Dietary zinc oxide nanoparticle (ZnO NP) possessing multiple biol. activities might be a potential substitute for the combination of ZnSO4 and antibiotic growth promoter in broiler diets.Therefore, this study was conducted to investigate the efficacy of dietary ZnO NP as a substitute to the combination of ZnSO4 and xanthomycin for growth, antioxidant capacity, immunity, intestinal barrier and liver function in broilers.Arbor Acres chicks (n = 320) were assigned to 5 treatments with 8 replicates.Birds received a basal diet supplemented with 80 mg/kg ZnSO4 plus 5 mg/kg xanthomycin (ZnSO4 + Xanthomycin) or 0 (neg. control, NC), 40, 80, and 160 mg/kg ZnO NP for 42 days.The average daily gain, average daily feed intake and feed-to-gain ratio showed dose-dependent increases with the increasing level of dietary ZnO NP during the 21-42 day and 1-42 day stages.The final body weight (42 d) and serum concentrations of insulin-like growth factor 1 and growth hormone increased linearly with the increasing level of dietary ZnO NP.In addition, 80 mg/kg ZnO NP increased the serum concentrations of IgA, IgG, IgM, andinterleukin-10 and peroxidase activity, the jejunal mucosal villus height, villus width and goblet cell numbersDietary 80 or 160 mg/kg ZnO NP significantly altered mRNA abundances of genes related to antioxidant status, intestinal barrier and immunity in the jejunal mucosa.These results indicated that dietary supplementation with 40-160 mg/kg ZnO NP caused no obvious neg. effects on liver function, but effectively improved growth performance, intestinal barrier function, immunity and antioxidant capacity.HighlightsDietary 40-160 mg/kg zinc oxide nanoparticle (ZnO NP) improved growth performance and enhanced immunity of broilers without obvious neg. effects on liver function.Dietary 40-160 mg/kg ZnO NP improved intestinal barrier and intestinal morphol., and enhanced antioxidant capacity via Nrf2/HO-1 pathway.Doses of 40-80 mg/kg ZnO NP were suggested to alternate the combination of 80 mg/kg ZnSO4 and 5 mg/kg xanthomycin in diets of broilers.