To investigate the persistence risk ofCampylobacterspp. in poultry farms, and to study the virulence and antimicrobial resistance characteristics in the recovered strains, we collected 362 samples from breeding hen flocks, before and after disinfection. The virulence factors were investigated by targeting the genes;flaA,cadF,racR,virB11,pldA,dnaJ,cdtA,cdtB,cdtC,ciaB,wlaN,cgtB, andceuEby PCR. Antimicrobial susceptibility was tested and genes encoding antibiotic resistance were investigated by PCR and MAMA-PCR. Among the analyzed samples, 167 (46.13%) were positive forCampylobacter. They were detected in 38.7% (38/98) and 3% (3/98) of environment samples before and after disinfection, respectively, and in 126 (75.9%) out of 166 feces samples. In total, 78C. jejuniand 89C. coliisolates were identified and further studied. All isolates were resistant to macrolids, tetracycline, quinolones, and chloramphenicol. However, lower rates were observed for beta-lactams [ampicillin (62.87%), amoxicillin-clavulanic acid (47.3%)] and gentamicin (0.6%). Thetet(O)and thecmeBgenes were detected in 90% of resistant isolates. TheblaOXA-61gene and the specific mutations in the23S rRNAwere detected in 87% and 73.5% of isolates, respectively. The A2075G and the Thr-86-Ile mutations were detected in 85% and 73.5% of macrolide and quinolone-resistant isolates, respectively. All isolates carried theflaA, cadF, CiaB, cdtA, cdtB,andcdtCgenes. ThevirB11,pldA,andracRgenes were frequent in bothC. jejuni(89%, 89%, and 90%, respectively) andC. coli(89%, 84%, and 90%). Our findings highlight the high occurrence ofCampylobacterstrains exhibiting antimicrobial resistance with potential virulence traits in the avian environment. Thus, the improvement of biosecurity measures in poultry farms is essential to control bacterial infection persistence and to prevent the spread of virulent and resistant strains.