Staphylococcus aureus is an important zoonotic pathogen of public health concern, frequently associated with foodborne intoxication and infection in poultry systems. This study investigated Staphylococcus load, seasonal distribution, antibiotic resistance, and resistance genes in isolates recovered from three broiler farms in Egypt. A total of 324 samples were collected from cloacal swabs, air, water, litter, feed, handlers, equipment and walls. Litter reported the highest mean S. aureus count (6.3 ± 0.31 log CFU), whereas air samples showed the lowest (3.57 ± 0.40 log CFU). During summer and winter, litter remained the most heavily contaminated source (6.43 ± 0.18 in vs. 6.21 ± 0.37 log CFU, respectively). Overall, 60.8% (197/324) of the examined samples were positive for S. aureus. Cloacal and hand swabs were significantly associated with contamination risk with prevalence rates of 96.26%, (OR = 13) and 94.4%, (OR = 8), respectively. Although, S. aureus prevalence was higher in summer (64.8%) than in winter (56.7%), the difference was not significant (p = 0.139). All S. aureus isolates were resistant to neomycin and clindamycin, while 85.7% were sensitive to meropenem and vancomycin. Most isolates (85.71%) exhibited multidrug resistance (MDR), with a mean MAR index of 0.496; remarkably, the highest value was recorded in air samples (0.844). The resistance genes, ermA and aac(6′)-aph(2″) were detected in 50% and 25% of the isolates, respectively, while mecA and vanA gene were each detected in one of the eight tested isolates(12.5%). In conclusion, the persistence of multidrug-resistant S. aureus in poultry environments appears to be closely associated with inadequate hygiene. These findings emphasize the urgent need for improved sanitation and strict biosecurity measures to mitigate risks to poultry production and public health.