山羊支原体山羊肺炎亚种野毒株的分离鉴定

Isolation and Identification of Mycoplasma capricolum subsp.capripneumoniae Wild Strain

为获得山羊传染性胸膜肺炎病原野毒株,采集陕西杨凌某羊场疑似患山羊传染性胸膜肺炎山羊的鼻分泌物,进行PCR检测,病原分离培养,菌落形态观察,生化培养试验鉴定,PCR鉴定,基因序列测定分析,动物回归试验。结果显示,PCR检测鼻液分泌物呈山羊支原体山羊肺炎亚种阳性;菌落呈现中心脐、边缘光滑、油煎蛋状样;能发酵葡萄糖、不水解精氨酸、不分解尿素、能还原四唑氮、膜斑试验阴性、可液化马血清、消化酪蛋白,对洋地黄皂苷敏感;基因测序结果显示与GenBank中已收录的山羊支原体山羊肺炎亚种基因同源性为99.8%;动物回归试验结果显示,该菌株对山羊造成了严重的胸膜肺炎,具有致病性。结果表明,成功分离出1株山羊支原体山羊肺炎亚种野毒株。

To obtain the pathogen of contagious caprine pleuropneumonia(CCPP),the nasal secretions of goats suspected contagious caprine pleuropneumonia in a caprine farm were collected from Yangling of Shaanxi province and diagnosed by PCR.The pathogenic bacteria were isolated and cultured,following colony morphology observation,biochemical test,PCR identification,gene sequence analysis,animal regression experiment.These results suggested the nasal secretions were positive with Mycoplasma capricolum subsp.capripneumoniae(Mccp)by PCR.The purified strain under light microscope showed a central umbilical cord with a smooth edge,fried egg-like and its biochemical characteristics with fermenting glucose,non-hydrolyzed arginine,non-decomposition urea,reducing triazole nitrogen,negative for membrane spot test,horse serum liquefied,casein digested,sensitive to digitonin.Sequence analysis indicated that the isolate was shared 99.8% homology with the Mccp in GenBank.Animal pathogenicity test showed that the isolate induced typical pleuropneumonia with strong pathogenicity.In conclusion,a wild strain of Mccp in goat was isolated and identified in this study.