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To investigate knowledge and perceptions about antibiotic prescription in two university hospitals.Physicians completed four case vignettes describing infections and a questionnaire. For each vignette, the physicians were asked to determine whether hospital admission and antibiotic treatment were needed; whether a treatment change was needed; and the duration of antibiotic treatment. The questionnaire collected data on beliefs and perceptions regarding antibiotic prescription.Of 412 eligible physicians, 206 agreed to participate. Factors associated with a vignette score above the median were anaesthesiologist/intensivist (adjusted odds ratio, 3.09; P = 0.02), perception of inappropriate antibiotic use as risky for the patient (adjusted odds ratio, 2.84; P = 0.03) and self-efficacy (adjusted odds ratio, 2.18; P = 0.02), whereas being a surgeon was associated with a vignette score lower than the median (adjusted odds ratio, 0.14; P < 0.0001).The high participation rate suggested awareness of antibiotic use. Educational programmes specifically targeted at surgeons are needed. We identified cognitive factors that affect knowledge of antibiotic prescription, and may help in the design of education programmes and interventions aimed at improving antibiotic use.