犬源支气管败血波氏杆菌的分离鉴定和全基因组测序分析  

Isolation,Identification and Whole Genome Sequencing Analysis of a Canine Bordetella bronchiseptica Strain

作　　者：李雅卓 颜亚松 王昊 孔令聪[1,3] 马红霞 LI Yazhuo;YAN Yasong;WANG Hao;KONG Lingcong;MA Hongxia(College of Animal Medicine,Jilin Agricultural University,Changchun 130118,China;College of Life Sciences,Jilin Agricultural University,Changchun 130118,China;Jilin Province Key Laboratory of Research and Development and Creation of New Veterinary Drugs,Changchun 130118,China;Engineering Research Center of Ministry of Education for Bioreactor and Drug Development,Changchun 130118,China)

机构地区：[1]吉林农业大学动物医学院,吉林长春130118 [2]吉林农业大学生命科学学院,吉林长春130118 [3]吉林省新兽药研发与创制重点实验室,吉林长春130118 [4]生物反应器与药物开发教育部工程研究中心,吉林长春130118

出　　处：《中国兽医杂志》2024年第11期24-31,共8页Chinese Journal of Veterinary Medicine

基　　金：吉林省科技发展计划项目(YDZJ202203CGZH050,20210202033NC);中国工程院战略研究与咨询项目(JL2023-07)。

摘　　要：为明确致犬严重呼吸道症状的病原菌及其病原特征,本试验采集患有严重呼吸道症状病死犬肺脏脓包样本,通过病原菌的分离纯化和形态学观察、生化试验、16S rRNA基因测序鉴定、生长曲线绘制对分离菌株进行种属鉴定;通过微量肉汤稀释法检测分离菌株的药物敏感性;通过溶血试验和小鼠感染试验检测分离菌株的致病性;通过全基因组测序分析分离菌株基因环境。结果显示,本试验分离菌株为支气管败血波氏杆菌,命名为Z2。药敏试验结果显示,分离菌株Z2对头孢曲松、头孢喹肟、链霉素和替米考星耐药,对恩诺沙星、多西环素和氟苯尼考等11种药物敏感。该分离菌株具有一定的溶血活性,对小鼠的半数致死量(LD50)为5.01×10^(5)CFU/mL。该分离菌株全基因组大小为5257583 bp,含有4887个基因;仅携带3个耐药基因,介导其对磺胺类和氨基糖苷类药物耐药,但未检测到β-内酰胺类耐药基因;共有1169个致病性相关基因,其中粘附性毒力基因最多,包括细胞间粘附蛋白、百日咳粘附素、菌毛、鞭毛和被膜等毒力基因。结果表明,分离菌株Z2具有较高的耐药性和致病性。本试验结果为支气管败血波氏杆菌的致病性研究提供了一定的参考价值。To identify the pathogenic bacteria causing severe respiratory symptoms in dogs and understand its pathogenic characteristics,this study collected lung abscess samples from dogs that died of severe respiratory issues.The isolated bacteria species were identified through bacterial isolation and purification,morphological observation,biochemical tests,16S rRNA gene sequencing,and growth curve plotting.The drug susceptibility of the isolate was tested using the microdilution broth method,while its pathogenicity was assessed by hemolysis test and mouse infection assay.Whole genome sequencing was also performed to analyze the genomic environment of the isolate.The results showed that the isolated strain was identified as Bordetella bronchiseptica,named Z2.Drug sensitivity tests indicated that strain Z2 was resistant to ceftriaxone,cefquinome,streptomycin,and tilmicosin but was sensitive to 11 other drugs,including enrofloxacin,doxycycline,and florfenicol.The isolate exhibited hemolytic activity,with a median lethal dose(LD_(50))for mice of 5.01×10^(5) CFU/mL.The whole-genome size of the isolate was 5257583 bp,containing 4887 genes.Only three resistance genes were identified,mediating resistance to sulfonamides and aminoglycosides,but noβ-lactam resistance genes were detected.A total of 1169 virulence-related genes were identified,with adhesion virulence genes being the most abundant,including genes for intercellular adhesion protein,pertactin,fimbriae,flagella,and capsules.The results suggest that strain Z2 has high levels of both drug resistance and pathogenicity.This study provides valuable insights into the pathogenicity of Bordetella bronchiseptica.

关 键 词：支气管败血波氏杆菌 分离鉴定 耐药性分析 全基因组测序 

分 类 号：S852.61[农业科学—基础兽医学]