Sabouraud dextrose chloramphenicol agar

The presence of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) was detected by real-time polymerase chain reactions (genesig^R Real-Time PCR COVID-19, Primerdesign, York, UK) using combined nasal and pharyngeal swabs.
All patients were screened for pulmonary aspergillosis by routinely performing culture and galactomannan analysis on bronchoalveolar lavage fluid (BALF) at least twice weekly. In addition, clotted blood was tested for GM and Beta D-glucan. Each BALF sample was inoculated onto Sabouraud dextrose chloramphenicol agar (Biomerieux, culture media) and incubated at 37 °C for two days and at 30 °C for another five days. Aspergillus spp. were identified by culture characteristics.
Galactomannan analysis on BALF and the serum was performed by sandwich enzyme-linked immunosorbent assay (Platelia^TM Bio-Rad, Watford, UK) as per the manufacturer’s instruction. The mycological criteria for aspergillosis were based on either the growth of Aspergillus species or a BALF galactomannan index (GMI) ≥1.0 or serum GMI ≥ 0.5. (1–3)-β-D-glucan (BDG) was detected using the Fungitell^® assay (Associates of Cape Cod, Liverpool, UK), using a cutoff of 80 pg/mL.

Indoor air was monitored continuously by HTST placed in the waiting room of the transplant unit (HTST-I, unprotected internal air), at 80 cm high, with the same settings as the outdoor HTSTs (n = 28 days) (Fig 1). The first conversion factor used was 25.7 (ie. 0.27% of the surface sampled was read). An extended area of some samples was read as a second step, with a conversion factor of 0.19 (ie. 36.5% of the surface sampled was read). It was also monitored with a viable impaction air sampler device known as a biocollector (AirIDEAL, BioMérieux, Marcy l’Étoile, France), in the same waiting room, at 1 meter high. Paired indoor air samples were collected at the same place twice daily (at 10 a.m. and 2 p.m. ± 2h), from Monday to Friday, excluding public holidays (n = 19 days). The biocollector draws in air at 100 L/min for 1 min. Overall, 100 L were collected on Sabouraud Dextrose Chloramphenicol agar (BioMérieux) and incubated for 5–7 days at 30°C for the 1^st plate and for 48 hours at 37°C for the 2^nd plate. Viable fungal colonies (colony forming units) were counted and identified. TFLs and Aspergillus spp. loads were expressed as colony-forming units (CFU)/m³.

Sputum sample was inoculated onto Sabouraud Dextrose Chloramphenicol agar (BIOMERIEUX, Culture media, Basingstoke, UK) and incubated at 37 °C for two days and at 30 °C for another five days, except sputum samples from Cystic Fibrosis (CF). CF cultures were incubated at 30 °C for four weeks. Aspergillus spp. were identified by culture characteristics supplemented with Matrix-Assisted Laser Desorption/Ionisation-Time of Flight (Bruker, MALDI-TOF, MS, Coventry, UK).