Bifidobacterium Selective Agar (BIFIDO) is a selective medium for the isolation and enumeration of Bifidobacterium species. BIFIDO contains Reinforced Clostridial Agar as the basal medium and Polymixin, Kanamycin, and Nalidixic acid as selective agents. The differential compounds of iodoacetate and 2, 3, 5 – triphenyltetrazolium chloride are also added. This medium is prepared, dispensed and packaged under oxygen-free conditions to prevent the formation of oxidized products prior to use.

               Anaerobe Systems PRAS Bifidobacterium
                  Selective Agar (BIFIDO) AS-6423

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Formula

Reinforced Clostridial Medium, 38.0 g

Agar, 15.0 g

Nalidixic Acid, 0.02 g

Polymyxin B Sulfate, 0.0085 g

Kanamycin Sulfate, 0.05 g

Iodoacetic Acid, 0.025 g

2,3,5-triphenyltetrazolium Chloride, 0.025 g

Distilled Water, 1000.0 ml

 

Final pH 7.1 +/- 0.2 at 25 degrees C.

Final weight 20.0 g +/- 2.0 for AS-6823.

Final weight 16.0 g +/- 1.6 for AS-6423.                                     

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:  BIFIDO 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 24-48 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

BIFIDO will not provide complete information for identification of bacterial isolates.  Additional test procedures and media are required for complete identification.  It is recommended that a non-selective medium such as Anaerobic Brucella Blood Agar also be inoculated from clinical specimens to assure growth of all species present.  Consult reference materials for additional information.

Quality Control

This medium should support good growth of some of the Bifidobacterium spp. found in clinical infections.

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

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.

Physical Appearance:  BIFIDO should appear opaque to translucent yellow/pink 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.     Engelkirk, P. G., Duben-Engelkirk, J., Dowell, V. R. 1992. Principles of Practices of Clinical Anaerobic Bacteriology. Star Publishing Co., Belmont, CA 94002. 

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. Wasdsworth – KTL Anaerobic Bacteriology Manual. Star Publishing Co., Belmont, CA 94002. 

5.     Munoa, F. J. and Pares, P. 1988. Selective Medium for Isolation and Enumeration of Bifidobacterium spp. Applied and Environmental Microbiology 54: 1715-1718. 

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.