Bacitracin and neomycin have been used for a long time for local intestinal antisepsis or decontamination, due to their scarce or nonexistent intestinal absorption. The aim of the present study was to determine the activity of bacitracin and neomycin against recent clinical isolates of aerobic and anaerobic microorganisms and to verify their capacity to have a synergistic effect and to prevent the emergence of bacterial resistance when in combination. The results showed that the activity of either antibiotic against recent clinical isolates (even if resistant to cephalosporins and aminoglycosides) is similar to that originally displayed at the time introduction in therapy. Generally the combination of the two antibiotics showed a synergistic or additive effect and prevented the selection of resistant strains.