Experiments were undertaken to determine whether female leghorn chickens that had been mated had a lower incidence of salmonellae colonization in the ovary, oviduct, internal organs, and eggs than did unmated birds from the same flock when both were inseminated with Salmonella enteritidis-contaminated semen. Eggs were sampled at approximate 3-day intervals, and fecal samples were taken at 24 hours and 7 days after insemination. Fourteen days following insemination, hens were euthanatized and the ovary, oviduct, liver, spleen, gall bladder, and ceca were removed. None of the previously mated hens had salmonellae present at any sampling time. Virgin birds showed salmonellae colonization in all samples. This indicates that mating activity and the environment of females offers some protection against salmonellae colonization in the reproductive tract.