Winogradsky column

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This picture depicts the initial appearance of three different Winogradsky Columns. They are soil and water samples from a river, the later two columns have been modified with phosphate, nitrate and sulfur additives. These additions promote the growth of various bacteria specific to the anaerobic and aerobic regions of the column.

Shown above as a result of a 7 week period where the columns have been allowed to grow algae, cyanobacteria and other bacterial colonies. Of specific interest are the red regions of the middle column, indicative of purple non-sulfur bacteria(i.e Rhodospirillaceae) Also in column three, the red growth along the side of the column: a purple sulfur bacterium,Chromatium.

the green sulfur. place it in a sunny window. Mixed in with some shredded newspaper or hay (for cellulose). Add a few shredded cellulose fibers or paper. lake. beggiatoa. Invented by Sergei Winogradsky. Again. omitting any sticks.50% w/w calcium sulfate or sodium sulfate is required. depending on additive nutrients. a device based on a more elaborate one made by Russian microbiologist Sergei N.The Winogradsky Column is a simple device for culturing a large diversity of microorganisms. The column provides numerous gradients. ubiquitous in nature. Calcium sulfate of hard boiled egg yolk (as a sulfur source) Shredded paper (a piece of newsprint about 100 cm2) or cellulose fibers (as a carbon source) Procedure Mix the sand or mud with about a tablespoonful of egg yolk or calcium sulfate. rhodomicrobium. These two gradients promote the growth of different micro-organisms such as clostridium. desulfovibrio. Upper level high school students can use sophisticated sampling techniques of microorganisms and chemicals within the columns for both quantitative and qualitative studies. as well as many other species of bacteria. the purple sulfur. 1988). ground egg-shell and egg yolk respectively are rich in these minerals. First and second year high school students can do microscopic sampling of their communities that will introduce them to diversity among microorganisms. especially from a marshy area where colorful mats and scums of microorganisms can be found. students will use easily obtained materials to study ecological succession in a microbiological community. which adapted to the Earth's ancient environment by using sunlight. and a rich culture of photosynthetic bacteria . Pour the water you have collected over the top of the sand-mud mixture so that there is a small amount of unabsorbed water on top. Saltwater or freshwater marshes can be used. cyanobacteria. such as a stream. Photosynthesis occurred first in anaerobic bacteria. It is possible to establish a culture of photosynthetic bacteria in a "Winogradsky column". water. brings the container to two thirds full. Cover the bottle with plastic film secured by a rubber band. Due to low oxygen solubility in water the water quickly becomes anoxic towards the interphase of the mud and water. This is followed by water from the pond to saturate the mud and occupy half the remaining volume. or puddle. Elementary students will be fascinated by the changes occurring over time in their bottles. the device is a column of pond mud and water mixed with a carbon source such as newspaper (containing cellulose) or egg-shells (containing calcium carbonate) and a sulfur source such as gypsum (calcium sulfate) or egg-yolk. It is easier to fill the bottle if you first remove the narrow neck of the bottle by carefully cutting through the plastic with a single-edged razor blade or paper cutter. Layer the sand and mud in the plastic bottle. use either fresh water or salt water. the purple nonsulfur bacteria. At least four groups of photosynthetic bacteria abound on Earth today. The aerobic water phase and anaerobic mud or soil phase are one such distinction. Construction The column is a rough mixture of ingredients and exact measurements are not critical. Winogradsky. and the brownish nonsulfur Heliobacterium (Sagan and Margulis. from which the variety of aforementioned organisms can grow. Anaerobic phototrophs are still present to a large extent in the mud phase. The column is sealed tightly to prevent evaporation of water and incubated for several months in strong natural light. Green growth is often attributed to these organisms. and readily available carbon to produce carbohydrates and free oxygen. Supplementation of ~0. chromatium. >5cm wide) is filled by a third with pond mud. Materials Clear plastic 1 liter bottle Mud and sand from outdoor source. Background Bacteria. An additional anaerobic layer. External links Animated tutorial by Science Education Resource Center — Carleton College Winogradsky column: perpetual life in a tube — Edinburgh University Using A Winogradsky Column to Analyze Microbial Communities Frances Vandervoort 1991 Woodrow Wilson Biology Institute Objectives In this investigation. A tall glass or plastic tube (30cm long. as noted by the images. Water from outdoor source. Algae and other aerobic phototrophs are present along the surface and water of the upper half of the columns. this time of un-supplemented mud. and algae. are responsible for the evolution of the Earth's atmosphere.25% w/w calcium carbonate and ~0. chlorobium. debris or air bubbles. Incubating the column in sunlight for months results in an aerobic/anaerobic gradient as well as a sulfide gradient. The investigation is appropriate for a variety of age groups. there is still capacity for biofilm creation and colony expansion. pond. who lived in Russia and France the early part of this century.

Discussion Questions Describe the changes you see taking place over a period of several months. 239-240. Use a key to microorganisms to identify all you can. 1988. . How is a Winogradsky column similar to the bottom of a pond? How is it different? References Pigage. Boston: Harcourt. "The Winogradsky Column: A Miniature Pond Bottom. Pp. Record observations every one to two weeks. Helen K. Examine and draw those that you see. 1985. Jovanovich. Brace." American Biology Teacher (47/4). Watch the columns over several months. Garden of Microbial Delights. Dorion. April. Lynn. Using a long pipette. Sagan. Color changes indicate that ecological succession is taking place. A similar column kept in the dark will also develop a bacterial population.will develop within a few weeks. take samples from various depths within your column and place them on a microscope slide. and Margulis. but the bacteria will not be photosynthetic.


Presented below are some of the student results involving these columns. MA. except for column 8. and 2) model systems to study sediment-water biogeochemistry and microbial biogeochemical diversity.Results from the Winogradsky Column Study SES: Microbial Methods In Ecology. These columns served as 1) a source of microorganisms for labs on general microbial methods. Column 1 Column 2 . 8 in which freshwater sediments and water were used (see treatments). which was collected from a freshwater pond. and topped with estuarine water. except Col. such as bacterial productivity. Pictures were taken 6 weeks after initialization (9 Sep . Columns Columns were constructed from soda bottles (20 cm x 7 cm) and fitted with 5 luer lock sample ports and stopcocks (Cole-Parmer). students constructed Winogradsky columns from sandy sediments and water that were collected from Little Sippewisset estuary located in Falmouth. All columns (except 7) were filled approximately 3/4 full with sediments. Columns were placed in the laboratory near a SW-facing window.19 Oct 1999). 1999 In the Microbial Methods In Ecology Course.

Column 3 Column 4 Column 5 .

. Depth is measured from the lip of the column down.Column 6 Column 7 Column 8 Profiles of hydrogen sulfide (H2S) and methane (CH4) versus depth. Methane was measured on a gas Chromatogram and a spectrophotometric method was used to measure H2S concentration.

Column 1 Column 3 Column 4 .

Patchy formation of purple and green sulfur bacteria.Observations The following are some notable observations (click on images to obtain higher resolution picture): Formation of sulfur bacteria (probably Beggiatoa) in the top of column 2 (column that was placed in the dark). . and it is interesting that this column also exhibited low concentrations of H2S and CH4 as compared to the other columns. It is uncertain why this column developed such patches.

Only this column was amended with Fe (see Treatments). These anaerobic bacteria are phototrophic. The water in this column became anaerobic and stayed that way during all" . Purple and green sulfur bacteria are evident in several columns. The formation of iron sulfides (FeS) in Column 6 causes the sediments in this column to be black.Column 5. which received glucose. but use H2S as an electron donor instead of H2O. shown here. developed a hard mat or biopolymer at the air-water surface. Retrieved from "http://en. but are most pronounced in Column 7.

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