The microbial diversity of two types of methanogenic granular sludge, mesophilic (35 °C) and thermophilic (55 °C), which had been treating sucrose/propionate/acetate-based artificial wastewater were compared. 16S rDNA clone libraries were constructed by PCR with a prokaryote-specific primer set, and partial sequencing of the clonal 16S rDNAs was conducted for phylogenetic analysis. Of 115 mesophilic granule and 110 thermophilic granule clones sequenced, 19 and 22%, respectively, were phylogenetically affiliated with the domain and the remainder in each case were assigned to the domain Within the domain the 16S rDNA clones in both libraries showed relatively close relationships with those of methanogens. Within the a major group represented in the mesophilic clone library was the delta subclass of the (27%), in which high degrees of relatedness were observed between the clonal 16S rDNA sequences and those of previously identified syntrophic bacteria and sulfate-reducing bacteria. In contrast, in the thermophilic clone library, the group (19%), the green non-sulfur bacteria (18%) and the low G+C subclass of the Gram-positive bacteria (18%) were predominant. A significant difference between the two libraries was that no clone affiliated with the was detected in the thermophilic clone library, whereas the was the most predominant group in the mesophilic clones. Thirty-six and 24 different sequences were found in the mesophilic and thermophilic clones, respectively, suggesting that the microbial diversity of the thermophilic granule was lower than that of the mesophilic granule.


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