›› 2015, Vol. 42 ›› Issue (11): 2902-2908.doi: 10.16431/j.cnki.1671-7236.2015.11.013

• 生物技术 • 上一篇    下一篇

牛链球菌的分离鉴定及生物信息学分析

祖菲, 段纲, 项勋, 李浩欣, 杨志远, 朱旗, 代飞燕, 常华   

  1. 云南农业大学动物科学技术学院, 昆明 650201
  • 收稿日期:2015-04-20 出版日期:2015-11-20 发布日期:2015-11-26
  • 通讯作者: 常华 E-mail:375845001@qq.com
  • 作者简介:祖菲(1993-),女,云南昆明人,硕士生,研究方向:兽医临床学,E-mail:2877155283@qq.com
  • 基金资助:
    云南省林业厅野生禽类重大动物疫病监测项目(K2400033);云南省高校兽医公共卫生重点实验室建设项目(A3007954)

Isolation,Identification and Bioinformatics Analysis of Streptococcus bovis

ZU Fei, DUAN Gang, XIANG Xun, LI Hao-xin, YANG Zhi-yuan, ZHU Qi, DAI Fei-yan, CHANG Hua   

  1. College of Animal Science and Technology, Yunnan Agricultural University, Kunming 650201, China
  • Received:2015-04-20 Online:2015-11-20 Published:2015-11-26

摘要: 本研究对某动物医院的一头病牛粪样进行细菌分离鉴定,PCR扩增细菌16S rDNA基因并测序,构建进化树,对该菌进行药敏试验分析及生长曲线的测定。结果显示,该菌在血琼脂、巧克力琼脂、LB琼脂等培养基上均能长出光滑凸起,边缘圆润,大小各异,颜色不同的菌落,而在MRS琼脂、高盐甘露醇琼脂上却不生长;生化鉴定结果显示,乳糖、麦芽糖、甘露醇等均为阴性,葡萄糖、西蒙氏柠檬酸盐、尿素均为阳性,上述结果初步鉴定此菌为牛链球菌;进化树的构建进一步证明了分离菌属于牛链球菌,其与牛链球菌RD09的同源性为99%;药敏试验结果显示,牛链球菌对强力霉素、卡那霉素极敏感,对红霉素高度敏感,对链霉素、新生霉素、复方新诺明、头孢曲松、庆大霉素均敏感,对氯霉素、头孢拉定、阿莫西林均耐药;从牛链球菌生长曲线可看出2~23 h是牛链球菌的高度繁殖期,23~28 h是稳定期,28 h以后开始衰退。本试验结果为进一步研究牛链球菌奠定基础。

关键词: 牛链球菌; 分离鉴定; 16S rDNA; 生物信息学

Abstract: In this study,the dung samples of a sick cattle from an animal hospital were detected by bacteria cultivation.The bacteria were cultured by different differential media,and analyzed by biochemical test,16S rDNA gene was amplified by PCR,then sequenced and constructed phylogenetic tree,drug sensitivity test and growth curve were carried out.The results showed that the bacteria could grow smooth bulge,rounded edges,different size,different colors of the bacterial colonies in blood agar,chocolate agar,LB agar and so on,but could not grow in MRS agar and high salt mannitol agar.The results of lactose,maltose,mannitol and so on were negative,the results of glucose,simmon's citrate and urea were positive,this bacteria were identified as Streptococcus bovis by these preliminary results,and confirmed by phylogenetic tree construction.This isolate exhibited 99% nucleotide sequence identity with Streptococcus bovis RD09.The drug sensitivity test result of Streptococcus bovis showed that it was extremely sensitive to doxycycline and kanamycin,highly sensitive to erythromycin,sensitive to streptomycin,novobiocin,cotrimoxazole,ceftriaxone and gentamycin,resistant to chloramphenicol,cephalosporins cefradine and amoxicillin.The growth curve of Streptococcus bovis indicated that a rapid growth phase of the bacteria was from 2 to 23 h,the stationary phase was from 23 to 28 h,the decline phase was after 28 h.These experimental data provided the prerequisite for further study of Streptococcus bovis.

Key words: Streptococcus bovis; isolation and identification; 16S rDNA; bioinformatics

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