Culture media

“A culture medium is a solid or liquid mixture of chemicals that can support the growth of
microorganisms.”

Characteristic of ideal Culture Medium:

 Must give a satisfactory growth from a small inoculation.
 Should promote a rapid growth
 Should be easy to prepare
 Should be reasonably cheap
 Should be easily reproducible

Uses of Culture media:

 To identify cause of infection.
 To study the characteristics or properties of microorganisms.
 To prepare biological products such as vaccine.

Common ingredients/ composition of culture media:

 Peptone- source of carbon and nitrogen.
 Beef extract- source of amino acid, vitamins, minerals.
 Yeast extract- source of vitamin, carbon, nitrogen.
 Distilled water:
 Agar- solidifying agent.
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Classification of culture media:

BASED ON PHYSICAL NATURE:
1. Solid medium
2. Semi-solid medium
3. Liquid medium

1. Solid media
It is for the isolation of bacteria as a pure culture on a solid medium.
Agar is used to hardening the media at 1.5- 2.0% concentration. Solid media allows the growth of bacteria
as colonies by streaking on the medium. It solidified at 37 degrees Celsius.
Agar is an un-branched polysaccharide extracted from red algae species like Gelidium. Colonies
identification is done on this medium.

Examples of Solid Media

Nutrient agar, MacConkey agar, Blood agar, Chocolate agar.
Growth of bacteria on solid medium appear as smooth, rough, mucoid, round, irregular, filamentous,
punctiform.

2. Semi-solid media
This media shows the motility of bacteria and the cultivation of microaerophilic bacteria. This media has
agar at a concentration of 0.5% or less. It has a jelly consistency.

Examples of Semi-solid media

Mannitol motility media.
The growth of bacteria in semi-solid appears as a thick line in the medium.

3. Liquid media
This media shows the growth of a large number of bacteria.
It is called Broth that allows bacteria to grow uniformly with turbidity. The growth occurs at 37ºC in an
incubator for 24hrs.
Liquid media don’t have the addition of agar; it is for fermentation studies.

Examples of Liquid media

Nutrient broth, soy broth, phenol red carbohydrate broth.
Growth of bacteria in liquid media- Turbidity is seen at the end of the broth.

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Types of culture media based on chemical composition

1. Basal media or Simple media

2. Defined media or synthetic

3. Undefined or complex media

1. Basal media/ Simple media

This media is simple as it enhances the growth of many microorganisms. It’s a routinely used medium in
the lab, having Carbon and Nitrogen. This media allows the growth; of non- fastidious bacteria without
any enrichment source; used for sub-culturing. It’s a non-selective medium.

Staphylococcus and Enterobacteriaceae grow in this media.

Examples of Basal media

Nutrient Agar, Peptone water.

2. Defined medium or synthetic media

A defined medium has a known quantity of all ingredients, like carbon source (Glucose or Glycerol) and
nitrogen source (Ammonium salt or Nitrate as inorganic nitrogen). The medium needs in metabolic,
 nutritional, and physiological growth experiments. (Example: Simmons citrate agar, mineral glucose
medium.)

3. Undefined or complex media

This medium has different complex ingredients in unknown quantities, for example- yeast extract, beef,
various salts, and enzymatic protein. (Example: blood agar, milk agar)

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Types of culture media based on function

1. Enriched media
2. Selective media
3. Enrichment media
4. Indicator media or differential media
5. Transport media
6. Storage media

1. Enriched media
This media requires the addition of other substances like blood, egg, or serum. An enriched media allows
the growth of devised microorganisms but inhibits other and fastidious microbes grow as they require
nutrients like vitamins and growth-promoting substances.

Example of Enriched media

Blood agar, Chocolate agar, Lowenstein Jensen media.

2. Selective media
This media shows the growth of selective; microbes or desired microorganisms and inhibits the growth of
unwanted microbes. The inhibition occurs by adding bile salts, antibiotics, dyes, PH adjustments.

Examples of Selective media

S.N. Media Bacteria

1. Mannitol Agar– It has 7% of Selective for Staphylococcus aureus

sodium chloride that inhibits
the growth of other microbes
and promotes the growth
of Staphylococci. It has phenol
 red dye that produces acid
Staphylococcus used the
mannitol for the acid
production and the color of
phenol red changes from red to
yellow.
It is used for the isolation
Salmonella-Shigella Agar of Salmonella bacteria that causes
2.
Deoxycholate agar typhoid.
Selective for Shigella.
MacConkey Agar- It has bile
Selective isolation
3. salts that inhibit the growth of
for Enterobacteriaceae
gram-positive bacteria
TCBS Agar– Light green
translucent media Bile salt Selective for Vibrio cholera.
4.
inhibits the growth of unwanted
bacteria
Lowenstein Jensen Media- It is
made selective by adding
5. Selective for M. tuberculosis
malachite green and stops the
unwanted growth of pathogens.

3. Enrichment media
It is a liquid medium, which also permits the growth of desired bacteria at a low density. The media
provides an environment and conditions as selective media and inhibits unwanted bacteria from growing.
It is for the isolation of the soil and fecal microorganisms.

Examples of Enrichment media

Selenite F-broth does the isolation of Salmonella Typhi from a fecal sample.
4. Differential media or Indicator
This media shows visible changes due to biochemical properties or the presence of an indicator. It
differentiates bacteria based on colony color growing on the same plate.
Examples of Indicator or differential media
Mannitol salt agar shows yellow color colonies, MacConkey agar produces pink colonies.

5. Transport media
The media is use to transport specimens after collection to control the overgrowth of organisms. This
media act as temporary storage. It also maintains the viability of pathogens in the specimen and prevents
them from drying.

Examples of Transport media

Stuart’s transport medium. Pikes medium
6. Storage media
Media used for storing the bacteria for a long period of time. Examples are- cooked meat broth.
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Types of culture media based on oxygen requirement
Microorganisms have different requirements for growth depending on oxygen requirements.

1. Aerobic media
Aerobic media is the media that support the growth of bacteria that require oxygen to grow. Availability
of oxygen is through exposure to the air or by incorporating oxygen releasing components
Examples
Blood agar, Nutrient agar

2. Anaerobic media
Anaerobic media is the media that support growth of bacteria that don not require oxygen. Anaerobic
media contain reducing agent that help to remove oxygen.
Examples of Anaerobic media
RCM (Robertson cooked meat) isolation for Clostridium sp.
Thioglycolate broth– It has sodium glycolate that maintains low oxygen.

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