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