There is evidence that the transmission of Staphylococcus aureus and methicillin-resistant S. aureus between the community and environmental surfaces still exists. Even the means of this transmission remain uncertain, the public transport system may serve as a potential source of different bacteria, and the contact with contaminated public surfaces may increase the risk for bacterial diseases emergence. This study aimed to investigate S. aureus contamination on Bratislava’s public transport vehicles. Forty samples of hand-touched surfaces were collected during December 2015 and March 2017 by using surface sampling method. S. aureus was detected in all analysed swabs. Simultaneously, antibiotic resistance of S. aureus from swabs was evaluated. Of 40 samples, only 23 % did not contain S. aureus resistant to some of 10 analysed antibiotics. On the other hand, the severe prevalence of highly resistant S. aureus to penicillin, methicillin, ampicillin, and cefoxitin was confirmed. 15 % of isolates displayed resistance to at least three antimicrobial classes. The amount of S. aureus was not significantly influenced by the lines or by the analysed surface (grabs rails or on-board stop buttons). However, there was a statistically significant effect of year period, both between samples from December and March and between samples from the same month but different year. The study confirmed the widespread occurrence of resistant S. aureus in public transport vehicles in Bratislava, Slovakia.