B.H.I.A. (BRAIN HEART INFUSION AGAR)
W/ CHLORAMPHENICOL AND CYCLOHEXIMIDE

Technical Data #1057a / 98.04.22

B.H.I.A. with chloramphenicol and cycloheximide is a selective solid medium for isolation of pathogenic fungi from clinical specimens contaminated with bacteria and non-pathogenic molds. Chloramphenicol and cycloheximide inhibit gram-positive, gram-negative bacteria and some yeasts.

FORMULA
in grams per liter purified filtered water

Brain Heart Infusion	10,0
Casein Peptone	8,5
Meat Peptone	8,5
Sodium Chloride	5,0
Dextrose	2,5
Disodium Phosphate	2,5
Agar	15,0
Antibiotics
Chloramphenicol	0,05
Cycloheximide	0,5

pH 7.4 +/- 0,2 à 25º C

This approximate formula may be adjusted and/or enriched to obtain best results.

PRECAUTIONS
This medium is for IN VITRO diagnostic use only.

STORAGE
Store prepared media at 2-8º C protected from direct light and dehydrated powder, in a dry place, in tightly-sealed containers at 2-25º C.

SIGN OF DETERIORATION
Media should not be used if the expiry date has passed. Prepared media should not be used if there are signs of contamination or deterioration (shrinking, cracking, evaporation or discoloration). Do not use dehydrated media if it is caked.

DIRECTIONS
Suspend 52 g of dehydrated media in 1000 ml of purified filtered water. Heat with frequent agitation and boil for one minute. Sterilize at 121º C for 15 minutes. Cool to 45-50º C and aseptically, add 30 ml of chloramphenicol (8629) and 25 ml of cycloheximide (8724). Mix gently and dispense into sterile Petri dishes or sterile tubes.

PROCEDURE
Prior to inoculate, the prepared media should be brought to room temperature.

1. Directly, inoculate plates as soon as possible with specimen and streak for isolation. Cutaneous specimens should be pressed lightly into medium.
2. Seal plates with masking tape, incubate at 25º C for 4-6 weeks. Incubate bottles with loosened caps.
3. Examine weekly for growth.

QUALITY CONTROL
Results after 7 days at 25ºC

Organisms	ATCC	Growth
Candida albicans	10213	+
Candida albicans	60193	+
Trichophyton mentagrophytes	9533	+
Candida krusei	14243	-
Escherichia coli	25922	- or partial

LIMITATIONS OF METHOD
This medium is only a part of the identification. Other tests may be required.

REFERENCES

1. Ajello, J., L.K. Georg, W. Kaplan, L. Kaufman. 1963 CDC Laboratory manual for medical mycology. PHS Publication No. 994. US Govt. Printing Office, Washington. D.C.
2. Finegold, S.M., E.J. Baron 1986 Bailey and Scott's Diagnostic Microbiology 7th ed. C.V. Mosby, St. Louis
3. Lennette, E.H., Ballows, A., Hausler, W.J.Jr., and Shadomy, H.J. Manual of Clinical Microbiology. 4th ed. 1985 Washington D.C.: American Society for Microbiology.
4. Mac Faddin, Jean. F., 1985 Media for Isolation-Cultivation-Identification-Maintenance of Medical Bacteria. Vol. 1. Baltimore, MD.: Williams & Wilkins.

CATALOGUE NUMBERS

Dehydrated media	500 g	QB-39-0206
Prepared media
Plates, 100x15 mm	10/pkg	1057
Mono-Plates, 100x15 mm	10/pkg	1057M
Tubes, 20x125 mm	10/pkg	2489
Antibiotics
chloramphenicol, 10 ml	10/pkg	8629

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