Background: Fish health and disease management is the key factor in modern aquaculture where intensification manifolds the infestation chances of parasites. Irrespective of antibiotics, the use of dietary additives gaining attention in development of aqua feed. Vitamin D is an important dietary additive for fish in terms it can boost up the immune system of fish and unluckily fish cannot synthesize vitamin D itself. Methods: The present study was planned to determine the effect of vitamin D as feed additive on the immunity, hematology and body composition of Labeo rohita. Four diets (T1, T2, T3, T4) containing different concentration of vitamin. D (250mg, 500mg, 1.0g and 1.5g) in food and a control diet were administrated for 90 days trial. Result: At the termination of experiment there were significant differences in growth and hematological parameters between control and test diets. T3 (1.0g of vitamin D) showed the maximum weight gain with lowest FCR value (1.43±0.33) however, the RBC count of T4 group was highest (1.85±0.07) than the control, T1, T2 and T3 groups. The body composition of Labeo rohita growth factor and survival rate were significantly higher (P is less than 0.05) in fish fed on diets containing vitamin D at 1.00g/Kg concentration.