四川地区牦牛源肺炎克雷伯氏菌的分离鉴定及其耐药性分析

Isolation,Identification and Drug Resistance Analysis of Klebsiella pneumoniae Isolated from Yak in Sichuan

为了分析四川地区牦牛源肺炎克雷伯氏菌耐药性并确定其耐药基因的携带情况,本试验采集了四川地区不同养殖场患呼吸道疾病牦牛肺脏、咽拭子、鼻拭子共127份病料组织,分离得到了43株肺炎克雷伯氏菌,并采用微量肉汤稀释法结合PCR扩增法检测43株肺炎克雷伯氏菌的耐药性和耐药基因携带情况。结果表明:43株肺炎克雷伯氏菌对氨苄西林、阿莫西林、多西环素、磺胺间甲氧嘧啶4种药物耐药率较高,为65.12%~90.70%;对阿米卡星、氟苯尼考、多黏菌素B、大观霉素4种药物的耐药率在20.93%~48.84%之间;对头孢噻呋、恩诺沙星、环丙沙星3药物的耐药率在18.60%~25.58%之间;多数筛选分离得到的菌株均呈现出多重耐药性,耐9、10种药物的菌株最多,分别占分离菌株的23.3%和20.9%。通过使用PCR扩增法检测分离菌携带的耐药基因情况,结果表明:blaTEM、blaSHV、sul2、sul3、floR 5种耐药基因检出率较高,检出率在62.8%~69.8%之间;ant(3″)-Ⅰa、aph(3′)-Ⅱa、aac(6′)-Ⅰb、aacC2 4种耐药基因检出率较低,检出率在7.0%~20.9%;blaCTX-M、Mcr-1、qnrA、qnrS、rmtB 5种耐药基因未检出。综上所述,本试验中分离得到肺炎克雷伯氏菌具有较强的耐药性并携带有较多的耐药基因,为了更好的促进四川省牦牛养殖产业的发展,应注意避免滥用抗生素并通过药敏试验对症下药。

In order to analyze the drug resistance of Klebsiella pneumoniae isolated from yak in Sichuan area,43 Klebsiella pneumoniae were isolated from 127 samples of lung,pharynx and nose swabs of yaks suffering from respiratory diseases in different feeding farms in Sichuan province.Microdilution broth method and PCR method were used to detect drug resistance and resistance genes of 43 strains of Klebsiella pneumoniae.The results showed that 43 strains of Klebsiella pneumoniae were resistant to ampicillin,amoxicillin,doxycycline and sulfamonomethoxine 4 drugs with a high resistance rate of 65.12%to 90.70%;The resistance rate to amikacin,florfenicol,polymyxin B and spectinomycin were 20.93%to 48.84%:And that to cefotafur,enrofloxacin,ciprofloxacin were 18.60%and 25.58%.Most isolates were multidrug resistant and 9 and 10 drug resistant strains accounted for 23.3%and 20.9%,respectively.The detection rates of blaTEM,blaSHV,sul2,sul3 and floR genes were high,ranging from 62.8%to 69.8%;The detection rates of ant(3″)-Ⅰa、aph(3′)-Ⅱa,aac(6′)-Ⅰb,aacC2 gene were low,ranging from 7.0%to 20.9%,and blaCTX-M,Mcr-1,qnrA,qnrS,rmtB genes were not detected.In conclusion,Klebsiella pneumoniae isolates in this experiment had strong drug resistance and carried multiple drug resistance genes.And in order to better promote the development of yak breeding industry in Sichuan province,more attention should be paid to avoid the abuse of antibiotics through drug sensitivity test.