THE CULTURAL CHARACTERISTICS OF MICROORGANISMS

What is a “microbiological culture”?

When bacteria or other microorganism grow on a laboratory medium, they are referred to as a
culture. Different species of bacteria growing on the same kind of medium may appear
different. Knowing the cultural characteristics of the species may aid in the identification of the
organism.

Microorganisms exist in nature as a mixed culture. However, to determine the characteristics

of a particular species, it is important that the organism be isolated and grown in the laboratory
as a pure culture (a culture containing only one species of organism).

What are the common methods of isolating microbial cultures?

There are several techniques of isolating pure culture. The most common are the streak-
plate/spread plate technique and pour plate method.

Streak Plate or Spread Plate Technique. The streak plate or spread plate is obtained
by transferring a portion (usually 0.1 mL) of a bacterial suspension on the surface of an agar
medium and streaking or spreading over the surface. Spread plates are usually streaked with a
sterile bent glass rod. This process thins out the bacteria on the agar surface so that individual
bacteria are separated from each other.

Pour Plate Technique. The pour plate technique involves the principle of dilution or
thinning out the concentration of the bacterial suspension. After a series of transferred into an
empty petri dish. Agar medium is poured into the culture plate, allowed to solidify and incubate.
Some organisms may be trapped beneath the surface of the medium when it gels. With this,
the pour plate exhibits both surface and sub-surface colonies.

What are the characteristics of colony formation?

One of the major features of the bacteria is their appearance or growth characteristics following
incubation in an appropriate medium. The growth characteristics of a pure culture is determined
by observing the following types of culture: (a) colonies on plate cultures, (b) growth on agar
slants, (c) growth in broth, and (d) growth on agar stabs.

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plate culture slant culture agar stab culture broth culture

Agar plate cultures are obtained by the pour plate technique or the spread plate technique.
Agar slant cultures are prepared by making a streak inoculation up the middle of the sloped
surface with a transfer needle. For the stab culture, the transfer needle carrying the inoculum is
introduced in a straight line from the top to the bottom of the tube and removed along the same
path. Tubes of broth can be inoculated with the transfer needle or loop.

The features of growth in different culture media are presented below:

Agar Plate Colonies

a. Size. Colonies range in size from extremely small (pinpoint) measuring only a fraction
of a millimeter in diameter, to large colonies measuring 5 to 10 mm in diameter.
b. Margin or Edge. The periphery of bacterial colonies makes many different patterns,
depending upon the species. It may be very evenly circular, like the edge of a droplet,
or it may show such irregularities as rounded projections, irregular notches, threadlike
projections or root-like projections.
c. Elevation. The colonies may be very thin (flat) to thick (raised). Raised colonies
exhibit varying degrees of a convex structure.
d. Chromogenesis. Colonies may be colored (pigmented) or not (nonpigmented).
Various shades of red, yellow, brown and violet characterized certain species.
e. Optical features. May be opaque, translucent, or opalescent.

The cultural characteristics of bacterial colonies on agar plates is summarized below:

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Agar Slant Growth

a. Amount. Scanty, moderate or abundant.

b. Margin or Edge of Growth. Uniform or even or exhibiting various irregularities similar
to the irregularities described or colonies.
c. Consistency of mass of growth. Butyrous or butter-like consistency; viscous or
stringy; dry and brittle.
d. Chromogenesis. Similar to that described for colonies.

The cultural characteristics of bacterial colonies on agar slants is presented below:

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Growth in Nutrient Broth

a. Amount of Growth. Scanty, moderate or abundant.

b. Distribution of growth throughout the broth. Uniformly distributed (evenly turbid);
growth confined to surface of broth as a scum or film (pellicle); growth accumulated
as sediment, which may be granular or viscous.
c. Odor. May be putrid, fruity or aromatic, or negligible.

Bacterial colonies in nutrient broth may appear as follows:

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Growth in Agar Stabs

a. Amount of Growth. Scanty, moderate or abundant.

b. Margin or Edge of Growth. Uniform or even or exhibiting various irregularities similar
to the irregularities described or colonies.
c. Chromogenesis. Similar to that described for colonies.

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YEAST COLONIES

Attempts have been made to utilize the characteristics of yeasts colonies to identify yeast
strains. Young colonies are ointment-like in consistency, while older ones are denser, tending
to dry out, and may form pigments which give the organism a characteristic color (yellow,
orange, pink, brown or black). The appearance of yeasts in liquid culture may also be used to
identify the organism. Some species grow on the bottom of the culture vessel forming a
sediment, whereas other species exhibit a uniform turbidity. Another group grows only at the
surface, forming a thicker layer.

Some examples of yeasts are presented below:

Candida alabicans Saccharomyces cerevisiae Hansenula

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MOLD COLONIES

The presence of molds in food is characterized by the fuzzy, cottony appearance that tend to
scatter on the surface as it grows. In general, the mold colony is described based on the color
of the mycelium and the extent at which the organism spread in the medium. Some examples
of mold culture are presented below:
Mold culture Aspergillus flavus
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Penicillium Alternaria Geotricum candidum

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Botrytis cinerea Rhizopus stolonifer

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Suggested Links:

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