MD-2021 | Manila Central University S.Y.

2018-2019

MICROBIOLOGY COMPOSITION
CULTURE MEDIA CLASSIFICATION DESCRIPTION
Dr. Catherine Gironella | September 5, 2018
Natural Media Use ingredients from natural sources
such as meat, fruit, milk, etc.
CULTURE MEDIA Example:
- Isolation and identification of pathogens with - Potato infusion broth
biochemical tests are still considered the gold Synthetic or Defined Use chemical constituents which are
standard in the diagnosis of diseases Media qualitatively and quantitatively known
Examples:
- E.coli complex medium
MEDIA CLASSIFICATION
CONTAINER
According to:
CLASSIFICATION DESCRIPTION
- Physical State Plated Media Solid media placed in a petri dish
- Application or Function Tubed Media Solid or liquid media placed in a tube
- Composition Media could be a slant/butt, butt or
- Container broth

PHYSICAL STATE
CLASSIFICATION DESCRIPTION COMMONLY USED MEDIA FOR ISOLATION OF
Liquid Without solidifying agent BACTERIAL PATHOGENS
Semi-solid Contains about 0.5-1.0% agar
Solid Contains about 1.5-3.0% agar EMB (EOSIN METHYLENE BLUE)
- Selective and differential medium
- Selective:
APPLICATION or FUNCTION o Aniline dyes (eosin and methylene blue)
CLASSIFICATION DESCRIPTION which inhibiti gram-positive and fastidious
Ordinary Nutrient Many common organisms can grow gram-negative organisms
Media (Simple) Example: Nutrient agar or broth - Differential:
Enriched Media It duplicates the natural environment of o Contain lactose
organism o Lactose-fermenting organisms have pinkish
Additional nutrients (blood, serum or colonies
albumin) are added to the ordinary ▪ E. coli produce green black with
medium
metallic sheen
Media with a chemical composition
▪ Metallic sheen due to continuous
inhibit or kill the normal intestinal flora
like coliforms fermentation of lactose
Examples:
- CAP, BAP
- LJ medium

*a chemical composition inhibit or kill

the normal intestinal flora like the
coliforms and permit Salmonella and
some Shigella to grow unrestricted
o Non-lactose fermenting organisms have the
Example: Selenite Broth
transparent or colorless colonies
Selective Media Complex media to differentiate among
certain genera but are highly selective
for the organism MacCONKEY AGAR
Examples: - Enriched:
- SSA, DCA, TCBS o Contains bile salts and crystals which inhibit
Differential Media Contains chemicals that characterize gram-positive and fastidious gram-negative
the bacteria by their distinctive colonial bacteria
appearance in culture - Differential:
Examples: o Lactose-fermenting organisms appear pnk
- EMB and non-lactose fermenters are colorless or
- MacConkey for lactose transparent
fermentation (pinkish colonies
for lactose fermenters)

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 MD-2021 | Manila Central University S.Y. 2018-2019

- Klebsiella appear pinkish mucoid colonies o Salmonella colonies are colorless with black
precipitate due to lactose fermentation and
formation of H2S

DCA (DEOXYCHOLATE CITRATE AGAR)

- Highly selective for Salmonella
o High concentration of bile salt and sodium
deoxycholate
o Sodium and ferric citrate prevents the growth
of E.coli
- Differential for H2S-producing organisms
ENDO AGAR
TCBS (THIOSULFATE CITRATE BILE SALTS SUCROSE
- Enriched:
AGAR)
o Contains sodium sulfite and basic fuchsin
(makes the colonies appear reddish pink)
which inhibits the growth of Gram-positive
bacteria
o Klebsila will appear reddish instead of
pinkish as in Mac
o E.coli appear dark red at first before
producing the classic green metallic sheen
o Salmonella is colorless (NLF)
- Differential:
o Lactose-fermenting colonies appear pink to - Highly selective and enriched medium for Vibrio
rose red o Citrate and bile salts inhibit Gram-positive
o Strong acid producers may produce a and most of the Gram-negative bacteria
metallic sheen from a reaction with basic - Differential for those species which ferment sucrose
fuchsin o Sucrose-fermenting organisms appear
o Non-lactose fermenters appear colorless to yellow
faint pink o Non-sucrose fermenting organisms appear
bluish green (same as medium)
SSA (SALMONELLA SHIGELLA AGAR) o V. cholera vs V. parahemolyticus
o V. parahemolyticus does not ferment
sucrose

MSA (MANNITOL SALT AGAR)

- Highly selective and enriched

o High bile salts and sodium citrate inhibit all
Gram-positive bacteria and many Gram-
negative organisms including the coliforms
- Differential
o Sodium thiosulfate is the source of sulfur.
o Bacteria that produce H2S form black
precipitate (FeS) with ferric citrate - Highly selective for Staphylococcus
o Shigella produces colorless colonies without o 7.5% salt concentration will inhibit most
black precipitate bacteria
o S. aureus is halophilic (salt-loving)

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 MD-2021 | Manila Central University S.Y. 2018-2019

- Differential for those species which ferment mannitol - Neisseria and Haemophilus grow best in this medium
o S. aureus and S. saprophyticus are
mannitol-fermenters → appear as yellow
colonies

LJ (LOWENSTEIN-JENSEN MEDIUM)
- Selective and enriched for Mycobacteria
- Enriched
o Contains egg; the malachite green inhibits
the growth of most of contaminants
o Crust-like

BAP (BLOOD AGAR PLATE)

- Enriched medium
o Allows the growth of both non-fastidious and
some e fastidious organisms
- Differential for those organisms which produce alpha- CHOPPED MEAT MEDIUM
hemolysis or beta-hemolysis

o Incomplete hemolysis – α-hemolysis (green)

o Complete hemolysis (clear zone near
colonies) – β-hemolysis
o No hemolysis – γ-hemolysis
Proteus – characteristic spread in BAP
- Used to enrich and preserve the growth of anaerobic
organism
- Meat particles – substrate for proteolytic enzymes
which serve as reducing substances to maintain low
Eh (oxidation-reduction potential) and prevent rapidly
growing bacteria from overgrowing slower forms
- Put mineral oil on top to prevent oxygen from entering
the medium

BHI (BRAIN HEART INFUSION MEDIUM)

- Nutritionally rich formula for wide variety of
microorganisms
- Broth or solid
- Can be used as base for BAP
- Agar may also be used for primary isolation of fungi
- For transport

CAP (CHOCOLATE AGAR PLATE)

- Contains RBCs which are partially lyzed by heating at
85oC liberating hemoglobin, hemin, and coenzyme
nicotine adenine dinucleotide

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 MD-2021 | Manila Central University S.Y. 2018-2019

MHA (MUELLER-HINTON AGAR) - Inside the gaspak:

- Used in antibiotic susceptibility testing o O2 trapped acts on H2 to form H2O (seen as
water droplets on the side of the container)
▪ Process continues until the strip
becomes white
▪ If it’s already white, the gaspak may
be incubated
- Candle jar

o Uses a lighted candle to use the oxygen in

ANAEROBIC CULTURE the container
- Used in isolation of organisms which cannot tolerate ▪ Combustion needs O2 in order to
O2 proceed
- Media that can be used: ▪ The candle turns off when there is
o Chopped meat medium – most common no more O2
o Blood agar ▪ Container is sealed before
- Anaerobic gaspak or Gas pouch incubation

- Catalyst and indicator slip

o Speed up the reaction to produce gases
such as nitrogen, H2 and CO2
o Palladium – catalyst
- Indicator slip
o Turn to blue in the presence of oxygen

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