Influences of the Culturing Media in the Virulence and Cell Wall of Sporothrix schenckii, Sporothrix brasiliensis, and Sporothrix globosa

Abstract

<i>Sporothrix schenckii</i>, <i>Sporothrix brasiliensis</i>, and <i>Sporothrix globosa</i> are etiological agents of sporotrichosis, a human subcutaneous mycosis. Although the protocols to evaluate <i>Sporothrix</i> virulence in animal models are well described, the cell preparation before inoculation is not standardized, and several culturing media are used to grow yeast-like cells. Here, we found that carbon or nitrogen limitation during fungal cell preparation negatively impacted the ability of <i>S. schenckii</i> and <i>S. brasiliensis</i> to kill <i>Galleria mellonella</i> larvae, but not <i>S. globosa</i>. The fungal growth conditions associated with the short median survival of animals were accompanied by increased hemocyte countings, phenoloxidase activity, and cytotoxicity. The fungal growth under carbon or nitrogen limitation also affected the cell wall composition of both <i>S. schenckii</i> and <i>S. brasiliensis</i> and showed increased exposure of β-1,3-glucan at the cell surface, while those growing conditions had a minimal impact on the <i>S.</i><i>globosa</i> wall, which had higher levels of this polysaccharide exposed on the wall regardless of the culture condition. This polysaccharide exposure was linked to the increased ability of insect hemocytes to uptake fungal cells, suggesting that this is one of the mechanisms behind the lower virulence of <i>S.</i><i>globosa</i> or cells from the other species grown in carbon or nitrogen limitation.