Spontaneous mutant of Blastomyces dermatitidis attenuated in virulence for mice

A spontaneous mutation occurred in cultures of a virulent strain of Blastomyces dermatitidis during serial passage at 35-37 degrees C. The mutant strain was shown to be 10,000-fold less virulent for mice than the parent strain, and this was independent of the challenge dose, the age of the challenged host, and the route of challenge (although attenuation was greater for pulmonary than for peritoneal challenge). The mutant grows as well as the parent in fungal media or in the presence of murine substrates. Studies of the chronology of pulmonary infection indicate the mutation affects an early event (first week after challenge) in establishment of infection and interaction with host defenses. Scanning electron microscopy showed that the mutant lacks deep irregular surface ridges and grooves present in the parent. It had increased resistance to crystal violet in vitro, increased susceptibility to bile, and could hydrolyze esculin. The latter findings are of interest because they place the mutant in an intermediate position with respect to these properties between the virulent parent and a previously studied avirulent strain. Lipid analyses indicated a marked increase in fatty acids in the mutant. Studies of four B. dermatitidis strains have also associated increasing palmitic acid and phospholipid content with increasing virulence.