Oral administration to newly hatched chickens or to chicks up to 5 days of age with an avirulent, rough, spectinomycin-resistant mutant of Salmonella typhimurium strain F98 inhibited the colonization of a nalidixic acid-resistant mutant of the same strain administered by the same route 1 day later. The second strain passed rapidly through the alimentary tract and persisted in the caeca of only a few chickens. Resistance to colonization did not develop until 24 h after inoculation of the first strain but was still evident if the second strain was inoculated up to 7 days later. Resistance occurred in 5 different breeds of chicken and in chickens reared on 5 different diets. Protection was evident against a very high challenge dose and could be produced by the introduction of small numbers of the first strain. Pre-colonization of chicks with the first strain of F98 reduced faecal excretion of the second strain over many weeks, whether chickens were challenged directly or by contact with other infected chickens. The rough strain F98 produced protection against only a few S. typhimurium strains and not against other serotypes. However, strains of S. infantis and S. heidelberg, chosen because they colonized the chicken alimentary tract better than did F98, produced inhibition of a wider range of serotypes.