ISSN: 2084-3577TMKARPIŃSKI PUBLISHER Journal of Biology and Earth Sciences ORIGINAL ARTICLE BIOLOGY Influence of synthetic and plant extracts on biochemical changes in the gonadal tissue of snail Lymnaea acuminata Vatsala Singh, J.N. Tiwari Department of Zoology, TD College Jaunpur, UP, India ABSTRACT Fasciolosis is now recognized as an emerging human disease. Control of intermediate host is an effective tool for control of liver fluke infection. In present study effect of sublethal treatment (20% and 60% of 24h LC50) with permethrin, cypermethrin, Allium sativum and Abrus precatorius plant extract caused a significant (p<0.05) reduction in the protein, amino acid, DNA and RNA level in the gonadal tissue of snail Lymnaea acuminata. Maximum reduction in the level of protein (33.55% of control), amino acid (1 6.21 % of control), DNA and RNA (32.96 % and 1 3.70% of control) with the treatment of 60% of 24h LC50 of permethrin after 96h exposure periods. Among plant extract Abrus precatorius is more pronounced than Allium sativum. Key words: Fasciolosis; Biochemical parameters; Snail; Permethrin; Cypermethrin; Plant extract. J Biol Earth Sci 201 3; 3(2): B1 64-B1 68 Corresponding author: J.N. Tiwari Department of Zoology, TD College Jaunpur, UP, India. E-mail:
[email protected] Original Submission: 11 May 201 3; Revised Submission: 1 0 June 201 3; Accepted: 1 7 June 201 3 Copyright © 201 3 Author(s). Journal of Biology and Earth Sciences © 201 3 Tomasz M. Karpiński. This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited. http://www.journals.tmkarpinski.com/index.php/jbes or http://jbes.strefa.pl e-mail:
[email protected] Journal of Biology and Earth Sciences, 201 3, Vol 3, Issue 2, B1 64-B1 68 B1 64 Six batches were prepared for each concentration. Advantages of natural product over synthetic have eco-friendly. free carbon dioxide and bicarbonate alkalinity were 6. These were identified at the herbarium of the Botany Department. precatorius were snail removed from 24h and 96h exposure period. Total free amino acids (µg/mg) were determined according to the method of Spies [1 0]. Extensive use of synthetic molluscicides are cause serious environmental hazards. Annual losses are estimated to be at least three million dollars [1 . Treated/control snails were removed from the glass aquaria and their gonadal tissue were dissected out. lakes and low-lying submerged area in Jaunpur. U.25±0. biodegradable abidance are less likely to accumulate in the environment. India. The snails were acclimatized for 72 hours in dechlorinated tap water. After DNA and RNA in gonadal tissue of L.2-6. Recently. A number of chemically diverse plant molluscicide has been used as molluscicides as and more effective than synthetic [7]. 6]. 24h the biochemical estimations were made in the gonadal tissue of the experimental snail. sativum and A. Homogenates (1 . 5. hatchability and survival of the young snails. DNA and RNA were exposing as mean ± SE. Student t-test was applied locate significance changes in between control and treated groups Sokal and Rholf [1 2]. Estimation of nucleic acids MATERIALS AND METHODS Test animals Adult snails L.Singh and Tiwari Influence of synthetic and plant extracts on gonadal tissue of Lymnaea acuminata INTRODUCTION Fasciolosis is an important helminth disease caused by two trematodes Fasciola hepatica and Fasciola gigantica [1 . cypermethrin. Plants used Plants used in this works were collected locally. In the present study describe the molluscicide of synthetic and extract of plants against certain biochemical changes in the gonadal tissue of snail Lymnaea acuminata. [8] reported that the synthetic molluscicides and extracts of plants caused a significant reduction in the fecundity. Permethrin and cypermethrin caused maximum Journal of Biology and Earth Sciences. A. Snails were exposed to sublethal concentrations (20% and 60% of 24h LC50) of different synthetic (Permethrin.20 cm in length) were locally collected from ponds. acuminata exposed to 20% and 60% of 24h LC50 of permethrin. Statistical analysis Each experiment was replicated at least six times. 2].57.0 mg/l. Singh and Tiwari..3 and dissolved oxygen. respectively. and 1 02. Issue 2. Fasciolosis is now recognized as an emerging human disease: The World Health Organization has estimated that 2. acuminata were estimated according to Schneider [11 ] using diphenylamine and orcinol reagents. acuminata. The fresh water snails Lymnaea acuminata and Indoplanorbis exustus are intermediate host of this parasite [3. Homogenates of gonadal tissue were prepared in 1 0% (w/v) trichloroacetic acid (TCA). DDU Gorakhpur University. protein. acuminata (2. 201 3. The supernatants were used for DNA and RNA estimations. Gorakhpur where voucher herbarium specimens (# No 3260 for Allium sativum and # 3829 for Abrus precatorius) are on deposit. B1 64-B1 68 B1 65 .P. The pH of the water was 7. Values of biochemical parameters protein. 4]. including humans.4–1 7 million people are infected with liverfluke.2. Cypermethrin) and Allium sativum bulb and Abrus precatorius fruit plants molluscicides.. respectively. amino acid. DNA and RNA level in the gonadal tissue of snail L. pools. 1 5-20 snails had to be dissected in order to obtain 1 00 mg nervous tissues. and a further 1 80 million are at risk of infection [5]. RESULTS There was a significant (p<0. amino acid. Control aquarium contained only dechlorinated tap water without treatment. DNA and RNA in gonadal tissue of L. The definitive host range of Fasciola is very broad and includes many herbivorous mammals.0 mg/ml. Vol 3.1 -7. amino acids. w/v) were prepared in 1 0% TCA at 90ºC and centrifuged at 5000 g for 20 minutes.3. Estimation of total protein and free amino acid Total protein (µg/mg) was estimated according to Lowry et al. Alternatively. [9] using bovine serum albumin as a standard.05) decrease in protein.0-1 05. the adherent tissue was removed and the organ was put on a filter paper for absorption of water. Different biochemical assay were performed viz. *Significant (p<0.Singh and Tiwari Influence of synthetic and plant extracts on gonadal tissue of Lymnaea acuminata Effect of 24h exposure to sublethal concentrations (20% and 60% of 24h LC50) of synthetic and plants molluscicides on the level of protein (µg/mg). amino acid (µg/mg). amino acid (µg/mg). Each value is mean ± SE of six replicates. Issue 2. Each value is mean ± SE of six replicates. *Significant(p<0. DNA and RNA (µg/mg) in the gonadal tissue of Lymnaea acuminata. Vol 3. Effect of 96h exposure to sublethal concentrations (20% and 60% of 24h LC50) of synthetic and plants molluscicides on the level of protein (µg/mg).05) when t-testwas applied in between treated and control groups. DNA and RNA (µg/mg) in the gonadal tissue of Lymnaea acuminata. Values of parentheses are per cent change with control taken as 1 00%.05) when t-test was applied in between treated and control groups. Journal of Biology and Earth Sciences. Table 2. Table 1. B1 64-B1 68 B1 66 . Values of parentheses are per cent change with control taken as 1 00%. 201 3. Agarwal RA. [1 4] have been reported that toxic effect of deltamethrin on the levels of biochemical changes in the snail Lymnaea acuminata. Report of the WHO informal meeting on use of trichlorobendazole in fasciolosis control.05) decrease was observed in protein. acuminata than the A. Whereas.96 % of control) (Table 1 and 2). precatorius caused a significantly change in certain biochemical parameters in the gonadal tissue of snail L. Mas-Coma S. 3. amino acid.92% of control) treatment with 60% of 24h LC50 of permethrin. 1 999. Amino acid levels in the gonadal tissue of the snail exposed to different preparation was significantly lower than control. 201 3. Bargues MD. acuminata exposed to 20% and 60% of 24h LC50 of A. precatorius. 1 988. Lymnaeids and human fascioliasis with a global overview on disease transmission. Singh A. S. The effect generally was more pronounced in the gonadal tissue. Rollinson. The synthesis of protein in any of a tissue can be affect in two ways by a chemical. 35: 1 255-1 278. It indicates that the biochemical changes in the gonadal tissue are altering the reproduction behaviour of these snails. In this case the RNA synthesis would be inhibited resulting in reduced RNA as well as protein content and only the protein content would be affected [1 3. 2. Molecular epidemiology and control.O. In conclusion. With the reduced RNA level there is bound to be a fall in protein levels because of reduction in the synthesis of new protein. whereas. 2009.H. Acta Hydrochim Hydrobiol. A. W. 1 7-1 8 October. Agarwal RA. acuminata. 5. A significant (p<0. Valero MA. 11 3-1 49.68 % and 7. The decrease in DNA and RNA content in L. Singh DK. Snail exposed to 20% and 60% of permethrin caused maximum reduction in total free amino acid (24h treatment -8. Iain Hay (Eds. acuminata may be because of cell death caused by release to toxic aldehydes resulting from peroxidation [1 7]. (Ed. 33. Molluscicide of plant origin.55 % of control) the level of protein. Journal of Biology and Earth Sciences. 2005. sativum and A.). cypermethrin and plant extract molluscicides with the protein synthesis. precatorius after exposure 24h and 96h. 7. Epidemiology. Advances in Parasitology. sativum and A. Geneva. 2009. Burlington: Academic Press. REFERENCES 1 .04 %. DISCUSSION It is evident from the results section that the sublethal exposure to 20% and 60% of 24h LC50 of permethrin. Zoonotic helminth infections with particular emphasis on fascioliasis and trematodiases. 1 4]. Mishra TN. 1 996. Valero MA. Vol 3. DNA and RNA level in the gonadal tissue of snail L. cypermethrin.Singh and Tiwari Influence of synthetic and plant extracts on gonadal tissue of Lymnaea acuminata reduction of protein. acuminata [1 6]. Epidemiology and Control. TRANSPARENCY DECLARATION The author declares no conflicts of interest. 1 6: 11 31 38. 364-376. The effect of pyrethroids and fenvalerate caused a significant inhibition in the protein level and enhancement in the amino acid level in the foot and hepatopancrease of snail L. amino acid. With the reduction in DNA levels change in RNA level is certainly not surprising. Mas-Coma S.49 % and 5. 96h treatment caused maximum reduction in (38.59 % of control). DNA and RNA level in the gonadal tissue of snail L. Fasciola. Int J Parasitol. Thus the treatments with 60% of 24h LC50 of permethrin and cypermethrin the both DNA and RNA levels decreased. In: D. 24h treatments with 20% and 60% of 24h LC50 of permethrin caused maximum reduction in protein level (40. B1 64-B1 68 B1 67 . 69: 1 41 -1 46.75 % and 36. Robinson MW. It indicates that they also interfere with the biosynthesis of amino acid in the cell [1 5]. Claxton J. CAB International. Phil Trans R Soc. 2006. Singh DK. WHO. acuminata.34 % of control) and (96h treatment -6. The result section has been demonstrated that the following treatments with permethrin and cypermethrin in levels of DNA and RNA decrease in a time and dose dependent manner.. pp. RNA level in gonadal tissue of snail L. Fasciolosis and other plant-borne trematode zoonosis.). Biol Agric Hortic. Issue 2. Singh et al. evolutionary genetics. (i) it either affect the RNA synthesis at the transcription stage or (ii) it some how affects the uptake of amino acid in the polypeptide chain. Reduction of protein levels may be due to direct interference of the permethrin. Harmful gastropods and their control.41 % of control). Bargues MD. Maximum reduction 24h in the level of DNA (27. acuminata was (1 7. In: Dalton JP. Switzerland. 6. Oxon. it can be stated that the component found in the synthetic (permethrin and cypermethrin) and extracts of plant alter the certain biochemical changes in snail L. 4. Dalton JP. Torgerson P. Journal of Biology and Earth Sciences.O. 1 3: 21 0-21 3. 1 5: 485-486. 8. Rosenbrough NJ. Kumar P. Press 1 957. Singh DK. Singh RN. WH Freeman. 11 . Singh A. Spies JR. Introduction to biostatistics. 1 6. Singh DK. Jahan M. 1 7. San Francisco. 1 957. 201 3. 1 999. Tiwari JN.) Methods in enzymology. Singh V. Singh VK. In: S. 1 2. Phytother Res. Sokal RR. B1 64-B1 68 B1 68 . Effect of binary combinations of deltamethrin + MGK 264 on the level of phospholipid and lipid peroxidation in the snail Lymnaea acuminata. 1 3. Schneider WC. Kaplan (eds. 9. Kumar P. 201 0. N. editors. Lowry OH. 1 951 . 1 977. Singh DK. Bull Environ Contam Toxicol. Colowick. In: Colowick SP. Determination of nucleic acid in the tissue by pentose analysis. Agarwal RA. pp: 464. Singh VK. Singh S. 201 2. Argemone mexicana and Nerium indicum on the certain biochemical parameters of Lymnaea acuminata. Singh DK. Protective effect of fruit extracts of Emblica officinalis (Gaertn) Terminalia bellerica (Roxb. Chemosphere. Effects of synthetic and extracts of plant on reproduction of snail Lymnaea acuminata. Toxic effect on deltamethrin on the levels of biochemical changes in the snail Lymnaea acuminata. Asif M. J Biol Chem. 1 993. 1 973. 3(8): 1 739-1 742. Hussain SJ. Tariq M. Singh VK. 1 0.P. 1 5. J Pharm Res. Singh RN. 1 93: 265-275. Ind J Exp Biol.) in experimental myocardial necrosis in rats. Issue 2. Res J Pharmac Biol Chem Sci. Methods in enzymology. Farr AL. Protein measurement with folin phenol reagent. Acad Press. 2008. 1 4. Kaplan NO. 1 4 New York: Acad. Effect of molluscicidal components of Abrus precatorius. 73: 1 032-1 035. 680. Vol 3. Effect of cypermethrin mexacarbate and phorate on phospholipids and lipid peroxidation in snail Lymnaea acuminata. Rohlf FJ. New York. 51 : 68-71 . Colorimetric procedure for amino acid.Singh and Tiwari Influence of synthetic and plant extracts on gonadal tissue of Lymnaea acuminata 1 3: 205-252. Randall RJ. 3(4): 705-71 0.