Deltaproteobacteria sp.

1. Open vial anaerobically according to enclosed instructions.
2. Under anaerobic conditions, withdraw 0.5 mL of recommended broth from a single test tube (5 to 6 mL) and rehydrate the entire vial contents.
3. Aseptically transfer this aliquot back into the broth tube.  Additional tubes may be inoculated from this original tube.  Streak two blood plates to check for purity;no growth should occur anaerobically or aerobically.
4. Incubate tubes under an anaerobic atmosphere at 28°C.  Incubate one agar plate anaerobically and one aerobically at 28°C.
5. This organism will take several weeks to achieve good growth. Under optimal conditions the generation time is 32 hours. Growth is detected either by microscopic inspection or by measuring the reduction of arsenate. Cells are Gram-negative motile rods.

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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