[BIO 203 L] Exercise 2 Winogradsky (Preliminary)

Exercise 2: Simulating the Environment with a Winogradsky Column J. J. Gaña1, A. P. Ramirez1, J. A. Raqueño1 4BIO2 (Group 6) Department of Biological Sciences, College of Science, University of Santo Tomas, España 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

a

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 week’s 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 water’s 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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