Campylobacter species are a major cause of foodborne bacterial infections in both developed and developing countries worldwide.  Campylobacter jejuni is responsible for the majority of infections.  This study was conducted to identify virulence-associated genes in Campylobacter species isolated from livestock production systems in South Africa.  A total of 250 fecal samples consisting of cattle (n=50), chickens (n=50), goats (n=50), sheep (n=50) and pigs (n=50) were randomly collected from livestock in Eastern Cape and KwaZulu-Natal provinces of South Africa between April and October 2018.  The samples were analyzed for the presence of virulence genes in Campylobacter species using molecular PCR-based methods.  It was found that 77 and 23% of Campylobacter jejuni and Campylobacter coli respectively were isolated from all the livestock samples.  There were positive significant (P<0.05) correlations amongst all the virulence genes that were investigated.  Chi-square and Fisher’s exact tests were implemented to test for the effect of livestock species on the presence or absence of virulence genes.  The study demonstrated that most of livestock species can potentially cause zoonotic infections and food poisoning due to the high prevalence of Campylobacter.  The high prevalence of virulence genes highlights the significance of Campylobacter in livestock production systems in South Africa.  This requires the implementation of one-health approaches to reduce the impact of foodborne and zoonotic diseases for the welfare of human and animal health.