Zinc nanoparticles are documented to be harmful to fish because their accumulation in fish bring about many irreversible changes in their health. Nigella sativa and its oil have been endorsed in aquaculture to improve fish health. Two hundred seventy experimental fish (113 ± 5 g body weight) were divided into 6 groups G1–6; control fish fed a diet without any treatment (G1), 0.3% of NSO (G2), 0.5% of NSO (G3), ZnO NPs (40 mg/kg diet) (G4), 0.3% of NSO and ZnO NPs (40 mg/kg diet) (G5), 0.5% of NSO and ZnO NPs (40 mg/kg diet) (G6), the trial lasted for six weeks. Growth performance was enhanced in fish received diets containing NSO, final weight (FW), weight gain (WG), daily weight gain (DWG), and relative growth rate (RGR) were significantly increased with lower food conversion ratios (FCR) compared to the control. The hepatic glutathione peroxidase (GPx), catalase (CAT), and metallothionein (MT) were increased in response to ZnO NPs stress and only 0.5% NSO supplementation could ameliorate such increment. The immune-related genes interleukin1-beta (IL-1β), tumor necrosis factor-beta (TNF-β), transforming growth factor-beta 2 (TGF-β2) and C-type lysozyme as well as growth-related gene insulin-like growth factor 1 (IGF1) in liver showed an upregulation in fish fed with NSO diets. Administration of ZnO NPs lowered the resistance of Oreochromis niloticus against bacterial infection with Aeromonas hydrophila and NSO could enhance the immunity in the highest tested concentration (0.5%) (G6). The obtained results implied that NSO could enhance the oxidative and immune status of O. niloticus which could compensate ZnO NPs stress as well as experimental infection of a virulent strain of A. hydrophila. Our results revealed that NSO might increase fish growth and immunity only at a high dose (0.5%). •Zinc nanoparticles could suppress Nile tilapia immunity.•Nigella sativa oil could ameliorate nanoparticles withdrawals.•Cytokines expressions affected by Zinc nanoparticles and Nigella sativa oil supplementation.