. The bird had no previous history of any medical problems. Biorad. strictly under our instruction and supervision. Zagreb. Lyon. most of its time in the cage without any contact with other birds.2 CASE REPORT Bird was kept alone in a big stainless-steel cage. oxytoca revealed. and biochemical characteristic using API 20 E strips (BioMerieux. without performing any other examination. Biorad. the bird was physically examined daily. first dose of enrofloxacin (5mg/kg IM q12 h. sulfonamides (200µg. The symptoms worsened despite the therapy. France) and resistant to clarithromycine (15µg. Prior to releasing from clinic. Abnormalities included openmouthed breathing with severe dyspnoea. in the living room of owners’ apartment.5%. every 12 ours in peace of fruit or cookie. An initial workup preformed included bacteriological and mycological examination of swabs taken from pharynx and nares. Mycological examination was negative. weight loss and normal faecal discharge. coughing. Croatia) given by the owner (the patient was managed as outpatient) for 10 days. Biorad. Cockeysville. The antimicrobial susceptibility test (Bauer-Kirby test) performed for K. Owner reported the bird hasn’t eaten for the last 3 days. oxytoca and Staphylococcus spp. France) and sulphonamides (200µg. indicated by the zone of inhibition. France). USA). All clinical signs resolved with treatment completely after 12 days. but he prescribed sulfonamides and vitamins. 499 . in spite of good housing. France). During the therapy. aerobically. Physical examination revealed poor body condition. A faecal Gram stain was normal. inflamed nasal and pharyngeal mucosa with lot of mucopurulent nasal discharge and plugged right nare. USA). The taxonomic classification of isolated bacteria was based on colony morphology. oxytoca was very sensitive to enrofloxacin (5µg. had respiratory difficulties for two weeks (intensive cough. France). Bayer. Biorad. Vetoflok 10%. Germany) and Sabouraud’s Dextrose Agar (Becton Dickinson. Germany) and multivitamins was administered intramusculary. Prior to our examination. Germany). Cultures from pharynx and nares revealed heavy growth of K. that K. semi-sensitive to cefotaxime (30 µg. nasal discharge). Fourteen days after complete recovery. France). Bayer. amoxycillin/clavulanic acid (20/10µg. tetracycline (30µg. BPLS-Agar (Merck. Treatment included enrofloxacin (30mg/kg p/o q12h. and Ringers solutiosubcutaneously. Baytril 2. Biorad. choanal and pharyngeal swabs were taken and cultures revealed no presence of K. another veterinarian had seen the bird and found nothing significant. France). ampicillin/sulbactam (10/10µg. Darmstadt. 309. The bird had no previous history of medical problems.10 000 Zagreb. Proc IVCVM: Diseases of Psittacine Birds 1998 (www.3 CONCLUSION A 17-year-old female monk parakeet (Myiopsitta monachus) came with multiple signs of respiratory distress and weight loss. First therapy (sulphonamides) given by another veterinarian didn’t results with recovery. kept most of its time in the cage and was maintained on seed diet with lot of different fruits and vegetables every day. Saunders Co 1997. COLES B H.vet. In: RUPLEY AE (ed): Manual of Avian Practice. Philadelphia: WB. 6. Potential Pathogens Recovered from Upper Respiratory Tract of Psittacine Birds. DORRESTEIN GM and QUESENBERRY K (eds): Avian Medicine and Surgery. Saunders Company 1997. in a contrary. especially because of prior good health status of the bird and no contact what so ever with any other birds species or animals. and rise a question why did infection with severe clinical signs appeared. BRENNER DJ and FARMER III JJ.uga. Heinzelova 55. CLUBB SL. BALOWS A. Bacteriology. Common Diseases and Treatments. Horvatek. Croatia Email: horvatek@vef. Therefore our finding is not unusual. GERLACH H.280. 4 CITATION INDEX 1. Philadelphia: WB. 3. DVM Faculty of Veterinary Medicine.edu) KELLY MT. but according to JESUS (1998) Klebsiella species are isolated from healthy birds and from birds with upper respiratory tract infection. HAUSLER WJ AND SHADOMY HJ (eds): Manual of Clinical Microbiology Fourth Edition.hr 500 . In: LENNETTE EH. Bacteria. 4. symptoms worsened. In: RITCHIE BW. Munich 2001. This case is presented here because in a last few years K. without any contact with other birds. Past Klebsiella oxytoca contamination of a drinking water supply. AUTHORS ADRESS D. HARRISON GJ and HARRISON LR (eds): Avian Medicine: Principles and Application.983. 263 . DORRESTEIN GM. JESUS O S and DUARTE CORREIA J H. Lake Worth: Wingers Publishing 1994. 264 . Enterobacteriaceae. oxytoca wasn’t found in patients in our Clinic. 2. RUPLEY AE. resulting in an increased juvenile mortality in a variety of avian species in a zoological collection. In: ALTMAN RB. Our recommended therapy with enrofloxacin results in complete recovery. 6. 5. It was housed in a large stainless-steel cage. Washington: DC: American Society for Microbiology 1985. Proc Euro Assoc Avian Vet. University of Zagreb. 949 . 255 .277. tracheobronchial tree. fungal and bacterial cultures. Polychondritis has been described in humans and feline patients and can involve the ears. the fibrocartilage at axial sites and the cartilage in the tracheobronchial tree. multisystem disorder of characterised by recurrent episodes of inflammation of the cartilaginous tissue. 1 INTRODUCTION Polychondritis. also know as relapsing polychondritis (RP) is an uncommon. United States of America CHRONIC POLYCHONDRITIS IN AN AFRICAN GREY PARROT (Psittacus erithacus) T. ribs and pelvis. K. nasal and laryngotracheal cartilage and 501 .Computed tomography . Immune mechanisms are presumed to be of aetiopathogenic importance. Classic features are repeated inflammatory episodes involving auricular. All types of cartilage may be involved.Angell Animal Medical Center. Relapsing polychondritis can also inflame other proteoglycan-rich structures. Systemic symptoms including fever. The condition can involve hyaline cartilage. such as the eye. lethargy and weight loss are common. At presentation the bird lacked the external nares and the nasal structures were deformed with severe loss of the normal cartilaginous tissue. including the elastic cartilage of the ears and nose. elastic cartilage or fibrocartilage. The disease condition of chronic atrophic polychondritis was obtained from diagnostics including blood work. nose. larynx. Polychondritis is a disease of unknown aetiology characterised by inflammation and degeneration of the cartilaginous structures. CT scan and biopsy of the nasal cartilage for histopathology. the hyaline cartilage of the peripheral joints.Cartilage .Psittacine ABSTRACT A twenty year old male Congo African grey parrot presented for a chronic history of progressive deformity of the external nares and nasal vestibule. Dipl ABVP-Avian Practice KEYWORDS Polychondritis . Vasculitis affecting skin or internal organs may occur. DVM. heart. This is the first known case report of polychondritis in a psittacines bird. Polychondritis is presumed to be an autoimmune disease. Boston. blood vessels and inner ear. Ritzman. This criteria state that the patient must have 3 of the following 6 clinical features for diagnosis. Contiguous axial 1 mm images were obtained from the rostral premaxilla through the level of the orbit. fibrosis and sequestration consistent with polychondritis. d) ocular inflammation. e) respiratory tract chondritis. and includes the use of NSAID’s. coli and Citrobacter freundii which was due to secondary bacterial infection of the exposed tissue. Most human patients develop some degree of disability such as bilateral deafness. Histopathological findings with polychondritis demonstrate chondrolysis. Computed tomography (CT) of the head was performed. (1976) and has been modified several times. f) and/or audiovestibular damage. There is no age. Polychondritis has been described in felines (GERBER et al. IX. The cartilage matrix is eventually destroyed and replaced by fibrous connective tissue. and XI are present in 25-70% of human patients with RP. joints or ocular tissues. Bloodwork testing was performed and the results are included in the poster. b) non-erosive seronegative inflammatory polyarthritis. The clinical features include: a) bilateral auricular chondritis. maxillary and frontal bones. 2002) and the condition in cats is usually characterised by a folding deformity elastic cartilage of the pinnas (GERBER et al. and cardio-respiratory problems. phonation difficulties. Skull radiographs performed under general anesthesia showed cartilage and bone defect centered at the nasofrontal hinge. Biopsy of the affected cartilaginous tissues with histopathology may aid in the diagnosis. Various studies indicate that circulating antibodies to cartilage-specific collagens types II. A large lesion was visibly centered at the nasofrontal hinge with loss of portions of the premaxillary.occasionally. fungal culture and fluid cytology. and perichondritis. 2 CASE REPORT A twenty year old wild caught male Congo African grey parrot (Psittacus erithacus) presented for a chronic history of deformity of the external nares and nasal vestibule. Multiple biopsies were collected from the exposed hyaline cartilage of the naris-nasal vestibule (left and right) and submitted for histopathology. Medical therapy is based on the understanding of other connective tissue or autoimmune diseases. c) nasal chondritis. Diagnosis in humans is made based on diagnostic criteria first proposed by MCADAM et al. 502 . Potentially serious complications include laryngotracheal collapse and or stenosis. The inner nasal vestibule was visibly occluded with dried mucopurulent discharge. gender or ethnic pattern to this disease in humans. At presentation the bird lacked external nares and the nasal vestibule was deformed with severe loss of the normal cartilaginous tissue. impaired vision. prednisone and/ or immunosuppressive medications to help control the symptoms. and their enclosed nasal passages. The bacterial culture produced abundant growth of E. There is no known cure for polychondritis Treatment is directed at reducing the inflammation and progressive destruction of the affected tissues. valvular heart disease and large-vessel aneurysms. A nasal flush was performed and this fluid sample was collected for aerobic bacterial culture and sensitivity testing. 2002). This is the first known case report of polychondritis in a psittacine bird. Fungal cultures were negative. Histopathology showed multifocal degeneration and necrosis of the hyaline cartilage with fracture. chondritis. 215. Antibiotic therapy is provided as indicated for the recurring secondary bacterial infections based on culture and sensitivity results. TSCHARNER C VON et al. MCADAM LP. Relapsing polychondritis: prospective study of 23 patients and a review of the literature. Dipl ABVP-Avian Practice Angell Animal Medical Center. Boston.This patient is being treated currently with non-steroidal anti-inflammatory therapy (meloxicam 0.194. 3 CITATION INDEX 1. MA 02130.org 503 . Medicine (Baltimore) 1976. AUTHORS ADDRESS: Tracey K. 55(3): 193 . There has been no appreciable progression of the disease in this patient over the past 20 months. DVM. such as systemic mycosis in this susceptible species. 2. United States Email: tritzman@angell. CROTTAZ M. Ritzman. J Feline Med Surg 2002.5 mg/kg PO SID) as well as regular nasal flushing and debridement to keep the exposed nasal tissues clean. Boston 350 South Huntington Ave. GERBER B. Immunosuppressive therapy was considered but not instituted in this patient due to the concern for increasing risk of secondary infection. Feline relapsing polychondritis: two cases and a review of the literature. BLUESTONE R and PEARSON CM. O’HANLAN MA. 4(4): 189 . MS. 1 INTRODUCTION An understanding of the tolerance of normal tissues included in the treatment field is essential in radiation treatment planning and the initial decision as to whether radiation should be used for cancer treatment. PhD. usually poor. In fact. and Kenneth Latimer. 60. United States of America RADIATION TOLERANCE DOSES OF CUTANEOUS AND MUCOSAL TISSUES IN PSITTACINE BIRDS USING COBALT-60 TELETHERAPY G. DACZM. ACVR. Stephen HernandezDivers BVetMed. DVM Dipl ABVP-avian. DVM. success (MANUCY et al. Ingluvial mucosae and the skin over the ingluvies of nine Psittacine birds were irradiated in 4 Gy fractions to a total dose of either 48. or 72 Gy using an isocentric Cobalt-60 teletherapy unit. 504 . DVM. Attempts in the past to use external beam radiotherapy (RT) for cutaneous neoplasms in birds have met with variable.College of Veterinary Medicine University of Georgia. neoplasms are more commonly reported in the budgerigar than in any other domestic animal (BAUCK 1992). Heather Wilson. Neoplasia is a common problem in pet birds. DZoo Med. MRCVS. DVM. Dipl. Royce Roberts. Acute and chronic radiation effects were measured. Diplomate ACVIM (oncology). Neither doserelated acute nor chronic radiation effects could be detected grossly in cutaneous or mucosal tissue over a 9 month period. Histopathology demonstrated only minimal acute changes in the high dose group. treatment for avian patients with neoplasia has lagged far behind the technology available for other companion animals. Northrup. Despite this. DACVP KEYWORDS Cobalt – Radiation – Tolerance – Ingluvies – Skin – Ring-necked Parakeets Psittacula krameri ABSTRACT The purpose of this study was to demonstrate what level of Cobalt-60 radiation could be tolerated by the cutaneous and mucosal tissues of normal ring-necked parakeets (Psittacula krameri) in order to provide more effective treatment for tumours that have been shown to be radiation responsive in other species. Nicole C. WILSON 2000). This study indicates that doses of 72 Gy given in 4 Gy fractions may be used safely. In these birds.1998. the histology was similar to that of the control group. all birds received a score of 0 (no change compared to baseline). 3 RESULTS Grossly. 2004) However. The other 3 groups were irradiated to a total dose of 48. all 3 birds in the high dose group (72 Gy) had minimal radiation-induced epidermal changes. but radiation therapy was not performed. influence of fractionation. The goal of this study was to determine the tolerance doses of cutaneous and mucosal tissues of healthy psittacine birds in order to provide more effective treatment for neoplasms that have been shown to be radiation responsive in other species. (ZALLINGER and TEMPLE 1998) Dose response. The field size was 2 x 2 cm for all subjects. (GILLETTE et al. or in the case of the control group at week 6. However. These lesions were clinically insignificant. lateral treatment portals were used for each treatment. each composed of 3 adult. 4 DISCUSSION The principle of radiation therapy is to maximize the radiation dose to the tumour and tumour-harbouring tissues while minimizing radiation exposure to surrounding healthy tissues. with no clinically significant dose-related acute or chronic radiation effects on 505 . 1995) Biopsies of skin and ingluvial mucosa (3-4 mm) for histologic analysis were obtained at the end of each group’s treatment period. on the VRTOG radiation scoring scheme at the end of radiation treatment and throughout the duration of the 9 month study. and 72 Gy respectively using an isocentric Cobalt-60 teletherapy unit. parallel-opposed. clinically healthy birds. The parakeets were monitored for a total of 9 months for chronic effects. having crop and skin biopsy. there have been no controlled studies of external beam radiation therapy in avian species. A total of 12. One study demonstrated that of all domestic species mortality and morbidity due to acute radiation syndromes was lowest in chickens. Body weights remained stable throughout the study. Equally weighted. Wednesday. as did appetite and attitude. Lesions were graded according to the Veterinary Radiation Therapy Oncology Group acute and chronic radiation morbidity scoring criteria. There was no histologic evidence of radiationinduced tissue alterations in either the low (48 Gy) or the medium dose group (60 Gy). 2 MATERIAL AND METHODS Twelve male and female Indian ring-necked parakeets ranging from one to two years of age were divided into 4 randomized groups. centered over the ingluvies. One group served as controls. and Friday until the total prescribed dose was achieved over a 4 to 6 week period. 60. and normal tissue injury has been evaluated extensively in other companion species over the past 25 years. 15. Freeman 1999. or 18 fractions of 4 Gy each were given on a Monday. (ASTASHEVA et al. MANUCY TK. United States Email:
[email protected]. 1992. J Assoc Avian Vet 1998.normal cutaneous or mucosal tissue in Indian ring-necked parakeets. Radiation therapy for hemangiosarcoma in a budgerigar. 12(3):158 . growth. Integumentary neoplasms in psittacine birds: treatment strategies. DVM. Seminars in Avian and Exotic Pet Medicine. The physiologic response of domestic animals to ionizing radiation: a review. University of Georgia. ABVP (Avian) Department of Small Animal Medicine. et al. Vet Rad & Ultrasound 1998.213. 1(2): 65 . Sem Vet Med and Surg 1995. FREEMAN KP. et al. 6. This project was approved by the University of Georgia’s Animal Care and Use Committee. Normal tissue tolerance and management of radiation injury. GRAHAM J. 2. 211 . 7. GILLETTE EL. ASTASHEVA NP. 13(1): 40 .503. KHRAMTSOVA LK and FINOV VP. Portland 2000. Lower doses of radiation may be insufficient to provide effective tumour control. ADMAS WH. GA 31522. Athens. BENNETT A. 5 CITATION INDEX 1.46. LARUE SM and GILLETTE SM.214.edu 506 . Heather Wilson. 3.uga. Radiata Bio Radioecol 2004. ROBERTS RE. Dipl. and productivity of the broiler chicken. 4. 39 (6): 495 . Proc Assoc Avian Vet. J Assoc Avian Vet 1999. AUTHORS ADRESS G. Effects of gamma-irradiation on survival. ZALLINGER CV and TEMPLE K. et al. WILSON GH. 10(3): 209 . A clinical approach to neoplastic disease in the pet bird. GREENACRE CB. Squamous cell carcinoma of the mandibular beak in a Buffon’s macaw (Ara ambigua). BAUCK LA. ANNENKO BN.72. College of Veterinary Medicine. HAHN KA. 5. 44(1): 43 . All data collected are analyzed and recorded in a database. the French Game and Wildlife Agency) and the CNERPAS (that became AFSSA – LERRPAS). 5 500 birds were necropsied by SAGIR. GAILLET. intoxications (bromadiolone. fowl pox. drowning. It gives informations on the sanitary background of wildlife and on the distribution of pathogens that are common to humans and domestic animals. … 1 INTRODUCTION Created in 1986 by the ONC (that became ONCFS) and the CNERPAS (that became AFSSA). listeriosis. their geographical origin.R. FRANCE ONCFS. DVM KEYWORDS SAGIR . J. One of the advantages of SAGIR is its national coverage. The presentation first shows the different species analyzed.LERRPAS.France ABSTRACT SAGIR is a national surveillance system of wildlife diseases. This network groups terrestrial mammals (about 5/6 of the analysis) and birds (about 1/6). BARRAT.created in 1986 by the ONC (that became ONCFS.E. The drawback is the sampling method: the necropsy of animals is carried out on a voluntary basis according to the interest of the hunters in each case. The main causes of death are traumas. electrocution. aspergillosis). bacterial diseases (botulism. chloralose and various inhibitors of cholinesterase). Since 1989. TERRIER. J. the yearly changes in the analyses. Nowadays. A yearly report is done and published in the BIPAS (“Bulletin d’Information sur la Pathologie des Animaux Sauvages”). salmonellosis) and parasitic diseases (trichomonasis. DVM. SAGIR is a national surveillance system for wildlife diseases.EAAV 2005 AFSSA . DVM. FRANCE DISEASES FOUND IN FRENCH BIRDS ACCORDING TO SAGIR DATABASE M. nearly 500 birds are recorded yearly.network – diseases – birds . 507 . Some 3 000 to 4 000 animals are necropsied each year. chlamydiosis. Some pathological findings are selected: avian tuberculosis. the gamekeepers (ONCFS). the Ministry of Agriculture.intoxications: bromadiolone. 2. chlamydiosis. pseudotuberculosis. fowl pox.bacterial diseases: botulism. Columbiformes.LERRPAS BP20 54 220 MALZEVILLE / France e-mail: me. . diurnal birds of prey. More than half of birds are game birds. but about 1/3 of them are protected species. This may be due to the small number of necropsies of birds done yearly by some laboratories. AUTHORS ADDRESS: M. . AFSSA-LERRPAS. more than 5600 birds were necropsied by SAGIR. even if it is with different levels of implication. Most of the results have been obtained between 1993 and now. aspergillosis. and not necessarily correlated with the identified disease. the toxicology laboratory of the National Veterinary School in Lyon (ENVL) and other specialized laboratories. the cause of death stay unexplained.parasitic diseases: trichomonasis. the National Parks. DVM AFSSA . 3. listeriosis. the Local (“Départemental”) Veterinary Laboratories (LVD) and the Local (“Départemental”) Federations of Hunters (FDC). Interesting diseases. chloralose and various inhibitors of cholinesterase. Many lesions are observed. Causes of death of the birds. drowning. Their distribution is as follow: web-footed birds.terrier@nancy. Some interesting pathological findings are selected: avian tuberculosis. One of the advantages of SAGIR is its national coverage. Most of the registered results come from SAGIR network.E.2 MATERIAL AND METHODS SAGIR is organized as a network including ONCFS. salmonellosis mostly due to Salmonella arizonae . scientific teams (for example CNERA of the ONCFS). Since 1989. Charadriiformes. Terrier.traumas: predation and road accidents. The annual number of birds analyzed now is about 500. other various species. Anseriformes. The drawback is the sampling method: the necropsy of animals is carried out on a voluntary basis according to the interest of the hunters in each case. … 4 DISCUSSION In about ¼ cases. which means that the whole country participates in the network. Galliformes. .afssa. but we also get results from the FDC without a SAGIR form. … The birds come from 93 different “départements”. another linked one is the reactivity of the network. The main identified causes of death are . the foresters (ONF). 3 RESULTS 3. 1. electrocution. cranes. The SAGIR network needs specialized avian veterinarians to refer difficult cases to. passerines.fr 508 . coccidiosis. The FDC pays for the different routine analyses. Conical: diurnal birds such as Falconiformes. 1 INTRODUCTION The eye is the most important sensory organ in birds. Common ophthalmological examination procedures will be discussed together with the most frequent ocular disorders. the eye may be seen as an important diagnostic criterion in avian medicine. 1. France CLINICAL APPROACH TO OPHTHALMOLOGIC DISORDERS IN BIRDS F. Ultraviolet (UV) light perception probably plays an important role in communication based on plumage UV reflection. 509 . DVM KEYWORDS Birds – Ophthalmology – Eye ABSTRACT Anatomical and physiological characteristics of the bird eye are described.Clinique vétérinaire de l’Arche. Rival. Thus. Visual fields are up to 360°. ocular lesions are an expression of systemic diseases. Visual acuity is 2-8 times higher compared to mammals. 2 PHYSIOLOGICAL AND ANATOMICAL CHARACTERISTICS The avian eye show a high specialisation as an adaptation to fly. In birds. Shape : Three basic shapes are typical: Flat : diurnal birds such as Columbiformes. Tubular : nocturnal birds such as Strigiformes. Valence. Maintained by 10 to 18 scleral ossicles. Eye examination should be an integral part of the general clinical examination protocol. more often than in mammals. replaced by the pyramidalis and quadratus muscles innervated by cranial nerve VI.Nasal salt gland in some species Complete decussation of chiasma opticus (no consensual papillary reflex) Anterior segment: Striated intraocular musculature (no sensitivity to SM or PSL. Evaluate the palpebral reflex. thin.1. Avascular retina. . No retinal tapetum lucidum. Pecten oculi (vascular prominence of variable shape). 510 . Adnexal structures: eyelids: . 7. Iridocorneal angle well developed Lens with annular pad at ist equator (anulus pulvinus) Posterior segment: Vitreous body: large and transparent.Small and absent glandulae lacrimaliae . Rods and cones (inclusive special UV cones).No Meibomian glands. 4. 3. - 5. - 3 CLINICAL EXAMINATION A complete ocular examination is composed of three parts : 1. transparent and mobile. 3.Nictitans membrane. Retractor bulbi muscle complex absent. adnexa. 6.3. - Ocular motility limited. anterior and posterior segments are examined. . the direct pupillary light response. 2.Two lacrimal puncta and a nasolacrimal duct .2. . Morphologic ocular examination : external eye.2 Functional examination: it assesses vision directed behavior.Large Harderian gland (glandula lacrimalis membranae nictitantis) . lacrimal glands: .Lower eyelid with a fibrous plate (tarsus). no reliable pupillary reflex corresponding to light stimuli).Filoplumes (mammalian cilia). History. Mydriasis can be achieved by general anaesthesia with APA (air sac perfusion anaesthesia).3 Classification by localisation. 511 . while G.4. 563 . DORRESTEIN GM and QUESENBERR YK (eds): Avian medicine and surgery. New Orleans 1999: 335 . KERN TJ. However it has to be administered directly into the anterior chamber by paracentesis to be more effective. KORBEL RT.589. In: ALTMAN RB. Proc Assoc Avian Vet. Philadelphia: WB Saunders 1997. Mydriatics have little or no effect in the avian eye due to the striated musculature. Neuromuscular blocking agents such Tubocurarine 3% may be used. Ophtalmoscopy : induction of mydriasis is indispensable for the examination of the posterior eye segment. This anaesthetic technique is excellent for ophthalmologic examination and head surgery. Rival. OTHER EXAMINATIONS Tonometry : standard reference values : 9-22 mm Hg (corneal diameter > 9mm). AUTHORS ADRESS F. Classification by aetiology. 2. DVM. Avian ophthalmology – a clinically-oriented approach. 5 CITATION INDEX 1. The combination médétomidine/ketamine (275 microg/kg. at 10mg/kg) IM is also a good anaesthetic choice for head surgery. Physiological bacterial flora contains G+. Schirmer tear test or Tevetest (in study): difficult because of the interspecific variations.1. - 4 EYE DISORDERS IN BIRDS (see poster) 4.350. 2. Disorders of the special senses: The eye.are indication for pathological conditions. Président du GENAC. Cytology and bacteriology : corneal or conjunctival cytologic specimens may be examined. CLUBB SL. 2.2 Classification by clinical signs. CES d’Ophtalmologie Clinique vétérinaire de l’Arche. It is usually caused by local ischemic necrosis created by the inflated cuff of the endotracheal tube. This paper reviews cases of post-intubation stenosis.intubation stenosis (PIS) is the commonest reason for tracheal resection and anastomosis in man (WAIN 1975).Avian and Exotic Animal Clinic of Indianapolis. M. 1975). inflammation and necrosis of tracheal cartilage (KLAINER et al.Post-intubation . Cheshire. Stanford BVSc MRCVS KEYWORDS Tracheal stenosis . the possible aetiologies of the condition. rough intubation and 512 . pre-existing inflammation in the trachea. treatment and prevention protocols. The use of chemical disinfectants to disinfect tubes between patients has been suggested as an alternative aetiology for stenosis. medical and stent placement may be successful. reduction of cuff inflation pressure and the use of sterile single use endotracheal tubes. Stenotic tracheal lesions in birds secondary to tracheal intubation typically appear as a severe inflammatory response in the superficial tissues rather than cartilage necrosis. In mild cases mechanical dilation. 1 AETIOLOGY OF POST-INTUBATION STENOSIS IN PSITTACINE BIRDS Post. The condition can develop after very short intubation procedures. The condition is prevented in humans by careful tube selection (both length and diameter). United Kingdom POST-INTUBATION STENOSIS IN PSITTACINE BIRDS A. If the trachea cartilage is damaged then surgical intervention is required with a reported success rate of 93. AVBP-Avian. laser. This may be a reflection that cuffed tubes are contraindicated in avian medicine due to the anatomy of the avian trachea with complete cartilage rings. Other possible aetiologies would include inappropriate tube size. The condition is also reported in psittacines with an apparent predilection for macaw and cockatoo species.Psittacine ABSTRACT Tracheal stenosis is a commonly reported sequel to endotracheal intubation in humans. Lennox DVM Dipl. Indianapolis. D.7%. M. United States of America Birch Heath Veterinary Clinic. Histopathological examination of affected tracheas suggests an acute inflammatory reaction in birds compared with man where cartilage necrosis is invariably reported. Histopathological lesions include degeneration. Always carefully select the tube diameter based on where the tube tip will rest in the trachea. 513 . This might also reflect the difficulty in diagnosing trachea stenosis in the smaller psittacine birds leading to under reporting of the condition. Management of tracheal trauma in birds. Two rings on either side of the tracheotomy site should be included in each suture.713. 3: 693 . Chest Surg Clin N Am 2003. Proc Australian Assoc Avian Med 2001. Intubate carefully under the appropriate plane of anaesthesia. It is useful to note that the condition has not been reported after tracheal endoscopy procedures suggesting the condition is due to prolonged contact between the tube and tracheal mucosa. et al. WAIN JC. 1975. Proc Assoc Avian Vet. LENNOX AM. Permanent tracheotomy has been described in psittacine birds and may be considered in unsuccessful resection cases. BENNETT RA. placing knots externally to the tracheal lumen. 2 MANAGEMENT OF POST-INTUBATION STENOSIS IN PSITTACINE BIRDS Treatment of tracheal stenosis can range from conservative therapy. Of the nine cases reviewed 5 died or were euthanased within 6 months of repair. 4. Use sterile or adequately rinsed tubes. Avoid long intubation or anaesthetic times. New Orleans 2004. Incisions should be made between tracheal rings and closed with 4-0 to 8-0 monofilament absorbable suture. minimal number of sutures with adequate control of secondary infection and inflammation (PYE 2000). The diameter of the normal trachea in both species reduces by up to 55% along its length (LENNOX 2004). 3 CITATION INDEX 1. Am J Med. Stenosis at the point of anastomosis is reported as a complication of surgery. resection of the stenosis followed by anastomosis is normally required (CLIPPINGER and BENNETT 1998). PYE GW. Avoid bending the tube or trachea during the procedure. Post-intubation tracheal stenosis. Tracheotomy. 3.mechanical irritation due to the end of the tube touching the tracheal wall. The use of cuffed tubes should be avoided in birds.343. Surface alterations due to endotracheal intubation. Of the nine cases reviewed for this paper 5 were macaw species and three cockatoos. The selection of an endotracheal tube based on glottis diameter might therefore be too large in these birds. Ventilate gently using the minimal pressure to achieve adequate ventilation. 58(5): 674 . In the opinion of the authors PIS could be avoided by following these simple principals. 13(2): 231 . Surgery of the avian respiratory system. Avian respiratory tract surgery. Adequate control of inflammation is a prerequisite for successful management of tracheal trauma. 2. TURNDORF H. including antibiotics and anti-inflammatory medications. to complete tracheal resection and anastomosis depending on the severity of the condition (BENNETT 2001). 339 . WU WH.683. 5. Vet Clinics North Am 2003. Successful surgery depends on atraumatic tissue handling. 103-108.246. KLAINER AS. careful placement of sutures. CLIPPINGER TL and BENNETT RA. Lennox Avian and Exotic Animal Clinic of Indianapolis. J Av Med Surg 1998.com 514 . United States of America Email: BirdDr@aol. Successful treatment of a traumatic tracheal stenosis in a goose by surgical resection and anastomosis. 12: 243 . Indianapolis.6.247. AUTHOR ADDRESS M. Monks BVSc (Hons) MACVSc (Avian Health) CertZooMed MRCVS N. The most frequently identified tick species was Ixodes frontalis. Shernacre Cottage. This report aims to discuss the species of ticks infesting birds in the UK. Babesia and Borrelia. Forty-two birds (60%) had adult female I.Great Western Referrals. Swindon.Haemorrhage . infection. Malvern. Forbes BVetMed CBiol MIBiol DipECAMS FRCVS M. spleen and aspirate from tick attachment site using a silica cartridge-based kit (MacheryNagel) and tested at the AcarusTM laboratory (Langford. to test (via PCR) for tick-borne infections. requesting samples from wild and captive birds. PCR tests were uniformly negative for Ehrlichia. It aimed to characterise ticks infesting birds in the UK. test tick-borne infections via PCR. Bartonella. parasitic. Ticks were identified according to standard keys.Tick . ocular. 2 MATERIALS AND METHODS Submission kits and questionnaires were sent throughout the UK. United Kingdom IXODES FRONTALIS AS A CAUSE OF AVIAN DISEASE IN THE UNITED KINGDOM D. UK). unknown). of which 25 birds (59.Oedema ABSTRACT This study obtained samples from 70 birds with attached ticks or signs consistent with tick related syndromes. Fisher BVetMed CBiol MIBiol DipEVPC MRCVS KEYWORDS Ixodes frontalis . PCR was performed on ticks. miscellaneous. 1 INTRODUCTION Avian tick-related syndrome (TRS) is a syndrome in which birds with an attached tick present acutely lethargic and depressed or dead with mild to severe subcutaneous haemorrhage and oedema on the head or neck.5%) had signs referable to tick related syndromes. and to examine the correlation between tick infestation and disease. frontalis ticks present. Clinical presentations were categorised as associated with TRS or not (trauma. and to determine the correlation between tick infestation and disease.Avian . 515 . blood. 1996). frontalis females and clinical syndrome or outcome. PCR testing was subsidized by the AcarusTM laboratory and their staff. Our study would seem to indicate that there is a strong correlation between attachment of adult female I. may be responsible for TRS. The host response to tick infestation may contribute to the aetiology of TRS. frontalis had previously detached from the birds. Bartonella. Avian tick-related syndrome is clearly an area requiring further research. engorged adult I. However. 1991). All samples submitted to PCR tested negative for DNA of Borrelia burgdorferi sensu lato. Adult female I. Bartonella species and Ehrlichia species with the AcarusTM battery of tests. In total. Although it is possible that fullyengorged adult I. in this study one case of swelling was associated with three I. while no submissions were received in January or February. especially Drs Martin Kenny and Susan Shaw. 516 . 5 ACKNOWLEDGEMENTS The project was supported by grant funding from BSAVA Petsavers. Signs of TRS were also found with the presence of three I. On PCR no sample tested positive for B. are thanked for their work on this project. All of the adult and nymph ticks were female. In total. the absence of an adult tick on these cases raises the possibility that other life cycle stages may be capable of causing disease in certain hosts. 4 DISCUSSION Ixodes frontalis is a specific avian parasite and has been documented to cause TRS in birds (CHASTEL et al. of which 25 (59. Tissue oedema and haemorrhage associated with I.5%) had signs of TRS. frontalis nymphs and an unspeciated larva. including viruses. Sixty-eight of those were from birds with attached ticks. Additionally.3 RESULTS Submissions were solicited from April 2000 to January 2004. It is possible that other pathogens. frontalis attachment has only been previously reported with multiple engorged adult female ticks (CHASTEL et al. The majority of submissions occurred in August and September. Babesia species. 1991). There was no correlation between numbers of attached. 161 ticks were examined. some ixodid species produce specific salivary toxins (LUTTRELL et al. frontalis and TRS. 70 samples were tested. frontalis were present on 42 birds. Most birds presented with one or two ticks. Ehrlichia and Babesia in our study.burgdorferi sensu lato. frontalis nymphs while another was associated with an unspeciated larva. monks@gwreferrals. 32(1): 133 . LUTTRELL MP. AND MERTINS JW. County Park Business Park. J Wildl Dis 1996. Email: d.32. United Kingdom.uk 517 . Avian tick paralysis caused by Ixodes brunneus in the southeastern United States. CHASTEL O and BEAUCOURNU JC. Swindon. Pathological effects. Annales de Parasitologie Humaine et Comparee 1991. 66(1): 27 .co. SN1 2NR. CHASTEL C. vector role and new host of Ixodes Pari (= Ixodes frontalis) Acari Ixodoidea Ixodidae. Shrivenham Road. AUTHORS ADDRESS Deborah Monks Great Western Referrals. CREEKMORE LH.136. 2. GUIGUEN C.6 CITATION INDEX 1. ABSTRACT Research on three different Cuban Amazon (Amazona l.Department of Animal Production. S. WRIGHT 2001). in particular of nestlings. Gálvez Aguilera5 KEYWORDS Amazona l. 12 of Dermanissus spp.Wild populations Haematology – Parasitology . monitoring health conditions and supporting breeders. and very few endo and ectoparasites were found (5 cases of Ascaridia spp. These parrots are traditionally considered companion birds in the houses of Cubans. Bert1. University of Turin. leucocephala) populations within a conservation project in Cuba. X. presence of parasites and prevalence of Chlamydial antibodies were evaluated. THE ROLE OF AVIAN MEDICINE IN A PARROT CONSERVATION PROJECT ON AMAZONA LEUCOCEPHALA IN CUBA : PRELIMINARY RESULTS E. Cruz López1.). Farming these birds is an interesting alternative to the capture of wild parrots. I. 1 INTRODUCTION Amazona l. which are sold in Cuba (WILEY et al. of Animal Health. birds raised in captivity could satisfy the demand for domestic birds without affecting the size of wild populations. Tomassone1. G. Dep. poaching still occurs. is described. Despite the international laws that forbid the capture and trade of these birds. 518 . C. Italy. Veterinary medicine can have an important role in species preservation. Alvarez4 . J.Chlamydial antibodies. Gonzáles Navarrete3.Breeding . leucocephala . Forneris1. Acosta Guevara1. included in the IUCN Red List of the endangered species (GRAJAL 2000).Conservation . E. Soto Piñeiro2. Epidemiology and Ecology1. leucocephala is a Cuban endemic subspecies. we investigated the health status of A. M. No Chlamydial antibodies were detected. National Ornithological Association of Cuba2. Agricultural University of La Havana – Cuba3.. leucocephala parrots in Cuba from three distinct populations. Within a conservation project. Zapata Swamp National Park4 National Organization of Flora and Fauna Conservation5 Cuba. Haematological parameters. 1995. L. A control plan for parasites is advisable in breeding centres. Few endo and ecto-parasites were found: including no cases of haemoparasites. No differences in haematological parameters were noted between the second and third groups (Zapata Swamp National Park and La Havana captive birds respectively) or between juveniles and adults. Israel). Haematocrit and red blood cells count were carried out in a field laboratory. at the limits of the tropical forest and were in contact with wild birds.2 MATERIALS AND METHODS From March to September 2004. both to study the physiological characteristics of the species and to recognize the situations when intervention may be necessary. Group 2 comprised 50 birds (18 juveniles and 32 adults) living in a breeding station in the Zapata Swamp National Park (ZSNP). probably due to the location of the aviaries. faecal and ectoparasite samples were collected.. We believe that the role of veterinarians is important in a parrot conservation project. in nests with intense infection. On the contrary. leucocephala in Cuba. 519 . Chlamydial antibodies were investigated with the ImmunoComb® Avian Chlamydophila psittaci Antibody test kit. Our study is the first of the kind carried out so far on A. Average haematological values corresponded to published values for other Amazon species. 5 cases of Ascaridia spp. in which only nestlings have been sampled. moreover. Blood. In the case of the LIER population. The presence of ectoparasites was noted in the ZSNP. and 12 cases of Dermanissus spp. In the wild population. Another important aspect is providing suggestions to owners during the clinical visits. the aviaries were located in an open space. in order to increase the life expectancies of subjects kept in captivity. In this centre. 4 DISCUSSION Hematological parameters of adult and juvenile Cuba Amazons kept in captivity correspond to those reported for other Amazon species (FUDGE 2000). three parrot populations were studied. 3 RESULTS No Chlamydial antibodies were detected. According to our data. chick condition was poor and an intense anemia was noted. endoparasitic infections do not appear to be very frequent in this species and cause little damage. The nestlings of the wild population in “Los Indios” showed lower blood parameters (Table 1). ectoparasites seem to have a significant impact. results are difficult to interpret. thanks to a donation by the manufacturers (Biogal Galed labs. they cannot be considered definitive and a reference range could not be established because of the small sample size. Group 3 comprised 55 Cuban Amazons (21 juveniles and 34 adults) kept in captivity in private houses in La Havana. Group 1 comprised 15 nestlings from a wild population living in the “Los Indios” Ecological Reserve (LIER) of Juventud Island. 5 LITERATURE CITED 1. Status survey and conservation Plan 2000-2004. Cons Biol 2001. et al.bert@unito. Switzerland and Cambridge. GÀLVEZ X. Nest site ecology of the Cuban Parrot. In: SNYDER NMC et al. Via Leonardo da Vinci 44. Nest poaching in neotropical parrots. AUTHORS ADDRESS Dr. GNAM RS. 3. UK. Italy Email: Elena. Laboratory Medicine . 2. Report of activities during 1995. Philadelphia. IUNC.55 WILEY JW. GRAJAL A. Neotropics.Avian and Exotic Pets.it 520 . WPTI Grant 94/95. 2000: 2 . University of Turin. DVM. PhD Department of Animal Production. (eds): Parrots Action Plan. Elena Bert. N. 2000: 486. 10095 Grugliasco (Torino). 4. USA. Sound Company. Gland. FUDGE AM.720. Epidemiology and Ecology. WRIGHT T.010: 17. 3: 710 . 18 160-175 5000-17000 125-250 6020-4408 176 5312 48.62 0-1 0-2 Eos (%) 0 0 0 Htc (%) Nestlings 29. leucocephala (n=15) (Reference values from FUDGE 2000).71 35-72 31-71 42 26-63 20-67 MCV (fL) WBC /µL Heter (%) Linf (%) Mon (%) 1.10 2. RBC (x103/ µL) 1.26 – 2.69 1-2 0-2 Bas (%) 0.29 521 Range 25.45-3.Table 1. Haematological parameters of the wild population of A.5 Reference 41-53 .34.5 .70 1. . J. Handling and Restraint of Parrots M. European Association of Avian Veterinarians Conference The texts will be given to the participants during the practical labs L.P. Dorrestein. J. Hatt. Avian ophthalmology: principles and applications G. Morrissey. Raptor biomedicine 523 . Forbes.T. Hooimeijer. Basic and advanced avian cytology and haematology M. basic and advanced R. Avian soft tissue surgery S. N. Behavior. Lithtenberger. Avian endoscopy N. Harcourt-Brown. Orosz. Korbel. J. Crosta.A.M. H.Practical laboratories. Speer. Péricard. J. Bohnet.M. Lierz. Emergency and critical care in birds J. André Avian clinical imaging.M. Pendl. B. Orthopaedic surgery in birds N.