Abstract
BackgroundAlthough mice have long served as an animal model for periodontitis, information on the composition of their indigenous oral microbiota is limited. The aim of the current study was to characterize mouse oral bacterial flora by applying extensive parallel pyrosequencing using the latest model pyrosequencer, a Roche/454 Genome Sequencer FLX Titanium. In addition, the effect of Toll-like receptor (TLR) 2 deficiency on oral microbiota was evaluated.ResultsEight oral bacterial communities of wild-type (n = 4) and TLR2 knock-out (n = 4) C57BL/6 mice were characterized by analyzing 80,046 reads of 16S rRNA genes obtained by pyrosequencing. Excluding the PCR primers, the average length of each sequencing product was 443 bp. The average species richness of the murine oral bacterial communities was estimated to be about 200, but the communities were dominated by only two main phyla and several species. Therefore, the bacterial communities were relatively simple. The bacterial composition of the murine oral microbiota was significantly different from that of humans, and the lack of TLR2 had a negligible effect on the murine oral microbiota.ConclusionPyrosequencing using the Roche/454 FLX Titanium successfully characterized mouse oral bacterial communities. The relatively simple oral bacterial communities of mice were not affected by TLR2 deficiency. These findings will provide a basis for future studies on the role of periodontal pathogens in the murine model of periodontitis.
Highlights
Mice have long served as an animal model for periodontitis, information on the composition of their indigenous oral microbiota is limited
This study presents the first report of a 16S rRNA-based survey of a microbial community using the Roche/454 Genome Sequencer (GS) FLX Titanium system with > 400 bp sequence reads
The Roche/454 GS FLX Titanium system produced data sets with a longer average length than those generated by earlier models
Summary
Mice have long served as an animal model for periodontitis, information on the composition of their indigenous oral microbiota is limited. Since periodontitis is caused by a dental biofilm consisting of a complex microbial community rather than a single pathogen, information on the composition of indigenous oral microbiota is important. It is generally accepted that microbial community analysis should be culture-independent and utilize molecular identification methods such as sequencing of 16S rRNA genes. The typical procedure for culture-independent dissection of a bacterial community's structure involves the isolation of whole bacterial community DNA, amplification of 16S rRNA genes, cloning into an Escherichia coli host, and sequencing of each cloned amplicon. Pyrosequencing allows over 100-fold higher throughput than the conventional Sanger sequencing method.
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