Aquaculture industry is the fastest-growing food producing sector but fish diseases has been a major threat to the growth. Bacterial infections are one of the causes of mortality in fish and the use of antibiotics has resulted in resistant strain of bacteria in fish. Therefore role of probiotics is being considered as alternative. Studies were carried out to isolate and identify probiotics from the gut of Tilapia guineensis. Guts removed from dissected fish were suspended in 9ml of saline diluents homogenized and allowed to pour in MRS broth and agar medium. The isolated lactobacillus was re dissolved and shaken homogenously incubated at 370C aenarobically for 24 hrs. The culture obtained were subculture on MRS at pH 6.2 for discrete colonies. Single colony of lactobacillus was isolated using colony morphology and some biochemical tests. The isolated bacteria were identified using biochemical test and API CH in conjunction with API CHL medium for the identification of probiotics. Bacteria isolated from the gut of Tilapia guineensis were identified as pediococcus pentosaceus2 and pediococcus pentosaceus1 by observing their colony morphology, physiological and as well as some biochemical characteristics. Microscopically they were Gram-positive, rod shaped, non- motile, catalase negative and absence of Endospore.