Criando uma coluna de Winogradsky: um método para enriquecer as espécies microbianas em uma amostra de sedimento
View the full transcript and gain access to JoVE Science Education videos
Q1: Why can't most Earth microorganisms be cultured in a laboratory?
Most microorganisms cannot be cultured in labs because they depend on other microbes within their native communities for metabolic products and survival. A Winogradsky column solves this by creating a miniature, enclosed ecosystem that mimics natural habitats and allows microbes to interact as they would in their original environment, enabling scientists to study them without isolation.
Q2: What is microbial succession and how does it occur in a Winogradsky column?
Microbial succession refers to the consecutive development of different microbial communities over time. In a Winogradsky column, microbes consume specific substrates and change the environment's chemistry. When substrates deplete, original microbes die off and new microbes with different metabolic needs flourish, creating visibly distinct layers with different microenvironmental requirements.
Q3: How do oxygen and sulfide gradients form in a Winogradsky column?
Photosynthetic cyanobacteria at the column's top produce oxygen, creating high concentrations near the surface and low concentrations toward the bottom. Simultaneously, sulfate reducers in anaerobic layers produce sulfide, which diffuses upward. These opposing gradients create distinct zones where different bacterial communities thrive based on their metabolic requirements for oxygen and sulfide.
Q4: What do different colored layers indicate in a Winogradsky column?
Green or red-brown layers indicate cyanobacteria; black sediment indicates sulfate-reducing bacteria; purple and green layers indicate photosynthetic sulfur oxidizers; white filaments indicate nonphotosynthetic sulfur oxidizers; and red-orange layers indicate purple non-sulfur bacteria. Each color represents distinct bacterial communities adapted to specific oxygen and sulfide concentrations within their microenvironment.
Q5: What substrates are added to a Winogradsky column and why?
Carbon sources, typically cellulose from shredded newspaper, and sulfur sources, usually from egg yolk, are added to the column. These substrates support microbial growth and metabolism. Optional additions include salt to enrich halophilic bacteria or a nail to enrich iron-oxidizing bacteria, allowing scientists to selectively cultivate specific microbial communities based on experimental goals.
Q6: How can environmental conditions be varied to enrich different microbial communities?
Light conditions can be modified by placing columns in high light, low light, or darkness, or by covering them with colored cellophane to select for specific bacterial groups. Temperature variations, such as incubating near a radiator for thermophilic bacteria or in a refrigerator for psychrophilic bacteria, also enrich different communities. These modifications allow researchers to study diverse microbial groups involved in biogeochemical processes.
Q7: What is the relationship between a Winogradsky column and enrichment cultures?
A Winogradsky column functions as a specialized enrichment culture that selectively grows microbial communities from sediment samples by mimicking natural habitats and creating chemical gradients. Unlike traditional enrichment cultures culturing aerobic and anaerobic microbes on selective and differential medias in isolation, the Winogradsky column preserves microbial interactions and community dynamics while enriching specific functional groups.