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Background and Objectives: Zinc oxide (ZnO) nano-particles have been proven to have strong antibacterial activity against food borne pathogens. The practical applications of different concentrations (Control, 0, 1, 2, 4, 6 and 8 mM) of ZnO suspensions containing 1% acetic acid were investigated against the pathogenic bacteria like Staphylococcus aureus. Materials and Methods: The projects of laboratory and field number 3 were repeated. The order of 1% acetic acid, and the 6 mM and 8 mM suspension concentrations of zinc oxide nanoparticles containing acetic acid was/were used as antibacterial juices. The experiments were repeated three rounds. The data were analyzed by ANOVA software, and the P<0.05 level of significance was identified. Results: The results also exhibited that ZnO and acetic acid had inhibitory effect on the growth of all strains during the 24 h culture period in mango juice, as compared to the control experiment, which was further confirmed in the liquid culture. Conclusions: This is the first report describing the antibacterial activity of ZnO NPs in mango juices that showed the potential of these nano-particles for use as an antibacterial agent in the food industry. Keywords: Antibacterial, Acetic acid, Food, Nano-particles, Mango juice, Zinc oxide