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Introduction: Very few studies have been conducted on antibiotic consumption in Madagascar. The objective of this study was to describe antibiotic consumption in a tertiary university hospital in Antananarivo, Madagascar to more clearly define good antibiotic use strategies., Materials and Method: A one-day prevalence survey was conducted on 5 April 2011 in the 339-bed Befelatanana Hospital with a bed occupation rate of 65.5%., Results: The prevalence of antibiotic therapy among the 222 patients hospitalised on the day of the survey was 57.2%. Almost one half (49.6%) of patients received a combination of at least two antibiotics. The classes of antibiotics most commonly prescribed were beta-lactam antibiotics (55.3%), imidazoles (14.9%), aminoglycosides (14.9%), quinolones (7.9%) and macrolides with 7% of prescriptions. Penicillins accounted for 55.5% of all beta-lactam antibiotic prescriptions, followed by third-generation cephalosporins. Children under the age of 14 years (p<0.019) and patients with invasive devices (p<10(-6)) received more antibiotics. Antibiotic prescription in the Emergency Room–Intensive Care Unit was significantly higher than in the other wards (p<0.01)., Conclusion: A high rate of antibiotic use was observed in Befelatanana University Hospital. Beta-lactam antibiotics were the agents most commonly prescribed and the Emergency Room–Intensive Care Unit was the leading antibiotic prescriber. It is essential to set up a good antibiotic use policy.