Probiotics are useful bacteria in promoting the development, health status and survival of aquatic animals. This current experiment was focused to check the hemato-biochemical parameters of Nile Tilapia (Oreochromis niloticus) by using probiotic mixture. Fish fingerlings were collected and then acclimatized for 2 weeks in 70 L glass aquaria. The fish feed was prepared by mixing different probiotics which include Bacillus subtilis, B. megaterium, B. pumilus, B. licheniformis and Enterococcus faecalis in the basal diet and the trial was carried out for 90 days. Four treatments were designed for trial, Control T0 (without probiotic), T1 (2 g probiotic), T2 (4 g) and T3 (6 g). Each aquarium contained equal fish with three replicates and fed according to 5% of their body weight twice a day. Physico-chemical parameters of water were monitored. After each month in the trial, the hemato-biochemical parameters were measured. The value of RBCs, Hb, Hct and PLT increased in T2 with 4 g probiotic instead of other treatment groups. WBCs were increased in T3 group. While in MCV, MCH and MCHC a significant decrease was observed. In biochemical parameters which include Total Protein, Albumin and Globulin, a significant increase in T3 was observed. The current investigation showed that probiotics improve the hematological and biochemical status of Nile Tilapia.