Lai Liying, Dai Jiao, Tang Huanyu, Zhang Shouming, Wu Chunyan, Qiu Wancen, Lu Chengping, Yao Huochun, Fan Hongjie, Wu Zongfu
College of Veterinary Medicine, Nanjing Agricultural University, Nanjing 210095, China; Key Lab of Animal Bacteriology, Ministry of Agriculture, Nanjing 210095, China; OIE Reference Lab for Swine Streptococcosis, Nanjing 210095, China.
Realbio Genomics Institute, Shanghai 200050, China.
Vet Microbiol. 2017 Jun;205:26-33. doi: 10.1016/j.vetmic.2017.04.030. Epub 2017 May 1.
Streptococcus suis (SS), an important pathogen for pigs, is not only considered as a zoonotic agent for humans, but is also recognized as a major reservoir of antimicrobial resistance contributing to the spread of resistance genes to other pathogenic Streptococcus species. In addition to serotype 2 (SS2), serotype 9 (SS9) is another prevalent serotype isolated from diseased pigs. Although many SS strains have been sequenced, the complete genome of a non-SS2 virulent strain has been unavailable to date. Here, we report the complete genome of GZ0565, a virulent strain of SS9, isolated from a pig with meningitis. Comparative genomic analysis revealed five new putative virulence or antimicrobial resistance-associated genes in strain GZ0565 but not in SS2 virulent strains. These five genes encode a putative triacylglycerol lipase, a TipAS antibiotic-recognition domain protein, a putative TetR family transcriptional repressor, a protein containing a LPXTG domain and a G5 domain, and a type VII secretion system (T7SS) putative substrate (EsxA), respectively. Western blot analysis showed that strain GZ0565 can secrete EsxA. We generated an esxA deletion mutant and showed that EsxA contributes to SS virulence in a mouse infection model. Additionally, the antibiotic resistance gene vanZ was identified and expression of vanZ conferred resistance to teicoplanin and dalbavancin in Streptococcus agalactiae. We believe this is the first experimental demonstration of the existence of the T7SS putative substrate EsxA and its contribution to bacterial virulence in SS. Together, our results contribute to further understanding of the virulence and antimicrobial resistance characteristics of SS.
猪链球菌(SS)是猪的一种重要病原体,不仅被认为是一种人畜共患病原体,而且还被公认为是抗菌药物耐药性的主要储存库,它会导致耐药基因传播到其他致病性链球菌物种。除了2型(SS2)之外,9型(SS9)是从患病猪中分离出的另一种流行血清型。尽管已经对许多猪链球菌菌株进行了测序,但迄今为止尚未获得非SS2毒力菌株的完整基因组。在此,我们报告了从一头患有脑膜炎的猪中分离出的SS9毒力菌株GZ0565的完整基因组。比较基因组分析揭示了菌株GZ0565中有五个新的假定毒力或抗菌药物耐药性相关基因,而在SS2毒力菌株中没有。这五个基因分别编码一种假定的三酰甘油脂肪酶、一种TipAS抗生素识别域蛋白、一种假定的TetR家族转录阻遏物、一种含有LPXTG结构域和G5结构域的蛋白以及一种VII型分泌系统(T7SS)假定底物(EsxA)。蛋白质印迹分析表明菌株GZ0565可以分泌EsxA。我们构建了一个esxA缺失突变体,并表明EsxA在小鼠感染模型中对猪链球菌毒力有贡献。此外,还鉴定出了抗生素抗性基因vanZ,并且vanZ的表达赋予无乳链球菌对替考拉宁和达巴万星的抗性。我们相信这是首次通过实验证明T7SS假定底物EsxA的存在及其对猪链球菌细菌毒力的贡献。总之,我们的结果有助于进一步了解猪链球菌的毒力和抗菌药物耐药性特征。