This experiment was conducted to evaluate the effects of dietary supplementation of whey powder (WP), Bacillus subtilis (BAS), and their combination (MIX) on growth performance, intestinal morphology, caecal microflora, hepatic gene expression, blood metabolites, and skeletal muscle antioxidant capacity in Japanese quails.A total of 400 one-day-old Japanese quails were randomly distributed to 20-floor pens (4 dietary treatments, 5 replications per treatment, 20 birds per pen). The birds were fed a basal diet (control, CON) or the basal diet supplemented with 40 g/kg WP, 1 g/kg BAS probiotic or 40 g/kg WP plus 1 g/kg BAS probiotic for five weeks. Feed intake was not affected by the treatments at any stage of the trial. However, the WP, BAS, and MIX feed had better weight gain and feed conversion ratio compared to the CON during the entire production period (day 1-35; p < .05). Feeding the WP, BAS, and MIX diets caused no significant difference in morphometric measures in the duodenum, jejunum, and ileum other than the villus height to crypt depth ratio in the ileum (p < .05). The expression of insulin-like growth factor-1 (IGF-1) and growth hormone genes was highly upregulated in the liver of the birds fed the MIX diet (p < .05).Feeding birds with the diets containing WP, BAS, and MIX increased the population of caecal lactic acid bacteria and reduced serum cholesterol concentration compared to the CON diet (p < .05). Likewise, the tested feed additives increased superoxide dismutase and glutathione peroxidase enzyme activities in the thigh muscle (p < .05).No synergistic effect was found between WP and BAS in studied parameters other than IGF-1 gene expression. Improved growth performance of Japanese quails by feeding the WP, BAS, and the MIX feed could be linked to improved absorptive capacity of the small intestine as well as over-expression of anabolic growth factors.In conclusion, WP with or without BAS could be considered as a beneficial dietary supplement to enhance productive performance, gut functionality, and antioxidant capacity of Japanese quail. K E Y W O R D S antioxidant capacity, enteric ecosystem, Japanese quail, probiotic, whey powder | 887 JAZI et Al. How to cite this article: Jazi V, Farahi M, Khajali F, Abousaad S, Ferket P, Assadi Soumeh E. Effect of dietary supplementation of whey powder and Bacillus subtilis on growth performance, gut and hepatic function, and muscle antioxidant capacity of Japanese quail.