Three strains of Lactobacillus casei and one of Lactobacillus helveticus were examined for antagonistic activity toward twenty five indicator strains of different species. Under conditions eliminating the effects of organic acid and hydrogen peroxide, culture supernatants of all the Lactobacillus strains exhibited a wide spectrum of inhibitory activity toward microorganisms of different genera. The inhibitory compound secreted by one strain of L. casei was active against Clostridium tyrobutyricum. The active components were insensitive to proteolytic enzyme and heat treatment.