The purpose of this study was to determine the prevalence of thermotolerant Campylobacter spp. and antimicrobial resistance profiles isolated from in livestock feces in Isfahan, Iran. A total of 400 fecal of livestock samples were collected, and the presence of Campylobacter species was studied by culture and polymerase chain reaction-based assays and antimicrobial susceptibility test. A total of 28 Campylobacter isolates including 22 Campylobacter jejuni and 6 Campylobacter coli were recovered from feces of livestock. The prevalence rates of Campylobacter spp. were observed in this study in sheep (10%), goat (8%), cattle (5.3%), and camel (4%). The highest prevalence of Campylobacter spp. was found in the summer (10%) and the lowest was in winter (4%). Among the isolates from livestock, both C. jejuni and C. coli from fecal samples had the highest frequency of tetracycline (75.1%) and ciprofloxacin (57.1%) resistance. The results of this study showed a high prevalence of Campylobacter spp. in livestock feces in Isfahan, Iran. The presence of Campylobacter in livestock feces can contaminate the environmental and human food chain. Therefore, detection of Campylobacter spp. in livestock-originated samples is important to identify possible sources of infection and to have that a better understanding of the epidemiology of infection virulence genes of isolates is considered essential.