Strains used for this study have been verified by rDNA internal transcribed spacer (ITS) and partial β-tubulin (BT2) sequencing and compared with ex-type isolates in the reference collection of the CBS-KNAW Fungal Biodiversity Centre, Utrecht, the Netherlands. We analysed 32 strains of Pseudallescheria, Petriellopsis and Scedosporium (Table 1). Methods of DNA extraction, alignment and phylogenetic analysis were those of Badali et al. [18] Species attribution was verified by sequencing ITS rDNA and partial β-tubulin (BT2) according to Gilgado et al. [10] and by comparing them with ex-type isolates from the reference collection of CBS (Utrecht, the

Netherlands). Pseudallescheria angusta and P. ellipsoidea https://www.selleckchem.com/products/PD-0325901.html are listed as part of P. boydii. Three different microtitre plates were used with the Taxa Profile Micronaut system (Merlin Diagnostika GmbH): Taxa Profiles A, C and E. On each microtitre plate, two strains were analysed synchronously for 191 reactions in the case of Taxa Profiles A and C (one growth control) and 188 reactions for Taxa Profile E (three negative controls and one growth control). Taxa Profile A contains amines, amides, amino acids, other organic acids, and includes heterocyclic aromatic compounds. Taxa Profile C contains mono-, di-, tri- and polysaccharides, and sugar derivatives.

On panels A and C, each well contains 1.6 g L−1 of the respective chemical compound. Results were read Selleck Romidepsin visually and photometrically at 620 nm (single scan). Taxa Profile E contains 95 aminopeptidase and protease reactions, 76 glycosidases, phosphatidases and esterases (each for testing at the different pH values of 8.2, 7.5, 5.5 and 4.0), desaminases and decarboxylases (arginine-dihydrolase, glutamate-, lysine-, ornithine-decarboxylases and relevant control reactions), and 17 classical reactions (such as urease, indol, nitrate and nitride). A full

list of the reactions is provided in Supporting Information. Strains were cultured on potato dextrose agar (PDA; Oxoid, Wesel, Germany), Sabouraud’s 4% glucose agar, water agar, Müller Hinton’s agar and Columbia sheep blood agar. The incubation period was up to 7 days at 35 ± 1 °C to Immune system obtain optimal conidiation. The plates were covered with 5–6 ml sterilised 0.9% NaCl solution. Conidia were scraped off carefully and transferred into a sterile glass tube using a sterile pipette. The suspensions were vortexed and centrifuged for 5 min at 21 °C at 3000 rpm; sediments were washed three times in 5 ml sterile 0.9% NaCl solution. Suspensions were adjusted with a UV 160 spectrophotometer for Taxa Profiles A and C panels to 0.150–0.170 at 530 nm (1–5 × 104 colony forming units ml−1),19 and to 0.20–0.28 at 560 nm for Taxa Profile E.