In the present study, the antimicrobial properties of a new ozonized olive oil (O-zone gel) against oral and periodontal pathogens will be evaluated and compared with that of common CHX-based agents. O-zone gel was compared with two agents based on chlorhexidine digluconate (CHX): Corsodyl Dental Gel and Plak Gel. A. actinomycetemcomitans, P. intermedia and S. mutans, were selected and the antibacterial capability of the compounds was tested by using direct contact agar diffusion test (DCT) and minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) evaluations. Differences between specific means were analyzed by a one-way analysis of variance (ANOVA). Group means were compared using a one-way ANOVA and Tukey?s test (P<0.05). O-zone gel reported inhibition zones which correspond to 33% and 43% of that achieved by the CHX agents. No inhibition of bacterial growth (MIC) on the Gram-positive strain by using O-zone gel was found and no antimicrobial effect (MBC) was observed by using O-zone gel on both Gram-negative and -positive strains. The new ozonated oil was a relatively moderate antiseptic. Gram-negative bacteria proved to be more sensitive to ozonized olive oil than Gram-positive ones. The ozonized olive oil demonstrated a lower antibacterial activity if compared to the CHX-based agents tested.