Francisella tularensis gives rise to two distinct colony types, acriflavine agglutination test-positive (acf+) and -negative (acf-) colonies. The acf+ variants were exclusively low virulent in mice, while the acfvariants were shown to be either high or low virulent.Three fractions, phosphate-buffered salineextractable without heating, with heating at 60 C, and with heating at 100 C, were obtained from cultures of both the acf+ and acf-variants on agar media, and the polysaccharide antigens in those fractions were quantitated.All of the highly virulent acf-variants possessed a large amount of the polysaccharide antigen in the fraction extractable with heating at 60 C. This antigen was not, however, detected in any of the acf+ variants and one low-virulent acf-variant. It was also detected in a very low amount in some other acf-variants with low virulence. The amount of this polysaccharide antigen was therefore shown to be correlated with bacterial virulence in mice.