Thermoplasma acidophilum Darland et al.
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2. Add 1.5 ml of #569 broth to the vial. Make serial dilutions from this suspension by passing 0.5 ml into a tube containing 4.5 ml of the same medium. Repeat this procedure for a total of 7 to 8 passages.
3. Incubate the tubes at 55-60°C. Thermoplasma acidophilum requires oxygen, so slant the rack of tubes to provide greater surface area.
4. Growth appears in the first few tubes in 3 to 5 days. Subsequent transfers grow more rapidly, and should be passaged every 2 to 3 days. The inoculum size is 10 to 20%. Once stationary phase is reached, there is a drastic loss in viability. Glassware should be free of any trace of detergent or soap. Cells do not tolerate refrigeration.
5. For long term storage, freeze in liquid nitrogen. It is recommended that actively growing cells are harvested by centrifugation at 9000 rpm for 30 minutes, the supernatant removed, and the pellet resuspended in a smaller amount of fresh growth medium with an equal volume of 20% glycerol (10% F.C.) added. Small amounts of this suspension are aliquoted into small cyrovials and immediately placed in liquid nitrogen storage.
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Darland G, et al. A thermophilic, acidophilic mycoplasma isolated from a coal refuse pile. Science 170: 1416-1418, 1970. PubMed: 5481857
Skerman VB, et al. Approved lists of bacterial names. Int J Syst Bacteriol 30: 225-420, 1980.
Yasuda M, et al. Morphological variation of new Thermoplasma acidophilum isolates from Japanese hot springs. Appl. Environ. Microbiol. 61: 3482-3485, 1995. PubMed: 7574659