Culturing of Bacteria and Culture Methods
Culturing of Bacteria and Culture Methods
Culturing of Bacteria and Culture Methods
Cultivation/Culturing of Bacteria
• A microbial culture, is a method of
multiplying microorganisms by letting them
reproduce in predetermined culture
media under controlled laboratory conditions.
Uses:
Enrich the number of bacteria.
Select for certain bacteria and suppress others.
Differentiate among different kinds of bacteria.
Pure culture
• In the laboratory bacteria are isolated and
grown in pure culture in order to study the
functions of a particular specie.
• A pure culture is a population of cells or
growing in the absence of other species or
types. A pure culture may originate from a
single cell or single organism, in which case
the cells are genetic clones of one another.
• Pure cultures are obtained by using variety of special
techniques. All glassware, media and instruments
must be sterilized i.e. aseptic techniques are used for
obtaining pure cultures.
• Basic requirement for obtaining a pure culture are
solid medium, a media container that can be
maintained in an aseptic condition and a method to
separate individual cell.
• A single bacterium, supplied with right nutrients, will
multiply on the solid medium in a limited area to
form a colony, which is a mass of cells all descended
from the original one.
Agar
• Agar, a polysaccharide extracted from marine algae,
is used to solidify a specific nutrient solution.
• Unlike other gelling agent, it is not easily degraded
by many bacteria.
• It is not easily destroyed at higher temperatures, and
therefore it can be sterilized by heating, the process
which also liquefies it.
• Once solidified, agar medium will remain solid until
B. Solid media: An agar plate is a Petri dish that contains a growth medium
(typically agar plus nutrients) used to culture microorganisms. 2% of
agar is added. Agar is the most commonly used solidifying agent. Colony
morphology, pigmentation, hemolysis can be appreciated. Examples
include Nutrient agar and Blood agar.
C. Semi-solid media: Such media are fairly soft and are useful in
demonstrating bacterial motility and separating motile from non-motile
strains. Examples of Semi-solid media (Hugh & Leifson’s oxidation
fermentation). 0.5% agar is added.
Classification based on Nutritional
Components
1. Simple media.
2. Complex media.
3. Synthetic or chemically defined media.
Classification based on Nutritional Components
1. Simple media: Simple media such as peptone water, nutrient agar can
support most non-fastidious bacteria. It is also called as basal media. Eg:
NB, NA. Nutrient Broth consists of peptone, yeast extract and NaCl.
When 2% of agar is added to Nutrient Broth it forms Nutrient agar.
2. Complex media. Media other than basal media are called complex
media. They have special ingredients in them for the growth of
microorganisms. These special ingredients like yeast extracts or casein
hydrolysate, which consists of a mixture of many chemicals in an
unknown proportion.
1. Enriched media.
2. Selective media.
3. Differential media.
4. Transport media.
5. Indicator media.
6. Anaerobic media.
Classification based on Functional Use or Application
1. Enriched media :
• Addition of extra nutrients in the form blood, serum, egg yolk
etc to basal medium makes them enriched media.
• Media used to isolate pathogens from a mixed culture.
• Stimulate growth of desired bacterium and inhibit growth of
unwanted bacterium
• Media is incorporated with inhibitory substances to suppress
the unwanted organism, thus increase in numbers of desired
bacteria.
Examples of Enriched media: Chocolate agar Blood agar.
• Selenite F Broth – for the isolation of Salmonella, Shigella.
• Tetrathionate Broth – inhibit coliforms .
• Alkaline Peptone Water – for Vibrio cholerae.
• Chocolate Agar • Chocolate agar - is a non-selective,
enriched growth medium used for growing fastidious
bacteria, such as Haemophilus influenzae .
Uses:
Provides a pure growth
of bacterium for slide
agglutination and other
diagnostic tests.
Stab Culture
Prepared by puncturing a suitable medium –
gelatin or glucose agar with a long, straight,
charged wire.
Uses
– Demonstration of gelatin liquefaction.
– Oxygen requirements of the bacterium under
study.
– Maintenance of stock cultures.
Pour Plate Culture