IDENTIFICATION OF PATHOGENIC FUNGI

IDENTIFICATION OF PATHOGENIC FUNGIColin K. Campbell PhDHealth Protection Agency Mycology Reference LaboratoryBristol, UK (retired)

Elizabeth M. Johnson PhDHealth Protection Agency Mycology Reference LaboratoryBristol, UK

David W. Warnock PhD, FAAM, FRCPathNational Center for Emerging and Zoonotic Infectious DiseasesCenters for Disease Control and PreventionAtlanta, Georgia, USA

SECOND EDITION

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DisclaimerThis text was co-authored by David W. Warnock in his private capacity. No official support or endorsement by the U.S. Centers for Disease Control and Prevention or by the U.S. Department of Health and Human Services is intended or should be inferred.

This edition first published 2013. © 1996, 2013 Health Protection Agency

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Library of Congress Cataloging-in-Publication DataCampbell, Colin K. Identification of pathogenic fungi / Colin Campbell, Elizabeth Johnson, David W. Warnock. – 2nd ed. p. ; cm. Rev. ed. of: Identification of pathogenic fungi / Colin K. Campbell ... [et al.]. London : Public Health Laboratory Service, c1996. Includes bibliographical references and index. ISBN 978-1-4443-3070-0 (hardback) I. Johnson, Elizabeth M., Ph. D. II. Warnock, D. W. III. Identification of pathogenic fungi. IV. Title. [DNLM: 1. Fungi–isolation & purification. 2. Fungi–pathogenicity. 3. Mycoses–diagnosis. QW 180]

579.5–dc23 2012044506

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v

Preface, ixAcknowledgements, xi

  1  Introduction, 1

  2  Identification of Moulds, 11Media for Mould Identification, 14Mounting Fluids, 16

  3  Moulds with Arthrospores, 18Neoscytalidium dimidiatum, 20Coccidioides species, 24Onychocola canadensis, 28

  4  Moulds with Aleuriospores: I. The Dermatophytes, 31Microsporum gypseum, 38Microsporum canis, 40Microsporum equinum, 42Epidermophyton floccosum, 44Trichophyton terrestre, 46Trichophyton rubrum, 48Trichophyton interdigitale, 52Trichophyton mentagrophytes, 54Trichophyton erinacei, 56Trichophyton equinum, 58Trichophyton soudanense, 60Microsporum persicolor, 62Trichophyton tonsurans, 64Microsporum audouinii, 66Trichophyton violaceum, 68Trichophyton verrucosum, 70Trichophyton schoenleinii, 72Trichophyton concentricum, 74Other Microsporum and Trichophyton species, 76

  5  Moulds with Aleuriospores: II. Others, 80Geomyces pannorum, 82Chrysosporium keratinophilum, 84Myceliophthora thermophila, 86

CONTENTS

vi

CONTENTS

Histoplasma capsulatum, 88Blastomyces dermatitidis, 92Paracoccidiodes brasiliensis, 96

  6  Moulds with Holoblastic Conidia, 99Aureobasidium pullulans, 102Sporothrix schenckii, 104Cladophialophora bantiana, 106Cladosporium sphaerospermum, 108Fonsecaea pedrosoi, 110Rhinocladiella atrovirens, 112Rhinocladiella mackenziei, 114Ochroconis gallopava, 116Alternaria alternata, 118Ulocladium chartarum, 120Curvularia lunata, 122Bipolaris hawaiiensis, 124Exserohilum rostratum, 126

  7  Moulds with Enteroblastic Conidia Adhering in Chains, 129Aspergillus flavus species complex, 134Aspergillus fumigatus species complex, 136Aspergillus glaucus, 138Aspergillus nidulans species complex, 140Aspergillus versicolor species complex, 142Aspergillus ustus species complex, 144Aspergillus niger species complex, 146Aspergillus terreus species complex, 148Aspergillus candidus species complex, 150Penicillium marneffei, 152Scopulariopsis brevicaulis, 154Purpureocillium lilacinum, 156Paecilomyces variotii, 158

  8  Moulds with Enteroblastic Conidia Adhering in Wet Masses, 161Fusarium lichenicola, 166Fusarium dimerum species complex, 168Fusarium semitectum, 170Fusarium proliferatum, 172

vii

CONTENTS

Fusarium oxysporum species complex, 174Fusarium solani species complex, 176Acremonium strictum, 178Acremonium kiliense, 180Lecythophora mutabilis, 182Scedosporium prolificans, 184Scedosporium apiospermum, 186Phaeoacremonium parasiticum, 188Pleurostomophora richardsiae, 190Phialophora verrucosa, 192Hortaea werneckii, 194Exophiala spinifera, 196Exophiala dermatitidis, 198Exophiala jeanselmei, 200

  9  Mucoraceous Moulds and Their Relatives, 203Cunninghamella bertholletiae, 208Lichtheimia corymbifera, 210Rhizomucor pusillus, 212Mucor circinelloides, 214Rhizopus microsporus, 216Rhizopus arrhizus, 218Mucor hiemalis, 220Basidiobolus ranarum, 222Conidiobolus coronatus, 224Pythium insidiosum, 226Apophysomyces elegans, 228Saksenaea vasiformis, 230Mortierella wolfii, 232

10  Miscellaneous Moulds, 235Aphanoascus fulvescens, 238Monascus ruber, 240Chaetomium species, 242Phoma herbarum, 244Myxotrichum deflexum, 246Schizophyllum commune, 248Leptosphaeria senegalensis, 250Neotestudina rosatii, 252

viii

CONTENTS

Piedraia hortae, 254Lasiodiplodia theobromae, 256Pyrenochaeta romeroi, 258Madurella mycetomatis, 260

11  Identification of Yeasts, 263Media for Yeast Identification, 272Candida albicans, 274Candida tropicalis, 276Candida krusei, 278Candida lipolytica, 280Candida kefyr, 281Candida lusitaniae, 282Candida parapsilosis, 284Candida pelliculosa, 286Candida guilliermondii, 287Candida glabrata, 288Cryptococcus neoformans and Cryptococcus gattii, 290Rhodotorula glutinis, 292Saccharomyces cerevisiae, 294Geotrichum candidum, 296Saprochaete capitata, 298Trichosporon species, 300Malassezia furfur species complex, 302Malassezia pachydermatis, 304

12  Identification of Fungi in Sections, Smears and Body Fluids, 305

Appendix 1:  Common Mycological Terms, 321Appendix 2:  Further Reading, 325

Species Index, 327Subject Index, 333

ix

In the seventeen years that have elapsed since the first edition of this manual was pub-lished, much progress has been made in the diagnosis, treatment and prevention of fungal diseases. Nonetheless, these infections continue to be a leading cause of serious illness and death in many different patient populations. New methods of diagnosis have been intro-duced and new antifungal agents have been licensed for use, but these developments have been offset by the emergence of resistance to several classes of drugs and an increase in infections caused by fungi with innate resistance to one or more classes.

Many of the moulds that are now recognized as being capable of producing serious disease in immunocompromised or debilitated individuals are environmental organisms whose natural habitat is in the soil or on plants, wood, compost heaps, or rotting food. Most are familiar to mycologists, plant pathologists and food microbiologists, but they present problems for the clinical microbiologist who often has had no formal training in the identification of fungi. This process can be challenging and sometimes frustrating because of the importance placed on the morphological characteristics of the organisms, and the need to become familiar with a wide range of different structures and terms.

As before, this manual has been designed for use by medical, scientific and technical staff in hospital laboratories in the UK and abroad, but we hope it will also be of interest to other groups of scientists. The organisms described have been grouped in chapters accord-ing to spore-bearing structures produced in culture, rather than simply being ordered on an alphabetical basis. Each chapter has been arranged so that the descriptions for similar organisms may be found on adjacent pages. In addition, we have attempted to provide differential diagnoses on the basis of both colonial appearance and microscopic character-istics for the organisms described. Although molecular methods are assuming an ever greater importance, routine identification of moulds still rests, for the most part, on mor-phological examination. To assist with this, we have added colour illustrations of cultures and microscopic structures to the line drawings that are found throughout the manual. Lack of space has precluded the inclusion of every rare organism that might be isolated from a clinical specimen. In some cases a single representative member of a genus is described, and isolates that appear similar to the description provided may need to be referred to a specialist for confirmation of the identification.

For this edition, we have added a new chapter on the identification of fungi in his-topathogical sections and smears. As before, we have included two appendices: the first giving definitions of many mycological terms in common use and the second listing some useful monographs and more comprehensive texts that the reader may wish to consult.

Colin K. CampbellElizabeth M. Johnson

David W. Warnock

PREFACE

xi

We are very grateful to our colleagues, Dr Andrew Borman, Mrs Adrien Szekely and Dr Christopher Linton from the Mycology Reference Laboratory, Bristol, for their input during the development of this new edition, and for their many helpful suggestions for its improve-ment. We are also grateful to our late colleague Dr Christine Philpot for her involvement in the first edition. We also wish to thank Kate Newell of Wiley-Blackwell and project manager Kathy Syplywczak for their invaluable help in the design and production of this monograph.

Colin K. CampbellElizabeth M. Johnson

David W. Warnock

ACKNOWLEDGEMENTS

1

The kingdom Fungi consists of a distinct group of eukaryotic organisms that absorb their nourishment from living or dead organisms or organic matter. Fungi are found throughout nature, performing an essential service in returning to the soil nutrients removed by plants. There is, however, a large group of species that are parasitic on plants and a smaller group that are parasitic on animals, as well as on man. Fungi show considerable variation in size and form, but can be divided into three main groups: multicellular filamentous fungi (moulds); unicellular fungi (yeasts); and dimorphic fungi which are capable of changing their growth to either a multicellular or unicellular form, depending on the growth conditions.

In most multicellular fungi, the vegetative stage consists of a system of tubular, branch-ing filaments, or mycelium. Each individual filament, or hypha, has a rigid cell wall and increases in length as a result of apical growth. In the more primitive fungi, the hyphae remain aseptate (without cross-walls). In the more advanced groups however, the hyphae are divided into compartments or cells by the development of more or less frequent cross-walls, termed septa. Such hyphae are termed septate.

Yeasts are unicellular fungi consisting of separate, round, oval or elongated cells or blastospores that propagate by an asexual process called budding in which the cell develops a protuberance from its surface. The bud enlarges and may become detached from the parent cell, or it may remain attached and itself produce another bud. In this way a chain of cells may be produced. Under certain conditions, continued elongation of the parent cell before it buds results in a chain of elongated cells, termed a pseudohypha, which resembles the hypha of moulds. Unlike a true hypha, however, the connection between adjacent pseudohyphal cells shows a marked constriction. Some yeasts can also produce true hyphae, with cross-walls. A small number of yeasts reproduce by fission. Yeasts are neither a natural nor a formal taxonomic group, but are a growth form shown by a wide range of unrelated fungi.

Some medically important fungi change their growth form during the process of tissue invasion. These dimorphic pathogens usually change from a multicellular hyphal form in the natural environment to a budding, single-celled yeast form in tissue.

Fungi reproduce by means of microscopic propagules, termed spores, that consist of a single cell or several cells contained within a rigid wall. Spores may be produced by an asexual process (involving mitosis only) or by sexual reproduction (involving meiosis). Some species of fungi are homothallic and able to form sexual structures within individual colonies. Most, however are heterothallic and do not form their sexual structures unless

1  INTRODUCTION

Identification of Pathogenic Fungi, Second Edition. Colin K. Campbell, Elizabeth M. Johnson, and David W. Warnock.© 2013 Health Protection Agency. Published 2013 by Blackwell Publishing Ltd.