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COLLEGE OF SCIENCE
Activity 2
Winogradsky Column
Ecology Laboratory
BIO 203L
4 Biology 2
Group 1
18 April 2017
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UST College of Science Department of Biological Sciences
1 ABSTRACT
3 that illustrates the interdependent roles that prokaryotes play in sustaining life. In this
4 activity, the experimental set-up was composed of a mixture of newspaper, egg shell, egg
5 yolk, soil, and pond water in a transparent plastic bottle. Two set-ups were made, one
6 exposed to sunlight and the other covered with aluminium foil, to determine the function
7 of sunlight on the growth of microorganisms and different activities happening within the
8 column. The study was conducted for two months; wherein the set-ups were
9 photographed and observed twice every week. After two months, both set-ups showed
10 changes in color and odor, and exhibited various layers which indicated the separation of
11 the aerobic and anaerobic microorganisms into distinct zones that favour their specific
12 metabolic activities. Nutrient cycling was evident because of the obvious proliferation of
13 different microbes that required nutrients produced by other bacteria. This activity enabled
14 our group to create a microcosm in which complex microbial community processes affect
16 processes of how the carbon and sulfur cycles occur within a Winogradsky Column.
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UST College of Science Department of Biological Sciences
18 INTRODUCTION
19 Life is sustained by numerous cycles that are dependent on the taxonomic and
22 into various forms that can be used by other organisms. One such way to illustrate this in
23 the laboratory is with the use of the Winogradsky Column which was developed by two
25 understand the interdependent roles that various microorganisms play in order to sustain
28 driven by light. Using inexpensive materials, it creates conditions that demonstrate the
29 natural processes in which nutrients are cycled in the biosphere. Also, the Winogradsky
30 column is used to show the different aspects of how life was possible in early Earth which
33 community processes affect the surrounding environment was cultivated. Also, the
34 processes of how the carbon and sulfur cycles occur within a Winogradsky Column were
35 understood.
37 Sample collection. Approximately 300g of garden soil and 500mL of pond water were
38 collected from four different sites [(1) Bacoor, Cavite; (2) Holy Family Residence; (3)
39 Calauan, Laguna; (4) Tarlac] then combined to obtain integrated soil and water samples.
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40 Winogradsky column. The neck of two 2L transparent soda bottles were cut. The yolk
41 of one hardboiled egg was separated from the egg white then made into small crumbs.
42 The eggshells were pulverized into a fine powder. The egg yolk crumbs and eggshell
43 powder were mixed along with shredded newspaper and the integrated soil sample. The
44 soil mixture was divided into two and placed in the two cut soda bottles. The integrated
45 pond water sample was added to each soil mixture until it filled approximately of the
46 soda bottle. The open end of each soda bottle was covered with cling wrap. One column
47 was exposed to sunlight while the other was covered with aluminium foil. The columns
49 RESULTS
24 Jan
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31 Jan
3 Feb
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7 Feb
10 Feb
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14 Feb
21 Feb
28 Feb
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7 Mar
21 Mar
28 Mar
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31 Mar
51
Condensation
cover
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53 Table 2 cont.
Observa- Week 5 Week 6 Week 7
tions (Mar 7 & 10) (Mar 17 & 21) (Mar 28 & 31)
Condensation
cover
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Color of
Brown Brown Brown Brown Brown Black Black
soil
Conden-
sation on
Absent Absent Absent Absent Absent Present Present
plastic
cover
Thin
Crust Thin film
orange
forming in None None None None None (light
crust on
the bottle orange)
sides
56
57 Table 3 cont.
Observa- Week 5 Week 6 Week 7
tions (Mar 7 & 10) (Mar 17 & 21) (Mar 28 & 31)
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Conden-
plastic cover
58
59 DISCUSSION
62 diffuses downward from the surface, fermentation products and microbial metabolites
63 diffuse upward. The cycling of nutrients within the column creates various chemical
64 gradients that are necessary for the growth of certain organisms, enabling their
66 those present in natural ecosystems (Anderson & Hairston, 1999). In order for a
67 Winogradsky column to work, it initially requires a sulfur, inorganic carbon, and cellulose
68 source. In the experimental set-up, the egg yolk crumbs served as the sulfur source, the
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69 eggshell powder served as the inorganic carbon source, and the shredded newspaper
71 Growth was observed in the columns one week after it was prepared (Table 1).
72 Both set-ups turned into a dark colored mixture with visible green growths at the top. The
73 dark color of the mixture is attributed to rapid microbial growth promoted by the presence
74 of cellulose. The green growths observed at the top of the mixture indicate the growth of
76 growth of microorganisms depletes the oxygen present at the bottom of the set-up,
79 products, such as organic acids and alcohols, which diffuse upward. These fermentation
80 products together with the sulfur and inorganic carbon sources added into the mixture are
82 hydrogen sulfide.
83 By the fourth week, the soil-pond water mixture has a distinct odour similar to that
84 of rotten eggs because of the sulfur by-products of certain microorganisms. The columns
85 also showed different zones with distinct colors, starting from the bottom, the colors were
86 opaque black, green, red, and dark green with sparse orange-brown areas for the light
87 set-up, orange-brown with sparse dark green areas for the dark set-up. The different
88 colors at distinct zones present in the column indicate the type of microorganism that has
90 blackened areas, due to the formation of ferrous sulfide, in the lower portion of the column
91 where conditions are anaerobic. The sulfide products of Desulfovibrio are then used by
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93 acceptor, yielding elemental sulfur and water; its growth was indicated by the green zone
94 directly above the blackened area at the lower portion of the column. Above this zone,
95 was a red colored band which can be ascribed to the accumulation of non-sulfur bacteria
97 Rhodopseudomonas. These organisms are photoheterotrophs that trap light energy and
98 use organic molecules as both electron and carbon sources. The presence of this zone
99 in the dark set-up is attributed to fact that the foil covering the experimental set-up was
100 lost. Lastly, the combination of dark green and orange-brown areas at the topmost layer
101 in both the light and dark set-ups indicate the growth of photosynthetic cyanobacteria,
103 microbes are chemoautotrophs that oxidize hydrogen sulfide to sulfate to gain energy for
104 the synthesis of organic matter. The sulfate produced cycles back to the anaerobic
105 sediment of the column to be used by Clostridium, completing the sulfur cycle within the
107 The key factor that sustains life within the Winogradsky column is nutrient cycling.
108 In the column, carbon, hydrogen, and oxygen are cycled through aerobic respiration and
109 photosynthesis. On the other hand, sulfur, which is an important nutritional requirement
110 for most life, is cycled within the column through aerobic and anaerobic respiration. The
111 sulfur cycle is dependent on the elements chemical variability. Changes in the oxidation
112 states of sulfur are mediated by microbial metabolisms, which is perfectly illustrated within
113 the Winogradsky column. For instance, anaerobic species use elemental sulfur as the
114 terminal electron acceptor in respiration which reduces it into hydrogen sulfide. While
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115 others can use thiosulfate or sulfate as an electron receptor. Also, algae and many
117 (Anderson & Hairston, 1999; Deacon, 2003; Rogan et al., 2005).
118 CONCLUSION
120 cycles that occur within the biosphere. The interplay of various organisms in the cycling
121 of nutrients prove the importance of microbial metabolic diversity. This activity enabled
122 our group to create a microcosm in which complex microbial community processes affect
123 the surrounding environment. Also, it provided knowledge regarding the different
124 processes of how the carbon and sulfur cycles occur within a Winogradsky Column.
125 REFERENCES
126 Ackert, L. T. (2007). The cycle of life in ecology: Sergei Vinogradskiis soil microbiology,
128 Anderson, D. C. & Hairston, R. V. (1999). The Winogradsky column & biofilms: models
129 for teaching nutrient cycling & succession in an ecosystem. The American Biology
131 Deacon, J. (2003). The Microbial World: Winogradsky Column: perpetual life in a tube.
133 http://archive.bio.ed.ac.uk/jdeacon/microbes/winograd.htm
134 Rogan, B., Lemke, M., Levandowsky, M., & Gorrell, T. (2005). Exploring the sulfur nutrient
135 cycle using the Winogradsky column. The American Biology Teacher, 67(6), 348-
136 356.
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