The goal of this study was to determine the role of public transport in the transmission of microorganisms. A total of 60 swab samples were collected in the morning and evening from handles in public transport trams, metrobuses, and buses. Swab samples were examined using microbiological methods, and the number and types of microorganisms were determined. Total aerobic bacterial and fungal counts in samples collected in the evening were higher than those in samples collected in the morning from trams and metrobuses. However, the total bacterial and fungal counts were very high in samples collected in the morning and evening from buses. Staphylococcus aureus, coagulase-negative staphylococcus, and Enterococcus spp. were isolated from these samples. The results of our study show that public transportation can be a significant reservoir for spreading pathogenic microorganisms. For this reason, it is very important to regularly follow cleaning and hygiene rules and to inspect these vehicles.