Bacteriological Culture Media

Objectives of bacterial culture

1. To study bacterial morphology, physiology and pathogenic properties.
2. At the clinical level
 Isolation of bacteria in pure culture
 Identification
 Antimicrobial susceptibility
3. Preparation of important bacterial products, e.g., toxins, antigens,
vaccines …

 According to the physical state media may be:

1. Liquid media
2. Solid media

 Solid media are preferable because:

1. They help identification of the organism (different colonial
morphology).
2. They allow isolation of the organism in pure culture for further studies.
BATERIOLOGICAL LOOP

CULTIVATION OF BACTERIA ON SOLID MEDIA

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Types of culture media:
1. Simple media
2. Enriched media
3. Selective media
4. Enrichment media
5. Selective and indicator media

1- SIMPLE MEDIA
1. Nutrient broth
2. Nutrient agar
3. Peptone water

1- NUTRIENT BROTH

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 2- NUTRIENT AGAR

2- ENRICHED MEDIA
 Significance: enriched media help growth of fastidious organisms
e.g. Streptococci
1. Blood agar
2. Chocolate agar

1- Blood agar
Enriched substance: blood

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 1. Complete haemolysis (Beta haemolysis)

 Complete haemolysis of RBCs (clear zone around colonies)

 Example: Strept. pyogenes
2. Alpha hemolysis

 Partial haemolysis due to conversion of hemoglobin to billiverdin

(greenish discoloration)
 Exmples:
1. Strept. Pneumoniae
2. Viridans streptococci
3. Non hemolytic organism

 Example: enterococci

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 2- Chocolate agar
 Enriched substance: heated blood

3- SELECTIVE MEDIA
 Significance:
Selective media contain selective substances which inhibit the growth of
some bacteria but have no effect on the organism of interest , allowing it to
grow.
1. Lowenstein-Jensen medium
2. Blood tellurite
3. Modified Thayer-Martin

1- Lowenstein Jensen medium

 for Mycobacterium tuberculosis (M. tuberculosis)
 Selective substance: malachite green dye

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 2- Blood tellurite
 Blood agar made selective by K tellurite
 For Corynebacterium diphtheria

3- Modified Thayer-Martin medium

 Chocolate agar made selective by antibiotics
 For neisseria

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 4- ENRICHMENT MEDIA
 Liquid media with selective property
 Selenite F Broth: to help growth of salmonella & shigella in stools
samples

5- SELECTIVE AND INDICATOR Media

 They have both
1. Selective
2. indicator property:indicator media contain substances that can
differentiate between two or more types of bacteria.
 MacConkey’s medium
 Mannitol salt agar
 TCBS

1- MacConkey’s medium
 Selective substance: bile salts
 Test sugar: lactose
 pH indicator: neutral red

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 Lactose fermenter on MacConkey’s medium
 Example: Escherichia coli, Klebsiella

Lactose non fermenter on MacConkey’s medium

 Example: Salmonella, shigella, proteus

MacConkey’s medium

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 2- Mannitol salt agar
 Selective substance: salt (NaCl) for staphylpcocci
 Test sugar: mannitol
 pH indicator: phenol red

3- Thiosulfate Citrate Bile salt Sucrose agar

(TCBS)
 Selective substance: Thiosulfate, Citrate and bile salts + alkaline pH
(for vibrios).
 Test sugar: sucrose
 pH indicator: bromothymol blue

Sucrose fermenter on TCBS (Vibrio cholera)

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 Sucrose non-fermenter on TCBS (other vibrios)

ANAEROBIC CULTIVATION
 ANAEROBIC MEDIA Robertson cooked meat
 Examples:
1. Bacteroids
2. Clostridium tetani

GAS PAK SYSTEM FOR ANAEROBIC CULTIVATION

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