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Cryptococcus gattii is responsible for an expanding epidemic of serious infections in Western Canada and the Northwestern United States (Pacific Northwest). Some patients with these infections respond poorly to azole antifungals, and high azole MICs have been reported in Pacific NorthwestC. gattii . In this study, multiple azoles (but not amphotericin B) had higher MICs for 25 Pacific NorthwestC. gattii than for 34 non-Pacific NorthwestC. gattii or 20Cryptococcus neoformans strains. We therefore examined the roles in azole resistance of overexpression of or mutations in the gene (ERG11 ) encoding the azole target enzyme.ERG11 /ACT1 mRNA ratios were higher inC. gattii than inC. neoformans , but these ratios did not differ in Pacific Northwest and non-Pacific NorthwestC. gattii strains, nor did they correlate with fluconazole MICs within any group. Three Pacific NorthwestC. gattii strains with low azole MICs and 2 with high azole MICs had deduced Erg11p sequences that differed at one or more positions from that of the fully sequenced Pacific NorthwestC. gattii strain R265. However, the azole MICs for conditionalSaccharomyces cerevisiae erg11 mutants expressing the 5 variantERG11 s were within 2-fold of the azole MICs forS. cerevisiae expressing theERG11 gene fromC. gattii R265, non-Pacific NorthwestC. gattii strain WM276, orC. neoformans strains H99 or JEC21. We conclude that neitherERG11 overexpression nor variations inERG11 coding sequences was responsible for the high azole MICs observed for the Pacific NorthwestC. gattii strains we studied.

Azole Resistance in Cryptococcus gattii from the Pacific Northwest: Investigation of the Role of ERG11