摘要
为分离西藏牦牛源地衣芽孢杆菌并研究其生物学特性,为今后开发相关益生菌及其制剂奠定基础,采集西藏牦牛粪便,从粪便中分离培养、纯化病原菌,通过染色镜检、生化试验、16S rRNA PCR扩增、同源性分析、进化树构建等对分离株进行鉴定且对分离株进行耐药性试验。结果从样品中分离到3株地衣芽孢杆菌,革兰氏阳性杆菌,分别命名为XZ-1,XZ-2,XZ-3。3株分离株生化检测结果一致:可分解甘露醇、淀粉、阿拉伯糖,可还原硝酸盐,V-P试验呈阳性,不能水解明胶,不能产生红色吲哚。分离株16S rRNA PCR扩增产物电泳结果与预期结果一致,长度为1 500 bp左右。3株分离株之间同源性为96.0%~99.9%,与地衣芽孢杆菌分离株的同源性为98.3%~100%;3株分离株与地衣芽孢杆菌处于同一支,而与链球菌其他种属芽孢杆菌处于不同分支。分离株XZ-1对青霉素、氨苄西林耐药,对林可霉素中度敏感,对头孢曲松、四环素、红霉素、恩诺沙星、氯霉素、环丙沙星、阿莫西林敏感;分离株XZ-2对青霉素、氨苄西林、四环素、林可霉素耐药,对氯霉素中度敏感,对头孢曲松、红霉素、恩诺沙星、环丙沙星、阿莫西林敏感;分离株XZ-3对青霉素耐药,对氨苄西林、对林可霉素中度敏感,对头孢曲松、四环素、红霉素、恩诺沙星、氯霉素、环丙沙星、阿莫西林敏感。
In order to isolate Bacillus licheniformis from Tibetan yaks and study its biological characteristics,lay a foundation for the development of relevant probiotics and their preparations in the future,Tibetan yak feces were collected,and pathogenic bacteria were isolated,cultured and purified from diseased materials.The isolated strains were identify through staining microscopy,biochemical tests,16S rRNA PCR amplification,homology analysis,and evolutionary tree construction.Drug resistance test was performed on the isolated strains.Three strains of Bacillus licheniformis and Gram-positive bacilli were isolated from the samples,which were named XZ-1,XZ-2 and XZ-3 respectively.The biochemical test results of the three isolated strains are consistent,which can decompose mannitol,starch,arabinose,reduce nitrate,and V-P test is positive.They cannot hydrolyze gela⁃tin and produce red indole.The electrophoresis result of 16S rRNA PCR amplification product of the isolate strains was consistent with the expected result,and the length was about 1500 bp.The homology between the three isolated strains was 96.0%-99.9%,and that with Bacillus licheniformis was 98.3%~100%;The three isolates are in the same branch with Bacillus licheniformis,but in different branches with other species of Bacillus strep⁃tococcus.The isolated strain XZ-1 is resistant to penicillin and ampicillin,moderately sensitive to lincomycin,and sensitive to ceftriaxone,tetracy⁃cline,erythromycin,enrofloxacin,chloramphenicol,ciprofloxacin and amoxicillin;The isolated strain XZ-2 is resistant to penicillin,ampicillin,tetra⁃cycline and lincomycin,moderately sensitive to chloramphenicol,and sensitive to ceftriaxone,erythromycin,enrofloxacin,ciprofloxacin and amoxicil⁃lin;The isolated strain XZ-3 is resistant to penicillin,moderately sensitive to ampicillin and lincomycin,and sensitive to other drug bacteria.
作者
王冬经
WANG Dongjing(Institute of Animal husbandry and veterinary,Tibet Academy of Agricultural and Animal Husbandry Science/State Key Laboratory of Highland Bar-ley and Yak Germplasm Resources and Genetic Improvement,Tibet Lhasa 850000,China)
出处
《西藏农业科技》
2023年第3期70-74,共5页
Tibet Journal of Agricultural Sciences
基金
西藏自治区科技重大专项(XZ202101ZD002N-05-01)
西藏自治区重点研发及转化项目(XZ202201ZY0007N-01)。
关键词
牦牛
地衣芽孢杆菌
分离鉴定
Yak
Bacillus licheniformis
Isolation and identification
