practical food microbiology Lab (2)

CULTURE MEDIA
A culture medium is any nutrient liquid or solid that can be used in the laboratory for
the growth of microorganisms.
Such a medium may resemble the natural substrate on which the microorganisms usually
grow (e.g. blood serum for animal pathogens, milk for milk microorganisms).

Whatever the medium, it must include all the necessary requirements for growth, which
vary according to the organism it is desired to grow but will include the following:-

A-Water
The role of water in the culture media is to dissolve all the medium components.
™ Distilled water is preferred in use, that is because the tap water may contain
calcium and magnesium salts which can react with phosphate found in
peptone or beef extract and produce calcium phosphate and magnesium
phosphate salts which are water insoluble which can inhibit the growth of
microorganisms.

B-Nitrogen containing compounds

(Peptides, proteins, amino acids, nitrogen-containing inorganic salts) in most cases
we use peptone as nitrogen source, and it has another function in the medium that is
buffering material. Because it is an amphoteric compound it can maintain the
medium pH.

C- Energy sources
(E.g. carbohydrates, peptides, amino acids and proteins) but in most cases we use
carbohydrates like glucose (dextrose).

D- Accessory growth factors

(Vitamins and mineral salts) usually we use beef extract as a source of the growth
factors.

E- Solidifying agents
These agents used only in the solid –reversible to liquid media. For example we use
gelatin or Agar agar as solidifying agents.

1. Gelatin
It is a protein which is added to the media in a concentration of 10-12 % of
the medium. It melts at 25-30o c and solidifies at below 25o c. If it is heated
to about 100o c or above it will melt but can not solidify again.

2. Agar agar
It is a carbohydrate which is added in a concentration of 1.5 - 2% to the
culture media.
At 98o C it is liquid and solidify at 42- 45o C.
Agar agar is the predominant solidifying agents, because:
™ Most bacterial strains can hydrolyze gelatin.
™ Gelatin can be molten at the incubation temperature.

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practical food microbiology Lab (2)

Classification of culture media

A- According to chemical composition:

™ Chemically defined media

(Contains defined components and we can reconstruct it again in the same
composition)

™ Non- chemically defined media

(Contain plant and animal tissues, and it has unknown composition so, we
can not reconstruct it. And usually it is a natural material like meat and
potatoes).

B- According to texture:

™ Liquid media
(Lactose broth)

™ Solid media
(Potato slices)
]
™ reversible to liquid
(Contain agar or gelatin as solidifying agents)

™ Semi solid media

(Contain 1/4 agar quantity of solid –reversible to liquid)

C- According to special purposes:

™ Selective media
Used to isolate one species of microorganisms and inhibit the growth of
the other microorganisms (for example, when we want to isolate G+
from a mixed culture of G+ and G- bacteria, we use a medium containing
crystal violet dye which inhibit the growth of G+ and not affect the
growth of G- bacteria.

™ Differential media
In which we add a natural or chemical compound to differentiate the
microbial groups which may found in the same culture.
Ex. Blood agar medium which contain blood to differentiate between the
blood hydrolyzing and non blood hydrolyzing bacteria, the blood
hydrolyzing bacteria which have colonies with clear zone around it.

™ Enriched media
In which we need to add blood serum or plant tissues to enrich the
growth of specific groups of microorganisms like heterophilic bacteria.