The current work was planned to assess the effects of dietary microalga, Nannochloropsis oculata (NP) on the growth indices, serum biochemistry, non-specific immunity, and resistance of Nile tilapia (Oreochromis niloticus) juveniles against challenge with pathogenic Aeromonas veronii. Fish (10.21 ± 0.28 g) were randomly divided into four treatments in triplicates and were fed on diets supplemented with different levels of NP (0, 5, 10, and 15%) for eight weeks. The results showed that the dietary 5% NP significantly improved the growth parameters (final body weight, weight gain, specific growth rate, and total feed intake) (P < 0.05). Meanwhile, the feed conversion ratio was significantly decreased in NP-supplemented groups with respect to the control group. The serum protein profile (total protein, albumin, globulin, and albumin/globulin ratio) was significantly decreased by 15% NP supplemented group. Meanwhile, liver enzymes (alanine transaminase, and aspartate transaminase) activities were not significantly affected by NP dietary supplementation (P < 0.05). Serum lysozyme activity, nitric oxide (NO), and nitroblue tetrazolium (NBT) levels were significantly enhanced only in 5% NP supplemented group (P < 0.05). Interestingly, the hepatopancreatic and intestinal tissues had apparently normal histomorphology of Nile tilapia fed at 5% NP-supplemented diets. Significant upregulation of cytokines [interleukin 1beta (IL-1β), interleukin 8 (IL-8), interferon-gamma (IFN-γ), transforming growth factor-beta (TGF- β), and tumor necrosis factor-alpha (TNF-α)], with significant downregulation of the antioxidant gene [superoxide dismutase (SOD)] in 10% and 15% NP supplemented groups. Attractively, the relative level of protection (RLP) against challenge with pathogenic A. veronii was significantly elevated in 10% and 15% NP supplemented groups. Conclusively, the obtained results reflect the beneficial roles of dietary NP to improve growth and boost the immune responses of Nile tilapia.