Culture Media

Dr. Qurat-ul-Ain Ayaz

 DEFINITION
These are the preperations which provide food
and nutrition to bacteria when they are grown in
the lab
 Classifiation
Based on Consistency
Based on Utility/ Composition
CLASSIFICATION BASED ON CONSISTENCY

 Liquid media. These are fluid in nature, e.g.

Nutrient broth and alkaline peptone water
 Solid media. Prepared by adding solidifying
agents like agar and gelatin to the liquid
medium e.g. Nutrient agar. Agar is an inert
polysachharide obtained from seaweeds. It is
used to solidify culture media. It has high
melting temperature, which enables petri
dishes to be kept in the incubator
 Semi-solid media. This form of culture
medium is prepared by adding a small amount
of agar (0.4-0.5% w/v) to a fluid medium.
Semi solid media are mainly used as transport
media and for motility and biochemical tests
CLASSIFICATION OF CULTURE MEDIA BASED
ON UTILITY/COMPOSITION
• Simple (Basic) Media:
– It is used to grow organisms that do not have special
nutritional requirements. e.g nutrient agar, nutrient
broth and peptone water.
– Nutrient agar (Solid Media):
• It is used to grow micro-organisms that have no special
nutritional requirement (non-fastidious bacteria). It is
also used for sensitivity testing of drugs.
• Composition: Beef extract, peptone water, NaCl, Agar.
• Micro-organism cultured: Staphylococcus spp.,
Pseudomonas spp.
 CLASSIFICATION OF CULTURE MEDIA BASED
ON UTILITY/COMPOSITION
• Enriched Media:
– These are the media that are enriched with substances like
whole blood, lysed blood serum or special extracts to
support growth of organisms. e.g blood agar, chocolate agar.
– Blood Agar (Solid Media):
• It is used to grow those organisms that are more difficult to grow
on ordinary media such as Streptococcus pneumonia. Blood agar is
also used to study the hemolytic property of bacteria.
• Composition: Melted agar, sterile defibrinated blood 5-10%, Beef
extract, Peptone water, NaCl.
• Micro-organisms Cultured: Streptococcus pyogenes,
Streptococcus viridians, and Streptococcus pneumonia
BLOOD AGAR
CLASSIFICATION OF CULTURE MEDIA BASED
ON UTILITY/COMPOSITION
– Chocolate Agar (Solid Media):
• When blood agar is heated, the red cells are lysed and
the medium becomes brown in color. It is referred to as
chocolate agar
• Composition: Same as blood agar.
• Micro-organisms Cultured: X and V factors are
released by lysis of red blood cells that are essential for
growth of some bacteria like H.influenzae. Neisseria
gonorrhoae, Neisseria meningitides require lysed blood
agar for their growth.
 CLASSIFICATION OF CULTURE MEDIA BASED
ON UTILITY/COMPOSITION
• Differential (Indicator) Media:
– It is used to differentiate between two groups of
bacteria. e.g MacConkey’s Agar and CLED
– MacConkey’s Agar (Solid Media):
• It is a differential as well as selective medium, used to
distinguish lactose, non-lactose and late lactose
fermenters.
• Composition: Nutrient agar, Bile salts, Lactose, NaCl, Neutral
red (as indicator), Peptone
• Micro-organism Cultured: Enterobacteriaceae group,
Klebsiella pneumoniae, Pseudomonas, Yersinia species, and
Enterococci.
CLASSIFICATION OF CULTURE MEDIA BASED
ON UTILITY/COMPOSITION
– CLED (Solid Media):
• This is short form for Cystine Lactose Electrolyte
Deficient media. It is used to culture bacteria from
infected urine. It differentiates between lactose
fermenters (yellow) and non-lactose fermenters (blue,
grey, or green). It also inhibits swarming of proteus.
• Composition: L-cystine, Bromothymol blue (indicator),
lactose
• Micro-organisms cultured: E. coli, Staphylococcus,
Streptococcus, and Candida
 CLASSIFICATION OF CULTURE MEDIA BASED
ON UTILITY/COMPOSITION
• Selective Media:
– Contain substances such as bile salts or antibiotics that inhibit the
growth of some organisms but have little or no effect on the
required organism. Used for clinical specimens such as faeces.
– Solid Media:
• Thiosulphate citrate bile salt sucrose (TCBS) Agar:
– It is a selective medium used to isolate Vibrio cholera and other Vibrio species.
– Composition: Yeast extract powder, Bacteriological peptone, Sodium thiosulphate,
Sodium citrate, Ox-bile, Sucrose, NaCl, Ferric Citrate, Bromothymol blue, Agar
– Color: Light green
– Micro-organisms cultured: Vibrio cholera produces yellow colonies on TCBS.
• Sabouraud’s Dextrose Agar :
– This medium is used to grow fungus. The pH is kept acidic.
– Composition: Peptone ….10g, Dextrose…..40g, Agar…..15g.
– Color: Pale yellow
CLASSIFICATION OF CULTURE MEDIA BASED
ON UTILITY/COMPOSITION

– Semi Solid Media:

• Lowenstein Jensen (L J) Medium:
– L J medium is used to isolate Mycobacterium tuberculosis.
– Composition: Whole egg, Penicillin (inhibit growth of other bacteria),
Malachite green (it is also antibacterial at low pH), Glycerol (to enhance
growth of human type of M. tuberculosis), K2Mg4, MgSO4, Mg citrate, potato
flour, asparagine
– Color: Light green (always in screwed bottle with slant)
– Micro-organism cultured: Mycobacterium tuberculosis takes 14-21 days to
produce dry, buff, raised and waxy colonies.
– Liquid Media:
• Alkaline Peptone Water:
– It is used as a selective media for Vibrio cholera.
– Composition: Peptone….5g, Sodium chloride…..5g, distilled water….500ml.
 CLASSIFICATION OF CULTURE MEDIA BASED
ON UTILITY/COMPOSITION

• Enrichment Media:
– Broth media with selective properties, enriching the desired
range of organisms and inhibiting others. Examples include TT
Broth and Selenite F Broth.
• Cold enrichment: Listeria monocytogenes
• Heat enrichment: Legionella species
– Robertson’s Cooked Meat Medium (Liquid Media):
• Composition: Minced meat, Broth, Paraffin oil, NaOH (to neutralize
lactic acid)
• Color: Pale yellow, with minced meat at bottom
• Micro-organisms cultured: Clostridium species, Bacteroides, other
anaerobes and facultative anaerobes.
• Transport Media:
– These are mostly semisolid media that contain
ingredients to prevent the overgrowth of
commensals and ensure the survival of aerobic
and anaerobic pathogens when specimens cannot
be cultured immediately after collection.