Clostridium frigoris Spring et al.
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2. Open the frozen vial and immediately place the vial under a stream of sterile gas, to maintain anaerobicity.
3. Under anaerobic conditions, withdraw the cell suspension from the vial and transfer it to a single test tube of ATCC #1490 broth.
4. A second 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 4°C. Incubate aerobic plate(s) and broth tube(s) at 30oC.
6. Within 9 to 12 days, growth is evident by turbidity in broth and in the inoculum at the base of the slant. No growth should occur on blood agar plates 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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Brambilla E, et al. 16S rDNA diversity of cultured and uncultured prokaryotes of a mat sample from Lake Fryxell, McMurdo Dry Valleys, Antarctica. Extremophiles 5: 23-33, 2001. PubMed: 11302499
microbial mat sample from shallow, moated area, around Lake Fryxell, an antarctic freshwater lake, McMurdo Dry Valleys, Antarctica
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