一株种鸡源奇异变形杆菌的分离鉴定及毒力基因分析  

Isolation,identification and virulence gene analysis of Proteus mirabilis from chicken

作　　者：刘盼盼 贺泂杰[1] 徐国伟 王学智[1] 王胜义[1] 张宝锁 孙研 崔东安[1] LIU Pan-pan;HE Jiong-jie;XU Guo-wei;WANG Xue-zhi;WANG Sheng-yi;ZHANG Bao-suo;SUN Yan;CUI Dong-an(Lanzhou Institute of Husbandry and Pharmaceutical Sciences,Chinese Academy of Agricultural Sciences Engineering Technology Research Center of Traditional Chinese Veterinary Medicine of Gansu Province,Lanzhou 7300501 China;Lanzhou Veterinary Institute,Chinese Academy of Agricultural Sciences,Lanzhou 730050,China;Ningxia Shunbao Agricultural limited liability company,Wuzhong,751600,China)

机构地区：[1]中国农业科学院,兰州畜牧与兽药研究所,甘肃省中兽药工程技术研究中心,甘肃兰州730050 [2]中国农业科学院兰州兽医研究所,甘肃兰州730046 [3]宁夏顺宝农业股份有限公司,宁夏吴忠751600

出　　处：《中国兽医科学》2021年第10期1287-1294,共8页Chinese Veterinary Science

基　　金：甘肃省重点研发计划项目(20YF3NA003);宁夏回族自治区重点研发计划项目(2019BEF02003);中央级公益性科研院所基本科研业务费专项(1610322020012)。

摘　　要：为探明宁夏地区某种鸡场雏鸡死亡率高的原因,本研究从7日龄死亡雏鸡肝脏分离到病原菌,通过革兰染色、生化试验和16S rRNA基因测序对其进行鉴定,并进行药敏试验、毒力基因检测及雏鸡攻毒试验研究。结果显示,分离株为菌体呈球状、棒状、短杆及长杆状等形状不一的革兰阴性菌,并具"迁徙生长"特征,生化鉴定结果与奇异变形杆菌符合率为99%,16S rRNA与奇异变形杆菌相似性达99%以上;分离菌株携带ureC、zapA、mrpA、ucaA、rsbA、pmfA、atfA、atfC等8种毒力基因,其中mrpA、aftA、pmfA、atfC、rsbA、ucaA存在部分氨基酸突变;分离菌株对阿莫西林/克拉维酸、新霉素敏感,对氟苯尼考中度敏感,对氨苄西林、恩诺沙星、四环素等13种抗菌药物耐药;分离菌株对雏鸡具有较强致病性。上述结果表明,本研究分离鉴定到一株种鸡源奇异变形杆菌,该分离株为多重耐药并携带8种毒力基因的致病性奇异变形杆菌,这为鸡奇异变形杆菌感染的防治及其分子流行病学研究奠定了基础。To determine the cause of the high mortality rate of chicken on a breeder farm located in Ningxia autonomous region,bacterial isolation and identification were conducted using the liver from chicken died at the age of 7-day-old by Gram staining,biochemical tests and 16 S r RNA gene sequencing.In addition,the antibiotics sensitivity test,virulence gene and chicken infection challenge were evaluated.The results showed that the isolates was Gram-negative bacteria of rod or sphere shapes with the characteristics of migratory growth,and biochemical identification showed 99% confidence in Proteus mirabilis.Meanwhile,the isolates was more than 99% homologous with Proteus mirabilis by 16 S r RNA sequencing and was named NXQY 22.Eight virulence genes(ure C,zap A,mrp A,uca A,rsb A,pmf A,atf A and atf C) were detected,and there were some amino acid mutations in mrp A,uca A,rsb A,pmf A,atf A and atf C by virulence gene sequences analysis.Furthermore,antibiotics sensitivity test revealed its high susceptible to amoxicillin/clavulanic acid and neomycin,moderately susceptible to florfenicol,and resistant to 13 other antibiotics.The challenge test demonstrated that this isolates had pathogenicity in chicken.In conclusion,in this study,a strain of Proteus mirabilis from breeding bird was isolated and identified,which is a multi-drug resistant pathogenic isolates and carries eight virulence genes.This research laid the foundation for the prevention and treatment of chicken Proteus mirabilis infection and its molecular epidemiology.

关 键 词：奇异变形杆菌 种鸡 分离鉴定 毒力基因 

分 类 号：S852.612[农业科学—基础兽医学]