Background: It is described in the literature that between 25% to 50% of antimicrobial prescriptions, in the hospital environment, are unnecessary or inadequate, directly impacting antimicrobial resistance. The National Health Surveillance Agency launched the “Brazil Stewardship Project” to assess the national panorama of antimicrobial use management programs, therefore, it is expected that managers will be encouraged to implement the antimicrobial use management program, with the publication of the results, enabling the comparison of national data. In this context, measures and indicators of antimicrobial use will be particularly important. We demonstrate the application of the indicators DOT, LOT e DOT/LOT ratio to explore and monitor antimicrobials use. Methods: This is an observational, analytical and prospective study conducted in a teaching hospital, with 94 active beds, distributed between the intensive care unit (ICU), the surgical clinic (SUR), the medical clinic (MED), the pneumology/infectology (PNE/INF) and pediatrics (PED). Results: During the study period, a total of 11,634 patient-days were followed up and an average of 50.4% of patients were exposed to some antimicrobial, showing significant reduction throughout the year. In 376 days, for every 1,000 days of hospitalization, patients were receiving antimicrobial therapy (LOT = 376/1000pd). Overall, the 1st generation cephalosporins and fluoroquinolones were highlighted both in the number of prescriptions and in the duration of therapy. The calculated global DOT / LOT ratio showed that each patient received an average of 1.5 antimicrobials during the hospital stay. The incidence of antimicrobial resistance, globally, for both Methicillin R and Carbapenem R , was 1 per 1000 patient-days. Conclusions: The indicators addressed in this study proved to be of great relevance for obtaining data regarding the use of antimicrobials, contributing to the initiatives to be taken by the Stewardship program. This study is expected to encourage other hospitals to monitor the use of antimicrobials, allowing comparison of data.