This study aims to determine the prevalence of Campylobacter and its antibiotic susceptibility at slaughterhouses and live poultry markets in the Mekong Delta, Vietnam. A total of 177 samples, including 112 fecal samples, 28 cecal samples, and 37 rectal swab samples were collected. Thirty-nine samples tested positive for Campylobacter spp. by culture method, accounting for 22%. Isolation rates of Campylobacter in rectal swab samples were highest at 29.7% (11/37), followed by cecal samples at 25% (7/28) and fecal dropping samples at 18.8% (21/112). Species identification was performed by multiplex PCR and the result showed that the prevalence of Campylobacter coli (12.4%) was higher than Campylobacter jejuni (3.4%). Antibiotic susceptibility test showed that all isolated Campylobacter were susceptible to imipenem and meropenem (100%). Most Campylobacter were resistant to tetracycline (96.4%), ciprofloxacin (92.9%), nalidixic acid (89.3%), and ampicillin (67.9%). This study highlights the high contamination of Campylobacter and its antibiotic resistance in poultry in the Mekong Delta, Vietnam.