山羊溶血性曼氏杆菌2型的分离鉴定  

Isolation and identification ofMannheimia haemolytica type 2 from goats

作　　者：刘晓波 毛首会 李鹏飞 杜国玉 闫俊成 吴锦艳[2] 刘萍[2] 何继军[2] 尚佑军[2] 曹小安[2] 王桂琴[1] LIU Xiao-bo;MAO Shou-hui;LI Peng-fei;DU Guo-yu;YAN Jun-cheng;WU Jin-yan;LIU Ping;HE Ji-jun;SHANG You-jun;CAO Xiao-an;WANG Gui-qin(School of Agriculture,Ningxia University,Yinchuan 750021,China;State Key Laboratory for Animal Disease Control and Prevention/Lanzhou Veterinary Research Institute,Chinese Academy of Agricultural Sciences,Lanzhou 730046,China;Shanxi Herds Dairy Products Group Co.,Ltd.,Baoji 721000,China)

机构地区：[1]宁夏大学农学院,宁夏银川750021 [2]中国农业科学院兰州兽医研究所动物疫病防控全国重点实验室,甘肃兰州730046 [3]陕西和氏乳业集团有限公司,陕西宝鸡721000

出　　处：《中国兽医科学》2023年第4期485-490,共6页Chinese Veterinary Science

基　　金：“十四五”国家重点研发计划项目(2022YFD1602203);宁夏回族自治区重点研发计划项目(2021BBF02036);国家现代农业产业技术体系项目(CARS-39-04B)。

摘　　要：为了确定某养殖场奶山羊细菌性感染导致死亡的具体病原种类,从送检的肺和肝组织病料中分离纯化细菌。通过生化培养特性观察、16S rRNA和毒力基因的PCR扩增、血清型鉴定、药敏试验和致病性试验来鉴定病原种类。结果显示,从送检的组织中分离出2株革兰阴性球杆菌;16S rRNA和毒力基因LKT测序结果表明,所分离的病原菌为溶血性曼氏杆菌,血清型鉴定均为溶血性曼氏杆菌A2型,故可确定分离株为具有致病性的溶血性曼氏杆菌2型。药敏试验结果显示,分离株对羧苄西林、庆大霉素敏感,对氨苄西林、卡那霉素等中度敏感,而对复方新诺明具有耐药性。小鼠致病性试验表明,接种分离株后小鼠于24 h内陆续死亡,并从剖检小鼠肺中也检测到溶血性曼氏杆菌,表明该菌有一定的致病力。结果表明,造成山羊死亡的主要病原是溶血性曼氏杆菌,这为溶血性曼氏杆菌病的临床用药提供了参考。In order to determine the specific pathogen of death dairy goats caused by bacterial infection,the bacteria were isolated from lung and liver tissue materials sent for testing.Pathogen species were identified by biochemical culture characterization,PCR amplification of 16S rRNA and virulence genes,serotype identification,susceptibility testing,and pathogenicity testing.The results showed that 2 strains of bacillus Gram-negative were isolated from the tissues sent for testing.The results of 16S rRNA and virulence gene LKT sequencing showed that the isolated pathogenic bacteria were Mannheimia haemolytica,and the serotypes were all M.haemolytica A2.So it could be determined that the isolated strain was pathogenic M.haemolytica type 2.The results of drug susceptibility test showed that the isolates were sensitive to carboxicillin and gentamicin,and were moderately sensitive to ampicillin and kanamycin,and were resistant to co-trimoxazole.The pathogenicity test of mice showed that the mice died successively in 24 h after inoculation of isolates,and M.haemolytica bacteria were also detected in the lungs of autopsy mice,indicating that the bacteria had certain pathogenicity.In conclusions:the main pathogen causing goat death was M.haemolytica.This provided a reference for the clinical medication practice of haemolytic mancellosis.

关 键 词：山羊 溶血性曼氏杆菌 分离鉴定 药敏试验 

分 类 号：S852.61[农业科学—基础兽医学]