COLONY MORPHOLOGY

A) While studying colonies on solid media, the following features are noted:

Size: diameter in millimeters

Shape: circular, irregular, pin-point(punctiform), filamentous, spindle

shaped etc.

Surface: smooth, wavy, rough, papillate, glistening, dull etc.

Edge: entire, undulate, crenated, fimbriate, or curled.

Elevation: flat, raised, convex, umbonate.

Emulsifiability: whether the colony is emulsifiable or not.

Color: white, yellow, black, pink etc.

Consistency: butyrous, viscid, membranous, brittle, friable etc.

Change in medium: change in the color of the medium, liquefaction of

medium etc.

Density: opaque, translucent, transparent etc.

B) While studying growth in liquid media, the following features are noted:

Degree of growth.

Presence of turbidity and its nature.

Presence of deposit and its character.

Nature of the surface growth such as pellicle.

Pigment production if any.

C) Colony characteristics used in bacterial identification.

1) Hemolysis on Blood Agar
a) Alpha: Partial clearing of blood around colonies with green discoloration
of the medium; outline of red blood cells intact. e.g.: Streptococcus
pneumoniae, Viridans group of Streptococci
b) Beta: Zone of complete clearing of blood around colonies owing to lysis of
the red blood cells. e.g.: Staphylococcus aureus, Streptococcus pyogenes.
c) Gamma: No change in the medium around the colony; no lysis or
discoloration of the red blood cells. e.g.: Enterococcus species.
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2) Pigment production in agar medium

a) Water-soluble pigments discoloring the medium e.g.: Pseudomonas
aeruginosa
b) Non-diffusible pigments confined to the colonies e.g.: Staphylococcus
aureus, Serratia marcescens.

3) Changes in Differential Media suggestive of fermentation reaction

Various dyes, pH indicators are included in differential plating media to
serve as indicators of enzymatic activities and aid in identifying bacterial
isolates.

Medium
MacConkey

Sugar
Lactose

pH Indicator

Fermentation +ve Fermentation ve

Neutral red

Pink e.g.:

Pale e.g.: Proteus

Escherichia coli,

spp, Shigella spp,

Klebsiella

Salmonella spp.

Agar

pneumonia

Deoxycholate Lactose
Citrate Agar

Neutral red

Should be pink.

Pale e.g.: Proteus

Usually Lactose

spp, Shigella spp,

fermenting

Salmonella spp.

bacteria are
inhibited in this
medium due to
the high content
of Deoxycholate

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Xylose

Xylose,

Lysine

Lactose,

Phenol Red

Yellow e.g.:

Red e.g.: Shigella

Escherichia coli

spp (do not

Deoxycholate Sucrose

(ferment Lactose

ferment any of

Agar

& Sucrose. Acid

the sugars),

produced cannot

Salmonella spp.

be neutralized by

(ferment Xylose

the amines

only. But the

released from the

acid produced is

decarboxylation of

neutralized as it

lysine );

decarboxylates

Citrobacter &

lysine)

Proteus (ferment
Xylose only. The
acid produced is
not neutralized as
they do not
decarboxylate
lysine).

Thiosulphate

Sucrose

Citrate Bile

Bromothymol Yellow e.g.: Vibrio

Green e.g.: Vibrio

Blue

parahaemolyticus

cholerae

Sucrose
(TCBS) Agar

4) H2S production:
Certain organisms produce H2S which imparts blackening of the medium.
The media which help in detecting H2S contains certain indicators such as
ferric ammonium citrate/ferric citrate/ferrous sulphate which detect H2S by
imparting black color to the colony.eg: Salmonella typhi, Proteus spp.

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D) Characteristic Colony Morphology of Commonly Isolated Bacteria.

Bacteria

Media

Colony characteristics

Staphylococcus aureus

Blood Agar

Convex, entire edge, 2-3 mm,

golden/ creamy glistening
colonies with zone of beta
hemolysis.

Streptococcus pyogenes

Blood Agar

Convex or pulvinate,
translucent, circular,
pinpoint colonies with wide
zone of beta hemolysis.

Streptococcus pneumoniae

Blood Agar

Umbilicate or flat, translucent,

butyrous or mucoid colonies
with zone of alpha hemolysis

Escherichia coli

MacConkey Agar

Large, pink, circular, smooth

to rough, moist to dry, flat
colonies.

Blood Agar

Large, circular, greyish white,

moist, smooth opaque or
translucent colonies.

Klebsiella pneumoniae

MacConkey Agar

Large, pink, circular, smooth,

mucoid, convex colonies.
Large, circular, greyish white,

Blood Agar

mucoid, smooth, opaque

colonies.

Salmonella spp.

MacConkey Agar

Small, pale, circular, convex,

smooth colonies.
Small, red, convex colonies
with black centers due to H2S

XLD Agar

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production.

Shigella spp.

MacConkey Agar

Small, pale, circular, convex,

smooth colonies.
Small, red, circular, convex,

XLD Agar

smooth colonies without black

centers.

Proteus spp.

Blood Agar

Flat, gray, irregular,

spreading as thin film
(swarming growth) over agar
surface, fishy smell.
Flat, irregular, pale, non-

MacConkey Agar

spreading (no swarming)

colonies.

Pseudomonas aeruginosa

Nutrient Agar

Flat, greenish colonies having

effuse margins, with pigment
diffusing into the medium
Large, flat, irregular, pale

Vibrio cholerae

MacConkey Agar

colonies with effuse margins

TCBS Agar

Large, circular, convex, moist,

smooth, yellow colonies.

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