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BAM Media M24: Brain Heart Infusion (BHI) Broth and Agar

January 2001

Bacteriological Analytical Manual

Brain Heart Infusion (BHI) Broth and Agar (Equivalent media are available from Difco and BBL)


Medium 1

Calf brain, infusion from200 g
Beef heart, infusion from250 g
Proteose peptone (Difco) or
polypeptone (Bioquest)
10 g
NaCl5 g
Na2HPO4*2.5 g
Dextrose2.0 g
Distilled water1 liter


Dissolve ingredients of Medium 1 in distilled water with gentle heat.


*Difco does not specify waters of hydration.


Medium 2

Brain heart-infusion6.0 g
Peptic digest of animal tissue6.0 g
NaCl5.0 g
Dextrose3.0 g
Pancreatic digest of gelatin14.5 g
Na2HPO4**2.5 g
Distilled water1 liter


Suspend ingredients of Medium 2 in distilled water and boil for 1 min to completely dissolve.


**BBL does not specify waters of hydration.


For both Medium 1 and Medium 2, dispense broth into bottles or tubes for storage. Autoclave 15 min at 121°C. Final pH, 7.4 ± 0.2. Commercially available BHI is acceptable.

To prepare brain heart infusion agar, add 15 g agar to 1 liter BHI broth. Heat to dissolve agar before dispensing into bottles or flasks. Autoclave 15 min at 121°C.

For use with halophilic Vibrio spp., add NaCl to a final concentration of 2-3%.

Original Source: Bacteriological Analytical Manual, 8th Edition, Revision A, 1998.