Larone’s Medically Important Fungi

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.

The definitive guide for identifying fungi from clinical specimensMedically 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 glossaryWith 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.

List of Tables xvPreface to the Fifth Edition xviiPreface to the First Edition xixAcknowledgments xxiHow to Use the Guide 1Use of Reference Laboratories 3Safety Precautions 7Part I Direct Microscopic Examination of Clinical SpecimensIntroduction 11Histological Terminology 13Tissue Reactions to Fungal Infection 17Stains 21Table 1 Stains for direct microscopic observation of fungi and/or filamentous bacteria in tissue 22Guide to Interpretation of Direct Microscopic Examination 23Detailed Descriptions 31Actinomycosis 33Mycetoma, Actinomycotic or Eumycotic 34Nocardiosis 36Zygomycosis 37Aspergillosis 38Miscellaneous Hyalohyphomycoses 40Dermatophytosis 42Tinea versicolor 43Tinea nigra 44Phaeohyphomycosis 45Chromoblastomycosis 46Sporotrichosis 47Histoplasmosis capsulati 48Penicilliosis marneffei 50Blastomycosis 52Paracoccidioidomycosis 53Candidiasis (Candidosis) 54Trichosporonosis 56Cryptococcosis 57Pneumocystosis 59Protothecosis 60Coccidioidomycosis 61Rhinosporidiosis 62Adiaspiromycosis 64Special References 65Part II Identification of Fungi in CultureGuide to Identification of Fungi in Culture 69Detailed Descriptions 101Filamentous Bacteria 103Introduction 105Table 2 Differentiation of filamentous aerobic actinomycetes encountered in clinical specimens 107Nocardia spp. 108Table 3 Phenotypic characteristics of most common clinically encountered Nocardia spp. 110Streptomyces spp. 111Actinomadura spp. 112Nocardiopsis dassonvillei 113Yeasts and Yeastlike Organisms 115Introduction 117Candida albican 119Table 4 Characteristics of the genera of clinically encountered yeasts and yeastlike organisms 120Candida dubliniensis 121Table 5 Characteristics of Candida spp. most commonly encountered in the clinical laboratory 122Table 6 Characteristics that assist in differentiating Candida dubliniensis from Candida albicans 124Candida tropicalis 125Candida parapsilosis complex 126Candida lusitaniae 127Candida krusei 128Table 7 Differentiating characteristics of Blastoschizomyces capitatus versus Candida krusei 129Table 8 Differentiating characteristics of C. krusei, C. inconspicua, and C. norvegensis 129Candida kefyr (formerly Candida pseudotropicalis) 130Candida rugosa 131Candida guilliermondii complex 132Table 9 Differentiating characteristics of Candida guilliermondii versus Candida famata 133Candida lipolytica 134Candida zeylanoides 135Candida glabrata 136Cryptococcus neoformans 137Cryptococcus gattii 138Table 10 Characteristics of Cryptococcus spp. 139Table 11 Characteristics of yeasts and yeastlike organisms other than Candida spp. and Cryptococcus spp. 140Rhodotorula spp. 141Sporobolomyces salmonicolor 142Saccharomyces cerevisiae 143Wickerhamomyces anomalus (formerly Pichia anomala and Hansenula anomala) (sexual state); Candida pelliculosa (asexual state) 145Malassezia spp. 146Malassezia pachydermatis 148Ustilago sp. 149Prototheca spp. 150Trichosporon spp. 151Table 12 Key characteristics of the most common clinically encountered Trichosporon spp. 152Blastoschizomyces capitatus 153Geotrichum candidum 154Thermally Dimorphic Fungi 155Introduction 157Histoplasma capsulatum 158Blastomyces dermatitidis 160Paracoccidioides brasiliensis 162Penicillium marneffei 164Sporothrix schenckii complex 166Table 13 Characteristics for differentiating species of the Sporothrix schenckii complex 168Thermally Monomorphic Moulds 169Zygomycetes 171Introduction 173Table 14 Differential characteristics of similar organisms in the class Zygomycetes 175Table 15 Differential characteristics of the clinically encountered Rhizopus spp. 175Rhizopus spp. 176Mucor spp. 177Rhizomucor spp. 178Lichtheimia corymbifera complex (formerly Absidia corymbifera) 179Apophysomyces elegans 181Saksenaea vasiformis 183Cokeromyces recurvatus 184Cunninghamella bertholletiae 185Syncephalastrum racemosum 186Basidiobolus sp. 187Conidiobolus coronatus 188Dematiaceous Fungi 189Introduction 191Fonsecaea pedrosoi 193Fonsecaea compacta 195Rhinocladiella spp. 196Phialophora verrucosa 197Table 16 Characteristics of Phialophora, Pleurostomophora, Phaeoacremonium, Acremonium, Phialemonium, and Lecythophora 198Pleurostomophora richardsiae (formerly Phialophora richardsiae) 199Phaeoacremonium parasiticum (formerly Phialophora parasitica) 200Phialemonium spp. 201Cladosporium spp. 203Table 17 Characteristics of Cladosporium and Cladophialophora spp. 204Cladophialophora carrionii 205Cladophialophora bantiana 206Cladophialophora boppii (formerly Taeniolella boppii) 207Pseudallescheria boydii (sexual state) / Scedosporium apiospermum (asexual state) complex 208Table 18 Differentiating phenotypic characteristics of the clinically encountered members of the Pseudallescheria boydii complex and Scedosporium prolificans 210Scedosporium prolificans (formerly Scedosporium inflatum) 211Ochroconis gallopava (formerly Dactylaria constricta var.gallopava) 212Table 19 Differentiation of the clinically encountered Ochroconis species 213Table 20 Characteristics of some of the "black yeasts" 213Exophiala jeanselmei complex 214Exophiala dermatitidis (Wangiella dermatitidis) 215Hortaea werneckii (Phaeoannellomyces werneckii) 216Madurella mycetomatis 217Madurella grisea 218Piedraia hortae 219Aureobasidium pullulans 220Table 21 Differential characteristics of Aureobasidium pullulans versus Hormonema dematioides 222Hormonema dematioides 223Neoscytalidium dimidiatum (formerly Scytalidium dimidiatum) 224Botrytis sp. 226Stachybotrys chartarum (S. alternans, S. atra) 227Graphium eumorphum 228Curvularia spp. 229Bipolaris spp. 230Table 22 Characteristics of Bipolaris, Drechslera, and Exserohilum spp. 231Exserohilum spp. 232Helminthosporium sp 233Alternaria sp 234Ulocladium sp. 235Stemphylium sp. 236Pithomyces sp. 237Epicoccum sp. 238Nigrospora sp. 239Chaetomium sp. 240Phoma spp. 241Dermatophytes 243Introduction 245Microsporum audouinii 247Microsporum canis var. canis 248Microsporum canis var. distortum 249Microsporum cookei 250Microsporum gypseum complex 251Microsporum gallinae 252Microsporum nanum 253Microsporum vanbreuseghemii 254Microsporum ferrugineum 255Trichophyton mentagrophytes 256Table 23 Differentiation of similar conidia-producing Trichophyton spp 257Trichophyton rubrum 258Trichophyton tonsurans 259Trichophyton terrestre 260Trichophyton megninii 261Trichophyton soudanense 262Table 24 Growth patterns of Trichophyton species on nutritional test media 263Trichophyton schoenleinii 264Trichophyton verrucosum 265Trichophyton violaceum 266Trichophyton ajelloi 267Epidermophyton floccosum 268Hyaline Hyphomycetes 269Introduction 271Coccidioides spp. 272Table 25 Differential characteristics of fungi in which arthroconidia predominate 274Malbranchea spp. 275Geomyces pannorum 276Arthrographis kalrae 277Hormographiella aspergillata 278Emmonsia spp. 279The Genus Aspergillus 281Aspergillus fumigatus 283Aspergillus niger 284Aspergillus flavus 285Table 26 Identification of the most common species of Aspergillus 286Aspergillus versicolor 288Aspergillus calidoustus 289Aspergillus nidulans (asexual state); Emericella nidulans (sexual state) 290Aspergillus glaucus (asexual state); Eurotium herbariorum (sexual state) 291Aspergillus terreus 292Aspergillus clavatus 293Penicillium spp. 294Paecilomyces spp. 295Scopulariopsis spp. 297Table 27 Differential characteristics of Paecilomyces variotii versus P. lilacinus 299Table 28 Differential characteristics of Scopulariopsis brevicaulis versus S. brumptii 299Gliocladium sp. 300Trichoderma sp. 301Beauveria bassiana 302Verticillium sp. 303Acremonium (formerly Cephalosporium) spp. 304Fusarium spp. 305Lecythophora spp. 307Trichothecium roseum 308Chrysosporium spp. 309Table 29 Differential characteristics of Chrysosporium versus Sporotrichum 311Sporotrichum pruinosum 312Sepedonium sp. 313Chrysonilia sitophila (formerly Monilia sitophila) 314Part III Basics of Molecular Methods for Fungal IdentificationIntroduction 317Molecular Terminology 318Overview of Classic Molecular Identification Methods 322Fungal Targets 322Selected Current Molecular Methodologies 323Amplification and Non-Sequencing-Based Identification Methods 323PCR (Polymerase Chain Reaction) 323Nested PCR 324Real-time PCR 324Melting curve analysis 324Fluorescence resonance energy transfer (FRET) 325TaqMan 5' nuclease 325Molecular beacons 325Microarray 326Repetitive-element PCR (rep-PCR) 327Sequencing-Based Identification Methods 327Sanger sequencing 327Pyrosequencing 328DNA barcoding 328Applications of DNA Sequencing 329Accurate method identification 329Phylogenetic analysis 330Organism typing 332Commercial Platforms and Recently Developed Techniques 332AccuProbe test 332PNA FISH 332Luminex xMAP 333MALDI-TOF 333Selected References for Further Information 334Part IV Laboratory TechniqueLaboratory Procedures 339Collection and Preparation of Specimens 341Methods for Direct Microscopic Examination of Specimens 344Primary Isolation 346Table 30 Media for primary isolation of fungi 348Table 31 Inhibitory mould agar versus Sabouraud dextrose agar as a primary medium for isolation of fungi 349Macroscopic Examination of Cultures 349Microscopic Examination of Growth 350Procedure for Identification of Yeasts 352Direct Identification of Yeasts from Blood Culture (by PNA FISH) 354Isolation of Yeast When Mixed with Bacteria 355Germ Tube Test for the Presumptive Identification of Candida albicans 356Rapid Enzyme Tests for the Presumptive Identification of Candida albicans 356Caffeic Acid Disk Test 357Olive Oil Disks for Culturing Malassezia species 357Conversion of Thermally Dimorphic Fungi in Culture 358Method of Inducing Sporulation of Apophysomyces elegans and Saksenaea vasiformis 358In Vitro Hair Perforation Test (for Differentiation of Trichophyton mentagrophytes and Trichophyton rubrum) 359Germ Tube Test for Differentiation of Some Dematiaceous Fungi 359Temperature Tolerance Testing 360Maintenance of Stock Fungal Cultures 360Controlling Mites 361Staining Methods 363Acid-Fast Modified Kinyoun Stain for Nocardia spp. 365Acid-Fast Stain for Ascospores 366Ascospore Stain 366Calcofluor White Stain 366Giemsa Stain 367Gomori Methenamine Silver (GMS) Stain 368Gram Stain (Hucker Modification) 370Lactophenol Cotton Blue 371Lactophenol Cotton Blue with Polyvinyl Alcohol (PVA) (Huber's PVA Mounting Medium, Modified) 371Rehydration of Paraffin-Embedded Tissue (Deparaffination) 372Media 373Introduction 375Acetamide Agar 375Arylsulfatase Broth 376Ascospore Media 376Assimilation Media (for Yeasts) 377Birdseed Agar (Niger Seed Agar; Staib Agar) 381Brain Heart Infusion (BHI) Agar 382Buffered Charcoal Yeast Extract (BCYE) Agar 382Canavanine Glycine Bromothymol Blue (CGB) Agar 383Casein Agar 384CHROMagar Candida Medium 384ChromID Candida Medium 385Citrate Agar 386Cornmeal Agar 386Dermatophyte Test Medium (DTM) 387Dixon Agar (Modified) 388Esculin Agar 388Fermentation Broths for Yeasts 389Gelatin Medium 389Inhibitory Mould Agar (IMA) 391Leeming-Notman Agar (Modified) 391Loeffler Medium 392Lysozyme Medium 392Middlebrook Agar Opacity Test for Nocardia farcinica 393Mycosel Agar 393Nitrate Broth 394Polished Rice, or Rice Grain, Medium 394Potato Dextrose Agar and Potato Flake Agar 395Rapid Assimilation of Trehalose (RAT) Broth 395Rapid Sporulation Medium (RSM) 397SABHI Agar 397Sabouraud Dextrose Agar (SDA) 398Sabouraud Dextrose Agar with 15% NaCl 399Sabouraud Dextrose Broth 399Starch Hydrolysis Agar 399Trichophyton Agars 400Tyrosine, Xanthine, or Hypoxanthine Agar 401Urea Agar 401Water Agar 402Yeast Extract-Phosphate Agar with Ammonia 402Color Plates 405Glossary 435Bibliography 447Selected Websites 465Index 469

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