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Adem Kose,1 Cemil Colak2 1Department of Infectious Diseases and Clinical Microbiology, Faculty of Medicine, Inonu University, Malatya, Turkey; 2Department of Biostatistics, and Medical Informatics, Faculty of Medicine, Inonu University, Malatya, TurkeyCorrespondence: Adem KoseDepartment of Infectious Disease and Clinical Microbiology, Faculty of Medicine, Inonu University, Elazig Yolu 10. Km, Battalgazi, Malatya, 44280, TurkeyTel +90 4223410660/ 4405Fax +90 4223411220Email adem.kose@inonu.edu.trBackground: We aimed to evaluate the level of theoretical knowledge of rational antibiotic use and awareness about emergence of multidrug resistance (MDR) among the senior students at a medical school and the actively-working family physicians.Methods: This cross-sectional research was carried out between 1 February and 30 April 2019 in Malatya province. Two-hundred twenty-five senior students in the Inonu University Medical School (Group 1) and 230 actively-working family physicians in Malatya primary health care services who were found to be eligible (Group 2) were included in this study. Power analysiscalculated the minimum of 240 participants with a proportion difference of 0.18 between the groups, a type I error of 0.05 and a type II error of 0.20. A questionnaire including seven sections and 28 questions was applied to the participants.Results: Doctors were more hesitant in pre-graduation and more self-confident in the post-graduation period for the decision to start antibiotic treatment. In addition, doctors forget their theoretical knowledge of antibiotics over time and are unable to follow current developments after graduation. The most important concern in the pre-graduation period was the choice of antibiotics from the wrong group, while in the post-graduation period it is the fear of the presence of unproven infection.Conclusion: Physicians’ antibiotic prescribing habits, attitudes and behaviors vary before and after graduation. Sustainable education for antibiotic use for physicians after graduation can contribute positively to reduce of antimicrobial resistance rates and to increase awareness about the use of rational antibiotics.Keywords: antimicrobial resistance, multidrug resistance, knowledge, awareness, rational antibiotic use, MDR