The irrational use of antibiotics in humans and animals is highly related to the emergence and increase of antibiotic-resistant bacteria worldwide. A cross-sectional survey aimed at evaluating the current level of practices regarding antibiotic use in farm animals in Rwanda was carried out countrywide. Interviews were conducted on 229 farmers rearing different types of animals. The study has revealed that almost all respondent farmers could name at least one antibiotic used in farm animals and peni-streptomycin was named by most of them (95.6%). The use of antibiotics in farm animals was observed in the majority of respondents (97.4%). It was found that 44.4 and 26.5% of respondents reported that they used antibiotics for disease prevention and growth promotion, respectively. The use of non-prescribed antibiotics in animals was also reported by more than the half of respondent farmers (55.6%). The majority of farmers had a moderate level of practices regarding antibiotic use in farm animals (73.5%), very few had a high level (26%) and only one respondent had a low level. The high level of practices in regard to antibiotic use in animals was associated with the location of the farm, the type of reared animals, and the rearing system. The results of this study give an insight into antibiotics usage practices in farm animals in Rwanda. The generated information can guide sensitizations and promotions of the prudent use of antibiotics among farmers in order to limit the increase of antibiotic resistance in the country.