Winogradsky column
Device for culturing diverse microorganisms using gradients.
The Winogradsky column is a straightforward setup used to grow a wide range of microorganisms. It was created by Sergei Winogradsky in the 1880s. The device consists of a column filled with pond mud and water, supplemented with a carbon source like newspaper (which provides cellulose), eggshells (for calcium carbonate), and a sulfur source such as calcium sulfate or whole blended eggs. When left in sunlight for several months, the column develops both an aerobic-to-anaerobic gradient and a sulfide gradient. These gradients encourage the growth of many different microbes, including species like *Clostridium*, *Desulfovibrio*, *Chlorobium*, *Chromatium*, *Rhodomicrobium*, and *Beggiatoa*, along with various other bacteria, cyanobacteria, and even some eukaryotes.
The column supports multiple gradients, depending on the nutrients added, which allow these organisms to thrive. One key distinction is between the aerobic water phase and the anaerobic mud or soil phase. Because oxygen dissolves poorly in water, the water quickly becomes oxygen-depleted near the mud-water interface. Anaerobic phototrophs are abundant in the mud, where they can still form biofilms and expand colonies. Meanwhile, algae and other aerobic phototrophs live along the surface and in the upper half of the water column.
**Construction** Building the column involves a rough mix of ingredients—exact measurements are not crucial. A tall glass container (about 30 cm long and more than 5 cm wide) is filled one-third full with pond mud, with any sticks, debris, and air bubbles removed. To this mud, about 0.25% by weight of calcium carbonate and 0.50% by weight of calcium sulfate or sodium sulfate are added (ground eggshell and egg yolk are good sources of these minerals). Shredded newspaper, filter paper, or hay is also mixed in to provide cellulose. An additional layer of unsupplemented mud is then added, bringing the container to two-thirds full. Alternatively, some methods use sand for the layer above the enriched sediment to make it easier to observe and sample the resulting microbial populations. Pond water is then added to saturate the mud (or sand) and fill about half of the remaining volume. The column is sealed tightly to prevent water loss and incubated for several months in strong natural light.
Once sealed, anaerobic bacteria, including *Clostridium* species, develop first.
- Field
- Environmental microbiology
- Known for
- Inventing the Winogradsky column
Lore & Background
The Winogradsky column is constructed by filling a tall glass one third full of pond mud, supplemented with calcium carbonate and calcium sulfate or sodium sulfate, along with shredded newspaper or hay for cellulose. An additional layer of unsupplemented mud brings the container to two thirds full, followed by pond water. The column is sealed tightly and incubated for several months in strong natural light.
Reader's Guide
The Winogradsky column provides numerous gradients, depending on additive nutrients, from which a variety of organisms can grow. The aerobic water phase and anaerobic mud phase are one such distinction. Because of oxygen's low solubility in water, the water quickly becomes anoxic towards the interface of the mud and water. Anaerobic phototrophs are present in the mud phase, and algae and other aerobic phototrophs are present along the surface and water of the upper half. After the column is sealed, anaerobic bacteria such as Clostridium develop first, consuming cellulose and producing CO2. Eventually, color layers appear: a black anaerobic H2S dominated zone at the bottom, followed by green sulfur photosynthetic anaerobic bacteria, purple sulfur anaerobic bacteria, purple anaerobic non-sulfur bacteria, and at the top a layer of cyanobacteria that produce O2. While the column is an excellent tool to see whole communities of bacteria, it does not allow one to see densities or individual colonies, and it takes a long time to complete its cycle. Its importance in environmental microbiology should not be overlooked, and it remains an excellent tool to determine major bacterial communities before committing to molecular taxonomy methods such as shotgun sequencing.
Did You Know?
- The Winogradsky column was invented in the 1880s by Sergei Winogradsky.
- The column uses pond mud, water, a carbon source like newspaper, and a sulfur source like calcium sulfate or whole blended eggs.
- Incubating the column in sunlight for months results in aerobic/anaerobic and sulfide gradients.
- The column promotes growth of microorganisms such as Clostridium, Desulfovibrio, Chlorobium, Chromatium, Rhodomicrobium, and Beggiatoa.
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