Bacteroides Vulgatus Selective Agar (BVSA) is an enriched selective and differential medium for the isolation and presumptive identification of obligately anaerobic gram-negative bacilli of the Bacteroides fragilis group. BVSA contains kanamycin, vancomycin and colistin at concentrations that inhibit most facultative anaerobes. BVSA also contains bile, which inhibits anaerobic gram-negative rods except the B. fragilis group and other bile resistant Bacteroides and Fusobacteria.  Esculin in the medium permits recognition of esculin hydrolysis by organisms which produce a brown to black coloration in the media around the colonies.  Hemin is added as a growth factor.  This medium is prepared, dispensed and packaged under oxygen-free conditions to prevent the formation of oxidized products prior to use.

   Anaerobe Systems PRAS Bacteroides Vulgatus
             Selective Agar (BVSA) AS-6425

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Formula

Pancreatic Digest of Casein, 10.0 g

Soy Peptone, 3.0 g

Sodium Chloride, 5.0 g

Agar, 15.0 g

Yeast Extract, 2.0 g

Oxgall, 20.0 g

Esculin, 1.0 g

Ferric Ammonium Citrate, 0.5 g

Hemin (0.1% Soln), 10.0 ml

Vitamin K₁ (1% Soln), 1.0 ml

Kanamycin, 200.0 mg

Vancomycin, 7.5 mg

Colistin, 0.015 mg

Distilled Water, 1000.0 ml

 

Final pH 7.0 +/- 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:  BVSA 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; however, extended incubation may result in the loss of selectivity of the medium resulting in overgrowth of organisms, which should be inhibited.  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 or autoclave, incubators, anaerobic chamber or anaerobic jars, disinfectant, other culture media and serological and biochemical reagents.

Limitations

BVSA 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.  Some strains of facultative organisms, which might be inhibited, may grow on BVSA.  A test for aerotolerance should be used to confirm that each colony type is an obligate anaerobe.  Consult reference materials for additional information.

Quality Control

This medium should inhibit the growth of most facultative anaerobes and most obligately anaerobic bacteria other than Bacteroides.  Members of the B. fragilis group should grow as brown to black colonies surrounded by a brown zone in the medium, with the exception of B. vulgatus.

The following organisms are routinely used for quality assurance performance 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.