Persistence and clearance of the vaccinal strains, humoral and cell-mediated immune response and protective immunity were assessed in ICR mice, immunized intraperitoneally or intragastrically with double-marker auxotrophic mutants of Salmonella typhimurium strains 1771 and 3334 (his-pur-). Strain 3334 possesses in addition the antiepidemic Hst (high sensitivity to tensides) marker which confers decreased survival in the gut and environment. The results showed that the strains are not overattenuated. They revealed preserved multiplication capacity, established carrier state for 28-30 days and induced humoral and cell-mediated immunity as measured by passive haemolysis and microagglutination or footpad swelling tests, respectively. The immune response was determined by biological features of the strains. Strain 1771 induced synthesis of O- and H-antibodies 7-11 days earlier than strain 3334. Footpad reactions appeared 3-4 days after immunization and a week before the serum antibodies. Cell-mediated immunity showed clear correlation with the protection against challenge with a virulent strain. The present results suggest that the double-marker auxotrophic mutants of S. typhimurium might prove very useful in the development of a live vaccine in further studies on livestock.