Dewhirst Floyd E, Klein Erin A, Bennett Marie-Louise, Croft Julie M, Harris Stephen J, Marshall-Jones Zoe V
Department of Microbiology, The Forsyth Institute, 245 First street, Cambridge, MA 02142, USA; Department of Oral Medicine, Infection and Immunity, Harvard School of Dental Medicine, 188 Longwood Avenue, Boston, MA 02115, USA.
Department of Microbiology, The Forsyth Institute, 245 First street, Cambridge, MA 02142, USA.
Vet Microbiol. 2015 Feb 25;175(2-4):294-303. doi: 10.1016/j.vetmic.2014.11.019. Epub 2014 Dec 2.
The human oral microbiome is known to play a significant role in human health and disease. While less well studied, the feline oral microbiome is thought to play a similarly important role. To determine roles oral bacteria play in health and disease, one first has to be able to accurately identify bacterial species present. 16S rRNA gene sequence information is widely used for molecular identification of bacteria and is also useful for establishing the taxonomy of novel species. The objective of this research was to obtain full 16S rRNA gene reference sequences for feline oral bacteria, place the sequences in species-level phylotypes, and create a curated 16S rRNA based taxonomy for common feline oral bacteria. Clone libraries were produced using "universal" and phylum-selective PCR primers and DNA from pooled subgingival plaque from healthy and periodontally diseased cats. Bacteria in subgingival samples were also cultivated to obtain isolates. Full-length 16S rDNA sequences were determined for clones and isolates that represent 171 feline oral taxa. A provisional curated taxonomy was developed based on the position of each taxon in 16S rRNA phylogenetic trees. The feline oral microbiome curated taxonomy and 16S rRNA gene reference set will allow investigators to refer to precisely defined bacterial taxa. A provisional name such as "Propionibacterium sp. feline oral taxon FOT-327" is an anchor to which clone, strain or GenBank names or accession numbers can point. Future next-generation-sequencing studies of feline oral bacteria will be able to map reads to taxonomically curated full-length 16S rRNA gene sequences.
已知人类口腔微生物群在人类健康和疾病中发挥着重要作用。虽然对猫科动物口腔微生物群的研究较少,但人们认为它也起着类似的重要作用。为了确定口腔细菌在健康和疾病中所起的作用,首先必须能够准确识别存在的细菌种类。16S rRNA基因序列信息被广泛用于细菌的分子鉴定,也有助于建立新物种的分类学。本研究的目的是获得猫科动物口腔细菌的完整16S rRNA基因参考序列,将这些序列置于物种水平的系统发育型中,并为常见的猫科动物口腔细菌创建一个基于16S rRNA的精选分类学。使用“通用”和门选择性PCR引物以及来自健康和患有牙周病的猫的龈下菌斑样本的DNA构建克隆文库。还培养了龈下样本中的细菌以获得分离株。确定了代表171种猫科动物口腔分类群的克隆和分离株的全长16S rDNA序列。根据每个分类群在16S rRNA系统发育树中的位置,开发了一个临时的精选分类学。猫科动物口腔微生物群的精选分类学和16S rRNA基因参考集将使研究人员能够参考精确定义的细菌分类群。一个临时名称,如“丙酸杆菌属猫科动物口腔分类群FOT-327”,是克隆、菌株或GenBank名称或登录号可以指向的一个锚点。未来对猫科动物口腔细菌的下一代测序研究将能够将读数映射到经过分类学整理的全长16S rRNA基因序列上。