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Brain Heart Infusion Broth (BHI)

Brain Heart Infusion Broth (BHI) is an enriched non-selective medium intended for the cultivation of most anaerobic bacteria and other fastidious microorganisms.  This medium is used in the inoculum preparation for antimicrobial susceptibility testing, is especially useful as a base for blood cultures and is also used in the broth-disc antimicrobial test procedure as described by Wilkins and Theil. BHI is an enriched non-selective broth medium that is useful in the cultivating of fastidious and non-fastidious microorganisms.  This medium will also support the growth of aerobic microorganisms from a variety of clinical and non-clinical specimens.  The basal medium is infusion from brains and beef heart and supplemented with vitamin K1 and hemin as growth factors for most anaerobes.  This medium is prepared, dispensed, stored and packaged under oxygen-free conditions to prevent the formation of oxidized products prior to use.  This medium contains resazurin as a redox indicator which turns pink upon significant oxygen exposure.

     Anaerobe Systems PRAS Brain Heart
             Infusion Broth (BHI) AS-872

Brain Heart Infusion Broth Products

Item #


BHI, 5mL

10 Tubes/pack



Brain Heart Infusion, 37.0 g

                Calf Brain Infusion, 7.7 g

                Beef Heart Infusion, 9.8 g

                Proteose Peptone, 10.0 g

                Dextrose, 2.0 g

                Sodium Chloride, 5.0 g

                Disodium Phosphate, 2.5 g

Hemin, .005 g

Vitamin K1, .002 g

Resazurin, .001 g

Yeast Extract, 5.0 g

L-Cysteine, 0.5 g

Distilled Water, 1000.0 ml


Final pH 7.3 +/- 0.3 at 25 degrees C.

Final volume 5.0 ml +/- 0.5.


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 packaging until use.  Avoid overheating or freezing.  Do not use medium if there are signs of deterioration (discoloration or evaporation) 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:  1 year from date of manufacture.


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

Methods for Use:  BHI should be inoculated directly with clinical material.  Inoculated tubes should be 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, pipets, saline blanks, slides, staining supplies, microscope, incinerator/autoclave, incubators, anaerobic chamber, other culture media and serological and biochemical reagents.


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

Quality Control

If used properly, 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 assurance performance testing at Anaerobe Systems. 

Organism Tested




Time (Hours)

Bacteroides fragilis



24 h

Prevotella melaninogenica



24 – 48 h

Bacteroides vulgatus



24 h

Fusobacterium nucleatum



24 – 48 h

Fusobacterium necrophorum



24 – 48 h

Clostridium perfringens



24 h

Clostridium novyi



24 – 48 h

Peptostreptococcus anaerobius



24 h

Clostridium difficile



24 h

Staphylococcus aureus



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 capacity of this media is to be tested for performance, it is recommended that the following ATCC organisms be evaluated for growth.






B. fragilis

24 hrs

P. melaninogenica

48 hrs

C. perfringens

24 hrs

F. necrophorum

48 hrs

S. aureus

24 hrs

 Physical Appearance:  BHI should appear as a clear, yellowish liquid within the 16mm x 100mm glass tube.


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

 5.        Wilkins, T. D. and T. Theil.  1973.  Modified Broth-Disc Method for Testing the Antibiotic Susceptibility of Anaerobic Bacteria.  Antimicrob. Agents Chemother. 3: 350 – 356.

 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.

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