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Introduction: Public knowledge and attitudes towards antibiotics play a vital role in the success of the treatment process. This study aimed to assess public knowledge and attitudes toward antibiotic usage which could serve as baseline data for future studies within a government hospital setting in Malaysia. Methodology: A self-administered cross-sectional survey involving 408 respondents was conducted using a validated questionnaire at an outpatient pharmacy department in Penang Hospital, Malaysia, from February to March 2009. Results: Nearly 55% of the respondents had a moderate level of knowledge. Three quarters of the respondents (76.7%) could correctly identify that antibiotics are indicated for the treatment of bacterial infections. However, 67.2% incorrectly thought that antibiotics are also used to treat viral infections. About 59.1% of the respondents were aware of antibiotic resistance phenomena in relation to overuse of antibiotics. With regard to attitudes, 38% believed that taking antibiotics when having cold symptoms could help them to recover faster, while 47.3% expected antibiotics to be prescribed for common cold symptoms. Age, race and educational level were among the demographic characteristics significantly associated with knowledge and attitudes toward antibiotic use. Poor level of knowledge was found in less than one-third of the respondents whereas more than one-third of the respondents wrongly self-medicate themselves with antibiotics once they have a cold. Conclusion: Educational interventions are needed to promote prudent use of antibiotics among the public. [ABSTRACT FROM AUTHOR]