副猪嗜血杆菌的分离鉴定

Isolation and Identification of Haemophilus parasuis

副猪嗜血杆菌(Hps)为猪呼吸道疾病综合征(PRDC)的重要病原之一。在对广西65个猪场281份病猪组织样品进行PRDC病原学调查的基础上,对PCR检测的Hps阳性样品进行了细菌分离,并进行了生化特性、药敏试验、16 S RNA基因片段序列分析和基因组DNA的PCR指纹图谱分析。结果显示,11份(3.91%)检测样品为Hps阳性,且均为混合感染;从南宁市四塘、桂林市永福、玉林市容县和钦州市浦北分离出5株Hps,分离菌株的生化鉴定结果均符合Hps生化特性,且对头孢噻呋和头孢菌素高度敏感;其中3株为血清5型,1株为血清12型,另1株未能定型;分离菌株16 S RNA基因片段之间及与GenBank其他一些代表菌株的核苷酸同源性在99%以上。

Haemophilus parasuis（Hps） as one of the important pathogens of porcine respiratory disease complex（PRDC） has already captured more attention in pig industry.On the basis of epidemiological investigation of PRDC,the isolation and identification of Hps were conducted on 281 tissue samples from 65 pig farms.The results showed that 11（3.91%） samples were positive for Hps,and all co-infected with Hps＋（other pathogens） styles.Total of five Hps strains were isolated from samples from Nanning,Guilin,Yulin and Qinzhou.Five isolates were sensitive to ceftiufur and cefoselis antibiotics,the biochemical characteristics of isolates accorded with those of previous report on Hps.The sequences of 16sRNA gene fragments of isolates have over 99% nucleotide similarity among them,and with representative sequences complied from the GenBank.The DNA fingerprint analysis showed that five Hps isolates can be classified into three serotype groups,among which three isolates belong to serotype 5,one isolate is serotype 12,and another is outside fifteen serotypes.The results above further confirmed that five isolates are Hps,and also provided useful information for prevention and control of Haemophilus parasuis disease in Guangxi.