“
“Background Mice do not develop periodontitis naturally, but experimental periodontitis can be induced by inoculating mice KU 57788 with a periodontal pathogen such as Porphyromonas gingivalis [1]. Experimentally induced periodontitis in mice has served as an animal model for human periodontitis. 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. Although the oral microbiota of several mouse strains have been characterized [2–4], these studies were based on cultivation. In addition, the isolates were identified by phenotypic characterization, including Gram staining, the catalase reaction, and commercial biochemical tests such as API strips. It is now generally accepted that microbial community Selleck AZD9291 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.
Recently, pyrosequencing, a new high-throughput DNA sequencing technique, has been introduced and employed in various microbiological disciplines. Pyrosequencing allows over 100-fold higher throughput than the conventional Sanger sequencing method. The higher throughput makes it possible to process large numbers of samples simultaneously and also makes it possible to detect rare species [5]. The utility MLN2238 of pyrosequencing in the characterization of microbial communities has been well documented for the Roche/454 Genome Sequencer (GS) 20 machine [5, 6] and the GS FLX system [7–9], which produce sequence reads of approximately 100 bp and 250 bp in length, respectively. At the end of 2008, a new pyrosequencer called GS FLX Titanium was developed; it generates fivefold more sequencing reads and an extended read length (~450 bp) compared to the PLEK2 GS FLX system. This latest model pyrosequencer
has been used for genome sequencing but has not been tested for culture-independent microbial community analysis based on 16S rRNA. The composition of indigenous microbiota seems to be the result of strong host selection and co-evolution [10]. The role of the immune system in the selection of indigenous microbiota has been demonstrated in several studies. The total cultivable oral microbiota of athymic nu/nu mice was dominated by Enterococcus faecalis, while that of nu/+ mice was dominated by Lactobacillus murinus [11]. In contrast, B-cell-deficiency had no apparent influence on the indigenous oral microbiota of mice [12]. Toll-like receptors (TLRs) are innate immune receptors that recognize microbial molecular patterns and mediate innate immune responses to microbes.
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