Culture Media Microbio

Culture Media
Culture Media
Microorganisms need nutrients and
certain environmental conditions in order
to grow and reproduce. In the
environment, these microorganisms have
adopted to be habitats most suitable to
their needs. In the laboratory, however,
these requirement must be met by a
culture medium.
A culture medium is basically an
aqueous solution to which all the
necessary nutrients have been added.
Depending on the type, physical
properties, and combination of nutrients,
different classifications of culture media
can be made.
Culture Media
Culture media can be classified
according to three primary levels:
• Physical State
• Chemical Composition
• Functional Type
Physical State
Liquid Media
These are water-based
solutions that do not solidify at
temperatures above freezing
point. These media are
commonly formed broths, milk
and infusions
Semisolid Media
These exhibit a cloth like
consistency at ordinary
room temperature. These
media contain an amount of
solidifying agent ( agar or
gelatin) which thickens them
but does no produce a firm
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substance.
Solid Media
• These provide a firm surface
on which cells can form
discrete colonies and are
advantageous for culturing
and isolating bacteria and
fungi.
Solid media come in Two Forms:
1. Liquaefiable or 2. Non-liquefiable
Reversible Solid or Nonreversible
Media solid Media
Chemical
Composition
1. Synthetic
2. Non-synthetic
Synthetic
These compositions are chemically
defined. Synthetic media ae composed of
pure organic and inorganic compounds
which have molecular contents specified by
means of an exact formula.
Non-synthetic
These complex media contain at least one
ingredient that is not chemically defined, not a
simple compound, pure compound and not
representable by an exact chemical formula.
Most of these substances are extract of
animals, plants or yeast.
Example :
MacConkey Agar (MAC)
Is a widely-used culture medium which is
both selective and differential. The medium
is primarily used to differentiate between
Gram negative bacteria while inhibiting the
growth of most Gram positive bacteria.
Addition to the nutrient agar base of bile
salts and crystal violet will inhibit the
growth of most Gram positive bacteria,
making MacConkey agar selective.
Eosin Methylene Blue (EMB)
● Is a selective and differential
medium used for isolation and
differentiation among members of
the Enterobacteriaceae. Eosin
methylene blue agar selects for
Gram negative bacteria. It
contains methylene blue and eosin
dyes to inhibit the growth of
Gram positive bacteria.
Mannitol Salt Agar (MSA)
Was developed by microbiologists
seeking a method for isolating
Staphylococcus aureus, a pathogen
frequently transmitted by
contaminated food. This medium
contains 7.5% salt, which is inhibitory
to most bacteria other than
Staphylococcus.
Thayer-martin Agar
is a chocolate agar
designed to isolate
Neisseria gonorrhoeae.
Sabouraud’s Dextrose Agar (SDA)
is used for fungi. It contains

gentamicin and has a low pH
that will kill most bacteria.
Hektoen enteric Agar
is a selective medium with bile salts

added to inhibit Gram positive
organisms. The comparatively high
concentration of bile salts inhibits not
only Gram positive, but also some
Gram negative organisms, but not
Salmonella and Shigella species.
Sabouraud’s Dextrose Agar (SDA)
is used to isolate
Mycobacterium.
Mueller-Hinton Agar (MHA)
Is used to isolate Neisseria

gonorrhea and N. meningitidis
and also for anti-microbial
sensitivity testing.
Cetrimide Agar
is used to isolate Pseudomonas

aeruginosa. Salmonella-
Shigella Agar (SSA) is used to
isolate Salmonella and Shigella
species.
4. Differential Media
These allow growth of several
types of microorganisms and are
designed to display visible
differences among those
microorganisms. Differentiation
shows up as variations in colony
size or in color, in media color
changes or in the formation of
gas bubbles and precipitates.
MacConkey Agar (MAC)
Differentiates between lactose-fermenting coliforms
and lactose non-fermenters, which include potential
pathogens. Lactose, a fermentable carbohydrate, and
neutral red. a pH indicator, are added to differentiate
the lactose positive coliforms from the potentially
pathogenic lactose non-fermenters. When lactose is
fermented, acid products lower the pH below 6.8, with
the resulting colonial growth turning pinkish-red. If an
organism is unable to ferment lactose, the colonies will
be colorless.
Eosin Methylene Blue (EMB)
Differentiates those which ferment lactose (the
coliforms) from the coliforms which do not
ferment lactose. It contains lactose. Small
amounts of acid production result in a pink
colored growth, while large amounts of acid cause
the acid to precipitate on the colony, resulting in a
characteristic greenish, metallic sheen. Organisms
which do not ferment lactose will be colorless,
taking on the color of the medium. This medium
has been widely used in the past to screen for
coliforms in the water
Triple Sugar Iron Agar
Differentiates bacteria based on their
ability to ferment glucose, lactose and/or
sucrose, and to reduce sulfur to
hydrogen sulfide. It is used primarily to
distinguish the morphologically similar
bacteria of Enterobacteriaceae, all of
which ferment glucose to an acid end
product.
Mannitol Salt Agar
Is used to differentiate pathogenic
Staphylococcus species from nonpathogenic
members of the genus Micrococcus. If the
organism does not use mannitol, the medium
will remain red (no change). If the organism
does ferment mannitol, it will create
metabolic by-products which are acidic-and
the surrounding medium will be yellow.
Salmonella-Shigella Agar (SSA)
is used to differentiate Salmonella and

Shigella species whether they are
lactose (red colonies) and non-lactose
fermenters (colorless colonies) as well
as hydrogen sulfide producers (black
color).

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Culture Media

is used for fungi. It contains

is a selective medium with bile salts

Is used to isolate Neisseria

is used to isolate Pseudomonas

is used to differentiate Salmonella and

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