B.H.I.A. (BRAIN HEART INFUSION AGAR)
W/ 10% SHEEP BLOOD CHLORAMPHENICOL,
CYCLOHEXIMIDE AND GENTAMYCIN

Technical Data #1031a / 98.04.22

B.H.I.A. w/ 10% sheep blood, chloramphenicol, cycloheximide and gentamycin is a selective solid medium for isolation of pathogenic fungi from clinical specimens contaminated with bacteria and non-pathogenic molds. Chloramphenicol, cycloheximide and gentamycin 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

Defibrinated Sheep Blood

100 ml

Antibiotics

 

Chloramphenicol

0,016

Gentamycin

0,005

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 900 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 100 ml of Defibrinated Sheep Blood (4147), 10 ml of chloramphenicol (8629), 10 ml of gentamycin (8639) and 25 ml of cycloheximide (8724). Mix gently and dispense into sterile Petri dishes or sterile bottles.


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 1031
Mono-Plates, 100x15 mm 10/pkg 1031M
Bottles, 2 oz 20/pkg 5042
     
Defibrinated Sheep Blood 100 ml 4147
     
Antibiotics    
Chloramphenicol, 10 ml 10/pkg 8629
Gentamycin, 10 ml 10/pkg 8639
Cycloheximide, 5 ml 10/pkg 8724

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