Antimicrobial resistance (AMR) has become a major public health threat. Due to the increased risk of bacterial resistance, antimicrobial drugs will lose some effectiveness for all users with each usage. Our aim is to assess students’ and faculty members’ awareness of antimicrobial use and its role in the development of antimicrobial resistance at Taibah University through this cross-sectional study. Between March 2025 and May 2025, 100 participants aged 18-44 from Taibah University in Saudi Arabia were surveyed using an online questionnaire. Data were analyzed using Excel and IBM SPSS Statistics version 23.0 (Armonk, NY: IBM Corp.). The sociodemographic characteristics of the participants were analyzed, and their awareness of antibiotic resistance was assessed using various statistical methods. 92% of students selected that unnecessary intake of antibiotic leads to AMR and 76% students chooses that incomplete course of antibiotic leads to AMR but most of them choose to take antibiotic with physician prescription not over the counter. Nearly half of the respondents think antibiotics can be used for viral infections. Healthcare administrators should take an integrated approach, focusing on the main cause of AMR. Despite having good knowledge about antibiotic use respondents involve in practices which leads to the AMR. People’s behavior and actions regarding antibiotics significantly influence the spread of antimicrobial resistance. This study offered valuable insights into the knowledge and practices related to antibiotic use and antimicrobial resistance among the Taibah University community. In general, a good understanding of antibiotic use and antimicrobial resistance was evident among the surveyed sample. Future research on this topic is still necessary.