一种蜱源性粪产碱杆菌内蒙古株的分离与鉴定  

Isolation and identification of a Alcaligenes faecalis IMH isolated from D. nuttalli in inner Mongolia

作　　者：贺海燕[1] 黄天鹏[1] 黄芳 吴咪 朝木丽格 格日勒图[1] HE Hai-yan;HUANG Tian-peng;HUANG Fang;WU Mi;CHAO Mulige;GERI Letu(College of Veterinary Medicine,Inner Mongolia Agricultural University,Hohhot,Inner Mongolia 010018,China)

机构地区：[1]内蒙古农业大学兽医学院

出　　处：《中国兽医学报》2018年第8期1533-1538,共6页Chinese Journal of Veterinary Science

基　　金：国家自然科学基金资助项目(31660709);内蒙古创新引导基金资助项目;边境地区媒介生物外来动物疫病检测溯源技术研究资助项目(2017YFD0501800)

摘　　要：从内蒙古呼伦贝尔地区自然放牧的羊群中采集蜱虫样本，分离出可疑细菌，并采取常规方法和分子生物学方法对其进行种型鉴定。采用高盐普通琼脂培养基从收集的蜱虫体内分离和纯化培养可疑细菌，并进行革兰染色；对纯化的可疑细菌进行生理生化、药物敏感试验及对盐的耐受性鉴定；PCR方法扩增和测序16SrRNA基因序列并应用Mega6．0生物软件绘制出该细菌系统发育进化树。结果显示，分离菌革兰染色呈革兰阴性短杆菌；溶血性试验结果呈现α-溶血；药物敏感试验对头孢类抗生素敏感，对青霉素、两性霉素B耐药；该菌具有运动性并可在含有10％的NaCl的培养基中生长；蔗糖、甘露醇、麦芽糖、乳糖、葡萄糖等糖分解试验、吲哚试验、硝酸盐（还原）试验、明胶液化试验等结果为阴性；脲酶试验、氧化型与发酵型试验、西蒙氏枸橼酸盐试验、氧化酶试验结果为阳性；根据16S rRNA基因绘制的系统发育树可看出，本试验分离株与粪产碱杆菌遗传距离近，处在相同分支。本试验成功分离出1株新的蜱源性粪产碱杆菌（Alcaligenes faecalis），重新命名为Alcaligenes faecalisIMH（简称为A．faecalis IMH），其16SrRNA基因重新注册于GenBank中，注册号为：LC213619．1。本试验为进一步研究该菌的致病性奠定基础。A suspected bacterium was isolated in Dermacentor nuttalli collected from natural grazing sheep in Hulunbuir,Inner Mongolia. Normal method and molecular biology were used for species identification. High salt nutrient agar medium was selected to isolate and purify suspect bacterium and Gram staining. Phenotypic identification of suspect bacterium was conducted by biochem- ical,antimicrobial susceptibility and salt tolerability test. Using PCR amplification and 16S rRNA gene sequences and application of Mega6.0 bio-software to plot the phylogenetic tree of the bacterium. The results showed the bacterium was Gram-negative, rod-shaped and displayed alpha hemolysis. The isolated bacterium was susceptible to cephalosporin and resistance to penicillin and amphotericin B. It showed motility and tolerance to 10 % NaC1. Saccharose, mannitol, maltose, lactose, glucose, indolis, nitrate（reduction）, gelatin test showed negative. Urea, oxidation type and fermentation test, simon citrate test,oxidase test results were positive. The phylogenetic tree analysis showed that isolated strains and Alcaligenes faecalis belonged to the same branch. A strain of Derrnacentor derived Alcaligenesfaecalis was isolated and renamed as Alcaligenesfaecalis IMH （simply A. faecalis IMH） in the study. Its 16S rRNA gene has been registered in GenBank（registration No. LC213619.1）. The study laid a foundation for pathogenicity study of the Alcaligenes faecalis.

关 键 词：草原革蜱 粪产碱杆菌 生物学特征 分子学鉴定 

分 类 号：S855.1[农业科学—临床兽医学] S884.6[农业科学—兽医学]