Fungal Strain Cultivation and Quantification

C. albicans strain SC5314 was used in this study (He et al. 2015 (link)). The standard Candida species involved, including C. tropicalis ATCC1369, C. glabrata ATCC15126, C. parapsilosis ATCC22019 and C. krusei ATCC6258, as well as a standard Saccharomyces cerevisiae strain ATCC9763 were obtained from the American Type Culture Collection. The fungi were cultured to exponential phase at 35 °C, 5% CO₂ in yeast extract peptone dextrose (YPD) medium (1% yeast extract, 2% peptone, 2%D-glucose) and harvested by centrifugation at 400 × g for 5 min. After two washes with PBS, the fungal density was determined using a cell counting plate. A Candida count of 5 × 10⁷ CFUs was used to choose the suitable digestive enzymes; a Candida count of 5 × 10⁵ CFUs was utilized to validate the ideal lysis conditions, which may be closer to the fungal load in real applications.