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A Formal Report
Submitted to
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College of Science
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By
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4 Bio 5 Group 2
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March 2016
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Abstract
56a graduated cylinder or pre-calibrated empty soda bottle, filled with water, soil,
57and other substrates that would supplement nutrients and allow bacterial growth.
58The column is covered with saran wrap and rubber band to hinder evaporation of
59water. It is exposed to illumination to also allow phototrophic microorganisms to
60thrive. Shredded newspaper is used as primary source of carbon while hard61boiled egg is the primary source of sulfur. Two zones compose the Winogradsky
62column, the upper aerobic zone and lower anaerobic zone. The presence of an
63aerobic liquid air biofilm or pellicle may be seen on the surface. The column
64becomes more anaerobic at the lower layers. The objective of the experiment is
65to create a model microbial ecosystem that would show microbial activity and
66nutrient cycles of carbon and sulfur.
67Materials and Methods
68A 2-L clear, empty soda bottle was cut and marked from 0.0 to 100.0-mL with
69intervals of 5.0-mL. Egg yolk from a hard-boiled egg was pounded and the egg
70shell was pulverized. Shredded newspaper and soil/mud were mixed with the
71egg yolk and egg shell. The mixture was placed inside the calibrated soda bottle
72that will serve as the column. The bottle was filled with pond water until the
73surface of the mixture is covered by 3.0 to 4.0 cm. The mixture was stirred to
74release air bubbles and was left undisturbed for five minutes. More pond water
75was then added until the 500.0-mL was reached. The Winogradsky column was
76covered with plastic wrap and rubber band. Observations were done for four
77weeks and the while making sure that the water level was maintained by adding
78more pond water.
79Results and Discussion
80The end of the experiment left the column with few distinct layers: mold growing
81on top with black, white and red/ purple film, and green and white colored crust; a
82dark red/ brown layer; and a dark purple layer.
83The uppermost layer is consists of photosynthetic organisms including algae and
84cyanobacteria which, in presence of sunlight, these organisms photosynthesize
85to prepare their own food and oxygen is evolved in the reaction. The white film
86within and right below the water surface is the Microaerophillic zone where, even
87though one side is exposed to oxygen, very little O 2 diffuses slowly through water.
88This layer consists of bacteria such as Beggiatoa which oxidize the remaining
89Hydrogen-Sulfide into Sulfuric acid. The energy from which is then used to
90process other organic molecules. The upper portion of sand column is reddish
91brown
with
the
growth
of
non-sulfur
anaerobic
photoheterotrophs
92like Rhodospirillum. Below the red-brown zone, the red-purple zone indicates the
93growth of purple sulfur bacteria, Chromatium spp. and Ectothiorhodospirillum
94spp., which are photosynthetic organism. These organisms oxidize H2S into
95elemental sulfur (Shrestha, 2015).
96Decomposition and sulfate reduction activity of bacteria occur at the base of the
97column. 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. The different chemical demands each bacteria
100define the layer where they can thrive.
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Week 0
Rotten
Odors
egg
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
White Crust
crust
Rancid odor
Green and
white crust
Green Crust
Green, white
Green,
Green and
and
red/brown,
white film
red/brown
and white
film
film
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120Conclusion
121The Winogradsky column demonstrates metabolic diversity of microorganisms
122that grow in response to different environmental conditions. Different types of
123microorganisms
proliferated
and
occupied
distinct
zones
where
the
129References
130Brown,
J.
(2013).
Winogradsky
Columns.
Retrieved
131
http://www.mbio.ncsu.edu/mb452/Winogradsky_columns/wc.html
from
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133Shrestha,
T.
(2015).
The
Sufur
Cycle.
Retrieved
134
http://upendrats.blogspot.com/2015_03_01_archive.html
from
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136Winogradsky
Columns.
(n.d.).
Retrieved
from
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http://www.personal.psu.edu/faculty/j/e/jel5/biofilms/winogradsky.html
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