Department of Veterinary Sciences, National Chiayi University, Chiayi, Taiwan, ROC.
Department of Microbiology, Immunology, and Biopharmaceuticals, National Chiayi University, Chiayi, Taiwan, ROC.
J Microbiol Immunol Infect. 2016 Dec;49(6):892-901. doi: 10.1016/j.jmii.2014.10.003. Epub 2014 Nov 11.
BACKGROUND/PURPOSE: Escherichia coli is a common pathogen to cause clinical and subclinical mastitis in cows. A total of 57 E. coli isolates from raw milk from cows were characterized genetically and biochemically.
Extended-spectrum β-lactamase (ESBL) genes, the mechanism for fluoroquinolone resistance, and variations in virulence genes and genomes of these E. coli isolates were investigated by the antimicrobial susceptibility test, simplex and multiplex polymerase chain reaction (PCR), and pulsed-field gel electrophoresis (PFGE).
All E. coli isolates were resistant to cloxacillin (100%) and to a lesser extent (50%) to tetracycline, neomycin, gentamycin, ampicillin, ceftriaxone, cefotaxime (CTX), and ceftazidime (CAZ). Nearly 70% of the isolates were resistant to at least two antimicrobials and 28.1% carried AmpA and AmpC genes simultaneously. The predominant bla gene was bla, followed by bla, bla, bla, and bla Among the six (10.5%) ESBL-producing E. coli carrying bla, bla, or bla, two isolates 31 of ST410 in the ST23 complex and 58 of ST167 in the ST10 complex were also resistant to ciprofloxacin, enrofloxacin, and levofloxacin, with mutations at codon 83 from serine to leucine and codon 87 from aspartic acid to asparagine in GyrA and at codon 80 from serine to isoleucine in ParC. These isolates were genetically diverse in pulsotype analysis, lacked toxin genes of human pathogenic E. coli and carried mostly the prevalent virulence genes fimH, papGII, and α-hemolysin.
Lacking virulence genes examined, genetic diverse E. coli isolates are unrelated to human pathogenic E. coli. Enhancing sanitation in milk processing and transportation is needed to eliminate multidrug-resistant (MDR), fluoroquinolone-resistant, and ESBL-producing E. coli isolates.
背景/目的:大肠杆菌是引起奶牛临床和亚临床乳腺炎的常见病原体。从奶牛生乳中分离出的 57 株大肠杆菌进行了遗传和生化特征分析。
通过药敏试验、单重和多重聚合酶链反应(PCR)以及脉冲场凝胶电泳(PFGE)检测这些大肠杆菌分离株的超广谱β-内酰胺酶(ESBL)基因、氟喹诺酮类耐药机制以及毒力基因和基因组的变异。
所有大肠杆菌分离株均对氯唑西林(100%)具有耐药性,对四环素、新霉素、庆大霉素、氨苄西林、头孢曲松、头孢噻肟(CTX)和头孢他啶(CAZ)的耐药性较弱(50%)。近 70%的分离株至少对两种抗菌药物耐药,28.1%同时携带 AmpA 和 AmpC 基因。主要 bla 基因是 blaCTX-M-15,其次是 blaTEM-1,blaSHV-12,blaCTX-M-3 和 blaOXA-1 。在 6 株(10.5%)产 ESBL 的大肠杆菌中,携带 blaCTX-M-15、blaTEM-1 或 blaSHV-12,其中 2 株为 ST23 复合群 ST410 型,5 株为 ST10 复合群 ST167 型,对环丙沙星、恩诺沙星和左氧氟沙星耐药,gyrA 中 83 位丝氨酸突变为亮氨酸,87 位天冬氨酸突变为天冬酰胺,parC 中 80 位丝氨酸突变为异亮氨酸。这些分离株在脉冲场凝胶电泳图谱分析中具有遗传多样性,缺乏人致病性大肠杆菌的毒素基因,携带大多数流行的毒力基因 fimH、papGII 和α-溶血素。
缺乏检测到的毒力基因,遗传上多样化的大肠杆菌与人类致病性大肠杆菌无关。需要加强牛奶加工和运输过程中的卫生,以消除多药耐药、氟喹诺酮类耐药和产 ESBL 的大肠杆菌。