一株猪丹毒杆菌的分离与鉴定-马光强

一株猪丹毒杆菌的分离与鉴定-马光强 一株猪丹毒杆菌的分离与鉴定 马光强1，刘丽娟1，岳筠3，李 毅3，文 明1,2，程振涛1,2*，周碧君1,2* (贵州大学动物科学学院, 贵州 贵阳 550025；2.贵州省动物疫病与兽医公共卫生重点实验室, 贵州 贵阳 550025；3.贵州省动物疫病预防控制中心，贵州 贵阳 550008） 摘要:应用细菌分离培养、形态学观察、生物化学鉴定和基因序列比较等技术对贵州省黔东南州某养殖场疑似猪丹毒病例进行病原分离鉴定，并通过动物回归实验验证病原致病力、药敏试验分析其耐药特性。结果显示该分离菌在血琼脂平板上生长有圆形、湿润、露滴样细小菌落，菌落周围产生浅绿色溶血环，革兰氏染色为阳性细小杆菌，该菌能够发酵葡萄糖、乳糖、果糖等糖类，硝酸盐还原阳性，硫化氢阳性，以上特征与猪丹毒杆菌生物学特性相同；分离菌株基因组大小约400bp和1500bp的16SrRNA的基因片段测序序列与GenBank数据库中Lm各地分离株株核苷酸同源性在93.8%-100%之间，其中与国内哈尔滨ZYL株，及国外日本Fujisawa株、瑞典B3950_81及ATCC19414参考株同源性为100%，这从分子水平上进一步证实分离菌株为猪丹毒杆菌，且各地分离菌株16SrRNA基因序列相对保守性，地域和物种的差异性不明显。动物回归试验显示，9.6×103CFU的剂量可致死小白鼠，表明分离株具有较强的致病力。药敏试验结果显示分离菌对大环类脂类药物最为敏感。研究结果为本病临床防控与治疗提供参考资料。 关键词:猪丹毒杆菌；分离鉴定；序列分析 Isolation and Identification of a Erysipelothrix rhusiopathiae in Swine MA Guang-qiang1,LIU Li-juan1,YUE Jun3, LI Yi3,WEN Ming1,2, CHENG Zhen-tao1,2*,ZHOU Bi-jun1,2* （1.College of Animal Science，Guizhou Uiversity，Guiyang 550025，China；2.Key laboratory of animal disease and veterinary public health in guizhou province，Guiyang 550025，China；3.Animal Disease Prevention and Control Center in Guizhou Province，Guiyang， Guizhou，550008， China） Abstract:A strain of swine Erysipelothrix rhusiopathiae, which were characterized by cultural character, shape observation, biochemical test, animal pathogenicity experiment and antibiotic sensitivity test were isolated from infected pig farms in Qiandongnan Prefecture of Guizhou province. The results showed that the isolated strain growth on blood agar plate with circular , moist small colony,like shape of dew drop, a light green hemolysis ring around a colony.It is a small gram positive bacillus and It can ferment glucose, lactose, fructose and some other sugars, nitrate reduction is positive, hydrogen sulfide is positive, the above features same as Erysipelothrix rhusiopathiae’s biological characteristics. Two specific 16S rRNA fragment about 400bp and 1500bp of E.rhusiopathiae was amplified, and the nucleotide homology between 93.8%-100% compared with part of local strains in GenBank, ZYL of Harbin, Fujisawa of Japan, B3950_81 of Sweden,Chiba9393 and ATCC19414 of Japan(100%) isolated in onset of animals is the highest, which is further confirms from the molecular level that a E.rhusiopathiae has been isolated, and Large genetic differences between the 16S rRNA gene of local strains isolated from different ways. The results can provide theoretical data for prevention and treatment of the 基金项目：贵州省农业攻关项目“畜禽源主要病原菌药物防控技术创新与示范”（黔科合NY[2014]3042号），贵州省科技创新人才团队项目（黔科合人才团队[2015]4016号）。 作者简介：马光强(1987-)，男，贵州石阡人，在读硕士研究生，主要从事动物疫病细菌学研究。 *通讯作者，Tel: 0851-88298078, Email: chengzhentao@sohu.com；as.bjzhou@gzu.edu.cn。