Clostridium estertheticum subsp. laramiense (Kalchayanand et al.) Spring et al.
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2. Under anaerobic conditions, withdraw 0.5 ml of #1017 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 with 0.5 ml each from the suspension. Streak two blood plates to check for purity.
4. Incubate tubes under an anaerobic atmosphere at 10°C for 10-14 days. Incubate one agar plate anaerobically, and one aerobically, for aerobic contamination check.
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 connected 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.
After 10-14 days, growth is evidenced by turbidity and gas formation in the broth. No growth should occur on the agar plate.
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Validation list no. 46. Int. J. Syst. Bacteriol. 43: 624-625, 1993.
Truper HG, De Clari L. Taxonomic note: Necessary correction of specific epithets formed as substantives (nouns) "in apposition". Int. J. Syst. Bacteriol. 47: 908-909, 1997.
Kalchayanand N, et al. Characteristics of psychrotrophic Clostridium laramie causing spoilage of vacuum-packaged refrigerated fresh and roasted beef. J. Food Prot. 56: 13-17, 1993.
Spring S, et al. Characterization of novel psychrophilic clostridia from an Antarctic microbial mat: description of Clostridium frigoris sp. nov., Clostridium lacusfryxellense sp. nov., Clostridium bowmanii sp. nov. and Clostridium psychrophilum sp. nov. and reclassification of Clostridium laramiense as Clostridium estertheticum subsp. laramiense subsp. nov.. Int. J. Syst. Evol. Microbiol. 53: 1019-1029, 2003. PubMed: 12892121