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内蒙古地区乳房炎奶样与环境中细菌的分离鉴定及耐药性分析
引用本文:钟华晨,王丽芳,郭晨阳,刘嘉琳,宋洁.内蒙古地区乳房炎奶样与环境中细菌的分离鉴定及耐药性分析[J].中国畜牧兽医,2023,50(2):817-826.
作者姓名:钟华晨  王丽芳  郭晨阳  刘嘉琳  宋洁
作者单位:内蒙古自治区农牧业科学院, 内蒙古自治区农牧业质量安全与检测研究所, 呼和浩特 010031
基金项目:国家自然科学基金项目(31860663);内蒙古自治区科技计划项目(2021GG0029);内蒙古自然科学基金项目(2021LHMS03014);内蒙古农牧业创新基金项目(2020CXJJM12);奶牛疾病绿色防控技术应用示范(2022TG16)
摘    要:【目的】研究内蒙古地区部分奶牛养殖场乳房炎生鲜乳样本及环境样本致病菌种类和耐药性,为乳房炎的防控提供临床用药指导和参考。【方法】采用平板划线法对内蒙古地区4个规模化养殖场468份(乳房炎生鲜乳样本199份,乳房炎奶牛饲养环境样本269份)样本进行细菌分离培养,采用形态学观察、革兰氏染色镜检以及16S rDNA测序对分离出的细菌进行鉴定,且对引发乳房炎的主要致病菌进行药敏性试验。【结果】高盐甘露醇培养基上呈现出小、白且偏黄色的菌落,经16S rDNA测序比对,与金黄色葡萄球菌高度相似的菌株共56株,分离率为11.97%;在伊红-美蓝培养基上呈现出黑紫色泛有金属光泽的菌落,经16S rDNA测序比对,与大肠杆菌高度相似的菌株共44株,分离率为9.40%。在199份生鲜乳样本中,金黄色葡萄球菌和大肠杆菌的分离率最高,分别为21.11%和17.09%;在269份环境样本中,金黄色葡萄球菌和大肠杆菌的分离率分别为5.20%和3.72%。药敏试验结果显示,金黄色葡萄球菌对青霉素、磺胺异噁唑、氨苄西林、阿莫西林/克拉维酸的耐药性相对较高,耐药率分别为90.63%、78.13%、75.00%、68.75%;对庆大霉素、环丙沙星、复方新诺明的敏感性较高。大肠杆菌对氨苄西林、头孢噻吩、磺胺异噁唑存在明显耐药性,耐药率分别为100%、94.29%、45.71%;对链霉素、卡那霉素、庆大霉素、多粘菌素E、美罗培南、阿莫西林/克拉维酸以及利福昔明表现出较高敏感性。【结论】内蒙古地区引发奶牛乳房炎的主要致病菌为金黄色葡萄球菌、大肠杆菌;金黄色葡萄球菌对青霉素、磺胺异噁唑、氨苄西林、阿莫西林/克拉维酸的耐药性相对较高;大肠杆菌对氨苄西林、头孢噻吩、磺胺异噁唑存在明显的耐药性。

关 键 词:奶牛乳房炎  致病菌  分离鉴定  耐药性  药敏试验  
收稿时间:2022-09-27

Isolation,Identification and Drug Resistance Analysis of Pathogenic Bacteria from Milk Samples and Environment of Mastitis in Inner Mongolia
ZHONG Huachen,WANG Lifang,GUO Chenyang,LIU Jialin,SONG Jie.Isolation,Identification and Drug Resistance Analysis of Pathogenic Bacteria from Milk Samples and Environment of Mastitis in Inner Mongolia[J].China Animal Husbandry & Veterinary Medicine,2023,50(2):817-826.
Authors:ZHONG Huachen  WANG Lifang  GUO Chenyang  LIU Jialin  SONG Jie
Institution:Institute of Quality Safety and Testing for Agricultural & Animal Husbandry of Inner Mongolia, Inner Mongolia Academy of Agricultural & Animal Husbandry Sciences, Hohhot 010031, China
Abstract:【Objective】 This study was aimed to investigate the species and drug resistance of pathogenic bacteria in raw milk samples and environmental samples of mastitis from some dairy farms in Inner Mongolia,so as to provide clinical medication guidance and reference for the prevention and control of mastitis.【Method】 Bacteria were isolated from 468 samples (199 samples of mastitis raw milk and 269 samples of mastitis cow breeding environment) from 4 large-scale farms in Inner Mongolia by plate streak method.The isolated bacteria were identified by morphological observation,Gram staining microscopy and 16S rDNA sequencing.Drug susceptibility testing was performed on the main pathogenic bacteria that cause mastitis.【Result】 There were small,white and yellow colonies on the high salt mannitol medium,56 strains were highly similar to Staphylococcus aureus by 16S rDNA sequencing and comparison,and the isolation rate was 11.97%.On eosin-methylene blue medium,there were black purple and metallic luster colonies.After 16S rDNA sequencing and comparison,44 strains were highly similar to Escherichia coli,with a separation rate of 9.40%.In 199 raw milk samples,the isolation rates of Staphylococcus aureus and Escherichia coli were the highest,it was 21.11% and 17.09%,respectively.In 269 environmental samples,the isolation rates of Staphylococcus aureus and Escherichia coli were 5.20% and 3.72%,respectively.The results of drug sensitivity test showed that Staphylococcus aureus was relatively resistant to penicillin,sulfamethoxazole,ampicillin,amoxicillin/clavulanic acid,and the drug resistance rates were 90.63%,78.13%,75.00% and 68.75%,respectively,and the sensitivity rate to gentamicin,ciprofloxacin and compound sulfamethoxazole was higher. Escherichia coli was significantly resistant to ampicillin,cefothiophene and sulfamethoxazole,and the drug resistance rates were 100%,94.29% and 45.71%,respectively,it was highly sensitive to streptomycin,kanamycin,gentamycin,polymyxin E,meropenem,amoxicillin/clavulanic acid and rifaximin.【Conclusion】 The main pathogenic bacteria causing dairy cow mastitis in Inner Mongolia were Staphylococcus aureus and Escherichia coli. Staphylococcus aureus had relatively high resistance to penicillin,sulfamethoxazole,ampicillin,amoxicillin/clavulanic acid, Escherichia coli had obvious drug resistance to ampicillin,cephalothiophene and sulfamethoxazole.
Keywords:diary cow mastitis  pathogenic bacteria  isolation and identification  drug resistance  drug sensitivity test  
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