There is a growing concern nowadays over the exposure of nanomaterials and their effects in aquatic life. In spite of reporting the changes in physiology, reproduction and behaviour in fish by different nanoparticles, the molecular events underlying in the aquatic bodies due to the toxicity of zinc oxide nanoparticles (ZnO NPs) are mainly unexplored. Therefore, the present study carried out an ex vivo exposure of ZnO NPs at various concentrations (0.382, 0.573 and 1.146 mg L-1) in freshwater fish Cyprinus carpio to investigate the potential adverse effects. The results revealed that ZnO NPs exposure altered the haematological parameter and induces the reactive oxygen species (ROS) that leads to elevation of superoxide dismutase (SOD), catalase (CAT), glutathione peroxidise (GPx), glutathione S-transferase (GST) and reduced glutathione (GSH) activity in C. carpio. Furthermore, histopathological analysis exhibited that the ZnO NPs caused lamellar fusion, aneurism, cytoplasmic vacuolation, nuclear alteration, necrotic muscle fiber and pyknotic nuclei in the gills, liver and muscles of C. carpio. ZnO NPs exposure significantly up-regulated the overlapping expressions of SOD1, CAT, GPx1a, GST-α, CYP1A, and Nrf-2 genes. A higher level of Zn bioaccumulation was observed in the following order: gill (35.03 ± 2.50 μg g-1), liver (5.33 ± 0.73 μg g-1) and muscle (2.30 ± 0.20 μg g-1) at 1.146 mg L-1 exposure of ZnO NPs. Hence, the current study indicated that the biogenic ZnO NPs generate toxicity in fishes by modifying the antioxidant defense mechanisms, histomorphology, and oxidative stress encoding genes.