Mueller-hinton Broth: Howto

Mueller Hinton broth is used as the liquid medium for reviving the microorganism for antibiotics sensitivity testing, for determination dilution studies. It is also developed for the cultivation of nesserias and other fastidious microorganism due to the richness of the medium.

Formula (g/Liter):

Equipment:
Conical flask
Heater/stirrer
Glass rod
Autoclave

Preparation:

1. Dissolve the ingredient mix in one liter of distilled water while stirring constantly.
2. Heat them with frequent agitation and boil for one minute
3. Autoclave at 121C (15 lbs.sp for 15 minutes), DO NOT OVERHEAT.
4. Cool down, and measure the pH. It should be 7.4 at 25C.

Validation test:
A good mueller-hinton broth will show growth of any of the following microbes:

• Staphylococcus aureus ATCC25923,
• Escherichia coli ATCC 25922,
• Streptococcus faecalis ATCC 33186,
• aeruginosa ATCC27835,
• Streptococcus pyogenes ATCC 19615,
• Listeria monocytogenes ATCC19113.