Вы находитесь на странице: 1из 15

UST College of Science Department of Biological Sciences

UNIVERSITY OF SANTO TOMAS

COLLEGE OF SCIENCE

Activity 2

Winogradsky Column

Ecology Laboratory

BIO 203L

4 Biology 2

Group 1

Celina Joyce Aniceto

Tricia Anne Barot

Eleazar John Cruz

Rafaella Beatriz Kraft

18 April 2017

i
UST College of Science Department of Biological Sciences

1 ABSTRACT

2 The Winogradsky Column is an inexpensive and indispensable tool in microbial ecology

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

15 the surrounding environment. Also, it provided knowledge regarding the different

16 processes of how the carbon and sulfur cycles occur within a Winogradsky Column.

17 Keywords: Winogradsky Column, nutrient cycling, soil microbial ecology

1
UST College of Science Department of Biological Sciences

18 INTRODUCTION

19 Life is sustained by numerous cycles that are dependent on the taxonomic and

20 metabolic diversity of microorganisms. For instance, the metabolic diversity of

21 prokaryotes enables the sulfur cycle, an essential constituent of life, by transforming it

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

24 microbiologists, Sergius Winogradsky and Martinus Willem Beijerinck, who wanted to

25 understand the interdependent roles that various microorganisms play in order to sustain

26 life (Anderson & Hairston, 1999; Ackert, 2007).

27 The Winogradsky Column is a complete, self-contained recycling system that is

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

31 was dominated by sulfur-based, anaerobic microbes (Rogan et al., 2005).

32 In this activity, the ability to create a microcosm in which complex microbial

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.

36 MATERIALS AND METHODS

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.

2
UST College of Science Department of Biological Sciences

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

48 were observed and photographed twice a week for two months.

49 RESULTS

50 Table 1. Winogradsky column observations


Date Light Dark

24 Jan

3
UST College of Science Department of Biological Sciences

31 Jan

3 Feb

4
UST College of Science Department of Biological Sciences

7 Feb

10 Feb

5
UST College of Science Department of Biological Sciences

14 Feb

21 Feb

28 Feb

6
UST College of Science Department of Biological Sciences

7 Mar

21 Mar

28 Mar

7
UST College of Science Department of Biological Sciences

31 Mar

51

52 Table 2. Weekly observations of the Winogradsky column (light set-up)


Observa- Week 1 Week 2 Week 3 Week 4
tions (Jan 24 & 31) (Feb 3 & 10) (Feb 14 & 21) (Feb 28)
Odor None None Foul Foul Foul Foul Foul

Color of soil Brown Brown Black Black Black Black Black

Condensation

on plastic Absent Absent Present Present Present Present Present

cover

Orange Orange Orange

Thin film ring ring ring


Crust forming
None None None (light present present present
in the bottle
colored) at the at the at the

surface surface surface

8
UST College of Science Department of Biological Sciences

Green Green Green Green Green


Film on the
moss- moss- moss- moss- moss-
surface of the None None
like like like like like
water
band band band band band

53 Table 2 cont.
Observa- Week 5 Week 6 Week 7
tions (Mar 7 & 10) (Mar 17 & 21) (Mar 28 & 31)

Odor None None Foul Foul Foul Foul

Color of soil Black Black Black Black Black Black

Condensation

on plastic Present Present Present Present Present Present

cover

Orange Orange Orange Orange Orange Orange

ring ring ring ring ring ring


Crust forming
present present present present present present
in the bottle
at the at the at the at the at the at the

surface surface surface surface surface surface

Green Green Green Green


Film on the Green Green
moss- moss- moss- moss-
surface of the moss-like moss-like
like like like like
water band band
band band band band

54

9
UST College of Science Department of Biological Sciences

55 Table 3. Weekly observations of the Winogradsky column (dark set-up)


Observa- Week 1 Week 2 Week 3 Week 4
tions (Jan 24 & 31) (Feb 3 & 10) (Feb 14 & 21) (Feb 28)
Odor None None Foul Foul Foul Foul Foul

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

Mold-like Mold-like White White


Film on the
film on film on mold-like mold-like
surface of None None None
top with top with film on film on
the water
bubbles bubbles top top

56

57 Table 3 cont.
Observa- Week 5 Week 6 Week 7
tions (Mar 7 & 10) (Mar 17 & 21) (Mar 28 & 31)

Odor None None Foul Foul Foul Foul

Color of soil Black Black Black Black Black Black

10
UST College of Science Department of Biological Sciences

Conden-

sation on Present Present Present Present Present Present

plastic cover

Thin Thin Thin


Crust Orange Orange Orange
orange orange orange
forming in crust on crust on crust on
crust on crust on crust on
the bottle sides sides sides
sides sides sides

White White White White White White


Film on the
mold-like mold-like mold-like mold-like mold-like mold-like
surface of
film on film on film on film on film on film on
the water
top top top top top top

58

59 DISCUSSION

60 The Winogradsky Column is an inexpensive device used to study the different

61 functions of various microorganisms in nutrient cycling and sustaining life. As oxygen

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

65 proliferation in distinct zones and creating a vertical distribution of microbes similar to

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

11
UST College of Science Department of Biological Sciences

69 eggshell powder served as the inorganic carbon source, and the shredded newspaper

70 served as the cellulose source (Rogan et al., 2005).

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

75 green, photosynthetic microorganisms such as cyanobacteria and algae. The rapid

76 growth of microorganisms depletes the oxygen present at the bottom of the set-up,

77 creating an anoxic environment. At such conditions only anaerobic bacteria, such as

78 Clostridium, can survive. Anaerobes degrade cellulose and produce fermentation

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

81 utilized by anaerobic, sulfur-reducing organisms, such as Desulfovibrio, to produce

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

89 accumulated in that area. The development of Clostridium and Desulfovibrio appear as

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

12
UST College of Science Department of Biological Sciences

92 anaerobic photosynthetic bacteria such as Chlorobium to serve as its final electron

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

96 that are able to grow in microaerophilic conditions such as Rhodospirillum and

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,

102 green algae, and sulfur-oxidizing organisms such as Thiobacillus. Sulfur-oxidizing

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

106 closed system.

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

13
UST College of Science Department of Biological Sciences

115 others can use thiosulfate or sulfate as an electron receptor. Also, algae and many

116 heterotrophic microorganisms may utilize sulfate by incorporating it into proteins

117 (Anderson & Hairston, 1999; Deacon, 2003; Rogan et al., 2005).

118 CONCLUSION

119 The Winogradsky column is an inexpensive device used to illustrate complex

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,

127 1885-1940. Journal of the History of Biology, 40, 109-145.

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

130 Teacher, 61(6), 453-459.

131 Deacon, J. (2003). The Microbial World: Winogradsky Column: perpetual life in a tube.

132 Retrieved on 15 April 2017, from:

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.

14