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EXPERIMENT 2

SIMULATING THE ENVIRONMENT


WITH WINOGRADSKY COLUMN

Group 1
Aglibut
Ara ndia
Amisola
Balada d
Lantin

4B5

WINOGRADSKY COLUMN
By Sergei Nikolaevitch Winogradsky founder of soil
microbiology.
tool for microbial soil activity, nutrient cycling,
microbial succession and ecology.
Used to study carbon, nitrogen, sulfur, phosphorous
and etc.
2 gradients
A. Aerobic/ Anaerobic gradient
B. Sulfi de gradient
*promotes the growth of microorganisms

WINOGRADSKY COLUMN
Composition
A. Transparent cylinder
B. Marine or freshwater mud (diverse community of
interdependent microorganisms)
C. Substrates to support bacterial growth
* For six to eight weeks

Shredded newspaper source of carbon in cellulose form


Egg shell contains calcium carbonate
- yields carbon
Egg yolk source of sulfur

WINOGRADSKY COLUMN
Sulfi de diff uses upward
Oxygen diff uses downward
Sulfur oxidizing organisms
Consume both where they
meet
Stable counter balance of
sulfi de and oxygen
gradients
Allows organisms to grow
Figure 1

MATERIALS
Mud/soil sample

Pond water sample


2L clear soda bottle
1 hard boiled egg
Shredded newspaper
Iron nail/ screw
gallon empty ice
cream container

Parafilm/plastic/cling
wrap
Rubber band
Marker
Light source

METHODOLOGY
A. Winogradsky column
preparation
Slice neck of 2L soda bottle,
separate top from bottom column

Mark column in ml or cm

METHODOLOGY
B. Mud/ soil/ sediment sample Preparation
Mix 30g mud sample, 5g shredded newspaper,
pounded egg yolk, and pulverized egg shell, in an
empty ice cream container

Place mixture inside the soda bottle and add


pond water enough to cover the surface of the
mixture

METHODOLOGY
B. Mud/ soil/ sediment sample Preparation
Stir to release any trapped air bubbles in the soil

Add additional 500ml of pond water in the


mixture

Cover top with cling or plastic wrap with rubber


bands making sure that no air is introduced

METHODOLOGY
B. Mud/ soil/ sediment sample Preparation
Place set up near window where there is source
of natural/ artificial light

Observe weekly (for the next 6 weeks) record


data

RESULTS
WEEK 0

Figure 2

WEEK 1

Figure 3

WEEK 2

Figure 4

RESULTS
WEEK 3

Figure 5

WEEK 4

Figure 6

WEEK 5

Figure 7

RESULTS
Table
1
WEE
K

ODOR

COLOR
(of soil)

CONDENSATIO
N
(on plastic wrap
cover)

Crust
formation in
the bottle

Film on
water
surface

No foul
odor

Gray

None

None

None

Sulfuric
odor

Light gray

Present

Present

Present

Canal

Light gray
and green

Present

Present

Present

Canal

Light gray
and green

Present

Present

Present

Canal

Dark
green

Present

Present

Present

Canal

Black

Present

Present

Present

No data

No data

No data

No data

No data

DISCUSSION

Figure
8

http://archive.bio.ed.ac.uk/jdeacon/microbes/winograd
.htm

Cellulose
Initially from newspaper
Promotes rapid microbial growth
Depletes the oxygen in the water column and sediment
*Organisms that grow in anaerobic conditions are those that
ferment organic matter and can perform anaerobic respiration.
Fermentation
a process in which organic compounds are degraded
incompletely.
Anaerobic
a process in which organic substrates are completely
degraded to CO2.
uses a substance other than oxygen as the terminal electron
acceptor.

DISCUSSION
Cyanobacteria and Algae
need light to survive
the green coloring observed in the column on the illuminated side
Clostridium
strictly anaerobic (vegetative cells killed by oxygen exposure)
survive as spores in aerobic conditions
degrade cellulose to glucose which is fermented to gain energy and
produce simple organic compounds
Anaerobic Bacteria
contained egg yolk
enabled by the diffusion of H2S from sediment to water column,
dark green, purple or black colorings in the column
these colorings are groups of: green sulfur bacteria, purple sulfur
bacteria and sulfate-reducing bacteria
Will be further down the column

DISCUSSION
Sulfate reducing Bacteria
eat sulfur and make hydrogen sulfide gas
H2S is eaten by green and purple sulfur bacteria
Desulfovibrio
Purple Non-sulfur bacteria
need a carbon source to thrive
contained newspaper in the column
brown coloring

CONCLUSION
Different microorganisms can be observed on the Winogradsky
column and that the distribution of the aerobic and anaerobic
bacteria depend on the presence of oxygen and sunlight in the setup. Aerobic bacteria such as cyanobacteria and algae reside on
upper portion were oxygen and amount of sunlight is present while
anaerobic bacteria such as green sulfur bacteria, purple sulfur
bacteria and sulfate-reducing bacteria reside on the lower portion
were scarcity of oxygen and sunlight is observed . The varying
nutrient gradients and the presence of sulfur, nitrogen, carbon, and
phosphorus also contribute to the distribution of the different
bacteria in the Winogradsky column.