牛昏睡嗜血杆菌的分离鉴定及其应用研究

Isolation and Characterization of Haemophilus somnus from Cattle and Its Application Study

关键词:  昏睡嗜血杆菌  分离  培养  PCR鉴定  致病力

Key words:  Haemophilus somnus  isolation  culture  PCR  pathogenicity

摘要:

为明确国内流行牛昏睡嗜血杆菌病病原特性，将临床采集牛昏睡嗜血杆菌病诊断阳性病料划线接种于脑心浸液酵母浸提膏血琼脂固体培养基，经多代次最适固液培养基纯化分离培养，获得稳定传代的目标菌株，经8～12 h液体培养基培养，其菌数可达3×10⁹CFU/mL，对小鼠的最小致死剂量为4×10⁸ CFU，致病性较强。将培养菌液制备成灭活疫苗免疫小鼠后，能够针对流行的昏睡嗜血杆菌致病菌株产生80%以上免疫保护效果，且对小鼠安全性良好。该菌株的成功分离对牛昏睡嗜血杆菌病疫苗的研制具有促进作用。

Abstract:

In order to clarify the pathogen characteristics of the Haemophilus somnus disease in China, the clinical samples with positive diagnosis of Haemophilus somnus were inoculated on the solid medium of Brain Heart Extraction Yeast Extraction Ointment Blood Agar. The target strains were purified and cultured on the most suitable solid-liquid medium for many generations. After 8~12 hours of liquid medium culture, the number of bacteria could reach 3×10⁹CFU/mL, and the minimum lethal dose for all mice was 4×10⁸CFU，which is highly pathogenic. After immunizing mice with inactivated vaccine prepared from the culture liquid, it can produce more than 80% immune protection effect against the epidemic pathogenic strain of Haemophilus somnus, and the safety is good. The successful isolation of this strain will play a positive effect on the development of a vaccine against bovine Haemophilus somnus disease.