牛源乳腺致病性大肠杆菌JL05全基因测序及毒力和耐药基因分析  

Whole genome sequencing reveals the distribution of resistance and virulence genes of mammary pathogenic Escherichia coli JL05 from bovine

作　　者：曲星霖 肖丹 吴健[2] 马晓媛[3] 白翠[3] 王妍[1] 王楠[3] QU Xing-lin;XIAO Dan;WU Jian;MA Xiao-yuan;BAI Cui;WANG Yan;WANG Nan(Agricultural College of Yanbian University,Yanij 133000,China;Jilin Academy of Agricultural Sciences,Changchun 130033,China;Animal Disease Control Center of Jilin Provincal,Changchun 130062,China)

机构地区：[1]延边大学农学院,吉林延吉133000 [2]吉林省农业科学院,吉林长春130033 [3]吉林省动物疫病预防控制中心,吉林长春130062

出　　处：《中国预防兽医学报》2020年第12期1220-1225,共6页Chinese Journal of Preventive Veterinary Medicine

基　　金：吉林省科技发展计划项目(20180623033TC)。

摘　　要：为研究一株从奶牛乳样中分离到的乳腺致病性大肠杆菌(MPEC)JL05的进化分群情况、耐药表型和耐药基因之间的相关性、毒力因子分布情况,本研究采用PCR法、微量肉汤稀释法和高通量测序法对其进化分群、16种抗菌药物的最小抑菌浓度(MIC)、毒力基因、耐药基因的检测和分析。PCR检测结果表明,MPEC JL05属于B2群。MIC检测结果表明,MPEC JL05除对磺胺异恶唑中介、对呋喃妥因敏感外,对β-内酰胺、头孢菌素、氨基糖苷、抗叶酸、喹诺酮、大环内酯、四环素、苯丙醇类等8类14种抗菌药物均耐药。全基因测序结果表明,MPEC JL05编码基因中存在MPEC特有的fec基因座;VFDB数据库分析表明MPEC JL05具有黏附、侵袭、转运、铁摄取、分泌系统、细菌毒素6类80个毒力基因,并发现了耶尔森菌强毒力岛(HPI);CARD数据库分析表明MPEC JL05菌株具有参与抗生素外排、抗生素失活、抗生素靶点改变、降低对抗生素的渗透性等4类耐药机制的53个耐药基因,且在质粒的耐药基因上下游发现了多个转座酶及整合酶基因。本研究揭示了MPEC JL05复杂的进化史,为更全面的认识MPEC基因组特征、致病力及耐药传播机制提供了参考。The aim of this study was to analyze the phylogenetic groups,virulence factor distribution,and the correlation between drug resistance phenotype and resistance genes of Escherichia coli JL05 isolated from milk samples of dairy cows.The PCR,microdilution method and high-throughput sequencing were used to investigate the phylogenetic groups,MIC of 16 antibacterial drugs,virulence genes and resistance genes.PCR results showed that E.coli JL05 belonged to group B2.MIC results showed that E.coli JL05 was sensitive to sulfamethoxazole and nitrofurantoin,but resistant to 14 of Class 8 antibacterial drugs,includingβ-lactam,cephalosporin,aminoglycoside,folic acid,quinolone,macrolide,tetracycline and phenylpropanol.The results of whole-genome sequencing showed that E.coli JL05 had fec locus which is specific to mammary pathogenic E.coli(MPEC).The analysis of the genome through VFDB database showed that E.coli JL05 had 80 virulence genes in six categories,including adhesion,invasion,transport,iron uptake,secretion system and bacterial toxin.In particular,Yersinia high pathogenicity island(HPI)was also detected.The analysis of the genome through CARD database showed that E.coli JL05 had 53 resistance genes involved in antibiotic efflux,antibiotic inactivation,change of antibiotic target and reduction of antibiotic permeability;and multiple trans-posase and integrase genes were found in the upstream and downstream of the plasmid resistance genes.This study revealed the complex evolutionary history of E.coli JL05,and provided a reference for a more comprehensive understanding of genomic characteristics,pathogenicity and drug resistance transmission mechanism of E.coli JL05.

关 键 词：大肠杆菌 进化分群 全基因组测序 耐药基因 毒力基因 

分 类 号：S852.61[农业科学—基础兽医学]