In this study, the thirty-two investigated Propionibacterium spp. strains were examined for their abilities to survive at pH 2.0, 3.0, 4.0 and 5.0 and in the presence of 0.06, 0.15 and 0.30 % bile salts, their influence on the growth of pathogens, as well as their sensitivity against eleven selected antibiotics. P. freudenreichii subsp. freudenreichii IT4, IT16, and P. jensenii MP12, BSM8 and DY1 strains survived in both the acidic and bile salt environments. Among thirty-two tested Propionibacterium spp. strains, five strains demonstrated strong and broad inhibitory spectrum against pathogen strains, and all of the Propionibacterium spp. strains strongly inhibited growth of the Escherichia coli ATCC 11229. Propionibacterium spp. strains demonstrated resistance to streptomycin, gentamycin, kanamycin, rifampicin and polymyxin B antibiotics. Overall, Propionibacterium spp. strains showed high antibacterial activity, tolerance to pH 4.0 and 5.0 and high tolerance to bile salts and will present an alternative source to Lactobacillus and Bifidobacterium for future probiotic development. In addition, culture metabolites of these strains may be used as a preservative in the food industry for the replacement of or reduction in the increasing use of chemical additives.