CULTURE MEDIA

Learning Objectives

 Classify culture media.

 Discuss the types of solid culture media
with their examples.
 Listthe steps for the preparation of
Culture media.
 Discuss the method of Culture
inoculation (Streaking).
 These are special preparations used to
grow microorganisms in the laboratory

CLASSIFICATION
 Solid Media
 Semisolid Media
 Liquid (Broth) Media
 Culture media are solidified by
incorporating a gelling agent such as
 Agar
 Gelatin
 SOLID MEDIA

Solid media contain agar concentration

of 1.5% w/v
Uses:
 Isolate discrete colonies of bacteria
present in the specimen
 Determine the colony characteristics of
the isolate
 SEMISOLID MEDIA

They have soft consistency and contain

agar concentrations of 0.4-0.5%
Uses:
 Used mainly as transport media
 For motility & biochemical tests
 LIQUID (BROTH) MEDIA

These media do not contain gelatin or

agar.
Uses:
 commonly used as enrichment where
organisms are likely to be few e.g. blood
culture
 Biochemical testing e.g. fermentation
reactions
 TYPES OF SOLID MEDIA

 Basic media

 Enriched media

 Selective media

 Differential (Indicator) media

 BASIC MEDIA

Simple media that support the growth of

microorganisms, which do not have
special nutrient requirements.

Examples: Nutrient agar

 NUTRIENT AGAR
TYPE:
Basic culture medium
COMPOSITION:
• Beef extract
• Peptone
• NaCl
• Agar
USES:
 Antibiotic sensitivity
testing

 Subculture of bacteria
 ENRICHED MEDIA

Contain whole blood, lysed blood, serum

or special extracts to support the growth
of bacteria which require additional
nutrients (fastidious organisms)
Examples:
 Blood agar
 Chocolate agar
 BLOOD AGAR
TYPE:
Enriched & Differential
medium
COMPOSITION:
 Beef extract
 Peptone
 NaCl
 Agar
 Defibrinated hemolysis free
blood
USES:
 to grow fastidious
bacteria e.g.
streptococci
 to see hemolysis of
bacteria
BETA HEMOLYSIS ON
BLOOD AGAR
 CHOCOLATE AGAR
TYPE:
Enriched culture medium
COMPOSITION:
 Beef extract
 Peptone
 NaCl
 Agar
 Defibrinated hemolysis
free blood
USES:
 When the blood agar is heated, RBCs
are broken down and factors V and X
are released.
 Required for the growth of some
bacteria e.g. Neisseria, Haemophilus and
Pneumococci
 ENRICHMENT MEDIA

Liquid media that increase the

number of specific pathogens due
to the presence of certain
substances that discourage the
multiplication of unwanted
bacteria
 TRYPTONE SOYA BROTH

EXAMPLES:
 Tryptone soya broth and
Brain heart infusion are
used as enrichment
medium for blood culture

TRYPTIC SOYA BROTH

 SELECTIVE MEDIA

These media favour the growth of particular

bacteria and inhibit the growth of other
bacteria
EXAMPLES:
 MacConkey agar
 Lowenstein-Jensen medium
 TCBS medium(thiosulfate-citrate-bile-salt
sucrose agar)
 DIFFERENTIAL (INDICATOR)
MEDIA
These are the media to which indicators, dyes or
other substances are added to differentiate
between bacteria

EXAMPLES:
 MacConkey agar
 CLED (cystine-lactose-electrolyte-deficient
agar)
 TCBS
 MACCONKEY AGAR
TYPE:
Selective & differential
culture medium
COMPOSITION:
 Beef extract
 Peptone
 NaCl
 Lactose
 Crystal violet
 Bile salts
 Crystal violet
 Neutral red (indicator)
 Agar
SELECTIVE: allows the growth of gram
negative bacteria & inhibits the growth of
gram positive bacteria
Crystal violet & bile salts inhibit the growth of
gram positive bacteria
DIFFERENTIAL: on the basis of lactose
fermentation
 Lactose fermenters produce pink colour
colonies
 Non-Lactose fermenters produce colorless
colonies
LACTOSE AND NON-LACTOSE
FERMENTERS ON MACCONKEY AGAR
CYSTEINE LACTOSE ELECTROLYTE
DEFICIENT (CLED) AGAR
TYPE:
Differential culture medium

COMPOSITION:
 Beef extract
 Peptone
 Lactose
 Cysteine
 Bromothymol blue & Andrade
(indicator)
 Agar
DIFFERENTIAL: on the basis of lactose
fermentation
 Lactose fermenters produce yellow colour
colonies
 Non-Lactose fermenters produce whitish blue
colonies
USED SPECIFICALLY FOR URINE
CULTURE
 For the isolation and enumeration of
urinary pathogens
 Itis made electrolyte deficient i.e. does
not contain NaCl
 Inhibits swarming motility of Proteus so
that colony count can be done
LACTOSE AND NON-LACTOSE
FERMENTERS ON CLED
 LOWENSTEIN-JENSEN
MEDIUM
TYPE:
Selective culture medium for
Mycobacterium tuberculosis
COMPOSITION:
 Egg white
 L-Asparagine
 Potato starch
 Glycerol
 Malachite green
 Penicillin
 Always in screw cap bottle with slant
 Glycerol promotes the growth of
mycobacterium tuberculosis
 Malachite green and penicillin kill
unwanted bacteria
 Mycobacterium tuberculosis produces
dry, rough and buff colonies in 6-8
weeks.
THIOSULPHATE CITRATE
BILE SALT SUCROSE
(TCBS) AGAR

Selective & Differential
medium for Vibrio cholera
 Yellow colored colonies
of vibrio cholera (Sucrose
fermenter)
 Other Vibrio species give
green colored colonies
 Alkaline medium (pH
>8.4)
SABOURAUD DEXTROSE AGAR

 Selective medium
used for isolation
of various fungi.
 Alkaline pH
 TRANSPORT MEDIA

 Mostly semi-solid media

 Contain ingredients to prevent overgrowth &
ensure the survival of pathogens when
specimens cannot be cultured soon after
collection
EXAMPLES:
 Cary-Blair medium
 Amies medium
 PREPARATION OF CULTURE
MEDIA

 Ready-made standardized
dehydrated media are
used.
 Write on label date of
receipt in laboratory.
 Store below 25 ◦C in a dry
area away from sunlight.
 Check expiry date on the
label.
 STREAKING
 Culture inoculation technique
 All culture media must be checked visually
before use for contamination.
 Use of an aseptic technique
 Make sure plate is free of droplets
 WIRE LOOP

USES:
 Smear Preparation
 Culture Inoculation
 PRIMARY
STREAK

SECONDARY
STREAK
WELL

TERTIARY
STREAK