盘羊体内绵羊肺炎支原体的分离和鉴定

Isolation and identification of Mycoplasma ovipneumoniae in Argali

为查明某动物园的盘羊发生呼吸道疾病的原因,本研究在发病的盘羊群中无菌采集28份鼻腔棉拭子和1份肺组织病料样品。采用支原体专用培养基从4份鼻腔棉拭子和1份肺组织病料样品中分离出5株支原体,连续3次克隆纯化后,通过形态观察、理化试验、PCR及基因特征鉴定,证明分离到的支原体均为绵羊肺炎支原体(MO)。将支原体分离株气管接种4只60日龄健康绵羊,10 d后2只绵羊陆续出现呼吸症状,其中1只伴有明显的体温变化,峰值达41.2℃,其余3只人工感染羊和对照组的羊体温变化不明显。35 d迫杀剖检,1只人工感染羊的肺脏前叶粉色实变明显,显微病理变化主要呈现间质增生性肺炎,其它羊肺脏病变不明显;经培养鉴定和PCR检测,从4只人工感染羊的肺脏样品中再次分离出MO。结果表明引起盘羊发病的病原为MO,本研究为野生盘羊支原体肺炎的快速诊断和防治提供参考依据。

To determine the cause of a respiratory diseases in argali, which was prevalent in the zoo of Shihezi of Xinjiang province, twenty eight nasal swabs and one lung specimen were collected from the argali flock infected the disease. Five of Mycoplasmas were isolated from 4 nasal swabs and 1 lung specimen with modified Hayflick Medium. All isolates were identified as Mycoplasma ovipneumoniae by germiculture, colonial morphology, physiological and biochemical characteristics, respectively. In addition, the 16S rRNA genes of the isolates were amplified and sequenced which the homology were 99.9% to 99.7% with reference strain Y98 available in the GenBank. Furthermore, four healthy lambs were artificially infected with the isolates, respectively, and the lambs developed the clinical symptoms within 35 days post artificial infection which were similar to the natural infected argali. The results indicate that M.ovipneumoniae is responsible for the pleuropneumonic disease in the flock.