Clostridium formicaceticum Andreesen et al.

1. This culture must be propagated under strictly anaerobic, conditions (see below).
2. Open the vial according to enclosed instructions.
3. Under anaerobic conditions, withdraw 0.5 mL of #591 broth from a single test tube and rehydrate the entire vial contents.
4. Aseptically transfer this aliquot back into the broth tube.  A slant and a pre-reduced blood plate may be inoculated with 0.1 mL each of the cell suspension.  Also inoculate a non-selective plate and tube of broth to test for the presence of aerobic contaminates.
5. Incubate tubes and plate under anaerobic conditions at 37°C.  Incubate aerobic plate(s) and broth tube(s) at 37°C.
6. In 1 to 3 days, growth should be evident by light turbidity in the broth and by turbidity in the liquid at the base of the slant culture.  On #591 plates, colonies are entire, glistening, circular, smooth, and opaque.  Cells are long, regular rods.  Many cells exhibit motility.  No growth should occur on the aerobic plate.

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.

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