Simple SummaryNumerous studies have already reported the presence of different antibiotic-resistantEnterococcusspecies in food-producing animals, animal products and pet food. However, studies specifically evaluating antimicrobial resistance inEnterococcusspp. in ornamental animals and their food are scarce. Therefore, this study aimed to identifyEnterococcusspp. and their antibiotic-resistant patterns in ornamental animal feed, as this could lead to the spread of antimicrobial resistance to humans due to their close contact with these animals.AbstractEnterococcusis a bacterial genus that is strongly associated with nosocomial infections and has a high capacity to transfer and acquire resistance genes. In this study, the main objective was to evaluate the presence ofEnterococcusspecies in ornamental animal feed and characterize their antimicrobial resistance and virulence factors. Antimicrobial susceptibility was determined using 14 antimicrobial agents by the disk diffusion method, complemented by genotypic analysis to identifyEnterococcusspecies and the presence of 14 antimicrobial resistance and 10 virulence genes. From 57 samples of ornamental animal feed, 103Enterococcusisolates were recovered from 15 bird, 9 fish and 4 reptile feed samples.Enterococcusisolates were highly resistance to rifampicin (78%) and erythromycin (48%), and 48% of isolates were classified as multidrug-resistant.Enterococcus faecalis(36.7%) andE. faecium(31.7%) were the species most frequently identified. Most isolates carried the resistance genesermB (57%) andtetL (52%) and the virulence genes,cylL (52%) andesp(40%).Enterococcus gallinarumwas the species with the highest number of multidrug-resistant isolates (50%) and virulence genes (80%). These results highlight the high levels of antibiotic-resistantEnterococcusspp. present in ornamental animal feed and the growing interaction of these animals with humans as a public health concern.