The aim of this study was to investigate the antimicrobial and biopreservation potential of lactic acid bacteria. The potential probiotic culture inhibited the growth of gram-positive and gram-negative foodborne pathogens in agar spot assay with inhibition zones ranging from 10 to 21 mm in diameter. The strains showed coaggregation capabilities ranging from 7 to 71% with tested food pathogens including Escherichia coli, Staphylococcus aureus, Listeria monocytogenes, and Salmonella enterica subsp. enterica serovar Typhimurium. The effect of cell-free supernatants on the release of 260 nm absorbing material, especially nucleic acids, was evaluated and indicated the antagonistic activity on foodborne pathogens, the highest being Lactobacillus paraplantarum against E. coli (3.77) and S. aureus (3.86) after 60 min. The effect of cell-free supernatant (CFS) on the growth of pathogens showed that Lactobacillus paraplantarum 11 and L. pentosus 93 had the highest inhibitory activity against tested strains. The biopreservation assay indicated that the potential probiotic strains Lactobacillus paraplantarum 11 (BT), Lactiplantibacillus plantarum 19, Lactobacillus pentosus 42, Limosilactobacillus fermentum 60, Lactobacillus pentosus 93, and Limosilactobacillus reuteri 112 were effective in reducing the Listeria monocytogenes population in raw buffalo milk. Complete Listeria monocytogenes inhibition was observed after 6-8 days. This study showed that probiotic LAB from buffalo milk have antimicrobial and biopreservation potential; these strains have the potential to be utilized as biopreservative agents in food products.