Introduction: Antimicrobial resistance is a serious global issue andirrational use of antimicrobial agents against infectious diseases is the main reason for the development of resistance. Containment of antimicrobial resistance requires change in the antimicrobial prescribing behavior of health workers.WHO has recommended training for medical students regarding the prudent prescription of antibiotics. Aim and Objective: To determine the knowledge, attitudes, and practices of medical students and interns regarding the use of antimicrobials. Material and methods: It was a cross-sectional observational study conducted in department of Pharmacology of Shri Guru Ram Rai Institute of Medical and Health Sciences (SGRRIM & HS) Dehradun. Medical undergraduate students from second year to final year and interns were included in the study. Informed consent was obtained from each participant. A pretested, validated google form questionnaire was used in this study. Comparison of quantitative variables between the study groups was done using ANOVA. For comparing categorical data, Chi square (χ2) test was performed and fisher exact test was used when the expected frequency is less than 5. Results: A total of 416 responses were included in the study.The results revealed that interns have better knowledge compared to other MBBS students. The ANOVA test indicated a statistically significant difference in the knowledge score between student groups (F = 9.05, p = 0.001).Only59.9% of students and interns had correctly answered the question related to use of antibiotics in treatment of diarrhea and viral fever.Additionally, 87% of participants agreed with the statement emphasizing the importance of large-scale 'antimicrobial awareness programs' for spreading information about these medications.It is worth noting that a majority (80%, n=333) always check for expiry dates before using any antimicrobial product. Conclusion: The study sheds light on the existing gaps in the knowledge, attitude, and practice of antimicrobial use among interns and medical students.Targeted interventions and educational initiatives that aim to foster responsible antimicrobial prescribing behavior and mitigate the growing threat of antimicrobial resistance must be the part of medical education. Antimicrobial knowledge should beimparted as integrated approach and should be considered as a certifiable skill for the successful completion of internship.