Objectives Nurses are the main communication link for healthcare messages to hospitalized patients and also play a crucial role in preventing the emergence and spread antibiotic resistant bacteria through antibiotic stewardship and infection control programmes. This requires them to possess correct knowledge and attitudes towards antibiotic use and resistance. This study was carried out to identify the level of knowledge, attitude and practices on antibiotic use and antibiotic resistance among student nurses. Results A descriptive cross-sectional study was conducted using a pre-validated, self-administered questionnaire with closed and open ended questions, among 199 student nurses at a government nurses training school in Sri Lanka. Scores and proportions were analysed with non-parametric methods and thematic analysis was done for the qualitative data. The study cohort had a mean knowledge score of 71.9% (SD 14). However, close to 40% believed that taking antibiotics will help to prevent cold from worsening and make recovery faster. Infection control was identified as the main method that nurses can engage in preventing antibiotic resistance. While the knowledge among our study cohort appeared to be good, some misbeliefs were present. Our findings can be used in developing the nursing curricula on antibiotic use and resistance.