Methylosinus sporium (ex Romanovskaya et al.) Bowman et al.
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2. Using a single tube of broth, aseptically transfer 0.5 mL to the vial and rehydrate the entire pellet.
3. Aseptically transfer this aliquot back to the broth tube and mix well. Transfer 0.1 mL to slants. Plate the rehydrated culture (0.1 mL) onto non-selective medium (to test for purity). When using ATCC Medium #1306 use a gas mixture of 50% methane and 50% air. With ATCC Medium #784 no special gas is required.
4. Incubate the culture at 30°C for 48 to 72 hours. When using ATCC Medium #1306 the culture needs to be fed a mixture of 50% methane and 50% air every 24 hours for best results.
5. Growth should be detected within 3 to 5 days. No growth should be detected on the non-selective plates.
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Whittenbury R, et al. Enrichment, isolation and some properties of methane-utilizing bacteria. J Gen Microbiol 61: 205-218, 1970. PubMed: 5476891
Bowman JP, et al. Revised taxonomy of the methanotrophs: Description of Methylobacter gen. nov., emendation of Methylococcus, validation of Methylosinus and Methylocystis species, and aproposal that the family Methylococcaceae includes only the Group I methanotrophs. Int. J. Syst. Bacteriol. 43: 735-753, 1993.