Isolation and characterization of methylotrophic methanogens from anoxic marine sediments in Skan Bay, Alaska:: description of Methanococcoides alaskense sp nov., and emended description of Methanosarcina baltica

Three novel strains of methylotrophic methanogens were isolated from Skan Bay, Alaska, by using anaerobic cultivation techniques. The water was 65 m deep at the sampling site. Strains AK-4 (=OCM 774), AK-5(T) (=OCM 775(T)=DSM 17273(T)) and AK-9 (=OCM 793) were isolated from the sulfate-reducing zone of the sediments. Each of the strains was a non-motile coccus and occurred singly. Cells grew with trimethylamine as a catabolic substrate and strain AK-4 could also catabolize methanol. Yeast extract and trypticase peptones were not required for growth, but their addition to the culture medium slightly stimulated growth. Each of the strains grew at temperatures of 5-28 degrees C; they were slight halophiles and grew fastest in the neutral pH range. Analysis of the 16S rRNA gene sequences indicated that strain AK-4 was most closely related to Methanosarcina baltica. DNA-DNA hybridization studies showed 88% relatedness, suggesting that strain AK-4 represents a novel strain within this species. Strains AK-5(T) and AK-9 had identical 16S rRNA gene sequences that were most closely related to the sequence of Methanococcoides burtonii (99(.)8% sequence similarity). DNA-DNA hybridization studies showed that strains AK-5T and AK-9 are members of the same species (88% relatedness value), but strain AK-5T had a DNA-DNA relatedness value of only 55% to Methanococcoides burtonii. This indicates that strains AK-5(T) and AK-9 should be considered as members of a novel species in the genus Methanococcoides. We propose the name Methanococcoides alaskense sp. nov., with strain AK-5(T) (=OCM 775(T)=DSM 17273(T)) as the type strain.