Genotypic, Antimicrobial Resistance and Virulence Profiles of Thermophilic Campylobacter Isolates in Broilers

Bu çalışmanın amacı, Adana ve Hatay illerindeki ticari broyler çiftliklerinde Campylobacter jejuni ve Campylobacter coli'nin prevalansını, antimikrobiyal direnç ve genetik belirleyicilerini araştırmaktır. İzolatlar arasındaki genetik çeşitliliğin değerlendirilmesi, flaA geninin DdeI restriksiyon endonükleaz enzimi ile kesilmesine dayalı RFLP-Polimeraz Zincir Reaksiyonu (PZR) ve flaA ve flaB-SVR genlerinin kısa değişken bölgelerinin (SVR'ler) dizi analizi ile belirlendi. Ayrıca, izolatların antimikrobiyal duyarlılıkları disk difüzyon yöntemi ile yapıldı ve tetrasiklin (tetO), ampisilin (blaOXA-61), aminoglikozid ve çoklu dirence aracılık eden efluks pompası (cmeB) direnç genlerinin varlığı ise PZR ile araştırıldı. Siprofloksasine dirence neden olan mekanizmalar PZR ve takiben DNA dizi analizi ile araştırıldı. İzolatlar arasında 10 virulans (flaA, virB11, racR, cadF, ciaB, dnaJ and pldA) ve toksin geninin (cdtA, cdtB, cdtC) varlığı PZR ile incelendi. İncelenen 220 kloakal sıvabtan 194'ü C. jejuni (%89) ve 24'ü C. coli (%11) olmak üzere 218 (%99.1) Campylobacter spp. izole edildi. Tüm izolatlar kloramfenikol, gentamisin ve eritromisin'e duyarlı iken; C. jejuni ve C. coli izolatlarının nalidiksik asit, siprofloksasin, ampisilin, tetrasiklin ve amoksisilin-klavulanik asite direnç oranları sırasıyla %86.6-100, %86.6-100, %45.9-45.8, %43.3-50 ve %2.6-0.0 olarak belirlendi. C. jejuni ve C. coli izolatlarında tetO, blaOXA-61, cmeB ve aph-3-1 genlerinin prevalansı sırasıyla %45.3-54.4, %36.1-75, %1.5-83.3 ve %0.5-0 tespit edildi. C. jejuni ve C. coli izolatlarında flaA, cdtA, cdtB, cdtC, racR, cadF, ciaB, dnaJ ve pldA genlerinin prevalansı ise sırasıyla %100-100, %95.4-100, %94.3-100, %89.7-54.2, %89.2-79.2, %92.8-100, %67-16.7, %85.6-75 ve %80-66.7 olarak belirlendi. virB11 geni Campylobacter izolatlarının hiçbirinde tespit edilmedi. Tüm flaA ve flaB -SVR allelleri aynı PCR-RFLP patternleri gösterdi. Bu çalışmanın sonuçları broyler sürülerinde Campylobacter spp. prevalansının, patojenik potansiyelinin, genetik çeşitliliğinin ve antimikrobiyal direnç oranlarının yüksek olduğunu göstermektedir. Bu durum antimikrobiyallere dirençli Campylobacter'lerin ortaya çıkışını ve yayılımını azaltmak için etkili kontol önlemlerinin acilen uygulanması gerektiğini vurgulamaktadır

Broilerlerden İzole Edilen Termofilik Campylabacter’lerin Genotipik, Antimikrobiyal Direnç ve Virulens Profilleri

The objective of this study was to investigate the prevalence, antimicrobial resistance and genetic determinants of resistance of Campylobacter jejuni and Campylobacter coli isolated from commercial broiler farms in Adana and Hatay provinces, Turkey. The assessment of the genetic diversity among the isolates was determined by flaA based RFLP-Polymerase Chain Reaction (PCR) with restriction enzyme DdeI and sequence analysis of short variable regions (SVRs) of flaA and flaB-SVR genes. Antimicrobial susceptibility of the isolates was performed by disk diffusion method and tetracycline (tetO), ampicillin (blaOXA-61), aminoglycoside and multidrug efflux pump (cmeB) resistance genes were investigated by PCR as well. The genes conferring resistance to ciprofloxacin was screened by PCR and following DNA sequencing. The presence of ten virulence (flaA, virB11, racR, cadF, ciaB, dnaJ and pldA) and toxin genes (cdtA, dtB, cdtC) among the isolates was also investigated using PCR. Out of 220 cloacal swabs, 218 (99.1%) Campylobacter spp. including C. jejuni (n=194; 89%) and C. coli (n=24; 11%) were isolated. While all the isolates were susceptible to chloramphenicol, gentamicin and erythromycin, resistance rates for C. jejuni and C. coli isolates to nalidixic acid, ciprofloxacin, ampicillin, tetracycline and amoxicillin-clavulanic acid were determined as 86.6-100%, 86.6-100%, 45.9-45.8%, 43.3-50% and 2.6-0.0%, respectively. The tetO, blaOXA-61, cmeB and aph-3-1 genes detected in C. jejuni and C. coli isolates were 45.3-54.2%, 36.1-75%, 1.5-83.3% and 0.5-0%, respectively. The prevalence of flaA, cdtA, cdtB, cdtC, racR, cadF, ciaB, dnaJ and pldA in C. jejuni and C. coli isolates was 100-100%, 95.4-100%, 94.3-100%, 89.7-54.2%, 89.2-79.2%, 92.8-100%, 67-16.7%, 85.6-75% and 80-66.7%, respectively. virB11 gene was not detected in any of Campylobacter isolates. All flaA and flaB-SVR alleles displayed same PCR-RFLP patterns. The results of this study revealed high prevalence, pathogenic potantial, genetic diversity and antimicrobial resistance of Campylobacter spp. in broiler flocks, which highlights urgent need for implementing effective contol measures to reduce emergence and spread of antimicrobial resistant Campylobacters

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