Reduced susceptibility to fluconazole impairs <i>C. gattii</i> migration to the CNS.

<p>Lungs (<b>A</b>), brain (<b>B</b>), and Bronchoalveolar lavage fluid (BALF) (<b>C</b>) 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<b>_F</b> strains. ND: non-detected. Black bars refer to L27/01 and grey bars refer to L27/01<b>_F</b>. *Data represent the mean of two independent experiments in triplicate. P<0.05 was considered to be significant. (<b>D–H</b>) Histopathological sections stained with H&E of lung and brain 15 days post-inoculation. Lungs of control mice (<b>D</b>). 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 (<b>E</b>). Lungs of mice inoculated with L27/01<b>_F</b> strain, with intense diffuse inflammatory infiltrate, neutrophils and macrophages around few yeast cells (head arrow) (<b>F</b>). Bar = 40 µm. Hippocampi of mice inoculated with L27/01 strain showed cryptococcoma (arrow), neuronal degeneration and death (head arrow). <i>Cryptococcus gattii</i> yeasts in cryptococcoma were stained with Groccot’s stain (in detail) (<b>G</b>). Hippocampi of mice inoculated with L27/01<b>_F</b> strain showed unaltered structure (<b>H</b>). Bar = 20 µm.