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利用全基因组单核苷酸多态性分析鉴定日本猪急性丹毒的豕丹毒杆菌克隆谱系

Clonal Lineages of Erysipelothrix rhusiopathiae Responsible for Acute Swine Erysipelas in Japan Identified by Using Genome-Wide Single-Nucleotide Polymorphism Analysis.

作者信息

Ogawa Yohsuke, Shiraiwa Kazumasa, Ogura Yoshitoshi, Ooka Tadasuke, Nishikawa Sayaka, Eguchi Masahiro, Hayashi Tetsuya, Shimoji Yoshihiro

机构信息

National Institute of Animal Health, National Agriculture and Food Research Organization (NARO), Tsukuba, Ibaraki, Japan.

Department of Bacteriology, Faculty of Medicine Sciences, Kyushu University, Fukuoka, Japan.

出版信息

Appl Environ Microbiol. 2017 May 17;83(11). doi: 10.1128/AEM.00130-17. Print 2017 Jun 1.

Abstract

causes swine erysipelas, an important infectious disease in the swine industry. In Japan, the incidence of acute swine erysipelas due to serovar 1a has recently increased markedly. To study the genetic relatedness of the strains from the recent cases, we analyzed 34 serovar 1a swine isolates collected between 1990 and 2011 and further investigated the possible association of the live Koganei 65-0.15 vaccine strain (serovar 1a) with the increase in cases. Pulsed-field gel electrophoresis analysis revealed no marked variation among the isolates; however, sequencing analysis of a hypervariable region in the surface-protective antigen A gene () revealed that the strains isolated after 2007 exhibited the same genotype and could be differentiated from older strains. Phylogenetic analysis based on genome-wide single-nucleotide polymorphisms (SNPs) revealed that the Japanese strains examined were closely related, showing a relatively small number of SNPs among them. The strains were classified into four major lineages, with Koganei 65-0.15 (lineage III) being phylogenetically separated from the other three lineages. The strains isolated after 2007 and the two older strains constituted one major lineage (lineage IV) with a specific genotype (M203/I257-SpaA), while the recent isolates were further divided into two geographic groups. The remaining older isolates belonged to either lineage I, with the I203/L257-SpaA type, or lineage II, with the I203/I257-SpaA type. These results indicate that the recent increased incidence of acute swine erysipelas in Japan is associated with two sublineages of lineage IV, which have independently evolved in two different geographic regions. Using large-scale whole-genome sequence data from isolates from a wide range of hosts and geographic origins, a recent study clarified the existence of three distinct clades (clades 1, 2, and 3) that are found across multiple continents and host species, representing both livestock and wildlife, and an "intermediate" clade between clade 2 and the dominant clade 3 within the species. In this study, we found that the Japanese strains examined exhibited remarkably low levels of genetic diversity and confirmed that all of the Japanese and Chinese swine isolates examined in this study belong to clonal lineages within the intermediate clade. We report that genotyping of strains is a practical alternative to whole-genome sequencing analysis of the isolates from eastern Asian countries.

摘要

引起猪丹毒,这是养猪业中的一种重要传染病。在日本,由血清型1a引起的急性猪丹毒发病率最近显著上升。为了研究近期病例中菌株的遗传相关性,我们分析了1990年至2011年间收集的34株血清型1a猪分离株,并进一步调查了活的小金井65 - 0.15疫苗株(血清型1a)与病例增加之间的可能关联。脉冲场凝胶电泳分析显示分离株之间无明显变异;然而,表面保护性抗原A基因()高变区的测序分析表明,2007年后分离的菌株表现出相同的基因型,且可与较老的菌株区分开来。基于全基因组单核苷酸多态性(SNP)的系统发育分析表明,所检测的日本菌株密切相关,它们之间的SNP数量相对较少。这些菌株被分为四个主要谱系,小金井65 - 0.15(谱系III)在系统发育上与其他三个谱系分开。2007年后分离的菌株和两个较老的菌株构成一个具有特定基因型(M203/I257 - SpaA)的主要谱系(谱系IV),而近期分离株进一步分为两个地理组。其余较老的分离株属于具有I203/L257 - SpaA型的谱系I或具有I203/I257 - SpaA型的谱系II。这些结果表明,日本近期急性猪丹毒发病率的上升与谱系IV的两个亚谱系有关,它们在两个不同的地理区域独立进化。一项近期研究利用来自广泛宿主和地理来源的分离株的大规模全基因组序列数据,阐明了在多个大陆和宿主物种(包括家畜和野生动物)中发现的三个不同进化枝(进化枝1、2和3)的存在,以及该物种中进化枝2和占主导地位的进化枝3之间的一个“中间”进化枝。在本研究中,我们发现所检测的日本菌株遗传多样性水平极低,并证实本研究中检测的所有日本和中国猪分离株都属于中间进化枝内的克隆谱系。我们报告说,对该菌株进行基因分型是对东亚国家该分离株进行全基因组测序分析的一种实用替代方法。

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