The present investigation aimed to evaluate the effect of dietary chitosan supplementation on growth performance, body composition, immune response and histopathology of Nile tilapia, and also the in vitro antibacterial activity of chitosan against Streptococcus agalactiae (S. agalactiae). About 180 fish (average body weight 39.3 ± 0.3 g) were randomly divided into three groups according to chitosan supplementation: control group (basal diet without chitosan), Ch3 group (3 g chitosan/ kg diet) and Ch5 group (5 g chitosan/kg diet). Growth performance parameters and body proximate composition were measured before infection but biochemical parameters and lysozyme and antibacterial activities before and after experimental infection. Results of the present investigation showed dietary chitosan (5 g chitosan/ kg diet) significantly (p < .05) improved growth performance parameters, body composition (dry matter, crude protein, ether extract, ash, and carbohydrate) and serum biochemistry (total protein, albumin, globulin, with no effect on AST, ALT, urea and creatinine) before infection in Ch5 group than the control. After infection, liver enzymes (serum AST and ALT) were maintained lower in fish fed Ch3 or Ch5 than the control. Serum lysozyme and bactericidal activities significantly increased (p < .05) in chitosan groups before and after the challenge. The mortality rate was markedly reduced in the Ch3 group and prohibited in the Ch5 group after the experimental infection. In conclusion, feeding 3 or 5 g chitosan/kg diet increased the growth rate and improved FCR of Nile tilapia. In addition, it reduced mortality by its antibacterial and immunostimulant effects. K E Y W O R D S bactericidal activity, growth performance, histopathology, lysozyme activity, Nile tilapia, Streptococcus agalactiae | 1121 FADL et AL.