1株奶牛源松鼠葡萄球菌的分离鉴定及耐药性分析

doi:10.16431/j.cnki.1671-7236.2021.04.037

摘要/Abstract

摘要： 为确定奶牛垫料中是否含有病原菌并深入了解垫料中优势生长菌的类型、耐药性及致病性等情况，本试验进行了细菌分离培养、革兰染色镜检、生化试验鉴定、16S rDNA序列分析及同源性比对、药敏试验、小鼠致病性试验。结果显示，分离菌在普通营养琼脂平板上形成圆形、表面光滑的白色菌落，血琼脂平板上形成黏稠、较大的白色菌落。革兰染色镜检结果显示，分离菌为革兰阳性，球型，呈葡萄串状排列，或单个散落。生化试验结果显示，分离菌对木糖、葡萄糖、麦芽糖、果糖、乳糖、硝酸盐还原等呈阳性反应，而蜜二糖、木糖醇、山梨醇、尿素、V-P反应等呈阴性反应。16S rDNA序列分析结果显示，扩增的16S rDNA序列长度为1 298 bp，与松鼠葡萄球菌的核苷酸同源性达99.85%~100%；系统进化树结果显示，分离菌与松鼠葡萄球菌处于同一分支。动物试验结果表明，分离菌株对试验小鼠有较强的致病性，以0.2 mL/只（7.9×10⁸ CFU/mL）菌液的剂量接种小鼠，在48 h内死亡率为60%（3/5）。药敏特性分析结果显示，分离菌株对复方新诺明、氨苄西林等7种药物敏感，对克林霉素、头孢噻肟和头孢呋辛中度敏感，对青霉素、红霉素和林可霉素耐药。本研究为奶牛源松鼠葡萄球菌的分离鉴定及防控提供了参考依据。

关键词: 松鼠葡萄球菌; 奶牛; 16S rDNA; 药敏特性分析

Abstract: In order to determine whether there were pathogenic bacteria in cow bedding and to further understand the types,drug resistance and pathogenicity of dominant growth bacteria in bedding,bacterial isolation and culture,Gram staining microscopy,biochemical test identification,16S rDNA sequence analysis and homology comparison,drug sensitivity test,pathogenicity test in mice were carried out.The results showed that the isolated bacteria formed round and smooth white colonies on the nutrient agar medium,while thick and large white colonies were formed on the blood nutrient agar medium.The results of Gam staining microscopy showed that the isolate was Gram-positive,spherical,arranged in clusters of grapes or scattered individually.The results of biochemical tests showed that the reaction of xylose,glucose,maltose,fructose,lactose and nitrate were positive,while melibiose,xylitol,sorbitol,urea and V-P test were negative.The 16S rDNA sequence analysis results showed that the nucleotide sequence of 16S rDNA was 1 298 bp,99.85% to 100% homology with Staphylococcus sciuri.The phylogenetic tree showed that the isolate was clustered Staphylococcus sciuri.The animal experiments showed that the isolated bacterium had strong pathagenicity to mice,the mortality was 60% when inoculated with 0.2 mL 7.9×10⁸ CFU/mL isolated strain in 48 h.Drug resistance analysis showed that the isolate was sensitive to 7 drugs,such as sulfamethoxazole,ampicillin,moderately sensitive to clindamycin,cefotaxime and cefuroxime,resistant to penicillin,erythromycin and lincomycin.This study could provide a reference for identification and prevention of Staphylococcus sciuri.

Key words: Staphylococcus sciuri; dairy cow; 16S rDNA; drug sensitivity analysis

中图分类号:

S852.61

田秋丰, 张红, 尹珺伊, 刘秋瑾, 张军, 王岩, 秦平伟, 朱庆贺, 苗艳, 陈楠楠, 张淑芬, 丁昕颖, 马姗姗, 王振. 1株奶牛源松鼠葡萄球菌的分离鉴定及耐药性分析[J]. 中国畜牧兽医, 2021, 48(4): 1482-1488.

TIAN Qiufeng, ZHANG Hong, YIN Junyi, LIU Qiujin, ZHANG Jun, WANG Yan, QIN Pingwei, ZHU Qinghe, MIAO Yan, CHEN Nannan, ZHANG Shufen, DING Xinying, MA Shanshan, WANG Zhen. Isolation,Identification and Drug Resistance Analysis of a Strain of Staphylococcus sciuri from Dairy Cow[J]. China Animal Husbandry and Veterinary Medicine, 2021, 48(4): 1482-1488.

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