Culture Media for the Growth of Bacteria

For any bacterium to be propagated for any purpose it is necessary to provide the appropriate biochemical
and biophysical environment. The biochemical (nutritional) environment is made available as a culture
medium, and depending upon the special needs of particular bacteria a large variety and types of culture
media have been developed with different purposes and uses.

Culture media are employed in the isolation and maintenance of pure cultures of bacteria and are also
used for identification of bacteria according to their biochemical and physiological properties.

Types of Culture Media

Culture media may be classified into several categories depending on their composition or use.

Non-synthetic or complex medium

A complex (undefined) medium is one in which the exact chemical constitution of the medium is not known.
It contains

 a carbon source such as glucose for bacterial growth

 water
 various salts needed for bacterial growth
 a source of amino acids and nitrogen (eg beef extract and yeast extract )

Complex media usually provide the full range of growth factors that may be required by an organism so
they may be more handily used to cultivate unknown bacteria or bacteria whose nutritional requirement are
complex (i.e., organisms that require many growth factors )

Simple non-synthetic medium

A simple non-synthetic medium is capable of meeting the nutrient requirements of organisms requiring
relatively few growth factors.

Complex non-synthetic medium

A complex non-synthetic medium supplies more growth factors.

Table shows a Complex Medium for the Growth of Heterotrophic Bacteria

Constituent Amount
Peptone (partially digested protein) 5.0 g
Beef extract 3.0 g
Sodium chloride 8.0 g
Agar 15.0 g
Water 1 liter

Synthetic (Chemically defined) medium

A synthetic (defined) medium is one in which the exact chemical composition is known.
Chemically-defined media are of value in studying the minimal nutritional requirements of microorganisms,
for enrichment cultures, and for a wide variety of physiological studies.

Table shows a chemically defined Medium for growing a typical chemoheterotroph such as Escherichia

coli

Constituent Amount
Glucose 5g
Ammonium phosphate, monobasic (NH2H2PO4) 1g
Sodium chloride (NaCI) 5g
Magnesium sulfate (MgSO4. 7H20) 0.2 g
Potassium phosphate, dibasic (K2HPO4) 1.0 g
Water 1 liter

Pre-prepared medium

Media of many types can be obtained premixed, in an often dehydrated and powdered state. Such media
are called preprepared media.

Such media can be especially desirable if one lacks an autoclave, filtration devices etc

Reducing medium

Reducing medium is employed for growing obligate anaerobes. Reducing medium particularly contains
chemicals (reducing agents) such as sodium thioglycolate that chemically combine with dissolved oxygen
and deplete the oxygen in the culture medium.

Minimal media

Minimal media are those that contain the minimum nutrients possible for colony growth, generally without
the presence of amino acids, and are often used by microbiologists and geneticists to grow "wild type"
microorganisms.

Minimal medium typically contains

 a carbon source for bacterial growth, which may be a sugar such as glucose, or a less energy-rich
source like succinate
 various salts, which may vary among bacteria species and growing conditions; these generally
provide essential elements such as magnesium, nitrogen, phosphorus, and sulfur to allow the
bacteria to synthesize protein and nucleic acid
 water

Supplementary minimal media are a type of minimal media that also contains a single selected agent,
usually an amino acid or a sugar. This supplementation allows for the culturing of auxotrophs.

Selective media
A selective medium is one which has a component(s) added to it which will inhibit or prevent the growth of
certain types or species of bacteria to promote the growth of desired species.

It can also possible to adjust the physical conditions of a culture medium, such as pH and temperature, to
render it selective for organisms that are able to grow under these certain conditions. . For example, if a
microorganism is resistant to a certain antibiotic, such as ampicillin or tetracycline, then that antibiotic can
be added to the medium in order to prevent other microbes from growing which do not possess the
resistance.

Some examples of selective media include:

 Eosin-methylene blue agar (EMB) that contains methylene blue – toxic to Gram-positive bacteria,
allowing only the growth of Gram negative bacteria
 YM (yeast and mold) which has a low pH, deterring bacterial growth
 Blood agar (used in strep tests), which contains bovine heart blood that becomes transparent in
the presence of hemolytic Streptococcus
 MacConkey agar contains bile salts and crystal violet which inhabit the gram positive bacteria ,a
llowing only the growth of Gram-negative bacteria
 Mannitol salt agar (MSA) which is selective for Gram-positive bacteria
 Terrific Broth (TB) is used with glycerol in cultivating recombinant strains of Escherichia coli.
 Xylose lysine deoxycholate (XLD), which is selective for Gram-negative bacteria
 Buffered charcoal yeast extract agar, which is selective for certain gram-negative bacteria,
especially Legionella pneumophila

Differential media

Differential medium is one that distinguishes one microorganism type from another growing on the same
media. This type of media uses the biochemical characteristics of a microorganism growing in the presence
of specific nutrients or indicators (such as neutral red, phenol red, eosin y, or methylene blue) added to the
medium to visibly indicate the defining characteristics of a microorganism. This type of media is used for
the detection of microorganisms and by molecular biologists to detect recombinant strains of bacteria.

Examples of differential media include:

 Eosin methylene blue (EMB), which is differential for lactose and sucrose fermentation
 MacConkey (MCK), which is differential for lactose fermentation
 Mannitol salt agar (MSA), which is differential for mannitol fermentation
 X-gal plates, which are differential for lac operon mutants

Transport media

These are used for the temporary storage of specimens being transported to the laboratory for cultivation .
Such media ideally maintain the viability of all organisms in the specimen without altering their
concentration. Transport media typically contain only buffers and salt. -lack of carbon, nitrogen, and organic
growth factors prevents microbial multiplication. Transport media used in the isolation of anaerobes must
be free of molecular oxygen.

Example:

 Thioglycollate broth for strict anaerobes.

 Cary-Blair medium for preserving enteric pathogens
 Amies transport medium for ensuring the viability of gonococci

Enriched media

Enriched medium is designed to increase the numbers of desired microorganisms to a detectable level
without stimulating the rest of the bacterial population prior to plating on solid selective medium. Unlike
selective media, enrichment culture is typically used as broth medium.

Blood agar is an enriched medium in which nutritionally rich whole blood supplements the basic nutrients.
Chocolate agar is enriched with heat-treated blood (40-45°C), which turns brown and gives the medium the
color for which it is named.