Ozone (O3) is a reactive oxidant increasingly applied in biomedical settings, yet its dose-dependent effects on innate immune cells, particularly those from non-human species, remain insufficiently defined. Within a One Health framework, this study examined how two clinically relevant O3 exposure regimens (30 µg/mL and 90 µg/mL) affect porcine neutrophils and monocytes isolated from peripheral blood. Cell viability, reactive oxygen and nitrogen species (RONS) production, and the activity of key enzymes (myeloperoxidase, elastase, alkaline phosphatase, arginase) were assessed at 1 h and 24 h post-exposure. The lower dose induced mild functional activation without compromising viability, whereas the higher dose triggered pronounced oxidative stress, enhanced degranulation, and reduced neutrophil viability by more than 60%. Neutrophils exhibited a stronger and more dynamic response than monocytes, which retained viability and differentiation capacity at 30 µg/mL but showed impaired function at 90 µg/mL. These findings highlight the dual nature of O3, where controlled exposure may support immunomodulation, while excessive dosing disrupts cell function. Defining safe and effective therapeutic windows remains critical for future applications.