副猪嗜血杆菌的分离鉴定及药敏试验

Isolation,Identification and Antibiotic Susceptibility Test of Haemophilus parasuis

黑龙江省近几年频繁发生以多发性浆膜炎、关节炎、脑膜炎以及急性死亡为特征的传染病,为猪场造成严重经济损失,为了确诊是否有副猪嗜血杆菌感染,采集病死猪肝脏、脾脏和肺脏等病料,进行副猪嗜血杆菌的分离;对其理化特性进行鉴定,应用PCR方法对其16SrRNA基因扩增后进行克隆测序,将测序结果在GenBank上进行BLAST分析,把相近的基因序列应用DNAStar软件进行同源性和进化关系分析;用分离菌株进行药物敏感性试验,筛选敏感药物。结果分离出一种具有多形性的NAD依赖性菌株,经鉴定为副猪嗜血杆菌;16SrRNA基因进化分析结果表明,分离菌株与以往报道的副猪嗜血杆菌中国、日本和美国分离株属于同一亚群,而西班牙分离株属于单独的亚群;分离菌株对壮观霉素和头孢拉定高度敏感。

The purpose of this study is to determine if there is Glasser＇s disease in Heilongjiang,which provide a theory basis for the prevention and control of Glasser＇s disease.The dead piglets with polyserositis and arthritis were collected and the bacterium was isolated from the samples.Then the physical and chemical properties of isolated strain were identified,its 16S rRNA gene was amplified by PCR,and this DNA fragment was subsequently cloned and sequenced.The sequenced results were analyzed by BLAST in GenBank.The homology and evolution relations analysis of the similar sequences were done by DNAStar.The drug sensitivity testing was done by isolated strain.The results indicated that a pantomorphic NAD dependent strain was isolated,the results of the physical and chemical properties and PCR showed that was Haemophilus parasuis.The phyletic evolution analysis results of the isolated strain 16S rRNA gene showed that it was in same subgroup with the Haemophilus parasuis in China,Japan and American,however that was in single subgroup in Spain;drug sensitivity testing showed that it was hypersensitive to spectinomycin and cephradine.