Excessive use of silver nanoparticles (AgNPs) due to antibacterial properties can raise concerns about their release into environment and potential toxicity in aquatic organisms. Melatonin has several physiological functions especially antioxidant potential against oxidative stress. The current study was conducted to investigate the potential effects of two doses of dietary melatonin on growth performance, plasma biochemistry, and liver enzyme activity in Nile tilapia (Oreochromis niloticus) juveniles. We also investigated the potential ameliorative effect of melatonin in AgNPs-induced biochemical alterations in tilapia fish. The results showed that melatonin-supplemented diets had no significant effect on growth performance of fish (P>0.05). The liver GPx activity increased in fish fed melatonin-supplemented diets (P<0.05), but the SOD activity showed no significant difference in comparison with the control (P>0.05). The administration of melatonin-supplemented diets reduced the activity of liver MDA compared to the control (P<0.05). Feeding fish with high melatonin-supplemented diet (200 mg kg−1 of diet) decreased the plasma glucose, total protein, and AST levels (P<0.05). The liver GPx and SOD activities were higher in high melatonin-treated fish exposed to AgNPs than the control group (P<0.05). Dietary melatonin decreased the liver MDA activity in AgNPs-exposed fish. The plasma glucose, AST, and ALT levels in melatonin-treated fish exposed to AgNPs decreased compared to the untreated exposed fish (P<0.05). Melatonin-treated fish exposed to 0.05 and 0.5 mg L−1 of AgNPs had lower plasma LDH level than the control group (P<0.05). The results showed that consumption of melatonin-supplemented diets could modulate some of the biochemical indices of plasma and liver in Nile tilapia. The findings also indicated the ameliorative effect of dietary melatonin on AgNPs-induced toxicity in Nile tilapia.