中国畜牧兽医2024，Vol.51Issue(7)：3128-3141,14.DOI:10.16431/j.cnki.1671-7236.2024.07.037
1株牛源D∶L3∶L5型多杀性巴氏杆菌的分离鉴定及病原特性评价
Isolation,Identification and Pathogen Characteristics Evaluation of a Strain of Pasteurella multocida D∶L3∶L5 from Cattle
摘要
Abstract
[Objective]The experiment was aimed to explore the distribution of capsule serotypes,drug resistance and virulence genes of P asteurella multocida from a large-scale dairy farm in Xining city,Qinghai province,in order to provide reference for the treatment and prevention of respiratory diseases caused by Pasteurella multocida in cattle.[Method]Bacteria were isolated from the lung sample of a dead cattle.And then,through the methods of colony observation,dyeing microscopy,biochemical identification,specific primer amplification,16S rRNA clone sequencing analysis,phylogenetic tree construction,capsular serum and lipopolysaccharide type,virulence gene detection,pathogenicity test and drug sensitivity test,the isolated Pasteurellamultocida was identified and its pathogen characteristics were evaluated.[Result]The colonies isolated from the lung samples were dewdrop-like with a slight depression in the gray middle,and no hemolysis ring appeared.Gram staining showed coccidia pink,Wright's staining showed blue coccidia with dense staining at both ends.Biochemical identification showed that the results of D-glucose,D-mannitol,D-mannose,tyrosine arylaminase,phosphatase,COURMARATE and ELLMAN reaction were positive.16S rRNA sequencing results showed that the strain was more than 95％similar to a variety of Pasteurella multocida,and the isolated strain was identified as Pasteurella multocida and named XN222.The results of capsule serum typing and lipopolysaccharide type showed that XN222 was classified as type D,and its lipopolysaccharide types were L3 and L5.The results of virulence gene amplification showed that XN222 carried 19 virulence genes,including ptfA,fimA,hsf-2,sodA,tbpA,sodC,and so on.The results of pathogenicity test in mice showed that injection of 0.2 mL 1.5 ×108 CFU/mL bacterial solution resulted in 100％(10/10)death of mice,indicating that the isolated strain XN222 had strong pathogenicity.The results of drug sensitivity test showed that XN222 was sensitive to 15 kinds of antimicrobial agents,such as ceftiofur,levofloxacin,enrofloxacin,marbofloxacin,tetracycline and penicillin,and was moderately sensitive to tylosin.[Conclusion]In this study,a highly pathogenic Pasteurella multocida type D strain was isolated.The discovery of this strain enriched the domestic bovine subtype of Pasteurella multocida,and provided biological materials for the prevention and control of bovine respiratory diseases caused by Pasteurella multocida,etiological investigation and development of Pasteurella multocida multivalent vaccine.关键词
D型多杀性巴氏杆菌/分离鉴定/病原特性Key words
Pasteurella multocida type D/isolation and identification/pathogen characteristics分类
农业科技引用本文复制引用
赵奕朵,牛宇辉,马婷,冯晓微,EGIDE,刘明超,李建喜,张康,王磊,路晓荣,张景艳,程佳,郭志廷,徐国伟,黄莉萍..1株牛源D∶L3∶L5型多杀性巴氏杆菌的分离鉴定及病原特性评价[J].中国畜牧兽医,2024,51(7):3128-3141,14.基金项目
国家现代农业产业技术体系(CARS-36) （CARS-36）
中国农业科学院科技创新工程协同项目(CAAS-XTCX2016011-01-09) （CAAS-XTCX2016011-01-09）
中国农业科学院科技创新工程中兽医与临床团队建设项目(CAAS-ASTIP-2015-ILHPS-06) （CAAS-ASTIP-2015-ILHPS-06）
自治区科技特派员农村科技创业行动项目(2022BKY007) （2022BKY007）
通辽市科技计划项目(TLCXYD202102-02) （TLCXYD202102-02）
