ABSTRACT Effects of dietary supplementation of Bifidobacterium longhum, Bifidobacterium thermophilum, Bacillus subtilis, and Lactobacillus acidophilus on growth performance, health condition, body chemical composition, and water quality of Nile tilapia (Oreochromis niloticus) fingerlings were evaluated. Five experimental treatments in triplicate were fed a diet containing 30% crude protein supplemented with variable inclusion of bacterial mixture: control, T1, T2, T3, and T4 containing 0, 1.0, 2.0, 3.0, and 4.0 g bacterial mixture/kg diet, respectively. Fish fed the T4 diet had the highest final body weight, weight gain, specific growth rate, thermal growth coefficient, feed intake, and the best feed conversion ratio. No significant differences in body chemical composition of fish were observed in terms of dry matter, crude protein, lipid, ash, and energy content among all treatments. Probiotic supplementation showed no noticeable differences on haematological and biochemical blood parameters, except for packed cell volume, albumin, albumin/globulin ratio, aspartate transaminase, glucose, uric acid, triglycerides, and cholesterol. Conclusively, Nile tilapia fingerlings diet supplemented with evaluated probiotic with an optimal dietary supplementation of 4 g/kg diet improved growth performance and fish health without a negative impact on water quality. This supplementation could therefore be used as a probiotic in Nile tilapia fingerlings diet.