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Brain Heart Infusion Agar (BHI Agar)

Brain Heart Infusion Agar (BHI Agar) is an enriched non-selective medium for the isolation and cultivation of most anaerobic bacteria and other fastidious microorganisms.  The basic nutritive properties are brain heart infusion from solids as well as meat peptones,with the addition of yeast extract.  This medium is supplemented with hemin and vitamin K1 as growth factors for most anaerobic bacteria.  This medium is prepared, dispensed, stored and packaged under oxygen-free conditions to prevent the formation of oxidized products prior to use.

                       Anaerobe Systems PRAS Brain
               Heart Infusion Agar (BHI) AS-6426

Brain Heart Infusion Agar Products

 Item #    





 BHI mono plate

 4 plates




Brain Heart Infusion, 37.0 g

Yeast Extract, 5.0 g

Hemin (0.1% Sol’n), 5.0 ml

Vitamin K1 (1% Sol’n), 0.05 ml

Agar, 15.0 g

Distilled Water, 1000.0 ml

L-Cysteine, 0.5 ml


Final pH 7.2 +/- 0.2 at 25 degrees C.

Final weight 16.0 g +/- 1.6.


For IN VITRO DIAGNOSTIC USE only.  Approved biohazard precautions and aseptic techniques should be observed when using this product.  This product is for use only by properly-trained and qualified personnel.  Sterilize all biohazard waste prior to disposal.

Storage and Shelf Life

Storage:  Upon receipt, store at room temperature (13°C-27°C) in original container until use.  Avoid overheating or freezing.  Do not use medium if there are signs of deterioration (shrinking, cracking or discoloration due to oxidation of media) or contamination. The expiration date applies to the product in its original packaging and stored as directed.  Do not use product past the expiration date shown on the container.

Shelf Life:  90 days from date of manufacture.


Specimen Collection:  Specimens for anaerobic culture should be protected from air (oxygen) during collection, transport and processing.  Consult appropriate references for detailed instructions concerning collection and transport of anaerobes. 

Methods for Use:  BHI AGAR should be inoculated directly with clinical material or a broth that has been previously inoculated from clinical material.  Inoculated plates should be streaked to obtain isolated colonies, immediately placed in an anaerobic atmosphere and incubated at 35-37oC for 18 – 48 hours.  Additional periods of incubation may be necessary to recover some anaerobes.  Detailed instructions for processing anaerobic cultures can be found in the appropriate references.

Materials Required But Not Provided

Standard microbiological supplies and equipment such as loops, saline blanks, slides, staining supplies, microscope, incinerator / autoclave, incubators, anaerobic chamber, other culture media and serological and biochemical reagents.


BHI AGAR will not provide complete information for identification of bacterial isolates.  Additional test procedures and media are required for complete identification.  Consult reference materials for additional information.

Quality Control

This medium should support good growth of many fastidious and non-fastidious anaerobes isolated from clinical specimens.

The following organisms are routinely used for quality control testing at Anaerobe Systems.

Organism Tested



Time (Hours)

Bacteroides fragilis



24 h

Prevotella melaninogenica



24-48 h

Fusobacterium mortiferum



24-48 h

Fusobacterium necrophorum



24-48 h

Clostridium perfringens



24 h

Clostridium sporogenes



24 h

Peptostreptococcus anaerobius



24 h

Proteus mirabilis



24 h

Propionibacterium acnes     or  Clostridium difficile





24 – 48 h

24 h

Staphylococcus aureus     or  Enterococcus faecalis





24 h

24 h

Escherichia coli



24 h


User Quality Control: The final determination to the extent and quantity of user laboratory quality control must be determined by the end user. 

If sterility testing is to be performed on this product, the appropriate percentage of the original shipment amount should be incubated anaerobically and aerobically for 48 – 96 hours.

If the nutritive/inhibitory capacity of this medium is to be tested for performance, it is recommended that the following ATCC organisms be evaluated for growth/inhibition.



Expected Growth

B. fragilis

24 hrs

C. perfringens

24 hrs

P. anaerobius

24 hrs

S. aureus

24 hrs

E. coli

24 hrs


Physical Appearance: BHI AGAR should appear opaque to translucent and yellow in color.



1.     Dowell, V. R., Jr. and T. M. Hawkins. 1974. Laboratory Methods in Anaerobic Bacteriology. CDC         Laboratory Manual. USDHEW C. D. C. Atlanta, GA 30333. 

2.     Dowell , V. R. Jr., and G. L. Lombard. 1977. Presumptive Identification of Anaerobic Non-sporeforming Gram-negative Bacilli. USDHEW, CDC. Atlanta, GA 30333. 

3.     Holdeman, L. V., F. P. Cato, and W. E. C. Moore. 1977. Anaerobe Laboratory Manual. Virginia Polytechnic Institute and State University. Blacksburg, VA 24061 

4.     Engelkirk, P. G., Duben-Engelkirk, J. and Dowell, V. R. 1992. Principles and Practices of Clinical Anaerobic Bacteriology. Star Publishing Co., Belmont, CA 94002. 

5.     Jousimies-Somer, H. R., Summanen, P., Citron, D. M., Baron, E. J., Wexler, H. M. and Finegold, S. M. 2002. Wadsworth – KTL Anaerobic Bacteriology Manual. Star Publishing Co., Belmont, CA 94002. 

6.     NCCLS. Quality Control for Commercially Prepared Microbiological Culture Media; Approved Standard- Third Edition. (2004). NCCLS document M22-A3. NCCLS, 940 West Valley Road, Suite 1400, Wayne, PA 19087-1898.