title: Molecular characterization and genotypic antimicrobial resistance analysis of Campylobacter jejuni and Campylobacter coli isolated from broiler flocks in northern Italy.
abstract: 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.
authors.affiliation: Department of Comparative Biomedicine and Food Science, University of Padua, viale dell'Università, 16-35020, Legnaro, PD, Italy.
authors.forename: M
authors.initials: M
authors.lastname: Giacomelli
authors.forename: C
authors.initials: C
authors.lastname: Andrighetto
authors.forename: F
authors.initials: F
authors.lastname: Rossi
authors.forename: A
authors.initials: A
authors.lastname: Lombardi
authors.forename: L
authors.initials: L
authors.lastname: Rizzotti
authors.forename: M
authors.initials: M
authors.lastname: Martini
authors.forename: A
authors.initials: A
authors.lastname: Piccirillo
mesh_terms: D000818:Animals; D000890:Anti-Infective Agents / Q000494:pharmacology; D015373:Bacterial Typing Techniques / Q000662:veterinary; D002169:Campylobacter Infections / Q000453:epidemiology / Q000382:microbiology / Q000662:veterinary*; D017000:Campylobacter coli / Q000187:drug effects / Q000235:genetics* / Q000302:isolation & purification; D016123:Campylobacter jejuni / Q000187:drug effects / Q000235:genetics* / Q000302:isolation & purification; D002645:Chickens / Q000382:microbiology*; D016000:Cluster Analysis; D004269:DNA, Bacterial / Q000737:chemistry / Q000235:genetics; D024881:Drug Resistance, Bacterial / Q000235:genetics; D005408:Flagellin / Q000235:genetics; D056426:Genetic Loci; D014644:Genetic Variation; D005838:Genotype; D007558:Italy / Q000453:epidemiology; D018942:Macrolides / Q000494:pharmacology; D011201:Poultry Diseases / Q000453:epidemiology / Q000382:microbiology*; D019105:Random Amplified Polymorphic DNA Technique / Q000662:veterinary; D017422:Sequence Analysis, DNA / Q000662:veterinary
chemical_list: D000890:Anti-Infective Agents; D004269:DNA, Bacterial; D018942:Macrolides; C064192:flaB1 protein, bacteria; D005408:Flagellin; C067138:flaA protein, bacteria