Winogradsky Column 2 6m4u4v

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 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 ½ 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

0

No foul odor

Gray

None

None

None

1

Sulfuric odor

Light gray

Present

Present

Present

2

Canal

Light gray and green

Present

Present

Present

3

Canal

Light gray and green

Present

Present

Present

4

Canal

Dark green

Present

Present

Present

5

Canal

Black

Present

Present

Present

6

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.