The antagonistic activity of probiotic strains (Bifidobacterium animalis BB-12, Bifidobacterium bifidum A, Bifidobacterium longum B6, Lactobacillus acidophilus ADH, Lactobacillus paracasei ATCC 25598, and Lactobacillus rhamnosus GG) against nalidixic acid resistant ($NA^R$) Escherichia coli O157:H7 MF1847, E. coli O157:H7 H2439, E. coli O157:H7 ATCC 43894, and E. coli O157:H7 C7927 was investigated using the agar-overlay, well diffusion, and broth culture tests. L. paracasei ATCC 25598 was the most effective probiotic strain in terms of in vitro antagonistic activity against $NA^R$ E. coli O157:H7, followed by L. rhamnosus GG, B. longum B6, and L. acidophilus ADH. The use of selected probiotic strains could be an effective pre-harvest intervention strategy to reduce the risk of $NA^R$ E. coli O157:H7 by maintaining a balanced microflora in animals and might provide many potential benefits in lieu of using antimicrobials.