Larone's Medically Important Fungi

The definitive guide for identifying fungi from clinical specimens

Medically Important Fungi will expand your knowledge and support your work by:

• Providing detailed descriptions of the major mycoses as viewed in patients' specimens by direct microscopic examination of stained slides
• Offering a logical step-by-step process for identification of cultured organisms, utilizing detailed descriptions, images, pointers on organisms' similarities and distinctions, and selected references for further information
• Covering nearly 150 of the fungi most commonly encountered in the clinical mycology laboratory
• Presenting details on each organism's pathogenicity, growth characteristics, relevant biochemical reactions, and microscopic morphology, illustrated with photomicrographs, Dr. Larone's unique and elegant drawings, and color photos of colony morphology and various test results
• Explaining the current changes in fungal taxonomy and nomenclature that are due to information acquired through molecular taxonomic studies of evolutionary fungal relationships
• Providing basic information on molecular diagnostic methods, e.g., PCR amplification, nucleic acid sequencing, MALDI-TOF mass spectrometry, and other commercial platforms
• Including an extensive section of easy-to-follow lab protocols, a comprehensive list of media and stain procedures, guidance on collection and preparation of patient specimens, and an illustrated glossary

With Larone's Medically Important Fungi: A Guide to Identification, both novices and experienced professionals in clinical microbiology laboratories can continue to confidently identify commonly encountered fungi.

If you are looking for online access to the latest clinical microbiology content, please visit www.wiley.com/learn/clinmicronow. 

Davise H. Larone is well known as the originator of the book that many readers have come to rely upon for assistance in the accurate identification of fungi from patient specimens, a key step in treating mycotic infections. Dr. Larone has now been joined by Thomas J. Walsh and Randall T. Hayden to update this gold standard reference while retaining the format that has made this guide so popular for more than 40 years.

List of Tables xv

Preface to the Fifth Edition xvii

Preface to the First Edition xix

Acknowledgments xxi

How to Use the Guide 1

Use of Reference Laboratories 3

Safety Precautions 7

Part I Direct Microscopic Examination of Clinical Specimens

Introduction 11

Histological Terminology 13

Tissue Reactions to Fungal Infection 17

Stains 21

Table 1 Stains for direct microscopic observation of fungi and/or filamentous bacteria in tissue 22

Guide to Interpretation of Direct Microscopic Examination 23

Detailed Descriptions 31

Actinomycosis 33

Mycetoma, Actinomycotic or Eumycotic 34

Nocardiosis 36

Zygomycosis 37

Aspergillosis 38

Miscellaneous Hyalohyphomycoses 40

Dermatophytosis 42

Tinea versicolor 43

Tinea nigra 44

Phaeohyphomycosis 45

Chromoblastomycosis 46

Sporotrichosis 47

Histoplasmosis capsulati 48

Penicilliosis marneffei 50

Blastomycosis 52

Paracoccidioidomycosis 53

Candidiasis (Candidosis) 54

Trichosporonosis 56

Cryptococcosis 57

Pneumocystosis 59

Protothecosis 60

Coccidioidomycosis 61

Rhinosporidiosis 62

Adiaspiromycosis 64

Special References 65

Part II Identification of Fungi in Culture

Guide to Identification of Fungi in Culture 69

Detailed Descriptions 101

Filamentous Bacteria 103

Introduction 105

Table 2 Differentiation of filamentous aerobic actinomycetes encountered in clinical specimens 107

Nocardia spp. 108

Table 3 Phenotypic characteristics of most common clinically encountered Nocardia spp. 110

Streptomyces spp. 111

Actinomadura spp. 112

Nocardiopsis dassonvillei 113

Yeasts and Yeastlike Organisms 115

Introduction 117

Candida albican 119

Table 4 Characteristics of the genera of clinically encountered yeasts and yeastlike organisms 120

Candida dubliniensis 121

Table 5 Characteristics of Candida spp. most commonly encountered in the clinical laboratory 122

Table 6 Characteristics that assist in differentiating Candida dubliniensis from Candida albicans 124

Candida tropicalis 125

Candida parapsilosis complex 126

Candida lusitaniae 127

Candida krusei 128

Table 7 Differentiating characteristics of Blastoschizomyces capitatus versus Candida krusei 129

Table 8 Differentiating characteristics of C. krusei, C. inconspicua, and C. norvegensis 129

Candida kefyr (formerly Candida pseudotropicalis) 130

Candida rugosa 131

Candida guilliermondii complex 132

Table 9 Differentiating characteristics of Candida guilliermondii versus Candida famata 133

Candida lipolytica 134

Candida zeylanoides 135

Candida glabrata 136

Cryptococcus neoformans 137

Cryptococcus gattii 138

Table 10 Characteristics of Cryptococcus spp. 139

Table 11 Characteristics of yeasts and yeastlike organisms other than Candida spp. and Cryptococcus spp. 140

Rhodotorula spp. 141

Sporobolomyces salmonicolor 142

Saccharomyces cerevisiae 143

Wickerhamomyces anomalus (formerly Pichia anomala and Hansenula anomala) (sexual state); Candida pelliculosa (asexual state) 145

Malassezia spp. 146

Malassezia pachydermatis 148

Ustilago sp. 149

Prototheca spp. 150

Trichosporon spp. 151

Table 12 Key characteristics of the most common clinically encountered Trichosporon spp. 152

Blastoschizomyces capitatus 153

Geotrichum candidum 154

Thermally Dimorphic Fungi 155

Introduction 157

Histoplasma capsulatum 158

Blastomyces dermatitidis 160

Paracoccidioides brasiliensis 162

Penicillium marneffei 164

Sporothrix schenckii complex 166

Table 13 Characteristics for differentiating species of the Sporothrix schenckii complex 168

Thermally Monomorphic Moulds 169

Zygomycetes 171

Introduction 173

Table 14 Differential characteristics of similar organisms in the class Zygomycetes 175

Table 15 Differential characteristics of the clinically encountered Rhizopus spp. 175

Rhizopus spp. 176

Mucor spp. 177

Rhizomucor spp. 178

Lichtheimia corymbifera complex (formerly Absidia corymbifera) 179

Apophysomyces elegans 181

Saksenaea vasiformis 183

Cokeromyces recurvatus 184

Cunninghamella bertholletiae 185

Syncephalastrum racemosum 186

Basidiobolus sp. 187

Conidiobolus coronatus 188

Dematiaceous Fungi 189

Introduction 191

Fonsecaea pedrosoi 193

Fonsecaea compacta 195

Rhinocladiella spp. 196

Phialophora verrucosa 197

Table 16 Characteristics of Phialophora, Pleurostomophora, Phaeoacremonium, Acremonium, Phialemonium, and Lecythophora 198

Pleurostomophora richardsiae (formerly Phialophora richardsiae) 199

Phaeoacremonium parasiticum (formerly Phialophora parasitica) 200

Phialemonium spp. 201

Cladosporium spp. 203

Table 17 Characteristics of Cladosporium and Cladophialophora spp. 204

Cladophialophora carrionii 205

Cladophialophora bantiana 206

Cladophialophora boppii (formerly Taeniolella boppii) 207

Pseudallescheria boydii (sexual state) / Scedosporium apiospermum (asexual state) complex 208

Table 18 Differentiating phenotypic characteristics of the clinically encountered members of the Pseudallescheria boydii complex and Scedosporium prolificans 210

Scedosporium prolificans (formerly Scedosporium inflatum) 211

Ochroconis gallopava (formerly Dactylaria constricta var.gallopava) 212

Table 19 Differentiation of the clinically encountered Ochroconis species 213

Table 20 Characteristics of some of the “black yeasts” 213

Exophiala jeanselmei complex 214

Exophiala dermatitidis (Wangiella dermatitidis) 215

Hortaea werneckii (Phaeoannellomyces werneckii) 216

Madurella mycetomatis 217

Madurella grisea 218

Piedraia hortae 219

Aureobasidium pullulans 220

Table 21 Differential characteristics of Aureobasidium pullulans versus Hormonema dematioides 222

Hormonema dematioides 223

Neoscytalidium dimidiatum (formerly Scytalidium dimidiatum) 224

Botrytis sp. 226

Stachybotrys chartarum (S. alternans, S. atra) 227

Graphium eumorphum 228

Curvularia spp. 229

Bipolaris spp. 230

Table 22 Characteristics of Bipolaris, Drechslera, and Exserohilum spp. 231

Exserohilum spp. 232

Helminthosporium sp 233

Alternaria sp 234

Ulocladium sp. 235

Stemphylium sp. 236

Pithomyces sp. 237

Epicoccum sp. 238

Nigrospora sp. 239

Chaetomium sp. 240

Phoma spp. 241

Dermatophytes 243

Introduction 245

Microsporum audouinii 247

Microsporum canis var. canis 248

Microsporum canis var. distortum 249

Microsporum cookei 250

Microsporum gypseum complex 251

Microsporum gallinae 252

Microsporum nanum 253

Microsporum vanbreuseghemii 254

Microsporum ferrugineum 255

Trichophyton mentagrophytes 256

Table 23 Differentiation of similar conidia-producing Trichophyton spp 257

Trichophyton rubrum 258

Trichophyton tonsurans 259

Trichophyton terrestre 260

Trichophyton megninii 261

Trichophyton soudanense 262

Table 24 Growth patterns of Trichophyton species on nutritional test media 263

Trichophyton schoenleinii 264

Trichophyton verrucosum 265

Trichophyton violaceum 266

Trichophyton ajelloi 267

Epidermophyton floccosum 268

Hyaline Hyphomycetes 269

Introduction 271

Coccidioides spp. 272

Table 25 Differential characteristics of fungi in which arthroconidia predominate 274

Malbranchea spp. 275

Geomyces pannorum 276

Arthrographis kalrae 277

Hormographiella aspergillata 278

Emmonsia spp. 279

The Genus Aspergillus 281

Aspergillus fumigatus 283

Aspergillus niger 284

Aspergillus flavus 285

Table 26 Identification of the most common species of Aspergillus 286

Aspergillus versicolor 288

Aspergillus calidoustus 289

Aspergillus nidulans (asexual state); Emericella nidulans (sexual state) 290

Aspergillus glaucus (asexual state); Eurotium herbariorum (sexual state) 291

Aspergillus terreus 292

Aspergillus clavatus 293

Penicillium spp. 294

Paecilomyces spp. 295

Scopulariopsis spp. 297

Table 27 Differential characteristics of Paecilomyces variotii versus P. lilacinus 299

Table 28 Differential characteristics of Scopulariopsis brevicaulis versus S. brumptii 299

Gliocladium sp. 300

Trichoderma sp. 301

Beauveria bassiana 302

Verticillium sp. 303

Acremonium (formerly Cephalosporium) spp. 304

Fusarium spp. 305

Lecythophora spp. 307

Trichothecium roseum 308

Chrysosporium spp. 309

Table 29 Differential characteristics of Chrysosporium versus Sporotrichum 311

Sporotrichum pruinosum 312

Sepedonium sp. 313

Chrysonilia sitophila (formerly Monilia sitophila) 314

Part III Basics of Molecular Methods for Fungal Identification

Introduction 317

Molecular Terminology 318

Overview of Classic Molecular Identification Methods 322

Fungal Targets 322

Selected Current Molecular Methodologies 323

Amplification and Non-Sequencing-Based Identification Methods 323

PCR (Polymerase Chain Reaction) 323

Nested PCR 324

Real-time PCR 324

Melting curve analysis 324

Fluorescence resonance energy transfer (FRET) 325

TaqMan 5′ nuclease 325

Molecular beacons 325

Microarray 326

Repetitive-element PCR (rep-PCR) 327

Sequencing-Based Identification Methods 327

Sanger sequencing 327

Pyrosequencing 328

DNA barcoding 328

Applications of DNA Sequencing 329

Accurate method identification 329

Phylogenetic analysis 330

Organism typing 332

Commercial Platforms and Recently Developed Techniques 332

AccuProbe test 332

PNA FISH 332

Luminex xMAP 333

MALDI-TOF 333

Selected References for Further Information 334

Part IV Laboratory Technique

Laboratory Procedures 339

Collection and Preparation of Specimens 341

Methods for Direct Microscopic Examination of Specimens 344

Primary Isolation 346

Table 30 Media for primary isolation of fungi 348

Table 31 Inhibitory mould agar versus Sabouraud dextrose agar as a primary medium for isolation of fungi 349

Macroscopic Examination of Cultures 349

Microscopic Examination of Growth 350

Procedure for Identification of Yeasts 352

Direct Identification of Yeasts from Blood Culture (by PNA FISH) 354

Isolation of Yeast When Mixed with Bacteria 355

Germ Tube Test for the Presumptive Identification of Candida albicans 356

Rapid Enzyme Tests for the Presumptive Identification of Candida albicans 356

Caffeic Acid Disk Test 357

Olive Oil Disks for Culturing Malassezia species 357

Conversion of Thermally Dimorphic Fungi in Culture 358

Method of Inducing Sporulation of Apophysomyces elegans and Saksenaea vasiformis 358

In Vitro Hair Perforation Test (for Differentiation of Trichophyton mentagrophytes and Trichophyton rubrum) 359

Germ Tube Test for Differentiation of Some Dematiaceous Fungi 359

Temperature Tolerance Testing 360

Maintenance of Stock Fungal Cultures 360

Controlling Mites 361

Staining Methods 363

Acid-Fast Modified Kinyoun Stain for Nocardia spp. 365

Acid-Fast Stain for Ascospores 366

Ascospore Stain 366

Calcofluor White Stain 366

Giemsa Stain 367

Gomori Methenamine Silver (GMS) Stain 368

Gram Stain (Hucker Modification) 370

Lactophenol Cotton Blue 371

Lactophenol Cotton Blue with Polyvinyl Alcohol (PVA) (Huber’s PVA Mounting Medium, Modified) 371

Rehydration of Paraffin-Embedded Tissue (Deparaffination) 372

Media 373

Introduction 375

Acetamide Agar 375

Arylsulfatase Broth 376

Ascospore Media 376

Assimilation Media (for Yeasts) 377

Birdseed Agar (Niger Seed Agar; Staib Agar) 381

Brain Heart Infusion (BHI) Agar 382

Buffered Charcoal Yeast Extract (BCYE) Agar 382

Canavanine Glycine Bromothymol Blue (CGB) Agar 383

Casein Agar 384

CHROMagar Candida Medium 384

ChromID Candida Medium 385

Citrate Agar 386

Cornmeal Agar 386

Dermatophyte Test Medium (DTM) 387

Dixon Agar (Modified) 388

Esculin Agar 388

Fermentation Broths for Yeasts 389

Gelatin Medium 389

Inhibitory Mould Agar (IMA) 391

Leeming-Notman Agar (Modified) 391

Loeffler Medium 392

Lysozyme Medium 392

Middlebrook Agar Opacity Test for Nocardia farcinica 393

Mycosel Agar 393

Nitrate Broth 394

Polished Rice, or Rice Grain, Medium 394

Potato Dextrose Agar and Potato Flake Agar 395

Rapid Assimilation of Trehalose (RAT) Broth 395

Rapid Sporulation Medium (RSM) 397

SABHI Agar 397

Sabouraud Dextrose Agar (SDA) 398

Sabouraud Dextrose Agar with 15% NaCl 399

Sabouraud Dextrose Broth 399

Starch Hydrolysis Agar 399

Trichophyton Agars 400

Tyrosine, Xanthine, or Hypoxanthine Agar 401

Urea Agar 401

Water Agar 402

Yeast Extract-Phosphate Agar with Ammonia 402

Color Plates 405

Glossary 435

Bibliography 447

Selected Websites 465

Index 469

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