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SELECTIVE, DIFFERENTIAL

AND ENRICHED MEDIA


• Selective media: used to select (isolate) specific
groups of bacteria;
• chemical substances in the media inhibit the growth
of one type of bacteria while permitting growth of
another (MSA, EMB, MacConkey)

• Differential media: distinguishes among


morphologically and biochemically related groups of
organisms;
• chemical compounds (following inoculation and
incubation) produce a characteristic change in the
appearance of bacterial growth and/or the medium
surrounding the colonies (MSA, EMB, MacConkey)

• Enriched media: supplemented with highly nutritious


materials, such as blood, serum, or yeast extracts, for
the cultivation of fastidious organisms
Mannitol Salt Agar
(MSA)
• Differentiates
• Mannitol salt agar is both Staphylococcus species,
selective and differential by mannitol fermentation

• Selective: It favors • (S. aureus ferments,


organisms capable of S. epidermidis does not)
tolerating high salt
concentrations • Phenol red is the pH
(7.5 % NaCl) indicator

• Differential: It Neutral - Basic pH


distinguishes bacteria red at 7.4 to 8.4
based on their ability to
ferment mannitol
Acidic pH
yellow below 6.8
RESULTS:
• Positive Results:
The development of “yellow halos” around the
bacterial growth means mannitol has been
fermented and acid end products have been
produced (S. aureus)

• Negative Results:
No color change in the medium is a negative
result (S. epidermidis)

• No growth on the medium indicates a Gram-


organism (E. coli)
PROCEDURE:
On an MSA Plate:
S. aureus (growth, yellow)

E.coli S.epidermidis
(No growth) (growth, red)
Eosin Methylene Blue Agar (EMB)
• Eosin methylene blue agar is a Typically used for the family
selective and differential Enterobacteriaceae—
medium enteric (gut) bacteria (Gram-
• Selective: EMB contains rods)
the dyes methylene blue
and eosin which inhibit They may be divided into
Gram + bacteria, thus those that produce acid from
favoring growth of Gram – lactose fermentation
(coliforms) and those that do
• Differential: EMB contains not
lactose, thus allowing for
the distinction between Coliforms include
lactose fermenters and Escherichia coli and
nonferments
Enterobacter aerogenes
Results
• Large amounts of acid from lactose
fermentation cause the dyes to
precipitate on the colony surface,
producing a black center or a “green
metallic sheen” (E. coli)

• Smaller amounts of acid production


result in pink coloration of the
growth (E. aerogenes)

• Nonfermenting enterics do not


produce acid so their colonies
remain colorless or take on the color
of the media (P. vulgaris)

• No growth indicates a Gram +


organism (S.aureus)
•Large amounts of acid from lactose fermentation cause
the dyes to precipitate on the colony surface, producing
a black center or a “green metallic sheen” (E. coli)
PROCEDURE:
On an EMB Plate:

P. vulgaris S. aureus

(growth, (no growth)

no color)

E.coli
E. aerogenes
black colonies or green black colonies or
metallic sheen green metallic sheen
MacConkey Agar
(MAC)
• A selective and
differential medium • Bile salts and crystal
violet inhibit growth of
used to isolate G+ organisms
members of the (selective)
Enterobacteriaceae
• Neutral red is a pH
indicator that is
• Contains nutrients, colorless, but yellow
including lactose, as above pH 8 and red at
well as bile salts, pH less than 6.8
neutral red and (differential)
crystal violet
• Acid accumulating from lactose fermentation
turns the colorless neutral red to a red color—
therefore coliforms produce a red “halo” on the
medium (E.coli, E.aerogenes)

• Lactose nonfermenters will grow, but don’t


produce acid. Therefore, the neutral red remains
colorless (P. vulgaris)

• No growth indicates a
Gram + organism
(S.aureus)

Macconkey agar with lactose(left) and non-lactose(right) fermenters


PROCEDURE:
• On a MacConkey Agar Plate:
P.vulgaris (colorless) S.aureus (no growth)

E.aerogenes E.coli (Red)


(Red)