DESCRIPTIONS OF MEDICAL FUNGI

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36 Descriptions of Medical Fungi Candida Berkhout Dalmau Plate Culture: To set up a yeast morphology plate, dip a flamed sterilised straight wire into a culture and then lightly scratch the wire onto the surface of a cornmeal/tween 80, rice/tween 80 or yeast morphology agar plate, then place a flamed coverslip onto the agar surface covering the scratches. Dalmau morphology plates are examined in-situ directly under the low power of a microscope for the presence of pseudohyphae which may take up to 4-5 days at 26 O C to develop. C. albicans also produces characteristic large, round, terminal, thick-walled vesicles (often called chlamydospores). For best results a light inoculum should be scratched into the agar surface using a wire. 10 μm a 5 mm b Dalmau plate culture of Candida albicans showing (a) colonies growing out from scratches on the surface of a cornmeal/tween 80 agar plate, and (b) the production of large round, thick-walled chlamydospores. Note: A coverslip has been placed onto the agar surface covering the scratches. Physiological and Biochemical Tests: including fermentation and assimilation studies should be performed based on those used at the Centraalbureau voor Schimmelcultures (CBS), Delft, The Netherlands (Kurtzman et al. 2011). Reliable commercially available yeast identification systems are the API 20C AUX, API ID 32C, Biolog YT Station and Vitek 2 YST ID systems. However, they can only be used to identify those species in their respective databases, and may misidentify yeasts that are not represented. Other Supplementary Tests include growth at 37 O C, cycloheximide resistance and hydrolysis of urea. MALDI-TOF MS: The Bruker MALDI-TOF database is useful for identification of most clinical yeasts. The MALDI-TOF Vitek MS has been reported to misidentify some yeasts, notably Candida metapsilosis as Candida parapsilosis (Nobrega et al. 2014). Molecular Identification: ITS sequencing is useful for the identification of most clinical yeasts. References: Barnett et al. (1983), Kurtzman and Fell (1998, 2011), de Hoog et al. (2000, 2015).
Descriptions of Medical Fungi 37 Candida albicans is a commensal of mucous membranes and the gastrointestinal tract. Environmental isolations have been made from sources contaminated by human or animal excreta, such as polluted water, soil, air and plants. RG-2 organism. Candida albicans (Robin) Berkhout Culture: Colonies (SDA) white to cream-coloured smooth, glabrous, yeast-like. Microscopy: Spherical to subspherical budding blastoconidia, 2-7 x 3-8 µm in size. India Ink Preparation: Negative - no capsules present. Dalmau Plate Culture: Branched pseudohyphae with dense verticils of blastoconidia. Spherical chlamydospores, mostly terminal, often on a slightly swollen subtending cell, are formed near the edge of the cover slip. Physiological Tests: + Positive, - Negative, v Variable, w Weak, s Slow Germ Tube + L-Sorbose v L-Arabinose v D-Glucitol v Fermentation Sucrose v D-Arabinose v α-M-D-glucoside v Glucose + Maltose + D-Ribose v D-Gluconate v Galactose v Cellobiose - L-Rhamnose - DL-Lactate + Sucrose v Trehalose v D-Glucosamine v myo-Inositol - Maltose + Lactose - N-A-D-glucosamine v 2-K-D-gluconate + Lactose - Melibiose - Glycerol v D-Glucuronate - Trehalose v Raffinose - Erythritol - Nitrate - Assimilation Melezitose v Ribitol v Urease - Glucose + Soluble Starch + Galactitol - 0.1% Cycloheximide + Galactose + D-Xylose + D-Mannitol + Growth at 40 O C + Key Features: Germ tube positive, production of chlamydospores on Dalmau plate culture, fermentation of glucose, sugar assimilation profile and a distinctive green colour on CHROMagar. Note: Germ tube negative variants (previously known as C. claussenii), and sucrose-negative variants (previously described as C. stellatoidea) may occur. Antifungal Susceptibility: C. albicans (Australian National data); MIC µg/mL. CLSI clinical breakpoints are marked where available (Pfaller and Diekema 2012). No 64 AmB 1725 2 16 162 644 548 305 48 FLU 1728 2 2 2 83 468 706 314 62 12 11 18 8 40 VORI 1445 739 404 156 75 18 17 18 5 5 1 1 6 POSA 1095 88 416 367 137 50 18 10 6 1 1 ITRA 1728 20 122 443 661 378 43 30 10 3 1 17 ANID 821 5 280 320 146 64 5 0 0 1 MICA 819 470 261 73 11 4 CAS 1171 3 13 207 490 327 112 17 2 5FC 1728 4 147 765 362 164 171 62 16 7 5 3 3 19
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DESCRIPTIONS OF MEDICAL FUNGI

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