Bacteriological air sampling was conducted at 25 dental units during restorative treatment sessions before and after disinfection of dental unit waterlines (DUWL) with hydrogen peroxide. Air samples for determining the concentration and species composition of aerobic and facultatively anaerobic bacteria were collected with the portable Reuter Centrifugal Sampler (RCS Plus) in the dental operation area close to patient’s mouth. Large concentrations of airborne bacteria in the range of 0.35–40.08 × 10³ cfu/m³ (median = 1.63 × 10³cfu/m³) were recorded before DUWL disinfection. After disinfection, the concentrations were significantly lower (p<0.05), ranging from 0.51–3.82 × 10³ cfu/m³ (median = 0.9 × 10 3 cfu/m³). Streptococci were most numerous among airborne bacteria before DUWL disinfection, forming 79.23% of total isolates. The remaining isolates were staphylococci/micrococci (15.7%), corynebacteria (2.3%), endospore-forming bacilli (1.45%), Gram-negative bacteria (1.31%), and actinomycetes (0.01%). After DUWL disinfection, a significant decrease in the numbers of streptococci (p<0.05) and Gram-negative bacteria (p<0.01) was noted, while the numbers of other types of bacteria were unaffected. Altogether, 50 species or genera of bacteria were identified in the examined air samples before and after DUWL disinfection. Of these, 36 species or genera are considered potentially pathogenic, as a potential cause of infection, allergic disease or intoxication. In conclusion, the high pollution of dental operation area with bacteria indicates a need for use of preventive measures protecting dental staff and patients, such as DUWL disinfection that proved efficient in decrease of exposure in the present study.