Samples of subgingival dental tissues were examined for the presence of methanogenic activities. Using enrichment cultures, methanogenic activities were detected in 9 of 17 individuals. A mesophilic, Gram-positive, irregular coccoid methanogen, which showed close resemblance to a Methanosarcina sp., was isolated from one sample collected from a patient with type IV periodontal pocket (the periodontal pocket is a space bounded by the tooth on one side and by ulcerated epithelium lining the soft tissue wall on the other). The isolate used methanol, methylamine, acetate, and H-2-CO2 as the sole source of carbon. However, the isolate was unable to use formate and trimethylamine as growth substrates. The organism had an optimum pH of 6.5 and an optimum temperature of 37degreesC. The isolate not only used ammonia, but also used nitrate as a nitrogen source. The niche of this methanogen in periodontal pockets may be to carry out terminal oxidation of simple organic compounds such as methanol and acetate produced by other obligate anaerobes present in periodontal pockets. This methanogen may also play a vital role in interspecies hydrogen transfer, as demonstrated by its use of H-2-CO2 as a substrate. The isolate produced significant amount of methane in vitro.