牦牛肺炎病原多杀性巴氏杆菌和殊异支原体的分子鉴定与分析

青海省刚察县某牦牛产业基地牦牛发生咳嗽,鼻孔有分泌物流出,呼吸困难等症状的疾病。为快速确诊发病牛的致病原因,及时防控和治疗,从发病和病死的牦牛中,采集鼻腔拭子和肺脏、胸腔积液,利用PCR方法进行分子鉴定与分析。结果显示:此牛场中引发肺炎的病原是A型多杀性巴氏杆菌和殊异支原体两种病原体混合感染,测序结果显示5个有效样品中鉴定的为完全相同的病原,P.multocida-GC1的Kmt基因与参考序列中国株CP031554/3/2/1同源性在99%以上,M.dispar-GC1与参考序列的16S rRNA基因同源性在97%以上。同时在遗传进化角度再次分析确认了鉴定的P.multocida与中国和俄罗斯的A型多杀性巴氏杆菌聚成一大支;鉴定的M.dispar与殊异支原体聚为一支。结果表明,青海刚察地区牦牛存在感染多杀性巴氏杆菌和殊异支原体的潜在风险,该研究为牦牛呼吸道疾病与肺炎的病原学调查和病原的流行病学调查提供了参考数据。

Molecular identification and analysis of Pasteurella multocida and Mycoplasma dispar in pathogen of yak Pneumonia

In a yak industrial base locating in Gangcha County of Qinghai Province, there were cough, nostrils secretion and difficulty breathing symptoms diseases occurring in yaks. In order to quickly diagnose the causes, and timely to prevent and treat the diseases, we collected the samples from the diseased yaks including nasal swabs, lung tissue and pleural effusion, PCR was performed for molecular identification and analsis. The results showed that the pathogens causing pneumonia in this farm were Pasteurella multocida and Mycoplasma dispar mixed infection, that five valid samples were identified as identical pathogens from the sequencing results. The Kmt gene of P. multocida -GC1 was over 99% homologous to the reference sequences Chinese strain CP031554/3/2/1;the 16S rRNA gene of M. dispar -GC1 was 97% homologous to the reference sequences. Meanwhile, it was reanalyzed from the perspective of genetic evolution, which confirming that P. multocida was clustered together with the Chinese and Russian P. multocida serotype A;M. dispar were grouped together. The results showed that the yaks existed the potential risk of mixed infection of P. multocida and M. dispar . This study provided reference data for the pathogens investigation and epidemiological studies for yaks pneumonia respiratory diseases.