In this work, the effects of two different amounts of zinc oxide (ZnO) nanoparticles on the antimicrobial activities of poly (vinyl chloride)-based films to inactivate food pathogens were investigated. Results showed that the ZnO-coated film exhibited a good inhibition effect on the growth of Escherichia coli and Staphylococcus aureus and its ability was attributed to the ZnO nanoparticles. The ZnO-coated films exhibited more effective antibacterial activity for S. aureus. However, antifungal activity of the ZnO-coated films (20 mm × 25 mm) against Aspergillus flavus and Penicillium citrinum was not observed. It is likely due to the complexity of the fungal cell wall and the ZnO nanoparticles without UV light irradiated or the insufficient amount of nanoparticles. Our findings reveal that ZnO nanoparticles have a good potential to be coated on a plastic film to make antimicrobial packaging against bacteria such as E. coli and S. aureus.