One-hundred-and-five strains of Salmonella enterica serovar gallinarum, biovar pullorum and 43 strains of biovar gallinarum were characterized by biochemical reactions, and 95 strains of biovar pullorum and 32 strains of biovar gallinarum were examined for plasmid content. Twenty-one (66%) of the strains classified as biovar gallinarum contained a 85-kb virulence plasmid. Six of these strains also contained a 2.5-kb plasmid. Ten strains (31%) were without plasmids. Ninety-three strains (98%) belonging to biovar pullorum contained the virulence plasmid and, in addition, these strains contained from one to four small plasmids. Two strains did not contain plasmids. Restriction enzyme digestions of the virulence plasmid showed three different profiles, two of which were common for biovars gallinarum and pullorum. It is suggested that the phenotypic diversity of biovars gallinarum and pullorum, as well as the diversity of plasmid profiles of biovar pullorum, could be used as epidemiological markers. Differences in phenotypic characters were not related to plasmids demonstrated in biovars of S. enterica serovar gallinarum.