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The aims of this present study were (1) to assess the antimicrobial effect of ozone from a novel ozone–generating device (Heolozone, USA) [0.052% (v/v) in air delivered at a rate of 13.33 ml·s^–1] on primary root carious lesions (PRCLs) and (2) to evaluate the efficacy of ozone specifically on Streptococcus mutans and Streptococcus sobrinus. In study 1, 40 soft PRCLs from freshly extracted teeth were randomly divided into two groups to test the antimicrobial effect on PRCLs from exposure to ozonated water for either 10 or 20 s. Half of a lesion was removed using a sterile excavator. Subsequently, the remaining lesion was exposed to the ozonised water for a period of either 10 or 20 s (corresponding to 0.069 or 0.138 ml of ozone, respectively). Using paired Student t tests, a significant (p<0.001) reduction (mean ± SE) was observed in the ozone–treated groups with either a 10–second (log₁₀ 3.57±0.37) or 20 …