Statement of Problem: Acid producing bacteria including Streptococcus mutans and lactobacilli cause tooth demineralization and lead to tooth decay. Also, oral colonization of the species of Candida has been reported in many studies that are resistant to antifungal agents.Objectives: In this study, antibacterial and antifungal effects of nano-CuO were studied against some oral bacteria and yeast fungi.Materials and Methods: The minimum inhibitory concentrations (MICs) of copper oxide nanoparticles (CuO NPs) for oral bacterial and fungal test strains were determined in 96-well microtiter plate technique. The agar diffusion test (ADT) was employed to assess the antifungal properties of nystatin.Results: The MIC50 value of CuO NPs was determined at the range of 1-10 mg/ml for S. mutans, <1 mg/ml for L. acidophilus, and 10 mg/ml for L. casei. Higher concentrations of CuO NPs (100-1000 mg/ml) were effective on the bacterial cell growth, resulting in 100% reduction in the optical density in TSB medium. The cells of Candida albicans, C. krusei and C. glabrata were treated with CuO NPs and the results showed a decrease in fungal growth at a concentration of 1-1000 mg/ml in TSB medium. The MIC50 value of CuO NPs was determined 1000 mg/ml for three species of Candida. The diameter of growth inhibition zones of 1100 mg/ml nystatin was obtained 15-21 mm for clinical isolates of three species of Candida.Conclusions: With respect to the potential bactericidal activity of CuO NPs on various cariogenic bacteria examined in this study, these NPs could be introduce as a candidate control agent for preventing dental caries or dental infections. In our study, on the other hand, Nano copper oxide had a weak effect on the candida species.