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Two different isolation methods, the dilution colony-counting method (colony-isolation) and enrichment culture, were used to isolate sulfate-reducing bacteria (SRBs) from estuarine sediment in Japan. Lactate was used as an electron donor for colony-isolation, and lactate or propionate was used for enrichment culture. All isolates were classified into six different phylogenetic groups according to the 16S rRNA gene-based analysis. The closest relatives of the colony-isolates (12 strains) were species in the genera of Desulfobacterium, Desulfofrigus, Desulfovibrio and Desulfomicrobium. The closest known relative of the lactate-enrichment isolates was Desulfovibrio acrylicus and that of the propionate-enrichment isolates was Desulfobulbus mediterraneus. All isolates were incompletely-oxidizing SRBs. Overall patterns of utilization of electron donors and acceptors, as well as fermentative substrates, differed depending on the affiliation of the strain. Furthermore, even if several strains used the same substrate, the growth rates were often significantly different depending on the strain. It was strongly suggested that various species of SRBs could coexist in the sediment by competing for common substrates as well as taking priority in favorable or specific substrates for each species and the community of SRBs should be able to oxidize almost all major intermediates of anaerobic decomposition of organic matter such as lower fatty acids, alcohols and H2 as well as amino acids. Thus, it was indicated by the phylogenetic and physiological analyses of the isolates that the SRB community composed of diverse lineages of bacteria living in anoxic estuarine sediment should be able to play an extensive role in the carbon cycle as well as the sulfur cycle of the earth.

Diversity of substrate utilization and growth characteristics of sulfate-reducing bacteria isolated from estuarine sediment in Japan