The present study evaluated the effects of dietary inulin on growth performance, body composition, serum, biochemical, and mucus immune factors; as well as innate immune responses of rainbow trout fry challenged with Aeromonas hydrophila. Four diets were prepared using a commercially available fish feed as a basal diet and different levels of prebiotic inulin incorporation; 0 (control), 1, 2, and 3%; referred to as C, T1, T2, and T3, respectively. The findings of the 60-day feeding trial showed that inulin inclusion affected final weight, food conversion rate (FCR), and specific growth rate (SGR) compared to that of the control group (P < 0.05), in which the lowest FCR was observed in T3. Body composition analysis revealed that inulin significantly increased protein content and decreased lipid levels, especially in the T1 and T2 groups. The lowest ash level was noticed in T2 (P < 0.05). Blood total protein, albumin, globulin, cholesterol, and glucose were not affected by inulin supplementation (P > 0.05). Analysis of humoral immune responses showed that the inulin supplements significantly increased lysozyme and complement activities (P < 0.05), as well as higher red blood cell count (RBC) and hemoglobin (Hb) in fish, fed 2% inulin, while no significant differences were observed among other treatments (P > 0.05). The mucosal parameters; including lysozyme, alkaline phosphatase (excluding ACH50); protease activities; and total Immunoglobulin (IgM) improved significantly (P < 0.05), particularly in the T2 group. The T2 group also demonstrated the highest survival rate among all groups. The present findings indicate that dietary administration of inulin promotes growth and biochemical parameters, as well as serum immunity and mucosal immune responses of rainbow trout, in which a 2% inclusion produced the best results.