Micro Lab 5 - Colony Morphology

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Pure culture

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A culture that contains one type of microorganism.

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Subculture

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Microbes taken from one culture, are grown in a second culture to further purify it.

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Mixed culture

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Contains two or more identifiable, easily
differentiated microorganisms.

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Contaminated culture

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A culture that was once pure, but became mixed
by unwanted microbes of uncertain identity.

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Fastidious

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Microorganisms that require specific growth
factors and complex nutrients for growth.

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Quadrant streaking

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Method that dilutes and separates individual
microbial cells in order to produce
well‐isolated, pure colonies.

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Colony

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A visible population of microorganisms on solid
growth media, consisting of an aggregation
of microbes arising from a single parent cell.

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appearance/form,
color,
size,
margins/edges,
elevation,
and surface properties/texture.

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Colony characteristics: Characteristics include
overall

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Fastidious

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Requires specific growth factors.

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Complex (General Purpose) Media

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Used to grow the majority of bacteria,
it contains nutrients released by the partial
digestion of yeast, beef, soy, or other proteins;
its exact chemical composition is unknown.

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Chemically‐Defined (Synthetic) Media

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Used to grow fastidious bacteria that require
specific growth factors and nutrients;
its exact chemical composition is known.

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Selective Media

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Favors growth of specific microorganisms and
inhibits growth of unwanted ones.

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Differential Media

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Distinguishes between groups of microorganisms based on different chemical reactions and appearances.

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Enrichment Media

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Supplemented with blood, serum, or other growth factors to promote the
multiplication of specific microorganisms.

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Reducing Media

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Contains compounds that bind free oxygen
and remove it from the medium; used
to grow anaerobic microorganisms.

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Fermentation Media

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Contains carbohydrates that can be converted
to acids, plus a pH indicator to monitor acid
production (a by‐product of fermentation).

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Macroscopically,
microscopically,
biochemically,
immunologically,
and genetic analysis

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Five approaches for identifying a microorganism:

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(colony color, size, shape, color,
texture)

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Macroscopically

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(cell shape, size, motility, staining reactions)

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microscopically

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(enzymatic reactions)

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biochemically

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(antibody interactions)

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immunologically

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(DNA, RNA)

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and genetic analysis

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Quadrant streaking

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___________ dilutes the number of
colonies on plate and isolates colonies from
each other

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Aseptic technique

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___________ refers to the methods of
handling microbial cultures, patient specimens, and other sources of microbes in a way that prevents infection of the handler and others who may be exposed.

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Aseptic technique

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It prevents the introduction of unwanted microorganisms in a culture or environment.

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[1] Macroscopically (colony color, size, shape, color, texture); [2] microscopically (cell shape, size,
motility, staining reactions); [3] biochemically (enzymatic reactions); [4] immunologically (antibody
interactions); [5] genetic analysis (DNA, RNA).

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Name five approaches for identifying a
microorganism.

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Inoculation, incubation, isolation, inspection, and identification

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Name the “Five I’s” of bacterial culturing.

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Appearance/form, color, size, margins/edges, elevation, and surface properties/texture.

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Name six colony morphology characteristics.

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*Quadrant streaking dilutes the number of
colonies on plate and isolates colonies from
each other

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What is the purpose of quadrant streaking?

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Microbes often require *specific growth factors. *Fastidious bacteria require specific growth
factors/conditions and may be unable to
grow on general purpose media.

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Why can't all microorganisms grow on general
purpose media?

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At the *macroscopic level, after growth on solid
media you might observe *colonies that appear
different from a known microorganism. At the *microscopic level, after staining a sample you
might observe *cell shapes and *arrangements
that differ from the known microorganism.

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How can you tell if a culture has been
contaminated?

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*Aseptic technique refers to the methods of
handling microbial cultures, patient specimens, and other sources of microbes in a way that prevents infection of the handler and others who may be exposed. It prevents the introduction of unwanted microorganisms in a culture or environment.

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What is the purpose of using aseptic technique?

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[1] Using a sterile inoculation loop, obtain a loopful of test organism, and spread this sample back and forth in Quadrant I.
[2] Sterilize the loop, move the inoculation loop through the last few steaks of Quadrant I, and spread this sample in Quadrant II.
[3] Sterile the loop and spread the last few steaks of Quadrant II in Quadrant III.
[4] Sterile the loop and spread the last few steaks of Quadrant III in Quadrant IV, being careful not to overlap Quadrants I and IV.
[5] Sterile the loop, invert the plate, and incubate.

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Be prepared to describe and diagram the quadrant streaking technique.

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Colony Morphology

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Micro Lab 5 - is