BACKGROUND: Antibiotic resistance is an escalating global health issue, with particularly severe implications in low- and middle-income countries (LMICs) such as Nigeria. This study examines antibiotic-resistant bacteria's prevalence and molecular characteristics in daycare centres in Ile-Ife, Nigeria, where high antibiotic use and limited infection control measures present significant challenges. METHODS: Between November 2017 and July 2019, samples were collected from 20 daycare centres, including swabs from fomites and children. Bacterial isolates were identified and assessed for antibiotic susceptibility using standard methods. Molecular techniques, including PCR, were employed to detect resistance genes such as RESULTS: The study found high resistance levels among common pathogens, with CONCLUSIONS: The results highlight the critical need for improved surveillance, targeted antimicrobial stewardship and enhanced infection control practices in daycare centres to address the growing threat of antibiotic resistance. This research offers valuable insights into resistance dynamics in paediatric settings and supports the development of strategies to manage the spread of resistant bacteria in LMIC contexts.