Feed supplements to fish are generally used to overcome any expected diseases and stressors and to sustain eco-friendly fish farming. One of these feed supplements is chitosan, which stimulated growth and immune properties for many aquatic organisms. It is expected that the nano-sized materials may have stronger immune activation in fish than the ordinary size. Therefore, the current study was conducted to evaluate the effect of dietary chitosan nanoparticles (CNP) on growth performance, antioxidant activity, and innate immunity of Nile tilapia, Oreochromis niloticus (L.). Fish (19.8 ± 0.59 g) were fed on diets enriched with 0.0, 0.25, 0.5, 1.0, and 2.0 g CNP/kg diet for 45 days. Fish performance was significantly improved with increasing CNP levels over the control diet with optimum level of 1.0 g CNP/kg diet. Antioxidant-stimulated activity was observed due to dietary CNP supplementation over the control diet in a dose-dependent manner. However, malondialdehyde level decreased significantly, whereas activities of catalase, superoxide dismutase, lysozyme, and respiratory burst increased significantly due to CNP supplementation in a dose-dependent manner. The current study evoked that dietary CNP showed strong immune modulatory properties and enhanced significantly the performance and health of Nile tilapia with optimum level of 1.0 g CNP/kg diet. •Nile tilapia was fed on diets enriched with different CNP levels for 45 days.•Fish performance improved significantly with increasing CNP levels with optimum level of 1.0 g/kg diet.•Antioxidant-stimulated activities were observed with dietary CNP over the control diet especially at high levels.•Dietary CNP enhanced the immune response of Nile tilapia in a dose dependent manner.