Aims This study determined the risk factors and characteristics of vancomycin-resistant Enterococci (VRE) among individuals working with animals in Malaysia. Methods and Results Targeted cross-sectional studies accompanied with laboratory analysis for the identification and characterization of resistance and virulence genes and with genotype of VRE were performed. VRE were detected in 9.4% (95% CI: 6.4613.12) of the sampled populations. Enterococcus faecium, Enterococcus faecalis and Enterococcus gallinarum were isolated, and vanA was detected in 70% of the isolates. Enterococcus faecalis with vanB was obtained from one foreign poultry worker. At least one virulence gene was detected in >50% of Ent center dot faecium and Ent.faecalis isolates. The esp and gelE genes were common among Ent.faecium (58.3%) and Ent.faecalis (78%), respectively. The VRE species showed diverse RAPD profiles with some clustering of strains based on the individual's background. However, the risk factors found to be significantly associated with the prevalence of VRE were age (OR: 5.39, 95% CI: 1.9814.61) and previous hospitalization (OR: 4.06, 95% CI: 1.3312.35). Conclusion VRE species isolated from individuals in this study have high level of vancomycin resistance, were genetically diverse and possessed the virulence traits. Age of individuals and history of hospitalization rather than occupational background determined VRE colonization. Significance and Impact of the Study This study provides comprehensive findings on the epidemiological and molecular features of VRE among healthy individuals working with animals.