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鸡源高产淀粉酶丁酸梭菌的分离鉴定
引用本文:李圣杰,丁轲,姜芳芳,魏嘉敏,李旺,李元晓,孙二刚.鸡源高产淀粉酶丁酸梭菌的分离鉴定[J].中国畜牧兽医,2015,42(11):3073-3079.
作者姓名:李圣杰  丁轲  姜芳芳  魏嘉敏  李旺  李元晓  孙二刚
作者单位:1. 河南科技大学宏翔发酵饲料实验室, 洛阳 471003;2. 河南宏翔生物科技有限公司, 汝州 467500;3. 河南省动物疫病与公共卫生重点实验室, 洛阳 471003
基金项目:河南省科技厅重大科技攻关项目(131100110300);河南科技大学SRTP项目(2014259)
摘    要:为了获得产淀粉酶的丁酸梭菌,本研究从鸡小肠表面黏液中进行丁酸梭菌的分离与筛选,首先对样品进行80℃水浴10 min除去非芽孢菌后,然后接种到TSN培养基进行菌株的分离与筛选。对分离到的菌株进行菌落形态、显微形态初步观察,然后对疑似丁酸梭菌的菌株进行产酸能力和淀粉酶酶活检测,最后对既产酸又高产淀粉酶的菌株进行生理生化鉴定和16S rDNA序列分析。结果表明分离到一株革兰氏阳性厌氧菌C.B1,菌体呈短杆状,能形成圆形或椭圆形芽孢,生理生化结果也符合丁酸梭菌的基本特征,16S rDNA序列长度为1450 bp,与丁酸梭菌的同源性高达99%以上,因此确定该分离菌株为丁酸梭菌,为进一步开发新的微生态制剂奠定了基础。

关 键 词:丁酸梭菌  鸡源  淀粉酶  分离  鉴定  
收稿时间:2015-05-04

Isolation and Identification of Producing High-yielding Amylase Clostridium butyricum from Chicken
LI Sheng-jie,DING Ke,JIANG Fang-fang,WEI Jia-min,LI Wang,LI Yuan-xiao,SUN Er-gang.Isolation and Identification of Producing High-yielding Amylase Clostridium butyricum from Chicken[J].China Animal Husbandry & Veterinary Medicine,2015,42(11):3073-3079.
Authors:LI Sheng-jie  DING Ke  JIANG Fang-fang  WEI Jia-min  LI Wang  LI Yuan-xiao  SUN Er-gang
Institution:1. Hongxiang Biological Feed Laboratory of Henan University of Science and Technology, Luoyang 471003, China;2. Henan Hongxiang Bio-Sci & Tech Co., Ltd., Ruzhou 467500, China;3. Key Laboratory of Animal Disease and Public Health of Henan Province, Luoyang 471003, China
Abstract:In order to obtain a producing amylase Clostridium butyricum,the chicken small intestinal mucous was collected to isolate and screen Clostridium butyricum.First of all,the samples were heated at 80℃ for 10 min for removing most non-spore bacteria,then the samples were inoculated on TSN medium.The suspected Clostridium butyricum strains were screened by colony morphology and microscopic morphology observation.The producing acid ability and amylase activity of the isolates were successively tested,so the C.B1 strain,both producing acid and producing high-yielding amylase,was selected to be identified by physiological and biochemical test and 16S rDNA sequence analysis.The results showed that C.B1 strain had the characteristics of gram-positive anaerobe,rod-shaped and forming a circular or oval spore,16S rDNA sequence was 1450 bp.Homology comparison analysis indicated that it was most closest to Clostridium butyricum with 99% homology.So C.B1 strain was identified as Clostridium butyricum,this study would lay the foundation for developing new microecological preparations.
Keywords:Clostridium butyricum  chicken  amylase  isolation  identification  
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