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Background: Antibiotic resistance is an important public health issue. This study sought to measure general practitioners’ (GPs) knowledge, attitude, and practice on antibiotic resistance; explore the adjusted associations between their antibiotics prescribing practice score and knowledge and attitude scores, and identify barriers for rational prescription of antibiotics by GPs. Methods: A cross-sectional self-administered survey was conducted among GPs. Multivariable linear regression was applied to analyze the data. Results: Overall, 291 GPs participated in the study. Knowledge, attitude, and practice mean percent scores were 58.3%, 67.5%, and 63.0%, respectively. In the adjusted analysis, the practice percent score was significantly associated with the attitude percent score (= 0.58; 95% CI: 0.41, 0.75; p < 0.001), though the relation between the practice and knowledge scores was insignificant. The main barriers reported by GPs: lack of rapid diagnostic tests, high costs of laboratory tests, high costs of some antibiotics, and lack of guidelines. Conclusions: Identified low knowledge, attitude, and practice mean percent scores suggest a need for improvements in these areas. Availability of rapid and inexpensive diagnostic tests, enforcement of prescriptions could potentially prevent the development of antibiotic resistance. Key messages: Availability of rapid and inexpensive diagnostic tests, enforcement of prescriptions, and regulation of antibiotics’ prices could help to improve antibiotics prescribing practices of GPs. Low knowledge, attitude and practice levels regarding antibiotic resistance among GPs in polyclinics suggests a need for improvement in these areas. [ABSTRACT FROM AUTHOR]