Three hundred Oreochromis niloticus were divided into five groups; the control fish (CNT) were fed a basal diet, beta glucan (βG) group was fed 0.5 g/kg−1 βG, atrazine (ATZ) group was exposed to 1/5 96‐hr LC50 (1.39 mg/L) ATZ, the (βG/ATZ) group was fed βG while being exposed to ATZ, and the (βG then ATZ) group was supplemented with βG for fifteen days before exposed to ATZ. ATZ exposure caused a decline in growth that was ameliorated by βG. ATZ reduced the levels of total and different types of leucocytes. Additionally, ATZ exposure caused reductions in total proteins, globulins, α 1‐globulin, α 2‐globulin, ɤ‐globulin, immunoglobulin M, lysozymes, superoxide dismutase, nitric oxide and catalase but increases in hepatic transaminases and malondialdehyde without any variations in albumin and β‐globulin. Exposure to ATZ also resulted in a rise in the mRNA level of IL‐8. In contrast, expression of IgM, SOD and CAT were decreased in the tilapias exposed to ATZ. Exposure to ATZ increases the susceptibility response to Aeromonas sobria challenge, as indicated by an increase in cumulative mortality post‐challenge. Supplementation with βG fifteen days before (βG then ATZ group), counteracted the adverse effects of ATZ on the immune, biochemical and antioxidants values, though only slight alleviation was observed with simultaneous treatment (βG/ATZ group). Our results established that ATZ has adverse impacts on immune responses, antioxidant equilibrium and its related genes. While, supplementation with βG before exposure to ATZ may be valuable for counteracting the possible damage caused by ATZ water pollution than its simultaneous treatment with ATZ.