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Microbial Cultivation
Microbial Cultivation
General microbial media. For the cultivation of bacteria, a commonly used medium
isnutrient broth, a liquid containing proteins, salts, and growth enhancers that will support
many bacteria. To solidify the medium, an agent such as agar is added. Agar is a
polysaccharide that adds no nutrients to a medium, but merely solidifies it. The medium
that results is nutrient agar.
Many media for microorganisms are complex, reflecting the growth requirements of the
microorganisms. For instance, most fungi require extra carbohydrate and an acidic
environment for optimal growth. The medium employed for these organisms is potato
dextrose agar, also known as Sabouraud dextrose agar. For protozoa, liquid media are
generally required, and for rickettsiae and viruses, living tissue cells must be provided for
best cultivation.
For anaerobic microorganisms, the atmosphere must be oxygen free. To eliminate the
oxygen, the culture media can be placed within containers where carbon dioxide and
hydrogen gas are generated and oxygen is removed from the atmosphere. Commercially
available products achieve these conditions. Anaerobic chambers can also be used within
closed compartments, and technicians can manipulate culture media within these chambers.
To encourage carbon dioxide formation, a candle can be burned to use up oxygen and
replace it with carbon dioxide.
To preserve microbial cultures, they may be placed in the refrigerator to slow down the
metabolism taking place. Two other methods are deep-freezing and freeze-drying. For
deep-freezing, the microorganisms are placed in a liquid and frozen quickly at temperatures
below –50°C. Freeze-drying (lyophilization) is performed in an apparatus that uses a
vacuum to draw water off after the microbial suspension has been frozen. The culture
resembles a powder, and the microorganisms can be preserved for long periods in this
condition.
Two processes for isolating bacteria from a mixed culture. (a) The
streak plate technique. (b) The pour plate technique.
A second isolation method is the pour plate method. In this method, a sample of bacteria
is diluted in several tubes of melted medium such as nutrient agar. After dilution, the
melted agar is poured into separate Petri dishes and allowed to harden. Since the bacteria
have been diluted in the various tubes, the plates will contain various dilutions of bacteria,
and where the bacteria are most diluted, they will form isolated colonies (Figure 1 ).