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Abstract Background Antibiotic over-prescribing has generally been considered to be common in Greece, however not much is known about current antibiotic use. Findings The aim of this study was to investigate antibiotic prescribing in a well-defined rural population of 159 adults and 99 children over a 12-month period in Crete, Greece. The daily-defined doses (DDD) for 1000 people/day (DID) were 22.1 and 24.2 for children and adults respectively. The overall DID was 23.4, markedly lower than that previously reported for Greece. The use of penicillins was 49.5% of DDD in children and 31.7% in adults. Quinolones represented 2.2% of the total antibiotics (0% in children). Prescriptions of antibiotics were more common during the 3-month period from January to March for both children and adults. Conclusions The findings of this study confirm the seasonal distribution of antibiotics used and the predominance of prescribing for respiratory tract infections. In the area of the study, antibiotic use seems to be lower than that previously reported for Greece, probably as a result of the recently established net of well-trained primary health physicians.