Abstract: The present study was aimed to isolate, characterize and examine the probiotic properties of Bacillus infantis (KADR2) from Labeo rohita. The total of seven strains isolated from gastrointestinal tract of Labeo rohita, one of them KADR2 showed higher antagonistic effect against fish pathogens. The strain was evaluated under in vitro intestinal condition based on resistance to bile tolerance, low pH, hydrophobicity, catalase activity and antibiotics susceptibilities. Partial 16S rRNA gene sequence of this strain KADR2 were blasted and showed homology with Bacillus infantis(99%) supported by morphological and physiological characterization. Consequently, the positive results of this study suggested that further studies in challenge experiments in fish to explore their probiotic effects having great scope for being used as a potential probiotic in aquaculture.