The present study aimed to isolate and characterize potential probiotic strains from the gut microbiota of Labeo rohita, a widely consumed freshwater fish species. A total of 19 isolates were screened out from the gut of rohu. Out of this, two were evaluated with autochthonous probiotic properties. This strains were able to grow at pH 3 – 8.5 with the highest survival and growth rate in neutral conditions (pH 7) for two isolates. Furthermore, all isolates are tolerated bile salt concentrations of 0, 0.15 and 0.3%. The isolation of autochthonous probiotics from the gut of Labeo rohita highlights the presence of indigenous beneficial microorganisms that could play an important role in maintaining the health and well being of the host fish species.