DESCRIPTIONS OF MEDICAL FUNGI

DESCRIPTIONS OF MEDICAL FUNGI 

SECOND EDITION 

DAVID ELLIS 

STEPHEN DAVIS 

HELEN ALEXIOU 

ROSEMARY HANDKE 

ROBYN BARTLEY 

MYCOLOGY UNIT 

WOMEN’S AND CHILDREN’S HOSPITAL 

SCHOOL OF MOLECULAR & BIOMEDICAL SCIENCE 

UNIVERSITY OF ADELAIDE 

ADELAIDE 

AUSTRALIA 

2007 

Cover: Cryptococcus neoformans, and montages including Microsporum, Candida, 

Schizophyllum, Sordaria, Conidiobolus, Fusarium, Bipolaris, Aspergillus, Curvularia, 

Saksenaea, Gliocladium, Trichophyton and Phialophora.

Published by the Authors 

Mycology Unit 

Women’s and Children’s Hospital 

North Adelaide 5006 

AUSTRALIA 

Direct Phone: (08) 8161 7365 

International + 618 8161 7365 

Direct Fax: (08) 8161 7589 

International + 618 8161 7589 

Email: dellis@adelaide.edu.au 

www.mycology.adelaide.edu.au 

© Copyright 2007 

The National Library of Australia Cataloguing-in-Publication entry: 

Descriptions of medical fungi. 

2nd ed. 

Bibliography. 

Includes index. 

ISBN 9780959851267 (pbk.). 

1. Fungi - Indexes. 2. Mycology - Indexes. I. Ellis, David (David H.). 

579.5 

Printed in Adelaide by 

Nexus Print Solutions 

153 Holbrooks Road 

Underdale, South Australia 2032

The Mycology Unit at the Adelaide Women’s and Children’s Hospital has played a key 

role in the provision of the Mycology component of the Microbiology Quality Assurance 

Program (QAP) organised by the Royal College of Pathologists of Australasia 

since its inception in 1979. The idea to provide all laboratories with a set of description 

sheets covering medical fungi evolved in the late 1980s and the first edition of this 

book was published in 1992. We now provide an updated edition which includes new 

and revised descriptions. We have endeavoured to reconcile current morphological 

descriptions with more recent genetic data, however in some cases, especially for the 

anthropophilic dermatophytes this is currently not possible. 

These descriptions have by necessity been kept brief and many have been based on 

previous descriptions by other authors. For further information regarding any of the 

mycoses or pathogenic fungi mentioned, the reader is referred to the references cited. 

For the precise definitions of the mycological terminology used, the reader is referred 

to Ainsworth and Bisby’s Dictionary of the Fungi (Kirk et al. 2001). 

For many species, antifungal susceptibility data has also been provided. This has 

been derived from both the literature and in-house data from Australian clinical isolates 

generated by using the CLSI M27-A2 protocol for yeasts and the CLSI M38-A protocol 

for moulds. This composite data is provided as a guide only. MIC 90 s for Aspergillus, 

Candida, Cryptococcus and Scedosporium species are provided from large Australian 

studies based predominantly on primary isolates. In many cases the clinical relevance 

of in vitro antifungal susceptibility results remains difficult to interpret, and expert advice 

from a consulting microbiologist or infectious disease specialist may be required. 

Risk group (RG) recommendations are based on published data and on current laboratory 

safety procedures in accordance with the Australian/New Zealand Standard AS/ 

NZS 2243.3:2002. Safety in laboratories Part 3: Microbiological aspects and containment 

facilities. 

David Ellis BSc (Hons), MSc, PhD, FASM, FRCPA (Hon). 

Associate Professor 

School of Molecular & Biomedical Sciences 

University of Adelaide 

Head, Mycology Unit 

Women’s and Children’s Hospital 

North Adelaide, Australia 5006 

September, 2007 

PREFACE

Absidia corymbifera 1 

Acremonium 2 

Acrophialophora fusispora 3 

Alternaria 4 

Aphanoascus fulvescens 5 

Apophysomyces elegans 6 

Aspergillus 8 

Aspergillus flavus 9 

Aspergillus fumigatus 10 

Aspergillus nidulans 11 

Aspergillus niger 12 

Aspergillus terreus 13 

Aureobasidium pullulans 14 

Basidiobolus ranarum 15 

Beauveria 16 

Bipolaris 17 

Blastomyces dermatitidis 19 

Candida 20 

Candida albicans 23 

Candida colliculosa 24 

Candida dubliniensis 25 

Candida fabianii 26 

Candida famata 27 

Candida glabrata 28 

Candida guilliermondii 29 

Candida haemulonii 30 

Candida inconspicua 31 

Candida kefyr 32 

Candida krusei 33 

Candida lipolytica 34 

Candida lusitaniae 35 

Candida norvegensis 36 

Candida parapsilosis 37 

Candida pelliculosa 38 

Candida rugosa 39 

Candida tropicalis 40 

Chaetomium 41 

Chrysosporium tropicum 42 

Cladophialophora bantiana 43 

Cladophialophora carrionii 44 

Cladosporium 45 

Coccidioides immitis 46 

Colletotrichum 48 

Conidiobolus coronatus 49 

Cryptococcus 50 

Cryptococcus albidus 51 

Cryptococcus laurentii 51 

Cryptococcus gattii 52 

Cryptococcus neoformans 53 

CONTENTS 

Cunninghamella bertholletiae 55 

Curvularia 57 

Cylindrocarpon 58 

Drechslera 59 

Epicoccum purpurascens 60 

Epidermophyton floccosum 61 

Exophiala dermatitidis 62 

Exophiala jeanselmei complex 63 

Exophiala spinifera complex 65 

Exserohilum 66 

Fonsecaea 67 

Fusarium 68 

Fusarium oxysporum 69 

Fusarium solani 70 

Geotrichum candidum 71 

Geotrichum capitatum 72 

Gliocladium 73 

Graphium 74 

Histoplasma capsulatum 75 

Hortaea werneckii 77 

Lasiodiplodia theobromae 78 

Lecythophora hoffmannii 79 

Madurella grisea 80 

Madurella mycetomatis 81 

Malassezia 82 

Malbranchea 83 

Microsporum 84 

Microsporum audouinii 85 

Microsporum canis 86 

Microsporum canis var. distortum 88 

Microsporum canis var. equinum 89 

Microsporum cookei 90 

Microsporum ferrugineum 91 

Microsporum fulvum 92 

Microsporum gallinae 93 

Microsporum gypseum 94 

Microsporum nanum 95 

Microsporum persicolor 96 

Mortierella wolfii 97 

Mucor 98 

Mucor amphibiorum 99 

Mucor circinelloides 100 

Mucor indicus 100 

Mucor ramosissimus 100 

Nattrassia mangiferae 101 

Ochroconis gallopava 102 

Onychocola canadensis 103 

Paecilomyces 104 

Paecilomyces lilacinus 105

Paecilomyces variotii. 106 

Paracoccidioides brasiliensis 107 

Penicillium 108 

Penicillium marneffei 110 

Phaeoacremonium parasiticum 111 

Phialophora 112 

Phialophora richardsiae 112 

Phialophora verrucosa 113 

Phoma 114 

Pithomyces 115 

Prototheca 116 

Ramichloridium 117 

Ramichloridium schulzeri 117 

Rhinocladiella 118 

Rhinocladiella atrovirens 118 

Rhizomucor 119 

Rhizomucor miehei 119 

Rhizomucor pusillus 120 

Rhizopus 121 

Rhizopus azygosporus 122 

R. microsporus var. microsporus 122 

R. microsporus var. oligosporus 123 

R. microsporus var. rhizopodiformis 123 

Rhizopus oryzae 124 

Rhodotorula 125 

Rhodotorula glutinis 126 

Rhodotorula mucilaginosa 127 

Saccharomyces cerevisiae 128 

Saksenaea vasiformis 129 

Scedosporium apiospermum 131 

Scedosporium aurantiacum 131 

Scedosporium prolificans 134 

Schizophyllum commune 135 

Scopulariopsis 136 

Sepedonium 137 

Sporothrix schenckii 138 

Stemphylium 140 

Syncephalastrum 141 

Trichoderma 142 

Trichophyton 143 

Trichophyton ajelloi 144 

Trichophyton concentricum 145 

Trichophyton equinum 147 

Trichophyton erinacei 149 

Trichophyton interdigitale 151 

T. interdigitale var. nodulare 153 

Trichophyton mentagrophytes 154 

T. mentag. var. quinckeanum 156 

Trichophyton rubrum 158 

CONTENTS 

Trichophyton rubrum granular type 160 

Trichophyton schoenleinii 162 

Trichophyton soudanense 163 

Trichophyton terrestre 164 

Trichophyton tonsurans 165 

Trichophyton verrucosum 167 

Trichophyton violaceum 169 

Trichosporon 170 

Trichosporon asahii 171 

Trichosporon asteroides 172 

Trichosporon cutaneum 172 

Trichosporon inkin 172 

Trichosporon mucoides 173 

Trichosporon ovoides 173 

Trichothecium roseum 174 

Ulocladium 175 

Veronaea botryosa 176 

Verticillium 177 

Microscopy Stains & Techniques 178 

Calcofluor White with 10% KOH 178 

KOH with Chlorazol Black 178 

India Ink Mounts 178 

Lactophenol Cotton Blue (LPCB) 179 

Direct Microscopic Preparations 179 

Cellotape Flag Preparations 179 

Slide Culture Preparations 180 

Specialised Culture Media 181 

Bird seed agar 181 

Bromcresol Purple Milk Agar 181 

CDBT media 182 

CGB media 182 

Cornmeal agar 183 

Cornmeal glucose sucrose agar 183 

Czapek Dox agar 183 

Dixon’s agar 183 

Hair perforation test 184 

Lactritmel agar 184 

Littman oxgall agar 184 

Malt extract agar 185 

1% Peptone agar 185 

Potato dextrose agar 185 

Rice grain slopes 185 

Sabouraud dextrose agar 186 

Sabouraud dextrose agar 5% NaCl 186 

Tap water agar 187 

Urease agar with 0.5% glucose 187 

Vitamin free agar 187 

References 188

Descriptions of Medical Fungi 

Absidia corymbifera (Cohn) Saccardo & Trotter 

The genus Absidia is characterised by a differentiation of the hyphae into arched stolons 

bearing more or less verticillate sporangiophores at the raised part of the stolon 

(internode), and rhizoids formed at the point of contact with the substrate (at the node). 

This feature separates species of Absidia from the genus Rhizopus, where the sporangia 

arise from the nodes and are therefore found opposite the rhizoids. The sporangia 

are relatively small, globose, pyriform or pear-shaped and are supported by a 

characteristic funnel-shaped apophysis. This distinguishes Absidia from the genera 

Mucor and Rhizomucor, which have large, globose sporangia without an apophysis. 

Absidia currently contains 21 mostly soil-borne species. A. corymbifera is a known 

human pathogen causing pulmonary, rhinocerebral, disseminated, CNS or cutaneous 

zygomycosis. 

Colonies are fast growing, floccose, white at first becoming pale grey with age, and 

up to 1.5 cm high. Sporangiophores are hyaline to faintly pigmented, simple or sometimes 

branched, arising solitary from the stolons, in groups of three, or in whorls of up 

to seven. Rhizoids are very sparingly produced and may be difficult to find without the 

aid of a dissecting microscope to examine the colony on the agar surface. Sporangia 

are small (10-40 µm in diameter) and are typically pyriform in shape with a characteristic 

conical-shaped columella and pronounced apophysis, often with a short projection 

at the top. Sporangiospores vary from subglobose to oblong-ellipsoidal (3-7 x 2.5-4.5 

µm), hyaline to light grey and smooth-walled. Temperature: optimum 35-37 O C; maximum 

45 O C. RG-2 organism. 

For descriptions of species, keys to taxa and additional information see Ellis and Hesseltine 

(1965 and 1966), Hesseltine and Ellis (1964 and 1966), Nottebrock et al. (1974), 

O’Donnell (1979), Samson et al. (1995), Domsch et al. (1980), McGinnis (1980), de 

Hoog et al. (2000) and Ellis (2005b). 

Key Features: zygomycete, small pyriform-shaped 

sporangia with a characteristic conical-shaped columellae 

and pronounced apophysis, rapid growth at 

40 O C. 

Antifungal 

MIC µg/mL 

Range MIC90 Amphotericin B 0.03-2 1 

Flucytosine >256 >256 

Fluconazole >16 >16 

Itraconazole 0.03-2 0.5 

Posaconazole 0.03 - 1 0.25 

Voriconazole 2->64 >16 

Very limited data, antifungal susceptibility testing 

of individual strains is recommended. Sun et 

al. (2002), Dannaoui et al. (2003), Espinel-Ingroff 

et al. (2001), Espinel-Ingroff (2003, 2006), Singh 

et al. (2005), Sabatelli et al. (2006) and WCH inhouse 

data. 

15 µm 

1 

A. corymbifera showing a typical 

pyriform-shaped sporangium 

with a conical-shaped columella 

and pronounced apophysis (arrow).

2 


Acremonium Link ex Fries 

Colonies are usually slow growing, often compact and moist at first, becoming powdery, 

suede-like or floccose with age, and may be white, grey, pink, rose or orange in 

colour. Hyphae are fine and hyaline and produce mostly simple awl-shaped erect phialides. 

Conidia are usually one-celled (ameroconidia), hyaline or pigmented, globose 

to cylindrical, and mostly aggregated in slimy heads at the apex of each phialide. 

The genus Acremonium currently contains 100 species, most are saprophytic being 

isolated from dead plant material and soil. However a number of species including A. 

falciforme, A. kiliense, A. recifei, A. alabamensis, A. roseogriseum and A. strictum are 

recognised as opportunistic pathogens of man and animals, causing mycetoma, mycotic 

keratitis and onychomycosis. RG-2 for species isolated from humans. 

Microconidial Fusarium isolates may be confused with Acremonium, but they usually 

grow faster and have colonies with a characteristic fluffy appearance. 

Key Features: hyphomycete with solitary, erect, hyaline, awl-shaped phialides producing 

single-celled, globose to cylindrical conidia, mostly in slimy heads. 

For descriptions of species, keys to taxa and additional information see Gams (1971), 

Domsch et al. (1980), Samson et al. (1995) and de Hoog et al. (2000). 

10 µm 

Acremonium showing long awl-shaped phialides producing cylindrical, 

one-celled conidia mostly aggregated in slimy heads at the 

apex of each phialide. 

Antifungal 

MIC µg/mL 

Range 

Antifungal 

MIC µg/mL 

Range 

Itraconazole 0.5->8 Amphotericin B 0.5-16 

Posaconazole 0.06-4 Caspofungin 0.03->8 

Voriconazole 0.06-4 Anidulafungin 0.5->8 

Very limited data, antifungal susceptibility testing of individual strains 

is recommended. Guarro et al. (1997), Pfaller et al. (1998, 2002a), 

Espinel-Ingroff (2003), Cuenca-Estrella et al. (2006) and WCH inhouse 

data.

Descriptions of Medical Fungi 3 

Acrophialophora fusispora (S.B. Saksena) Samson 

Colonies fast growing, greyish-brown with a black reverse. Conidiophores arising singly, 

terminally and laterally from the hyphae, erect, straight or slightly flexuose, tapering 

towards the apex, pale brown, rough-walled, up to 15 μm long, 2-5 μm wide, with 

whorls of phialides on the upper part. Phialides flask-shaped with a swollen base 

and a long, narrow neck, hyaline, smooth-walled or echinulate, 9-15 x 3-4.5 μm in the 

broadest part. Conidia in long chains, limoniform, one-celled, pale brown 5-12 x 3-6 

μm, smooth to finely echinulate with indistinct spiral bands. Temperature: optimum 

40 O C; maximum 50 O C. 

The genus Acrophialophora contains 3 species and is most commonly associated with 

soil, especially from India. A. fusispora is a rare human pathogen. RG-1 organism. 

Key Features: hyphomycete with flask-shaped phialides producing long chains of 

one-celled, limoniform, pale brown conidia, with indistinct spiral bands. 

For descriptions of species, keys to taxa and additional information see Domsch et al. 

(1980), de Hoog et al. (2000) and Al-Mohsen et al. (2000). 

10 μm 

Culture, phialides and conidia with striations (arrows) of A. fusispora. 

Antifungal 

MIC µg/mL 

Range 

Antifungal 

MIC µg/mL 

Range 

Fluconazole 8-32 Amphotericin B 0.25-2 

Itraconazole 0.06-0.125 Flucytosine >64 

Voriconazole 0.06 Posaconazole 0.03 

Very limited data, antifungal susceptibility testing of individual strains 

is recommended. Al-Mohsen et al. (2000) and WCH in-house data.

4 


Colonies are fast growing, black to olivaceous-black or greyish, and are suede-like to 

floccose. Microscopically, branched acropetal chains (blastocatenate) of multicellular 

conidia (dictyoconidia) are produced sympodially from simple, sometimes branched, 

short or elongate conidiophores. Conidia are obclavate, obpyriform, sometimes ovoid 

or ellipsoidal, often with a short conical or cylindrical beak, pale brown, smooth-walled 

or verrucose. 

The genus contains 44 species, most are plant parasites, but a few species are ubiquitous 

and are also frequently soil-borne. A. alternata is the most common of these. 

Although usually seen as saprophytic contaminants, Alternaria species are recognised 

causative agents of mycotic keratitis. 

Alternaria species soon lose their ability to sporulate in culture. Potato dextrose agar 

and cornmeal agar are the most suitable media to use, and incubation under near 

ultra-violet light is recommended to maintain sporulation. Temperature: optimum 25- 

28 O C; maximum 31-32 O C. RG-1 organism. 

Key Features: dematiaceous hyphomycete producing chains of darkly pigmented, 

ovoid to obclavate dictyoconidia, often with short conical or cylindrical beaks. 

For descriptions of species, keys to taxa and additional information see Simmons (1967), 

Ellis (1971), Domsch et al. (1980), Samson et al. (1995), de Hoog et al. (2000). 

Antifungal 

Alternaria Nees ex Fries 

20 μm 

Alternaria alternata showing branched acropetal chains and multi-celled, 

obclavate to obpyriform conidia with short conical beaks. 

MIC µg/mL 

MIC µg/mL 

Range MIC90 Antifungal 

Range MIC90 Fluconazole 16->64 >64 Amphotericin B 0.125->16 2 (4) 

Itraconazole 0.125-2 1 Flucytosine >128 >128 

Voriconazole 0.5-2 1 Posaconazole 0.06-0.25 0.25 

Limited data, antifungal susceptibility testing of individual strains is recommended. 

McGinnis and Pasarell (1998), Pujol et al. (2000), Espinel-Ingroff et al. (2001), 

Espinel-Ingroff (2003), Sabatelli et al. (2006) and WCH in-house data.


Aphanoascus fulvescens (Cooke) Apinis 

Colonies are moderately fast growing, white to tan with the production of numerous 

spherical, pseudoparenchymatous, buff to light brown cleistothecia (non-ostiolate 

ascocarps). Asci are subspherical to ellipsoidal and eight-spored. Ascospores light 

brown, yellowish to pale brown in mass, irregularly reticulate, lens-shaped, 3.5-4.7 x 

2.5-3.5 µm. Aphanoascus fulvescens has a Chrysosporium anamorph showing typical 

pyriform to clavate-shaped conidia with truncated bases, 15.0-17.5 x 3.7-6.0 µm, 

which are formed either intercalary, laterally or terminally. 

Aphanoascus fulvescens is a soil keratinolytic ascomycete which occasionally causes 

dermatomycosis in man and animals. RG-2 organism. 

Key Features: keratinolytic ascomycete with a Chrysosporium anamorph. 


(1980), McGinnis (1980) and de Hoog et al. (2000). 

100 μm 

10 μm 

Culture, cleistothecium and conidia of Aphanoascus fulvescens.

6 


Apophysomyces elegans Misra, Srivastava & Lata 

Colonies are fast growing, white, becoming creamy white to buff with age, downy with 

no reverse pigment, and are composed of broad, sparsely septate (coenocytic) hyphae 

typical of a zygomycetous fungus. Sporangiophores are unbranched, straight or 

curved, slightly tapering towards the apex, up to 200 µm long, 3- 5 µm in width near 

the apophysis, and hyaline when young but developing a light to dark brown pigmentation 

and a conspicuous sub-apical thickening 10-16 µm below the apophysis with age. 

Sporangiophores arise at right angles from the aerial hyphae and often have a septate 

basal segment resembling the “foot cell” commonly seen in Aspergillus. Rhizoids are 

thin-walled, subhyaline and predominantly unbranched. Sporangia are multispored, 

small (20-50 µm diameter), typically pyriform in shape, hyaline at first, sepia-coloured 

when mature, columellate and strongly apophysate. Columellae are hemispherical in 

shape and the apophyses are distinctively funnel or bell-shaped. Sporangiospores are 

smooth-walled, mostly oblong, occasionally subglobose, (3-4 x 5-6 µm), subhyaline to 

sepia in mass. Good growth at 26 O C, 37 O C and 42 O C. RG-2 organism. 

Apophysomyces elegans is readily distinguishable from other zygomycetes of medical 

importance, especially the morphologically similar, strongly apophysate pathogen 

Absidia corymbifera, by having sporangiophores with distinctive funnel or bell-shaped 

apophyses and hemispherical-shaped columellae. In addition, there is a conspicuous 

pigmented sub-apical thickening which constricts the lumen of the sporangiophore 

below the apophysis, and there are also distinctive foot cells. 

Laboratory identification of this fungus may be difficult or delayed because of the 

mould’s failure to sporulate on the primary isolation media or on subsequent subculture 

onto potato dextrose agar. Sporulation may be stimulated by the use of nutrient 

deficient media, like cornmeal-glucose-sucrose-yeast extract agar, Czapek Dox agar, 

or by using the agar block method described by Ellis and Ajello (1982) and Ellis and 

Kaminski (1985). 

Key Features: zygomycete, rare human pathogen usually associated with invasive 

lesions following the traumatic implantation of the fungus through the skin. Soil fungus 

with a tropical to sub-tropical distribution. Characteristic “cocktail glass” apophysate 

sporangial morphology with conspicuous sub-apical thickening of the sporangiophore, 

rapid growth at 42 O C. 

For descriptions of species, keys to taxa and additional information see Cooter et al. 

(1990), Ellis and Ajello, (1982), Misra et al. (1979), Padhye and Ajello (1988), Lawrence 

et al. (1986), Wieden et al. (1985), de Hoog et al. (2000) and Ellis (2005b). 

Antifungal 

MIC µg/mL 

MIC µg/mL 

Antifungal 

Range Range 

Fluconazole >64 Amphotericin B 0.03-2 

Itraconazole 0.03-8 Flucytosine >256 

Posaconazole 0.03-4 Voriconazole 8->64 

Very limited data, antifungal susceptibility testing of individual strains is 

recommended. Sun et al. (2002), Dannaoui et al. (2003), Sabatelli et al. 

(2006) and WCH in-house data.


Apophysomyces elegans Misra, Srivastava & Lata 

10 μm 10 μm 

a b 

(a) Young, multispored, pyriform sporangium of A. elegans showing a typical funnel-shaped 

apophysis but without the sub-apical thickening of a more mature 

sporangiophore. (b) Mature sporangium of A. elegans showing distinctive funnel-shaped 

apophyses, columellae, and a conspicuous pigmented sub-apical 

thickening which constricts the lumen of the sporangiophore below the apophysis 

(arrow). Sporangiospores are smooth-walled, oblong and subhyaline.

8 


Aspergillus Micheli ex Link 

Colonies are usually fast growing, white, yellow, yellow-brown, brown to black or 

shades of green, mostly consisting of a dense felt of erect conidiophores. Conidiophores 

terminate in a vesicle covered with either a single palisade-like layer of phialides 

(uniseriate) or a layer of subtending cells (metulae) which bear small whorls of 

phialides (the so called biseriate structure). The vesicle, phialides, metulae (if present) 

and conidia form the conidial head. Conidia are one-celled, smooth or rough walled, 

hyaline or pigmented are produced in long dry chains which may be divergent (radiate) 

or aggregated in compact columns (columnar). Some species may produce Hülle cells 

or sclerotia. 

For identification, isolates are usually inoculated at three points on Czapek Dox agar 

and 2% malt extract agar and incubated at 25 O C. Most species sporulate within 7 

days. Descriptions are primarily based on colony pigmentation and morphology of the 

conidial head. Microscopic mounts are best made using cellotape flag or slide culture 

preparations mounted in lactophenol cotton blue. A drop of alcohol is usually needed 

to remove bubbles and excess conidia. 

Key Features: hyaline hyphomycete showing distinctive conidial heads with flaskshaped 

phialides arranged in whorls on a vesicle. 

For descriptions of species, keys to taxa and additional information see Raper and 

Fennell (1965), Domsch et al. (1980), McGinnis (1980), Onions et al. (1981), Samson 

and Pitt (1990), Samson et al. (1995), de Hoog et al. (2000) and Klich (2002). 

a 

conidia 

phialides 

vesicle 

stipe 

metulae 

Conidial head morphology in Aspergillus (a) uniseriate, (b) biseriate. 

b


On Czapek Dox agar, colonies are granular, 

flat, often with radial grooves, yellow 

at first but quickly becoming bright to dark 

yellow-green with age. Conidial heads 

are typically radiate, later splitting to form 

loose columns (mostly 300-400 µm in diameter), 

biseriate but having some heads 

with phialides borne directly on the vesicle 

(uniseriate). Conidiophore stipes are 

hyaline and coarsely roughened, often 

more noticeable near the vesicle. Conidia 

are globose to subglobose (3-6 µm in 

diameter), pale green and conspicuously 

echinulate. Some strains produce brownish 

sclerotia. 

A. flavus has a world-wide distribution and 

normally occurs as a saprophyte in soil 

and on many kinds of decaying organic 

matter, however, it is also a recognised 

pathogen of humans and animals. RG-2 

organism. 

Key Features: spreading yellow-green 

colonies, rough-walled stipes, mature 

vesicles bearing phialides over their entire 

surface and conspicuously echinulate 

conidia. 

Antifungal 

MIC µg/mL 

Range MIC90 Amphotericin B 0.06->8 4 


Voriconazole 0.03-2 0.5 

Posaconazole 0.03-1 0.5 

Anidulafungin

10 


On Czapek Dox agar, colonies show typical 

blue-green surface pigmentation with 

a suede-like surface consisting of a dense 

felt of conidiophores. Conidial heads are 

typically columnar (up to 400 x 50 µm 

but often much shorter and smaller) and 

uniseriate. Conidiophore stipes are short, 

smooth-walled and have conical-shaped 

terminal vesicles which support a single 

row of phialides on the upper two thirds 

of the vesicle. Conidia are produced in 

basipetal succession forming long chains 

and are globose to subglobose (2.5-3.0 

µm in diameter), green and rough-walled 

to echinulate. Note: This species is thermotolerant 

and grows at temperatures up 

to 55 O C. 

A. fumigatus is truly a cosmopolitan mould 

and has been found almost everywhere 

on every conceivable type of substrate. It 

is an important pathogen of humans and 

animals. RG-2 organism. 

Key Features: uniseriate and columnar 

conidial heads with the phialides limited 

to the upper two thirds of the vesicle and 

curving to be roughly parallel to each 

other. 

Antifungal 

MIC µg/mL 

Range MIC90 Amphotericin B 0.03->8 2 

Itraconazole 16 0.5 

Voriconazole


Aspergillus nidulans (Eidam) Wint. 

Teleomorph: Emericella nidulans (Eidam) Vuill. 

On Czapek Dox agar, colonies are typically plain green in colour with dark red-brown 

cleistothecia developing within and upon the conidial layer. Reverse may be olive to 

drab-grey or purple-brown. Conidial heads are short columnar (up to 70 x 30 µm in 

diameter) and biseriate. Conidiophore stipes are usually short, brownish and smoothwalled. 

Conidia are globose (3-3.5 µm in diameter) and rough-walled. 

A. nidulans is a typical soil fungus with a world-wide distribution, it has also been reported 

causing disease in human and animals. RG-1 organism. 

Key Features: conidial heads are short columnar and biseriate. Stipes are usually 

short, brownish and smooth-walled. Conidia are globose and rough-walled. 

20 μm 

10 μm 20 μm 

a b c 

(a) Cleistothecium of Emericella nidulans (anamorph Aspergillus nidulans) showing 

numerous reddish-brown ascospores and thick-walled hülle cells; (b) cleistothecia are 

often surrounded by a mass of hülle cells which are up to 25 µm in diameter; (c) conidial 

head and stipe and (d) culture of A. nidulans. 

Antifungal 

MIC µg/mL 





Caspofungin 0.125-8 nd 

Espinel-Ingroff et al. (2001), Espinel-Ingroff 

(2003), Cuenca-Estrella et al. (2006). 

MIC s from Australian clinical isolates (nd 

90 

= not done). 

d

12 


On Czapek Dox agar, colonies consist of 

a compact white or yellow basal felt covered 

by a dense layer of dark-brown to 

black conidial heads. Conidial heads are 

large (up to 3 mm by 15 to 20 µm in diameter), 

globose, dark brown, becoming 

radiate and tending to split into several 

loose columns with age. Conidiophore 

stipes are smooth-walled, hyaline or turning 

dark towards the vesicle. Conidial 

heads are biseriate with the phialides 

borne on brown, often septate metulae. 

Conidia are globose to subglobose (3.5-5 

µm in diameter), dark brown to black and 

rough-walled. 

A. niger is one of the most common and 

easily identifiable species of the genus 

Aspergillus, with its white to yellow mat 

later bearing black conidia. This species 

is very commonly found in aspergillomas 

and is the most frequently encountered 

agent of otomycosis. It is also a common 

laboratory contaminant. RG-1 organism. 

Key Features: conidial heads are dark 

brown to black, radiate and biseriate with 

metulae twice as long as the phialides. 

Conidia brown and rough-walled. 

Antifungal 

MIC µg/mL 


Itraconazole 0.03->8 0.5 

Voriconazole


On Czapek Dox agar, colonies are typically 

suede-like and cinnamon-buff to 

sand brown in colour with a yellow to deep 

dirty brown reverse. Conidial heads are 

compact, columnar (up to 500 x 30-50 µm 

in diameter) and biseriate. Conidiophore 

stipes are hyaline and smooth-walled. 

Conidia are globose to ellipsoidal (1.5-2.5 

µm in diameter), hyaline to slightly yellow 

and smooth-walled. 

A. terreus occurs commonly in soil and is 

occasionally reported as a pathogen of 

humans and animals. RG-2 organism. 

Key Features: cinnamon-brown cultures, 

conidial heads biseriate with metulae as 

long as the phialides. 

For descriptions of species, keys to taxa 

and additional information see Raper and 

Fennell (1965), Domsch et al. (1980), 

McGinnis (1980), Onions et al. (1981), 

Samson and Pitt (1990), Samson et al. 

(1995), de Hoog et al. (2000) and Klich 

(2002). 

Antifungal 

MIC µg/mL 





Anidulafungin 0.03 nd 

Caspofungin 0.015-0.5 nd 

Espinel-Ingroff et al. (2001), Pfaller et 

al. (2002), Diekema et al. (2003), Espinel-Ingroff 

(2003), Serrano et al. (2003), 

Cuenca-Estrella et al. (2006). MIC s 90 

from Australian clinical isolates (nd = not 

done). 

Aspergillus terreus Thom 

10 μm 

Culture and conidial head and conidiophore 

of A. terreus. Note: conidial heads 

are biseriate.

14 


Aureobasidium pullulans (de Bary) Arnaud 

Colonies are fast growing, smooth, soon covered with slimy masses of conidia, cream 

or pink to brown or black. Hyphae hyaline and septate, frequently becoming darkbrown 

with age and forming chains of one- to two-celled, thick-walled, darkly pigmented 

arthroconidia. These arthroconidia actually represent the Scytalidium anamorph of 

Aureobasidium and are only of secondary importance in recognising members of this 

genus. Conidia are produced synchronously in dense groups from indistinct scars or 

from short denticles on undifferentiated, hyaline to sub-hyaline hyphae. Conidia are 

hyaline, smooth-walled, single-celled, ellipsoidal but of very variable shape and size 

(8-12 x 4-6 µm), often with an indistinct hilum (= a mark or scar at the point of attachment). 

Temperature: optimum 25 O C; maximum 35-37 O C. 

This species has two varieties: A. pullulans var. pullulans, with a colony which remains 

pink, light brown, or yellow for at least three weeks, and A. pullulans var. melanogenum 

which soon becomes black or greenish-black due to dark hyphae which often fall apart 

into separate cells. A. pullulans has a world-wide distribution and is usually isolated 

as a saprophyte, occasionally from skin and nails. However, it has also been reported 

as a rare causative agent of phaeohyphomycosis, mycotic keratitis and peritonitis in 

patients on continuous ambulatory peritoneal dialysis (CAPD). RG-1 organism. 

Key Features: hyphomycete (so called black yeast) producing hyaline blastoconidia 

simultaneously from the vegetative hyphae, which may also form chains of darkly pigmented, 

thick-walled arthroconidia. 

For descriptions of species, keys to taxa and additional information see Hermanides- 

Nijhof (1977), Domsch et al. (1980), McGinnis (1980) and de Hoog et al. 2000. 

20 μm 

A. pullulans showing chains of one- to two-celled, darkly pigmented arthroconidia of 

the Scytalidium anamorph of Aureobasidium and the presence of numerous hyaline, 

single-celled, ovoid-shaped conidia which are produced on short denticles. 

Antifungal 

MIC µg/mL MIC µg/mL MIC µg/mL 

Antifungal Antifungal 

Range Range Range 

Amphotericin B 0.125-2 Itraconazole 0.03-0.25 Voriconazole 0.03-0.5 

Very limited data, antifungal susceptibility testing of individual strains is recommended. 

McGinnis and Pasarell (1998), Espinel-Ingroff et al. (2001) and WCH in-house data.


Synonyms: Basidiobolus meristosporus Drechsler; Basidiobolus heterosporus Srinivasan 

& Thirumalachar; Basidiobolus haptosporus Drechsler. 

Colonies are moderately fast growing at 30 O C, flat, yellowish-grey to creamy-grey, glabrous, 

becoming radially folded and covered by a fine, powdery, white surface mycelium. 

Note: satellite colonies are often formed by germinating conidia ejected from 

the primary colony. Microscopic examination usually shows the presence of large 

vegetative hyphae (8-20 µm in diameter) forming numerous round (20-50 µm in diameter), 

smooth, thick-walled zygospores that have two closely appressed beak-like 

appendages. The production of “beaked” zygospores is diagnostic for the genus. Two 

types of asexual conidia are formed, although isolates often lose their sporulating ability 

with subculture and special media incorporating glucosamine hydrochloride and 

casein hydrolsate may be needed to stimulate sporulation (Shipton and Zahari, 1987). 

Primary conidia are globose, one-celled, solitary and are forcibly discharged from a 

sporophore. The sporophore has a distinct swollen area just below the conidium that 

actively participates in the discharge of the conidium. Secondary (replicative) conidia 

are clavate, one-celled and are passively released from a sporophore. These sporophores 

are not swollen at their bases. The apex of the passively released spore has 

a knob-like adhesive tip. These spores may function as sporangia, producing several 

sporangiospores. RG-2 organism. 

Basidiobolus ranarum is commonly present in decaying fruit and vegetable matter, and 

as a commensal in the intestinal tract of frogs, toads and lizards. It has been reported 

from tropical Africa, India, Indonesia and South East Asia including Australia. 

For descriptions of species, keys to taxa and additional information see Strinivasan and 

Thirumalachar (1965), Greer and Friedman (1966), Dworzack et al. (1978), McGinnis 

(1980), King (1983), Rippon (1988), Davis et al. (1994), Jong and Dugan (2003), de 

Hoog et al. (2000) and Ellis (2005a). 

20 μm 

Basidiobolus ranarum Eidem 

a 20 μm 

b 

(a) Sporophore and conidia and (b) zygospores of Basidiobolus ranarum.

16 


Colonies are usually slow growing, mostly not exceeding 2 cm in ten days at 20 O C, 

downy, at first white but later often becoming yellow to pinkish. The genus Beauveria 

is characterised by the sympodial development of single-celled conidia (ameroconidia) 

on a geniculate or zig-zag rachis. Conidiogenous cells are flask-shaped, rachiform, 

proliferating sympodially and are often aggregated into sporodochia or synnemata. 

Conidia are hyaline and globose or ovoid in shape. RG-1 organism. 

Three species are recognised, two of which are well known parasites of insects. B. 

bassiana is the most common species and is best known as the causal agent of the 

disastrous muscardine in silkworms. Beauveria species are occasionally isolated in 

the clinical laboratory as saprophytic contaminants. 

Key Features: hyphomycete showing sympodial development of ameroconidia on a 

geniculate or zig-zag rachis emanating from a flask-shaped conidiophore. 

For descriptions of species, keys to taxa and additional information see de Hoog (1972), 

Domsch et al. (1980), McGinnis (1980) and de Hoog et al. (2000). 

20 μm 

Beauveria Vuillemin 

Beauveria bassiana showing sympodial development of conidia on a geniculate 

or zig-zag rachis. Conidiogenous cells are flask-shaped, rachiform, 

proliferating sympodially and are often aggregated into sporodochia or synnemata. 

Conidia are hyaline and globose or ovoid in shape, 2-3 mm diameter 

(phase contrast image).


Teleomorph: Cochliobolus Drechsler 

Bipolaris Shoemaker 

Colonies are moderately fast growing, effuse, grey to blackish brown, suede-like to 

floccose with a black reverse. Microscopic morphology shows sympodial development 

of pale brown pigmented, pseudoseptate conidia on a geniculate or zig-zag rachis. 

Conidia are produced through pores in the conidiophore wall (poroconidia) and are 

straight, fusiform to ellipsoidal, rounded at both ends, smooth to finely roughened, germinating 

only from the ends (bipolar). 

The genus Bipolaris contains about 45 species which are mostly subtropical and tropical 

plant parasites; however several species, notably B. australiensis, B. hawaiiensis 

and B. spicifera are well documented human pathogens. RG-1 organisms. 

Key Features: dematiaceous hyphomycete producing sympodial, pseudoseptate, pale 

brown, straight, fusiform to ellipsoidal poroconidia, which are rounded at both ends. 

The genera Drechslera, Bipolaris, Curvularia and Exserohilum are all closely related 

and differentiation of the genera relies upon a combination of characters including conidial 

shape, the presence or absence of a protruding hilum, the contour of the basal 

portion of the conidium and its hilum, the point at which the germ tube originates from 

the basal cell and, to a lesser degree, the sequence and location of the first three conidial 

septa. The table below is modified from Domsch et al. (1980). 

Anamorph Main characters Teleomorph 

Drechslera Conidia cylindrical, germinating from any cell, 

hilum not protuberant 

Bipolaris Conidia fusiform-ellipsoidal, central cells not 

much darker and broader than the distal ones, 

hilum not protuberant, germination bipolar. 

Curvularia Conidia with 2-3 broader and darker central 

cells, often curved, with or without a prominent 

hilum, germination bipolar. 

Exserohilum Conidia fusiform-cylindrical to obclavate, with 

a protuberant hilum germination bipolar. 

Pyrenophora 

Cochliobolus 

Cochliobolus 

Setosphaeria 

Species of Bipolaris, Curvularia and Exserohilum are causative agents of phaeohyphomycosis 

which is an emerging mycotic infection of humans and lower animals 

caused by a number of dematiaceous (brown-pigmented) fungi where the tissue morphology 

of the causative organism is mycelial. This separates it from other clinical 

types of disease involving brown-pigmented fungi where the tissue morphology of the 

organism is a grain (mycotic mycetoma) or sclerotic body (chromoblastomycosis). 

For descriptions of species, keys to taxa and additional information see Ellis (1971 

and 1976), Luttrell (1978), Domsch et al. (1980), Alcorn (1983), Padhye et al. (1986), 

McGinnis et al. (1986b), Sivanesan (1987), Rippon (1988) and de Hoog et al. (2000). 

Also see Descriptions for Curvularia, Drechslera and Exserohilum.

18 


10 μm 

Bipolaris australiensis showing sympodial development of pale 

brown, fusiform to ellipsoidal, pseudoseptate, poroconidia on a 

geniculate or zig-zag rachis. 

Antifungal 

Bipolaris Shoemaker 

MIC µg/mL 

MIC µg/mL 

Range 

Antifungal 

Range 

Itraconazole 0.03-1 Amphotericin B 0.06-2 

Posaconazole 0.06-0.05 Anidulafungin 1-4 

Voriconazole 0.06-0.05 Caspofungin 1-4 


Espinel-Ingroff et al. (2001), Pfaller et al. (2002a), Espinel-Ingroff 

(2003), McGinnis and Pasarell (1998) and WCH in-house data.


Blastomyces dermatitidis Gilchrist & Stokes 

Colonies (SDA) at 25 O C are variable in morphology and rate of growth. They may grow 

rapidly, producing a fluffy white mycelium or slowly as glabrous, tan, nonsporulating 

colonies (Fig. a). Growth and sporulation may be enhanced by yeast extract. Most 

strains become pleomorphic with age. Microscopically, hyaline, ovoid to pyriform, onecelled, 

smooth-walled conidia (2-10 µm in diameter) of the Chrysosporium type, are 

borne on short lateral or terminal hyphal branches. 

Colonies on blood agar at 37 O C are wrinkled and folded, glabrous and yeast-like. Microscopically, 

the organism produces the characteristic yeast phase as seen in tissue 

pathology; ie. B. dermatitidis is a dimorphic fungus. 

WARNING: RG-3 organism. Cultures of Blastomyces dermatitidis may represent a 

biohazard to laboratory personnel and should be handled in an appropriate pathogen 

handling cabinet. In the past, conversion from the mould form to the yeast form was 

necessary to positively identify this dimorphic pathogen from species of Chrysosporium 

or Sepedonium; however, culture identification by exoantigen test is now the method 

of choice. 

Histopathology: Tissue sections show large, broad-based, unipolar budding yeastlike 

cells, which may vary in size from 8-15 µm (Fig. b), with some larger forms up to 

30 µm in diameter. Note: tissue sections need to be stained by Grocott’s methenamine 

silver method to clearly see the yeast-like cells, which are often difficult to observe in 

H&E preparations. 

Key Features: clinical history, tissue pathology, culture identification by positive 

exoantigen test. 

For descriptions of species, keys to taxa and additional information see McGinnis 

(1980), Chandler et al. (1980), Kaufman and Standard (1987) and Rippon (1988). 

Antifungal 

a b 

10 μm 

MIC µg/mL 

MIC µg/mL 

Antifungal 

Range MIC Range MIC 90 90 

Fluconazole 0.125-64 4-16 Amphotericin B 0.03-1 0.5 

Itraconazole 0.03->16 0.125-2 Voriconazole 0.03-16 0.25 

Posaconazole 0.03-2 0.125 Caspofungin 0.5-8 2 

Limited data available. Sugar and Liu (1996), Espinel-Ingroff et al. (2001), 

Espinel-Ingroff (2003), Gonzales et al. (2005) and Sabatelli et al. (2006).

20 


The genus Candida is characterised by globose to elongate yeast-like cells or blastoconidia 

that reproduce by multilateral budding, polar budding if present on a narrow 

base, pseudohyphae and occasionally true hyphae may also be present. Arthroconidia, 

ballistoconidia and colony pigmentation are always absent. Fermentation or 

not: Nitrate assimilation or not: Inositol assimilation or not, however all inositol positive 

strains form pseudohyphae. In the past, the genus Torulopsis was separated from 

the genus Candida by the absence of pseudomycelium. However, in 1978 Yarrow & 

Meyer amended the description of Candida to include all species previously included 

in Torulopsis. 

Several species of Candida may be aetiological agents, most commonly C. albicans, 

followed by C. parapsilosis, C. glabrata, C. krusei and C. tropicalis. However a number 

of other species may also be isolated (see table below). All are ubiquitous and occur 

naturally on humans. 

Identification: 

Candida Berkhout 

Ensure that you start with a fresh growing pure culture; streak for single colony isolation 

if necessary. 

Chromogenic agars are now being used for primary isolation for both the detection of 

mixed flora and rapid species identification, especially from non-sterile sites. 

Germ Tube Test. A rapid screening test for Candida albicans and Candida dubliniensis. 

0.5 mL of serum, containing 0.5% glucose, is lightly inoculated with the test organism 

and incubated at 35 O C for 2-3 hours. On microscopy, the production of germ tubes by 

the cells is diagnostic for Candida albicans. 

10 μm 

Production of germ tubes by C. albicans. 

Species distribution from 944 patients 

with candidemia (Australian 

Candidemia Study 2002-2004). 

Species No % 

C. albicans 447 47.3 

C. parapsilosis 182 19.3 

C. glabrata 167 17.8 

C. krusei 46 4.9 

C. tropicalis 46 4.9 

C. dubliniensis 22 2.3 

C. guilliermondii 11 1.2 

C. lusitaniae 8 0.8 

C. kefyr 5 0.5 

C. pelliculosa 3 0.3 

C. rugosa 2 0.2 

C. colliculosa 1 0.1 

C. famata 1 0.1 

C. inconspicua 1 0.1 

C. lipolytica 1 0.1 

C. fabianii 1 0.1


Candida Berkhout 

For the full identification of germ tube negative yeasts, morphological (Dalmau 

plate culture), physiological and biochemical tests are essential. 

(a) Dalmau Plate Culture: To set up a yeast morphology plate, dip a flamed sterilised 

straight wire into a culture to make a light inoculum and then lightly scratch the wire 

into 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 lower power of a microscope 

for the presence of pseudohyphae which may take up to 4-5 days at 26 O C to develop. 

Candida albicans also produces characteristic large, round, terminal, thick-walled vesicles 

(often called chlamydospores). The key features to remember are to use a light 

inoculum and to scratch the surface of the agar with the wire when inoculating. 

(b) Physiological and biochemical tests including fermentation and assimilation studies 

should be performed based on those used at the Centraalbureau voor Schimmelcultures, 

Utrecht, The Netherlands. Reference “The Yeasts: a taxonomic study”, edited 

by Kurtzman and Fell (1998), Elsevier Science Publishers B.V. Amsterdam. Reliable 

commercially available yeast identification kits are the API 20C, ID32C, MicroScan and 

Vitek systems. For specific identification of species see appropriate text book. 

5 mm 10 μm 

a b 

(a) Dalmau plate culture showing colonies of C. albicans growing out from scratches 

on the surface of a cornmeal/tween 80 agar plate. Note: a coverslip has been placed 

onto the agar surface covering the scratches. (b) Confirmatory test for C. albicans. 

Production of large round, thick-walled vesicles (often called chlamydospores) in 

Dalmau plate cultures. 

For descriptions of species, keys to taxa and additional information see Barnett et al. 

(1983), Kurtzman and Fell (1988) and de Hoog et al. (2000).

22 


Candida albicans (Robin) Berkhout 

10 μm 

10 μm 

CHROMagar Candida plate 

showing chromogenic colour 

change for C. albicans (green), 

C. tropicalis (blue), C. parapsilosis 

(white) and C. glabrata 

(pink). 

Candida albicans on Sabouraud’s 

dextrose agar showing 

typical cream coloured, smooth 

surfaced, waxy colonies. 

Direct smear of urine from a 

patient with candidiasis of the 

kidney showing C. albicans in 

mycelial or tissue phase with 

blastoconidia budding from the 

pseudohyphae. 

Microscopic morphology of 

C. albicans showing budding 

spherical to ovoid blastoconidia.


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 on Cornmeal and Tween 80 Agar: 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 -(s) 

Fermentation Sucrose v D-Arabinose v α-M-D-glucoside v 

Glucose + Maltose + D-Ribose -(s) D-Gluconate -(s) 

Galactose v Cellobiose - L-Rhamnose - DL-Lactate + 

Sucrose -(s) Trehalose +(s) D-Glucosamine v myo-Inositol + 

Maltose + Lactose - N-A-D-glucosamine + 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, known as C. claussenii, 

and sucrose-negative variants described as C. stellatoidea have proven to be 

synonymous with C. albicans. C. 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. 

Antifungal 

MIC µg/mL 

MIC µg/mL 

Antifungal 


Fluconazole 0.03->64 2 Amphotericin B 0.03-4 0.25 

Itraconazole 0.008->8 0.125 Flucytosine 0.03->64 0.5 

Posaconazole 0.008->8 0.016 Caspofungin 0.008->4 0.125 

Voriconazole 0.008->8 0.03 Anidulafungin 0.008->8 nd 

Good data available. Espinel-Ingroff et al. (2001), Pfaller et al. (2002b, 2006, 2007), 

Espinel-Ingroff (2003), Hajjeh et al. (2004), Richter et al. (2005) and Cuenca-Estrella 

et al. (2006). MIC 90 s from the Australian Candidemia Study (nd = not done).

24 


Candida colliculosa (Hartmann) S.A. Meyer & Yarrow 

Teleomorph: Torulaspora delbrueckii (Lindner) Lindner. 


Microscopy: Spherical to ellipsoidal budding blastoconidia, 2-6 x 3-7 µm in size. Ascospores 

may be produced on 5% malt extract or cornmeal agar after 5-30 days at 

25 O C. 


Dalmau Plate Culture on Cornmeal and Tween 80 Agar: Budding yeast cells only. 

No pseudohyphae or true hyphae produced. 


Germ Tube - L-Sorbose v L-Arabinose - D-Glucitol v 

Fermentation Sucrose v D-Arabinose - α-M-D-glucoside v 

Glucose + Maltose v D-Ribose - D-Gluconate v 

Galactose v Cellobiose - L-Rhamnose - DL-Lactate v 

Sucrose v Trehalose -,s D-Glucosamine - myo-Inositol - 

Maltose v Lactose - N-A-D-glucosamine - 2-K-D-gluconate + 

Lactose - Melibiose - Glycerol v D-Glucuronate v 

Trehalose v Raffinose v Erythritol - Nitrate - 

Assimilation Melezitose v Ribitol v Urease - 

Glucose + Soluble Starch - Galactitol - 0.1% Cycloheximide - 

Galactose v D-Xylose v D-Mannitol + Growth at 37 O C v 

Key Features: asci containing 1-4 spheroidal ascospores, variable growth at 37 O C 

and a variable sugar assimilation profile. C. colliculosa is a rare cause of candidemia. 


Antifungal 

MIC µg/mL 

MIC µg/mL 

Range Antifungal 

Range 

Fluconazole 8 Amphotericin B 0.25 

Itraconazole 0.25 Flucytosine 0.03 

Posaconazole 0.25 Caspofungin 0.06 

Voriconazole 0.06 Anidulafungin nd 

Very limited data, antifungal susceptibility testing of individual stains is recommended. 

Data from the Australian Candidemia Study (nd = not done).


Candida dubliniensis Sullivan et al. 


Microscopy: Spherical to subspherical budding blastoconidia, 3-8 x 2-7 µm in size. 



with dense verticils of blastoconidia and spherical, mostly terminal chlamydospores. 


Germ Tube + L-Sorbose - L-Arabinose - D-Glucitol + 

Fermentation Sucrose + D-Arabinose - α-M-D-glucoside +,s 

Glucose + Maltose + D-Ribose - D-Gluconate -,s 

Galactose v Cellobiose - L-Rhamnose - DL-Lactate + 

Sucrose - Trehalose + D-Glucosamine -,s myo-Inositol - 


Lactose - Melibiose v Glycerol +,s D-Glucuronate - 


Assimilation Melezitose + Ribitol +,s Urease - 


Galactose + D-Xylose v D-Mannitol + Growth at 40 O C + 

Key Features: germ tube positive, similar to C. albicans, except for absence of growth 

at 45 o C; glycerol (mostly +), methyl-α-D-glucoside (-), trehalose (-), and D-xylose (-). 

Initial colonies dark green colour on CHROMagar and producing rough colonies on 

bird seed agar. C. dubliniensis is an uncommon cause of candidemia and mucosal 

infection, especially in HIV patients. RG-2 organism. 

Antifungal 

MIC µg/mL 

MIC µg/mL 

Range MIC90 Antifungal Range MIC90 Fluconazole 0.05->64 1 Amphotericin B 0.03-2 0.125 

Itraconazole 0.008->8 0.125 Flucytosine 0.03-64 0.125 

Posaconazole 0.03-1 0.125 Caspofungin 0.008-1 0.25 

Voriconazole 0.008-2 0.016 Anidulafungin

26 


Candida fabianii (Hartmann) S.A. Meyer & Yarrow 

Teleomorph: Pichia fabianii (Wickerham) Kurtzman 


Microscopy: Spheroidal to ellipsoidal budding blastoconidia, 3.0-6.5 x 2-5.5 µm in 

size. No pseudohyphae produced. Asci when present spherical, containing 1-4 spherical, 

faintly roughened ascospores. 


Dalmau Plate Culture on Cornmeal and Tween 80 Agar: Spherical to ovoid budding 

yeast cells and occasional pseudohyphae produced. 


Germ Tube - L-Sorbose - L-Arabinose - D-Glucitol + 

Fermentation Sucrose + D-Arabinose - α-M-D-glucoside + 

Glucose + Maltose + D-Ribose - D-Gluconate + 

Galactose - Cellobiose + L-Rhamnose - DL-Lactate + 

Sucrose + Trehalose + D-Glucosamine - myo-Inositol - 

Maltose +,s Lactose - N-A-D-glucosamine - 2-K-D-gluconate - 

Lactose - Melibiose - Glycerol + D-Glucuronate - 

Trehalose - Raffinose + Erythritol - Nitrate + 

Assimilation Melezitose + Ribitol - Urease - 

Glucose + Soluble Starch + Galactitol - 0.1% Cycloheximide - 

Galactose - D-Xylose + D-Mannitol + Growth at 37 O C + 

Key Features: germ tube negative yeast and sugar assimilation pattern. Molecular 

identification may be required. Candida fabianii is a rare cause of candidemia. RG-1 

organism. 

Antifungal 

MIC µg/mL 

MIC µg/mL 

Antifungal 

Range Range 

Fluconazole 8 Amphotericin B 0.125 

Itraconazole 0.5 Flucytosine 0.03 

Posaconazole 0.5 Caspofungin 0.5 

Voriconazole 0.125 Anidulafungin nd 


Data from the Australian Candidemia Study (nd = not done).


Teleomorph: Debaryomyces hansenii (Zopf) Lodder & Kreger-van Rij. 


Microscopy: Ovoid to broadly ellipsoidal budding blastoconidia, 3.5-5 x 2-3.5 µm in 

size. No pseudohyphae produced. Asci when present spherical, persistent, containing 

1-2 spherical ascospores with rough walls. 



yeast cells only. No pseudohyphae produced. 

Key Features: germ tube negative yeast and sugar assimilation pattern. Candida 

famata is a common environmental isolate, however it is only rarely recovered from 

clinical specimens, usually associated with skin. RG-1 organism. 

Antifungal 

Candida famata (Harrison) S.A. Meyer & Yarrow 


Germ Tube - L-Sorbose v L-Arabinose +,w D-Glucitol +,w 

Fermentation Sucrose + D-Arabinose v α-M-D-glucoside + 

Glucose -,w Maltose + D-Ribose v D-Gluconate +,w 

Galactose -,w Cellobiose + L-Rhamnose v DL-Lactate v 

Sucrose -,w Trehalose + D-Glucosamine v myo-Inositol - 

Maltose - Lactose v N-A-D-glucosamine v 2-K-D-gluconate + 

Lactose - Melibiose v Glycerol + D-Glucuronate v 

Trehalose -,w Raffinose + Erythritol v Nitrate - 

Assimilation Melezitose v Ribitol + Urease - 

Glucose + Soluble Starch v Galactitol v 0.1% Cycloheximide v 

Galactose + D-Xylose + D-Mannitol + Growth at 40 O C +,w 

MIC µg/mL 

MIC µg/mL 


Range 

Fluconazole 0.125->64 Amphotericin B 0.06-2 

Itraconazole 0.03->8 Flucytosine 0.06-128 

Posaconazole 0.06-1 Caspofungin 0.06->16 

Voriconazole 0.03-1 Anidulafungin 0.008->16 


Espinel-Ingroff et al. (2001), Pfaller et al. (2003, 2007), Espinel-Ingroff (2003), 

Cuenca-Estrella et al. (2006) and the Australian Candidemia Study.

28 


Synonym: Torulopsis glabrata (Anderson) Lodder & de Vries 


Microscopy: Ovoid to ellipsoidal budding blastoconidia, 3.4 x 2.0 µm in size. No pseudohyphae 

or chlamydospores produced. 


Dalmau Plate Culture on Cornmeal and Tween 80 Agar: Ovoid budding yeast cells 

only. No pseudohyphae produced. 

Key Features: germ tube negative yeast and sugar assimilation pattern. Candida glabrata 

is one of the most common yeast species to be found on the body surface and 

is often isolated as an incidental finding from skin and urine. It has been implicated 

as an “opportunistic” cause of both superficial and systemic infections, especially in 

immunocompromised patients, and it has been isolated from patients with septicemia, 

pyelonephritis, pulmonary infections, endocarditis and hyperalimentation. Approximately 

10% of clinical isolates show azole cross resistance. RG-2 organism. 

Antifungal 

Candida glabrata (Anderson) S.A. Meyer & Yarrow 


Germ Tube - L-Sorbose - L-Arabinose - D-Glucitol - 

Fermentation Sucrose - D-Arabinose - α-M-D-glucoside - 

Glucose + Maltose - D-Ribose - D-Gluconate - 

Galactose - Cellobiose - L-Rhamnose - DL-Lactate - 

Sucrose - Trehalose - D-Glucosamine - myo-Inositol - 

Maltose - Lactose - N-A-D-glucosamine - 2-K-D-gluconate v 

Lactose - Melibiose - Glycerol +,s D-Glucuronate - 


Assimilation Melezitose - Ribitol - Urease - 


Galactose - D-Xylose - D-Mannitol - Growth at 40 O C + 

MIC µg/mL 

MIC µg/mL 


Itraconazole 0.008->16 16 Flucytosine 0.008-16 0.03 

Posaconazole 0.008-8 8 Caspofungin 0.008->8 0.25 

Voriconazole 0.008-16 2 Anidulafungin 0.008-8 nd 



et al. (2006). MIC 90 s from the Australian Candidemia Study (note: in this study 10% 

of primary blood isolates were azole cross-resistant, nd = not done).


Teleomorph: Pichia guilliermondii Wickerham. 

Culture: White to cream-coloured smooth, glabrous yeast-like colonies. 

Microscopy: Spherical to subspherical budding yeast-like cells or blastoconidia, 2.0- 

4.0 x 3.0-6.5 µm. 



with dense verticils of blastoconidia. 

Antifungal 

Candida guilliermondii (Castellani) Langeron & Guerra 


Germ Tube - L-Sorbose v L-Arabinose v D-Glucitol v 

Fermentation Sucrose + D-Arabinose v α-M-D-glucoside v 

Glucose + Maltose + D-Ribose + D-Gluconate v 

Galactose v Cellobiose v L-Rhamnose v DL-Lactate v 

Sucrose + Trehalose + D-Glucosamine + myo-Inositol - 

Maltose - Lactose - N-A-D-glucosamine + 2-K-D-gluconate + 

Lactose - Melibiose v Glycerol + D-Glucuronate - 

Trehalose + Raffinose + Erythritol - Nitrate - 

Assimilation Melezitose v Ribitol + Urease - 

Glucose + Soluble Starch - Galactitol v 0.1% Cycloheximide v 

Galactose + D-Xylose + D-Mannitol v Growth at 37 O C v 


guilliermondii has been isolated from numerous human infections, mostly of cutaneous 

origin. It is also found from normal skin and in sea water, faeces of animals, fig wasps, 

buttermilk, leather, fish, and beer. RG-1 organism. 

MIC µg/mL 

MIC µg/mL 


Itraconazole 0.03-8 1.0 Flucytosine 0.03-8 0.125 

Posaconazole 0.03-8 0.5 Caspofungin 0.125->8 0.5 

Voriconazole 0.03-8 0.25 Anidulafungin 0.06-4 nd 

Good data available. Espinel-Ingroff et al. (2001), Pfaller et al. (2003, 2006, 2007), 

Espinel-Ingroff (2003) and Cuenca-Estrella et al. (2006). MIC 90 s from the Australian 

Candidemia Study (nd = not done).

30 


Candida haemulonii (van Uden & Kolipinski) Meyer & Yarrow 

Synonym: Torulopsis haemulonii van Uden & Kolipinski 


Microscopy: Ovoid to globose, budding yeast-like cells or blastoconidia, 3.0-5.0 x 

3.0-6.5 µm. No pseudohyphae produced. 


Dalmau Plate Culture on Cornmeal and Tween 80 Agar: Spherical to ovoid, budding 

yeast-like cells only. No pseudohyphae produced. 


Germ Tube - L-Sorbose - L-Arabinose -,s D-Glucitol + 

Fermentation Sucrose + D-Arabinose -,s α-M-D-glucoside - 

Glucose + Maltose + D-Ribose -,s D-Gluconate + 

Galactose - Cellobiose - L-Rhamnose +,s DL-Lactate - 

Sucrose + Trehalose + D-Glucosamine +,s myo-Inositol - 

Maltose - Lactose - N-A-D-glucosamine + 2-K-D-gluconate + 

Lactose - Melibiose - Glycerol +,s D-Glucuronate - 

Trehalose +,s Raffinose +,s Erythritol - Nitrate - 

Assimilation Melezitose +,s Ribitol s Urease - 

Glucose + Soluble Starch v Galactitol -,s 0.1% Cycloheximide + 

Galactose -,s D-Xylose -,s D-Mannitol + Growth at 37 O C + 

Key Features: germ tube negative yeast and sugar assimilation pattern. Molecular 

identification may be required. Candida haemulonii has been reported from a few 

cases of fungemia but clinical isolations remain rare. It has also been isolated from 

fish and a dolphin. C. haemulonii may be difficult to distinguish from C. famata using 

some commercial yeast identification systems due to data base limitations. RG-1 organism. 

Antifungal 

MIC µg/mL 

MIC µg/mL 

Range 

Antifungal 

Range 

Fluconazole 32->256 Amphotericin B 2-8 

Itraconazole 0.125-4 Flucytosine 0.008-0.125 

Voriconazole 0.06-0.5 Caspofungin 0.03-0.5 


Rodero et al. (2002) and Khan et al. (2007).


Candida inconspicua (Lodder & Kreger-van Rij) S.A.Meyer & Yarrow 

Synonym: Torulopsis inconspicua Lodder & Kreger-van Rij. 


Microscopy: Ovoidal budding blastoconidia, 2.0-5 x 5.0-11.0 µm. 



yeast cells only. Primitive pseudohyphae may be produced after 14 days. 




Glucose - Maltose - D-Ribose - D-Gluconate - 

Galactose - Cellobiose - L-Rhamnose - DL-Lactate + 

Sucrose - Trehalose - D-Glucosamine + myo-Inositol - 

Maltose - Lactose - N-A-D-glucosamine + 2-K-D-gluconate - 


Trehalose - Raffinose - Erythritol - Nitrate - 





inconspicua is a rare cause of candidemia. RG-1 organism. 

Antifungal 

MIC µg/mL 

MIC µg/mL 


Range 

Fluconazole 4-128 Amphotericin B 0.125->8 

Itraconazole 0.25-8 Flucytosine 1-64 

Posaconazole 0.5-8 Caspofungin 0.008-0.25 

Voriconazole 0.125-4 Anidulafungin nd 


Pfaller et al. (2003), Espinel-Ingroff (2003) and the Australian Candidemia Study (nd 

= not done).

32 


Synonym: Candida pseudotropicalis (Castellani) Basgal. 

Teleomorph: Kluyveromyces marxianus (Hansen) van der Walt. 


Microscopy: Short-ovoid to long-ovoid, budding blastoconidia, 3.0-6.5 x 5.5-11.0 µm, 

sometimes becoming elongate (up to 16.0 µm). 


Dalmau Plate Culture on Cornmeal and Tween 80 Agar: Abundant, long, wavy, 

branched pseudohyphae usually formed, with ovoid blastoconidia, budding off singly, 

in pairs or chains, often in a verticillated position. Note: in some strains pseudohyphae 

may be scarce or almost absent. 


kefyr is a rare cause of candidiasis and is usually associated with superficial cutaneous 

manifestations rather than systemic disease. Environmental isolations have been 

made from cheese and dairy products. RG-1 organism. 

Antifungal 

Candida kefyr (Beijerinck) van Uden & Buckley 



Fermentation Sucrose + D-Arabinose - α-M-D-glucoside - 

Glucose + Maltose - D-Ribose v D-Gluconate - 

Galactose +,s Cellobiose v L-Rhamnose - DL-Lactate + 

Sucrose + Trehalose -,w D-Glucosamine - myo-Inositol - 

Maltose - Lactose v N-A-D-glucosamine - 2-K-D-gluconate - 

Lactose v Melibiose - Glycerol s D-Glucuronate - 

Trehalose - Raffinose + Erythritol - Nitrate - 

Assimilation Melezitose - Ribitol s Urease - 

Glucose + Soluble Starch - Galactitol - 0.1% Cycloheximide + 

Galactose s D-Xylose s D-Mannitol v Growth at 40 O C + 

MIC µg/mL 

MIC µg/mL 

Range MIC90 Antifungal Range MIC90 Fluconazole 0.125-4 0.5 Amphotericin B 0.125->8 1 

Itraconazole 0.03-0.5 0.06 Flucytosine 0.03-64 16 

Posaconazole 0.03-0.25 0.25 Caspofungin 0.03-1 0.125 

Voriconazole 0.008-0.125 0.016 Anidulafungin 0.03-0.5 nd 


Espinel-Ingroff et al. (2001), Pfaller et al. (2003, 2006), Espinel-Ingroff (2003) and 

Cuenca-Estrella et al. (2006). MIC 90 s from the Australian Candidemia Study (nd = 

not done).


Teleomorph: Issatchenkia orientalis Kudryavtesev. 


Microscopy: Predominantly small, elongated to ovoid budding blastoconidia, 2.0-5.5 

x 4.0-15.0 µm. 


Dalmau Plate Culture on Cornmeal and Tween 80 Agar: Abundant long, wavy, 

branched pseudohyphae with elongated to ovoid blastoconidia, budding off in verticillate 

branches. 


krusei is regularly associated with some forms of infant diarrhoea and occasionally 

with systemic disease. It has also been reported to colonise the gastrointestinal, respiratory 

and urinary tracts of patients with granulocytopenia. Environmental isolations 

have been made from beer, milk products, skin, faeces of animals and birds and pickle 

brine. RG-2 organism. 

Antifungal 

Candida krusei (Castellani) Berkhout 




Glucose + Maltose - D-Ribose - D-Gluconate - 







Glucose + Soluble Starch - Galactitol - 0.1% Cycloheximide v 


MIC µg/mL 

MIC µg/mL 

Range MIC90 Antifungal Range MIC90 Fluconazole 0.25->128 64 Amphotericin B 0.06->8 1 

Itraconazole 0.03->8 0.5 Flucytosine 0.5-64 16 

Posaconazole 0.03-1 1 Caspofungin 0.125->4 1 

Voriconazole 0.03-4 0.5 Anidulafungin 0.008-8 nd 




34 


Candida lipolytica (F.C. Harrison) Diddens & Lodder 

Teleomorph: Yarrowia lipolytica (Wickerham et al.) van der Walt & von Arx. 


Microscopy: Spherical, ellipsoidal to elongate budding blastoconidia, 3.0-5 x 3.3-15.0 

µm. 


Dalmau Plate Culture on Cornmeal and Tween 80 Agar: Pseudohyphae and true 

hyphae are produced. 


Germ Tube - L-Sorbose v L-Arabinose - D-Glucitol + 


Glucose - Maltose - D-Ribose v D-Gluconate v 

Galactose - Cellobiose w,- L-Rhamnose - DL-Lactate + 




Trehalose - Raffinose - Erythritol + Nitrate - 

Assimilation Melezitose - Ribitol v Urease - 


Galactose v D-Xylose - D-Mannitol + Growth at 37 O C v 


lipolytica is a rare cause of candidemia. RG-1 organism. 

Antifungal 

MIC µg/mL 

MIC µg/mL 


Range 

Fluconazole 1->64 Amphotericin B 0.06-1 

Itraconazole 0.06-8 Flucytosine 0.125-64 

Posaconazole 0.03-4 Caspofungin 0.25-2 

Voriconazole 0.03-1 Anidulafungin 0.125-0.5 


Espinel-Ingroff et al. (2001), Pfaller et al. (2003), Espinel-Ingroff (2003) and Australian 

Candidemia Study.


Candida lusitaniae van Uden & do Carmo-Sousa 

Teleomorph: Clavispora lusitaniae Rodrigues de Miranda. 


Microscopy: Ovoid to ellipsoidal budding blastoconidia, 1.5-6.0 x 2.5-10.0 µm. 


Dalmau Plate Culture on Cornmeal and Tween 80 Agar: Abundant pseudohyphae 

with short chains of blastoconidia. 


Germ Tube - L-Sorbose + L-Arabinose v D-Glucitol + 

Fermentation Sucrose + D-Arabinose - α-M-D-glucoside v 

Glucose + Maltose + D-Ribose - D-Gluconate s 

Galactose v Cellobiose + L-Rhamnose v DL-Lactate +,w 

Sucrose v Trehalose + D-Glucosamine - myo-Inositol - 

Maltose v Lactose - N-A-D-glucosamine + 2-K-D-gluconate + 



Assimilation Melezitose + Ribitol s Urease - 




lusitaniae is a known cause of disseminated candidiasis, including septicemia and 

pyelonephritis. C. lusitaniae was first isolated from the alimentary tract of warm blooded 

animals and environmental isolations have been made from cornmeal, citrus peel, 

fruit juices, and milk from cows with mastitis. C. lusitaniae may also be difficult to distinguish 

from C. tropicalis using some yeast identification systems. RG-2 organism. 

Antifungal 

MIC µg/mL 

MIC µg/mL 

Range MIC90 Antifungal Range MIC90 Fluconazole 0.125-64 8 Amphotericin B 0.03-8 0.125 

Itraconazole 0.06-2 0.125 Flucytosine 0.03-128 0.03 

Posaconazole 0.008-0.5 0.25 Caspofungin 0.125-4 0.5 

Voriconazole 0.008-2 0.125 Anidulafungin 0.03->8 nd 

Some data available. Espinel-Ingroff et al. (2001), Pfaller et al. (2002b, 2003), 

Espinel-Ingroff (2003) and Cuenca-Estrella et al. (2006). MIC 90 s from the Australian 

Candidemia Study (nd = not done).

36 


Candida norvegensis Dietrichson ex van Uden & Buckley 

Teleomorph: Pichia norvegensis Leask & Yarrow. 


Microscopy: Ovoid, budding blastoconidia, 2.0-3.5 x 3.5-5.0 µm. Pseudohyphae 

rarely produced. 



yeast cells only. No pseudohyphae produced. 




Glucose s Maltose - D-Ribose - D-Gluconate - 

Galactose - Cellobiose + L-Rhamnose - DL-Lactate w 


Maltose - Lactose - N-A-D-glucosamine - 2-K-D-gluconate - 







norvegensis is a very rare clinical isolate that has been reported as a causative agent 

of peritonitis and disseminated candidiasis in a patient on CAPD. RG-1 organism. 

Antifungal 

MIC µg/mL 

MIC µg/mL 

Range 

Antifungal 

Range 

Fluconazole 16 Amphotericin B 1 

Itraconazole 0.25 Flucytosine 8 

Voriconazole 0.125 Posaconazole 0.125 


Pfaller et al. (2002b).


Candida parapsilosis Complex 

Recently Candida parapsilosis has been recognised as 3 species: C. parapsilosis, C. 

orthopsilosis and C. metapsilosis (Tavanti et al. 2005). These three species are phenotypically 

indistinguishable and are best distinguished by genetic analysis. Antifungal 

susceptibility data from the Australian Candidemia Study also shows no significant differences 

between the species. 


Microscopy: Predominantly small, globose to ovoid budding blastoconidia, 2.0-3.5 x 

3.0-4.5 µm, with some larger elongated forms present. 


Dalmau Plate Culture on Cornmeal and Tween 80 Agar: Abundant, much-branched 

pseudohyphae in a delicate tree-like pattern with 2-3 blastoconidia in small clusters at 

intervals along the pseudohyphae. 


Germ Tube - L-Sorbose +,s L-Arabinose + D-Glucitol + 


Glucose + Maltose + D-Ribose v D-Gluconate +,s 

Galactose v Cellobiose - L-Rhamnose - DL-Lactate - 

Sucrose -,s Trehalose + D-Glucosamine v myo-Inositol - 

Maltose -,s Lactose - N-A-D-glucosamine + 2-K-D-gluconate + 


Trehalose -,s Raffinose - Erythritol - Nitrate - 

Assimilation Melezitose + Ribitol +,s Urease - 




parapsilosis is an opportunistic human pathogen which may cause cutaneous infections, 

especially of the nail and systemic disease, especially endocarditis. Other clinical 

manifestations include endophthalmitis and fungemia. Environmental isolations have 

been made from intertidal and oceanic waters, pickle brine, cured meats, olives and 

normal skin, and faeces. RG-1 organism. 

Antifungal 

MIC µg/mL 

MIC µg/mL 


Itraconazole 0.015-2 0.25 Flucytosine 0.03->64 0.25 

Posaconazole 0.008-0.5 0.03 Caspofungin 0.03->8 1 

Voriconazole 0.008-2 0.25 Anidulafungin 0.008->8 nd 




38 


Candida pelliculosa Redaelli 

Teleomorph: Pichia anomala (Hansen) Kurtzman. 


Microscopy: Spherical to ellipsoidal budding blastoconidia, 2-4 x 2-6 µm. Pseudohyphae 

may be present. Asci when present, containing 1-4 hat-shaped ascospores. 


Dalmau Plate Culture on Cornmeal and Tween 80 Agar: Spherical to ellipsoidal 

budding yeast cells and abundant pseudohyphae in most strains. 


Germ Tube - L-Sorbose - L-Arabinose v D-Glucitol + 


Glucose + Maltose + D-Ribose v D-Gluconate v 

Galactose v Cellobiose + L-Rhamnose - DL-Lactate + 


Maltose v Lactose - N-A-D-glucosamine - 2-K-D-gluconate - 

Lactose - Melibiose - Glycerol + D-Glucuronate n 

Trehalose - Raffinose + Erythritol + Nitrate + 

Assimilation Melezitose + Ribitol v Urease - 

Glucose + Soluble Starch + Galactitol - 0.1% Cycloheximide - 

Galactose v D-Xylose v D-Mannitol + Growth at 37 O C v 


pelliculosa has been reported from cases of candidemia and catheter related infections 

in humans and has been isolated from soil, grains, fruit and warm blooded animals. 


Antifungal 

MIC µg/mL 

MIC µg/mL 

Range 

Antifungal 

Range 



Posaconazole 0.125-1 Caspofungin 0.06-0.5 

Voriconazole 0.06-0.25 Anidulafungin nd 

Limited data available, antifungal susceptibility testing of individual strains is 

recommended. Pfaller et al. (2003), Espinel-Ingroff (2003), Cuenca-Estrella et al. 

(2006) and the Australian Candidemia Study (nd = not done).


Candida rugosa (Anderson) Diddens & Lodder 


Microscopy: Ellipsoidal to elongate budding blastoconidia, 6-10 x 2-3.5 µm. Sometimes 

short pseudohyphae may be produced. 


Dalmau Plate Culture on Cornmeal and Tween 80 Agar: Ellipsoidal budding yeast 

cells only and densely branched pseudohyphae produced. 


Germ Tube - L-Sorbose v L-Arabinose - D-Glucitol +,s 


Glucose - Maltose - D-Ribose - D-Gluconate v 

Galactose - Cellobiose - L-Rhamnose - DL-Lactate +,s 


Maltose - Lactose - N-A-D-glucosamine s 2-K-D-gluconate - 



Assimilation Melezitose - Ribitol -,s Urease - 


Galactose + D-Xylose v D-Mannitol +,s Growth at 37 O C + 


rugosa has been associated with catheter related fungemia and has been isolated 

from human and bovine faeces, sea water and soil. RG-1 organism. 

Antifungal 

MIC µg/mL 

MIC µg/mL 


Range 



Posaconazole 0.06-0.25 Caspofungin 0.25-2 

Voriconazole 0.008-0.25 Anidulafungin 0.03-4 


Espinel-Ingroff et al. (2001), Pfaller et al. (2003), Espinel-Ingroff (2003) and the 

Australian Candidemia Study.

40 



Microscopy: Spherical to subspherical budding yeast-like cells or blastoconidia, 3-5.5 

x 4-9 µm. 


Dalmau Plate Culture on Cornmeal and Tween 80 Agar: Abundant, long, wavy, 

branched pseudohyphae with numerous ovoid blastoconidia, budding off. Terminal 

vesicles (chlamydospores) are not produced. 


tropicalis is a major cause of septicemia and disseminated candidiasis. It is also found 

as part of the normal human mucocutaneous flora and environmental isolations have 

been made from faeces, shrimp, kefir, and soil. RG-2 organism. 

Antifungal 

Candida tropicalis (Castellani) Berkhout 


Germ Tube - L-Sorbose v L-Arabinose - D-Glucitol + 

Fermentation Sucrose v D-Arabinose - α-M-D-glucoside v 

Glucose + Maltose + D-Ribose -,s D-Gluconate v 

Galactose + Cellobiose +,s L-Rhamnose - DL-Lactate v 

Sucrose v Trehalose + D-Glucosamine v myo-Inositol - 



Trehalose +,s Raffinose - Erythritol - Nitrate - 

Assimilation Melezitose v Ribitol +,s Urease - 



MIC µg/mL 

MIC µg/mL 


Itraconazole 0.03->8 0.5 Flucytosine 0.03->64 0.125 

Posaconazole 0.008->8 0.06 Caspofungin 0.03->8 0.25 

Voriconazole 0.008->8 0.25 Anidulafungin 0.03->8 nd 





Chaetomium Kunze ex Fries 

This genus is a common ascomycete (pyrenomycete) characterised by the formation of 

darkly-pigmented, globose, ovoid, barrel to flask-shaped, ostiolate ascocarps (perithecia) 

beset with dark-coloured terminal hairs (setae) which are straight, branched or 

curved. Asci are clavate to cylindrical, typically eight-spored and evanescent. Ascospores 

are one-celled, darkly-pigmented, smooth-walled, of varying shape, mostly 

ovoid, ellipsoidal or lemon-shaped. Chlamydospores and solitary conidia may also be 

produced. RG-1 organism. 

The genus contains between 160 and 180 species, and all are saprophytic being isolated 

from soil, straw, dung and plant debris. However, quite a few species are thermophilic 

and can grow at temperatures above 37 O C. Chaetomium species are important 

agents for the decomposition of cellulose waste and plant materials, and are only 

rarely isolated in medical mycology laboratories. 

Key Features: ascomycete producing darkly-pigmented ostiolate perithecia beset with 

long dark terminal setae. 

For descriptions of species, keys to taxa and additional information see Ames (1963), 

Seth (1970), Millner (1975), Domsch et al. (1980), Ellis and Keane (1981), Ellis (1981) 

and de Hoog et al. (2000). 

100 μm 

10 μm 

Ascocarp (perithecia), terminal hairs, asci and ascospores of Chaetomium. 

Antifungal 






McGinnis and Pasarell (1998) and WCH in-house data.

42 


Chrysosporium Corda 

Colonies are moderately fast growing, flat, white to tan to beige in colour, often with a 

powdery or granular surface texture. Reverse pigment absent or pale brownish-yellow 

with age. Hyaline, one-celled conidia are produced directly on vegetative hyphae 

by non-specialised conidiogenous cells. Conidia are typically pyriform to clavate with 

truncate bases and are formed either intercalary (arthroconidia), laterally (often on 

pedicels) or terminally. 

Species of Chrysosporium are occasionally isolated from skin and nail scrapings, especially 

from feet, but because they are common soil saprophytes they are usually 

considered as contaminants. There are about 22 species of Chrysosporium, several 

are keratinolytic with some also being thermotolerant, and cultures may closely resemble 

some dermatophytes, especially Trichophyton mentagrophytes, and some strains 

may also resemble cultures of Histoplasma and Blastomyces. 

Chrysosporium tropicum Carmichael 

Colonies are flat, white to cream-coloured with a very granular surface. Reverse pigment 

absent or pale brownish-yellow with age. Microscopically, conidia are numerous, 

hyaline, single-celled, clavate to pyriform, smooth, slightly thick-walled (6-7 x 3.5-4 

µm), and have broad truncate bases and pronounced basal scars. The conidia are 

formed at the tips of the hyphae, on short or long lateral branches, or sessile along the 

hyphae (intercalary). No macroconidia or hyphal spirals are seen. RG-2 organism. 

For descriptions of species, keys to taxa and additional information see Carmichael 

(1962), Rebell and Taplin (1970), Sigler and Carmichael (1976), Van Oorschot (1980), 

Domsch et al. (1980) and de Hoog et al. (2000). 

10 μm 

Chrysosporium tropicum showing typical pyriform to clavate-shaped conidia 

with truncated bases which may be formed either intercalary, laterally 

or terminally.


Cladophialophora bantiana (Saccardo) de Hoog et al. 

Synonym: Xylohypha bantiana (Saccardo) McGinnis, Borelli and Ajello 

Colonies are moderately fast growing, olivaceous-grey, suede-like to floccose and 

grow at temperatures up to 42-43 O C. Conidia are formed in long, sparsely branched, 

flexuose, acropetal chains from undifferentiated conidiophores. Conidia are one-celled 

(very occasionally two-celled), pale brown, smooth-walled, ellipsoid to oblong-ellipsoid 

and are 2-3 x 4-7 µm in size. 

Cladophialophora bantiana has been isolated from soil and is a recognised agent of 

cerebral phaeohyphomycosis. The fungus is neurotropic and may cause brain abscesses 

in both normal and immunosuppressed patients. RG-2 organism. 

Cladophialophora bantiana may be distinguished from Cladosporium species by the 

absence of conidia with distinctly pigmented hila, the absence of characteristic shield 

cells and by growth at 42 O C (compared with Cladophialophora carrionii which has a 

maximum growth temperature of 35-36 O C, and Cladosporium species which have a 

maximum of less than 35 O C). 


(1980), McGinnis and Borelli (1981), McGinnis et al. (1986a), Rippon (1988), Kwon- 

Chung and Bennett (1992) and de Hoog et al. (2000). 

10 μm 

Conidiophore and conidia of Cladophialophora bantiana. 

Antifungal 

MIC µg/mL 


MIC µg/mL 

Range MIC90 Amphotericin B 0.03-2 0.5 Posaconazole 0.008-0.06 0.06 

Itraconazole 0.03-0.5 0.5 Voriconazole 0.03-1 0.125 


McGinnis and Pasarell (1998), Espinel-Ingroff et al. (2001), Espinel-Ingroff (2001) 

and WCH in-house data.

44 


Cladophialophora carrionii (Trejos) de Hoog et al. 

Synonym: Cladosporium carrionii Trejos 

Colonies are slow growing, reaching 3-4 cm in diameter after one month, with a compact 

suede-like to downy surface and are olivaceous-black in colour. Microscopy shows 

ascending to erect, olivaceous-green, apically branched, elongate conidiophores 

producing branched acropetal chains of smooth-walled conidia. Conidia are pale 

olivaceous, smooth-walled or slightly verrucose, limoniform to fusiform, 1.5-3.0 x 2.0- 

7.0 µm in size. Bulbous phialides with large collarettes and minute, hyaline conidia 

are occasionally formed on nutritionally poor media. Maximum growth temperature 

35-37 O C. RG-2 organism. 

Cladophialophora carrionii is a recognised agent of chromoblastomycosis and it 

has been isolated from soil and fence posts made from Eucalyptus sp. Cases of 

chromoblastomycosis caused by C. carrionii are commonly found in Australia, 

Venezuela, Madagascar and South America. Isolates from phaeomycotic cysts and 

opportunistic infections have also been reported. 

Key Features: conidia are smaller and comprise heavily branched systems which fall 

apart much more easily than in the other Cladophialophora species. 


(1980), Rippon (1988), de Hoog et al. (1995) and de Hoog et al. (2000). 

10 μm 

Conidiophores and conidia of Cladophialophora carrionii. 

Antifungal 

MIC µg/mL 


MIC µg/mL 

Range MIC90 Amphotericin B 0.06-4 1 Posaconazole 0.06-0.5 0.25 

Itraconazole 0.03-0.5 0.5 Voriconazole 0.03-0.5 0.25 


McGinnis and Pasarell (1998), Espinel-Ingroff et al. (2001), Gonzales et al. (2005) 



Cladosporium Link ex Fries 

Colonies are rather slow growing, mostly olivaceous-brown to blackish brown but also 

sometimes grey, buff or brown, suede-like to floccose, often becoming powdery due 

to the production of abundant conidia. Vegetative hyphae, conidiophores and conidia 

are equally pigmented. Conidiophores are more or less distinct from the vegetative 

hyphae, being erect, straight or flexuose, unbranched or branched only in the apical 

region, with geniculate sympodial elongation in some species. Conidia are produced in 

branched acropetal chains, being smooth, verrucose or echinulate, one- to four-celled, 

and have a distinct dark hilum. The term blastocatenate is often used to describe 

chains of conidia where the youngest conidium is at the apical or distal end of the chain. 

Note: the conidia closest to the conidiophore, and where the chains branch, are usually 

“shield-shaped”. The presence of shield-shaped conidia, a distinct hilum, and chains 

of conidia that readily disarticulate, are diagnostic for the genus Cladosporium. 

Cladosporium species have a world-wide distribution and are amongst the most common 

of air-borne fungi. Some 500 species have been described. Isolates of Cladosporium 

are frequently isolated as contaminants. RG-1 organisms. The pathogenic species 

have now been transferred to the genus Cladophialophora. 

Key Features: dematiaceous hyphomycete forming branched acropetal chains of 

conidia, each with a distinct hilum. 

For descriptions of species, keys to taxa and additional information see Ellis (1971 and 

1976), Domsch et al. (1980), McGinnis (1980) and de Hoog et al. (2000). 

Antifungal 

10 μm 

Conidiophores and conidia of Cladosporium cladosporioides. 




Amphotericin B 0.03-8 Itraconazole 0.03-32 Voriconazole 0.06-1 



46 


Coccidioides immitis/posadasii complex 

Recently Coccidioides immitis has been recognised as 2 species: C. immitis and C. 

posadasii (Fisher et al. 2002). The two species are morphologically identical and can 

be distinguished only by genetic analysis and different rates of growth in the presence 

of high salt concentrations (C. posadasii grows more slowly). C. immitis is geographically 

limited to California’s San Joaquin Valley region, whereas C. posadasii is found in 

the desert regions of the USA southwest, Mexico and South America. The two species 

appear to coexist in the desert regions of the USA southwest and Mexico. 

Colonies of C. immitis/posadasii on Sabouraud’s dextrose agar at 25 O C are initially 

moist and glabrous, but rapidly become suede-like to downy, greyish white with a tan 

to brown reverse, however considerable variation in growth rate and culture morphology 

has been noted. Microscopy shows typical single-celled, hyaline, rectangular to 

barrel-shaped, alternate arthroconidia, 2.5-4 x 3-6 µm in size, separated from each 

other by a disjunctor cell. This arthroconidial state has been classified in the genus 

Malbranchea and is similar to that produced by many non-pathogenic soil fungi, e.g. 

Gymnoascus species. 

WARNING: RG-3 organism. Cultures of Coccidioides immitis/posadasii represent a 

severe biohazard to laboratory personnel and must be handled with extreme caution in 

an appropriate pathogen handling cabinet. C. immitis/posadasii is a dimorphic fungus, 

existing in living tissue as spherules and endospores, and in soil or culture in a mycelial 

form. Culture identification by exoantigen test is now the method of choice. 

Key Features: clinical history, tissue pathology, culture identification by positive 


20 μm 

Tissue morphology showing typical endosporulating spherules of C. immitis. Young 

spherules have a clear centre with peripheral cytoplasm and a prominent thick-wall. 

Endospores (sporangiospores) are later formed within the spherule by repeated cytoplasmic 

cleavage. Rupture of the spherule releases endospores into the surrounding 

tissue where they re-initiate the cycle of spherule development.


Coccidioides immitis/posadasii complex 

5 μm 

Culture and arthroconidia separated from each other by disjunctor 

cells of Coccidioides immitis/posadasii. 

For descriptions of species, keys to taxa and additional information see Ajello (1957), 

Steele et al. (1977), McGinnis (1980), Chandler et al. (1980), Catanzaro (1986), Rippon 

(1988), de Hoog et al. (2000) and Fisher et al. (2002). 

Antifungal 

MIC µg/mL 


MIC µg/mL 

Range MIC90 Amphotericin B 0.06-2 1 Posaconazole 0.03-1 0.5 

Fluconazole 2-64 32 Voriconazole 0.03-1 0.5 


Limited data available. Espinel-Ingroff et al. (2001), Espinel-Ingroff (2003), Gonzalez 

et al. (2005) and Sabatelli et al. (2006).

48 


Colletotrichum coccodes (Wallroth) S. Hughes 

Colonies usually darkly pigmented with white aerial mycelium, consisting of numerous 

black sclerotia and light brown-coloured conidial masses, reverse is dark brown. 

Sclerotia are usually abundant, setose, spherical and are often confluent. Conidia are 

straight, fusiform, attenuated at the ends, 16-22 x 3-4 µm. Appressoria are common, 

clavate, brown, 11-16.5 x 6-9.5 µm, variable in shape. RG-1 organism. 

Over 500 Colletotrichum species have been reported. C. coccodes is a common soil 

and plant pathogen widely distributed in Africa, Asia, Australasia, Europe, and the 

Americas. It has been reported from a case of human mycotic keratitis. 


(1980), McGinnis (1980) and de Hoog et al. (2000). 

20 μm 

10 μm 

50 μm 

Sclerotia with setae, conidia and appressoria of C. coccodes.


Synonym: Entomophthora coronata (Costantin) Kevorkian 

The species of the genus Conidiobolus produce characteristic multinucleate primary 

and secondary (replicative) conidia on top of unbranched conidiophores. Each subspherical 

conidium is discharged as a result of the pressure developed within the conidium, 

and each bears a more or less prominent papilla after discharge (King 1983). 

The genus contains 27 species, however C. coronatus and C. incongruus are the only 

species that are known to cause human disease, although C. lamprauges has also 

been reported once from a horse (Humber, Brown and Kornegay, 1989). 

Colonies of C. coronatus grow rapidly and are flat, cream-coloured, glabrous becoming 

radially folded and covered by a fine, powdery, white surface mycelium and conidiophores. 

The lid of the petri dish soon becomes covered with conidia, which are 

forcibly discharged by the conidiophores. The colour of the colony may become tan to 

brown with age. Conidiophores are simple forming solitary, terminal conidia which are 

spherical, 10 to 25 µm in diameter, single-celled and have a prominent papilla. Conidia 

may also produce hair-like appendages, called villae. Conidia germinate to produce 

either, (1) single or multiple hyphal tubes that may also become conidiophores which 

bear secondary conidia, or (2) replicate by producing multiple short conidiophores, 

each bearing a small secondary conidium. RG-2 organism. 

Conidiobolus coronatus is commonly present in soil and decaying leaves. It has a 

world-wide distribution especially tropical rain forests of Africa. Human infections are 

usually restricted to the rhinofacial area. However, there are occasional reports of 

dissemination to other sites. All human infections have been confined to the tropics. 

For descriptions of species, keys to taxa and additional information see Emmons and 

Bridges (1961), King (1976a, 1976b, 1983), McGinnis (1980), Rippon (1988), Kwon- 

Chung and Bennett (1992), de Hoog et al. (2000) and Ellis (2005a). 

10 μm 

Conidiobolus coronatus (Costantin) Batko 

10 μm 

Spherical conidia with hair-like appendages (villae) and prominent papillae 

characteristic of Conidiobolus coronatus.

50 


Cryptococcus Kützing emend. Phaff & Spencer 

The genus Cryptococcus is characterised by globose to elongate yeast-like cells or 

blastoconidia that reproduce by multilateral budding, by polar budding on a narrow 

base (may or may not be present), and pseudohyphae being absent or rudimentary. 

Most strains have capsulated cells; extent of capsule formation depends on the medium. 

Under certain conditions of growth the capsule may contain starch-like compounds, 

which are released into the medium by many strains. On solid media the 

cultures are generally mucoid or slimy in appearance; red, orange or yellow carotenoid 

pigments may be produced, but young colonies of most species are usually non-pigmented, 

being cream in colour. No fermentation: Nitrate assimilated or not: Inositol 

assimilated. The genus Cryptococcus is similar to the genus Rhodotorula. The distinctive 

difference between the two is the assimilation of inositol, which is positive in 

Cryptococcus. 

Cryptococcosis is a chronic, subacute to acute pulmonary, systemic or meningitic disease, 

initiated by the inhalation of basidiospores and/or desiccated yeast cells of Cryptococcus 

species. Primary pulmonary infections have no diagnostic symptoms and 

are usually subclinical. On dissemination, the fungus usually shows a predilection for 

the central nervous system, however skin, bones and other visceral organs may also 

become involved. Although C. neoformans and C. gattii are regarded as the principle 

pathogenic species, Cryptococcus albidus and Cryptococcus laurentii have on occasion 

also been implicated in human infection. 

5 μm 

Culture appearances on Bird Seed Agar of Cryptococcus neoformans (brown 

colonies) and Candida albicans (white colonies) and India ink preparation of 

C. neoformans surrounded by a characteristic wide gelatinous capsule. 

For descriptions of species, keys to taxa and additional information see Rippon (1982), 

Barnett et al. (1983), McGinnis (1980), Kurtzman and Fell (1988), Casadevall and Perfect 

(1988) and de Hoog et al. (2000).


Cryptococcus albidus (Saito) Skinner 

Culture: Colonies (SDA) are cream-coloured smooth, mucoid glabrous yeast-like. 

Budding yeast cells only. No pseudohyphae present. 

Microscopy: Globose to ovoid budding yeast-like cells, 3.5-8.8 x 5.5-10.2 μm. 

India Ink Preparation: Positive - Distinct capsules are present. 


Germ Tube - L-Sorbose v L-Arabinose + D-Glucitol + 


Glucose - Maltose + D-Ribose v D-Gluconate + 

Galactose - Cellobiose + L-Rhamnose v DL-Lactate v 

Sucrose - Trehalose +,w D-Glucosamine - myo-Inositol + 

Maltose - Lactose v N-A-D-glucosamine - 2-K-D-gluconate + 

Lactose - Melibiose v Glycerol v D-Glucuronate + 

Trehalose - Raffinose + Erythritol v Nitrate + 

Assimilation Melezitose + Ribitol v Urease + 

Glucose + Soluble Starch v Galactitol v 0.1% Cycloheximide - 

Galactose v D-Xylose + D-Mannitol + Growth at 37 O C v 

C. albidus has variable growth at 37 O C, and rare human infections have been reported 

however its pathogenicity is questionable. RG-1 organism. 

Cryptococcus laurentii (Kufferath) Skinner 

Culture: Colonies (SDA) are cream-coloured, often becoming a deeper orange-yellow 

with age, with a smooth mucoid texture. Budding yeast cells only. No pseudohyphae 

present. 

Microscopy: Spherical and elongated budding yeast-like cells or blastoconidia, 2.0- 

5.5 x 3.0-7.0 μm. No pseudohyphae present. 

India Ink Preparation: Positive - Narrow but distinct capsules are present. 


Germ Tube - L-Sorbose v L-Arabinose + D-Glucitol + 

Fermentation Sucrose + D-Arabinose + α-M-D-glucoside + 

Glucose - Maltose + D-Ribose + D-Gluconate + 

Galactose - Cellobiose + L-Rhamnose + DL-Lactate v 

Sucrose - Trehalose + D-Glucosamine - myo-Inositol + 

Maltose - Lactose + N-A-D-glucosamine - 2-K-D-gluconate + 

Lactose - Melibiose + Glycerol v D-Glucuronate + 

Trehalose - Raffinose + Erythritol v Nitrate - 

Assimilation Melezitose + Ribitol + Urease + 

Glucose + Soluble Starch v Galactitol + 0.1% Cycloheximide - 

Galactose v D-Xylose + D-Mannitol + Growth at 37 O C -,w 

Note: some strains of C. laurentii may develop a brown pigment on Bird Seed agar and 

turn CGB media blue, similar to C. gattii, however C. laurentii assimilates both lactose 

and melibiose while C. gattii does not. Rare human infections have been reported 

however its pathogenicity is questionable. RG-1 organism.

52 


Cryptococcus gattii (Vanbreus. & Takashio) Kwon-Chung & Boekhout 

Teleomorph: Filobasidiella bacillispora Kwon-Chung, 

Synonym: Cryptococcus neoformans var. gattii Vanbreus & Takashio 

Cryptococcus gattii has two serotypes (B and C) and has recently been reclassified as 

a separate species (Kwon-Chung et al. 2002). C. gattii generally has a more restricted 

geographical distribution than C. neoformans, causing human disease in climates 

ranging from temperate to tropical Australia, Papua New Guinea, parts of Africa, India, 

southeast Asia, Mexico, Brazil, Paraguay and Southern California, although recent 

infections have also been reported from Vancouver Island, Canada. Non-immunocompromised 

hosts are usually affected and large mass lesions in the lung and/or brain 

(cryptococcomas) are often present (Sorrell, 2001). 

Canavanine glycine bromothymol blue (CGB) agar (Kwon-Chung et al. 1982) is the 

media of choice to differentiate C. gattii from C. neoformans. This simple biotype test 

is based on the ability of C. gattii isolates to grow in the presence of L-canavanine and 

to assimilate glycine as a sole carbon source. 

C. gattii turns CGB agar blue within 2-5 days; 

C. neoformans does not grow on this medium 

Culture: Colonies (SDA) cream-coloured smooth, mucoid yeast-like colonies. 

Microscopy: Globose to ovoid budding yeast-like cells 3.0-7.0 x 3.3- 7.9 µm. 

India Ink Preparation: Positive - Distinct, wide gelatinous capsules are present. 


No pseudohyphae present. 

Bird Seed Agar: Colonies turn dark brown in colour as they selectively absorb a 

brown pigment from this media. Colonies are often more mucoid when compared to 

C. neoformans (Staib, 1987). 

Canavanine-Glycine-Bromothymol Blue (CGB) Agar: turns blue within 2-5 days. 

Key Features: encapsulated yeast; absence of pseudohyphae; growth at 37 O C; positive 

hydrolysis of urea; negative fermentation of sugars and positive assimilation of 

glucose, maltose, sucrose, galactose, trehalose, raffinose, inositol, cellobiose, rhamnose, 

arabinose, melezitose and xylose, and negative assimilation of nitrate, lactose, 

melibiose, erythritol and soluble starch; growth on bird seed (Guizotia abyssinica seed) 

or caffeic acid agar - colonies turn a dark brown colour; growth on CGB agar turning it 

blue within 2-5 days. RG-2 organism, however mating experiments for the production 

of basidiospores should be done in an appropriate pathogen handling cabinet.


Cryptococcus gattii (Vanbreus. & Takashio) Kwon-Chung & Boekhout 


Germ Tube - L-Sorbose v L-Arabinose +,w D-Glucitol + 



Galactose - Cellobiose +,w L-Rhamnose + DL-Lactate - 

Sucrose - Trehalose + D-Glucosamine v myo-Inositol + 

Maltose - Lactose - N-A-D-glucosamine v 2-K-D-gluconate + 

Lactose - Melibiose - Glycerol - D-Glucuronate + 

Trehalose - Raffinose +,w Erythritol - Nitrate - 


Glucose + Soluble Starch + Galactitol + 0.1% Cycloheximide - 


Antifungal 

MIC µg/mL 

MIC µg/mL 

Range MIC90 Antifungal Range MIC90 Fluconazole 1->64 16 Amphotericin B 0.03-2 0.25 

Itraconazole

54 


Cryptococcus neoformans (Sanfelice) Vuillemin 

Culture: Colonies (SDA) cream-coloured smooth, mucoid yeast-like colonies. 

Microscopy: Globose to ovoid budding yeast-like cells 3.0-7.0 x 3.3- 7.9 µm. 

India Ink Preparation: Positive - Distinct, wide gelatinous capsules are present. 


No pseudohyphae present. 

Bird Seed Agar: Colonies turn dark brown in colour as colonies selectively absorb a 

brown pigment from this media (Staib, 1987). 

Canavanine-Glycine-Bromothymol Blue (CGB) Agar: leaves this medium unchanged. 

Creatinine dextrose bromothymol blue thymine (CDBT) agar: Cryptococcus neoformans 

var. neoformans grows as bright red colonies, turning the medium a bright 

orange after 5 days. No colour change is observed for C. neoformans var. grubii. 


Germ Tube - L-Sorbose v L-Arabinose +,w D-Glucitol + 



Galactose - Cellobiose +,w L-Rhamnose + DL-Lactate - 

Sucrose - Trehalose + D-Glucosamine v myo-Inositol + 

Maltose - Lactose - N-A-D-glucosamine v 2-K-D-gluconate + 

Lactose - Melibiose - Glycerol - D-Glucuronate + 

Trehalose - Raffinose +,w Erythritol - Nitrate - 


Glucose + Soluble Starch + Galactitol + 0.1% Cycloheximide - 


Key Features: encapsulated yeast; absence of pseudohyphae; growth at 37 O C; positive 

hydrolysis of urea; negative fermentation of sugars and positive assimilation of 

glucose, maltose, sucrose, galactose, trehalose, raffinose, inositol, cellobiose, rhamnose, 

arabinose, melezitose and xylose, and negative assimilation of nitrate, lactose, 

melibiose, erythritol and soluble starch; growth on bird seed (Guizotia abyssinica seed) 

or caffeic acid agar - colonies turn a dark brown colour; does not growth on CGB agar 

(no colour change). RG-2 organism, however mating experiments for the production 

of basidiospores should be done in an appropriate pathogen handling cabinet. 

Antifungal 

MIC µg/mL 

MIC µg/mL 

Range MIC90 Antifungal Range MIC90 Fluconazole 1->64 8 Amphotericin B 0.03-2 0.5 

Itraconazole


Cunninghamella bertholletiae Stadel 

Synonyms: Cunninghamella elegans Lendner 

Cunninghamella echinulata var. elegans (Lendner) Lunn & Shipton 

The genus Cunninghamella is characterised by white to grey, rapidly growing colonies, 

producing erect, straight, branching sporangiophores. These sporangiosphores 

end in globose or pyriform-shaped vesicles from which several one-celled, globose to 

ovoid, echinulate or smooth-walled sporangiola develop on swollen denticles. Chlamydospores 

and zygospores may also be present. 

Colonies are very fast growing, white at first, but becoming rather dark grey and powdery 

with sporangiola development. Sporangiophores to 20 μm wide, straight, with 

verticillate or solitary branches. Vesicles subglobose to pyriform, the terminal ones up 

to 40 µm and the lateral ones 10-30 µm in diameter. Sporangiola are globose (7-11 

µm diameter), or ellipsoidal (9-13 x 6-10 μm), verrucose or short-echinulate, hyaline 

singly but brownish in mass. Temperature: optimum 25 to 30 O C: maximum up to 50 O C. 


Cunninghamella species are mainly soil fungi of the Mediterranean and subtropical 

zones; they are only rarely isolated in temperate regions. The genus now contains 

seven species with C. bertholletiae the only known species to cause disease in man 

and animals which is often associated with trauma and immunosuppression. 

Key Features: zygomycete, clinical isolates grow at 40 O C, one-celled, globose to 

ovoid, echinulate sporangiola borne on swollen terminal or lateral globose to clavate 

fertile vesicles. 

Once again, there has been some confusion as to the correct name of this zygomycete. 

Many medical mycologists (McGinnis, 1980, Weitzman, 1984, and Rippon, 1988) 

preferred the name Cunninghamella bertholletiae because of the thermophilic nature 

of human isolates that grow at temperatures as high as 45 O C. However, Samson 

(1969) and Domsch et al. (1980) preferred the name Cunninghamella elegans and 

Lunn and Shipton (1983) went further and reduced C. elegans (= C. bertholletiae) to 

a variety of Cunninghamella echinulata; i.e. C. echinulata var. elegans. However, C. 

bertholletiae is currently the most acceptable name; C. elegans differs by having purely 

grey colonies and by not growing at temperatures above 40 O C (Weitzman and Crist, 

1979 and de Hoog et al. 2000). 

Antifungal 

MIC µg/mL 

MIC µg/mL 

Antifungal 


Fluconazole >64 >64 Amphotericin B 0.125-8 2 


Posaconazole 0.003-1 1 Voriconazole 8->64 >64 


Sun et al. (2002), Dannaoui et al. (2003), Espinel-Ingroff (2001, 2003), Singh et al. 

(2005), Sabatelli et al. (2006) and WCH in-house data.

56 


10 μm 

Cunninghamella bertholletiae Stadel 

10 μm 

Microscopic morphology of Cunninghamella bertholletiae showing simple sporangiophores 

forming a swollen, terminal vesicle around which single-celled, globose 

to ovoid sporangiola develop on swollen denticles. 


(1980), Weitzman (1984), Lunn and Shipton (1983), Domsch et al. (1980), Samson 

(1969), de Hoog et al (2000) and Ellis (2005b).


Teleomorph: Cochliobolus Drechsler 

Curvularia Boedijn 

Colonies are fast growing, suede-like to downy, brown to blackish brown with a black 

reverse. Conidia are pale brown, with three or more transverse septa (phragmoconidia) 

and are formed apically through a pore (poroconidia) in a sympodially elongating 

geniculate conidiophore similar to Drechslera. Conidia are cylindrical or slightly 

curved, with one of the central cells being larger and darker, germination is bipolar and 

some species may have a prominent hilum. 

The genus Curvularia contains some 35 species which are mostly subtropical and 

tropical plant parasites; however three ubiquitous species, C. lunata, C. pallescens 

and C. geniculata have been recovered from human infections, principally from cases 

of mycotic keratitis. However, cases of subcutaneous, sinusitis, endocarditis, peritonitis 

and disseminated infection have also been reported in immunosuppressed patients. 

RG-1 organisms. 

Key Features: dematiaceous hyphomycete producing sympodial, pale brown, 

cylindrical or slightly curved phragmoconidia, with one of the central cells being larger 

and darker. 

For descriptions of species, keys to taxa and additional information see Ellis (1971), 

Domsch et al. (1980), McGinnis (1980), Rippon (1988) and de Hoog et al. (2000). 

Antifungal 

20 µm 

Conidia of Curvularia lunata. 







58 


Teleomorph: Nectria (Fries) Fr. 

Cylindrocarpon Wollenw. 

Colonies are fast growing, hyaline or bright-coloured, suede-like or woolly. Sporodochia 

may occasionally be present. Conidiophores consist of simple or repeatedly verticillate 

phialides, arranged in brush-like structures. Phialides are cylindrical to subulate, 

with small collarettes producing hyaline, smooth-walled conidia, which are arranged in 

slimy masses. Two types of conidia may be produced; macroconidia which are one to 

several septate, hyaline, straight or curved, cylindrical to fusiform, with a rounded apex 

and flat base; and microconidia which are one-celled, which are usually clearly distinct 

from the macroconidia. Chlamydospores may be present or absent, hyaline to brown, 

spherical, formed singly, in chains or in clumps, intercalary or terminal. RG-2 organism 

if isolated from humans. 

The genus contains 35 species, is widespread, isolated mostly from soil and is recorded 

as an occasional human and animal pathogen. Cylindrocarpon differs from 

Fusarium by lacking an asymmetrical foot-cell on the macroconidia. 

For descriptions of species, keys to taxa and additional information see Booth (1966), 

Domsch et al. (1980) and de Hoog et al. (2000). 

15 µm 

15 µm 

Culture, chlamydospores and macroconidia of Cylindrocarpon lichenicola.


Teleomorph: Pyrenophora Fries 

Drechslera Ito 

Colonies are fast growing, suede-like to downy, brown to blackish brown with a black 

reverse. Conidia are pale to dark brown, usually cylindrical or subcylindrical, straight, 

smooth-walled, and are formed apically through a pore (poroconidia) on a sympodially 

elongating, geniculate conidiophore. Conidia are transversely septate (phragmoconidia), 

with the septum delimiting the basal cell formed first during conidium maturation. 

Germinating is from any or all cells and the hilum is not protuberant. RG-1 organism. 

McGinnis et al. (1986b) have reviewed the isolates from human and animal disease 

purported to be Drechslera or Helminthosporium and concluded that all pathogenic 

isolates examined actually belong to the genera Bipolaris or Exserohilum. 

Key Features: dematiaceous hyphomycete producing sympodial, pale brown, 

cylindrical or subcylindrical, transversely septate poroconidia. 

For descriptions of species, keys to taxa and additional information see Luttrell (1978), 

Ellis (1971 and 1976), Domsch et al. (1980), McGinnis (1980), McGinnis et al. (1986b), 

Sivanesan (1987), Rippon (1988) and de Hoog et al. (2000). Also see Descriptions for 

Bipolaris, Curvularia and Exserohilum. 

Antifungal 

Conidia of Drechslera. 

20 µm 




Amphotericin B 0.25 Itraconazole 0.25 Voriconazole 0.06 


McGinnis and Pasarell (1998).

60 


Epicoccum purpurascens Ehrenb. ex Schlecht. 

Synonym: Epicoccum nigrum Link 

Colonies are fast growing, suede-like to downy, with a strong yellow to orange-brown 

diffusible pigment. When sporulating numerous black sporodochia (aggregates of 

conidiophores) are visible. Conidia are formed singly on densely compacted, nonspecialised, 

determinant, slightly pigmented conidiophores. Conidia are globose to 

pyriform, mostly l5-25 µm diameter with a funnel-shaped base and broad attachment 

scar, often seceding with a protuberant basal cell; i.e. aleuric or rhexolytic dehiscence 

of conidia. Conidia become multicellular (dictyoconidia), darkly pigmented and have a 

verrucose external surface. RG-1 organism. 

Epicoccum purpurascens is a cosmopolitan saprophyte of world-wide distribution which 

is occasionally isolated as a contaminant from clinical specimens like skin. 

Key Features: dematiaceous hyphomycete producing darkly pigmented, large globose 

to pyriform, verrucose dictyoconidia on a sporodochium. 


Domsch et al. (1980), McGinnis (1980) and Samson et al. (1995). 

15 µm 

Conidia of Epicoccum purpurascens.


Epidermophyton floccosum (Harz) Langeron et Milochevitch 

Colonies (SDA) are usually slow growing, greenish-brown or khaki-coloured with a 

suede-like surface, raised and folded in the centre, with a flat periphery and submerged 

fringe of growth. Older cultures may develop white pleomorphic tufts of mycelium. A 

deep yellowish-brown reverse pigment is usually present. Microscopic morphology 

shows characteristic smooth, thin-walled macroconidia which are often produced in 

clusters growing directly from the hyphae. Numerous chlamydospores are formed in 

older cultures. Microconidia are not formed. 

Epidermophyton floccosum is an anthropophilic dermatophyte with a world-wide distribution 

which often causes tinea pedis, tinea cruris, tinea corporis and onychomycosis. 

It is not known to invade hair in vivo and no specific growth requirements have been 

reported. RG-2 organism. 

Key Features: culture characteristics, microscopic morphology and clinical disease. 

For descriptions of species, keys to taxa and additional information see Rebell and 

Taplin (1970), Mackenzie et al. (1987), Rippon (1988) and de Hoog et al. (2000). 

20 µm 

15 µm 

Culture, macroconidia and chlamydospores of E. floccosum. 

Antifungal 

MIC µg/mL 


MIC µg/mL 

Range MIC90 Griseofulvin 0.06-2 1 Amphotericin B 0.03-0.5 0.25 

Itraconazole 0.01-8 0.125 Fluconazole 0.5->64 >64 

Terbinafine 0.01-1 0.06 Voriconazole 0.01-8 0.125 

Fernandez-Torres et al. (2001) and Sabatelli et al. (2006) and WCH in-house data.

62 


Exophiala dermatitidis (Kano) de Hoog 

Synonym: Wangiella dermatitidis (Kano) McGinnis 

Colonies are slow growing, initially yeast-like and black, becoming suede-like, olivaceous 

grey with the development of aerial mycelium with age. The initial yeast-like 

phase is referred to as the Phaeococcomyces exophialae synanamorph, which is characterised 

by unicellular, ovoid to elliptical, budding yeast-like cells. The yeast-like cells 

are hyaline and thin-walled when young becoming darkly pigmented (dematiaceous) 

and thick-walled when mature. With the development of mycelium, flask-shaped to cylindrical 

annellides are produced. Conidia are hyaline to pale brown, one-celled, round 

to obovoid, 2.0-4.0 x 2.5-6.0 µm, smooth-walled and accumulate in slimy balls at the 

apices of the annellides or down their sides. Cultures grow at 42 O C. 

E. dermatitidis has been isolated from plant debris and soil and is a recognised causative 

agent of mycetoma and phaeohyphomycosis in humans. RG-2 organism. 

For descriptions of species, keys to taxa and additional information see de Hoog and 

Hermanides-Nijhof (1977), McGinnis (1980), Hohl et al. (1983), Nishimura and Miyaji 

(1983), Matsumoto et al. (1984), Dixon and Polak-Wyss (1991), de Hoog et al. 

(2000). 

Antifungal 

10 µm 

Annellides and conidia of Exophiala dermatitidis. 

MIC µg/mL 

MIC µg/mL 


Range MIC90 Itraconazole 0.03-2 0.5 Amphotericin B 0.03-2 0.5 

Voriconazole 0.06-0.5 0.25 Posaconazole 0.03-1 nd 


McGinnis and Pasarell (1998), Espinel-Ingroff (2001), Espinel-Ingroff et al. (2001), 

Pfaller et al. (2002a) and WCH in-house data (nd = not done).


Exophiala jeanselmei Complex 

Synonyms: Phialophora jeanselmei (Langeron) Emmons 

E. jeanselmei has long been recognised as heterogeneous (de Hoog 1977). Recent 

molecular studies have redefined Exophiala jeanselmei and three additional species 

have been identified: E. oligosperma, E. nishimurae, and E. xenobiotica (Vitale and 

de Hoog, 2002 , de Hoog et al. 2003, 2006). These species are morphologically very 

similar and can best be distinguished by genetic analysis. 

Conidiogenous cells are predominantly annellidic and erect, multicellular conidiophores 

are absent. No growth at 40 O C. 

E. jeanselmei Mature conidiogenous cells rocket-shaped, slightly darker than 

the supporting hyphae, with regular tapering annellated zones. 

E. oligosperma Mature conidiogenous cells remain concolorous with supporting 

hyphae and may be intercalary and lateral, the latter being flask 

or rocket-shaped. Annellated zones have the appearance of inconspicuous 

flat scars. Chlamydospores are absent. 

E. nishimurae Similar morphology to E. oligosperma, however large chlamydospore-like 

cells are present. 

E. xenobiotica A segregant genotype of the E. jeanselmei complex with less 

melanised conidiogenous cells. 

Colonies are initially smooth, greenish-grey to black, mucoid and yeast-like, becoming 

raised and developing tufts of aerial mycelium with age, often becoming domeshaped 

and suede-like in texture. Reverse is olivaceous-black. Numerous ellipsoidal, 

yeast-like, budding cells are usually present, especially in young cultures. Scattered 

amongst these yeast-like cells are larger, inflated, subglobose to broadly ellipsoidal 

cells (germinating cells) which give rise to short torulose hyphae that gradually change 

into unswollen hyphae. Conidia are formed on lateral pegs either arising apically or laterally 

at right or acute angles from essentially undifferentiated hyphae or from strongly 

inflated detached conidia. Conidiogenous pegs are 1-3 µm long, slightly tapering and 

imperceptibly annellate. Conidia are hyaline, smooth, thin-walled, broadly ellipsoidal, 

3.2-4.4 x 1.2-2.2 µm, and with inconspicuous basal scars. Cultures grow at 37 O C but 

not at 40 O C. RG-2 organism. 

E. jeanselmei has a world-wide distribution and is a recognised causative agent of 

mycetoma and phaeohyphomycosis in humans. 


Hermanides-Nijhof (1977), de Hoog (1977, 1985), McGinnis and Padhye (1977), 

McGinnis (1978, 1980), Domsch et al. (1980), Nishimura and Miyaji (1983), Matsumoto 

et al. (1987), Dixon and Polak-Wyss (1991) and de Hoog et al. (2000, 2003, 2006). 

Antifungal 

MIC µg/mL 

MIC µg/mL 


Range MIC90 Itraconazole 0.03-2 0.5 Amphotericin B 0.03-4 0.5 

Voriconazole 0.06-2 0.5 Posaconazole 0.25-0.5 0.5 


McGinnis and Pasarell (1998), Espinel-Ingroff et al. (2001), Nucci et al. (2001) and 

WCH in-house data.

64 


Exophiala jeanselmei/spinifera Complex 

5 µm 

15 µm 

5 µm 

Annellides, conidia and conidiogenous pegs (annellides) on yeast-like cells and 

torulose hyphae of Exophiala jeanselmei. 

10 µm 

Erect, multiseptate conidiophores that are darker than the supporting hyphae, 

with long annellated zones and conidia of Exophiala spinifera.


Exophiala spinifera Complex 

Synonyms: Phialophora spinifera Nielsen & Conant 

Rhinocladiella spinifera (Nielsen & Conant) de Hoog 

Recent molecular studies have re-examined Exophiala spinifera and have recognised 

two species: E. spinifera and E. attenuata (Vitale and de Hoog, 2002). These two 

species are morphologically very similar and can best be distinguished by genetic 

analysis. 

Conidiogenous cells are predominately annellidic and erect, multicellular conidiophores 

that are darker than the supporting hyphae are present. No growth at 40 O C. 

E. spinifera Annellated zones are long with clearly visible, frilled annellations. 

E. attenuata Annellated zones are inconspicuous and degenerate. 

Colonies are initially mucoid and yeast-like, black, becoming raised and developing 

tufts of aerial mycelium with age, finally becoming suede-like to downy in texture. Reverse 

is olivaceous-black. Conidiophores are simple or branched, erect or sub-erect, 

spine-like with rather thick brown pigmented walls. Conidia are formed in basipetal succession 

on lateral pegs either arising apically or laterally at right or acute angles from 

the spine-like conidiophores or from undifferentiated hyphae. Conidiogenous pegs 

are 1-3 µm long, slightly tapering and imperceptibly annellate. Conidia are one-celled, 

subhyaline, smooth, thin-walled, subglobose to ellipsoidal, 1.0-2.9 x 1.8-2.5 µm, and 

aggregate in clusters at the tip of each annellide. Toruloid hyphae and yeast-like cells 

with secondary conidia are typically present. No growth at 40 O C. RG-2 organism. 

E. spinifera has a world-wide distribution and is a recognised causative agent of 

mycetoma and phaeohyphomycosis in humans. 


Hermanides-Nijhof (1977), McGinnis and Padhye (1977), Domsch et al. (1980), McGinnis 

(1980), Nishimura and Miyaji (1983), de Hoog (1985), Matsumoto et al. (1987), 

Dixon and Polak-Wyss (1991) and de Hoog et al. (2000, 2003, 2006). 

Antifungal 







66 


Exserohilum Leonard and Suggs 

Colonies are grey to blackish-brown, suede-like to floccose in texture and have an olivaceous 

black reverse. Conidia are straight, curved or slightly bent, ellipsoidal to fusiform 

and are formed apically through a pore (poroconidia) on a sympodially elongating 

geniculate conidiophore. Conidia have a strongly protruding, truncate hilum and the 

septum above the hilum is usually thickened and dark, with the end cells often paler 

than other cells, walls often finely roughed. Conidial germination is bipolar. 

The genus Exserohilum may be differentiated from the closely related genera Bipolaris 

and Dreschlera by forming conidia with a strongly protruding truncate hilum (i.e. 

exserted hilum). The hilum is defined as “a scar on a conidium at the point of attachment 

to the conidiophore”. In Drechslera species, the hilum does not protrude; in Bipolaris 

species the hilum protrudes only slightly. Several species of Exserohilum have 

been reported as agents of phaeohyphomycosis, notably E. rostratum (= E. halodes), 

E. mcginnisii and E. longirostratum. RG-1 organisms. 

Key Features: dematiaceous hyphomycete producing sympodial, transverse septate, 

ellipsoidal to fusiform conidia with a strongly protruding, truncate hilum. 

For descriptions of species, keys to taxa and additional information see Domsch et 

al. (1980), Alcorn (1983), Adam et al. (1986), McGinnis et al. (1986b), Rippon (1988), 

Burges et al. (1987), Dixon and Polak-Wyss (1991) and de Hoog et al. (2000). 

Antifungal 

20 µm 

20 µm 

Conidiophores and conidia with distinctive hilum (arrow) of E. rostratum. 








Fonsecaea pedrosoi/monophora Complex 

Morphologically the genus Fonsecaea is defined by the presence of indistinct melanised 

conidiophores with blunt, scattered denticles bearing conidia singly or in short 

chains that eventually become branched. de Hoog et al. (2004) revised the genus on 

the basis of ribosomal DNA internal transcribed spacer (ITS) sequence data recognising 

two species; F. pedrosoi and F. monophora. The previously described species F. 

compacta was found to be a morphological variant of F. pedrosoi. Morphological F. 

pedrosoi and F. monophora are very similar and can best be distinguished by genetic 

analysis. F. monophora on average has slightly longer conidial chains and slightly 

shorter denticles than F. pedrosoi. All strains grow at 37 O C but not at 40 O C. Both species 

are recognised etiologic agents of chromoblastomycosis. RG-2 organism. 

Colonies are slow growing, flat to heaped and folded, suede-like to downy, olivaceous 

to black with black reverse. Conidiogenous cells pale olivaceous, arranged in loosely 

branched systems, with prominent denticles. Conidia pale olivaceous, clavate to 

ellipsoidal, in short chains, subhyaline, smooth and thin-walled, 3.5-5 x 1.5-2 µm. 


(1980), Dixon and Polak-Wyss (1991), de Hoog et al. (2000), de Hoog et al. (2004). 

Antifungal 

Conidiophores and conidia of Fonsecaea. 

10 µm 

MIC µg/mL 

MIC µg/mL 


Range MIC90 Itraconazole 0.03-1 0.25 Amphotericin B 0.03-2 1 

Voriconazole 0.06-1 0.06 Posaconazole 0.06-1 nd 


McGinnis and Pasarell (1998), Espinel-Ingroff et al. (2001), Espinel-Ingroff (2003) 

and WCH in-house data (nd = not done).

68 


Fusarium Link ex Fries 

Colonies are usually fast growing, pale or bright coloured (depending on the species) 

with or without a cottony aerial mycelium. The colour of the thallus varies from whitish 

to yellow, pink, red or purple shades. Species of Fusarium typically produce both macro- 

and microconidia from slender phialides. Macroconidia are hyaline, two- to severalcelled, 

fusiform to sickle-shaped, mostly with an elongated apical cell and pedicellate 

basal cell. Microconidia are one- or two-celled, hyaline, smaller than macroconidia, 

pyriform, fusiform to ovoid, straight or curved. Chlamydospores may be present or 

absent. 

Cultures of F. oxysporum showing purple pigmentation 

and F. subglutinans showing pink pigmentation. 

Identification of Fusarium species is often difficult due to the variability between isolates 

(e.g. in shape and size of conidia and colony colour) and because not all features 

required are always well developed (e.g. the absence of macroconidia in some isolates 

after subculture). The important characters used in the identification of Fusarium species 

are as follows. Note: sporulation may need to be induced in some isolates and a 

good slide culture is essential. 

1. Colony growth diameters on potato dextrose agar and/or potato sucrose agar after 

incubation in the dark for 4 days at 25 O C. 

2. Culture pigmentation on potato dextrose agar and/or potato sucrose agar after incubation 

for 10-14 days with daily exposure to light. 

3. Microscopic morphology including shape of the macroconidia; presence or absence 

of microconidia; shape and mode of formation of microconidia; nature of the conidiogenous 

cell bearing microconidia; and presence or absence of chlamydospores. 

Most Fusarium species are soil fungi and have a world-wide distribution. Some are 

plant pathogens, causing root and stem rot, vascular wilt or fruit rot. Several species, 

notably F. oxysporum, F. solani and F. moniliforme are recognised as being pathogenic 

to man and animals causing hyalohyphomycosis (especially in burn victims and 

bone marrow transplant patients), mycotic keratitis and onychomycosis. Other species 

cause storage rot and are important mycotoxin producers. 

For descriptions of species, keys to taxa and additional information see Booth (1971 

and 1977), Domsch et al. (1980), McGinnis (1980), Burgess and Liddell (1983), Rippon 

(1988), Samson et al. (1995) and de Hoog et al. (2000).


15 µm 

Fusarium oxysporum Schlecht 

Colonies growing rapidly, 4.5 cm in 4 days, aerial mycelium white, becoming purple, 

with discrete orange sporodochia present in some strains; reverse hyaline to dark blue 

or dark purple. Conidiophores are short, single, lateral monophialides in the aerial 

mycelium, later arranged in densely branched clusters. Macroconidia are fusiform, 

slightly curved, pointed at the tip, mostly three septate, basal cells pedicellate, 23-54 x 

3-4.5 µm. Microconidia are abundant, never in chains, mostly non-septate, ellipsoidal 

to cylindrical, straight or often curved, 5-12 x 2.3 - 3.5 µm. Chlamydospores are terminal 

or intercalary, hyaline, smooth or rough-walled, 5-13 µm. In contrast to F. solani 

the phialides are short and mostly non-septate. RG-2 organism. 

Antifungal 

15 µm 

Microconidia on short phialides and macroconidia of F. oxysporum. 

MIC µg/mL 

MIC µg/mL 

Range MIC 

Antifungal 

Range MIC 90 90 

Itraconazole 0.5->16 >8 Amphotericin B 0.25->16 1-2 

Voriconazole 0.25->8 1-2 Posaconazole 1->8 4 


Espinel-Ingroff et al. (2001), Espinel-Ingroff (2001, 2003), Diekema et al. (2003), 

Cuenca-Estrella et al. (2006), Sabatelli et al. (2006) and WCH in-house data.

70 


Fusarium solani (Mart.) Sacc. 

Colonies growing rapidly, 4.5 cm in 4 days, aerial mycelium white to cream, becoming 

bluish-brown when sporodochia are present. Macroconidia are formed after 4-7 days 

from short multi-branched conidiophores which may form sporodochia. They are 3- to 

5- septate (usually 3- septate), fusiform, cylindrical, often moderately curved, with an 

indistinctly pedicellate foot cell and a short blunt apical cell, 28-42 x 4-6 µm. Microconidia 

are usually abundant, cylindrical to oval, one- to two-celled and formed from long 

lateral phialides, 8-16 x 2-4.5 µm. Chlamydospores are hyaline, globose, smooth to 

rough-walled, borne singly or in pairs on short lateral hyphal branches or intercalary, 

6-10 µm. RG-2 organism. 

Antifungal 

15 µm 

15 µm 

15 µm 

Microconidia on long phialides, macroconidia and chlamydospores of F. solani. 

MIC µg/mL 

MIC µg/mL 

Range MIC 

Antifungal 


Itraconazole 0.25->16 >8 Amphotericin B 0.25->16 4 

Voriconazole 0.125->8 4 (>8) Posaconazole >8 >8 


Espinel-Ingroff et al. (2001), Espinel-Ingroff (2001, 2003), Diekema et al. (2003), 

Cuenca-Estrella et al. (2006), Sabatelli et al. (2006) and WCH in-house data.


Geotrichum Link 

Species of the genus Geotrichum typically produce chains of hyaline, smooth, onecelled, 

subglobose to cylindrical arthroconidia by the holoarthric fragmentation of undifferentiated 

hyphae. Conidia may also develop sympodially and chlamydospores and 

endoconidia may also be present. The arthroconidia, which are quite variable in size 

may germinate at one end giving the appearance of a bud. However, the latter develops 

into a septate mycelium. True blastoconidia production is not found in the genus. 

This characteristic distinguishes the genus Geotrichum from Trichosporon which usually 

does produce blastoconidia. 

The need to exercise care when identifying species of Geotrichum must be stressed, 

as this name has often been used erroneously to describe any hyaline hyphomycete 

producing arthroconidia (McGinnis, 1980). Geotrichum species may be differentiated 

from each other using physiological and biochemical tests similar to those used for the 

identification of yeasts (Gueho, 1979 and Buchta and Otcenasek, 1988). 

For descriptions of species, keys to taxa and additional information see Gueho (1979), 

Domsch et al. (1980), McGinnis (1980), Barnett et al. (1983), Buchta and Otcenasek 

(1988), Samson et al. (1995), Kurtzman and Fell (1998) and de Hoog et al. (2000). 

Arthroconidium formation in G. candidum. Hyphal elements are progressively 

compartmentalised by fragmentation of septa. Conidial secession is 

by the centripetal separation (schizolysis) of a so called double septum and 

concomitant rupture of the original outer hyphal wall layer. 

Antifungal 

15 µm 

15 µm 

MIC µg/mL 

MIC µg/mL 

Range MIC90 Antifungal Range MIC90 Fluconazole 0.25-32 8-32 Amphotericin B 0.06-1.0 0.125 

Itraconazole 0.03->32 >32 Flucytosine 0.125-16 4 

Voriconazole 0.03-0.5 0.25 


Girmenia et al. (2003), Kucukates et al. (2005) and WCH in-house data.

72 


Geotrichum candidum Link 

Teleomorph: Galactomyces geotrichum (Butler & Petersen) Redhead & Malloch 

Colonies are fast growing, flat, white to cream, dry and finely suede-like with no reverse 

pigment. Hyphae are hyaline, septate, branched and break up into chains of hyaline, 

smooth, one-celled, subglobose to cylindrical arthroconidia. They are 6-12 x 3-6 µm in 

size and are released by the separation of a double septum. RG-1 organism. 

Physiological Tests: + Positive, - Negative, v Variable, w Weak, s Slow, n No data 

Germ Tube - L-Sorbose + L-Arabinose - D-Glucitol + 


Glucose v Maltose - D-Ribose v D-Gluconate - 


Sucrose - Trehalose + D-Glucosamine - myo-Inositol - 

Maltose - Lactose - N-A-D-glucosamine n 2-K-D-gluconate - 



Assimilation Melezitose - Ribitol v Urease - 


Galactose + D-Xylose + D-Mannitol v Growth at 37 O C v 

Geotrichum candidum is an extremely common fungus with a world-wide distribution. 

Pulmonary involvement is the most frequently reported form of the disease, but bronchial, 

oral, vaginal, cutaneous and alimentary infections have also been noted. 

Geotrichum capitatum (Diddens & Lodder) v. Arx 

Teleomorph: Dipodascus capitatis de Hoog et al. 

Synonyms: Trichosporon capitatum, Blastoschizomyces capitis 

Colonies are moderately fast growing, flat, whitish, and finely suede-like with no reverse 

pigment. Hyphae are profusely branched at acute angles, with terminal and intercalary 

conidiogenous cells which form long, cicatrized rachids on which conidia are borne. 

Conidia are hyaline, smooth, one-celled, cylindrical to clavate, with a rounded apex 

and flat base, 7-10 x 2.5-3.5 µm. Rectangular arthroconidia are also often present. 


Physiological Tests: + Positive, - Negative, v Variable, w Weak, s Slow, n No data 

Germ Tube - L-Sorbose v L-Arabinose - D-Glucitol - 


Glucose - Maltose - D-Ribose v D-Gluconate - 



Maltose - Lactose - N-A-D-glucosamine n 2-K-D-gluconate - 





Galactose + D-Xylose - D-Mannitol - Growth at 40 O C + 

Geotrichum capitatum occurs quite commonly in humans, usually as a transient component 

of normal skin flora and sputum. Systemic infections including pulmonary, 

fungemia and endocarditis have been reported in immunosuppressed patients.


Gliocladium Corda 

The genus Gliocladium is often described as a counterpart of Penicillium with slimy 

conidia. Colonies are fast growing, suede-like to downy in texture, white at first, sometimes 

pink to salmon, becoming pale to dark green with sporulation. The most characteristic 

feature of the genus is the distinctive erect, often densely penicillate conidiophores 

with phialides which bear slimy, one-celled hyaline to green, smooth-walled 

conidia in heads or columns. Although, some penicillate conidiophores are always 

present, Gliocladium species may also produce verticillate branching conidiophores 

which can be confused with Verticillium or Trichoderma. 

Gliocladium species have a world-wide distribution and are commonly isolated from a 

wide range of plant debris and soil. RG-1 organism. 

Key Features: hyphomycete producing distinctive erect penicillate conidiophores with 

phialides bearing clusters of single-celled conidia. 


(1980), McGinnis (1980), Onions et al. (1981), Rippon (1988), de Hoog et al. (2000). 

Conidiophore and conidia of Gliocladium. 

10 µm

74 


Graphium Corda 

The genus Graphium is characterised by the formation of synnemata which consist of 

a more or less compact group of erect conidiophores that are cemented together, usually 

splaying out and bearing conidia at the apex. Synnemata are darkly pigmented, 

erect and occur solitary or in clusters. Conidia are hyaline, one-celled, smooth, subglobose 

to ovoid and are usually aggregated in slimy heads at the apex of the synnemata. 

Colonies are effuse, grey, olivaceous brown or black. 

Graphium eumorphum is one of the synanamorphs of Pseudallescheria boydii and is 

commonly found on woody plant material. RG-1 organism. 

Key Features: dematiaceous hyphomycete producing erect synnemata with apical 

aggregates of single-celled conidia in slimy heads. 

For descriptions of species, keys to taxa and additional information see Barron (1968), 

Ellis (1971), McGinnis (1980) and de Hoog et al. (2000). 

20 µm 

Synnemata and conidia of Graphium.


Histoplasma capsulatum Darling 

Histoplasma capsulatum exhibits thermal dimorphism growing in living tissue or in culture 

at 37 O C as a budding yeast-like fungus and in soil or culture at temperatures below 

30 O C as a mould. 

Colonies (SDA) at 25 O C are slow growing, white or buff-brown, suede-like to cottony 

with a pale yellow-brown reverse. Other colony types are glabrous or verrucose, and a 

red pigmented strain has been noted (Rippon, 1988). Microscopic morphology shows 

the presence of characteristic large, rounded, single-celled, 8-14 µm in diameter, tuberculate 

macroconidia formed on short, hyaline, undifferentiated conidiophores. Small, 

round to pyriform, 2-4 µm in diameter, microconidia borne on short branches or directly 

on the sides of the hyphae may also be present. 

On brain heart infusion agar containing blood incubated at 37 O C colonies are smooth, 

moist, white and yeast-like. Microscopically, numerous small round to oval budding 

yeast-like cells, 3-4 x 2-3 µm in size are observed. 

Three varieties of Histoplasma capsulatum are recognised, depending on the clinical 

disease: var. capsulatum is the common cause of histoplasmosis; var. duboisii is the 

African type and var. farciminosum causes lymphangitis in horses. Histoplasma isolates 

may also resemble species of Sepedonium and Chrysosporium. Traditionally, 

positive identification required conversion of the mould form to the yeast phase by 

growth at 37 O C on enriched media, however culture identification by exoantigen test is 

now the method of choice. 

WARNING: RG-3 organism. Cultures of Histoplasma capsulatum represent a severe 

biohazard to laboratory personnel and must be handled with extreme caution in an 

appropriate pathogen handling cabinet. 

Key Features: clinical history, tissue morphology, culture morphology and positive 


Histoplasma capsulatum has a world wide distribution, however the Mississippi-Ohio 

River Valley in the USA is recognised as a major endemic region. Environmental isolations 

of the fungus have been made from soil enriched with excreta from chicken, starlings 

and bats. Histoplasmosis is an intracellular mycotic infection of the reticuloendothelial 

system caused by the inhalation of the fungus. Approximately 95% of cases 

of histoplasmosis are inapparent, subclinical or benign. Five percent of the cases have 

chronic progressive lung disease, chronic cutaneous or systemic disease or an acute 

fulminating fatal systemic disease. All stages of this disease may mimic tuberculosis. 

Sporadic cases do occur in Australia. 


(1980), Chandler et al. (1980), George and Penn (1986), Rippon (1988) and de Hoog 

et al. (2000).

76 


20 µm 

Culture and microscopic morphology of the saprophytic or mycelial form of 

H. capsulatum showing characteristic large, rounded, single-celled, tuberculate 

macroconidia formed on short, hyaline, undifferentiated conidiophores. 

Antifungal 

MIC µg/mL 


MIC µg/mL 

Range MIC90 Amphotericin B 0.03-2 0.5-1 Posaconazole 0.25-1 0.25 (2) 

Fluconazole 0.125-64 16 Voriconazole 0.03-2 0.25 (1) 

Itraconazole 0.03-8 0.06 (1) 

Histoplasma capsulatum Darling 

Limited data available. Espinel-Ingroff et al. (2001), Espinel-Ingroff (2003), Gonzales 

et al. (2005) and Sabatelli et al. (2006).


Synonyms: Phaeoannellomyces werneckii (Horta) McGinnis & Schell 

Exophiala werneckii (Horta) v. Arx 

Colonies are slow growing, initially mucoid, yeast-like and shiny black. However with 

age they develop abundant aerial mycelia and become dark olivaceous in colour. Microscopically, 

colonies consist of brown to dark olivaceous, septate hyphal elements 

and numerous two-celled, pale brown, cylindrical to spindle-shaped yeast-like cells that 

taper towards the ends to form an annellide. Most yeast-like cells also have prominent 

darkly-pigmented septa. Annellides may also arise from the hyphae. Conidia are one 

to two-celled, cylindrical to spindle-shaped, hyaline to pale brown and usually occur in 

aggregated masses. RG-1 organism. 

Hortaea werneckii is a common saprophytic fungus believed to occur in soil, compost, 

humus and on wood in humid tropical and sub-tropical regions and is the causative 

agent of tinea nigra in humans. 

Key Features: dematiaceous hyphomycete, two-celled yeast-like cells producing 

annelloconidia. 

For description of species, keys to taxa and additional information see Mok (1982), 

McGinnis (1980), McGinnis et al. (1985), Rippon (1988) and de Hoog et al. (2000). 

Antifungal 

Hortaea werneckii (Horta) Nishimura & Miyaji 

Culture and conidia of Hortaea werneckii. 

20 µm 







78 


Lasiodiplodia theobromae (Pat.) Griffon & Maublanc 

Teleomorph: Botryosphaeria rhodina (Berk, & Curt. v. Arx 

Synonym: Botryodiplodia theobromae Patouillard 

Colonies are greyish sepia to mouse grey to black, fluffy with abundant aerial mycelium; 

reverse fuscous black to black. Pycnidia are simple or compound, often aggregated, 

stromatic, ostiolate, frequently setose, up to 5 mm wide. Conidiophores are hyaline, 

simple, sometimes septate, rarely branched cylindrical, arising from the inner layers of 

cells lining the pycnidial cavity. Conidiogenous cells are hyaline, simple, cylindrical to 

subobpyriform, holoblastic, annellidic. Conidia are initially unicellular, hyaline, granulose, 

subovoid to ellipsoide-oblong, thick-walled, base truncate; mature conidia oneseptate, 

cinnamon to fawn, often longitudinally striate, 20-30 x 10-15 µm. Paraphyses 

when present are hyaline, cylindrical, sometimes septate, up to 50 µm long. 

Lasiodiplodia theobromae is a well known plant pathogen and it has been reported 

from about 500 host plants, mainly confined to an area 40 O north to 40 O south of the 

equator. It has also been associated with mycotic keratitis, lesions on nail and subcutaneous 

tissue. RG-1 organism. 

Key Features: coelomycete, with pycnidia producing characteristic two-celled, dark 

brown, striated conidia. 

For description of species, keys to taxa and additional information see de Hoog et al. 

(2000). 

10 µm 

Mature two-celled dark brown conidia with typical striations of L. theobromae. 

Antifungal 




Amphotericin B 0.03 Itraconazole 16 Voriconazole 0.25 


WCH in-house data only.


Lecythophora Nannfeldt 

Colonies are pink to salmon, later becoming blackish, smooth, often mucoid or yeastlike. 

Conidiogenous cells are poorly differentiated, usually lateral or intercalary, hyphae 

bearing one or several scattered inconspicuous collarettes, either sessile or on small 

outgrowths. Conidia are hyaline, smooth and thin walled, broadly ellipsoidal, cylindrical, 

reniform or allantoid. Chlamydospores may be present. Lecythophora contains 6 

species, with two species of medical interest; L hoffmannii and L. mutabilis. 

Lecythophora hoffmannii (J.F.H. Beyma) W. Gams & McGinnis 

Teleomorph: Coniochaeta ligniaria (Grev.) Cooke 

Colonies are flat, smooth, moist, pink to orange, with regular and sharp margin; reverse 

pink. Hyphae are narrow, hyaline, producing conidia laterally from small collarettes 

directly on the hyphae, or from lateral cells which are sometimes arranged in dense 

groups; lateral cells flask-shaped or nearly cylindrical. Collarettes are unpigmented, 

about 1.5 µm wide. Conidia are hyaline, smooth and thin walled, broadly ellipsoidal to 

cylindrical or allantoid, 3.0-3.5 x 1.5-2.5 µm, produced in slimy heads. RG-1 organism. 

For description of species, keys to taxa and additional information see de Hoog (1983) 


10 µm 

10 µm 

Culture, hyphae with small collarettes and conidia of Lecythophora hoffmannii. 

Antifungal 




Amphotericin B 0.06-0.5 Itraconazole 0.06-32 Voriconazole 0.125-0.5 


McGinnis and Pasarell (1998).

80 


Madurella grisea Mackinnon, Ferrada and Montemayer 

The genus Madurella is based on tissue morphology (mycetoma with black grains) 

and the formation of sterile cultures on mycological media. Both M. mycetomatis and 

M. grisea have been isolated from soil and are one of the major causative agents of 

mycetoma. 

Colonies are slow growing, dark, leathery, folded with radial grooves and with a light 

brown to greyish surface mycelium. With age, colonies become dark brown to reddish-brown 

and have a brownish-black reverse. Microscopically, cultures are sterile, 

although hyphae of two widths have been described, thin at 1-3 µm in width or broad at 

3-5 µm in width. The optimum temperature for growth of M. grisea is 30 O C; this fungus 

does not grow at 37 O C. RG-2 organism. 

M. grisea can be distinguished from Madurella mycetomatis by the inability to grow at 

37 O C and to assimilate lactose. 

Key Features: black grain mycetoma, no growth at 37 O C, no diffusible brown pigment 

produced on culture and absence of conidia. 


(1980), Chandler et al. (1980), Rippon (1988) and de Hoog et al. (2000). 

Antifungal 

100 µm 

Grains of Madurella grisea (tissue microcolonies) are black, round to 

lobed, soft to firm, up to 1.0 mm, with two distinctive zones, a hyaline to 

weakly pigmented central zone and a deeply pigmented periphery. 




Amphotericin B 0.25 Itraconazole 0.5 Voriconazole 0.5 


Espinel-Ingroff et al. (2001).


Madurella mycetomatis (Laveran) Brumpt 

Colonies are slow growing, flat and leathery at first, white to yellow to yellowish-brown, 

becoming brownish, folded and heaped with age and with the formation of aerial mycelia. 

A brown diffusible pigment is characteristically produced in primary cultures. Although 

most cultures are sterile, two types of conidiation have been observed, the first 

being flask-shaped phialides that bear rounded conidia, the second being simple or 

branched conidiophores bearing pyriform conidia (3-5 µm) with truncated bases. The 

optimum temperature for growth of this mould is 37 O C. RG-2 organism 

Grains of Madurella mycetomatis (tissue microcolonies) are brown or black, 0.5-1.0 

mm in size, round or lobed, hard and brittle, composed of hyphae which are 2-5 µm in 

diameter, with terminal cells expanded to 12-15 (30) µm in diameter. 

M. mycetomatis can be distinguished from Madurella grisea by growth at 37 O C and its 

inability to assimilate sucrose. 

Key Features: black grain mycetoma, growth at 37 O C, diffusible brown pigment 

produced on culture and the occasional presence of phialides. 



20 µm 

Culture showing brown diffusible pigment and phialides of M. mycetomatis. 

Antifungal 




Amphotericin B 0.03 Itraconazole 0.03-0.125 Voriconazole 0.03-0.6 


McGinnis and Pasarell (1998) and Espinel-Ingroff et al. (2001).

82 


Malassezia Baillon 

Malassezia is characterised by globose, oblong-ellipsoidal to cylindrical, yeast cells. 

Reproduction is by budding on a broad base and from the same site at one pole 

(unipolar). With the exception of M. pachydermatis, Malassezia are lipophilic yeasts, 

therefore in vitro growth must be stimulated by natural oils or other fatty substances. 

The most common method used is to overlay Sabouraud’s dextrose agar containing 

cycloheximide (actidione) with olive oil or alternatively to use a more specialised media 

like Dixon’s agar which contains glycerol mono-oleate. On such media, colonies are 

cream to yellowish, smooth or lightly wrinkled, glistening or dull, and with the margin 

being either entire or lobate (see photo). Seven species have now been recognised 

(Gueho et al. 1996). RG-2 organisms. 

Species Source 

M. furfur humans, normal flora, pityriasis 

M. globosa humans, normal flora, pityriasis 

M. pachydermatis animals, especially dogs 

M. obtusa humans, normal flora, 

atopic dermatitis 

M. restricta humans, normal flora 

M. slooffiae human and pig normal flora 

M. sympodialis humans, normal flora 

Identification criteria for the differentiation of Malassezia species (de Hoog et. al. 2000). 

Buds SDA 40 O C Cremophor 

EL 

Tween 

80 

Tween 

40 

Tween 

20 

Esculine Catalase 

M. furfur wide - + + + + + w + 

M. globosa narrow - - - - - - - + 

M. pachydermatis wide + + -,w + + -,w v v 

M. obtusa wide - - - - - - + + 

M. restricta narrow - - - - - - - - 

M. slooffiae wide - + - +,w + + - + 

M. sympodialis narrow - + -,w + + + + + 

For descriptions of species, keys to taxa and additional information see Guillot and 

Gueho (1995), Gueho et al. (1996), Guillot et al. (1996, 2000), de Hoog et al. (2000). 

Antifungal 

MIC µg/mL 

MIC µg/mL 

Range MIC90 Antifungal Range MIC90 Fluconazole 0.125->64 4 (8) Amphotericin B 0.03-16 1 (8) 

Itraconazole 0.03-16 0.125 Voriconazole 0.03-16 0.125 (1) 

Ketoconazole 0.03-4 0.25 Posaconazole 0.03-32 0.125 (2) 

Very limited data available. Special growth conditions are needed for antifungal 

susceptibility testing. Nakamura et al. (2000), Velegraki et al. (2004) and Miranda et 

al. (2007).


Malbranchea Saccardo 

Colonies are white to sulphur-yellow to ochre-brown in colour, suede-like in texture, 

with a reddish-brown reverse, and often a reddish diffusible pigment. Microscopic morphology 

shows typical hyaline, one-celled, cylindrical, truncate, alternate arthroconidia 

produced in terminal fertile portions of the hyphae. Arthroconidia are released by lysis 

of the disjunctor cells. These arthroconidia may be perceived as a yellow dust when 

released at maturity. RG-1 organisms. 

Key Features: hyphomycete producing alternate arthroconidia with disjunctor cells. 

Malbranchea species are soil fungi of world-wide distribution which microscopically 

may resemble Coccidioides immitis/posadasii. Exoantigen tests are now the method 

of choice for culture identification of C. immitis/posadasii. 

For description of the species, keys to taxa and additional information see Cooney and 

Emerson (1964), McGinnis (1980), Rippon (1988) and de Hoog et al. (2000). 

20 µm 

Arthroconidia of Malbranchea.

84 


Microsporum Gruby 

Teleomorph: Arthroderma Currey and Berkeley emend Weitzman et al. 

Microsporum species form both macro- and microconidia on short conidiophores. 

Macroconidia are hyaline, multiseptate, variable in form, fusiform, spindle-shaped to 

obovate, ranging from 7-20 x 30-60 µm in size, with thin- or thick- echinulate to verrucose 

cell walls. Their shape, size and cell wall features are important characteristics 

for species identification. Microconidia are hyaline, single-celled, pyriform to clavate, 

smooth-walled, 2.5-3.5 x 4-7 µm in size and are not diagnostic for any one species. 

The separation of this genus from Trichophyton is essentially based on the roughness 

of the macroconidial cell wall, although in practice this may sometimes be difficult to 

observe. Seventeen species of Microsporum have been described (Rippon, 1988) 

however only the more common species are included in these descriptions. 

It is essential to observe macroconidia when identifying species of Microsporum. 

Strains of M. canis often do not produce macroconidia and/or microconidia on primary 

isolation media and it is recommended that sub-cultures be made onto polished rice 

grains to stimulate sporulation. These non-sporulating strains of M. canis are often 

erroneously identified as M. audouinii and it is surprising just how many laboratories 

have difficulty in differentiating between M. canis and M. audouinii. 

For descriptions of species, keys to taxa and additional information see Rebell and 

Taplin (1970), Vanbreusegham et al. (1978), Rippon (1988), McGinnis (1980), Domsch 

et al. (1980), Ajello (1977), Weitzman et al. (1986), Mackenzie et al. (1986), Kane et al. 

(1997) and de Hoog et al. (2000). 

a 

(a) M. audouinii showing poor growth on rice grains, usually being visible only 

as a brown discolouration. (b) M. canis on rice grains showing good growth, 

yellow pigmentation and sporulation. 

Antifungal 

MIC µg/mL 

MIC µg/mL 

Range MIC90 Antifungal Range MIC90 Griseofulvin 0.125-2 1 Amphotericin B 0.03-8 1-2 

Itraconazole 0.01-4 0.25-0.5 Fluconazole 0.06->64 64 


Fernandez-Torres et al. (2001), Sabatelli et al. (2006) and WCH in-house data. 

b


Microsporum audouinii Gruby 

Colonies (SDA) are flat, spreading, greyish-white to light tan-white in colour, and have 

a dense suede-like to downy surface, suggestive of mouse fur in texture. Reverse 

can be yellow-brown to reddish-brown in colour. Some strains may show no reverse 

pigment. Macroconidia and microconidia are only rarely produced, most cultures are 

sterile or produce only occasional thick-walled terminal or intercalary chlamydospores. 

When present macroconidia may resemble those of M. canis but are usually longer, 

smoother and more irregularly fusiform in shape; microconidia when present are pyriform 

to clavate in shape and are similar to those seen in other species of Microsporum. 

So called pectinate (comb-like) hyphae and racquet hyphae (a series of hyphal segments 

swollen at one end) may also be present. RG-2 organism. 

Culture and a thick-walled intercalary chlamydospore of M. audouinii. Note: 

macroconidia and microconidia are only rarely produced. 

Growth on Rice Grains: Very poor or absent, usually being visible only as a brown 

discolouration. This is one of the features which distinguish M. audouinii from M. 

canis. 

Reverse Pigment on Potato Dextrose Agar: Salmon to pinkish-brown (M. canis is 

bright yellow). 

Lactritmel Agar: Colonies are usually flat, spreading, with a fine, whitish suede-like 

surface and a very pale yellow-brown reverse. Microscopic morphology as described 

above. 

Vitamin Free Agar (Trichophyton Agar No.1): Good growth indicating no special 

nutritional requirements. Cultures are flat, white, suede-like to downy, with a yellowbrown 

reverse. Note: growth of some strains of M. audouinii is enhanced by the presence 

of thiamine (Trichophyton agar No.4). 

Hair Perforation Test: Negative after 28 days. 

10 µm 

Key Features: absence of conidia, poor or no growth on polished rice grains, inability 

to perforate hair in vitro, and culture characteristics. 

M. audouinii is an anthropophilic fungus causing non-inflammatory infections of scalp 

and skin especially in children. Once the cause of epidemics of tinea capitis in Europe 

and North America, it is now becoming less frequent. Invaded hairs show an ectothrix 

infection and usually fluoresce a bright greenish-yellow under Wood’s ultra-violet light. 

Only rarely found in Australasia, most reports are in fact non-sporulating strains of M. 

canis.

86 


Microsporum canis Bodin 

Teleomorph: Arthroderma otae (Hasegawa and Usui) McGinnis, et al. 

Colonies (SDA) are flat, spreading, white to cream-coloured, with a dense cottony 

surface which may show some radial grooves. Colonies usually have a bright golden 

yellow to brownish yellow reverse pigment, but non-pigmented strains may also occur. 

Macroconidia are typically spindle-shaped with 5-15 cells, verrucose, thick-walled and 

often have a terminal knob, 35-110 x 12-25 µm. A few pyriform to clavate microconidia 

are also present. Macroconidia and/or microconidia are often not produced on primary 

isolation media and it is recommended that sub-cultures be made onto Lactritmel Agar 

and/or boiled polished rice grains to stimulate sporulation. RG-2 organism. 

Growth on Rice Grains: good growth of white aerial mycelium with production of yellow 

pigment. Microscopy reveals numerous macroconidia and microconidia similar to 

those described above. 

Lactritmel Agar: Flat, white suede-like to cottony colony with a bright yellow reverse. 

Microscopy reveals moderate numbers of thick-walled, multiseptate, long, spindleshaped 

macroconidia, some of which show a terminal knob. Walls of macroconidia 

are slightly rough or echinulate especially at terminal knobs. Numerous clavate to 

pyriform microconidia are also present in this strain. Lactritmel agar with sterile soil 

added is also an excellent medium for the stimulation of macroconidial development 

in M. canis. 

Reverse Pigment on Potato Dextrose Agar: Bright yellow (both M. audouinii and M. 

canis var. equinum are salmon to pinkish-brown). 


nutritional requirements. Cultures are flat, white, suede-like to downy, with a yellow to 

pale yellow-brown reverse. 

Hair Perforation Test: Positive at 14 days. 

Key Features: distinctive macroconidia and culture characteristics. Abundant growth 

and sporulation on polished rice grains and in vitro perforation of hair. 

Culture of Microsporum canis.


Microsporum canis Bodin 

Typical spindle-shaped macroconidia of Microsporum canis. 

20 µm 

Dysgonic strains of M. canis are rare but may also occur. These dysgonic strains have 

a typically heaped and folded, yellow-brown thallus and macroconidia are usually 

absent. However, typical colonies and macroconidia of M. canis are usually produced 

by this variant when subcultured onto polished rice grains. Note: the dysgonic type 

colony of M. canis is similar to that of Microsporum ferrugineum. 

M. canis is a zoophilic dermatophyte of world-wide distribution and is a frequent cause 

of ringworm in humans, especially children. Invades hair, skin and rarely nails. Cats 

and dogs are the main sources of infection. Invaded hairs show an ectothrix infection 

and fluoresce a bright greenish-yellow under Wood’s ultra-violet light.

88 


Microsporum canis var. distortum di Menna & Marples 

Supplementary description for Microsporum canis var. distortum, a dysgonic variant of 

M. canis with distinctive distorted macroconidia. Abundant growth and sporulation on 

rice grains. RG-2 organism. 

Microsporum canis var. distortum is a zoophilic fungus known to cause infections in 

cats, dogs and other animals. It is a rare cause of tinea capitis in New Zealand, 

Australia and North America. Clinical disease is similar to M. canis . Invaded hairs 

show an ectothrix infection and fluoresce a bright greenish-yellow under Wood’s ultraviolet 

light. 

20 µm 

Culture and distorted macroconidia of M. canis var. distortum.


Microsporum canis var. equinum (Delacroix & Bodin) comb. nov. 

Basionym: Microsporum equinum (Delacroix & Bodin) Gueguen 

Microsporum equinum is now considered to be a genotypic synonym of Microsporum 

canis (de Hoog et al. 2000), however we propose it be maintained as a variety of M. 

canis due to phenotypic and epidemiologic differences. 

Colonies are flat, spreading, suede-like, pale buff to pale salmon, usually with some radial 

furrows. A buff to pinkish-buff to yellow-brown reverse pigment is produced. Note: 

only a few strains produce conidia on primary isolation, however growth on urea agar 

usually stimulates the production of macroconidia. Macroconidia are small (especially 

when compared to those produced by M. canis), broad, irregular, spindle-shaped, 18- 

60 x 5-15 mm with rough thick walls and few septa. Microconidia are pyriform to clavate 

in shape, 3-9 x 1.5-3.5 mm, but are rarely produced. RG-2 organism. 

Growth on Rice Grains: Poor with minimal surface growth, usually being visible only 

as a brown discolouration. This is one of the features which distinguish M. canis var. 

equinum from M. canis. 

Reverse Pigment on Potato Dextrose Agar: Salmon to pinkish-brown (M. canis is 

bright yellow). 

Lactritmel Agar: Colonies are usually flat, spreading, with a fine, whitish suede-like 

surface and a very pale yellow-brown reverse. Microscopic morphology as described 

above. 


nutritional requirements. Cultures are flat, white, suede-like to downy, with a yellowbrown 

reverse. 

Hair Perforation Test: Negative after 28 days. 

Key Features: macroconidial shape and size, inability to perforate hair in vitro, and 

poor growth on polished rice grains. 

20 µm 

Culture and macroconidia of M. canis var. equinum. 

Microsporum equinum is a 

rare cause of ringworm of 

horses. Invaded hairs show 

an ectothrix infection and 

fluoresce a bright greenishyellow 

under Wood’s ultraviolet 

light. Rarely infects 

man or other animal species. 

Reported from Australia, 

Europe and North America.

90 


Teleomorph: Arthroderma cajetani Ajello, Weitzman, McGinnis & Padhye 

Colonies (SDA) are flat, spreading, buff to pale brown, powdery to suede-like, with a 

slightly raised and folded centre and some radial grooves. Reverse pigment dark reddish 

brown. Numerous large, very thick-walled, echinulate (rough) elliptical macroconidia 

with predominantly 5-6 septa but may be 2-8 septate. Occasional spirals may be 

seen. Moderate numbers of mainly slender clavate with some pyriform microconidia 

are present. The macroconidia are quite characteristic and diagnostic of M. cookei and 

further tests are not necessary. The thick walls and usually larger size of the macroconidia 

distinguish M. cookei from M. gypseum. RG-1 organism. 

Lactritmel Agar: Flat, buff-coloured, suede-like to powdery colony with a deep redbrown 

reverse. Microscopic morphology as described above for the primary culture. 


nutritional requirements, pinkish-buff-coloured, suede-like colony with a deep magenta 

red reverse. 

Hair Perforation Test: Positive. 

Microsporum cookei Ajello 

Key Features: distinctive macroconidial morphology and culture characteristics. 

20 µm 

Culture and macroconidia of Microsporum cookei. 

Microsporum cookei is a geophilic fungus which has been isolated from hair of small 

mammals showing no clinical lesions. Infection has been reported in rodents, dogs 

and rarely in humans. It is not known to invade hair in vivo, but produces hair perforations 

in vitro. M. cookei has a world-wide distribution.


Microsporum ferrugineum Ota 

Colonies (SDA) are slow growing, forming a waxy, glabrous, convoluted thallus with a 

cream to buff-coloured surface and no reverse pigment. Note: surface pigmentation 

may vary from cream to yellow to deep red and a flatter white form sometimes occurs. 

Cultures rapidly become downy and pleomorphic. Microconidia or macroconidia are 

not produced. However, irregular branching hyphae with prominent cross walls (“bamboo 

hyphae”) and chlamydospores are seen. The so-called “bamboo hyphae” is a 

characteristic of this species. RG-2 organism. 

Key Features: clinical history, culture characteristics and distinctive “bamboo” 

hyphae. 

20 µm 

Culture and “bamboo hyphae” of Microsporum ferrugineum. 

Microsporum ferrugineum is an anthropophilic fungus causing epidemic juvenile tinea 

capitis in humans. The clinical features are similar to those of infections caused by 

M. audouinii. Invaded hairs show an ectothrix infection and fluoresce a greenish-yellow 

under Wood’s ultra-violet light. Reported from Asia (including China and Japan), 

USSR, Eastern Europe and Africa.

92 


Microsporum fulvum Uriburu 

Teleomorph: Arthroderma fulvum (Stockdale) Weitzman et al. 

Colonies (SDA) are fast growing, flat, suede-like, tawny-buff to pinkish-buff in colour and 

frequently have a fluffy white advancing edge. A dark red under surface is occasionally 

seen, otherwise it is colourless to yellow brown. Abundant thin-walled, elongate, ellipsoidal 

macroconidia are formed which closely resemble those of M. gypseum, except 

they are longer and more bullet-shaped (clavate) with 3 to 6 septa. Numerous spiral 

hyphae, which are often branched are seen. Numerous pyriform to clavate microconidia 

are also produced but these are not diagnostic. RG-1 organism. 

Key Features: macroconidial morphology and culture characteristics. 

20 µm 

Culture and macroconidia of Microsporum fulvum. 

Microsporum fulvum is a geophilic fungus of world-wide distribution which may cause 

occasional infections in humans and animals. Clinical disease is similar to M. gypseum 

but less common. Invaded hairs show a sparse ectothrix infection but do not fluoresce 

under Wood’s ultra-violet light.


Microsporum gallinae (Megnin) Grigorakis 

Colonies (SDA) are flat with a suede-like texture and are white in colour with a pinkish 

tinge. Some cultures show radial folding. An orange-pink “strawberry-coloured” 

reverse pigment is usually present. Macroconidia when present are usually five- to sixcelled, 

thin to thick-walled, slightly echinulate, cylindrical to clavate with narrow base 

and blunt tip, 15-60 x 6-10 µm. Microconidia are ovoidal to pyriform in shape. RG-2 

organism. 

Key Features: macroconidial morphology, culture characteristics and clinical lesions 

in chickens. 

Culture and macroconidia of Microsporum gallinae. 

20 µm 

Microsporum gallinae is a zoophilic fungus causing fowl favus in chickens and other 

fowl, affecting the comb and wattles producing “white comb” lesions. A rare cause of 

tinea in humans. Invaded hairs show a sparse ectothrix infection but do not fluoresce 

under Wood’s ultra-violet light.

94 


Microsporum gypseum (Bodin) Guiart & Grigorakis 

Teleomorphs: Arthroderma gypsea (Nannizzi) Weitzman et al. 

Arthroderma incurvatum (Stockdale) Weitzman et al. 

Colonies (SDA) are usually flat, spreading, suede-like to granular, with a deep cream 

to tawny-buff to pale cinnamon-coloured surface. Many cultures develop a central 

white downy umbo (dome) or a fluffy white tuft of mycelium and some also have a narrow 

white peripheral border. A yellow-brown pigment, often with a central darker brown 

spot, is usually produced on the reverse, however a reddish-brown reverse pigment 

may be present in some strains. Cultures produce abundant, symmetrical, ellipsoidal, 

thin-walled, verrucose, four- to six-celled macroconidia. The terminal or distal ends of 

most macroconidia are slightly rounded, while the proximal ends (point of attachment 

to hyphae) are truncate. Numerous clavate-shaped microconidia are also present, but 

these are not diagnostic. RG-1 organism. 

Key Features: distinctive macroconidia and culture characteristics. 

20 µm 

Culture and macroconidia of Microsporum gypseum. 

Microsporum gypseum is a geophilic fungus with a world-wide distribution which may 

cause infections in animals and humans, particularly children and rural workers during 

warm humid weather. Usually produces a single inflammatory skin or scalp lesion. Invaded 

hairs show an ectothrix infection but do not fluoresce under Wood’s ultra-violet 

light.


Microsporum nanum Fuentes 

Teleomorph: Arthroderma obtusum (Dawson and Gentles) Weitzman et al. 

Colonies (SDA) are flat, cream to buff in colour with a suede-like to powdery surface 

texture. Young colonies have a brownish-orange pigment which deepens into a dark 

reddish-brown with age. Cultures produce numerous small ovoid to pyriform macroconidia 

with one to three (mostly 2) cells, with relatively thin, finely echinulate (rough) 

walls, and broad truncate bases. Many macroconidia are borne on conidiophores 

(stalks) which do not stain readily. Occasional clavate microconidia are present, which 

distinguishes M. nanum from some species of Chrysosporium. RG-2 organism. 

Key Features: distinctive macroconidia and culture characteristics. 

20 µm 

Culture and macroconidia of Microsporum nanum. 

Microsporum nanum is a geophilic and zoophilic fungus frequently causing chronic 

non-inflammatory lesions in pigs and a rare cause of tinea in humans. Also present in 

soil of pig-yards. Infections in man are usually contacted directly from pig or fomites. 

Invaded hairs may show a sparse ectothrix or endothrix infection but do not fluoresce 

under Wood’s ultra-violet light. The geographical distribution is world-wide.

96 


Microsporum persicolor (Sabouraud) Guiart & Grigorakis 

Teleomorph: Arthroderma persicolor (Stockdale) Weitzman et al. 

Colonies (SDA) are generally flat, white to pinkish in colour, with a suede-like to granular 

texture and peripheral fringe. Reverse pigmentation is orange to red. Macroconidia 

are thin-walled, cigar-shaped, four- to seven-celled, 40-60 x 6-8 µm but are only rarely 

produced. Microconidia are abundant, spherical to pyriform. RG-2 organism. 

Key Features: microscopic morphology and culture characteristics. 

Culture and microconidia of Microsporum persicolor. 

20 µm 

Microsporum persicolor is a zoophilic fungus often occurring as a saprophyte on voles 

and bats. A rare cause of tinea corporis in humans. Not known to invade hair in 

vivo, but produces hair perforations in vitro. Distribution: Africa, Australia, Europe and 

North America.


Mortierella wolfii Mehrotra & Baijal 

The genus Mortierella has now been placed in a separate order, the Mortierellales 

(Cavalier-Smith 1998), and the genus contains about 90 recognised species, however 

Mortierella wolfii is probably the only pathogenic species being an important causal 

agent of bovine mycotic abortion, pneumonia and systemic mycosis in New Zealand, 

Australia, Europe and USA. 

Cultures are fast growing, white to greyish white, downy, often with a broadly zonate 

or lobed (rosette-like) surface appearance and no reverse pigment. Sporangiophores 

are typically erect, delicate, 80-250 µm in height, 6-20 µm wide at the base, arising 

from rhizoids or bulbous swellings on the substrate hyphae and terminating with 

a compact cluster of short acrotonous (terminal) branches. Sporangia are usually 

15-48 µm in diameter, with transparent walls and a conspicuous collarette is usually 

present following dehiscence of the sporangiospores. Columella are generally lacking 

and sporangiospores are single-celled, short-cylindrical, 6-10 x 3-5 µm, with a double 

membrane. Chlamydospores with or without blunt appendages (amoeba-like) may be 

present, zygospores have not been observed. Temperature: grows well at 40-42 O C; 

maximum 48 O C. RG-2 organism. 

Key Features: zygomycete, rapid growth at 40 O C (thermotolerant), and characteristic 

delicate acrotonous branching sporangia without columellae. 


(1980), McGinnis (1980), Rippon (1988), Smith (1989) and de Hoog et al. (2000). 

20 µm 

20 µm 

Culture of M. wolfii showing a broadly zonate or lobed rosette-like surface appearance 

and sporangium, showing a sporangiophore, wide at the base, arising from rhizoids, 

and acrotonous (terminal) branches, collarettes and sporangiospores.

98 


Mucor Micheli ex Staint-Amans 

The genus Mucor can be differentiated from Absidia, Rhizomucor and Rhizopus by the 

absence of stolons and rhizoids. Colonies are very fast growing, cottony to fluffy, white 

to yellow, becoming dark-grey, with the development of sporangia. Sporangiophores 

are erect, simple or branched, forming large (60-300 µm in diameter), terminal, globose 

to spherical, multispored sporangia, without apophyses and with well-developed 

subtending columellae. A conspicuous collarette (remnants of the sporangial wall) is 

usually visible at the base of the columella after sporangiospore dispersal. Sporangiospores 

are hyaline, grey or brownish, globose to ellipsoidal, and smooth-walled or 

finely ornamented. Chlamydospores and zygospores may also be present. 

Key Features: zygomycete, large, spherical, non-apophysate sporangia with pronounced 

columellae and conspicuous collarette at the base of the columella following 

sporangiospore dispersal. 

Antifungal 

20 µm 20 µm 

Sporangia, columella with a conspicuous collarette (arrow) 

and sporangiospores of Mucor. 

MIC µg/mL 

MIC µg/mL 

Range MIC 

Antifungal 







(2005), Sabatelli et al. (2006) and WCH in-house data.


Mucor Micheli ex Staint-Amans 

The genus Mucor contains about 50 recognised taxa, many of which have widespread 

occurrence and are of considerable economic importance (Zycha et al. 1969, Schipper 

1978, Domsch et al. 1980). However, only a few thermotolerant species are of 

medical importance and human infections are only rarely reported. Most infections 

reported list M. circinelloides and similar species such as M. indicus (M. rouxii), M. ramosissimus 

and M. amphibiorum as the causative agents. However, M. hiemalis and 

M. racemosus have also been reported as infectious agents, although their inability to 

grow at temperatures above 32 O C raises doubt as to their validity as human pathogens 

and their pathogenic role may be limited to cutaneous infections (Scholer et al. 1983, 

Goodman and Rinaldi 1991, Kwon-Chung and Bennett 1992, de Hoog et al. 2000). 

Maximum temperature for growth of the reported pathogenic species of Mucor. 

Species Max temp. ( O C) Pathogenicity 

M. amphibiorum 36 Animals, principally amphibians 

M. circinelloides 36-40 Animals, occasionally humans 

M. hiemalis 30 Questionable cutaneous infections only 

M. indicus 42 Humans and animals 

M. racemosus 32 Questionable 

M. ramosissimus 36 Humans and animals 

For descriptions of species, keys to taxa and additional information see Schipper 

(1978), Domsch et al. (1980), McGinnis (1980), Onions et al. (1981), Scholer et al. 

(1983), Rippon (1988), Goodman and Rinaldi (1991), Samson et al. (1995), de Hoog 

et al. (2000), Schipper and Staplers (2003) and Ellis (2005b). 

Mucor amphibiorum Schipper 

Colonies are greyish-brown, slightly aromatic and do not grow at 37 O C (maximum 

temperature for growth is 36 O C). Sporangiophores are hyaline, erect and mostly unbranched, 

rarely sympodially branched. Sporangia are dark-brown, up to 75 µm in 

diameter, and are slightly flattened with a diffluent membrane. Columellae are subglobose 

to ellipsoidal or pyriform, up to 60 x 50 µm, with small collarettes. Sporangiospores 

are smooth-walled, spherical, and 3.5-5.5 µm in diameter. Zygospores, 

when formed by compatible mating types, are spherical to slightly compressed, up to 

70 x 60 µm in diameter, with stellate projections. Mucor amphibiorum is distinguished 

by poor ramification of the sporangiophores and by globose sporangiospores. Ethanol 

and nitrate are not assimilated (Schipper 1978, Scholer et al. 1983, Hoog et al. 2000). 

RG-2 organism.

100 


Mucor circinelloides v. Tiegh 

M. circinelloides is a common and variable species that includes 4 forms: circinelloides, 

lusitanicus, griseo-cyanus and janssenii (Schipper 1978, Scholer et al. 1983). Colonies 

are floccose, pale greyish-brown and grow poorly at 37 O C (maximum growth temperature 

36-40 O C). Sporangiophores are hyaline and mostly sympodially branched with 

long branches erect and shorter branches becoming circinate (recurved). Sporangia 

are spherical, varying from 20-80 µm in diameter, with small sporangia often having a 

persistent sporangial wall. Columellae are spherical to ellipsoidal and are up to 50 µm 

in diameter. Sporangiospores are hyaline, smooth-walled, ellipsoidal, and 4.5-7 x 3.5- 

5 µm in size. Chlamydospores are generally absent. Zygospores are only produced 

in crosses of compatible mating types and are reddish-brown to dark-brown, spherical 

with stellate spines, up to 100 µm in diameter and have equal to slightly unequal suspensor 

cells. M. circinelloides differs from other species of Mucor in its formation of 

short circinated (coiled), branched sporangiophores bearing brown sporangia and its 

ability to assimilate ethanol and nitrate (Schipper 1976, Scholer et al. 1983, Samson et 

al. 1995, de Hoog et al. 2000, Schipper and Staplers 2003). RG-1 organism. 

Mucor indicus Lendner 

Colonies are characteristically deep-yellow, aromatic and have a maximum growth 

temperature of 42 O C. Sporangiophores are hyaline to yellowish, erect or rarely circinate 

and repeatedly sympodially branched, with long branches. Sporangia are yellow 

to brown, up to 75 µm in diameter, with diffluent membranes. Columellae are subglobose 

to pyriform, often with truncate bases, up to 40 µm high. Sporangiospores are 

smooth-walled, subglobose to ellipsoidal, and 4-5 µm in diameter. Chlamydospores 

are produced in abundance, especially in the light. Zygospores when formed by crosses 

of compatible mating strains are black, spherical up to 100 µm in diameter, with 

stellate spines and unequal suspensor cells. Mucor indicus differs from other species 

of Mucor by its characteristic deep-yellow colony colour, growth at over 40 O C, assimilating 

ethanol, but not nitrate, and being thiamine dependent (Schipper 1978, de Hoog 

et al. 2000, Schipper and Staplers 2003). RG-1 organism. 

Mucor ramosissimus Samutsevich 

Colonial growth is restricted, greyish and does not grow at 37 O C (maximum temperature 

for growth is 36 O C). Sporangiophores are hyaline, slightly roughened, tapering 

towards the apex and are erect with repeated sympodial branching. Sporangia are 

grey to black, globose or somewhat flattened, up to 80 µm in diameter and have very 

persistent sporangial walls. Columellae are applanate (flattened), up to 40-50 µm in 

size and are often absent in smaller sporangia. Sporangiospores are faintly brown, 

smooth-walled, subglobose to broadly ellipsoidal, 5-8 x 4.5-6 µm in size. Oidia may be 

present in the substrate hyphae, chlamydospores and zygospores are absent. Assimilation 

of ethanol is negative and that of nitrate is positive. Mucor ramosissimus differs 

from other species of Mucor by its low, restricted growth on any medium, extremely persistent 

sporangial walls, columellae that are applanate or absent in smaller sporangia 

(often resembling Mortierella species), short sporangiophores that repeatedly branch 

sympodially as many as 12 times, and the occurrence of racket-shaped enlargements 

in the sporangiophores (Hesseltine and Ellis 1964, Schipper 1976, Scholer et al. 1983, 

de Hoog et al. 2000, Schipper and Staplers 2003). RG-1 organism.


Nattrassia mangiferae (H. Syd. & Syd) Sutton & Dyko 

Pycnidial synanamorph: Hendersonula toruloidea Nattras 

Arthroconidial synanamorph: Scytalidium dimidiatum (Penzig) Sutton & Dyko 

Scytalidium hyalinum Campbell & Mulder 

This coelomycete is characterised by the presence of black, ostiolate pycnidia containing 

numerous hyaline, flask-shaped phialides. Phialoconidia are at first one-celled and 

hyaline, later becoming three-celled and brown, with the centre cell darker than the end 

cells. Conidia are ovoid to ellipsoidal in shape. Pycnidia are only occasionally formed 

in older cultures. In culture the Scytalidium dimidiatum anamorph is typically present 

showing chains of one- to two-celled, darkly pigmented arthroconidia, 3.5-5 x 6.5-12 

µm, produced by the holothallic fragmentation of undifferentiated hyphae. Cultures are 

effuse, hairy, dark grey to blackish-brown, or white to greyish, with a cream-coloured to 

deep ochraceous-yellow colony reverse. Colourless (hyaline) mutants (=Scytalidium 

hyalinum) often occur. 

Nattrassia mangiferae (arthric synanamorph = Scytalidium dimidiatum) is a recognised 

agent of onychomycosis and superficial skin infections, especially in tropical regions. 

However, isolates are sensitive to cycloheximide (actidione) which is commonly added 

to primary isolation media used for culturing skin scrapings. RG-2 organism. 


(1980), Moore (1986), Rippon (1988), Frankel and Rippon (1989), Sutton and Dyko 


20 µm 

Microscopic morphology of the Scytalidium dimidiatum synanamorph of Nattrassia 

mangiferae showing chains of one- to two-celled, darkly pigmented 

arthroconidia. 

Antifungal 




Amphotericin B 0.125-2 Itraconazole 0.03-32 Voriconazole 0.03-0.5 


Espinel-Ingroff et al. (2001) and WCH in-house data.

102 


Ochroconis gallopava (W.B. Cooke) de Hoog 

Colonies are smooth to suede-like, dry, flat, tobacco-brown to brownish-black with a 

dark brown diffusible pigment. Hyphae are brown with relatively thick walls. Conidiophores 

are mostly cylindrical to acicular, sometimes poorly differentiated, bearing a few 

conidia at the tip. Conidia are two-celled, subhyaline to pale brown, smooth-walled to 

verrucose, cylindrical to clavate, constricted at the septum, 11-18 x 2.5-4.5 µm in size, 

with the apical cell wider than the basal cell. A remnant of a denticle may also be seen 

at the conidial base. Optimum growth at 35 O C, tolerant to 40 O C. RG-2 organism. 

Ochroconis gallopava is a well recognised species and has been reported as an 

avian pathogen, especially in poultry. Occasional human infections have also been 

reported. 


1980, McGinnis 1980 and de Hoog et al. 2000. 

Antifungal 

10 µm 

10 µm 

Hyphae, conidiophores and conidia of Ochroconis gallopava. 




Amphotericin B 0.03-2 Itraconazole 0.03-0.5 Voriconazole 0.03-1 


McGinnis and Pasarell (1998), Espinel-Ingroff (2001) and WCH in-house data.


Onychocola canadensis Sigler 

Teleomorph: Arachnomyces nodososetosus Sigler & Abbott 

Colonies grow slowly and are velvety to lanose, white to yellowish, with a brownish 

reverse. Arthroconidia are cylindrical to broadly ellipsoidal, one- or two-celled, hyaline 

to subhyaline, 4-16 x 2-5 µm in size, forming long chains. Older cultures may show 

broad, brown, rough-walled hyphae. RG-2 organism. 

Onychocola canadensis is an uncommon cause of distal and lateral subungual or white 

superficial onychomycosis. However, it may sometimes be present in an abnormalappearing 

nail as an insignificant finding, not acting as a pathogen. 

Key Features: slow growing, white, arthroconidial mould isolated from nails. 

For descriptions of species, keys to taxa and additional information see Sigler and 

Congly (1990), Sigler et al. (1994), Gupta et al. (1998) and de Hoog et al. (2000). 

Culture and arthroconidia of Onychocola canadensis. 

10 µm

104 


Paecilomyces Bain 

Colonies are fast growing, powdery or suede-like, gold, green-gold, yellow-brown, lilac 

or tan, but never green or blue-green as in Penicillium. Phialides are swollen at their 

bases, gradually tapering into a rather long and slender neck, and occur solitarily, in 

pairs, as verticils, and in penicillate heads. Long, dry chains of single-celled, hyaline 

to dark, smooth or rough, ovoid to fusoid conidia are produced in basipetal succession 

from the phialides. 

The genus Paecilomyces may be distinguished from the closely related genus Penicillium 

by having long slender divergent phialides and colonies that are never typically 

green. Paecilomyces species are common environmental moulds and are seldom 

associated with human infection. However, some species, P. variotii, P. marquandii 

and P. lilacinus are emerging as causative agents of mycotic keratitis and of hyalohyphomycosis 

in the immunocompromised patient. 

Key Features: long slender divergent phialides and culture pigmentation. 

For descriptions of species, keys to taxa and additional information see Samson (1974), 

Domsch et al. (1980), McGinnis (1980), Onions et al. (1981), Rippon (1988) and de 

Hoog et al. (2000). 

a b 

Cultures of P. variotii (a) and P. lilacinus (b) showing colony pigmentation.


Antifungal 

Paecilomyces lilacinus (Thom) Samson 

Colonies are fast growing, suede-like to floccose, vinaceous to violet-coloured. Conidiophores 

are erect 400-600 µm in length, bearing branches with densely clustered 

phialides. Conidiophore stipes are 3-4 µm wide, yellow to purple and rough-walled. 

Phialides are swollen at their bases, gradually tapering into a slender neck. Conidia 

are ellipsoidal to fusiform, smooth-walled to slightly roughened, hyaline to purple in 

mass, 2.5-3.0 x 2-2.2 µm, and are produced in divergent chains. Chlamydospores are 

absent. Growth at 38 O C. RG-1 organism. 

Key Features: colony pigmentation, phialides with swollen bases, and pigmented and 

rough-walled conidiophore stipes, absence of chlamydospores. 

10 µm 10 µm 

Conidiophores, phialides and conidia of Paecilomyces lilacinus. Note: 

rough-walled conidiophore (arrow). 

MIC µg/mL 

MIC µg/mL 

Range MIC90 Antifungal Range MIC90 Amphotericin B 2-16 8 Posaconazole 0.06-2 0.5 (2) 

Itraconazole 0.5-16 16 Voriconazole 0.06-4 0.25 (2) 


Espinel-Ingroff et al. (2001), Espinel-Ingroff (2001, 2003), Gonzales et al. (2005), and 

WCH in-house data.

106 


Paecilomyces variotii Bain 

Colonies are fast growing, powdery to suede-like, funiculose or tufted, and yellowbrown 

or sand-coloured. Conidiophores bearing dense, verticillately arranged branches 

bearing phialides. Phialides are cylindrical or ellipsoidal, tapering abruptly into a 

rather long and cylindrical neck. Conidia are subspherical, ellipsoidal to fusiform, hyaline 

to yellow, smooth-walled, 3-5 x 2-4 µm and are produced in long divergent chains. 

Chlamydospores are usually present, singly or in short chains, brown, subspherical to 

pyriform, 4-8 µm in diameter, thick-walled to slightly verrucose. RG-2 organism. 

Key Features: yellow-brown colony pigmentation, cylindrical phialides, and presence 

of chlamydospores. 

10 µm 5 µm 10 µm 

Conidiophores, phialides, conidia and terminal chlamydospores of P. variotii. 

Antifungal 

MIC µg/mL 

MIC µg/mL 

Range MIC90 Antifungal Range MIC90 Amphotericin B 0.06-1 1 Posaconazole 0.03-0.5 0.5 

Itraconazole 0.03-8 0.5 Voriconazole 0.03-2 0.5 


Espinel-Ingroff et al. (2001), Espinel-Ingroff (2003) and WCH in-house data.


Paracoccidioides brasiliensis (Splendore) Almeida 

Colonies (SDA) at 25 O C are slow growing and variable in morphology. Colonies may 

be flat, wrinkled and folded, glabrous, suede-like or downy in texture, white to brownish 

with a tan or brown reverse. Microscopically, a variety of conidia may be seen, including 

pyriform microconidia, chlamydospores and arthroconidia. However, none of these 

are characteristic of the species, and most strains may grow for long periods of time 

without the production of conidia. 

On BHI blood agar at 37 O C, the mycelium converts to the yeast phase and colonies 

are white to tan, moist and glabrous and become wrinkled, folded and heaped. Microscopically, 

numerous large, 20-60 μm, round, narrow base budding yeast cells are 

present. Single and multiple budding occurs, the latter are thick-walled cells that form 

the classical “steering wheel” or “mickey mouse” structures that are diagnostic for this 

fungus, especially in methenamine silver stained tissue sections. 

WARNING: RG-3 Organism. Cultures of Paracoccidioides brasiliensis may represent 

a biohazard to laboratory personnel and should be handled with extreme caution in an 

appropriate pathogen handling cabinet. P. brasiliensis is geographically restricted to 

areas of South and Central America (Rippon, 1988). 

Key Features: clinical history, tissue pathology, culture identification with conversion 

to yeast phase at 37 O C. 



20 µm 

20 µm 

Multiple, narrow base budding yeast cells “steering wheels” of P. brasiliensis. 

Antifungal 

MIC µg/mL 


MIC µg/mL 

Range MIC90 Amphotericin B 0.03-4 0.25 Itraconazole 0.03-1 0.06 

Fluconazole 0.125-64 na Voriconazole 0.03-2 na 

Limited data available. Espinel-Ingroff et al. (2001), Espinel-Ingroff (2001) and 

Sabatelli et al. (2006) (na = not available).

108 


Penicillium Link: Fries 

Colonies are usually fast growing, in shades of green, sometimes white, mostly consisting 

of a dense felt of conidiophores. Microscopically, chains of single-celled conidia 

are produced in basipetal succession from a specialised conidiogenous cell called 

a phialide. The term basocatenate is often used to describe such chains of conidia 

where the youngest conidium is at the basal or proximal end of the chain. In Penicillium, 

phialides may be produced singly in groups or from branched metulae, giving a 

brush-like appearance (a penicillus). The penicillus may contain both branches and 

metulae (penultimate branches which bear a whorl of phialides). All cells between the 

metulae and the stipes of the conidiophores are referred to as branches. The branching 

pattern may be either simple (non-branched or monoverticillate), one-stage branched 

(biverticillate-symmetrical), two-stage branched (biverticillate-asymmetrical) or three- 

to more-staged branched. Conidiophores are hyaline, smooth or rough-walled. Phialides 

are usually flask-shaped, consisting of a cylindrical basal part and a distinct neck, 

or lanceolate (more or less with a narrow basal part tapering to a somewhat pointed 

apex). Conidia are in long dry chains, divergent or in columns, are globose, ellipsoidal, 

cylindrical or fusiform, hyaline or greenish, smooth or rough-walled. Sclerotia are 

produced by some species. RG-1 organisms, with the exception of P. marneffei which 

is an RG-3 organism. 

For identification, isolates are usually inoculated at three points on Czapek Dox agar 

and 2% Malt extract agar and incubated at 25 O C. Most species sporulate within 7 

days. Microscopic mounts are best made using a cellotape flag or a slide culture 

preparation mounted in lactophenol cotton blue. A drop of alcohol is usually needed to 

remove bubbles and excess conidia (see Samson et al., 1995). 

Key Features: hyphomycete, flask-shaped phialides arranged in groups from branched 

metulae forming a penicillus. 

metulae 

branches 

phialides 

a b c d 

Morphological structures and types of conidiophore branching in Penicillium. 

(a) simple; (b) one-stage branched; (c) two-stage branched; (d) three-stage 

branched (see Samson et al. 1995).


Penicillium Link:Fries 

Many species of Penicillium are common contaminants on various substrates and are 

known as potential mycotoxin producers. Correct identification is therefore important 

when studying possible Penicillium contamination of food. In some species odour and 

exudate production will help to recognise the taxa, but it should be pointed out that 

inhalation of conidia and volatiles may affect health. Human pathogenic species are 

rare, however opportunistic infections leading to mycotic keratitis, otomycosis and endocarditis 

(following insertion of valve prosthesis) have been reported (see Samson et 

al., 1995 and Rippon, 1988). 

For descriptions of species, keys to taxa and additional information see Raper and 

Thom (1949), Pitt (1979), Domsch et al. (1980), McGinnis (1980), Onions et al. (1981), 

Ramirez (1982), Samson et al. (1995) and de Hoog et al. (2000). 

20 µm 20 µm 

Conidiophores of P. verrucosum var. cyclopium showing two-stage branching. Simple 

conidiophore of P. cheresanum showing long chains of single-celled conidia. 

Antifungal 

MIC µg/mL 

MIC µg/mL 

Range MIC90 Antifungal Range MIC90 Amphotericin B 0.125-2 1-2 Posaconazole 0.03-2 0.25-1 

Itraconazole 0.03-2 0.5-2 Voriconazole 0.03->8 0.5-2 


Pfaller et al. (2002), Diekema et al. (2003), Espinel-Ingroff (2003) and WCH in-house 

data.

110 


Penicillium marneffei Segretain 

Colonies (SDA) at 25 O C are fast growing, suede-like to downy, white with yellowishgreen 

conidial heads. Colonies become greyish-pink to brown with age and produce 

a diffusible brownish-red to wine-red pigment. Conidiophores are hyaline, smoothwalled 

and bear terminal verticils of 3-5 metulae, each bearing 3-7 phialides. Conidia 

are globose to subglobose, 2 to 3 µm in diameter, smooth-walled and are produced in 

basipetal succession from the phialides. 

On brain heart infusion (BHI) blood agar incubated at 37 O C, colonies are rough, glabrous, 

tan-coloured and yeast-like. Microscopically, yeast cells are spherical to ellipsoidal, 

2-6 µm in diameter, and divide by fission rather than budding. Numerous short 

hyphal elements are also present. 

WARNING: RG-3 organism. Cultures of Penicillium marneffei may represent a biohazard 

to laboratory personnel and should be handled with caution in an appropriate 

pathogen handling cabinet. P. marneffei exhibits thermal dimorphism and is endemic 

in Southeast Asia and the southern region of China. 

Tissue sections show small, oval to elliptical yeast-like cells, 3 µm in diameter, either 

packed within histiocytes or scattered through the tissue. Occasional, large, elongated 

sausage-shaped cells, up to 8 µm long, with distinctive septa may be present. 

5 µm 

15 µm 5 µm 

Colony, a giemsa stained touch smear showing typical septate yeast-like cells 

(arrow), phialides and conidia of Penicillium marneffei.


Phaeoacremonium parasiticum (Ajello et al.) W. Gams et al. 

Synonym: Phialophora parasiticum Ajello, Gerog & Wang 

Cultures are usually slow growing, suede-like with radial furrows, initially whitish-grey 

becoming olivaceous-grey with age. Hyphae hyaline, later becoming brown and some 

becoming rough-walled. Phialides are brown, thick-walled, slender, acular to cylindrical 

slightly tapering towards the tip, 15-50 μm long, often proliferating, with small, funnel-shaped 

collarettes. Conidia, often in balls, are hyaline, thin-walled, cylindrical to 

sausage-shaped, 3-6 x 1-2 μm, later inflating (de Hoog et al. 2000). RG-2 organism. 

P. parasiticum is a plant pathogen but it has also been reported from cases of 

subcutaneous infection, arthritis, mycetoma, endocarditis and mycotic keratitis. 

For descriptions of species and additional information see de Hoog et al. (2000). 

Antifungal 

20 µm 

10 µm 

Colony, phialides and conidia of Phaeoacremonium parasiticum. 







112 


Phialophora Medlar 

Colonies are usually slow growing, grey to olivaceous-black, often becoming brown 

with age. Microscopically, members of the genus Phialophora produce clusters of 

single-celled conidia in basipetal succession from characteristic flask-shaped or cylindrical 

phialides which have distinctive collarettes. Conidia are hyaline to olivaceous 

brown, smooth-walled, ovoid to cylindrical or allantoid, and usually aggregate in slimy 

heads at the apices of the phialides, which may be solitary, or in a brush-like arrangement. 

The genus Phialophora contains more than 40 species, most are saprophytes commonly 

found in soil or on decaying wood. However, several species have been documented 

as causing either chromoblastomycosis (P. verrucosa) or phaeohyphomycosis 

(P. verrucosa and P. richardsiae). 

Phialophora richardsiae (Nannf.) Conant 

Colonies grow rapidly, and are powdery to woolly or tufted, greyish-brown with a greybrown 

to olivaceous-black reverse. Two conidial types are produced; (1) hyaline conidia 

which are allantoid or cylindrical, 3-6 x 1.5-2.5 μm in size, formed on inconspicuous, 

peg-like phialides on thin-walled hyphae; and (2) brown, thick-walled conidia which are 

spherical to sub-spherical, 2.5-3.5 x 2-3 μm, formed on dark brown, slender, tapering 

phialides with flaring collarettes. RG-2 organism. 

Phialophora richardsiae is a soft rot fungus of wood and is an uncommon cause of 

human infection. However, cases of subcutaneous phaeohyphomycosis have been 

reported. 

Antifungal 

10 µm 

10 µm 

Phialides of P. richardsiae producing 2 types of conidia. (1) hyaline conidia, formed on 

inconspicuous, peg-like phialides on thin-walled hyphae; and (2) brown, thick-walled 

conidia formed on dark brown, slender, tapering phialides with flaring collarettes. 








Phialophora verrucosa Medlar 

Colonies (SDA) are slow growing, initially dome-shaped, later becoming flat, suedelike 

and olivaceous to black in colour. Phialides are flask-shaped or elliptical with distinctive 

funnel-shaped, darkly pigmented collarettes. Conidia are ellipsoidal, smoothwalled, 

hyaline, mostly 3.0-5.0 x 1.5-3.0 μm, and aggregate in slimy heads at the 

apices of the phialide. RG-2 organism. 

Phialophora verrucosa is a well documented causative agent of chromoblastomycosis, 

and mycetoma. It produces characteristic flask-shaped phialides with distinctive funnel-shaped, 

darkly pigmented collarettes. 


McGinnis (1978), Domsch et al. (1980), McGinnis (1980) and de Hoog et al. (2000). 

Antifungal 

20 µm 

20 µm 

Phialides and conidia of Phialophora verrucosa. 







114 


Phoma Saccardo 

Colonies are spreading, greyish-brown, powdery or suede-like and produce large, 

globose, membranous to leathery, darkly pigmented, ostiolate pycnidia. Conidia are 

produced in abundance within the pycnidia on narrow thread-like phialides, which are 

hardly differentiated from the inner pycnidial wall cells. Conidia are globose to cylindrical, 

one-celled, hyaline, and are usually extruded in slimy masses from the apical 

ostiole. RG-1 organism. 

Members of the genus Phoma have a world-wide distribution and are ubiquitous in 

nature, with over 2000 species having been described from soil, as saprophytes on 

various plants, and as pathogens to plants and humans. 

Key Features: coelomycete, ostiolate pycnidia producing masses of slimy, hyaline, 

single-celled conidia. 

For descriptions of species, keys to taxa and additional information see Punithalingam 

(1979), McGinnis (1980), Sutton (1980), Rippon (1988), Montel et al. (1991), Samson 

et al. (1995) and de Hoog et al. (2000). 

20 µm 

Pycnidia of Phoma.


Pithomyces Berkeley and Broome 

Colonies are fast growing, dark grey to black, suede-like to downy and produce darkly 

pigmented, multicellular conidia formed on small peg-like branches of the vegetative 

hyphae. Conidia are broadly elliptical, pyriform, oblong, and are commonly echinulate 

or verrucose. 

The genus Pithomyces contains about 15 species commonly found from a very wide 

range of plant material, also from air, soil, hay, sawn timber and ceiling plaster. However, 

one species, Pithomyces chartarum is often involved with facial eczema of sheep. 

Pithomyces chartarum (Berk. & M.A. Curtis) M.B. Ellis 

Colonies are fast growing, suede-like to downy and black. Conidiophores are pale olive, 

smooth or verrucose, 2.5-10 x 2-3.5 µm. Conidiogenous cells integrated, intercalary 

or terminal, indeterminate, with 1-2 loci of similar width in the conidiogenous cells. 

Conidia muriform, medium to dark brown, echinulate to verrucose, 3 (-4)-euseptate, 

slightly constricted at the septa, with one or both median cells divided by longitudinal 

septa, thick-walled, broadly ellipsoidal, apex obtuse, base truncate and characteristically 

with part of the conidiogenous cell remaining attached as a small pedicel, 18-29 

x 10-17 µm. RG-1 organism. 

Key Features: dematiaceous hyphomycete with multi-celled conidia produced on 

small peg-like branches of the vegetative hyphae. 


1976), Domsch et al. (1980) and Rippon (1988). 

15 µm 

Conidiophores and conidia of Pithomyces chartarum.

116 


Prototheca Kruger 

Prototheca species are achlorophyllous algae with phylogenetic affinities to the genus 

Chlorella. Colonies are smooth, moist, white to cream and yeast-like. Cultures are 

sensitive to cycloheximide (actidione) and optimal growth occurs at 25 O C to 30 O C. 

Mycelium and conidia are absent. Vegetative cells are globose to ovoid, hyaline, varying 

in size from approximately 8-20 µm, and have a relatively thick and highly refractile 

wall. No budding cells are present; reproduction is by the development of large sporangia 

(theca) which contain from 2-20 or more small sporangiospores (endospores or 

autospores) which are asexually produced by nuclear division and cleavage of the cytoplasm 

(Kaplan, 1977, McGinnis, 1980, Rippon, 1988, Pore, 1985). RG-1 organism. 

Key Features: achlorophyllous algae reproducing by sporangia (theca) and sporangiospores 

(autospores). The genus Prototheca contains four species which can be 

differentiated by assimilation tests and morphological criteria as outlined below. The 

API 20C yeast identification strip may be used for species identification. So far only P. 

wickerhamii and P. zopfii have been involved in human or animal infections. 

P. wickerhamii P. zopfii P. stagnora P. moriformis 

Growth at 37 O C + + - - 

Glucose + + + + 

Trehalose + - - - 

L-propanol - + +/- + 

Acetate (pH5) - + +/- + 

Galactose + - + - 

Capsule - - + + 

Antifungal 

5 µm 

Thecae and autospores of Prototheca wickerhamii. 


Antifungal 

Antifungal 


Amphotericin B 0.25-0.5 Voriconazole 0.25 Posaconazole 0.25 



Ramichloridium Stahel ex de Hoog 

Colonies are rapid growing, smooth, farinose or velvety, brown or olivaceous-green, 

often with orange or yellow soluble pigments. Conidiophores are erect, brown, apically 

with small denticles on which conidia are produced in sympodial succession. Conidia 

are one-celled, hyaline to pale brown. 

Ramichloridium contains about 25 species that are usually associated with forest litter 

and rotting wood, however the genus contains two species of medical interest; R. 

mackenziei and R. schulzeri. 

Ramichloridium schulzeri (Sacc.) de Hoog 

Colonies growing moderately rapidly, consisting of a rather compact, flat, submerged 

mycelium, pale orange, locally with some powdery, brownish aerial mycelium; reverse 

pink to orange. Conidiophores are erect, straight, unbranched, thick-walled, reddishbrown, 

up to 250 µm high, gradually becoming paler towards the apex, of variable 

length, elongating sympodially during conidiogenesis, with scattered, pimple-shaped 

conidium bearing denticles which have unpigmented scars. Conidia are subhyaline, 

smooth-walled or slightly rough-walled, ellipsoidal, obovoidal or fusiform, 6.5-10.0 x 3- 

4 µm, usually with an acuminate base and unpigmented scars. RG-1 organism. 

For descriptions of species, keys to taxa and additional information see de Hoog (1977), 

Rippon et al. (1985) and de Hoog et al. (2000). 

10 µm 

20 µm 

Conidiophores showing sympodial development of conidia of R. schulzeri.

118 


Rhinocladiella Nannfeldt 

Colonies are restricted, velvety, lanose or nearly smooth, grey to olivaceous-brown. 

Hyphae pale olivaceous. Conidiophores are slightly differentiated, sub-erect, usually 

branched, pale to dark brown. Conidiogenous cells are intercalary or free, cylindrical, 

in the apical part with conidium bearing denticles with unpigmented scars. Conidia are 

hyaline to subhyaline, one-celled and smooth-walled. Budding cells and an accompanying 

Exophiala state may be present. 

Rhinocladiella contains 6-8 species, with two species of medical interest; R. atrovirens 

and R. aquaspersa. 

Rhinocladiella atrovirens Nannfeldt 

Colonies are restricted, velvety or lanose, olivaceous, often slightly mucoid at the centre; 

reverse dark olivaceous green to blackish. Conidiophores are short, brown, thickwalled. 

Conidiogenous cells are cylindrical, intercalary or free, 9-19 x 1.6-2.2 µm; 

denticulate rachis up to 15 µm long, with crowded, flat or butt-shaped, unpigmented 

conidial denticles. Conidia are hyaline, thin- and smooth-walled, short-cylindrical, 

with truncate basal scars, 3.7-5.5 x 1.2-1.8 µm. Budding cells, if present, are hyaline, 

thin-walled, broadly ellipsoidal, 3.0-4.3 x 1.7-2.5 µm. Germinating cells are inflated, 

spherical to subspherical, 4.5-6.0 µm. An annellidic Exophiala synanamorph may be 

present. RG-1 organism. 

For descriptions of species, keys to taxa and additional information see de Hoog (1977, 

1983), Schell et al. (1983) and de Hoog et al. (2000). 

Antifungal 

10 µm 10 µm 10 µm 

Culture, conidiophores showing a terminal denticulate rachis, conidia and budding 

yeast cells of Rhinocladiella atrovirens. 




Amphotericin B 0.03-0.25 Itraconazole 0.03-0.06 Voriconazole 0.03-0.5 




The genus Rhizomucor is distinguished from Mucor by the presence of stolons and 

poorly developed rhizoids at the base of the sporangiophores and by the thermophilic 

nature of its 3 species: R. miehei, R. pusillus and R. tauricus. All 3 of these species 

are potential human and animal pathogens and were originally classified in the genus 

Mucor. Rhizomucor variabilis as described by de Hoog et al. (2000) is not thermophilic 

and is probably a degenerate culture of Mucor hiemalis (Voigt et al., 1999). Rhizomucor 

pusillus is cosmopolitan and both R. miehei and R. pusillus have been reported as 

pathogens to humans and animals, the latter to a greater extent. 

For descriptions of species, keys to taxa and additional information see Cooney and 

Emerson (1964), Schipper (1978), Domsch et al. (1980), McGinnis (1980), Ellis and 

Keane (1981), Scholer et al. (1983), Hoog et al. (2000), Schipper and Staplers (2003) 

and Ellis (2005b). 

Remember, identification of most zygomycetes is based primarily on the morphology 

of the sporangia; i.e. arrangement and number of sporangiospores, shape, colour, 

presence or absence of columellae and apophyses, as well as the arrangement of the 

sporangiophores and the presence or absence of rhizoids. Growth temperature tests 

can also be especially helpful in identifying and differentiating members of the genera 

Rhizomucor, Rhizopus and Absidia. 

Rhizomucor miehei (Cooney and Emerson) Schipper 

Synonym: Mucor miehei Lindt 

This species has been reported as a rare cause of bovine mastitis (Scholer et al. 1983) 

and is similar in most respects to R. pusillus. However, all strains are homothallic 

forming numerous zygospores, which are reddish-brown to blackish-brown, globose to 

slightly compressed, up to 50 µm in diameter, with stellate warts and equal suspensor 

cells. Colony colour is a dirty grey rather than brown, and sporangia have spiny walls, 

are up to 50-60 µm in diameter, with columellae rarely larger than 30 µm in diameter. 

Growth is stimulated by thiamine, with no assimilation of sucrose and maximum temperature 

of growth is 54–58 O C. RG-1 organism. 

Key Features: growth at 45 O C, the formation of numerous zygospores, a dirty grey 

culture colour and a partial growth requirement for thiamine. 

Synonym: Mucor pusillus Lindt 

Rhizomucor Lucet & Costantin 

Rhizomucor pusillus (Lindt) Schipper 

This species is a rare human pathogen. It has been reported from cases of pulmonary, 

disseminated and cutaneous types of infection. It is more often associated with animal 

disease, especially bovine abortion. Rhizomucor pusillus has a world-wide distribution 

and is commonly associated with compost heaps.

120 


Rhizomucor pusillus (Lindt) Schipper 

This thermophilic zygomycete is readily recognizable by its characteristic compact, low 

growing (2-3 mm high), grey to greyish brown-coloured mycelium and by the development 

of typical sympodially branched, hyaline to yellow-brown sporangiophores (8-15 

μm in diameter), always with a septum below the sporangium. Sporangia are globose 

(40-60 µm in diameter), each possessing an oval or pear-shaped columella (20-30 

µm), often with a collarette. Sporangiospores are hyaline, smooth-walled, globose to 

subglobose, occasionally oval (3-5 µm), and are often mixed with crystalline remnants 

of the sporangial wall. Chlamydospores are absent. Zygospores are rough-walled, 

reddish brown to black, 45-65 mm in diameter and may be produced throughout the 

aerial hyphae in matings between compatible isolates. Temperature growth range: 

minimum 20-27 O C; optimum 35-55 O C; maximum 55-60 O C. There is positive assimilation 

of sucrose and no thiamine dependence. RG-2 organism. 

Key Features: zygomycete, growth at 45 O C (thermophilic), poorly developed stolons 

and rhizoids, branching sporangiophores with a septum below the sporangium, darkcoloured 

sporangia without apophyses and smooth-walled globose to subglobose 

sporangiospores. 

15 µm 20 µm 

Sporangiophores, collumellae and primitive rhizoids of R. pusillus. 

Antifungal 

MIC µg/mL 

MIC µg/mL 

Antifungal 

Range Range 

Fluconazole >64 Amphotericin B 0.06-0.25 


Posaconazole 0.06-0.25 Voriconazole 2-16 

Very limited data, antifungal susceptibility testing of individual strains is 

recommended. Dannaoui et al. (2003), Singh et al. (2005) and WCH inhouse 

data.


The genus Rhizopus is characterised by the presence of stolons and pigmented rhizoids, 

the formation of sporangiophores, singly or in groups from nodes directly above 

the rhizoids, and apophysate, columellate, multispored, generally globose sporangia. 

After spore release the apophyses and columella often collapse to form an umbrellalike 

structure. Sporangiospores are globose to ovoid, one-celled, hyaline to brown 

and striate in many species. Colonies are fast growing and cover an agar surface with 

a dense cottony growth that is at first white becoming grey or yellowish brown with 

sporulation. 

Antifungal 

Rhizopus Ehrenberg ex Corda. 

MIC µg/mL 

MIC µg/mL 

Range MIC 

Antifungal 







(2005), Sabatelli et al. (2006) and WCH in-house data. 

In the past, numerous attempts have been made to clarify the species concepts of 

the genus Rhizopus. Recently, 3 excellent revisions with easy to use keys have been 

produced by Schipper (1984), Ellis (1985, 1986) and Schipper and Stalpers (2003). 

Basically, 3 groups have been recognised: the ‘stolonifer’ group, the ‘oryzae’ group and 

the ‘microsporus’ group. The G-C values of the 3 groups have been defined by Frye 

and Reinhardt (1993), and temperature growth studies at 30 O C, 36 O C and 45 O C are 

characteristic for each of the groups. 

The ‘stolonifer’ group has sporangia up to 275 µm in diameter and grows at 30 O C, but 

has a maximum growth temperature of 36 O C. Species in this group include R. sexualis 

and R. stolonifer. The latter has been unconvincingly implicated in human infection 

(Ferry and Abedi 1983), although with a maximum growth temperature of only 32 O C its 

pathogenicity is thus questionable. 

The ‘oryzae’ group has been reduced to a single species that is able to grow at 40 O C 

but not at 45 O C, and has sporangia not exceeding 240 µm in diameter. There is no 

doubt that R. oryzae and R. arrhizus are synonymous, the contentious issue being 

which species name to use. The taxonomic treatment of Schipper and Stalpers (2003) 

will be used in this book; however, the synonym R. arrhizus is commonly used in the 

medical literature. Rhizopus oryzae is an important human pathogen. 

The ‘microsporus’ group has simple rhizoids, and smaller sporangia up to 100 µm in 

diameter and grows at both 40 and 45 O C. This group contains 4 species: R. homothallicus, 

R. azygosporus, R. schipperae and R. microsporus with the later subdivided into 

3 varieties, namely R. microsporus var. microsporus, R. microsporus var. oligosporus 

and R. microsporus var. rhizopodiformis. All are thermophilic and R. microsporus is a 

well-recognised pathogen of humans and animals.

122 


Rhizopus Ehrenberg ex Corda. 

Differentiation of pathogenic Rhizopus microsporus group isolates. 

Species Growth 

at 45O Growth 

C at 50O Main species characteristics 

C 

R. azygosporus Good No Abundant azygospores 

R. microsporus var. Good No Sporangiospores angular to ellipsoidal 

microsporus 

and distinctly striate, up to 5–6 µm diam 

R. microsporus var. Restricted No Sporangiospores globose, up to 

oligosporus 

9 µm diameter or more, heterogeneous 

R. microsporus var. Good Good Sporangiospores globose rarely over 5 

rhizopodiformis 

µm in diameter minutely spinulose 

R. schipperae Good No Abundant chlamydospores and 

restricted sporulation 

For descriptions of species, keys to taxa and additional information, see Domsch et al. 

(1980), McGinnis (1980), Onions et al. (1981), Scholer et al. (1983), Schipper (1984), 

Schipper and Stalpers (1984, 2003), Ellis (1985, 1986), Rippon (1988), Kwon-Chung 

and Bennett (1992), Samson et al. (1995); Hoog et al. (2000) and Ellis (2005b). 

Rhizopus azygosporus Yuan & Jong 

Rhizopus azygosporus is closely related to R. microsporus (Yuan and Jong 1984) and 

has been reported as the causative agent of 3 fatal cases of gastrointestinal infection 

in premature babies (Woodward et al. 1992, Schipper et al. 1996). Previously, this fungus 

was only known from its type culture, which has been isolated from tempeh, a solid 

fermented soybean food from Indonesia (Yuan and Jong 1984). Colonies are whitish 

to grey-black, producing pale-brown simple rhizoids. Sporangiophores are brownish, 

up to 350 µm high and 6-14 µm wide. Sporangia are greyish-black, spherical and 

50-100 µm in diameter. Columellae are subglobose to globose. Sporangiospores 

are ovoid to ellipsoidal 4-5 to 6-7 µm in diameter with faint striations. Azygospores 

are pale to dark-brown, spherical to subglobose, 30–70 µm in diameter, with coarse 

conical projections. All strains produce abundant azygospores in unmated isolates as 

a species characteristic. There is good growth at 45 O C with a maximum of 46-48 O C. 


Rhizopus microsporus var. microsporus 

Rhizopus microsporus var. microsporus is a rare cause of human infection (Kerr et al. 

1988, Kwon-Chung and Bennett, 1992). Colonies are pale brownish-grey producing 

simple rhizoids. Sporangiophores are brownish, up to 400 µm high and 10 µm wide, 

but most are smaller and are produced in pairs. Sporangia are greyish-black, spherical, 

up to 80 µm in diameter. Columellae are subglobose to globose to conical. Sporangiospores 

are angular to broadly ellipsoidal to lemon-shaped, quite equal in size, 

up to 5-6 µm in diameter and are distinctly striate. Zygospores, formed by crosses of 

compatible mating strains, are dark red–brown, spherical, up to 100 µm in diameter, 

with stellate projections and unequal suspensor cells. There is good growth at 45 O C, 

with a maximum of 46–48 O C. RG-2 organism.


Rhizopus microsporus var. oligosporus (Saito) Schipper & Stalpers 

Rhizopus microsporus var. oligosporus is a rare cause of human zygomycosis (Tintelnot 

and Nitsche 1989). Colonies are pale yellowish-brown to grey and sporulation is 

often poor. Rhizoids are subhyaline and simple. Sporangiophores are brownish, up to 

300 µm high and 15 µm wide, with 1-3 produced together. Sporangia are black, spherical, 

up to 100 µm in diameter. Columellae are subglobose to somewhat conical. Sporangiospores 

are subglobose to globose, up to 9 µm in diameter, almost smooth, with 

larger spores often irregular in shape. Chlamydospores are abundant, hyaline, single 

or in chains, spherical, ellipsoidal or cylindrical, 7-35 µm in diameter. Zygospores are 

not known. There is growth at 45 O C with a maximum of 46–48 O C. RG-2 organism. 

Rhizopus microsporus var. rhizopodiformis (Cohn) Schipper & Stalpers 

Rhizopus microsporus var. rhizopodiformis is the second most frequently isolated zygomycete, 

accounting for between 10% and 15% of reported human cases (Scholer 

et al. 1983; Kwon-Chung and Bennett 1992). Colonies are dark greyish-brown, up to 

10 mm high with simple rhizoids. Sporangiophores are brownish, up to 500 µm high 

and 8 µm wide, with 1-4 produced together. Sporangia are bluish to greyish-black, 

spherical and up to 100 µm in diameter. Columellae are pyriform comprising 80% of 

the sporangium. Sporangiospores are subglobose to globose, quite equal in size, up 

to 6 µm in diameter and minutely spinulose. Zygospores, when formed by crosses of 

compatible mating strains, are reddish-brown, spherical, up to 100 µm in diameter, with 

stellate projections and unequal suspensor cells. There is good growth at 45 O C with a 

maximum of 50–52 O C. RG-2 organism. 

30 µm 

Sporangia showing sporangiophores, columellae, sporangiospores 

and rhizoids of R. microsporus var. oligosporus.

124 


Rhizopus oryzae Went & Prinsen Geerligs 

Synonym: Rhizopus arrhizus Fischer 

Rhizopus oryzae is the most common causative agent of zygomycosis, accounting for 

some 60% of the reported culture positive cases, and nearly 90% of the rhinocerebral 

form of infection. Colonies are very fast growing, about 5-8 mm high, with some tendency 

to collapse, white cottony at first becoming brownish grey to blackish-grey depending 

on the amount of sporulation. Sporangiophores up to 1500 µm in length and 

18 µm in width, smooth-walled, non-septate, simple or branched, arising from stolons 

opposite rhizoids usually in groups of 3 or more. Sporangia are globose, often with a 

flattened base, greyish black, powdery in appearance, up to 175 µm in diameter and 

many spored. Columellae and apophysis together are globose, subglobose or oval, up 

to 130 µm in height and soon collapsing to an umbrella-like form after spore release. 

Sporangiospores are angular, subglobose to ellipsoidal, with ridges on the surface, 

and up to 8 µm in length. No growth at 45 O C; good growth at 40 O C. RG-2 organism 

20 µm 

100 µm 

Culture, sporangia, sporangiophores and rhizoids of R.oryzae.


Rhodotorula Harrison 

The genus Rhodotorula is characterised by the combination of red or yellow cultures 

due to the presence of carotenoid pigments, the inability to assimilate inositol and the 

absence of fermentation. The basidiomycetous nature of yeasts is usually indicated 

by a positive urease test. The genus Cryptococcus is similar to Rhodotorula both in 

production of carotenoid pigments and the presence of capsulated blastoconidia. The 

distinctive difference between the two is the assimilation of inositol, which is positive in 

Cryptococcus. 

Rhodotorula mucilaginosa is a common airborne contaminant of skin, lungs, urine and 

faeces. R. mucilaginosa is a known cause of fungal peritonitis in patients on continuous 

ambulatory peritoneal dialysis (CAPD). This is usually due to saprophytic colonisation 

of catheters or dialysis machinery and removal of the source of contamination 

usually leads to clearing of the symptoms. 


(1980), Barnett et al. (1983), Kreger-Van Rij (1984), Rippon (1988), Kurtzman and 

Fell (1988) and de Hoog et al. (2000). 

Culture of Rhodotorula mucilaginosa.

126 


Rhodotorula glutinis (Fresenius) Harrison 

Culture: Colonies (SDA) are coral red to salmon-coloured or slightly orange, smooth 

to wrinkled, highly glossy to semi-glossy. Mucoid to pasty to slightly tough, yeast-like 

colonies. 

Microscopy: Ovoidal to globose or more elongate budding yeast-like cells or blastoconidia, 

2.3-5.0 x 4.0-10.0 µm. 

India Ink Preparation: Small capsules present. 

Dalmau Plate Culture on Cornmeal and Tween 80 Agar: Budding blastoconidia 

only. No pseudohyphae are formed. 





Galactose - Cellobiose v L-Rhamnose v DL-Lactate v 

Sucrose - Trehalose + D-Glucosamine - myo-Inositol - 


Lactose - Melibiose - Glycerol v D-Glucuronate v 

Trehalose - Raffinose v Erythritol - Nitrate - 


Glucose + Soluble Starch - Galactitol v 0.1% Cycloheximide v 

Galactose v D-Xylose v D-Mannitol v Growth at 37 O C v 

Key Features: germ tube negative yeast and sugar assimilation pattern. Common 

saprophyte however cases of fungemia have been reported. RG-1 organism. 

Antifungal 

MIC µg/mL 

MIC µg/mL 


Range 

Fluconazole >64 Amphotericin B 0.06-0.25 

Itraconazole 0.5-1 Flucytosine 0.03-0.06 

Voriconazole 1-4 Caspofungin 0.25 




Synonym: Rhodotorula rubra (Demme) Lodder. 

Culture: Colonies (SDA) are coral pink, usually smooth, sometimes reticulate, rugose 

or corrugated, moist to mucoid, yeast-like colonies. 

Microscopy: Spherical to elongate budding yeast-like cells or blastoconidia, 2.5-6.5 x 

6.5-14.0 µm. 


Dalmau Plate Culture on Cornmeal and Tween 80 Agar: Budding blastoconidia 

only. No pseudohyphae are formed. 


saprophyte however cases of peritonitis and fungemia have been reported. RG-1 

organism. 

Antifungal 

Rhodotorula mucilaginosa (Jorgensen) Harrison 




Glucose - Maltose v D-Ribose v D-Gluconate + 

Galactose - Cellobiose v L-Rhamnose v DL-Lactate v 

Sucrose - Trehalose + D-Glucosamine v myo-Inositol - 



Trehalose - Raffinose + Erythritol v Nitrate - 

Assimilation Melezitose v Ribitol v Urease + 

Glucose + Soluble Starch - Galactitol v 0.1% Cycloheximide - 

Galactose v D-Xylose + D-Mannitol v Growth at 40 O C + 

MIC µg/mL 

MIC µg/mL 

Range MIC90 Antifungal Range MIC90 Fluconazole 0.5->64 >64 Amphotericin B 0.03-1 0.5 

Itraconazole 0.25-4 2 Flucytosine 0.03-0.25 0.25 

Voriconazole 0.25-4 2 Caspofungin 16 >16 


Espinel-Ingroff et al. (2001), Espinel-Ingroff (2003), Cuenca-Estrella et al. (2006) and 

WCH in-house data.

128 


Saccharomyces cerevisiae Meyen ex Hansen 

The genus Saccharomyces is characterised by strong fermentation of at least glucose 

and the presence of rather large globose to ellipsoidal cells with multilateral budding. 

Pseudohyphae can be formed, but never true hyphae. Ascospores are globose to ellipsoidal, 

with a smooth wall, usually one to four per ascus. Lactose and nitrate are 

not utilised. The species S. cerevisiae, commonly known as Baker’s yeast, is the most 

important representative. 

Culture: Colonies (SDA) are white to cream, smooth, glabrous yeast-like colonies. 

Microscopy: Large globose to ellipsoidal budding yeast-like cells or blastoconidia, 

3.0-10.0 x 4.5-21.0 µm. 


Dalmau Plate Culture on Cornmeal and Tween 80 Agar: Usually budding blastoconidia 

only, however pseudohyphae may be formed rarely. 



Fermentation Sucrose + D-Arabinose - α-M-D-glucoside v 

Glucose + Maltose + D-Ribose - D-Gluconate - 



Maltose v Lactose - N-A-D-glucosamine - 2-K-D-gluconate - 

Lactose - Melibiose v Glycerol - D-Glucuronate - 

Trehalose - Raffinose + Erythritol - Nitrate - 

Assimilation Melezitose v Ribitol - Urease - 


Galactose v D-Xylose - D-Mannitol - Growth at 37 O C v 


food and environmental saprophyte. RG-1 organism. 


(1980), Barnett et al. (1983), Kreger-Van Rij (1984), Rippon (1988), Kurtzman and 

Fell (1988) and de Hoog et al. (2000). 

Antifungal 

MIC µg/mL 

MIC µg/mL 

Range MIC90 Antifungal Range MIC90 Fluconazole 0.125-16 8 Amphotericin B 0.06-2 1 

Itraconazole 0.03-4 0.5 Flucytosine 0.03-0.5 0.25 

Voriconazole 0.06-0.25 0.125 Caspofungin 1 1 


Espinel-Ingroff et al. (2001), Espinel-Ingroff (2003), Cuenca-Estrella et al. (2006) 



Saksenaea vasiformis Saksena 

The genus Saksenaea is characterised by the formation of flask-shaped sporangia 

with columellae and simple, darkly pigmented rhizoids. Saksenaea vasiformis is the 

only known species and appears to have a world-wide distribution in association with 

soil. It is an emerging human pathogen (Holland, 1997) that is most often associated 

with cutaneous or subcutaneous lesions after trauma. Colonies are fast growing, 

downy, white with no reverse pigment made up of broad, non-septate hyphae typical of 

a zygomycetous fungus. Sporangia are typically flask-shaped with a distinct spherical 

venter and long-neck, arising singly or in pairs from dichotomously branched, darkly 

pigmented rhizoids. Collumellae are prominent and dome-shaped. Sporangiospores 

are small, oblong, 1-2 x 3-4 µm, and are discharged through the neck following the dissolution 

of an apical mucilaginous plug. RG-2 organism. 

Key Features: zygomycete, unique flask-shaped sporangia, failure to sporulate on 

primary isolation media. 

Laboratory identification of this fungus may be difficult or delayed because of the 

mould’s failure to sporulate on the primary isolation media or on subsequent subculture 

onto potato dextrose agar. Sporulation may be stimulated by the use of nutrient deficient 

media, like cornmeal-glucose-sucrose-yeast extract agar, Czapek Dox agar, or 

by using the agar block method described by Ellis and Ajello (1982), Ellis and Kaminski 

(1985) and Padhye and Ajello (1988). 

For descriptions of species, keys to taxa and additional information see Saksena 

(1953), Ellis and Hesseltine (1966), Ajello et al. (1976), Ellis and Ajello (1982), Ellis 

and Kaminski (1985), Pritchard et al. (1986), Padhye et al. (1988), Padhye and Ajello 

(1988), Goldschmied-Reouven et al. (1989), de Hoog et al. (2000) and Ellis (2005b). 

Antifungal 

The agar block method to induce 

sporulation of Saksenaea vasiformis 

and Apophysomyces elegans. 

A small block of agar is cut from a well 

established culture grown on PDA and 

is placed in the centre of petri dish 

containing 1% agar in distilled water. 

After 21 days at 26 O C look for sporangium 

formation at the periphery of the 

petri dish. 

MIC µg/mL 

MIC µg/mL 

Antifungal 

Range Range 

Fluconazole >64 Amphotericin B 0.125-2 


Posaconazole 0.015-0.25 Voriconazole 0.5-4 

Very limited data, poor sporulation, antifungal susceptibility testing of individual strains 

is recommended. Sun et al. (2002) and WCH in-house data.

130 


Saksenaea vasiformis Saksena 

15 µm 

Sporangium of Saksenaea vasiformis.


Scedosporium apiospermum/aurantiacum Complex 

Recent genetic studies have shown Scedosporium apiospermum to be a species complex 

and two species of medical importance have now been recognised: S. apiospermum 

and S. aurantiacum (Gilgado et al. 2005). 

Scedosporium apiospermum (Saccardo) Castellani and Chalmers 

Colonies are fast growing, greyish-white, suede-like to downy with a greyish-black reverse. 

Numerous single-celled, pale-brown, broadly clavate to ovoid conidia, 4-9 x 6- 

10 mm, rounded above with truncate bases are observed. Conidia are borne singly or 

in small groups on elongate, simple or branched conidiophores or laterally on hyphae. 

Conidial development can be described as annellidic, although the annellations (ringlike 

scars left at the apex of an annellide after conidial secession) are extremely difficult 

to see. Optimum temperature for growth is 30-37 O C. RG-2 organism. 

Ascocarp formation may be stimulated on cornmeal agar or other nutrient deficient 

media, however it should be noted that many isolates may fail to produce cleistothecia. 

Cleistothecia (non-ostiolate ascocarps) are yellow-brown to black, spherical, are mostly 

submerged in the agar and are composed of irregularly interwoven brown hyphae. 

When crushed cleistothecia release numerous, faintly brown, ellipsoidal ascospores, 

4-5 x 7- 9 µm in size. A Graphium synanamorph may also be present. 

The teleomorph is currently referred to as Pseudallescheria boydii, however as all 

species of Pseudallescheria have Scedosporium anamorphs, it is presumptive to use 

the teleomorph name to describe this fungus without seeing cleistothecia (ascocarps). 

Until the taxonomy of Pseudallescheria is resolved, it is recommended that laboratories 

use the anamorphic name Scedosporium apiospermum when describing this fungus. 

Scedosporium aurantiacum Gilgado et al. 

All isolates produce a light yellow diffusible pigment on PDA after a few days incubation. 

Conidiogenous cells and conidia are similar in shape and size to S. apiospermum, 

and the two can best be distinguished by genetic analysis. Conidiogenous cells arising 

from undifferentiated hyphae are cylindrical to slightly flask-shaped, producing slimy 

heads of one-celled , smooth-walled, subhyaline, obovoid or sub-cylindrical conidia. 

5-14 x 2-5 um. Erect synnemata (a Graphium synanamorph) may be present in some 

isolates, but the teleomorph is unknown. Optimum temperature for growth 37-40 O C, 

max 45 O C. RG-2 organism. 

S. apiospermum and S. aurantiacum appear to be common soil fungi capable of causing 

a spectrum of diseases similar in terms of variety and severity to those caused by 

Aspergillus collectively referred to as Pseudallescheriasis. The vast majority of infections 

are mycetomas, the remainder include infections of the eye, ear, central nervous 

system, internal organs and more commonly the lungs. 


(1980), Domsch et al. (1980), McGinnis et al. (1982), Campbell and Smith (1982), Rippon 

(1988), de Hoog et al. (2000) and Gilgado et al. (2005).

132 


Scedosporium apiospermum (Saccardo) Castellani and Chalmers 

Antifungal 

Culture and a cleistothecium of Pseudallescheria boydii. 

20 µm 

20 µm 

Conidiophores (annellides) and conidia of Scedosporium apiospermum. 

MIC µg/mL 

MIC µg/mL 

Range MIC90 Antifungal Range MIC90 Itraconazole 0.06-4 2 Amphotericin B 1-16 16 

Voriconazole 0.03-1 0.5 Posaconazole 0.5-2 1 

Good data available. McGinnis and Pasarell (1998), Espinel-Ingroff (2001, 2003), 

Espinel-Ingroff et al. (2001) and Cuenca-Estrella et al. (2006). MIC 90 s from the 

Australian Scedosporium Study.


Scedosporium aurantiacum Gilgado et al. 

Culture reverse (PDA) of S. apiospermum (left) and S. aurantiacum (right) showing 

the production of a light yellow diffusible pigment that is typical of S. aurantiacum. 

Antifungal 

20 µm 

Conidiophores (annellides) and conidia of Scedosporium aurantiacum. 

MIC µg/mL 

MIC µg/mL 

Range MIC90 Antifungal Range MIC90 Itraconazole 0.25-2 1 Amphotericin B 2-16 16 

Voriconazole 0.03-0.5 0.25 Posaconazole 0.125-1 0.5 

Good data available. Australian Scedosporium Study.

134 


Scedosporium prolificans (Hennebert & Desai) Gueho & de Hoog 

Synonym: Scedosporium inflatum Malloch & Salkin 

Colonies are rapid growing, flat, spreading, olive-grey to black and have a suede-like 

to downy surface texture. Conidia are borne in small groups on distinctive basally 

swollen, flask-shaped annellides, which occur singly or in clusters along the vegetative 

hyphae. Conidia are single-celled, hyaline to pale-brown, ovoid to pyriform, 2-5 x 3-13 

µm (average 3.4 x 5.3 µm), and have smooth thin walls. RG-2 organism. 

Key Features: dematiaceous hyphomycete with initial black pasty colony, basally 

swollen (inflated) annellides and no growth on media containing cycloheximide 

(actidione). 

For descriptions of species, keys to taxa and additional information see Malloch and 

Salkin (1984), Salkin et al. (1988), Rippon (1988), Wilson et al. (1990), Gueho and de 

Hoog (1991) and de Hoog et al. (2000). 

Antifungal 

20 µm 

Conidiophores (annellides) and conidia of Scedosporium prolificans. 

MIC µg/mL 

MIC µg/mL 

Range MIC90 Antifungal Range MIC90 Itraconazole 1-32 >8 Amphotericin B 1-16 >8 

Voriconazole 0.5-32 >8 Posaconazole >8 >8 

Good data available. McGinnis and Pasarell (1998), Espinel-Ingroff (2001, 2003), 

Espinel-Ingroff et al. (2001), Sabatelli et al. (2006), Cuenca-Estrella et al. (2006) and 

the Australian Scedosporium Study.


Schizophyllum commune Fries 

Colonies on 2% malt extract agar are spreading, woolly, whitish to pale greyish-brown, 

soon forming macroscopically visible fruiting bodies. Although some isolates may take 

up to 12 weeks to form fruiting bodies. Fruit bodies are sessile, kidney-shaped, lobed 

with split gills on the lower side. Hyphae are hyaline, wide and have clamp connections 

(although many primary clinical isolates are monokaryotic and will therefore not 

produce clamp connections). Basidia bear 4 basidiospores on erect sterigmata. Basidiospores 

hyaline, smooth-walled, elongate with lateral scar at lower end, 6-7 x 2-3 

µm. 

Shizophyllum is a common bracket fungus on rotten wood, and is an occasional human 

pathogen, principally associated with sinusitis, allergic bronchopulmonary mycosis 

and as a contaminant from respiratory specimens. RG-1 organism. 

Note: many clinical isolates of S. commune are monokaryotic and therefore do not 

show clamp connections, therefore any white, rapidly growing, sterile isolate showing 

good growth at 37 O C with tolerance to benomyl, susceptibility to cycloheximide, and a 

pronounced odour should be suspected of being S. commune (Sigler et al., 1995). 


(1980), Rippon (1988), Sigler et al. (1995) and de Hoog et al. (2000). 

Basidiocarps of Schizophyllum commune on malt extract agar.

136 


Scopulariopsis Bain 

Colonies are fast growing, varying in colour from white, cream, grey, buff to brown, 

black, but are predominantly light brown. Microscopic morphology shows chains of 

single-celled conidia produced in basipetal succession from by a specialised conidiogenous 

cell called an annellide. Once again, the term basocatenate can be used 

to describe such chains of conidia where the youngest conidium is at the basal end of 

the chain. In Scopulariopsis, annellides may be solitary, in groups, or organised into 

a distinct penicillus. Conidia are globose to pyriform, usually truncate, with a rounded 

distal portion, smooth to rough, and hyaline to brown in colour. 

Most members of the genus Scopulariopsis are soil fungi, however a few, in particular 

S. brevicaulis, have been reported as causative agents of onychomycosis and hyalohyphomycosis. 

RG-2 for species isolated from humans. 

Key Features: hyphomycete, conidia often shaped like light globes, basocatenate 

arising from annellides. 

For descriptions of species, keys to taxa and additional information see Morton and 

Smith (1963), Domsch et al. (1980), McGinnis (1980), Rippon (1988), Samson et al. 


Antifungal 

10 µm 

Conidiophores (annellides) and conidia of Scopulariopsis brevicaulis. 




Amphotericin B 2-16 Itraconazole 32 Voriconazole 2-8 




Sepedonium Link ex Greville 

Colonies are moderately fast growing, usually white to golden yellow, suede-like to 

downy, becoming fluffy with age. Conidiophores are hyaline and non-specialised, resembling 

short branches of the vegetative hyphae. Conidia are terminal, solitary, or 

in clusters, one-celled, globose to ovoid, 7 to 17 µm, hyaline to amber, smooth to verrucose 

and usually with a thick wall. 

The microscopic morphology of Sepedonium isolates resembles that of Histoplasma 

capsulatum. Therefore, positive identification of the latter, requires conversion of the 

mould form to the yeast phase by growth at 37 O C on enriched media or by exoantigen 

test. RG-1 organism. 

Key Features: hyphomycete, producing large, thick-walled, one-celled, verrucose, 

globose, terminal macroconidia from non-specialised conidiophores, resembling Histoplasma 

capsulatum. 


(1980) and Rippon (1988). 

Conidia of Sepedonium. 

15 µm

138 


Sporothrix schenckii Hektoen & Perkins 

Colonies (SDA) at 25 O C, are slow growing, moist and glabrous, with a wrinkled and 

folded surface. Some strains may produce short aerial hyphae and pigmentation may 

vary from white to cream to black. Conidiophores arise at right angles from thin septate 

hyphae and are usually solitary, erect and tapered toward the apex. Conidia are 

formed in clusters on tiny denticles by sympodial proliferation at the apex of the conidiophore, 

their arrangement often suggestive of a flower. As the culture ages, conidia 

are subsequently formed singly along the sides of both conidiophores and undifferentiated 

hyphae. Conidia are ovoid or elongated, 3-6 x 2-3 µm, hyaline, one-celled and 

smooth-walled. In some isolates, solitary, darkly-pigmented, thick-walled, one-celled, 

obovate to angular conidia may also be observed along the hyphae. 

On brain heart infusion agar containing blood at 37 O C, colonies are glabrous, white to 

greyish-yellow and yeast-like consisting of spherical or oval budding yeast cells. 

Sporothrix schenckii is a dimorphic fungus and has a world-wide distribution, particularly 

in tropical and temperate regions. It is commonly found in soil and on decaying 

vegetation and is a well known pathogen of humans and animals. Sporotrichosis is 

primarily a chronic mycotic infection of the cutaneous or subcutaneous tissues and adjacent 

lymphatics characterised by nodular lesions which may suppurate and ulcerate. 

Infections are caused by the traumatic implantation of the fungus into the skin, or very 

rarely, by inhalation into the lungs. Secondary spread to articular surfaces, bone and 

muscle is not infrequent, and the infection may also occasionally involve the central 

nervous system, lungs or genitourinary tract. RG-2 organism. 

Key Features: hyphomycete characterised by thermal dimorphism and clusters of 

ovoid, denticulate conidia produced sympodially on short conidiophores. 


(1980), Domsch et al. (1980), de Hoog et al. (1985), Rippon (1988) and de Hoog et al. 

(2000). 

10 µm 

Periodic Acid-Schiff (PAS) stained tissue section showing budding 

yeast-like cells of S. schenckii.


Antifungal 

Sporothrix schenckii Hektoen & Perkins 

10 µm 

Culture on SDA at 25 O C and budding yeast cells in BHI at 37 O C. 

10 µm 

Conidiophores and conidia of Sporothrix schenckii on SDA at 25 O C. 

MIC µg/mL 

MIC µg/mL 

Range MIC90 Antifungal Range MIC90 Fluconazole 32->64 >64 Amphotericin B 0.06->16 >16 

Itraconazole 0.03->16 0.5 (4) Flucytosine 4->64 >64 

Posaconazole 0.125-4 2 Caspofungin 1->8 nd 

Voriconazole 0.125->16 4 (16) Anidulafungin 0.25->8 nd 


McGinnis et al. (2001), Espinel-Ingroff et al. (2001), Espinel-Ingroff (2003), Gonzales 

et al. (2005) and Alvarado-Ramirez and Torres-Rodriguez (2007). (nd = not done).

140 


Stemphylium Wallroth 

Colonies are rapid growing, brown to olivaceous-black or greyish and suede-like to 

floccose. Microscopically, solitary, darkly pigmented, terminal, multicellular conidia 

(dictyoconidia) are formed on a distinctive conidiophore with a darker terminal swelling. 

Note: the conidiophore proliferates percurrently through the scar where the terminal 

conidium (poroconidium) was formed. Conidia are pale to mid-brown, oblong, 

rounded at the ends, ellipsoidal, obclavate or subspherical and are smooth or in part 

verrucose. Stemphylium should not be confused with Ulocladium which produces similar 

dictyoconidia from a sympodial conidiophore, not from a percurrent conidiogenous 

cell as in Stemphylium. RG-1 organism. 

Key Features: dematiaceous hyphomycete producing darkly pigmented, dictyoconidia 

from the swollen end of a percurrent conidiophore. 


1976), Rippon (1988) and de Hoog et al. (2000). 

20 µm 

Conidiophores and conidia of Stemphylium.


Syncephalastrum Schröter 

The genus Syncephalastrum is characterised by the formation of cylindrical merosporangia 

on a terminal swelling of the sporangiophore. Sporangiospores are arranged 

in a single row within the merosporangia. Syncephalastrum racemosum is the type 

species of the genus and a potential human pathogen; however, well-documented 

cases are lacking. It is found mainly from soil and dung in tropical and subtropical regions. 

It can also be a difficult laboratory aerial contaminant. The sporangiophore and 

merosporangia of Syncephalastrum species may also be mistaken for an Aspergillus 

species, if the isolate is not looked at carefully. 

Colonies are very fast growing, cottony to fluffy, white to light grey, becoming dark 

grey with the development of sporangia. Sporangiophores are erect, stolon-like, often 

producing adventitious rhizoids, and show sympodial branching (racemose branching) 

producing curved lateral branches. The main stalk and branches form terminal, globose 

to ovoid vesicles which bear finger-like merosporangia directly over their entire 

surface. At maturity, merosporangia are thin-walled, evanescent and contain 5-10(18) 

globose to ovoid, smooth-walled sporangiospores (merospores). Optimum growth 

temperature 20-40 O C. RG-2 organism. 

Key Features: zygomycete producing sympodially branching sporangiophores with 

terminal vesicles bearing merosporangia. 


(1980), McGinnis (1980), Onions et al. (1981), Rippon (1988), Samson et al. (1995), 

Hoog et al. (2000) and Ellis (2005b). 

30 µm 

10 µm 

Terminal vesicle, merosporangia and merospores of S. racemosum.

142 


Trichoderma Persoon ex Grey 

Colonies are fast growing, at first white and downy, later developing yellowish-green 

to deep green compact tufts, often only in small areas or in concentric ring-like zones 

on the agar surface. Conidiophores are repeatedly branched, irregularly verticillate, 

bearing clusters of divergent, often irregularly bent, flask-shaped phialides. Conidia 

are mostly green, sometimes hyaline, with smooth or rough walls and are formed in 

slimy conidial heads (gloiospora) clustered at the tips of the phialides. 

Trichoderma is a very common genus especially in soil and decaying wood. Gliocladium 

(with strongly convergent phialides) and Verticillium (with straight and moderately 

divergent phialides) are closely related genera. Trichoderma is an RG-1 organism. 

Key Features: hyphomycete with repeatedly branched conidiophores bearing clusters 

of divergent, flask-shaped phialides. 


(1980), McGinnis (1980), Rippon (1988), Samson et al. (1995), de Hoog et al. (2000). 

Antifungal 

20 µm 

Phialides and conidia of Trichoderma harzianum. 




Amphotericin B 0.5-2 Itraconazole 2-16 Voriconazole 0.25-2 


Espinel-Ingroff (2001) and WCH in-house data.


Trichophyton Malmsten 

The genus Trichophyton is characterised by the development of both smooth-walled 

macro- and microconidia. Macroconidia are mostly borne laterally directly on the hyphae 

or on short pedicels, and are thin- or thick-walled, clavate to fusiform, and range 

from 4-8 x 8-50 mm in size. Macroconidia are few or absent in many species. Microconidia 

are spherical, pyriform to clavate or of irregular shape and range from 2-3 x 2-4 

mm in size. The presence of microconidia distinguishes this genus from Epidermophyton 

and the smooth-walled, mostly sessile macroconidia separate it from Microsporum. 

Twenty species have been recognised, however only the more common species are 

included in these descriptions. 

In practice, two groups may be recognised on direct microscopy: 

1. Those species that usually produce microconidia, macroconidia may or may not be 

present i.e. T. rubrum, T. interdigitale, T. mentagrophytes, T. equinum, T. erinacei, 

T. tonsurans, T. terrestre and to a lesser extent T. verrucosum, which may produce 

conidia on some media; and 

2. Those species that usually do not produce conidia. Chlamydospores or other 

hyphal structures may be present, but microscopy is generally non-diagnostic; i.e. 

T. verrucosum, T. violaceum, T. concentricum, T. schoenleinii and T. soudanense. 

Many laboratories seem to have difficulty in distinguishing between species of Trichophyton, 

especially isolates of T. rubrum, T. interdigitale, T. mentagrophytes and T. 

tonsurans. Basically, the laboratories which consistently identify these fungi correctly 

do more work and use additional media and/or confirmatory tests. However, it must 

be stressed that no one single test is infallible, dermatophyte species are very variable 

organisms and many characteristics either overlap or are inconsistent. 

The Mycology Unit at the Adelaide Women’s and Children’s Hospital uses a dermatophyte 

identification scheme, devised by the late Geraldine Kaminski, comprising 6 

different media to help identify and differentiate the various species and strains of 

Trichophyton. The media in this scheme are Littman Oxgall agar, Lactritmel agar, 

Sabouraud’s agar with 5% NaCl, 1% Peptone agar, Trichophyton agar No. 1, and hydrolysis 

of urea (see appendix for details). 

Note: species concepts in dermatophytes are currently in a state of flux. Recent molecular 

studies have shown that many species appear to be clonal and that there is 

little correlation between “genetic” and “phenotypic” species (Graser et al. 2006). The 

descriptions and species concepts provided in this publication are based on traditional 

morphological criteria which may not correspond to molecular identification results. 

For description of species, keys to taxa and additional information see Rebell and 

Taplin (1970), Ajello (1972), Vanbreusegham et al. (1978), Rippon (1988), McGinnis 

(1980), Domsch et al. (1980), Kane et al. (1997) and de Hoog et al. (2000). 

Antifungal 

MIC µg/mL 

MIC µg/mL 

Range MIC90 Antifungal Range MIC90 Griseofulvin 0.06-4 1-2 Amphotericin B 0.03-16 0.5-1 

Itraconazole 0.01-8 0.25-0.5 Fluconazole 0.06->64 32 


Fernandez-Torres et al. (2001), Sabatelli et al. (2006), Santos and Hamdan (2006).

144 


Trichophyton ajelloi (Vanbreuseghem) Ajello 

Teleomorph: Arthroderma uncinatum Dawson and Gentles 

Colonies are usually flat, powdery, cream to tan to orange-tan in colour, with a blackish-purple 

submerged fringe and reverse. Macroconidia are numerous, smooth, thickwalled, 

elongate, cigar-shaped, 29-65 x 5-10 µm, and multiseptate with up to 9 or 10 

septa. Microconidia are usually absent, but when present are ovate to pyriform in 

shape. RG-1 organism. 

Trichophyton ajelloi is a geophilic fungus with a world-wide distribution which may occur 

as a saprophytic contaminant on humans and animals. Infections in man and animals 

doubtful. Not known to invade hair in vivo, but produces hair perforations in vitro. 

Key Features: culture characteristics, macroconidial morphology, urease positive and 

good growth on Sabouraud’s 5% salt agar. 

20 µm 

Culture and macroconidia of Trichophyton ajelloi.


Trichophyton concentricum Blanchard 

Teleomorph: Arthroderma benhamiae Ajello & Chang. 

Colonies (SDA) are slow growing, raised and folded, glabrous becoming suede-like, 

mostly white to cream-coloured, but sometimes orange-brown-coloured, often deeply 

folded into the agar which may produce splitting of the medium in some cultures. Reverse 

is buff to yellow-brown to brown in colour. Cultures consist of broad, muchbranched, 

irregular, often segmented, septate hyphae which may have “antler” tips 

resembling T. schoenleinii. Chlamydospores are often present in older cultures. Microconidia 

and macroconidia are not usually produced, although some isolates will 

produce occasional clavate to pyriform microconidia. Note: hyphal segments may 

artificially resemble macroconidia. RG-2 organism. 

Kaminski’s Dermatophyte Identification Scheme 

Littman Oxgall Agar: Small, heaped and folded, blue-grey, suede to glabrous colonies 

with no reverse or diffusible pigment. 

Lactritmel Agar: Colonies tend to be flatter, dull-white, suede-like, almost glabrous 

with a yellow-brown reverse pigment. Microscopic morphology is similar to that described 

above, however occasional clavate to pyriform microconidia may be present. 

Note: occasional, small, slender, smooth thin-walled macroconidia with 2-3 septa have 

been reported on specialised media like lactritmel agar, bean pod agar and hay infusion 

agar. 

Sabouraud’s Dextrose Agar with 5% NaCl: Small, stunted, heaped and folded, 

cream to buff, glabrous colonies with no reverse pigment. 

1% Peptone Agar: Flat, white, suede-like colony with a pale yellow-brown reverse. 

Hydrolysis of Urea: Negative after 7 days 

Vitamin Free Agar (Trichophyton Agar No.1): Growth occurs on vitamin free agar 

(T1) but is usually slightly better on media containing thiamine i.e. T3 = T1 + thiamine 

and inositol, and T4 = T1 + thiamine. The slight enhancement of growth in the presence 

of thiamine helps to distinguish T. concentricum from T. schoenleinii, although 

this does not occur in all strains. 

Hair Perforation Test: Negative at 28 days. 

Trichophyton concentricum is an anthropophilic fungus which causes chronic widespread 

non-inflammatory tinea corporis known as tinea imbricata because of the concentric 

rings of scaling it produces. It is not known to invade hair. Infections among 

Europeans are rare. Distribution is restricted to the Pacific Islands of Oceania, South 

East Asia and Central and South America. 

Key Features: hyphomycete (dermatophyte), clinical disease, geographical distribution 

and culture characteristics.

146 


Trichophyton concentricum Blanchard 

20 µm 

T. concentricum on mycobiotic agar showing a typical slow 

growing, heaped and folded, glabrous to suede like colony. 

Microscopic morphology of T. concentricum showing the formation 

of typical “balloon-shaped” chlamydospores. Note: microconidia 

and macroconidia are usually not produced.


Trichophyton equinum (Matruchot & Dassonville) Gedoelst 

Teleomorph: Arthroderma vanbreuseghemii Takashio 

Colonies (SDA) are usually flat, but some may develop gentle folds or radial grooves, 

white to buff in colour, suede-like to downy in texture, and are similar to T. mentagrophytes. 

Cultures usually have a deep-yellow submerged fringe and reverse which 

later becomes dark red in the centre. Microscopically, abundant microconidia which 

may be clavate to pyriform and sessile or spherical and stalked are formed laterally 

along the hyphae. Macroconidia are only rarely produced, but when present are clavate, 

smooth, thin-walled and of variable size. Occasional nodular organs may be 

present and the microconidia often undergo a transformation to produce abundant 

chlamydospores in old cultures. RG-2 organism. 


Littman Oxgall Agar: Compact, raised velvety white surface with yellowish reverse 

and diffusing yellow pigment. 

Lactritmel Agar: Flat spreading, white to cream-coloured, powdery to granular surface 

with a central downy papilla, and deep brownish red reverse. Microscopic morphology 

as described above for the primary culture. 

Sabouraud’s Dextrose Agar with 5% NaCl: An extremely stunted, slow growing dark 

brown colony with a few submerged brown hyphae. 

1% Peptone Agar: Flat white to cream, suede-like surface with raised white downy 

centre and pale yellow reverse. 

Hydrolysis of Urea: Positive in 4-5 days. 

Nutritional Tests on Trichophyton Agars: Most strains require nicotinic acid (Fig. 1) 

for growth except those from Australia and New Zealand, which are autotrophic (Fig. 

2). T1 = vitamin free agar, T5 = vitamin free + nicotinic acid agar. 

Fig. 1. Fig. 2. 

Hair Perforation Test: Negative; but positive for the autotrophicum strains. 

Key Features: microscopic morphology, culture characteristics, nicotinic acid requirement 

and clinical lesions in horses.

148 


Trichophyton equinum (Matruchot & Dassonville) Gedoelst 

Trichophyton equinum is a zoophilic fungus causing ringworm in horses and rare infections 

in humans. It has a world-wide distribution except for the autotrophicum strain 

which is restricted to Australia and New Zealand. Invaded hairs show an ectothrix 

infection but do not fluoresce under Wood’s ultra-violet light. 

20 µm 

Culture, microconidia, macroconidia and nodular organs of T. equinum.


Trichophyton erinacei (Smith & Marples) Quaife 


Colonies (SDA) are white, flat, powdery, sometimes downy to fluffy with a brilliant lemon 

yellow reverse. Numerous large clavate microconidia are borne on the sides of 

hyphae. Macroconidia are smooth-walled, two- to six-celled, clavate, variable in size, 

and may have terminal appendages. Macroconidia are much shorter than those seen 

in T. mentagrophytes. RG-2 organism. 


Littman Oxgall Agar: White, downy colony with yellowish-green diffusible pigment. 

Lactritmel Agar: White suede-like to powdery colony with brilliant yellow reverse. 

Numerous large slender clavate microconidia. 

Sabouraud’s Dextrose Agar with 5% NaCl: White folded suede-like to powdery 

colony with no reverse pigment. 

1% Peptone Agar: White, suede-like to powdery colony with pale yellow reverse. 

Hydrolysis of Urea: Negative at 7 days. 


nutritional requirements. Colonies are white suede-like to powdery with no reverse 

pigment. 

Hair Perforation Test: Positive 

Trichophyton erinacei is a zoophilic fungus associated with hedgehogs and the epidermal 

mites which they often harbour. Human infections occur most frequently on the 

exposed parts of the body, but tinea of the scalp and nails can also occur. Invaded 

hairs show an ectothrix infection but do not fluoresce under Wood’s ultra-violet light. 

The distribution of this fungus is New Zealand and Europe. 

Positive “in vitro” hair perforation test.

150 


Trichophyton erinacei (Smith & Marples) Quaife 

Key Features: culture characteristics, microscopic morphology, geographical 

distributions and negative urease test. 

20 µm 

Culture, microconidia and macroconidia of Trichophyton erinacei. 

Trichophyton erinacei is generally distinguished from T. mentagrophytes by (a) its 

microscopic morphology showing numerous large slender clavate microconidia borne 

on the sides of hyphae and its smooth, thin-walled clavate macroconidia; (b) its brilliant 

lemon yellow reverse pigment on plain Sabouraud’s agar and Lactritmel agar; (c) its 

lack of reverse pigment on Sabouraud’s salt agar; and (d) its negative hydrolysis of 

urea.


Teleomorph: Arthroderma vanbreuseghemii Takashio 

Colonies (SDA) are usually flat, white to cream in colour with a powdery to suede-like 

surface and yellowish and pinkish brown reverse pigment, often becoming a darker 

red-brown with age. Numerous subspherical to pyriform microconidia, occasional spiral 

hyphae and spherical chlamydospores are present, the latter being more abundant 

in older cultures. Occasional slender, clavate, smooth-walled, multiseptate macroconidia 

are also present in some cultures. RG-2 organism. 


Littman Oxgall Agar: raised white downy colony with no reverse pigment. 

Lactritmel Agar: Macroscopic and microscopic features as described for the primary 

culture. 

Sabouraud’s Dextrose Agar with 5% NaCl: Heaped and folded, buff-coloured suedelike 

surface with a dark reddish-brown submerged fringe and brown reverse. 

1% Peptone Agar: Flat, white to cream, suede-like surface with raised white downy 

centre. No reverse pigment. 

Hydrolysis of Urea: Positive within 5 days. 


nutritional requirements, flat cream powdery surface with central downy tuft. Reverse 

pale pinkish-brown. 


Trichophyton interdigitale Priestley 

Key Features: culture characteristics, microscopic morphology and in vitro perforation 

of human hair. 

Trichophyton interdigitale can be distinguished from T. rubrum and T. mentagrophytes 

by (a) its culture characteristics and microscopic morphology on Sabouraud’s dextrose 

agar and/or lactritmel agar; (b) its growth and colony morphology on Sabouraud’s salt 

agar (colonies of T. interdigitale and T. mentagrophytes unlike T. rubrum, grow very 

well on this medium and usually produce a distinctive dark reddish-brown reverse pigment); 

(c) a positive urease test (within 7 days), a positive hair perforation test and the 

production of a yellow-brown to pinkish-brown reverse pigment on pigment stimulation 

media like lactritmel and Trichophyton No.1 agars; and (d) on 1% peptone agar 

T. interdigitale has a suede-like to downy surface whereas T. mentagrophytes has a 

characteristic granular appearance. 

Trichophyton interdigitale is an anthropophilic fungus which is a common cause of 

tinea pedis, particularly the vesicular type, tinea corporis, and sometimes superficial 

nail plate invasion. It is not known to invade hair in vivo but produces hair perforations 

in vitro. Distribution is world-wide.

152 


20 µm 

Trichophyton interdigitale Priestley 

20 µm 

Culture, microconidia, macroconidia, chlamydospores and spiral hyphae in 

Trichophyton interdigitale.


Trichophyton interdigitale var. nodulare 

Supplementary description for Trichophyton interdigitale var. nodulare, a dysgonic variant 

of T. interdigitale with distinctive bright yellow to apricot-coloured colonies with a 

suede-like to powdery surface and a bright yellow-brown to orange reverse. On primary 

isolation, colonies are often glabrous with minimal surface mycelium. Microscopically 

characteristic “nodular organs” are observed in the vegetative hyphae. Usually, 

no conidia are seen but some isolates, especially with subculture, may produce subspherical 

to pyriform microconidia similar to those of T. interdigitale. RG-2 organism. 

T. interdigitale var. nodulare is an unusual cause of tinea pedis. It has a world-wide distribution. 

It is not known to invade hair in vivo, but produces hair perforations in vitro. 

20 µm 

Culture and “nodular organs” T. interdigitale var. nodulare.

154 


Trichophyton mentagrophytes (Robin) Blanchard 

Teleomorph: Arthroderma simii Stockdale et al. 

Colonies (SDA) are generally flat, white to cream in colour, with a powdery to granular 

surface. Some cultures show central folding or develop raised central tufts or pleomorphic 

suede-like to downy areas. Reverse pigmentation is usually a yellow-brown to 

reddish-brown colour. Numerous single-celled microconidia are formed, often in dense 

clusters. Microconidia are hyaline, smooth-walled, and are predominantly spherical to 

subspherical in shape, however occasional clavate to pyriform forms may occur. Varying 

numbers of spherical chlamydospores, spiral hyphae and smooth, thin-walled, clavate-shaped, 

multi-celled macroconidia may also be present. RG-2 organism. 


Littman Oxgall Agar: Raised greyish-white, suede-like to downy colony. Some cultures 

may show a diffusible yellow to brown pigment. 

Lactritmel Agar: Cultures are flat, white to cream in colour, with a powdery to granular 

surface. Some cultures develop a raised central tuft or pleomorphic downy areas. Reverse 

pigmentation is yellow-brown to pinkish-brown to red-brown. Microscopic morphology 

similar to that described above, with predominantly spherical microconidia, 

often forming in dense clusters and varying numbers of spherical chlamydospores, 

spiral hyphae and smooth, thin-walled, clavate, multiseptate macroconidia. 

Sabouraud’s Dextrose Agar with 5% NaCl: Cultures are heaped and folded, buff to 

brown in colour, with a suede-like surface texture and characteristically have a very 

dark reddish-brown submerged peripheral fringe and reverse pigmentation. 

1% Peptone Agar: Flat, cream-coloured, powdery to granular colony with no reverse 

pigment. 

Hydrolysis of Urea: Positive within 7 days (usually 3 to 5 days). 


nutritional requirements. Cultures are flat, cream-coloured, with a powdery to suedelike 

surface, and have a reddish-brown reverse pigmentation. 

Hair Perforation Test: Positive within 14 days. 

Key Features: culture characteristics, microscopic morphology and clinical disease 

with known animal contacts. T. mentagrophytes can be distinguished from T. interdigitale 

by (a) its granular appearance on 1% Peptone agar, (b) its microscopic morphology 

of more spherical microconidia and generally greater numbers of macroconidia 

and (c) a yellow to brown diffusible pigment is often seen on Littman Oxgall agar. 

T. mentagrophytes is the zoophilic form of T. mentagrophytes with a world-wide distribution 

and a wide range of animal hosts including mice, guinea-pigs, kangaroos, cats, 

horses, sheep and rabbits. Produces inflammatory skin or scalp lesions in humans, 

particularly in rural workers. Kerion of the scalp and beard may occur. Invaded hairs 

show an ectothrix infection but do not fluoresce under Wood’s ultra-violet light.


Trichophyton mentagrophytes (Robin) Blanchard 

20 µm 20 µm 

20 µm 

Cultures, microconidia, macroconidia, chlamydospores and spiral hyphae in 

Trichophyton mentagrophytes.

156 


Trichophyton mentagrophytes var. quinckeanum (Zopf) MacLeod & Muende 

Colonies (SDA) are flat or slightly raised and folded, white to cream, suede-like in 

texture with a pale yellow-brown to pinkish brown reverse. A characteristic pungent 

“mousy” odour may be present. Numerous microconidia, predominantly slender clavate 

when young, are borne laterally along the sides of hyphae. With age the microconidia 

become broader and pyriform, and some subspherical forms are present. Occasional 

to moderate numbers of smooth-walled, multiseptate, clavate macroconidia 

may be present. RG-2 organism. 


Littman Oxgall Agar: Raised, dome-like bluish-grey, suede-like colony with a narrow 

flat, greyish-white, suede-like border. No diffusible or reverse pigment should be 

present. 

Lactritmel Agar: Flat, white to cream, suede-like to powdery colony with either no 

reverse pigment or a pale yellow-brown to pinkish-brown reverse. Numerous slender 

clavate to pyriform (depending on age of sub-culture) microconidia and occasional to 

moderate numbers of smooth, thin-walled, clavate macroconidia are present. 

Sabouraud’s Dextrose Agar with 5% NaCl: Heaped up and much folded white 

suede-like colony with very pale yellow-brown reverse. No submerged fringe. 

1% Peptone Agar: Raised white suede-like to downy colony with no reverse 

pigment. 

Hydrolysis of Urea: Positive within 7 days (usually very rapid 2-3 days). 

Vitamin Free Agar (Trichophyton Agar No.1): Flat, white to cream, suede-like 

colony with either no reverse pigment or a pale yellow-brown reverse. i.e. no special 

nutritional requirements. 

Hair Perforation Test: Positive in 7 to 10 days. 

Key Features: culture characteristics, microscopic morphology, contact with mice, 

odour and rapid urease test. 

T. mentagrophytes var. quinckeanum may be distinguished from T. mentagrophytes by 

(a) its characteristic culture appearance on Littman Oxgall agar (i.e. raised, dome-like, 

bluish-grey suede-like colony with a narrow flat, greyish-white, suede-like border and 

no diffusible or reverse pigment); and on Sabouraud’s salt agar (typically heaped and 

folded white suede-like colony, but with no distinctive dark reddish-brown submerged 

fringe and reverse pigment as seen in T. mentagrophytes); (b) microscopic morphology 

showing numerous slender clavate with some pyriform microconidia and moderate 

numbers of smooth thin-walled, clavate macroconidia; (c) a rapid urease test, usually 

within 2 to 3 days; and (d) cultures often have a characteristic pungent “mousy” 

odour.


Trichophyton mentagrophytes var. quinckeanum (Zopf) MacLeod & Muende 

Trichophyton mentagrophytes var. quinckeanum causes “mouse favus” on mice, and 

this is seen as thick, yellow, saucer-shaped crusted lesions up to 1 cm in diameter 

called scutula. Invaded hairs are rarely seen but they may show either ectothrix or 

endothrix infection. Infected human hairs do not fluoresce under Wood’s ultra-violet 

light, but very occasional hairs from experimental lesions in guinea pigs may show a 

pale yellow fluorescence. The geographical distribution of this dermatophyte is difficult 

to establish, but it is probably world-wide. It is often associated with mice plagues in 

the Australian Wheat Belt. 

20 µm 

Culture, microconidia and macroconidia of T. mentagrophytes var. 

quinckeanum.

158 


Trichophyton rubrum (Castellani) Semon 

Many strains and varieties of T. rubrum have been described and opinion differs between 

mycologists as to the exact validity of many of these. For practical purposes we 

will distinguish two types: T. rubrum downy type and T. rubrum granular type. 

Microscopically, the downy type is characterised by the production of scanty to moderate 

numbers of slender clavate microconidia and no macroconidia. It frequently causes 

chronic infections of skin and nails. Granulomatous lesions may sometimes occur. 

Microscopically, the granular type is characterised by the production of moderate to 

abundant numbers of clavate to pyriform microconidia and moderate to abundant numbers 

of thin-walled, cigar-shaped macroconidia. The macroconidia may or may not 

have terminal appendages. 

Trichophyton rubrum granular strain is a frequent cause of tinea corporis in South East 

Asia and in Aborigines living in the Northern Territory of Australia. However, since the 

Vietnam War, it has been spread throughout the world, especially to those countries 

with returning troops or to those receiving refugees, where it has often been described 

as a new species. The granular strain of T. rubrum represents the parent strain of T. 

rubrum downy type; the latter evolved by establishing a niche in feet (tinea pedis) when 

the former was imported into Europe and North America at the beginning of the 20th 

century. It should be stressed that intermediate strains between the two types do occur 

and that many culture and morphological characteristics overlap. 

Trichophyton rubrum typical downy strain 

Colonies (SDA) are flat to slightly raised, white to cream, suede-like to downy, with a 

yellow-brown to wine-red reverse. Most cultures show scanty to moderate numbers of 

slender clavate to pyriform microconidia. Macroconidia are usually absent, however 

closterospore-like projections may be present in some mounts. Note: on primary isolation 

some cultures may lack reverse pigmentation and fail to produce microconidia. 

These will need to be subcultured onto media like Lactritmel agar or potato dextrose 

agar which stimulate pigmentation and sporulation. If sporulation still fails subculture 

the fungus onto Trichophyton Agar No.1. RG-2 organism. 

Culture of Trichophyton rubrum downy strain.


Trichophyton rubrum (Castellani) Semon 


Littman Oxgall Agar: Raised, greyish-white, suede-like to downy colony with no reverse 

pigment. Some cultures may have showed a faint greenish-yellow diffusible 

pigment. 

Lactritmel Agar: Flat, white, downy colonies with a deep wine-red reverse pigment. 

Microscopically, cultures show the typical downy type morphology of pyriform to slender 

clavate microconidia. 

Sabouraud’s Dextrose Agar with 5% NaCl: Very stunted, white downy colony with a 

pale yellow-brown reverse pigment. 

1% Peptone Agar: Flat, white to cream, downy colony often with a raised centre. No 

reverse pigment produced. 

Hydrolysis of Urea: Negative at 7 days. 


vitamin requirements. Colonies are flat, white to cream, suede-like to downy with a 

deep wine-red reverse pigment. 

Hair Perforation Test: Negative at 28 days. 

Key Features: culture characteristics, microscopic morphology and failure to perforate 

hair “in vitro”. 

20 µm 

Typical slender clavate microconidia of Trichophyton rubrum downy type.

160 


Colonies (SDA) are flat to slightly raised, white to cream, suede-like with a pinkish-red 

reverse. Microscopically, most cultures have numerous clavate to pyriform microconidia 

and moderate numbers of smooth, thin-walled multiseptate, slender cylindrical 

macroconidia. Older cultures may show numerous chlamydospores with few clavate 

to pyriform microconidia. 


Littman Oxgall Agar: Raised, greyish, suede-like colonies with some radial folding 

and a greenish-yellow diffusible pigment. 

Lactritmel Agar: Flat, white to rose pink, suede-like to granular colonies with a pinkish 

to wine-red reverse. Numerous broad clavate to pyriform microconidia and moderate 

numbers of smooth, thin-walled, slender cylindrical macroconidia are present. A few 

chlamydospores may be present in older cultures. 

Sabouraud’s Dextrose Agar with 5% NaCl: A very stunted, heaped and folded, 

glabrous, cream thallus, later developing a dark red central spot. Reverse is a brownishyellow 

colour. 

1% Peptone Agar: Flat, white to cream, glabrous to suede-like colony with no reverse 

pigment. 

Hydrolysis of Urea: Positive at 7 days 


nutritional requirements. White to cream suede-like colonies with a pinkish-red to 

wine-red reverse. 


Trichophyton rubrum granular type 

Key Features: culture characteristics and microscopic morphology. 

Culture of Trichophyton rubrum granular strain.


20 µm 

Trichophyton rubrum granular type 

20 µm 

Macroconidia and microconidia of Trichophyton rubrum granular type.

162 


Trichophyton schoenleinii (Lebert) Langeron & Milochevitch 

Colonies (SDA) are slow growing, waxy or suede-like with a deeply folded honeycomb-like 

thallus and some sub-surface growth. The thallus is cream-coloured to yellow 

to orange brown. Cultures are difficult to maintain in their typical convoluted form, 

and rapidly become flat and downy. No reverse pigmentation is present. No macroconidia 

and microconidia are seen in routine cultures, however numerous chlamydospores 

may be present in older cultures. However, characteristic antler “nail head” 

hyphae also known as “favic chandeliers” may be observed. A few distorted clavate 

microconidia may be formed by some isolates when grown on polished rice grains. 

RG-2 organism 

Key Features: clinical history, culture characteristics and microscopic morphology 

showing favic chandeliers. 

20 µm 

Culture and “favic chandeliers” of Trichophyton schoenleinii. 

Trichophyton schoenleinii is an anthropophilic fungus causing favus in humans. Favus 

is a chronic, scarring form of tinea capitis characterised by saucer-shaped crusted lesions 

or scutula and permanent hair loss. Invaded hairs remain intact and fluoresce 

a pale greenish yellow under Wood’s ultra-violet light. Favus was once common in 

Eurasia and North Africa, however its incidence is now in decline.


Trichophyton soudanense Joyeux. 

Colonies (SDA) are slow-growing with a flat to folded, suede-like surface. Often there 

is a broad fringe of submerged growth. The surface mycelium and reverse pigment are 

characteristically a deep apricot-orange in colour. Microscopically, the hyphae often 

show reflexive or right-angle branching. Pyriform microconidia may occasionally be 

present and numerous chlamydospores are often found in older cultures. On BCPmilk 

solids agar, a thin halo of clearing usually appears in the milk solids around the 

colony edge at 7-10 days. RG-2 organism. 

T. soudanense appears to be genetically related to Trichophyton rubrum and T. violaceum 

(Graser et al. 2007), however we have maintained the current description until 

the taxonomy is clarified. 

Key Features: clinical history, culture characteristics and microscopic morphology 

showing reflexive hyphal branching and endothrix invasion of hair. 

20 µm 

Culture and “reflexive” hyphal branching in Trichophyton soudanense. 

Trichophyton soudanense is an anthropophilic fungus which is a frequent cause of 

tinea capitis in Africa. Invaded hairs show an endothrix infection but do not fluoresce 

under Wood’s ultra-violet light. Distribution is mainly in Africa with imported cases now 

reported from Europe, Brazil, Australia and USA.

164 


Trichophyton terrestre Durie and Frey 

Teleomorphs: Arthroderma insingulare Padhye and Carmichael 

Arthroderma lenticulare Pore, Tsao and Plunkett 

Arthroderma quadrifidum Dawson and Gentles 

Colonies (SDA) are usually flat to downy with a suede-like to granular texture resembling 

T. mentagrophytes. The surface colour may range from white to cream, buff to 

yellow, or greenish-yellow. Reverse pigmentation is usually yellowish-brown although 

some variants have a deep rose-red reverse. Microconidia are large, clavate or pedicellate, 

usually exhibiting transition forms to more or less abundant lateral macroconidia. 

Macroconidia are clavate to cylindrical with rounded ends, smooth and thin-walled, 

and are two- to six-celled. Chlamydospores, hyphal spirals, racquet mycelium and 

antler hyphae may also be present. No growth at 37 O C. RG-1 organism. 

Trichophyton terrestre is a geophilic fungus of world-wide distribution which may occur 

as a saprophytic contaminant on humans and animals. It is not known to invade hair 

in vivo, but produces hair perforations in vitro. 

Key Features: culture characteristics and microscopic morphology. 

20 µm 

Culture and macroconidia of Trichophyton terrestre.


Colonies (SDA) show considerable variation in texture and colour. They may be suedelike 

to powdery, flat with a raised centre or folded, often with radial grooves. The colour 

may vary from pale-buff to yellow, (the sulfureum form which resembles Epidermophyton 

floccosum), to dark-brown. The reverse colour varies from yellow-brown to reddish-brown 

to deep mahogany. Hyphae are relatively broad, irregular, much branched 

with numerous septa. Numerous characteristic microconidia varying in size and shape 

from long clavate to broad pyriform, are borne at right angles to the hyphae, which often 

remain unstained by lactophenol cotton blue. Very occasional smooth, thin-walled, 

irregular, clavate macroconidia may be present on some cultures. Numerous swollen 

giant forms of microconidia and chlamydospores are produced in older cultures. RG-2 

organism. 


Littman Oxgall Agar: Restricted colony with cream, sometimes greyish, suede-like 

folded surface with no reverse pigment. 

Lactritmel Agar: Macroscopic and microscopic features as described above for the 

primary culture. 

Sabouraud’s Dextrose Agar with 5% NaCl: Very stunted slow growing colony with 

dark brown surface and reverse. 

1% Peptone Agar: Flat, white to cream suede-like surface with raised centre. No 

reverse pigment. 

Hydrolysis of Urea: positive at 5 days 

Trichophyton tonsurans Malmsten 

Nutritional Tests on Trichophyton Agars: results demonstrate a partial requirement 

for thiamine. T1 = vitamin free agar, T4 = vitamin free + thiamine agar. 

Hair Perforation Test: Positive within 14 days. 

T1 T4 

Key Features: microscopic morphology, culture characteristics, endothrix invasion of 

hairs and partial thiamine requirement. 

Trichophyton tonsurans is an anthropophilic fungus with a world wide distribution which 

causes inflammatory or chronic non-inflammatory finely scaling lesions of skin, nails 

and scalp. It is a common cause of tinea capitis in the Australian Aborigine and African 

Americans. Invaded hairs show an endothrix infection and do not fluoresce under 

Wood’s ultra-violet light.

166 


Trichophyton tonsurans Malmsten 

20 µm 

Colonies, hyphae, microconidia and macroconidia of Trichophyton tonsurans.



Colonies (SDA) are slow growing, small, button or disc-shaped, white to cream-coloured, 

with a suede-like to velvety surface, a raised centre, and flat periphery with 

some submerged growth. Reverse pigment may vary from non-pigmented to yellow. 

Broad, irregular hyphae with many terminal and intercalary chlamydospores are 

present. Chlamydospores are often in chains. The tips of some hyphae are broad 

and club-shaped, and occasionally divided, giving the so-called “antler” effect. When 

grown on thiamine-enriched media, occasional strains produce clavate to pyriform microconidia 

borne singly along the hyphae. Macroconidia are only rarely produced, but 

when present have a characteristic tail or string-bean shape. RG-2 organism. 

Confirmatory Tests: 

Trichophyton verrucosum Bodin 

Growth at 37 O C: unlike other dermatophytes growth is enhanced at 37 O C 

Nutritional Requirements: all strains require thiamine and approximately 80% require 

thiamine and inositol. There is no growth on casein vitamin free agar (T1), minimal 

submerged growth on T1 + inositol (T2), good growth on T1 + inositol and thiamine 

(T3) and good growth on T1 + thiamine only (T4). 

All strains produce typical chains of chlamydospores, often referred to as “chains of 

pearls”, especially when grown on BCP milk solids glucose agar at 37 O C. Also when 

grown at 25 O C on milk solids glucose agar a halo of peripheral clearing of milk solids 

occurs within 7 days. 

Microscopic examination of young 4 to 5 day old colonies, grown from a very small 

inoculum, on Sabouraud’s’ dextrose agar containing 0.5% yeast extract and incubated 

at 30 O C, show characteristic terminal vesicles (not chlamydospores) at the tips of 

hyphae. The number of vesicles produced is greater from primary inoculations of skin 

scrapings or hairs. 

Key Features: culture characteristics and requirements for thiamine and inositol, large 

ectothrix invasion of hair, clinical lesions and history. 

Trichophyton verrucosum is a zoophilic fungus causing ringworm in cattle. Infections 

in humans result from direct contact with cattle or infected fomites and are usually 

highly inflammatory involving the scalp, beard or exposed areas of the body (ie. nails, 

skin). Invaded hairs show an ectothrix infection and fluorescence under Wood’s ultraviolet 

light has been noted in cattle but not in humans. Geographic distribution is 

world-wide.

168 


Trichophyton verrucosum Bodin 

20 µm 

20 µm 

30 µm 

20 µm 

Trichophyton verrucosum showing clavate to pyriform microconidia, 

characteristic rat tail or string bean-shaped macroconidia, terminal vesicles 

at the tips of hyphae in young colonies and chains of chlamydospores.


Trichophyton violaceum Sabouraud apud Bodin 

Colonies (SDA) are very slow growing, glabrous or waxy, heaped and folded and a 

deep violet in colour. Cultures often become pleomorphic, forming white sectors and 

occasional non-pigmented strains may occur. Hyphae are relatively broad, tortuous, 

much branched and distorted. Young hyphae usually stain well in lactophenol cotton 

blue, whereas older hyphae stain poorly and show small central fat globules and granules. 

No conidia are usually seen, although occasional pyriform microconidia have 

been observed on enriched media. Numerous chlamydospores are usually present, 

especially in older cultures. RG-2 organism. 

Nutritional Requirements: T. violaceum has a partial nutrient requirement for thiamine. 

There is minimal growth on casein vitamin-free agar (Trichophyton Agar No. 1), 

and slightly better growth on vitamin-free agar plus thiamine (Trichophyton Agar No. 

4). The partial requirement for thiamine separates this organism from T. gourvillii, T. 

rubrum, and other species that may produce purple pigmented colonies. 

Key Features: culture characteristics, partial thiamine requirement and endothrix hair 

invasion. 

Culture and chlamydospores of Trichophyton violaceum. 

20 µm 

Trichophyton violaceum is an anthropophilic fungus causing inflammatory or chronic 

non-inflammatory finely scaling lesions of skin, nails, beard and scalp, producing the 

so-called “black dot” tinea capitis. Distribution is world-wide, particularly in the Near 

East, Eastern Europe, USSR and North Africa. Invaded hairs show an endothrix 

infection and do not fluoresce under Wood’s ultra-violet light.

170 


Trichosporon Behrend 

The genus Trichosporon is characterised by the development of hyaline, septate hyphae 

that fragment into oval or rectangular arthroconidia. Some blastoconidia are also 

seen. The colonies are usually raised and have a waxy appearance, which develop 

radial furrows and irregular folds. 

Following recent molecular studies, the genus has undergone major revision (Gueho 

et al. 1992, de Hoog et al. 2000, Rodriguez-Tudela et al. 2005) and 6 species of medical 

importance are described below. In particular, the name Trichosporon beigelii is 

now obsolete, and previously described infections reported in the literature under this 

name could in fact be due to any one of the species listed below. 

Trichosporon species are a minor component of normal skin flora, and are widely distributed 

in nature. They are regularly associated with the soft nodules of white piedra, 

and have been involved in a variety of opportunistic infections in the immunosuppressed 

patient. Disseminated infections are most frequently caused by T. asahii 

and have been associated with leukaemia, organ transplantation, multiple myeloma, 

aplastic anaemia, lymphoma, solid tumours and AIDS. Disseminated infections are 

often fulminate and widespread, with lesions occurring in the liver, spleen, lungs and 

gastrointestinal tract. Infections in non-immunosuppressed patients include endophthalmitis 

after surgical extraction of cataracts, endocarditis usually following insertion 

of prosthetic cardiac valves, peritonitis in patients on continuous ambulatory peritoneal 

dialysis (CAPD), and intravenous drug abuse. 

For descriptions of species, keys to taxa and additional information see Kurtzman 

and Fell (1988), Gueho et al. (1992), de Hoog et al. (2000), Rodriguez-Tudela et al. 

(2005). 

Key to medically important species (de Hoog et al. 2000). 

1. Growth with melibiose 2 

No growth with melibiose 3 

2. Tolerant to cycloheximide T. mucoides 

Not tolerant to cycloheximide T. cutaneum 

3. Growth with myo-inositol, no growth with L-arabinose T. inkin 

No growth with myo-inositol, growth with L-arabinose 4 

4. Colony with very slow growth; thallus consisting of clumps 

of meristematic cells (sarcinae) T. asteroides 

Colonies and microscopy otherwise 5 

5. Appressoria present in slide cultures T. ovoides 

Appressoria absent in slide cultures 6 

6. Arthroconidia barrel-shaped; thallus not meristematic T. asahii 

Arthroconidia elongate, or thallus meristematic T. asteroides


Trichosporon asahii Akagi ex Sugita et al. 

Colonies (SDA) are white to cream-coloured, powdery, suede-like to farinose with radial 

furrows and irregular folds. Budding cells and lateral conidia are absent. Arthroconidia 

are barrel-shaped. Appressoria absent. This species assimilates L-arabinose 

but not melibiose. Growth at 37 O C. Most common species, especially from invasive 

infections. RG-2 organism. 

Assimilation Tests: + Positive, - Negative, v Variable, w Weak, s Slow 

Glucose + Melibiose - L-Rhamnose + D-Glucitol v 

Galactose + Raffinose - D-Glucosamine + α-M-D-glucoside + 

L-Sorbose v Melezitose v N-A-D-glucosamine + D-Gluconate + 

Sucrose v Soluble Starch v Glycerol v DL-Lactate v 

Maltose + D-Xylose v Erythritol + myo-Inositol v 

Cellobiose + L-Arabinose + Ribitol v Nitrate - 

Trehalose + D-Arabinose + Galactitol - 2-K-D-gluconate + 

Lactose + D-Ribose + D-Mannitol v D-Glucuronate + 

Culture, hyphae and arthroconidia of Trichosporon asahii. 

20 µm 

Antifungal 

MIC µg/mL 


MIC µg/mL 

Range MIC90 Fluconazole 0.25-16 8.0 Amphotericin B 0.25-16 8.0 

Itraconazole 0.03-16 0.5 Flucytosine 2-128 16 

Posaconazole 0.06-16 1.0 Caspofungin >8 >8 

Voriconazole 0.03-16 0.25 Anidulafungin >8 >8 

MIC data for T. asahii. Antifungal susceptibility may vary between species and 

resistant strains have been reported. Therefore, antifungal susceptibility testing of 

individual strains is recommended. Paphitou et al. (2002), Espinel-Ingroff (2003), 

Rodriguez-Tudela et al. (2005), Metin et al. (2005) and WCH in-house data.

172 


Trichosporon asteroides (Rischin) Ota 

Colonies (SDA) are restricted, dry, cream-coloured, cerebriform, with radial furrows 

and irregular folds. The meristematic form is punctiform, brownish and consists of hyphae 

which swell and become multiseptate which may fall apart into smaller packets. 

Budding cells and lateral conidia are absent. Arthroconidia are elongate and hyphae 

are often present. Appressoria absent. This species assimilates L-arabinose but not 

myo-inositol. Growth at 37 O C is variable. Uncommon species usually associated with 

superficial infections. RG-2 organism. 



Galactose + Raffinose - D-Glucosamine v α-M-D-glucoside + 

L-Sorbose v Melezitose + N-A-D-glucosamine + D-Gluconate + 

Sucrose + Soluble Starch + Glycerol + DL-Lactate + 

Maltose + D-Xylose + Erythritol + myo-Inositol - 

Cellobiose + L-Arabinose + Ribitol v Nitrate - 

Trehalose + D-Arabinose + Galactitol - 2-K-D-gluconate + 


Trichosporon cutaneum (de Beurmann et al.) Ota 

Colonies (SDA) are cream-coloured, cerebriform, glabrous, with radial furrows and irregular 

folds. Budding cells abundant in primary cultures; hyphae developing in older 

cultures. Arthroconidia are cylindrical to ellipsoidal. Appressoria absent. This species 

assimilates melibiose; not tolerant to 0.1% cycloheximide. No growth at 37 O C. Uncommon 

species usually associated with superficial infections. RG-2 organism. 


Glucose + Melibiose + L-Rhamnose + D-Glucitol + 

Galactose + Raffinose + D-Glucosamine v α-M-D-glucoside + 



Maltose + D-Xylose + Erythritol + myo-Inositol + 

Cellobiose + L-Arabinose + Ribitol + Nitrate - 

Trehalose + D-Arabinose v Galactitol - 2-K-D-gluconate + 

Lactose + D-Ribose + D-Mannitol + D-Glucuronate + 

Trichosporon inkin (Oho ex Ota) do Carmo-Sousa & van Uden 

Colonies (SDA) are restricted, white, finely cerebriform with a granular covering, without 

marginal zone, often cracking the media. Budding cells and lateral conidia absent. 

Arthroconidia are long cylindrical. Appressoria present in slide cultures. Sarcinae 

present on media with high sugar-content. This species assimilates myo-inositol but 

not melibiose. Growth at 37 O C. Usually associated with white piedra on pubic hairs. 

RG-2 organism.


Trichosporon inkin (Oho ex Ota) do Carmo-Sousa & van Uden 


Glucose + Melibiose - L-Rhamnose - D-Glucitol - 

Galactose v Raffinose - D-Glucosamine v α-M-D-glucoside + 


Sucrose + Soluble Starch + Glycerol v DL-Lactate + 


Cellobiose + L-Arabinose v Ribitol - Nitrate - 

Trehalose + D-Arabinose v Galactitol - 2-K-D-gluconate + 


Trichosporon mucoides Gueho & M.Th. Smith 

Colonies (SDA) are moist and glabrous, white, cerebriform, heaped and folded. Budding 

cells present in primary cultures. Broadly clavate, terminal or lateral blastoconidia 

often present, becoming thick-walled with age. Arthroconidia are barrel-shaped. Appressoria 

absent. This species assimilates melibiose and is tolerant to 0.1% cycloheximide. 

Growth at 37 O C. Common species associated with superficial infections, white 

piedra and onychomycosis. RG-2 organism. 


Glucose + Melibiose + L-Rhamnose + D-Glucitol + 

Galactose + Raffinose + D-Glucosamine + α-M-D-glucoside + 

L-Sorbose + Melezitose + N-A-D-glucosamine + D-Gluconate + 



Cellobiose + L-Arabinose + Ribitol + Nitrate - 

Trehalose + D-Arabinose + Galactitol + 2-K-D-gluconate + 

Lactose + D-Ribose + D-Mannitol + D-Glucuronate + 

Trichosporon ovoides Behrend 

Colonies (SDA) are restricted, white, granular, folded at the centre, with a flat marginal 

zone. Budding cells and lateral conidia absent. Arthroconidia are cylindrical. Appressoria 

present in slide cultures. This species does not assimilate melibiose, but tolerates 

0.1% cycloheximide. Growth at 37 O C is variable. Uncommon species usually 

associated with superficial infections, like white piedra. RG-2 organism. 



Galactose + Raffinose v D-Glucosamine v α-M-D-glucoside + 

L-Sorbose v Melezitose v N-A-D-glucosamine + D-Gluconate + 

Sucrose + Soluble Starch + Glycerol v DL-Lactate + 


Cellobiose + L-Arabinose v Ribitol - Nitrate - 

Trehalose v D-Arabinose v Galactitol - 2-K-D-gluconate + 

Lactose + D-Ribose + D-Mannitol + D-Glucuronate +

174 


Trichothecium roseum (Persoon) Link ex Gray 

Colonies are moderately fast growing, flat, suede-like to powdery, initially white but 

becoming rosy, pink or orange with age. The conidiophores are indistinguishable 

from the vegetative hyphae until the first conidium is produced. They are erect, unbranched, 

often septate near the base, more or less rough-walled, bearing basipetal 

zig-zag (alternating) chains of conidia at the apex. Note: the conidiophore is progressively 

shortened with the formation of each conidium i.e. retrogressive conidial development. 

Conidia are two-celled ellipsoidal to pyriform, with an obliquely truncate basal 

scar, hyaline, smooth to delicately roughened and thick-walled. 

Trichothecium roseum has a world-wide distribution and is often isolated from decaying 

plant substrates, soil, seeds of corn, and food-stuffs (especially flour products). It 

is occasionally isolated as a saprophyte in the clinical laboratory. RG-1 organism. 

Trichothecium roseum should not be confused with Microsporum nanum. Colonies of 

the latter may be pinkish-buff in colour and also produce ovoid to pear-shaped, mostly 

two-celled macroconidia with thin, verrucose walls. However, M. nanum usually produces 

a red-brown reverse pigment and the two-celled macroconidia are sessile and 

formed singly, sometimes on stalks, on undifferentiated conidiophores which do not 

undergo further change or produce secondary conidia. Note: conidia are not produced 

in basipetal chains as in T. roseum. Finally, M. nanum will perforate hair in vitro. 

Key Features: hyphomycete, basipetal zig-zag chains of two-celled conidia showing 

retrogressive development where the conidiophore becomes progressively shorter. 


(1980), Domsch et al. (1980), Rippon (1988) and Samson et al. (1995). 

20 µm 

Conidiophores of T. roseum demonstrating retrogressive conidial development.


Ulocladium Preuss 

Colonies are rapid growing, brown to olivaceous-black or greyish and suede-like to 

floccose. Microscopically, numerous, usually solitary, multi-celled conidia (dictyoconidia) 

are formed through a pore (poroconidia) by a sympodially elongating geniculate 

conidiophore. Conidia are typically obovoid (narrowest at the base), dark brown and 

often rough-walled. Seven species have been described, all being saprophytes. RG- 

1 organism. 

Species of Ulocladium should not be confused with other poroconidial genera such as 

Stemphylium, Alternaria, Bipolaris, Exserohilum, Dreschlera and Curvularia. 


1976), Domsch et al. (1980), Rippon (1988), Samson et al. (1995) and de Hoog et al. 

(2000). 

Antifungal 

Conidia of Ulocladium. 

10 µm 

MIC µg/mL 

MIC µg/mL 

Antifungal 

Range Range 

Fluconazole 8->64 Amphotericin B 1->16 

Itraconazole 0.06->16 Flucytosine >128 

Voriconazole 0.25 

Very limited data. Antifungal susceptibility testing of individual strains is 

recommended. Pujol et al. (2000) and WCH in-house data.

176 


Veronaea botryosa Ciferri & Montemartini 

Colonies grow rapidly and are suede-like to downy, greyish-brown to blackish-brown. 

Conidiophores are erect, straight or flexuose, occasionally branched and are usually 

geniculate, due to the sympodial development of the conidia. They are smooth-walled, 

pale to medium olivaceous-brown, up to 250 µm long and 2-4 µm wide. Conidia are 

pale brown, two-celled, cylindrical with a truncated base, smooth-walled or slightly verrucose, 

5-12 x 3-4 µm. RG-1 organism. 

This genus is very similar to Rhinocladiella, however the conidia are typically twocelled. 

Occasional skin infections have been reported from humans. 

For descriptions of species, keys to taxa and additional information see Ellis (1971) 


Conidiophores and conidia of Veronaea botryosa. 

10 µm


Verticillium Nees ex Link 

Colonies are fast growing, suede-like to downy, white to pale yellow in colour, becoming 

pinkish brown, red, green or yellow with a colourless, yellow or reddish brown reverse. 

Conidiophores are usually well differentiated and erect, verticillately branched 

over most of their length, bearing whorls of slender awl-shaped divergent phialides. 

Conidia are hyaline or brightly coloured, mostly one-celled, and are usually borne in 

slimy heads (glioconidia). 

Members of this genus are often isolated from the environment. It has been reported 

as a rare agent of mycotic keratitis. RG-1 organism. 

Key Features: hyphomycete, verticillate branched conidiophores bearing whorls of 

awl-shaped, divergent phialides. 


(1980), McGinnis (1980), Rippon (1988), Samson et al. (1995), de Hoog et al. (2000). 

20 µm 

Conidiophores, phialides and conidia of Verticillium.

178 


Calcofluor White with 10% KOH. 

For the direct microscopic examination of skin scrapings, hairs, nails and other clinical 

specimens for fungal elements. This as a very sensitive method, however, a fluorescence 

microscope with the correct ultraviolet filters is required (Hageage and Harrington, 

1984; Hollander et al., 1984; Monheit et al., 1984). 

Solution A: Potassium hydroxide reagent. 

Potassium hydroxide 10 g 

Glycerine 10 mL 

Distilled water 80 mL 

Solution B: Calcofluor white reagent. 

Calcofluor white 0.5 g 

Evans blue 0.02 g 


Mix one drop of each solution on the centre of a clean microscope slide. 

Place the specimen in the solution and cover with a coverslip. 

Potassium Hydroxide (KOH) with Chlorazol Black. 

For the direct microscopic examination of skin scrapings, hairs, nails and other clinical 

specimens for fungal elements. Note: Parker Quink ink is no longer available. 

Potassium hydroxide 10 g 

Coral Azole E Black (0.1% - 100mg in 100ml) 10 mL 

Glycerol 10 mL 


Using sterile technique, remove a small portion of the specimen with an 

inoculation needle and mount in a drop of KOH on a clean microscope 

slide. Cover with a coverslip, squash the preparation with the butt of the 

inoculation needle and then blot off the excess fluid. 

Indian Ink Mounts. 

MICROSCOPY STAINS & TECHNIQUES 

For the direct microscopic examination of CSF for Cryptococcus species. Place a drop 

of Indian Ink on the specimen, mix well with a sterilised loop, and cover with a coverslip. 

Best brands to use are “Pelikan” or “Talons” Indian Ink.


Lactophenol Cotton Blue (LPCB). 

For the staining and microscopic identification of fungi. 

Cotton Blue (Aniline Blue) 0.05 g 

Phenol Crystals (C 6 H 5 O 4 ) 20 g 

Glycerol 40 mL 

Lactic acid (CH 3 CHOH COOH) 20 mL 


This stain is prepared over two days. 

1. On the first day, dissolve the Cotton Blue in the distilled water. Leave 

overnight to eliminate insoluble dye. 

2. On the second day, wearing gloves add the phenol crystals to the lactic 

acid in a glass beaker. Place on magnetic stirrer until the phenol is 

dissolved. 

3. Add the glycerol. 

4. Filter the Cotton Blue and distilled water solution into the phenol/glycerol/ 

lactic acid solution. Mix and store at room temperature. 

Direct Microscopic Mounts or Squash Preparations. 

Using sterile technique, remove a small portion of the colony with an inoculation needle 

and mount in a drop of Lactophenol Cotton Blue on a clean microscope slide. 

Cover with a coverslip, squash the preparation with the butt of the inoculation needle 

and then blot off the excess fluid. 

Cellotape Flag Preparations. 


An excellent technique for the rapid mounting of sporulating fungi because it keeps 

more of the reproductive structures intact. 

1. Using clear 2cm wide cellotape and a wooden applicator stick (orange 

stick) make a small cellotape flag (2 x 2 cm). 

2. Using sterile technique, gently press the sticky side of the flag onto the 

surface of the culture. 

3. Remove and apply a drop of 95% alcohol to the flag, this acts as a wetting 

agent and also dissolves the adhesive glue holding the flag to the 

applicator stick. 

4. Place the flag onto a small drop of Lactophenol cotton blue on a clean 

glass slide, remove the applicator stick and discard, add another drop 

of stain, cover with a coverslip, gently press and mop up any excess 

stain.

180 


Slide Culture Preparations. 


In order to accurately identify many fungi it is essential to observe the precise arrangement 

of the conidiophores and the way in which spores are produced (conidial ontogeny). 

Riddel’s simple method of slide culturing (Mycologia 42:265, 1950) permits fungi 

to be studied virtually in situ with as little disturbance as possible. A simple modification 

of this method using a single agar plate is described below. 

One plate of nutrient agar; potato dextrose is recommended, however, 

some fastidious fungi may require harsher media to induce sporulation like 

Cornmeal agar or Czapek Dox agar. 

1. Using a sterile blade cut out an agar block (7 x 7 mm) small enough to 

fit under a coverslip. 

2. Flip the block up onto the surface of the agar plate. 

3. Inoculate the four sides of the agar block with spores or mycelial 

fragments of the fungus to be grown. 

4. Place a flamed coverslip centrally upon the agar block. 

5. Incubate the plate at 26 O C until growth and sporulation have 

occurred. 

6. Remove the cover slip from the agar block. 

7. Apply a drop of 95% alcohol as a wetting agent. 

8. Gently lower the coverslip onto a small drop of Lactophenol cotton blue 

on a clean glass slide. 

9. The slide can be left overnight to dry and later sealed with fingernail 

polish. 

10. When sealing with nail polish use a coat of clear polish followed by one 

coat of red-coloured polish. 

Simple agar block method, inoculated on four sides with cover slip 

on top. Make at least 2 slides per culture.


Bird Seed Agar (Staib, 1987). 

for selective isolation of Cryptococcus neoformans and C. gattii. 

Guizotia abyssinica (niger seed) 50 g Glucose 1 g 

KH PO (potassium dihydrogen 1 g Creatinine 1 g 

2 4 

orthophosphate) 

Bacto Agar (BD 214010) 15 g Distilled water 1000 mL 

Penicillin G (20 units/mL) 1 mL Gentamicin (40 mg/mL) 1 mL 

1. Grind seeds of Guizotia abyssinica as finely as possible with an electric mixer 

and add to 1000 mL distilled water in a stainless steel jug. 

2. Boil for 30 minutes, pass through filter paper and adjust volume to 1000 mL. 

3. Add remaining ingredients except Bacto Agar to filtrate and dissolve. 

If required: Cool to room temperature and adjust pH to 5.5. 

Dispense into 500 mL bottles. 

4. Add 7.5 g Bacto Agar to each 500 mL reagent bottle. 

5. Autoclave 110OC for 20 minutes. 

6. Cool to 48OC and add 0.5 mL Penicillin G and 0.5 mL Gentamicin to each 500 

mL of Bird Seed Agar. 

7. Mix gently and pour into 90 mm plastic petri dishes. 

Bromcresol Purple Milk Solids Glucose Agar (BCP-MS-G). 

for the differentiation of Trichophyton species (Kane et al. 1977). 

Solution A: 


Skim milk powder (Carnation Brand) 80 g 

Bromcresol (or bromocresol) purple 

2 mL 

(1.6% solution in alcohol) 

Dissolve in 2 litre flask and autoclave 10 psi/15 minutes. 

Solution B: 

Glucose 40 g Distilled water 200 mL 

Dissolve and autoclave at 10 psi/8minutes. 

Solution C: 

SPECIALISED CULTURE MEDIA 

Bacto Agar (BD 214010) 30 g Distilled water 800 mL 

Soak for 15 minutes in 3 litre flask; autoclave at 15 psi/15 minutes 

To make media; add solution A and B to solution C. Adjust final pH to 6.6. 

Aseptically dispense for slopes (7 mL amounts into 30 mL disposable bottles). 

Caution: Do not substitute casein for skim milk. Check pH is 6.6.

182 


CDBT (Creatinine dextrose bromothymol blue thymine agar). 

for differentiation of Cryptococcus neoformans var. neoformans and 

Cryptococcus neoformans var. grubii (Irokanulo et al. 1994). 

Solution A: 

Creatinine 1 g Dextrose 0.5 g 

KH 2 PO 4 1 g MgSO 4 7H 2 O 0.5 g 

Thymine 0.1 g Distilled water 980 mL 

1. Dissolve ingredients in small beaker and adjust pH to 5.6 

2. Store in refrigerator. 

Solution B (Aqueous Bromothymol Blue): 

Bromothymol blue 0.4 g 0.01N NaOH 64 mL 


1. Dissolve the Bromothymol Blue in the NaOH 

2. Add to the water. 

To prepare medium (1 litre for plates): 

Solution A 980 mL Solution B 20 mL 

Bacto Agar (BD 214010) 20 g 

Autoclave to 121 O C for 15 minutes, cool to 48 O C and pour plates. 

CGB (L-Canavanine glycine bromothymol blue agar). 

for differentiation of Cryptococcus neoformans and Cryptococcus gattii 

(Kwon-Chung et al. 1982). 

Solution A: 

Glycine Univar 10 g KH 2 PO 4 1 g 

MgSO 4 1 g Thiamine HCl 1 mg 

L-canavanine sulphate 30 mg Distilled water 100 mL 

1. Dissolve ingredients in small beaker and adjust pH to 5.6 

2. Filter sterilise solution using 0.22 µm filter. 

3. Store in refrigerator. 

Solution B (Aqueous Bromothymol Blue): 

Bromothymol blue 0.4 g 0.01N NaOH 64 mL 


1. Dissolve the Bromothymol Blue in the NaOH 

2. Add to the water. 

To prepare medium (1 litre for plates): 


Distilled water 980 mL Solution B 20 mL 


1. Autoclave to 121 O C for 15 minutes, cool to 48 O C. 

2. For plates add 100 mL of the filtered solution A and mix. Dispense in 

plates.



Cornmeal Agar. 

for routine cultivation and identification of fungi. 

Cornmeal agar (Oxoid CM 0103) 8.5 g 


1. Mix dry ingredients into 100 mL H 2 O, boil remaining water. 

2. Add boiling water to mixture and bring to boil. 

3. Autoclave for 10 minutes at 120 O C, then slope on racks. 

Cornmeal Glucose Sucrose Yeast Extract Agar. 

for zygomycete sporulation 

Cornmeal agar (Oxoid CM 0103) 17 g 

Dextrose (Glucose) 2 g 

Sucrose 3 g 

Yeast extract 1 g 


1. Mix dry ingredients into 100 mL H 2 O, boil remaining water. 

2. Add boiling water to mixture and bring to boil. 

3. Dispense for slopes. 

4. Autoclave for 10 minutes at 120 O C, remove and slope. 

Czapek Dox Agar. 

for routine cultivation of fungi, especially Aspergillus, Penicillium, 

and non-sporulating moulds. 

Czapek Dox Agar (Oxoid CM97) 45.4 g 


1. Soak the ingredients in small amount of water. 

2. Bring remaining water to boil, add to soaking ingredients and bring to 

the boil again, stirring continuously. 

3. Dispense for slopes as required. 

4. Autoclave at 121 O C for 10 minutes, remove and slope or pour for plates 

as required. 

Dixon’s Agar (modified). 

for primary isolation and cultivation of Malassezia species. 

Malt extract (Oxoid L39) 9 g Bacto Tryptone 1.5 g 

Ox-bile Desiccated (Oxoid L50) 5 g Tween 40 2.5 mL 

Oleic acid 0.5 g Glycerol 0.5 mL 

Bacto Agar 3 g Distilled water 250 mL 

1. Soak ingredients in a little of the water. 

2. Bring remaining water to boil, add to the soaking ingredients and bring 

to the boil again constantly stirring. 

3. Dispense for slopes (7 mL amounts into 30 mL disposable bottles. 

4. Autoclave at 121 O C for 10 minutes and then slope.

184 



Hair Perforation Test. 

for the differentiation of Trichophyton species. 

Blonde pre-pubital hair cut into short pieces (1 cm) 10-20 hairs 


1. Autoclave hair at 121 O C for 10 minutes and store in sterile container. 

2. Place 10-20 short pieces of hair in 5 mL water in vial. 

3. Inoculate with small fragments of the test fungus. 

4. Incubate at room temperature. 

5. Individual hairs are removed at intervals up to 4 weeks and examined 

microscopically in lactophenol cotton blue. Isolates of T. mentagrophytes 

produce marked localised areas of pitting and marked erosion whereas 

those of T. rubrum do not. 

Lactritmel Agar. 

for the production of pigment by Trichophyton species. 

Skimmed milk powder 

7 g 

(use only Dutch Jug skimmed milk powder) 

Honey 10 g 

Cornmeal agar (Oxoid CM 0103) 17 g 

Chloramphenicol 1 x 250 capsule 


1. Weigh skimmed milk into stainless steel jug. Slowly add some water, 

mixing milk into smooth paste. Continue adding small quantities of 

water until powder is dissolved (about 150 mL). 

2. Weigh other ingredients into skimmed milk and allow to soak. 

3. Boil remaining water, and with it wash out honey from beaker. 

4. Add to other ingredients and boil. 

5. Dispense for slopes (7 mL). 

6. Autoclave for 10 minutes at 115OC. 7. On removal from autoclave allow to stand 5 minutes then shake and 

slope on racks. 

Note: Do not filter or adjust pH in any way 

Littman Oxgall Agar. 


Littman Oxgall Agar (US Biological L3025) 27.5 g 


1. Soak agar in 100 mL of water in stainless steel jug. Boil remaining 

400mL in a separate jug. 

2. When water has boiled add to soaking agar and reboil, stirring 

constantly. 


4. Autoclave for 10 minutes at 121 O C, remove and slope.



Malt Extract Agar. 


Oxoid Malt Extract (L39) 20 g 



1. Dissolve malt extract in a plastic beaker and pH the solution to pH 6.5 

with NaOH. 

2. Soak agar in small quantity of solution. Bring remaining solution to 

the boil, stirring constantly. 

3. Add to soaking agar. Bring to boil, stirring constantly. 


5. Autoclave at 121 O C for 10 minutes, remove and slope or pour for 

plates as required. 

1% Peptone Agar. 


Tryptone Peptone (BD 211705) 5 g 



1. Soak agar and peptone in about 50 mL of water. 

2. Boil remaining water, add this to soaking ingredients and bring to boil 

again. 


4. Autoclave for 10 minutes at 121 O C, then slope on racks. 

Potato Dextrose Agar. 


Potato Dextrose Agar (Oxoid CM139) 39 g 


1. Soak potato dextrose agar in small amount of the water in a stainless 

steel jug. 

2. Boil remaining water, add to soaking ingredients, bring to the boil, 

stirring constantly. 


4. Autoclave at 121 O C for 15 minutes. Remove and slope or pour for 

plates as required. 

Rice Grain Slopes. 

to induce sporulation and for differentiation of M. audouinii and M. canis. 

Polished rice grains Distilled water 

1. Place ~ 1/2 teaspoon rice grains into wide neck 20 mL glass vials. 

2. Add 8 mL distilled water to each vial. 

3. Lid, then slope on racks ensuring rice grains are evenly distributed. 

4. Autoclave racks at 121 O C for 15 minutes.

186 



Sabouraud Dextrose Agar with Cycloheximide, Chloramphenicol, 

Gentamicin and Yeast Extract. 

for the primary isolation and cultivation of dermatophytes. 

Sabouraud Dextrose Agar (Oxoid CM41) 65 g 

Cycloheximide (Actidione) 0.5 g 


Gentamicin (40mg/mL) 0.56 mL 

Yeast extract 5 g 


1. Soak all ingredients, except Gentamicin, in 100 mL water. 

2. Boil remaining water, add to soaking ingredients, and bring to boil to 

dissolve, stirring well to prevent from charring. 

3. Add the Gentamicin. Mix well. 


5. Autoclave at 121 O C for 10 minutes. Remove and slope, or pour for plates 

as required. 

Sabouraud Dextrose Agar with Chloramphenicol and Gentamicin. 

for primary isolation and routine culture of yeasts and moulds. 

Sabouraud Dextrose Agar (Oxoid CM41) 65 g 


Gentamicin (40mg/mL) 0.56 mL 


See above method for Sabouraud Dextrose Agar with Cycloheximide, 

Chloramphenicol, Gentamicin and Yeast Extract. 

Sabouraud Dextrose Agar with 5% Salt. 


Sabouraud Dextrose Agar (Oxoid CM41) 32.5 g 

Sodium Chloride NaCl (Univar 465) 25 g 


1. Soak ingredients in approximately 100 mL water. 

2. Bring remaining water to boil, add to soaking ingredients. 


4. Autoclave at 118 O C for 10 minutes, then slope on racks.


Tap Water Agar. 

for the stimulation of sporulation in Apophysomyces and Saksenaea isolates. 



1. Add agar to water in stainless steel jug, allow to soak. 


3. Autoclave at 118 O C for 10 minutes, remove and slope. 

Urease Agar Slopes with 0.5% Glucose. 

for the differentiation of Urease producing organisms. 

Urease glucose broth base: 

Urea, broth base (Oxoid CM71) 0.9 g 

Glucose 5 g 


1. Add the Urea broth base and glucose to the distilled water in a 500mL 

beaker. 

2. Dispense in 5 X 90 mL amounts. 

3. Autoclave at 115 O C for 20 mins. 

4. When cool, label and store in the fridge. 

Method to make slopes: 


40% Urea Solution (Oxoid SR 20) 10 mL 



1. Add 3.0 grams of agar to 100 mL of distilled water in a 250 mL pyrex 

bottle. 

2. Autoclave at 121 O C for 15 minutes and place in 50 O C water bath. 

3. When cool add 90 mL of the Urease broth with glucose and the 10 mL 

of 40% urea solution to agar and dispense in 3 mL aliquots and slope 

on racks. 

Vitamin Free Agar (Trichophyton Agar No.1). 


Trichophyton Agar No. 1 (BD 287710) 11.8 g 


1. Add agar to water in stainless steel jug, allow to soak. 

2. Bring to boil to dissolve, stirring constantly. 

3. Once boiled remove immediately to avoid discolouration. 


5. Autoclave at 118 O C for 10 minutes, remove and slope.

188 


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