Clostridium novyi (Migula) Bergey et al.
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ATCC highly recommends that appropriate personal protective equipment is always used when handling vials. For cultures that require storage in liquid nitrogen, it is important to note that some vials may leak when submersed in liquid nitrogen and will slowly fill with liquid nitrogen. Upon thawing, the conversion of the liquid nitrogen back to its gas phase may result in the vial exploding or blowing off its cap with dangerous force creating flying debris. Unless necessary, ATCC recommends that these cultures be stored in the vapor phase of liquid nitrogen rather than submersed in liquid nitrogen.
2. Under anaerobic conditions, withdraw 0.5 ml of recommended broth from a single tube (5 to 6 ml) and rehydrate the vial contents.
3. Aseptically transfer this aliquot back into the broth. Additional tubes may be inoculated with 0.5 ml each from the suspension. A slant can be inoculated with 0.3 ml. Inoculate several blood agar plate to check for colonial morphology and purity.
4. Incubate tubes and one blood plate under anaerobic conditions at 37oC. Incubate the second plate aerobically as a contamination check.
5. Within 24 to 48 hours, growth is evident by good turbidity and gas in the broth, and gas production is evident on the agar slant. No growth should occur on the blood agar plate incubated aerobically.
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.
Additional information on this culture is available on the ATCC web site at www.atcc.org.
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Skerman VB, et al. Approved lists of bacterial names. Int J Syst Bacteriol 30: 225-420, 1980.