Stereo discovery v20 dissecting microscope

Species were characterised using standard growth conditions (Okuda et al. 2000, Visagie et al. 2013, Visagie et al. 2014a ). Strains were inoculated in three-point fashion onto CYA, MEA, Yeast Extract Sucrose agar (YES), DG18, CYA with 5 % NaCl (CYAS), Oatmeal agar (OA) and Creatine Sucrose agar (CREA). Plates were incubated in plastic boxes for 7 d in the dark at 25 °C. Additional CYA plates were incubated at 30 and 37 °C. Colour names and alphanumeric codes used in descriptions refer to Kornerup & Wanscher (1967) .
Microscopic preparations were made from colonies grown on MEA, with DG18 also used for Aspergillus, after 1 to 2 wk. Lactic acid (60 %) was used as mounting fluid and excess conidia were washed away with 70 % ethanol. Characters were captured using a Zeiss SteREO Discovery.V20 dissecting microscope and Zeiss AX10 Imager.A2 compound microscope, both equipped with AxioCam MRc5 cameras using AxioVs40 v. 4.8.2.0. Microscopic measurements were done using Nikon NIS-elements D v. 4.0. Photo plates were prepared in Adobe^® Photoshop^® CS6 with photomicrographs cleaned up using the healing brush tool, for aesthetic reasons, without altering areas of scientific significance.

Macroscopic characters were studied on Czapek yeast autolysate agar (CYA), CYA supplemented with 5 % NaCl (CYAS), yeast extract sucrose agar (YES), creatine sucrose agar (CREA), dichloran 18 % glycerol agar (DG18), oatmeal agar (OA) and malt extract agar (MEA; Oxoid malt) (Samson et al. 2010 ). Isolates were inoculated at three points on 90 mm Petri dishes and incubated for 7 d at 25 °C in darkness. Additional CYA plates were incubated at 30 and 37 °C, while additional MEA plates were incubated at 30 °C. After 7 d of incubation, colony diameters were recorded. The colony texture, degree of sporulation, obverse and reverse colony colours, the production of soluble pigments and exudates were noted. Acid production on CREA is indicated by a change in the pH sensitive bromocresol purple dye, from a purple to yellow colour in media surrounding colonies. For ascoma production, OA, MEA and CYA plates were incubated for up to four weeks.
Microscope preparations were made from 1 wk old colonies grown on MEA and ascomata, asci and ascospores were observed on OA. Lactic acid (60 %) was used as mounting fluid and 96 % ethanol was applied to remove the excess of conidia. A Zeiss Stereo Discovery V20 dissecting microscope and Zeiss AX10 Imager A2 light microscope equipped with Nikon DS-Ri2 cameras and software NIS-Elements D v4.50 were used to capture digital images.

Macroscopic characters were studied on Czapek Yeast Autolysate agar (CYA), CYA supplemented with 5 % NaCl (CYAS), Yeast Extract Sucrose agar (YES), Creatine Sucrose agar (CREA), Dichloran 18 % Glycerol agar (DG 18), Oatmeal agar (OA) and Malt Extract agar (MEA, Oxoid malt) (Samson et al. 2010 ). To enhance the growth, the ex-type culture of A. acidohumus CBS 141577, isolated from acid soil from China, was additionally inoculated on Cherry Decoction agar (CHA) (Crous et al. 2009 ). The isolates were inoculated at three points on 90 mm Petri dishes and incubated for 7 d at 25 °C in darkness. In addition, CYA and MEA plates were incubated at 30 °C and 37 °C. After 7 d of incubation, colony diameters were recorded. The colony texture, degree of sporulation, obverse and reverse colony colours, the production of soluble pigments and exudates were determined. Light microscope preparations were made from 1 wk old colonies grown on MEA. Lactic acid (60 %) was used as mounting fluid. Ethanol (96 %) was used to remove excess conidia and prevent air bubbles. A Zeiss Stereo Discovery V20 dissecting microscope and Zeiss AX10 Imager A2 light microscope equipped with Nikon DS-Ri2 cameras and software NIS-Elements D v4.50 were used to capture digital images.