Introduction: Antibiotic resistance is a significant public health concern affecting human and environmental health. Antibiotics in agriculture can contribute to developing antibiotic-resistant bacteria, which can spread to humans through contaminated food or water. Preparing our future healthcare workforce by educating current students on antibiotic resistance threats is crucial, and this can be achieved through well-informed medical and science students. Objectives: This study aims to evaluate the knowledge, attitudes, and practices regarding antibiotic use and antibiotic resistance among medical and science students. These students are expected to have roles, either directly or indirectly, in working with antibiotics in healthcare and environmental contexts in the future. Method: The study was conducted in a cross-sectional manner, and the study tool was developed after conducting an extensive literature search. A pilot study assessed the instrument’s reliability, measuring data for internal consistency using Cronbach Alpha, which yielded a score of 0.827, indicating good reliability. Data in this study was analysed using SPSS version 26 with descriptive and inferential statistics. Results: The study included 312 participants, with 154 (49.2%) from the Faculty of Medicine and 159 (50.8%) from the Faculty of Science. Most participants demonstrated sufficient understanding of antibiotics, faculty being factors that influenced their level of knowledge. Meanwhile, faculty affiliation and attendance in training and workshops were associated with their level of practice. Conclusion: The study revealed that the students had a commendable comprehension of antibiotic resistance, with medical students exhibiting marginally higher levels of knowledge and practice than science students. Based on the findings, future research with an interventional approach is recommended to enhance science students’ knowledge and practice concerning antibiotic use and resistance.