bhi broth

Cat. no. R10 BHI Broth, 13x100mm Tube, 0.5ml 20 tubes/box
Cat. no. R15 BHI Broth, 13x100mm Tube, 2ml 20 tubes/box
Cat. no. R20 BHI Broth, 13x100mm Tube, 5ml 20 tubes/box
Cat. no. K110 BHI Broth, 15x103mm Tube, 1ml 20 tubes/box
Cat. no. K27 BHI Broth, 15x103mm Tube, 5ml 20 tubes/box
Cat. no. K25 BHI Broth, 16x125mm Tube, 10ml 20 tubes/box
Cat. no. K08 BHI Broth with Fildes, 16x125mm Tube, 10ml 20 tubes/box


Hardy Diagnostics Brain Heart Infusion (BHI) Broth is a general purpose nutrient medium recommended for the cultivation and isolation of a variety of fastidious and nonfastidious microorganisms, including bacteria, yeast, and mold. Brain Heart Infusion (BHI) Broth enriched with Fildes is useful for cultivating capsular strains of Haemophilusinfluenzae.


Rosenow discovered a medium useful in the cultivation of streptococci by adding brain tissue to Dextrose Broth. (8) Formula modifications were made by various researchers who found the medium effective in the recovery of dental pathogens.(4,5,8)

The formulation presently in use employs calf brain, which is equivalent in nutritional value to beef heart infusion but provides a more optically clear medium. Additionally, a variety of supplements have been added to further enhance the recovery of fastidious microorganisms.

BHI with Fildes is used for organisms requiring blood derivatives, especially Haemophilus spp. Fildes is a peptic digest of sheep blood which is rich in X- and V-factors.(11,12)

Hardy Diagnostics BHI Broth, 0.5ml (Cat. no. R10) is used to prepare suspensions for microdilution minimal inhibitory concentration (MIC) inocula.


Ingredients per liter of deionized water:*

Calf Brain-Beef Heart Infusion 17.5gm
Pancreatic Digestion of Gelatin 10.0gm
Sodium Chloride 5.0gm
Disodium Phosphate 2.5gm
Dextrose 2.0gm

In addition, BHI Broth with Fildes (Cat. no. K08) contains:

Fildes Enrichment 50.0ml

Final pH 7.4 +/- 0.3 at 25ºC.

* Adjusted and/or supplemented as required to meet performance criteria.


Storage: Upon receipt store BHI Broth at 2-30ºC. away from direct light. BHI Broth, 0.5ml (Cat. no. R10) and BHI Broth with Fildes (Cat. no. K08) should be stored at 2-8ºC. away from direct light. Media should not be used if there are any signs of deterioration (shrinking, cracking, or discoloration), contamination, or if the expiration date has passed. Product is light and temperature sensitive; protect from light, excessive heat and freezing.



Specimen Collection: Infectious material should be submitted directly to the laboratory without delay and protected from excessive heat and cold. If there is to be a delay in processing, the specimen should be inoculated onto an appropriate transport medium and refrigerated until inoculation. Consult listed references for information on specimen collection.(1-3,6)

Method of Use: It is recommended that liquid media for anaerobic incubation should be reduced prior to inoculation by placing tubes (with loosened caps) under anaerobic conditions for 18-24 hours. Alternatively, the media may be reduced by bringing the media up to 100ºC. in a boiling waterbath. Loosen screw caps slightly before heating, and tighten during cooling to room temperature. Consult listed references for the appropriate cultivation techniques using this medium.(1-3,6)

Inoculate broth using aseptic techniques. Swab specimens may be inserted into the medium after plated media has been inoculated. Medium should be incubated at 35-37ºC., checking daily, as needed. Growth or turbidity should be confirmed by Gram stain and subculture onto an appropriate solid medium.


Consult listed references for the interpretation of growth and other biochemical tests needed to identify organisms grown in this medium.(1-3,6)



Standard microbiological supplies and equipment such as loops, other culture media, swabs, applicator sticks, incinerators, and incubators, etc., as well as serological and biochemical reagents, are not provided.


Test Organisms Inoculation Method* Incubation Results
Time Temperature Atmosphere
BHI Broth**:
Escherichia coli
ATCC ® 25922
A 18-24hr 35°C Aerobic Growth; broth becomes turbid
Staphylococcus aureus
ATCC ® 25923
A 18-24hr 35°C Aerobic Growth; broth becomes turbid
BHI Broth with Fildes:
Haemophilus influenzae
ATCC ® 10211
A 24-48hr 35°C Aerobic Growth; broth becomes turbid
Staphylococcus epidermidis
ATCC ® 12228
A 24-48hr 35°C Aerobic Growth; broth becomes turbid

** Refer to guidelines for testing in the CLSI publication M22-A.(10)

User Quality Control


E. coli growing in BHI Broth

Escherichia coli (ATCC ® 25922) growing in BHI Broth (Cat. no. R20). Incubated aerobically for 24 hours at 35ºC.

BHI Broth

Uninoculated tube of BHI Broth (Cat. no. R20).


1. Anderson, N.L., et al. Cumitech 3B; Quality Systems in the Clinical Microbiology Laboratory, Coordinating ed., A.S. Weissfeld. American Society for Microbiology, Washington, D.C.

2. Versalovic, J., et al. Manual of Clinical Microbiology. American Society for Microbiology, Washington, D.C.

3. Tille, P.M., et al. Bailey and Scott's Diagnostic Microbiology, C.V. Mosby Company, St. Louis, MO.

4. Falk, C.R., et al. 1939. J. Bacteriol.; 37:121.

5. Haden, R.L. 1932. Arch. Internal Med.; 32:828.

6. Isenberg, H.D. Clinical Microbiology Procedures Handbook, Vol. I, II & III. American Society for Microbiology, Washington, D.C.

7. MacFaddin, J.F. 1985. Media for Isolation, Cultivation, Identification, Maintenance of Bacteria, Vol. I. Williams & Wilkins, Baltimore, MD.

8. Hitchens, A.P. 1921. J. Infect. Disease; 29:390.

9. Rosenow, E.C. 1919. J. Dental Research; 1:205.

10. Quality Assurance for Commercially Prepared Microbiological Culture Media, M22. Clinical and Laboratory Standards Institute (CLSI - formerly NCCLS), Wayne, PA.

11. Fildes, P. 1920. Br. J. Exp. Pathol.; 1:129.

12. Fildes, P. 1921. Br. J. Exp. Pathol.; 2:16.

ATCC is a registered trademark of the American Type Culture Collection.