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Exercise 2: Simulating the Environment with a Winogradsky Column

J. J. Gaa1, A. P. Ramirez1, J. A. Raqueo1 4BIO2 (Group 6)


Department of Biological Sciences, College of Science, University of Santo Tomas, Espaa
Avenue, Manila

Abstract

shreddings, carbonate from the egg shell

The Winogradsky column is a very


useful tool in studying microbial activity and
the

cycling

of

nutrients

and

other

compounds between the aerobic zone and


the

anaerobic

zone.

Soil,

newspaper

shreddings, a hard-boiled egg and pond


water were used to create a microcosm of

and sulfate source from the egg yolk. The


column set-up has an anaerobic lower area
and

aerobic

upper

area

which

the

microorganisms or microbial populations


develop in the aspect that is related to the
concentration gradients of oxygen, sulfur,
nutrients, and light.

microbial communities and to see how

Microbes

grow

rich

on

different

carbon and sulfur cycles in an ecosystem.

nutritional therapy that allows microbes to

The set-up is left in a shaded area of the lab

recycle substances in nature which is

and the observations were recorded for six

important to eliminate waste products. Some

weeks.

of these microbes are aerobic that can grow

Keywords:

Aerobic

Respiration,

Respiration,

Microcosm,

Anaerobic

Nutrient

cycle,

Winogradsky column

in the presence of air while the others are


anaerobic which cannot grow if air is
present. The general idea of this experiment
is, as oxygen diffuses downward from the
surface

which

fermentation

Introduction

is

the

causes the

aerobic

zone,

breakdown

of

cellulose from the shredded newspaper and

The Winogradsky column was named

hydrogen sulfide diffuses upward from the

after the Russian microbiologist, Sergei

lower aerobic zone. The anaerobic sediment

Winogradsky,

at the bottom of the column will most likely

is

simple

laboratory

experiment used to exhibit how different

be

populated

microorgamisms execute their independent

Desulfovibrio species. Above the sediment

roles in a completely contained system of

there will be an inverse of gradients of H 2S

recycling. This Winogradsky column is made

and SO4 that will appear with the formation

from a clear, thin plastic or glass column

of

filled with saturated soil. The soil has been

sulfur and purple sulfur bacteria layers.

fortified with carbon from the newspaper

(Pevzner & Shelton, 2001)

the

by

anaerobic

Clostridium

and

photosynthesis, green

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The aim of this exercise is to create a

the components. Slowly pour the mixed

microcosm in which complex microbial

pond and aquarium water in the column until

communities processes affect the cultivated

3cm. Take off the cut out paper and let it

surrounding

an

settle for 5 minutes. Cover the column with a

appreciation for the diversity of methods

cling wrap and fasten it with rubber band

microorganism use to gain energy from

and masking tape. The created Winogradsky

oxygen-producing

column

bacterial

environment,

to

gain

photosynthesis

photosynthesis

and

lastly,

and
to

diagram, the carbon and sulfur cycles as it


occurs in a Winogradsky column.

set-up

is

exposed

at

room

temperature and placed in a sunny window.


The Winogradsky column set-up is
checked and recorded every week, for six
weeks. The observations were drawn from
the appearance of the column and any

Methodology

changes

to

the

column

including

any

To create the Winogradsky column, a

changes in color patterns, growth in the soil

2.0 L soda bottle was used as its base (or

and the soil-water interface were recorded

column) by slicing its neck with a pair of

and documented.

scissors or cutter. Calibrate the plastic


column from 0.0-500.0 ml with intervals of
100.0 ml. Prepare all of the components
needed to establish a microenvironment
inside the column.

Results and Discussion


The Winogradsky column set-up was
observed once a week for six weeks and

Weigh 90 g of soil and ~10 g of

notes the appearance of the column. The

newspaper shreddings. With the use of a

experiment started on January 29, 2016 and

mortar and pestle, pound an egg yolk until

was designated as week 0. During this

the lumps disappears and the egg shell from

period, the soil appeared to be brown in

an egg until it is almost pulverized. With a

color as well as the water.

large

the

was not present during this period as well as

newspaper

the crust and film on the surface of the

shredding, hard-boiled egg (not including its

water, were both absent. The coloration of

egg white) in an ice cream container.

the water and the soil may be due to the

spoon,

components

homogeneously
namely

soil,

mix

Cut a circle using a paper with the

Condensation

sediment not yet settling on the bottom.

same diameter of the soda bottle, this would


be

used

to

prevent

shreddings to rise

the

inside

newspaper
the

column.

Transfer the mixed components inside the

Table 1: Observation with the cover on (Shaded)

column and place the cut out paper on top of

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Figure 2: Winogradsky Column (Week 1)

Figure 1: Winogradsky column (Week 0)

The second and third week or Weeks


1 and 2, happened on February 5 and 12,

Figure 3: Winogradsky Column (Week 2)

respectively. The second week produced a


greenish-brown colored soil and the water
was clear. The third weeks soil color was
green-brown with two very distinct layers
with the dark brown soil on the very bottom
with a layer of shredded newspaper on top
of it. Both weeks showed condensation on

Weeks 3, 4, 5, and 6, which happened


on February 19, and 26, and on March 4
and 11, respectively, showed very distinct
layering throughout the set-up. A very thick
crust has formed on the surface and some
forms of fungi have been seen growing on
top of this crust.

the sides and on the plastic cover as well as


a presence of a crust and film on the waters
surface.

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Figure 4: Winogradsky Column (Week 3)

layer of this Winogradsky set-up is where


most of the solid materials ended up. This
layer has a black color showing this layer is
devoid of oxygen or an anaerobic zone. This
layer is where fermentation takes place and
in this layer, purple sulfur bacteria are found
which give the layer a red color and on the
very bottom layer of the set-up, sulfurreducing bacteria are found which gives the
bottom a black color (Rogan, et al., 2005).
On the upper part of the bottom layer, where
oxygen levels are depleted, it is where
anaerobic cellulose degraders are found
where they ferment the cellulose to glucose,
which in turn they use for energy (Rogen, et
al., 2005).

Figure 5: Winogradsky Column (Week 4)

Figure 7: Winogradsky Column (Week 6)

Conclusion
Figure 6: Winogradsky Column (Week 5)

Through this Winogradsky column setThe water shows two distinct layers, a
murky upper layer and a less murky lower
layer. The upper layer is home to oxygenproducing organisms or the aerobic zone.
The lower layer is where most of the
hydrogen sulfide (H2S) gas is diffusing
upward to the aerobic zone (Rogan, et al.,
2005).
The upper layer is home to
cyanobacteria and algae that undergo
photosynthesis thus giving this layer a green
color while the lower layer is where sulfuroxidizing bacteria are found. The bottom

up, we were able to observe the different


changes happening inside the column.
Different microbial organisms were seen
such as algae and fungi. Different types of
microorganisms proliferated and created
distinct zones. These zones are where the
environmental conditions favor the activities
of these microorganisms.

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The Winogradsky column is an


excellent way of determining and learning
about the complex microbial communities

Pevzner,

Y.,

&

Shelton,

S.

(2001).

and their roles in the cycling of nutrients.

Winogradsky Column Unit. Retrieved

This set-up demonstrates the diversity of

March

microorganisms present in an area and how


specific environmental conditions response
to their activities.

16,

2016,

from

https://www.westminstercollege.edu/edu
cation_gslp/YevgenyPevznerWinograds
kyColumn.pdf

References
Rogan, B.,
Deacon, J. (n.d.). The Microbial World:
Winogradsky column: Perpetual life in a
tube. Retrieved March 18, 2016, from
http://archive.bio.ed.ac.uk/jdeacon/micro

Lemke, M., Levandowsky, M.,

Gorrell, T. (2005). Exploring the sulfur


nutrient cycle: Using theWinogradsky
column. The

American

Teacher, 67(6),

348-356

Biology

bes/winograd.htm

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