Tilapia (Oreochromis niloticus) was exposed to different sizes of zinc oxide nanoparticles (ZnO-NPs) to evaluate their organ pathologies (kidney, liver, gill, and intestine), osmoregulatory responses and immunological parameters. Sub-chronic exposure was conducted in fresh water with 1 and 10 mg/L concentrations of the small (10-30 nm) and large-size ZnO (100 nm) particles for 7 and 14 days. In this study, it is found that small and large forms of ZnO-NPs cause various pathologic findings in the target organs at all concentrations. These findings are increased of melanomacrophage aggregates, tubular deformations, necrosis and cytoplasmic vacuolations in the kidney, oedema, mononuclear cell infiltrations, fatty changes, pyknotic nuclei and hepatocellular vacuolations in the liver, hyperplasia, aneurysms, and epithelial liftings in the gills, and hyperplasia, swelled of goblet cells, villus deformations in the intestine. Results showed that respiratory burst and potential killing activity at the small-size ZnO concentration significantly increased compared to the control group (p < 0.05) but significant reductions of these parameters at the large-size ZnO concentrations compared to control (p < 0.05) were measured. These findings demonstrate the potential of each particle size to cause significant damage to the immune system. Moreover, because ZnO NPs inhibit the Na, K-F-ATPase activity at all concentrations and increase serum Ca2+ and Cl- levels especially in gill, these particles are osmoregulatory and toxicant for tilapia fish. As a summary, both sizes of the particles have led to organ damage, osmoregulatory changes and immune disorder in tilapia fish. (C) 2015 Elsevier Ltd. All rights reserved.