Background The inappropriate use of antibiotics is a major issue in clinical practice in Greece with serious implications for public health and animal health. The purpose of the present study was to provide a first insight into the use of antibiotics by small animal practitioners in Greece and assess their compliance with general rules for the rational use of antibiotics. This is the first survey of its kind in Greece.Methods A questionnaire was designed to collect basic information on the use of antibiotics by pet veterinarians. The questionnaire was sent to a total of 70 veterinarians mainly operating in the region of Attica, a region that comprises almost 50% of the Greek population and where veterinarians are engaged solely in small animal practice. The questionnaire consisted of 37 closed questions dealing with various aspects on the use of antibiotics.Results The majority of respondents’ report cases where the pet owner initiated antibiotic treatment without veterinary prescription. Almost every clinician reported owner-compliance challenges. Regarding microbiological analysis, 73% of respondents prefer to adopt the empirical treatment while waiting the laboratory results or use antibiogram only when the treatment is unsuccessful. Eighty-eight per cent declared to use antimicrobials postoperatively in clean surgical procedures. Different types of antimicrobials and treatment durations than the ones proposed by guidelines on rational use of antibiotics are preferred for various organ systems e.g. in urinary and gastrointestinal infections. Conclusions Our findings suggest the need for guidelines on antibiotic use in small animal practice in Greece, and the deployment of systematic surveillance on antimicrobials use and resistance to inform the initial choice of antibiotics upon local antimicrobial resistance profiles. Targeting the other end of the problem, pet owners, our findings indicate the need to educate them on the rational use of antibiotics and, critically, stop antibiotic availability without prescription.