BRAIN HEART INFUSION (BHI) AGAR
| Cat. no. L35 | BHI Agar, 20x125mm Tube, 10ml Slant | 20 or 100 tubes/box |
| Cat. no. W15 | BHI Agar, 15x100mm Plate, 26ml | 10 plates/bag |
| Cat. no. X10 | BHI Agar, 50ml HardyFlask™, 12ml Slant | 20 flasks/box |
| Cat. no. A20 | BHI Agar with Blood, 15x100mm Plate, 18ml | 10 plates/bag |
| Cat. no. W185 | BHI Agar with Blood, 25x100mm Plate, 37ml | 5 plates/bag |
| Cat. no. X13 | BHI Agar with Blood, 50ml HardyFlask™, 12ml | 20 flasks/box |
| Cat. no. L31 | BHI Agar with Blood, 20x125mm Tube, 10ml Slant | 20 or 100 tubes/box |
| Cat. no. X12 | BHI Agar with Blood and Gentamicin, 50ml HardyFlask™, 12ml |
20 flasks/box |
| Cat. no. W65 | BHI Agar with Blood, Chloramphenicol and Gentamicin, 15x100mm Plate, 26ml |
10 plates/bag |
| Cat. no. X14 | BHI Agar with Blood, Chloramphenicol and Gentamicin, 50ml HardyFlask™, 12ml |
20 flasks/box |
| Cat. no. L33 | BHI Agar with Chloramphenicol and Cycloheximide, 20x125mm Tube, 10ml |
20 or 100 tubes/box |
| Cat. no. X11 | BHI Agar with Chloramphenicol and Cycloheximide 50ml HardyFlask™, 12ml |
20 flasks/box |
| Cat. no. W186 | BHI Agar with Chloramphenicol and Gentamicin, 25x100mm Plate, 37ml |
5 plates/bag |
| Cat. no. K07 | BHI with Agar and Fildes, 16x100mm Tube, 6ml Deep | 20 or 100 tubes/box |
INTENDED USE
Hardy Diagnostics Brain Heart Infusion (BHI) Agar is a general purpose nutrient medium recommended for the cultivation and isolation of a variety of microorganisms, including bacteria, yeasts, and molds. Media enriched with Fildes is useful for cultivating capsular strains of Haemophilus influenzae. The addition of defibrinated sheep blood and antimicrobics produces a selective medium used for the isolation of pathogenic fungi from specimens heavily contaminated with bacteria and saprophytic fungi.(2)
SUMMARY
Rosenow, by adding brain tissue to dextrose broth, discovered a medium useful in the cultivation of streptococci.(8) Formula modifications were made by various researchers who found the medium effective in the recovery of dental pathogens.(4,5,8) The medium was further modified by the addition of agar and a variety of supplements and enrichments which further enhanced the recovery of microorganisms.
The brain heart infusion, peptone and dextrose components of the medium provide the nutrients to BHI Agar. Organic nitrogen, carbon, sulfur, vitamins and trace substances are provided by the peptones and infusion. Dextrose provides the carbohydrate source for fermentative microorganisms. Disodium phosphate is added to the medium in order to maintain an optimal pH. Additionally, a variety of supplements has been added to further enhance the recovery of fastidious microorganisms.
BHI with Agar and Fildes is a semi-solid medium 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)
BHI Agar with Blood is used for the recovery of fungal species.(13) Blood provides essential growth factors for the more fastidious fungal organisms.
BHI Agar with Blood, Gentamicin and Chloramphenicol is used for the selective isolation of fungi from specimens heavily contaminated with bacteria.(2) Gentamicin inhibits the growth of gram-negative and some gram-positive bacteria. A wide range of gram-positive and gram-negative bacteria are inhibited by chloramphenicol, a broad-spectrum antimicrobic.
BHI Agar with Chloramphenicol and Cycloheximide is used for the selective isolation of pathogenic fungi. Cycloheximide inhibits saprophytic fungi while Chloramphenicol acts as an antibacterial agent. The incorporation of both antimicrobics achieves more complete selectivity against growth of saprophytic fungi and bacteria.(2)
FORMULA
Ingredients per liter of deionized water:*
| BHI Agar: | |
| Pancreatic Digest of Casein | 16.0gm |
| Brain Heart Infusion from Solids | 10.0gm |
| Peptic Digest of Animal Tissue | 5.0gm |
| Sodium Chloride | 3.0gm |
| Disodium Phosphate | 2.5gm |
| Dextrose | 2.0gm |
| Agar | 13.5gm |
| BHI Agar with Blood: | |
| In addition to the above formulation, contains: | |
| Sheep Blood | 50.0ml |
| BHI Agar with Blood and Gentamicin: | |
| In addition to the above formulation, contains: | |
| Gentamicin | 5.0mg |
| Sheep Blood | 50.0ml |
| BHI Agar with Chloramphenicol and Gentamicin**: | |
| In addition to the above formulation, contains: | |
| Chloramphenicol | 50.0mg |
| Gentamicin | 5.0mg |
| BHI Agar with Blood, Chloramphenicol and Gentamicin**: | |
| In addition to the above formulation, contains: | |
| Chloramphenicol | 50.0mg |
| Gentamicin | 5.0mg |
| Sheep Blood | 50.0ml |
| BHI Agar with Chloramphenicol and Cycloheximide**: | |
| Pancreatic Digest of Casein | 16.0gm |
| Brain Heart Infusion | 8.0gm |
| Peptic Digest of Animal Tissue | 5.0gm |
| Sodium Chloride | 5.0gm |
| Disodium Phosphate | 2.5gm |
| Dextrose | 2.0gm |
| Cycloheximide | 0.5gm |
| Chloramphenicol | 0.05gm |
| Agar | 13.5gm |
| BHI with Agar and Fildes: | |
| Pancreatic Digest of Casein | 14.5gm |
| Brain Heart Infusion from Solids | 10.0gm |
| Peptic Digest of Animal Tissue | 5.0gm |
| Sodium Chloride | 3.0gm |
| Disodium Phosphate | 2.5gm |
| Dextrose | 2.0gm |
| Fildes Supplement | 20.0ml |
| Agar | 2.5gm |
Final pH 7.4 +/- 0.3 at 25 degrees C.
* Adjusted and/or supplemented as required to meet performance criteria.
** This media contains chloramphenicol and/or cycloheximide which are ingredients considered potentially hazardous.
Refer to the keyword "MSDS Introduction", in the Hardy Diagnostics software program HUGO™, for more information on handling potentially hazardous material.
STORAGE AND SHELF LIFE
Storage: Upon receipt store all BHI Agar products* at 2-8 degrees C. (*except Cat. no. L33, L35, X10 and X11). Products L33, L35, X10 and X11 may be stored at 2-30 degrees C. Products should not be used if there are any signs of contamination, deterioration (shrinking, cracking, or discoloration), hemolysis, contamination, or if the expiration date has passed. Product is light and temperature sensitive; protect from light, excessive heat and freezing.
The expiration date applies to the product in its intact packaging when stored as directed.
This product, depending on its formulation, has the following shelf life from the date of manufacture:
| 90 Days: | A20 | BHI Agar with Blood |
| W15 | BHI Agar | |
| W65 | BHI Agar with Blood, Chloramphenicol, and Gentamicin | |
| W185 | BHI Agar with Blood | |
| W186 | BHI Agar with Chloramphenicol, and Gentamicin | |
| 120 Days: | L31 | BHI Agar with Blood |
| X10 | BHI Agar | |
| X11 | BHI Agar with Chloramphenicol and Cycloheximide | |
| X12 | BHI Agar with Blood and Gentamicin | |
| X13 | BHI Agar with Blood | |
| X14 | BHI Agar with Blood, Chloramphenicol and Gentamicin | |
| 180 Days: | K07 | BHI with Agar and Fildes |
| 365 Days: | L33 | BHI Agar with Chloramphenicol and Cycloheximide |
| L35 | BHI Agar | |
Refer to the keyword "Storage", in the Hardy Diagnostics software program HUGO™, for more information on storing culture media.
PRECAUTIONS
This product is for in vitro diagnostic use only and is to be used only by adequately trained and qualified laboratory personnel. Observe approved biohazard precautions and aseptic techniques. All laboratory specimens should be considered infectious and handled according to "standard precautions". The "Guideline for Isolation Precautions" is available from the Centers of Disease Control and Prevention at www.cdc.gov/ncidod/dhqp/gl_isolation.html.
For additional information regarding specific precautions for the prevention of the transmission of all infectious agents from laboratory instruments and materials, and for recommendations for the management of exposure to infectious disease, refer to CLSI document M29.
Sterilize all biohazard waste before disposal.
Refer to the keyword "Precautions", in the Hardy Diagnostics software program HUGO™, for more information regarding general precautions when using culture media.
Refer to the keyword "MSDS", in the Hardy Diagnostics software program HUGO™, for more information on handling potentially hazardous material.
PROCEDURE
Specimen Collection: Consult listed references for information on specimen collection.(1-3,6) 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.
Method of Use for Cultivation and Isolation of Bacteria: Consult listed references for the appropriate cultivation techniques using this medium.(1-3,6) Medium should be brought to room temperature and agar surface should be dry prior to inoculation. If the specimen to be cultured is on a swab, roll the swab over a small area of the agar surface. Streak for isolation with a sterile loop. Incubate plates aerobically at 35 +/- 2.0 degrees C. for 24 to 48 hours. Protect from light. Examine plates for colonial morphology.
Method of Use for Cultivation and Isolation of Fungi: A selective and non-selective medium should be inoculated for isolation of fungi from potentially contaminated specimens. Media should be incubated at room temperature (15-30 degrees C.) with increased humidity. Two sets of media should be inoculated for isolation of fungi causing systemic mycoses; one set incubated at 15-30 degrees C. and one set at 35 +/- 2.0 degrees C. Cultures should be examined weekly for fungal growth. Media should be held for four to six weeks before being reported as negative.
BHI Agar slants (without supplements or antimicrobics) are used primarily for the cultivation and maintenance of pure cultures of microorganisms.
INTERPRETATION OF RESULTS
Media should be examined for typical bacterial and fungal colonial morphology. Consult listed references for the interpretation of growth and other identification tests to identify growth of organism(s) on this media.(1-3,6)
LIMITATIONS
It is recommended that biochemical and/or serological tests be performed on colonies from pure culture for complete identification.(1-3,6)
Refer to the keyword "Limitations", in the Hardy Diagnostics software program HUGO™, for more information regarding general limitations on culture media.
MATERIALS REQUIRED BUT NOT PROVIDED
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.
QUALITY CONTROL
The following organisms are routinely used for testing at Hardy Diagnostics:
| Test Organisms | Inoculation Method* | Incubation | Results | ||
| Time | Temperature | Atmosphere | |||
| BHI Agar: | |||||
| Escherichia coli ATCC® 25922 |
A | 24-48hr | 35°C | Aerobic | Growth |
| Staphylococcus aureus ATCC® 25923 |
A | 24-48hr | 35°C | Aerobic | Growth |
| Staphylococcus epidermidis ATCC® 12228 |
A | 24-48hr | 35°C | Aerobic | Growth |
| BHI Agar with Blood: | |||||
| Streptococcus pyogenes ATCC® 19615 |
A | 18-24hr | 35°C | Aerobic | Growth; beta-hemolysis |
| Streptococcus pneumoniae ATCC® 6305 |
A | 18-24hr | 35°C | Aerobic | Growth; alpha-hemolysis |
| Aspergillus brasiliensis formerly A. niger ATCC® 16404 |
G | 5-7 days | 30°C | Aerobic | Growth |
| Staphylococcus aureus ATCC® 25923 |
A | 18-24hr | 35°C | Aerobic | Growth |
| Escherichia coli ATCC® 25922 |
A | 18-24hr | 35°C | Aerobic | Growth |
| BHI Agar with Blood and Gentamicin / BHI Agar with Chloramphenicol, and Gentamicin / BHI Agar with Blood, Chloramphenicol, and Gentamicin: | |||||
| Candida albicans ATCC® 10231 |
A | 18-24hr | 35°C | Aerobic | Growth |
| Trichophyton mentagrophytes ATCC® 9533 |
G | 7 days | 15-30°C | Aerobic | Growth |
| Escherichia coli ATCC® 25922 |
B | 18-24hr | 35°C | Aerobic | Partial to complete inhibition |
| Pseudomonas aeruginosa ATCC® 27853 |
B | 24hr | 35°C | Aerobic | Partial to complete inhibition |
| BHI Agar with Chloramphenicol and Cycloheximide: | |||||
| Candida albicans ATCC® 10231 |
A | 18-24hr | 35°C | Aerobic | Growth |
| Trichophyton mentagrophytes ATCC® 9533 |
G | 7 days | 15-30°C | Aerobic | Growth |
| Escherichia coli ATCC® 25922 |
B | 18-24hr | 35°C | Aerobic | Partial to complete inhibition |
| Aspergillus brasiliensis formerly A. niger ATCC® 16404 |
G | 7 days | 15-30°C | Aerobic | Partial to complete inhibition |
| BHI with Agar and Fildes: | |||||
| Haemophilus influenzae ATCC® 10211 |
D | 24hr | 35°C | Aerobic | Growth |
User Quality Control
Check for signs of contamination and deterioration. Users of commercially prepared media may be required to perform quality control testing with at least one known organism to demonstrate growth or a positive reaction; and at least one organism to demonstrate inhibition or a negative reaction (where applicable). Refer to the following keywords, in the Hardy Diagnostics software program HUGO™, for more information on QC: "Introduction to QC", "QC of Finished Product", and "The CLSI (NCCLS) Standard and Recommendations for User QC of Media". Also see listed references for more information.(1-3,6,10)
* Refer to the keyword "Inoculation Procedures", in the Hardy Diagnostics software program HUGO™, for a description of inoculation procedures.
PHYSICAL APPEARANCE
- BHI Agar, BHI Agar with Chloramphenicol and Gentamicin, and BHI Agar with Chloramphenicol and Cycloheximide should appear slightly opalescent, and light amber in color.
- BHI Agar with Blood; BHI Agar with Blood and Gentamicin; and BHI Agar with Blood, Chloramphenicol, and Gentamicin should appear opaque, and cherry to brick-red in color.
- BHI with Agar and Fildes should appear as a semi-solid medium that should appear clear, and dark amber in color.
Sterile plate of BHI Agar (Cat. no. W15).
Escherichia coli (ATCC® 25922) colonies growing on BHI Agar (Cat. no. W15). Incubated aerobically for 24 hours at 35 deg. C.
Uninoculated plate of BHI Agar with Blood (Cat. no. A20).
Candida albicans (ATCC® 10231) colonies growing on BHI Agar with Blood, Chloramphenicol and Gentamicin (Cat. no. W65). Incubated aerobically for 24 hours at 35 deg. C.
Streptococcus pneumoniae (ATCC® 6305) colonies growing on BHI Agar with Blood (Cat. no. A20). Incubated aerobically for 24 hours at 35 deg. C.
Escherichia coli (ATCC® 25922) growth inhibited on BHI Agar with Blood, Chloramphenicol and Gentamicin (Cat. no. W65). Incubated aerobically for 24 hours at 35 deg. C.
Trichophyton mentagrophytes (ATCC® 9533) colonies growing on BHI Agar with Blood, Chloramphenicol and Gentamicin (Cat. no. W65). Incubated aerobically for 96 hours at 30 deg. C.
Candida albicans (ATCC® 10231) colonies growing on BHI Agar with Chloramphenicol and Cycloheximide (Cat. no. L33). Incubated aerobically for 24 hours at 35 deg. C.
Escherichia coli (ATCC® 25922) growth inhibited on BHI Agar with Chloramphenicol and Cycloheximide (Cat. no. L33). Incubated aerobically for 24 hours at 35 deg. C.
REFERENCES
1. August, M.J., et al. 1990. Cumitech 3A; Quality Control and Quality Assurance Practices in Clinical Microbiology, Coordinating ed., A.S. Weissfeld. American Society for Microbiology, Washington, D.C.
2. Murray, P.R., et al. 1995. Manual of Clinical Microbiology, 6th ed. American Society for Microbiology, Washington, D.C.
3. Forbes, B.A., et al. 1998. Bailey and Scott's Diagnostic Microbiology, 10th ed. 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. 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. 1996. M22-A2, Vol. 16, No. 16. Clinical Laboratory Standards Institute (CLSI - formerly NCCLS), Villanova, PA.
11. Fildes, P. 1920. Br. J. Exp. Pathol.; 1:129.
12. Fildes, P. 1921. Br. J. Exp. Pathol.; 2:16.
13. Creitz and Puckett. 1954. J. Clin. Pathol.; 24:1318.
14. J. Chron. Dis.; 5:545, 1957.
15. J. Lab and Clin. Med.; 55:116, 1960.
ATCC is a registered trademark of the American Type Culture Collection.
102208mg