12 3 Simulating the Environment With A Winogradsky Column 4 5 6 7 A Formal Report 8 Submitted to 9 Sonia N. Javier, M.Sc. & Jonathan Carlo A. Briones, M.Sc. 10 Department of Biological Sciences 11 College of Science 12 University of Santo Tomas 13 14 15 16 17 18 By 19 20 4 Bio 5 Group 2 21 Ysabel Aira E. Azucena 22 Judy Ann G. Bacud 23 Shannon Cake M. Baquiran 24 Timothy John L. Bautista 25 26 27 28 March 2016 47 48Keywords: Winogradsky column. sulfur. shredded newspaper. Different types of microorganisms such 41as such as Cyanobacteria and Clostridium proliferated and occupied distinct 42zones where the environmental conditions favour their specific activities. nitrogen. pulverized egg shell. sulfur. Nutrient cycling 49 50Introduction 51The Winogradsky column is a tool that was designed by Sergie N. 43Conclusion. 31Winogradsky to study microbial activity and succession and nutrient cycling of 32carbon. 53nitrogen. phosphorus. The objective of the 33experiment was to create a model microbial ecosystem that would show 34microbial activity and nutrient cycles of carbon and sulfur. The column consists of .29 Abstract 30Introduction. The Winogradsky column demonstrates metabolic diversity of 44microorganisms and illustrates how different microorganisms perform their 45interdependent roles wherein the activities of one organism enable another to 46grow. 35Method. soil. The set-up is ideal for studying 54growth of microorganisms because the materials needed to construct a 55Winogradsky column are inexpensive and easy to obtain. The Winogradsky column is a tool that was designed by Sergie N. An empty plastic container was filled with a mixture of pounded egg 36yolk. and other nutrients. The column was observed for four weeks for microbial 38activity and nutrient cycling 39Result. and other nutrients. Winogradsky 52to study microbial activity and succession and nutrient cycling of carbon. The bacterial growth responds in patterns or in distinct zones reflecting 40the substrate and aerobic concentrations. and pond water and 37covered with plastic wrap. microbial activity. phosphorus. The mixture was placed inside the calibrated soda bottle 72that will serve as the column.0-mL. 79Results and Discussion 80The end of the experiment left the column with few distinct layers: mold growing 81on top with black. 58The column is covered with saran wrap and rubber band to hinder evaporation of 59water. 57and other substrates that would supplement nutrients and allow bacterial growth. More pond water 75was then added until the 500. white and red/ purple film. and a dark purple layer.0 cm. filled with water. Shredded newspaper is used as primary source of carbon while hard61boiled egg is the primary source of sulfur. The Winogradsky column was 76covered with plastic wrap and rubber band.0-mL was reached.0 to 100. and green and white colored crust. Two zones compose the Winogradsky 62column. Egg yolk from a hard-boiled egg was pounded and the egg 70shell was pulverized. It is exposed to illumination to also allow phototrophic microorganisms to 60thrive. Shredded newspaper and soil/mud were mixed with the 71egg yolk and egg shell. empty soda bottle was cut and marked from 0. in presence of sunlight. a 82dark red/ brown layer.56a graduated cylinder or pre-calibrated empty soda bottle. The mixture was stirred to 74release air bubbles and was left undisturbed for five minutes. Observations were done for four 77weeks and the while making sure that the water level was maintained by adding 78more pond water. The presence of an 63aerobic liquid air biofilm or pellicle may be seen on the surface. 83The uppermost layer is consists of photosynthetic organisms including algae and 84cyanobacteria which. The objective of the experiment is 65to create a model microbial ecosystem that would show microbial activity and 66nutrient cycles of carbon and sulfur.0-mL with 69intervals of 5. these organisms photosynthesize .0 to 4. 67Materials and Methods 68A 2-L clear. soil. The bottle was filled with pond water until the 73surface of the mixture is covered by 3. the upper aerobic zone and lower anaerobic zone. The column 64becomes more anaerobic at the lower layers. Heterotrophic microorganisms also 108thrive inside the column. Lower portions of the Winogradsky column are often 106colonized by green photoautotrophic and purple sulfur bacteria. Examples of these are obligate anaerobes like clostridia 109and methanogenic bacteria as well. Metal sulfides like iron usually contribute to 110the black color at the bottom of the column. The breaking down of the materials that make up the substrates 98contributed to the production of different substances needed by the bacteria at 99different levels of stratification. 2015). 114 . The white film 86within and right below the water surface is the Microaerophillic zone where.85to prepare their own food and oxygen is evolved in the reaction. The aerobic layer 107is often occupied by oxygenic cyanobacteria. 88This layer consists of bacteria such as Beggiatoa which oxidize the remaining 89Hydrogen-Sulfide into Sulfuric acid. Below the red-brown zone. Sulfur oxidizing organisms consume 111oxygen that diffuse downward and sulfide that diffuse upward. 96Decomposition and sulfate reduction activity of bacteria occur at the base of the 97column. which are photosynthetic organism. Reduction of sulfate causes the 102release of hydrogen sulfide gas at the lower layers and decreases at the higher 103layers. These organisms oxidize H2S into 95elemental sulfur (Shrestha. This reaction 112results to a stable counterbalance of sulfide and oxygen and ultimately allows the 113growth of organisms with either oxygen or redox requirements. and Ectothiorhodospirillum 94spp. the red-purple zone indicates the 93growth of purple sulfur bacteria. Carbonate in the column also functions as pH buffer and is 105important of the autotrophs. The formation of a gradient allows a great range of habitats for various 104microorganisms. Chromatium spp. Anaerobic microorganisms cause the 101release of sugar by cellulose fermentation. even 87though one side is exposed to oxygen. The energy from which is then used to 90process other organic molecules.. The different chemical demands each bacteria 100define the layer where they can thrive. The upper portion of sand column is reddish 91brown with the growth of non-sulfur anaerobic photoheterotrophs 92like Rhodospirillum. very little O 2 diffuses slowly through water. 0 Weekly observation of the Winogradsky Column Observations Week 0 Rotten Odors egg Color of the soil odor Gray None Condensation on plastic cover Crust forming in the Bottle Film on Surface of the water None None Week 1 Week 2 Week 3 Week 4 Rancid odor Rancid odor Rancid odor Gray Gray Gray Gray colorless colorless colorless Colorless Green and Green and White White Crust crust Rancid odor Green and white crust Green Crust Green. white Green.0 Week 2 . Green and and red/brown. film film 116 117 Figure 1.115 Table 1. White and white film red/brown and white red/ purple film. Black.0 Week 1 Figure 2. 0 Week 3 Figure 4. and other substances that create the stratification of 127microbial growth. 128 . Different types of 123microorganisms proliferated and occupied distinct zones where the 124environmental conditions favour their specific activities such as Cyanobacteria 125and Clostridium.0 Week 4 120Conclusion 121The Winogradsky column demonstrates metabolic diversity of microorganisms 122that grow in response to different environmental conditions. oxygen. Decomposition of different substrates contributed to the diffusion 126of hydrogen sulfide.118 119 Figure 3. personal. Retrieved 134 http://upendrats. The Sufur Cycle.d.html from 135 136Winogradsky Columns.ncsu.).edu/mb452/Winogradsky_columns/wc. Retrieved from 137 http://www. (2013). (n. Winogradsky Columns.blogspot.html 138 .129References 130Brown. J.edu/faculty/j/e/jel5/biofilms/winogradsky.mbio.com/2015_03_01_archive.psu. (2015).html from 132 133Shrestha. T. Retrieved 131 http://www.