The Laboratory. Here's A Summary of Defined, Complex, Selective and Differential Media

Article Summary: Many types of bacterial growth media are used to culture (grow) microbes in
the laboratory. Here's a summary of defined, complex, selective and differential media.
Like captive animals in a zoo, bacteria grown in a laboratory environment need to have
everything provided for themfood, water, a suitable environmentin order to survive and
thrive.
Some microbes are not especially choosy in their requirements for growth, while others, such as
Treponema pallidum, the causative agent of syphilis, has never been successfully grown in
culture, although scientists have been trying to do so for more than 100 years.
Several basic types of media are discussed below. Although their differences are featured, there
are several characteristics that all culture media have in common:
Media must be prepared in such a way that it is sterile prior to being inoculated
with a bacterial sample, so that when a particular type of bacteria is cultured
(cultivated) on that medium, it is the only type of bacteria present.

Growth media must also provide everything the bacterial culture needs to live and
grow, including water, nutrients, and the proper pH. Media can be either liquid
(nutrient broth) or solid (agar).
Defined Media versus Complex Media

Some media formulations are very specific recipes in which certain ingredients must be present
in specific amounts. These defined media (also known as synthetic media) are used to grow
bacteria that have very particular needs.
Most clinical cultures do not have such exacting requirements, and can be grown in what is
referred to as complex media. Complex media are composed of partially digested yeast, beef, soy
and additional proteins, in which the exact concentration and composition is unknown. In
comparison with defined media, which are good for growing bacteria with very particular needs,
complex media can be thought of as a crowd-pleaser, suitable for growing many different types
of less fastidious microbes.
Tryptic Soy Agar (TSY) is a good all-purpose medium commonly used to grow bacteria in the
microbiology laboratory.
In addition to growth media formulations being classified as either defined or complex, there are
also specialized media that are designed to do more than just grow bacteria, selective and
differential media provide information about the bacteria growing.
Selective Bacterial Growth Media
For example MacConkeys Agar (MAC) is used to cultivate Gram-negative bacteria, by
discouraging the growth of Gram-positive bacteria through the use of crystal violet dyes and bile
salts. Another selective medium, Mannitol Salt Agar (MSA), has a high concentration of sodium
chloride, which selects for halophiles (salt-loving bacteria) such as members of the genus
Staphylococcus.
Differential Growth Media
Differential culture media are formulated to display a color change when the bacteria growing
metabolize a certain ingredient.
MacConkeys Agar, in addition to being selective (only growing Gram-negative bacteria), this
medium contains the sugar lactose and a pH sensitive dye.
When bacteria growing on MAC ferment lactose (metabolize it for food), they generate acidic
waste products that trigger the pH sensitive dye to turn the bacteria pink.
So, when grown on MAC, colonies of Gram-negative, lactose fermenting bacteria are pink, the
intensity of the pink color corresponds to how good the bacteria are at eating lactose. Colonies of
Gram-negative non-lactose fermenting bacteria grow in colorless colonies.
MacConkey's Agar growing Gram-negative bacteria. Salmonella, which doesn't ferment lactose,
is growing colorlessly in section #4. E. coli, which does ferment lactose, is growing pink in
section #3
Mannitol Salt Agar is another specialized differential media that contains food (mannitol, a sugar
alcohol) and a pH sensitive dye. When the bacteria growing on MSA ferment mannitol, the
medium changes from its original pink color to a bright highlighter yellow.
Bacteria that grow on Mannitol Salt Agar are all salt-loving halophiles. The salt is what makes
this medium selective. Mannitol Salt also contains the sugar-alcohol mannitol (food) and a pH
sensitive dye. This is the differential aspect of the medium.
If bacteria that cannot eat mannitol are growing on MSA (such as the normal flora species
Staphylococcus epidermidis) the medium remains pink. If halophilic bacteria that are able to eat
mannitol are growing, (such as the pathogen Staphylococcus aureus), the pH sensitive dye is
triggered by bacterial waste and the medium changes to bright yellow.
Another specialized medium, Blood Agar (BAP) contains sheeps blood. If bacteria growing on
the medium produce exotoxins that hemolyze (cut up) the red blood cells, the medium changes
color. BAP medium that has changed from red to transparent (completely clear) indicates betahemolysis, and possibly the pathogen Staphylococcus pyogenes. BAP that is bruised (alphahemolysis) or unaffected by bacterial growth present (gamma hemolysis) indicate normal flora,
non-pathogenic bacteria.

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Article Summary: Many types of bacterial growth media are used to culture (grow) microbes in

(cultivated) on that medium, it is the only type of bacteria present.

Defined Media versus Complex Media

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