1株奶牛源溶血性曼氏杆菌的分离鉴定与生物学特性研究  

Isolation,identification and biological characteristics of a strain of Mannheimia haemolytica from dairy cattle

作　　者：陈昌胜 王丽扬 杨跃飞[1,2] 王彦红 张信军[1,2] 成大荣 CHEN Changsheng;WANG Liyang;YANG Yuefei;WANG Yanhong;ZHANG Xinjun;CHENG Darong(College of Veterinary Medicine,Yangzhou University,Yangzhou 225009,China;Jiangsu Co-innovation Center for Prevention and Control of Important Animal Infectious Diseases and Zoonoses,Yangzhou University,Yangzhou 225009,China)

机构地区：[1]扬州大学兽医学院,江苏扬州225009 [2]扬州大学江苏高校动物重要疫病与人兽共患病防控协同创新中心,江苏扬州225009

出　　处：《黑龙江畜牧兽医》2022年第16期76-81,136,共7页Heilongjiang Animal Science And veterinary Medicine

基　　金：江苏高校优势学科建设工程资助项目(PAPD);扬州大学大学生创新创业训练计划基金项目(x20210704)。

摘　　要：为了对送检奶牛肺脏组织进行病原菌的分离鉴定并研究分离病原菌的生物学特性,试验采用细菌的分离纯化、革兰氏染色镜检、理化试验及16S rDNA序列分析对分离菌进行鉴定,并检测其血清型、多位点序列分型(MLST)、毒力基因、致病性和耐药性。结果表明:该菌株能发酵葡萄糖、乳糖和麦芽糖等,不发酵甘露糖;革兰氏染色镜检显示该菌呈红色的球杆状,为革兰氏阴性菌;16S rDNA序列分析结果显示该菌株为溶血性曼氏杆菌,命名为202102NF,为血清型1型;202102NF的7个管家基因adk、aroE、deoD、gapDH、gnd、mdh和zwf序列号分别为1,2,1,2,1,2,1,序列型为ST1;202102NF携带LKTA、GCP、POMA、NANH、ADHE、HF等多种毒力基因;小鼠腹腔注射202102NF 18 h内死亡,镜检可见攻毒组的小鼠肝脏和肾脏发生肿大、颜色加深,肺部出现出血斑,肠道有大量炎性渗出物;202102NF对诺氟沙星中介,对多黏菌素B、头孢噻肟、左氧氟沙星等17种抗菌药物均敏感。说明202102NF为血清型1型的溶血性曼氏杆菌,其序列型为ST1,携带多种毒力基因,具有较强的毒力,并对多种抗生素敏感。In order to isolate and identify pathogenic bacteria in the lung tissue of cows sent for inspection,and to study the biological characteristics of the isolated bacteria,the experiment used bacterial isolation and purification,Gram staining microscopy,physical and chemical tests,and 16 S rDNA sequence analysis to identify the isolated bacteria.The serotype,multilocus sequence typing(MLST),virulence genes,pathogenicity and drug resistance were detected.The results showed that the strain could ferment glucose,lactose and maltose,but not mannose.Gram staining microscopic examination showed that the bacteria were spherically rod-shaped and Gram-negative.The results of 16 S rDNA sequence analysis showed that the strain was Mannheimia haemolytica,named 202102 NF,which was serotype 1.The sequence numbers of the seven housekeeping genes adk,aroE,deoD,gapDH,gnd,mdh and zwf of 202102 NF were 1,2,1,2,1,2,1,respectively,and the sequence type was ST1.202102 NF carried LKTA,GCP,POMA,NANH,ADHE,HF and other virulence genes.Mice died within 18 h after 202102 NF intraperitoneal injection;microscopic examination showed that the livers and kidneys of the mice in the challenge group were enlarged and darkened;hemorrhage spots appeared in the lungs,and there were a large amount of inflammatory exudates in the intestines.202102 NF was intermediate to norfloxacin and sensitive to polymyxin B,cefotaxime,levofloxacin and other 17 antibiotics.The results suggested that 202102 NF was serotype 1 of Mannheimia haemolytica,and its sequence type was ST1,which carried a variety of virulence genes,with strong toxicity,and was sensitive to a variety of antibiotics.

关 键 词：溶血性曼氏杆菌 奶牛 16S rDNA基因序列 血清型 MLST分型 药敏试验 

分 类 号：S852.61[农业科学—基础兽医学]