Department of Comparative Biomedicine and Food Science, University of Padua, viale dell'Università, 16-35020, Legnaro, PD, Italy.
Avian Pathol. 2012 Dec;41(6):579-88. doi: 10.1080/03079457.2012.734915.
Genetic variability and genotypic antimicrobial resistance (AMR) of Campylobacter jejuni and Campylobacter coli from commercial broiler farms were investigated in this study. Campylobacter isolates were genetically characterized by random amplified polymorphic DNA (RAPD)-polymerase chain reaction (PCR) and flaA-SVR and flaB-SVR sequence-based typing. Eight RAPD types were identified in C. jejuni and three in C. coli, while 16 fla profiles were detected among all isolates. Further, 13 flaA-SVR and 13 flaB-SVR alleles were identified. Both typing methods detected a high level of genetic diversity, but fla-SVR typing showed a higher discriminatory power. Indeed, Simpson's index of fla typing (D=0.920) was higher than that of RAPD typing (D=0.814). Moreover, the association of flaA-SVR and flaB-SVR sequence analysis showed a higher discriminatory power compared with the sequence analysis of single loci. Isolates were also analysed by the mismatch amplification mutation assay PCR test and the detection of cmeB gene to determine the occurrence of genetic determinants of AMR to macrolides and fluoroquinolones and multidrug resistance. The A2074C and A2075G mutations in the 23S rRNA gene, the C257T mutation in the gyrA gene, and the cmeB gene were higher in C. coli (19.0%, 67.0%, 100.0% and 100.0%, respectively) than in C. jejuni (0.0%, 3.1%, 48.3% and 48.3%, respectively). This study showed a high degree of genetic diversity and a high prevalence of genetic determinants of macrolide resistance, fluoroquinolone resistance and multidrug resistance among C. jejuni and C. coli isolates from Italian commercial broiler farms.
本研究调查了商业肉鸡养殖场空肠弯曲菌和结肠弯曲菌的遗传变异和基因型抗菌药物耐药性(AMR)。通过随机扩增多态性 DNA(RAPD)-聚合酶链反应(PCR)和 flaA-SVR 和 flaB-SVR 序列分型对弯曲菌分离株进行遗传特征分析。在空肠弯曲菌中鉴定出 8 种 RAPD 型,在结肠弯曲菌中鉴定出 3 种,在所有分离株中检测到 16 种 fla 图谱。此外,还鉴定出 13 种 flaA-SVR 和 13 种 flaB-SVR 等位基因。两种分型方法均检测到高水平的遗传多样性,但 fla-SVR 分型显示出更高的区分能力。实际上,fla 分型的辛普森指数(D=0.920)高于 RAPD 分型(D=0.814)。此外,flaA-SVR 和 flaB-SVR 序列分析的关联显示出比单基因座序列分析更高的区分能力。还通过错配扩增突变试验 PCR 试验和 cmeB 基因检测来分析分离株,以确定大环内酯类和氟喹诺酮类药物和多药耐药性的 AMR 遗传决定因素的发生情况。23S rRNA 基因中的 A2074C 和 A2075G 突变、gyrA 基因中的 C257T 突变和 cmeB 基因在结肠弯曲菌(分别为 19.0%、67.0%、100.0%和 100.0%)中的发生率高于空肠弯曲菌(分别为 0.0%、3.1%、48.3%和 48.3%)。本研究表明,意大利商业肉鸡养殖场空肠弯曲菌和结肠弯曲菌分离株具有高度的遗传多样性和大环内酯类耐药、氟喹诺酮类耐药和多药耐药遗传决定因素的高流行率。