Figure 5. Reduced susceptibility to fluconazole impairs C. gattii migration to the CNS.

Lungs (A), brain (B), and Bronchoalveolar lavage fluid (BALF) (C) were removed (n = 8), homogenized, diluted and plated onto Sabouraud dextrose agar for measurement of fungal burden 0.5, 1, 7, 15, and 30 days post-infection with 10⁶ cells of L27/01 or L27/01_F strains. ND: non-detected. Black bars refer to L27/01 and grey bars refer to L27/01_F. *Data represent the mean of two independent experiments in triplicate. P<0.05 was considered to be significant. (D–H) Histopathological sections stained with H&E of lung and brain 15 days post-inoculation. Lungs of control mice (D). Lungs of mice inoculated with L27/01 strain showing mild peribronchiolar and moderate perivascular neutrophilic and histiocytic inflammatory infiltrate associated with numerous yeast cells in the peribronchiolar and alveolar lumen (E). Lungs of mice inoculated with L27/01_F strain, with intense diffuse inflammatory infiltrate, neutrophils and macrophages around few yeast cells (head arrow) (F). Bar = 40 µm. Hippocampi of mice inoculated with L27/01 strain showed cryptococcoma (arrow), neuronal degeneration and death (head arrow). Cryptococcus gattii yeasts in cryptococcoma were stained with Groccot’s stain (in detail) (G). Hippocampi of mice inoculated with L27/01_F strain showed unaltered structure (H). Bar = 20 µm.