Certain types of nanoparticles, especially zinc oxide nanoparticles (ZnONPs), are widely reported to be capable of the inhibition of harmful bacteria, yeasts, and filamentous fungi. The unique physicochemical and biological properties of ZnONPs also make them attractive to the food industry for use as a promising antifungal agent. This Review thoroughly introduces the preparation methods and antifungal properties of ZnONPs and analyzes their possible antifungal mechanisms. The applicability of ZnONPs in food packaging and nutritional supplements and as an antimicrobial additive is also documented. Moreover, evaluations for biological safety of ZnONPs are objectively reviewed in this paper. The discussions addressed in this Review not only have theoretical significance but also are conducive to the development of food safety, nutrition, and human health. The summarized knowledge and future perspectives outlined here are expected to promote and guide new research toward developing and optimizing the application of ZnONPs as a novel class of antifungal agents to help improve food quality as well as food safety in the near future.