A feeding trial was conducted for 90 days in glass aquaria to check the effect of exogenous enzyme (α-amylase) added feed on growth performance, health and hematological analysis of Pangasianodon hypophthalmus (initialaverage weight 32.46±0.36 g). The 40% crude protein diet was prepared by adding feed ingredients along with 0.75, 0.50, 0.25, 0.00 g kg-1 of α-amylase as T1, T2, T3 and T4 (control). At the end of feeding trial, 100% survival rate was recorded. Growth rate was significantly increased in fish fed with enzyme supplemented diets in comparison with control group. The maximum increase in growth rate was recorded in treatment # 2. Highest protein contents 68.18% was observed in treatment # 3 (0.75g kg-1 α-amylase). Specific growth rate (SGR), feed conversion ratio (FCR) and condition factor (CF) were higher in all enzyme supplemented groups as compared to control group (p<0.05). Hematological parameters were recorded among all groups and concentration of red blood cells (RBCs), white blood cells (WBCs), hematocrit (HCT) and numbers of lymphocytes were highest in treatment # 1 with the mean values of 1.29×10ˆ12 L-1, 54.9×10ˆ9 L-1, 18% and 48.7×10ˆ9 L-1. Microbial load was also determined inside and outside of fish as highest microbial load was observed in control group fish on both media nutrient agar (NA) and eosin methylene blue (EMB) viz. 1.8×103 and 1.56×103 (cfu ml-1) in gut contents whereas on skin only total bacterial count (0.36×103)was observed and no coliform bacteria were present while least CFU’s were counted in group 2 among skin 0.14×103 cfu ml-1 and gut 0.10×103 cfu ml-1 on nutrient agar. The results suggested that the enzyme supplementation improved the growth and health of P. hypophthalmus.