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Background: Staphylococcus aureus is a ubiquitous organism that normally colonizes the human body and causes food poisoning. This study aimed to determine the rate of nasal carriage of S. aureus among food handlers in Sana'a restaurants in, Yemen and performed an antibacterial susceptibility test. Methods: This cross-sectional study was carried out among healthy food handlers from November 2022 to March 2023. Approximately 024 nasal swabs were collected and S. aureus was isolated and identified according to standard bacteriological procedures. Moreover, the antibacterial susceptibility pattern was determined using the disc diffusion method. Additionally, the required data were obtained by using the pretested questionnaire. Results: Of 420 nasal swabs, 78 (18.6%) food handlers were S. aureus nasal carriers. The prevalence rate of S. aureus was significantly higher among subjects aged >30 years (23.1%), males (18.8%), and uneducated subjects (25%). Additionally, a higher rate of S. aureus nasal carriers was observed among food handlers who worked as cooks (19.4%), had less than 2 years of experience (20.5%), and had low hygiene (29%). The S. aureus isolates were sensitive to ciprofloxacin (71.8%), vancomycin (76.9%), and amoxicillin-clavulanic acid (61%). Conversely, the isolated S. aureus was resistant to oxacillin (69.2%) and methicillin (66.7%). Conclusion: These data stress that food handlers with a high S. aureus nasal carrier rate may pose significant risks to consumers. Therefore, creating restaurant policies and rules is necessary to provide safe and healthy food to consumers. [ABSTRACT FROM AUTHOR]