The enteric flora comprise approximately 95% of the total number of cells and are capable of eliciting immune responses while also protecting against microbial pathogens. Probiotic microorganism shows bile tolerance, acid resistance and antibiotic resistance. Isolated probiotic microorganism from stool, curd, milk sample shows better activity as compare to commercially available probiotics. Natural samples were having bacilli, cocci and yeast while commercial samples were having bacilli and and cocci. The isolates from natural samples proved to be having higher antimicrobial activity against selected pathogens as compared to commercially available probiotics. These microorganisms can be used in therapeutics for the treatment of gastro-intestinal diseases and can be administered with several antibiotics.