甘肃某地区奶牛运动场环境中耐药菌流行情况及相关耐药基因检测

doi:10.16431/j.cnki.1671-7236.2020.06.028

摘要/Abstract

摘要： 为了解甘肃某地区奶牛运动场环境中主要耐药性细菌的分布情况及耐药基因的携带情况，试验采用抗性平板筛选法分离、纯化耐药菌后再通过16S rRNA鉴定；采用PCR方法检测6类22种耐药基因携带情况。结果显示，两年两个奶牛运动场（A和B）分离出不同种属的耐药菌共665株，经16S rRNA PCR鉴定主要为大肠杆菌、屎肠球菌和普通变形杆菌，其中耐药性大肠杆菌数量最多（398株），A牛场耐药菌数两年相比增长28.50%，B牛场耐药菌数量持平；通过对样品进行10类药物筛选耐药菌结果显示，耐药率前3位的分别为糖肽类（11.13%，74/665）、四环素类和磺胺类（10.98%，73/665）及β-内酰胺类和氨基糖苷类（10.23%，68/665）。采用PCR方法对样品进行耐药基因检测发现，两年两个奶牛运动场的耐药基因携带情况基本相同，喹诺酮类耐药基因qnrB、qnrS的携带率分别为10.81%和83.78%，磺胺类耐药基因Sul1、Sul2和Sul3的携带率分别为100%、100%和56.76%，糖肽类耐药基因VanA、VanB和VanC的携带率分别为78.38%、100%和100%，四环素类耐药基因tetA、tetB、tetC、tetM、tetO、tetL的携带率分别为100%、100%、100%、100%、81.08%和75.68%，KPC、NDM、aac（6'）-Ⅰb-cr耐药基因的携带率分别为32.43%、29.73%及24.32%，未检出qnrC、tetK、tetW、VIM和OXA耐药基因。携带耐药基因与GenBank中登录参考株基因序列的同源性为98%~99%。甘肃地区奶牛运动场中耐药菌对常用抗菌药物普遍耐药且携带相应耐药基因，奶牛运动场中碳青霉烯类及氨基糖苷类耐药基因与上年结果相比数量增加，各相关单位人员应对其加强奶牛场中耐药菌的监测，临床治疗中应合理用药，以降低环境中耐药菌转移给人类的潜在风险。

关键词: 奶牛; 运动场环境; 耐药菌; 分离鉴定; 耐药基因

Abstract: In order to investigate the distribution of major resistant bacteria and the resistant genes around the environment of dairy cow pastureland in Gansu province,the resistant bacteria were isolated and purified by resistance plate screening method,and then identified by 16S rRNA in the study.The results showed that a total of 665 strains of different species of resistant bacteria were isolated from the two fields (farm A and farm B).The major strains were Escherichia coli,Enterococcus faecalis and Proteus genera by PCR identification of 16S rRNA.The number of resistant Escherichia coli was the largest (398 strains).The number of resistant bacteria in farm A increased by 28.50% over the past two years,while the number of resistant bacteria in farm B remained the same.The results of 10 kinds of resistant bacteria screening showed that the isolation rates of the first three categories were glycopeptide (11.13%,74/665),tetracycline and sulfonamide (10.98%,73/665),and beta-lactam and aminoglycoside (10.23%,68/665).PCR results showed that the resistant genes in dairy cow pastureland during two years period of time were almost similar.The prevalence rates of quinolone resistant genes qnrB and qnrS were 10.81% and 83.78%,respectively.The prevalence rates of sulfanilamide resistant genes Sul1,Sul2 and Sul3 were 100%,100% and 56.76%,respectively.The prevalence rates of glycopeptide resistance genes VanA,VanB and VanC were 78.38%,100% and 100%,respectively.The prevalence rates of tetracycline resistant genes tetA,tetB, tetC,tetM,tetO and tetL were 100%,100%,100%,100%,81.08% and 75.68%,respectively.The prevalence rates of KPC,NDM and aac(6')-Ⅰb -cr resistance genes were 32.43%,29.73% and 24.32%,respectively.However,qnrC,tetK,tetW,VIM and OXA resistance genes were not detected.The homology between resistance gene and the gene sequence of reference strain in GenBank was 98% to 99%.The resistant bacteria carrying corresponding resistance gene was very common in Gansu province.Especially,carbapenems and aminoglycoside resistance genes were detected in high number.Therefore,regular surveillance of drug-resistant bacteria was highly essential in order to minimize the risk caused by resistant bacteria to human.In addition,antibiotic drugs should be used wisely and in appropriate dose during clinical treatment.

Key words: dairy cow; pastureland environment; resistant bacteria; isolation and identification; resistence gene

中图分类号:

S852.61

张雪婧, 何卓琳, 侯晓, 蒲万霞. 甘肃某地区奶牛运动场环境中耐药菌流行情况及相关耐药基因检测[J]. 中国畜牧兽医, 2020, 47(6): 1882-1891.

ZHANG Xuejing, HE Zhuolin, HOU Xiao, PU Wanxia. Detection of Resistance and Resistance Genes of Dairy Cow Pastureland Environment in Gansu Province[J]. China Animal Husbandry and Veterinary Medicine, 2020, 47(6): 1882-1891.

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