Laboratory Exercise 8

CULTURAL CHARACTERIZATION OF BACTERIA

Microbes (not just bacteria) require appropriate conditions of incubation. These may include the
correct nutrients, chemicals, temperatures, etc. When the conditions are right, microbes are able to
multiply and will form colonies. A colony is the only way to see microorganisms with the unaided eye
and it should be remembered that colonies are large groups of microbes, not individuals. Using
nutrients, chemical, pH, temperature, salinity, and other conditions of incubation, a microbiologist is
provided with the first clue to what a particular organism might be.
When grown on a variety of media, bacteria will exhibit differences in macroscopic growth.
These differences, called cultural characteristics, are used as the basis of separating microorganisms
into taxonomic groups. The cultural characteristics for all known organisms are contained in Bergey’s
Manual of Systematic Bacteriology.
For accurate reporting, the microbiology student must learn the descriptive vocabulary of the
microbiologist. Descriptions of colonies must be concise and be phrased in such a manner that other
scientists will know what is being described. These descriptions are very important in the identification
and classification of the bacteria.

Objectives: At the end of the experiment, the student must be able to:
1. Observe the growth patterns of cultured microorganisms; and
2. Classify the growth behaviour of these microorganisms

Materials: Results of the culture medium in exercise 7 after incubation.

Procedure:
1. Bring your assignment on the cultural characteristics of the bacteria. Attach this copy on your lab
report.
2. Look closely at the grown cultures with which you have been working.
a. EMB plate
b. NB tubes
c. NA slants
3. Classify the colonies in your plate according to: size, form, elevation, margin, pigmentation, and
optical features.
4. Classify your slants according to: amount, form, consistency, and color.
5. Classify your broth medium according to: surface growth, subsurface growth, amount, and
sediment. (flocculation, turbidity, sediment or pellicle)

Agar Plate
These demonstrate well-isolated colonies and are evaluated in the following manner:
1. Size: Pinpoint, small, moderate or large.
2. Pigmentation: Color of colony
3. Form: The shape of the colony is described as follows:
a. Circular – Unbroken, peripheral edge
b. Irregular – Indented, peripheral edge
c. Rhizoid – Root-like spreading growth
4. Margin: The appearance of the outer edge of the colony is described as follows:
a. Entire – Sharply defined, even
b. Lobate – Marked indentations
c. Undulate – Wavy indentations
d. Serrate – Tooth-like appearance
e. Filamentous – Threadlike, spreading edge
5. Elevation: The degree to which the colony growth is raised on the agar surface is described as
follows:
a. Flat – Elevation not discernable
b. Raised – Slightly elevated
c. Convex – Dome-shaped elevation
d. Umbonate – Raised with elevated convex central region
Tube Culture (Slant)
These have single straight line of inoculation on the surface and are evaluated in the following manner:
1. Abundance of growth: The amount of growth is designated as none, slight, moderate, or large.
2. Pigmentation: Chromogenic microorganisms may produce intracellular pigments that produce
extracellular soluble pigments. that are excreted into the medium, that also produce a color. Most
organisms however, are non-chromogenic and will appear white to gray.
3. Optical characteristics: Optical characteristics may be evaluated on the basis of the amount of light
transmitted through the growth. These characteristics are described as opaque (no light
transmission), translucent (partial transmission), or transparent (full transmission).
4. Form: The appearance of a single line streak of growth on the ahar surface is designated as:
a. Filiform – Continuous, threadlike growth with smooth edges.
b. Echinulate – Continuous, threadlike growth with irregular edges.
c. Beaded – Noncofluent to semicofluent colonies
d. Effuse – This, spreading growth
e. Arborescent – Tree-like growth
f. Rhizoid – Rootlike growth

Tube Culture (Broth)

These are evaluated as to the distribution and appearance of the growth as follows:
1. Uniform fine turbidity – finely dispersed growth throughout
2. Flocculent – Flaky aggregates dispersed throughout
3. Pellicle – Thick pad-like growth on the surface
4. Sediment – Concentration of growth at the bottom of broth culture may be granular, flaky or
flocculent.

Tube Culture (Butt)

This solid medium may be liquefied by the enzymatic action of gelatinase. This liquefaction occurs in a
variety of patterns:
1. Crateriform – liquefied surface is saucer-shaped.
2. Napiform – Bulbous-shaped liquefaction
3. Infundibuliform – funnel-shaped
4. Saccate – Elongated, tubular
5. Stratiform – Complete liquefaction of the upper half of the medium
6. Motify
7. Gas-production
NAME: DATE PERFORMED:
Section:
Group number:

Laboratory Exercise No. 8

CULTURAL CHARACTERIZATION OF BACTERIA

RESULTS:
(In paragraph form, describe your observations and provide any illustrations of your results. Results
must be written in past tense and in 3rd person.)
Be sure to indicate what test microorganism was used!

DISCUSSION:
Based on the criteria given for evaluation of well-isolated colonies, state and/or discuss if your
experimental output met the criteria

Limit your results and discussion to 2-3 pages! Don’t forget to cite your references (as well as in-text
citations).