Prevotella heparinolytica (Okuda et al.) Shah and Collins

1. Open vial according to enclosed instructions.
2. Under anaerobic conditions, withdraw 0.5 mL of Anaerobe Systems PRAS Chopped Meat medium from a single test tube (5 to 6 mL) and rehydrate the vial contents.
3. Aseptically transfer this aliquot back into the broth tube.  A slant and a pre-reduced blood plate may also be inoculated with 0.1 mL each of the cell suspension.  An aerobic blood plate may also be streaked to check for purity.
4. Incubate tubes and plate under anaerobic conditions at 37°C.  Incubate blood plate aerobically at 37°C.
5. Within 24 to 48 hours, growth should be evident by turbidity in the broth.  On agar surfaces, colonies are entire, glistening, circular, smooth, small, and raised. No growth should occur on the blood agar plate incubated aerobically.

ANAEROBIC CONDITIONS:   

Anaerobic conditions for transfer may be obtained by either of the following:

• Use of an anaerobic gas chamber, or                                         
• Placement of test tubes under a gassing cannula system hooked to anaerobic gas.   

Anaerobic conditions for incubation may be obtained by any of the following:

• Loose screw caps on test tubes in anaerobic chamber,                       
• Loose screw caps on test tubes in an activated anaerobic gas pack jar, or   
• Use of sterile butyl rubber stoppers on test tubes so that an anaerobic gas headspace is retained.   

Always use freshly prepared pre-reduced media or pre-reduced media that has been previously prepared but stored under anaerobic conditions. Resazurin in the media is a color indicator for anaerobic conditions. Observance of pink color in medium before use or during incubation shows anaerobic conditions have not been met and oxidation has occurred. Medium should be discarded.

Additional information on this culture is available on the ATCC web site at www.atcc.org.

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