Brain Heart Infusion Agar with Horse Blood and Taurocholate (BHI-HT Agar) is an enriched, non-selective medium used for the isolation and cultivation of a wide variety of bacteria, including yeasts and molds. The basic nutritive properties are brain heart infusion from solids as well as meat peptones, with the addition of yeast extract. Horse blood is added to stimulate growth. Sodium Taurocholate is added to help in the germination of Clostridium difficile spores. Clostridium difficile is not inhibited on the BHI-HT medium, and, when growing on this medium, will exhibit a characteristic colonial morphology, fluorescence, and smell. 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 w/ Horse Blood & Taurocholate
                            (BHI-HT) AS-6463

Item #	Description	Size	Price
AS-6463	BHI-HT mono plate	4 plates	10.55

 Formula

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

Yeast Extract, 5.0 g

Agar, 15.0 g

Distilled Water, 1000.0 ml

Horse Blood, 70.0 mL

Sodium Taurocholate, 1.0 g

  

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

Final weight 16.0 g +/- 1.6.

 

Precautions

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.

 

Procedure

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-HT 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 35-37^oC 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.

 

Limitations

BHI-HT 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	ATCC #	Results	Time (Hours)	Special Reaction
Bacteroides fragilis	25285	Growth	24
Enterococcus faecalis	29212	Growth	24
Clostridium sporogenes	3584	Growth	24
Clostridium beijerinckii	8260	Growth	24
Proteus mirabilis	12453	Growth	24
Clostridium perfringens	13124	Growth	24
Clostridium innocuum	14501	Growth	24
Clostridium sordellii	9714	Growth	24
Clostridium difficile	9689	Growth	24
Clostridium difficile	700057	Growth	24

 

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.

 

Organism	Expected Growth
B. fragilis	24 hrs
C. perfringens	24 hrs
C. difficile	24 hrs
S. aureus	24 hrs
E. coli	24 hrs

 

Physical Appearance:  BHI-HT agar should appear opaque burgundy red in color.

References

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.        Dowell, V. R., Jr., G. L. Lombard, F. S. Thompson and A. Y. Armfield.  1977.  Media for the Isolation, Characterization, and Identification of Obligately Anaerobic Bacteria.  USDHEW, CDC, Atlanta, GA 30333.

 

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

 

5.        Jousimies-Somer, H. R., P. Summanen, D. M. Citron, E. J. Baron, H. M. Wexler and S. M. Finegold.  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.

 

7.        George, W. L., V.L. Sutter, D. Citron, S. Finegold.  1979.  Selective and Differential Medium for Isolation of Clostridium difficile.  Journal of Clinical Microbiology.  9:214-219.

 

8.        Wilson KH, Kennedy MJ, Fekety FR:  Use of Sodium Taurocholate to Enhance Spore Recovery on a Medium Selective for Clostridium difficile.  Journal of Clinical Microbiology 1982, 15(3): 443-446.

 

9.        Marler LM, Siders JA, Wolters LC, Pettigrew Y, Skitt BL, Allen SD:  Comparison of Five Cultural Procedures for Isolation of Clostridium difficile from Stools.  Journal of Clinical Microbiology 1992, 30(2): 514-516.

 

10.     Edelstein, Martha. “Isolation and Identification of Clostridium difficile; Tissue Culture and Cytotoxicity Assay.”  Clostridium difficile: Its Role in Intestinal Disease.  Eds. Rolfe RD, Finegold SM.  San Diego:  Academic Press Inc, 1988.  287-307.