The potential pathogenicity of Beauveria bassiana was examined by intramuscular injection of high (2 x lo8) or low (2 x lo5) concentrations of conidia spores, into the left or right quadriceps muscles of CD-1 mice, respectively. The injection sites were monitored over a period of 28 days by both microbiological and histopathological methods. Focal muscle necrosis, edema and inflammation occurred rapidly (within 12 hours) at the high dose application (2 x 108) site, but such lesions were far less severe with the low dose spore application (2 x 105). Fungal spores in the high dose site persisted in normal shape for 2 weeks, after which time they began to degenerate. Almost all spores were cleared from the injection site within the 28-day observation period. Spread to other organs of the body was not observed, except by macrophage transport to regional lymph nodes. At the low dose rate, most spores were cleared within 12 h to 2 d, leaving only mild focal edema and inflammation. Viable fungal colonies could be recovered up to 3 d after injection from the high dose site, but only up to 12 h from the low dose site. It was concluded that B. bassiana does not cause infection, nor multiply, nor survive for more than 3 days when injected intramuscularly into healthy mice.
B. bassiana belongs to the family of Deuteromycetes (Fungi imperfecti).There is some confusion in the literature, however, over the taxonomic placement of this fungus.
The genomic DNA of two strains of the entomopathogenic fungus Beauveria bassiana, strain GK2016, a "wild type" (virulent), and strain GK2051, a less virulent mutant to grasshoppers, was digested with 12 restriction endonucleases. Gel electrophoresis conditions were established to show restriction fragment length patterns visually in the digested DNA stained with ethidium bromide. The less virulent mutant was generated by ultraviolet illumination of conidiospores at a 95% lethal dose. Both strains of the fungi were identical in morphology as well as in 16 of 22 API-ZYM kit enzyme assays. Differences in levels of total enzyme activity were observed for esterase, esterase-lipase, beta-galactosidase, chitinase, and protease. A Neurospora crassa beta-tubulin gene (heterologous gene) and two homologous DNA probes (pJK16 and pJK18) hybridized to several specific DNA bands in B. bassiana strain GK2016 but not in strain GK2051. Strain GK2051 gave different restriction fragment length pattern when compared with its parent strain. Taken together, the data show restriction fragment length differences between the genomic DNA of the two strains, including the loss of some DNA sequences from the mutant strain, which may be involved in pathogenicity. Finally, B. bassiana GK2016 contains a beta-tubulin gene with at least partial homology to that of N. crassa.
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