Abstract: The objectives of this study were to isolate, select and identify lactic acid bacteria (LAB) for the probiotic properties in cattle. Small, large intestines and feces samples were collected from 6 healthy Native x Brahman cattle from the animal farm division of Faculty of Agriculture, Khon Kaen University, Thailand. All samples were cultivated using a modified De Man, Rogosa and Sharp (MRS) agar supplemented with 0.3% CaCO3 (w/v). Bacterial colonies which showed clear zone surrounding their colonies were selected to a further test of the basic probiotic properties including acid and bile tolerance test, anti-pathogenic bacteria. Safety features such as antimicrobial susceptibility were also tested using the disk diffusion method. The results showed that the concentration of LAB from small intestine, large intestine and feces were 5.15 x 107, 5.85 x 107 and 1.25 x 1012 CFU/g, respectively. Twenty seven out of 86 isolates tolerated to pH 3 and 15 isolates tolerated to bile salt. Fifteen out of these acid-and bile-tolerant isolates showed the ability to inhibit Escherichia coli ATCC 25923 and Salmonella Typhimurium. All acid and bile tolerant isolates were highly sensitive to penicillin, erythromycin, tetracycline and vancomycin. In contrast, 11 and 4 isolates resisted to streptomycin and gentamicin, respectively. However, only one isolate (F30) resulted in resisting to low pH, bile salt and anti-pathogenic bacteria. This isolate was identified using 16S rRNA gene sequencing as Streptococcus sp., [closely related to Streptococcus infantarius (99.93%)] and shown as a potential probiotic in cattle.