The first proctodeal (P1) segment in the hindgut of certain higher termites shows high alkalinity. We examined the bacterial diversity of the alkaline P1 gut segments of four species of higher termites by T-RFLP and phylogenetic analyses based on PCR-amplified 16S rRNA genes. The bacterial community of the P1 segment was apparently different from that of the whole gut in each termite. Sequence analysis revealed that Firmicutes (Clostridia and Bacilli) were dominant in the P1 segments of all four termites; however, the phylogenetic compositions varied among the termites. Although some of the P1 segment-derived sequences were related to the sequences previously reported from the alkaline digestive tracts of other insects, most of them formed phylogenetic clusters unique to termites. Such "termite P1 clusters" were distantly related to known bacterial species as well as to sequences reported from alkaline environments in nature. We successfully obtained enrichment cultures of Clostridia- and Bacilli-related bacteria, including putative novel species under anaerobic alkaline conditions from the termite guts. Our results suggest that the alkaline gut region of termites harbors unique bacterial lineages and are expected to be a rich reservoir of novel alkaliphiles yet to be cultivated.

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