Mueller Hinton Agar

In 1941, Mueller and Hinton developed a protein free medium for isolating

pathogenic strains called Mueller Hinton Agar. The medium was useful for identifying

sulfonamide-resistant and responsive strains from gonococci (Ryan, 1970). Moreover

Bauer, Kirby, et al. (1960) described Mueller Hinton Agar as commonly used for

standardized in antimicrobial disk susceptibility testing. Barry and Fay (1973)

investigated the effects of changing the depth of plated Mueller Hinton Agar on disk

diffusion testing. Dewees, et al(1970) studied the effect of cache on Mueller Hinton Agar

plates that are used for antimicrobial disk diffusion zone sizes. Their findings were

suitable for usage in routine susceptibility testing.

Mueller Hinton Agar plates are also used alongside with Kirby-Bauer

antimicrobial disk diffusion procedure. It is also based on usage of an antimicrobial

impregnated filter paper disk. The effectiveness of the antimicrobial can be shown by

measuring the zone of inhibition of pure culture of an organism. When an impregnated

disk is placed on agar surface, it can result to the diffusion of the antimicrobial into the

surrounding medium. Zone diameters established for each antimicrobial determining

resistant, intermediate, and sensitive results for pathogenic microorganisms.

Mueller Hinton Media contains beef infusion, starch, and casamino acids.

The beef infusion and casamino acids serve energy and nutrients while the starch acts

as a colloid which protects against toxic material in the medium. The agar is only added

when solidifying agent is needed. The levels of tetracycline and sulfonamide inhibitors,
thymidine, thymine, magnesium, and calcium ions, are controlled for it is not to interfere

with susceptibility testing and to yield good growth (Haltiner, et al., 1980).

Mueller Hinton Broth is the same formulation, just without the added agar.

It is used for the cultivation of microorganisms, and for making liquid out from organisms

to be used in the Kirby-Bauer disk diffusion procedure.

The stores plates are recommendable to be stored at 2-8ºC away from

direct light. The tube media may be stored at 2-30ºC. The media should not be used if

there are shrinking, cracking, or discoloration), contamination, or if the expiration date

has passed. The product is light and temperature sensitive, so it must be protected from

light, excessive heat, moisture, and freezing.

It is recommended that biochemical, immunological, molecular, or mass

spectrometry testing will be performed on colonies that pure cultured for complete

identification. The disk diffusion method also should not be used for obligatory

anaerobes, slow growing organisms, and capnophiles. This method is standardized for

facultative organisms or rapid growing aerobes.

Barry and Fay. 1973. Am. J. Clin. Pathol.

Bauer, A.W., W.M.M. Kirby, et al. 1966. Am. J. Clin. Pathol.

Dewees, et al. 1970. Effect of storage of Mueller Hinton Agar plates on zone sizes for

antimicrobial testing. Appl. Microbio

Haltiner, R.C., P.C. Migneault, and R.G. Roberston. 1980. Incidence of Thymidine-

Dependent Enterococci Detected on Mueller-Hinton Agar with Low Thymidine

Content. Anti. Ag. and Chemo.

Mueller, J.H. and J. Hinton. 1941. A protein-free medium for primary isolation of

the Gonococcus and Meningococcus Proc. Soc. Exp. Diol. and Med

 Ryan, K.J., et al. 1970. Disk sensitivity testing. Hosp. Prac.