This clinical study investigated if daily immersion in denture cleansers reduces microbial counts on removable partial denture's (RPD) biofilm. Twenty-five RPD wearer volunteers were selected and instructed to complement the hygiene of their dentures by immersing them in an enzymatic peroxide-based denture cleanser (Polident® 3 minute) once a day for 3 min for a period of 15 days. The biofilm was collected from RPD surfaces with a swab immediately before (baseline) and after the experimental period. The samples were placed in sterile saline solution, sonicated at 7 W and then plated on specific culture media to quantify total microorganisms, total streptococci and Candida spp. counts. Data from both collections were compared by paired t-test (α=0.05). It was observed a significant reduction on total microorganisms' counts in RPD biofilm after denture cleanser use (p=0.007). This reduction was also observed for total streptococci (p=0.0428), but no difference was observed on Candida spp. counts. 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