Clostridium halophilum Fendrich 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 test tube (5 to 6 ml) and rehydrate the entire 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. 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.
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Fendrich C, et al. Clostridium halophilum new speices and Clostridium litorale new speices, an obligate halophilic and a marine species degrading betaine in the Stickland reaction. Arch. Microbiol. 154: 127-132, 1990.
Deed RW, et al. Validation list no. 39. Int. J. Syst. Bacteriol. 41: 580-581, 1991. PubMed: 1742202