首页|额敏县1株羊粪源金黄色葡萄球菌的分离鉴定及其耐药性与致病性研究

额敏县1株羊粪源金黄色葡萄球菌的分离鉴定及其耐药性与致病性研究

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[目的]对羔羊腹泻粪便样本中的金黄色葡萄球菌进行分离鉴定,了解其耐药性和致病性特征.[方法]从新疆维吾尔自治区塔城地区额敏县某养羊场采集腹泻羔羊肛拭子样本 4 份,使用高盐甘露醇培养基(MSA)选择性分离金黄色葡萄球菌;通过形态学观察和 16S rDNA序列分析对分离菌株进行种属鉴定;采用纸片扩散法分析分离菌株对 9 类 11 种抗菌药物的敏感性;利用PCR法检测分离菌株毒力基因携带情况;建立昆明系小白鼠腹腔感染模型,开展分离菌株的致病性试验.[结果]从 4 份羔羊腹泻样本中分离到 1 株疑似金黄色葡萄球菌,该菌株在MSA培养基上形成有光泽、中度隆起、金黄色或柠檬色的菌落;革兰染色镜检可见分离菌株为革兰阳性,呈葡萄串状排列;16S rDNA序列分析显示,分离菌株的序列与我国湖南省长沙市猪肺源金黄色葡萄球菌分离株序列的相似性为 100%;结合形态学观察和 16S rDNA序列比对结果,鉴定该菌株为金黄色葡萄球菌.分离菌株对青霉素、克林霉素、头孢噻吩、利福平和红霉素耐药,对庆大霉素、阿奇霉素、氯霉素、诺氟沙星、头孢西丁和复方新诺明敏感.在检测的 8 类 20 种毒力基因中,该菌株携带sea、seg、sei、eta、clfA和clfB基因.该菌株对小鼠的半数致死量(LD50)为 2.24×108 CFU/mL,死亡小鼠脏器病理变化明显.[结论]从额敏县某养羊场羔羊腹泻样本中分离鉴定到 1 株金黄色葡萄球菌,该菌株具有多重耐药表型,携带多种毒力基因,具有一定致病性.研究结果为该地区羊金黄色葡萄球菌病的合理用药提供了参考.
Isolation,Identification,Antimicrobial Resistance and Pathogenicity of a Staphylococcus aureus Strain from Feces of Sheep in Emin County,China
[Objective]The aims of the present study were to isolate and identify the Staphylococcus aureus(S.aureus)strain from fecal samples of lambs with diarrhoea,and to characterize the antimicrobial resistance and pathogenicity of the isolate.[Method]Four anal swab samples of diarrheic lambs were collected from a sheep farm in Emin County,Tacheng Prefecture,Xinjiang Uygur Autonomous Region,and S.aureus strain was selectively isolated with mannitol salt agar(MSA).The isolate was identified by morphological observation and 16S rDNA sequence analysis.The in vitro susceptibility of the isolate to 11 antimicrobial agents belonging to 9 categories was analyzed by paper diffusion method.The presence of virulence genes of the isolate was detected by PCR assay.A model of abdominal infection in Kunming mice was established to evaluate the pathogenicity of the isolate.[Result]One suspected S.aureus strain was isolated from the four fecal samples of diarrheic lambs.It formed glossy,moderately raised,golden or lemon coloured colonies on MSA.Gram staining and morphological observation revealed that the isolate was Gram-positive and arranged in the form of grape bunches.The 16S rDNA sequence of the isolate was 100%similar to that of a S.aureus isolate of porcineluny origin from Changsha City,Hunan Province,China.Morphological observation and 16S rDNA sequence analysis confirmed that the isolate was S.aureus.The isolate was resistant to penicillin,clindamycin,cefotiamine,rifampicin and erythromycin,and was sensitive to gentamicin,azithromycin,chloramphenicol,norfloxacin,cefoxitin and cotrimoxazole.Out of the 20 virulence genes belonging to 8 classes tested,the presence of sea,seg,sei,eta,clfA and clfB genes was observed.The LD50 of the isolate for Kunming mice was 2.24×108 CFU/mL,with observable pathological changes in the organs of the dead mice.[Conclusion]One S.aureus strain was isolated and identified from the fecal samples of diarrheic lambs in a sheep farm in Emin County,which exhibited a multi-drug resistant phenotype,harbored multiple virulence genes and had certain pathogenicity.The results of this study provide references for the rational use of medication for the diseases caused by S.aureus infection in sheep in this region.

sheepStaphylococcus aureusisolationidentificationantimicrobial resistancepathogenicity

苏政、于月通、朱婷婷、陀海欣、李静、吴静、刘鹏、彭健、齐萌

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新疆生产建设兵团第九师农业科学研究所(畜牧科学研究所),新疆 额敏 834601

塔里木大学动物科学与技术学院/新疆生产建设兵团塔里木动物疫病诊断与防控工程实验室,新疆 阿拉尔 843300

金黄色葡萄球菌 分离 鉴定 耐药性 致病性

新疆生产建设兵团科技创新人才计划新疆生产建设兵团第九师科技计划项目新疆生产建设兵团第九师科技计划项目

2021CB0622021JS0012022JS008

2024

畜牧与饲料科学
内蒙古自治区农牧业科学院

畜牧与饲料科学

CSTPCD
影响因子:0.332
ISSN:1672-5190
年,卷(期):2024.45(4)