狐肺炎大肠杆菌毒力基因和耐药基因检测及药物敏感性分析
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摘要
为了解河北地区狐肺炎大肠杆菌毒力基因、耐药基因的分布及耐药性情况,本研究采用PCR技术对从狐肺炎病例分离的81株大肠杆菌的毒力基因和耐药基因进行检测,采用K-B法检测这些大肠杆菌对16种抗菌药物的敏感性,采用试管二倍稀释法检测中药对大肠杆菌的最小抑制浓度(MIC)和最小杀菌浓度(MBC),采用腹腔注射法检测81株大肠杆菌对小白鼠的致病性。结果显示:此次共分离到81株大肠杆菌,且对小白鼠具有较强致病性;毒力基因irp2、fyuA、Ler、eaeA检出率分别为50.6%、56.8%、9.9%、23.5%;耐药基因blaTEM、blaOXA1、sul1、sul2、blaTEM1、blaSHV-1、blaCTX-M、strA-strB、aadA1、blaPSE1、sul3、tetA、tetB、tetC、tetD、ermF检出率分别为84.0%、69.1%、43.2%、38.3%、28.4%、4.9%、24.7%、29.6%、16.1%、9.9%、11.1%、9.9%、18.5%、8.6%、13.6%、3.7%,未检出耐药基因ermB、ermC;81株大肠杆菌均对磺胺间甲氧嘧啶、林可霉素等耐药;诃子、乌梅、五味子对81株大肠杆菌具有较好的体外抑菌作用,抑菌圈直径在20 mm～36 mm,MIC和MBC在7.81 mg/mL～62.50 mg/mL。本研究调查了河北地区狐肺炎大肠杆菌的流行情况,对狐肺炎病的防治具有重要的指导意义,更具公共卫生学意义。
The objectives of this study were to investigate the distribution of virulence genes, resistance genes and drug resistance of the Escherichia coli isolated from fox pneumonia in Hebei. The virulence and resistance genes, the antimicrobial resistance, the minimum inhibitory concentration(MIC) and minimum bactericidal concentration(MBC) to Chinese herbal medicines and the pathogenicity of the E.coli were detected and determined by PCR, K-B method, the double-dilution method and artificial infection test in mice, respectively. The results showed that a total of 81 E.coli were isolated from fox pneumonia, which had strong pathogenicity to mice after intraperitoneal injection. The detection rates of virulence gene irp2, fyuA, Ler and eaeA were 50.6%, 56.8%, 9.9% and 23.5%, respectively. The detection rates of resistance gene blaTEM, blaOXA1, sul1, sul2,blaTEM1, blaSHV-1, blaCTX-M, strA-strB, aadA1, blaPSE1, sul3, tetA, tetB, tetC, tetD and ermF were 84.0%, 69.1%, 43.2%,38.3%, 28.4%, 4.9%, 24.7%, 29.6%, 16.1%, 9.9%, 11.1%, 9.9%, 18.5%, 8.6%, 13.6% and 3.7%, respectively. While, the resistance genes ermB, ermC were not detected. The 81 isolates were resistant to sulfamonomethoxine, lincomycin, etc.;Terminalia chebula, Prunus mume and Schisandra chinensis had better antibacterial activity against the 81 isolates with the diameters of the inhibition zone ranging from 20 mm to 36 mm. and the MIC and MBC ranged from 7.81 mg/mL to 62.50 mg/mL.The study investigated the prevalence of E.coli isolated from fox pneumonia in Hebei regions, and it had important guiding significance for the prevention and control of fox pneumonia.
The objectives of this study were to investigate the distribution of virulence genes, resistance genes and drug resistance of the Escherichia coli isolated from fox pneumonia in Hebei. The virulence and resistance genes, the antimicrobial resistance, the minimum inhibitory concentration(MIC) and minimum bactericidal concentration(MBC) to Chinese herbal medicines and the pathogenicity of the E.coli were detected and determined by PCR, K-B method, the double-dilution method and artificial infection test in mice, respectively. The results showed that a total of 81 E.coli were isolated from fox pneumonia, which had strong pathogenicity to mice after intraperitoneal injection. The detection rates of virulence gene irp2, fyuA, Ler and eaeA were 50.6%, 56.8%, 9.9% and 23.5%, respectively. The detection rates of resistance gene blaTEM, blaOXA1, sul1, sul2,blaTEM1, blaSHV-1, blaCTX-M, strA-strB, aadA1, blaPSE1, sul3, tetA, tetB, tetC, tetD and ermF were 84.0%, 69.1%, 43.2%,38.3%, 28.4%, 4.9%, 24.7%, 29.6%, 16.1%, 9.9%, 11.1%, 9.9%, 18.5%, 8.6%, 13.6% and 3.7%, respectively. While, the resistance genes ermB, ermC were not detected. The 81 isolates were resistant to sulfamonomethoxine, lincomycin, etc.;Terminalia chebula, Prunus mume and Schisandra chinensis had better antibacterial activity against the 81 isolates with the diameters of the inhibition zone ranging from 20 mm to 36 mm. and the MIC and MBC ranged from 7.81 mg/mL to 62.50 mg/mL.The study investigated the prevalence of E.coli isolated from fox pneumonia in Hebei regions, and it had important guiding significance for the prevention and control of fox pneumonia.
引文
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[9]赖婧,刘洋,汪宇,等.800株不同动物源大肠杆菌的耐药性监测[J].中国兽医杂志,2011,47(4):12-14.
[10]展天松,王彦红,许梦怡,等.肠致病性大肠杆菌的分离鉴定和耐药性分析[J].黑龙江畜牧兽医,2015,(2):67-69.
[11]王艳萍,董林,郭时金,等.禽源性大肠杆菌HPI毒力岛irp2基因PCR检测方法的建立与应用[J].中国兽医杂志,2017,53(1):86-89.
[12]向双云,周珍辉,李玉冰,等.8种中草药对大肠杆菌的体外抑菌试验[J].黑龙江畜牧兽医,2016,(5):167-169.
[13]李树明.50味中药对8种畜禽病原菌抑菌对比研究[J].中兽医学杂志,2000,(4):5-8.
[2]武瑞兵,高玉敏,王鹏翔,等.牛肉源大肠杆菌的耐药性检测及相关耐药基因分布[J].中国畜牧兽医,2015,42(2):452-458.
[3]Guerra B,Junker E,Schroeter A,et al.Phenotypic and genotypic characterization of antimicrobial resistance in German Escherichia coli isolates from cattle,swine and poultry[J].Int JInfect Dis,2014,21(3):489-492.
[4]陈祥,赵娟,高崧,等.我国部分地区猪源大肠杆菌LEE和HPI毒力岛相关基因的检测[J].中国人兽共患病学报,2006,22(1):33-35.
[5]庄林林.三重LAMP检测禽源沙门氏菌、大肠杆菌磺胺耐药基因(sul1、sul2、sul3)研究[D].扬州:扬州大学,2016.
[6]吕彪,宋志元,储君,等.20种中草药提取物对4种常见病原菌体外抑菌试验[J].中兽医学杂志,2016,(4):4-6.
[7]张召兴,肖丽荣,李巧玲,等.26味中药对狐源致病性大肠埃希菌地方株体外抑菌试验[J].动物医学进展,2018,39(2):127-130.
[8]代如意,李莉,殷中琼,等.5味中药对鸡白痢沙门氏菌的体外联合抑茵研究[J].西北农林科技大学学报(自然科学版),2015,43(2):34-37.
[9]赖婧,刘洋,汪宇,等.800株不同动物源大肠杆菌的耐药性监测[J].中国兽医杂志,2011,47(4):12-14.
[10]展天松,王彦红,许梦怡,等.肠致病性大肠杆菌的分离鉴定和耐药性分析[J].黑龙江畜牧兽医,2015,(2):67-69.
[11]王艳萍,董林,郭时金,等.禽源性大肠杆菌HPI毒力岛irp2基因PCR检测方法的建立与应用[J].中国兽医杂志,2017,53(1):86-89.
[12]向双云,周珍辉,李玉冰,等.8种中草药对大肠杆菌的体外抑菌试验[J].黑龙江畜牧兽医,2016,(5):167-169.
[13]李树明.50味中药对8种畜禽病原菌抑菌对比研究[J].中兽医学杂志,2000,(4):5-8.