Chapter 8

USE OF COLONY MORPHOLOGY FOR

PRESUMPTIVE IDENTIFICATION OF
MICROORGANISMS

Presenter:
Assoc. Prof. Lea D. Ballares,
RMT, M. Bio. Ed., MSMT
 Objectives:
After studying this chapter, you should be able to:
• Describe how growth on blood, chocolate, and Mac Conkey
agars is used in the preliminary identification of isolates.
• Differentiate a-hemolysis from B-hemolysis on blood agar
culture medium
• Associate the colony characteristics shown on blood,
chocolate, and Mac Conkey agars with microscopic findings
on direct smear, and use the information in the presumptive
identification of microorganisms.
COLONY - macroscopically COLONIAL MORPHOLOGY
visible growth of a microorganism - Colony characteristics and
on a solid culture medium form.
 A simple method of mechanically diluting
a relatively large concentration of
Streak Plate Technique microorganisms to a small, scattered
population of cells.
Initial observation of culture
• Observation of colony morphology
is done after 18 to 24 hours
incubation
• Factors affecting growth &
colony morphology:
– Culture medium
– Environmental conditions - oxygen
carbon dioxide, temperature, pH, and
moisture
(details discussed in chapter 6)
 PLATE READING – Comparative
examination of colony morphology of
Interpretation of Culture microorganisms growing on various
culture media.
Chocolate agar: most organisms and
fastidious organisms

Mac Conkey agar: gram- Blood agar: grows most

negative organisms organisms
Interpretation of Culture
• Differentiation using plated
media
– Lactose fermenters (pink)/
nonlactose fermenters
(colorless)
Colony Characteristics
 Size
 Form or
Margin
 Elevation
 Density
 Consistency
 Color
 Pigment
 Odor
Colony Characteristics
• SIZE – pinpoint/punctiform, small, medium, large

Gram - rods

Neissria gonorrhea

Gram + cocci

Bacillus cereus
Colony Characteristics
• FORM OR MARGIN- smooth, filamentous, rough or irregular

Examples:
• Bacillus anthracis is filamentous
• Diphtheroid colonies have rough edges

Bacillus anthracis Diptheroid

Colony Characteristics
 ELEVATION- raised, convex, flat, umbilicate, umbonate
Examples:
– Umbilicate: convex with depressed center
(pitting); S. Pneumoniae (if no capsule)
– Umbonate: convex with protruding nipple;
diphtheroids
Colony Characteristics
 DENSITY- translucent, transparent, opaque

B-Hemolytic streptococci Staphylococcus aureus

Colony Characteristics
 COLOR- white, gray, yellow or buff

Coagulase negative Staphylococci Neisseria spp.

Colony Characteristics
 CONSISTENCY- butyrous, creamy, brittle, mucoid

Staphylococcus aureus Nocardia spp.

Examples:
• S. aureus: creamy
• Neisseria: sticky
• Nocardia: brittle
• Streptococci: dry
• Diphtheroid: dry and waxy
Colony Characteristics

Lactose-fermenting Escherichia/Citrobacter-like organisms

growing on MAC

Klebsiella/Enterobacter-like lactose fermenters growing on

MAC
Colony Characteristics
 PIGMENT

Serratia marcescens Pseudomonas aeruginosa

EXAMPLES:
 Serratia marcescens: brick red  Chromobacterium violaceum: purple
 Pseudomonas aeruginosa: green  Prevotella melaninogenica: brown-
 Kluyvera spp: blue black pigment
Colony Characteristics
 ODOR

EXAMPLES:
• S. aureus: old sock
• P. aeruginosa: fruity or
grapelike
• Proteus mirabilis: putrid
• Haemophilus spp.: musty
basement
• Nocardia spp.: freshly plowed
field
Colony Characteristics HEMOLYSIS ON BAP

TYPES OF HEMOLYSIS:
• β hemolysis: complete clearing
of blood cells around the colonies

• α hemolysis: partial clearing of

blood that results in a green
discoloration of the medium
• Nonhemolytic colonies are
referred to as  hemolysis.
Colony Characteristics HEMOLYSIS ON BAP

Staphylococcus aureus
Colony Characteristics HEMOLYSIS ON BAP

Streptococcus pyogenes Streptococcus agalactiae

Most important in presumptive identification of Streptococci and

enterococci
Growth of Organisms in Liquid Media

A, Clear broth indicating no

bacterial growth (left) and turbid
broth indicating bacterial growth
(right).

• TURBIDITY Turbidity produced by

enterics when growing
– Overall cloudiness of liquid in thioglycollate.
Growth of Organisms in Liquid Media

“Vine” or “streamer” “Puffed balls”

Growth of Organisms in Liquid Media

“scum”

Yeast Pseudomonas spp.

Growth of Organisms in Liquid Media

Yeast
Examples of Distinctive
Colony Characteristics
Distinctive Colony Characteristics
Swarming: hazy blanket
of growth on surface of
Proteus spp.
Distinctive Colony Characteristics

Candida albicans
Distinctive Colony Characteristics

“greenish
metallic
sheen”

Escherichia coli in EMB

Distinctive Colony Characteristics

Encapsulated Klebsiella pneumoniae

“mucoid colonies”
Distinctive Colony Characteristics
the end