Background: Irrational use of antimicrobials and inaccurate practicing behavior leads to the issue of antibiotic resistance. This can be tackled by spreading awareness with the assistance of future medical practitioners. Hence, the present study was taken to determine the knowledge, attitude, and practicing behavior regarding antimicrobial use and awareness of antimicrobial resistance among interns and postgraduates.Methods: The study was conducted on interns and postgraduates in a tertiary care hospital. A standardized questionnaire was distributed to 120 participants and Ethical approval was taken before the study. Data was analyzed using SPSS software. For data comparisons, Chi-square tests were used, p≤0.05 is considered significant.Results: The study showed a majority in the 25 to 34 years of age group with female predominance. The subject of antibiotic resistance was moderately relevant to a majority, which highlighted a need for an increase in awareness workshops. The actual knowledge of respondents seems to be accurate in the study, a fair amount of attitude and practicing behavior was observed as well. A significant difference between the interns and postgraduates in knowledge (χ2=13.736, p=0.03), attitude (χ2=68.091, p=0.01), and practice (χ2=34.821, p=0.01) were noticed.Conclusions: Accurate knowledge and practicing behavior were observed in postgraduates and a fair attitude was observed in interns towards antimicrobial use and resistance. The awareness must be extended regarding this issue by conducting educational programs and by updating guidelines. Advancement in antibiotic prescribing pattern is required.