Sterile falcon tubes

Both respiratory tract specimens (BAL, BL, sputum) and body fluids (peritoneal, pleural and CSF) were collected in 50 ml sterile Falcon tubes (Becton Dickinson) and centrifuged at 1550 × g for 30 min (Centra MP4R, IEC). Fresh tissues (biopsy) were manually homogenized in 3 ml of sterile saline solution. The pelleted samples and the homogenate obtained were used for culture and stain, and 0.6 ml of each sample was stored at −20° C for subsequent DNA extraction.
Processed specimens were cultured on Sabouraud Dextrose Agar and Mycosel (Becton Dickinson), incubated at room temperature (±18–22° C) for three weeks, and examined weekly for yeast colonies; the identification was performed using microscopic observation, phenotypic type enzyme (urease and phenoloxidase), and carbohydrate assimilation patterns (McTaggart et al., 2011).¹⁴ (link) The biovarieties were determined by culturing the isolates on l-canavanine glycine bromothymol blue (CGB) selective medium (Klein et al., 2009).³² (link) These procedures were carried out in a Biosafety Level 3 (BSL3) laboratory.

Respiratory tract specimens (BAL, BL fluid and sputum) and body fluids, including peritoneal fluid, pleural fluid and cerebrospinal fluids (CSF) were collected in 50 mL sterile Falcon tubes (Becton Dickinson) and centrifuged at 1550 × g for 30 min (Centra MP4R, IEC). The fresh tissues (biopsies) were manually homogenized in 3 mL of sterile saline solution. The pelleted samples and the homogenate obtained were used for culture and Gomori methenamine stain, and 0.6 mL of each sample was stored at −20 °C for subsequent DNA extraction. Processed clinical samples were cultured on Sabouraud dextrose agar and Mycosel (Becton Dickinson) by incubation at room temperature (18–22 °C) for 20–30 days and were examined weekly macroscopically and microscopically. These procedures were carried out in a biosafety laboratory level 3 (BSL3).