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Mitis-Salivarius Agar
Purpose:

Mitis Salivarius Agar, when used with Chapman Tellurite Solution (1%), is a selective
medium for the isolation of Streptococcus oralis, Streptococcus salivarius and
enterococci from mixed cultures. It is used for differentiation of the viridans strains and
also for epidemiological microbial studies on dental plaque and carious lesions from
saliva specimens and for etiological agents of dental caries.

Principle:

This medium contains the selective agents crystal violet, potassium tellurite and trypan
blue. These agents inhibit most gram-negative bacilli and most gram-positive bacteria
except streptococci. Sucrose is incorportated in a 5% concentration which is utilized as
an energy source and enables Streptococcus salivarius to develop more luxurient
colonies than Streptococcus oralis or enterococci.

Procedure:

Inoculate the sample onto a room temperature Mitis Salivarius Agar plate and streak for
isolation in the usual manner. Incubate anaerobically in a GasPak jar (for Streptococcus
salivarius and S. mutans recovery) or in a candle jar for 48 hours at 35-37C.

Interpretation:

Streptococcus salivarius - Large, pale blue, opaque, mucoid colonies (classical "gum-
drop" appearance due to formation of levan from sucrose).

Streptococcus oralis - Blue, small, flat, hard colonies with a dome center.

Streptococcus mutans - Raised, convex, undulate, opaque, pale blue colonies, granular
"frosted glass" appearance. May exhibit a glistening bubble on surface of colony due to
excessive synthesis of glucan from sucrose.

Streptococcus sanguis - Raised, smooth, hard colonies embedded in agar.

References:

REMEL Microbiology Products. 1990. Technical Manual, Technical Information (TI

No. 1615-A). Lenexa, KS.

MacFadden, Jean F. 1985. Media for isolation-cultivation-identification-maintenance of

medical bacteria, Vol. 1, Williams and Wilkins, Baltimore, MD.
 S. salivarius on M-S agar: note colony morphology and texture

Glistening "gum-drop" colonies of S. salivarius on M-S agar.

S. oralis on M-S agar: note colony morphology and texture.

S. mutans on M-S agar: note colony morphology and texture.

Strep.mutans: dextran pools

Strep. mutans: typical fresh isolates.

S. sanguis on M-S agar: note colony morphology and texture.