Phenotypic switching of Candida guilliermondii is associated with pseudohyphae formation and antifungal resistance.

Abstract

Switching between two cell types in fungi is called phenotypic switching, and it is commonly observed in pathogenic yeast. Candida lusitaniae undergoes antifungal resistance-associated phenotypic switching and results in three colony colors: light brown, brown and dark brown. In this study, we included C. lusitaniae as control. This study had two objectives. First, we wanted to evaluate whether also a prevalent human pathogen C. guilliermondii can undergo phenotypic switching. Second, our aim was to determine whether switching can change yeasts susceptibility to antifungals. Yeast suspension (1 × 10(3)-5 × 10(3) c.f.u./ml) was plated on the YPD medium containing 1 mM CuSO4. Colonies exhibiting the original and variant phenotypes were counted and converted to percentage of the population. Minimum inhibitory concentrations of amphotericin B, formic acid and acetic acid for the cells from random colonies of the different phenotypes were determined by microdilution method. After 5 days of incubation, C. guilliermondii switched spontaneously and reversibly among two phenotypes distinguishable on CuSO4 containing agar, white and dark brown. Phenotypes occurred with greater frequency (10(-1)-10(-2)) than spontaneous mutations and were reversible, fulfilling the two phenotypic switching criteria. The study showed that phenotypic switching was associated with filamentation and affected antifungal resistance. Resistance to amphotericin B increased tenfold and was associated with C. lusitaniae dark brown phenotype. C. guilliermondii colonies with brown phenotype displayed 20 and 2 times higher resistance to amphotericin B and acetic acid, respectively.