[目的]探究西藏阿里地区某养殖场内牦牛发病和死亡病因，分析其病原菌生物学特性，为有效防治该病提供参考。[方法]无菌采集死亡牦牛病变组织，根据临床症状进行实验室诊断并分离培养病原菌。通过形态学观察、生化鉴定、16S rRNA PCR扩增并测序、kmt基因鉴定、荚膜血清分型、脂多糖分型对分离株进行鉴定，并探究分离株的药物敏感性和致病性。[结果]分离株在7％血清TSA培养基中长出灰白色、透明、露滴样菌落;革兰染色镜检可见两端浓染的革兰阴性短杆菌。生化鉴定结果显示，分离株在葡萄糖、鸟氨酸脱羧酶、氧化酶、山梨醇和甘露醇生化反应管中呈阳性。荚膜血清分型及脂多糖分型结果显示，分离株为多杀性巴氏杆菌荚膜血清A型脂多糖L3型。药敏试验结果显示，分离株对羧苄西林、氨苄西林、头孢唑林、庆大霉素、呋喃妥因、头孢呋辛、丁胺卡那、米诺环素和诺氟沙星均表现敏感。致病性试验结果显示，小鼠腹腔注射0。2 mL 1×108 CFU/mL分离株菌液后，死亡率为100％(5/5)，分离株具有较强致病性。[结论]本研究成功分离鉴定出1株血清A型脂多糖L3型多杀性巴氏杆菌，其对羧苄西林、氨苄西林、头孢唑林等药物敏感，致病性较强。研究结果为探究牦牛源多杀性巴氏杆菌的生物学信息和病原学特征提供了数据参考。
Isolation,Identification and Pathogenicity Analysis of a Strain of Pasteurella multocida A∶L3 from Yaks
[Objective]The objective of this study was to explore the etiology of disease and death of yaks in a breeding farm in Ali area,Tibet,and analyze the biological characteristics of the pathogenic bacteria,so as to provide reference for effectively preventing and controlling the disease.[Method]Tissues of dead yaks were collected aseptically,and laboratory diagnosis was carried out according to clinical symptoms,and pathogenic bacteria were isolated and cultured.The isolate was identified by morphological observation,biochemical identification,16S rRNA PCR amplification and sequencing,kmt gene identification,capsular serotyping and lipopolysaccharide typing.The drug sensitivity and pathogenicity of the isolate was investigated.[Result]The isolate grew grayish white,transparent and dewdroplet colonies in 7％serum TSA medium.Gram staining showed strong Gram-negative brevibacterium at both ends.The results of biochemical tests showed that the isolate was positive in glucose,ornithine decarboxylase,oxidase,sorbitol and mannitol biochemical reaction tubes.The results of capsular serotyping and lipopolysaccharide typing showed that the isolate was Pasteurella multocida capsular type A,and its lipopolysaccharide type was L3.Drug susceptibility test showed that the isolate was sensitive to carbenicillin,ampicillin,cefazolin,gentamicin,nitrofurantoin,cefuroxime,amicana,minocycline and norfloxacin.The results of pathogenicity test showed that the mortality rate of mice was 100％(5/5)after injecting 0.2 mL 1 × 108 CFU/mL of the isolate bacterial solution,and the isolate had strong pathogenicity.[Conclusion]In this study,a strain of serotype A lipopolysaccharide L3 Pasteurella multocida was successfully isolated and identified,which was sensitive to carbenicillin,ampicillin,cefazolin and other drugs,and was highly pathogenic.The results provided data reference for exploring the biological information and etiological characteristics of Pasteurella multocida from yaks.
Pasteurella multocidayakisolation and identificationpathogenicity
万方数据
维普
