Natural substances are now generally preferred over chemical and synthetic compounds for the growth and immune enhancement of aquatic organisms. The aim of this study was to evaluate the effect of Saccharomyces cerevisiae extract and hydrolyzed powder on immunity, hematological parameters and body composition in rainbow trout, Oncorhynchus mykiss. Six hundred rainbow trout (50 ± 5 g mean weight) were acclimated to laboratory conditions and then randomly divided into four groups of triplicate tanks. The first group was fed with a commercial diet (control) without supplementation. The second and third groups were given a diet supplemented with 1% of yeast extract and hydrolyzed powder, respectively. The fourth group was also fed with a basal diet supplemented with 0.5% of both substances. Fish were cultured in 300-L polyethylene tanks for 60 days; immune and hematological parameters, fillet composition and disease resistance were analyzed at days 0, 30 and 60. Results showed that a combination of Saccharomyces cerevisiae extract and hydrolyzed powder could improve the immunity and alter hematological parameters of the rainbow trout compared to the control. Mortality rates of fish fed yeast extract and hydrolyzed powder were also lower than in fish fed the control diet after challenging with Yersinia ruckeri. There were no significant changes in rainbow trout fillet composition compared to the control. It can be concluded that fish diet supplementation with a mixture of yeast extract hydrolyzed powder is preferable compared to each one used alone.