Larone’s MEDICALLY IMPORTANT FUNGI The definitive guide for identifying fungi from clinical specimens
With a new team of authors, Larone’s Medically Important Fungi, Seventh Edition, continues the longstanding tradition of high-quality content to expand your knowledge and support your work in clinical mycology 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 more than 150 of the fungi most commonly encountered in the clinical mycology laboratory, including new entries for Emergomyces, Metarhizium anisopliae, Rasamsonia argillacea, Rhinocladiella mackenziei, Schizophyllum commune, and Thermothelomyces thermophilus Presenting details on each organism’s pathogenicity, growth characteristics, relevant biochemical reactions, and microscopic morphology, illustrated with photomicrographs, unique and elegant drawings, and color photos of colony morphology and various test results Explaining 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., nucleic acid amplification and 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 confidently identify commonly encountered fungi.
List of Tables xvii Preface to the Seventh Edition xix Preface to the First Edition xxi Acknowledgments xxiii About the Authors xxv Basics 1 How To Use the Guide 3 Use of Reference Laboratories and Regulations for Transport 5 Safety Precautions 11 Taxonomy and Nomenclature 13 Part I Direct Microscopic Examination of Clinical Specimens 15 Introduction 17 Histological Terminology 21 Tissue Reactions to Fungal Infection 25 Stains 29 Table 1.1 Histochemical stains for fungi and/or filamentous bacteria in tissue 30 Guide to Interpretation of Direct Microscopic Examination 33 Detailed Descriptions 39 Actinomycosis 40 Mycetoma (Actinomycotic or Eumycotic) 41 Nocardiosis 43 Mucormycosis (Zygomycosis) 44 Aspergillosis 45 Miscellaneous Hyalohyphomycoses (Other than Aspergillosis) 47 Dermatophytosis (Tinea, Ringworm) 49 Tinea versicolor 50 Tinea nigra 51 Phaeohyphomycosis 52 Chromoblastomycosis 53 Sporotrichosis 54 Histoplasmosis 55 Emergomycosis 57 Talaromycosis (Penicilliosis) 58 Blastomycosis 59 Paracoccidioidomycosis 60 Lobomycosis 61 Candidiasis 62 Trichosporonosis 64 Cryptococcosis 65 Pneumocystosis 67 Protothecosis 68 Coccidioidomycosis 69 Rhinosporidiosis 70 Adiaspiromycosis 72 Part II Identification of Fungi in Culture 73 Guide to Identification of Fungi in Culture 75 Detailed Descriptions 105 Filamentous Bacteria 107 Introduction to Filamentous Bacteria 109 Table 2.1 Differentiation of filamentous aerobic actinomycetes encountered in clinical specimens 110 Nocardia spp. 111 Streptomyces spp. 114 Actinomadura spp. 116 Nocardiopsis dassonvillei 117 Yeasts and Yeastlike Organisms 119 Introduction to Yeasts and Yeastlike Organisms 121 Candida albicans 123 Table 2.2 Characteristics of the genera of clinically encountered yeasts and yeastlike organisms 124 Candida dubliniensis 125 Table 2.3 Characteristics of Candida spp. most commonly encountered in the clinical laboratory 126 Table 2.4 Characteristics that assist in differentiating Candida dubliniensis from Candida albicans 128 Candida tropicalis 129 Candida parapsilosis species complex 130 Candida lusitaniae 131 Candida krusei 132 Table 2.5 Differentiating characteristics of Magnusiomyces capitatus (formerly Blastoschizomyces capitatus) versus Candida krusei 134 Table 2.6 Differentiating characteristics of Candida krusei, Candida inconspicua, and Candida norvegensis 134 Candida kefyr 135 Candida rugosa species complex 136 Candida guilliermondii species complex 138 Table 2.7 Differentiating characteristics of Candida guilliermondii versus Candida famata 139 Candida lipolytica 140 Candida zeylanoides 141 Candida glabrata species complex 142 Candida auris 143 Candida haemulonii species complex 144 Candida pelliculosa 145 Cryptococcus neoformans species complex 147 Cryptococcus gattii species complex 149 Table 2.8 Characteristics of Cryptococcus spp. and former members of the genus 150 Table 2.9 Characteristics of yeasts and yeastlike organisms other than Candida spp. and Cryptococcus spp. 151 Rhodotorula and Cystobasidium spp. 152 Sporobolomyces salmonicolor 154 Saccharomyces cerevisiae 156 Malassezia spp. 158 Malassezia pachydermatis 160 Ustilago spp. 161 Prototheca spp. 162 Trichosporon and Cutaneotrichosporon spp. 163 Table 2.10 Key characteristics of the most common clinically encountered Trichosporon spp. and Cutaneotrichosporon spp. 165 Magnusiomyces capitatus (formerly Blastoschizomyces capitatus) 166 Geotrichum candidum 167 Thermally Dimorphic and/or Endemic Fungi 169 Introduction to Thermally Dimorphic and/or Endemic Fungi 171 Histoplasma capsulatum 172 Emergomyces spp. 175 Blastomyces dermatitidis/gilchristii 177 Coccidioides immitis/posadasii 179 Paracoccidioides brasiliensis 181 Talaromyces marneffei (formerly Penicillium marneffei) 183 Sporothrix schenckii species complex 186 Emmonsia crescens 189 Thermally Monomorphic Moulds 191 Mucormycetes 193 Introduction to Mucormycetes 195 Table 2.11 Differential characteristics of similar organisms in the class Mucormycetes 197 Table 2.12 Differential characteristics of the clinically encountered Rhizopus spp. 197 Rhizopus spp. 198 Mucor spp. 200 Rhizomucor spp. 201 Lichtheimia corymbifera species complex 202 Apophysomyces elegans species complex 204 Saksenaea vasiformis 206 Cokeromyces recurvatus 207 Cunninghamella bertholletiae 209 Syncephalastrum racemosum 211 Basidiobolus spp. 212 Conidiobolus coronatus 213 Dematiaceous Fungi 215 Introduction to Dematiaceous Fungi 217 Fonsecaea spp. 218 Myrmecridium schulzeri 221 Rhinocladiella mackenziei (formerly Ramichloridium mackenziei) 222 Phialophora verrucosa 223 Table 2.13 Characteristics of Phialophora, Pleurostoma (formerly Pleurostomophora), Phaeoacremonium, Acremonium and Sarocladium, Phialemonium, and Coniochaeta (formerly Lecythophora) 224 Pleurostoma richardsiae (formerly Pleurostomophora richardsiae) 225 Phaeoacremonium parasiticum 226 Phialemonium spp. 228 Cladosporium spp. 230 Table 2.14 Characteristics of Cladosporium spp. and Cladophialophora spp. 232 Cladophialophora carrionii 233 Cladophialophora boppii 235 Cladophialophora bantiana 236 Scedosporium apiospermum species complex 237 Table 2.15 Differentiating phenotypic characteristics of the clinically encountered members of Scedosporium spp. and Lomentospora prolificans 241 Lomentospora prolificans (formerly Scedosporium prolificans) 242 Verruconis gallopava (formerly Ochroconis gallopava) 244 Table 2.16 Characteristics of some of the “black yeasts” 246 Exophiala jeanselmei species complex 247 Exophiala dermatitidis 249 Hortaea werneckii 251 Madurella mycetomatis 252 Trematosphaeria grisea (formerly Madurella grisea) 253 Piedraia hortae 254 Aureobasidium pullulans 255 Table 2.17 Differential characteristics of Aureobasidium pullulans versus Hormonema dematioides 257 Hormonema dematioides 258 Neoscytalidium dimidiatum 259 Botrytis cinerea 261 Stachybotrys chartarum 262 Thermothelomyces thermophilus (formerly Myceliophthora thermophila) 264 Curvularia spp. 265 Table 2.18 Characteristics of Curvularia spp. and Exserohilum rostratum 269 Exserohilum rostratum 270 Helminthosporium spp. 272 Alternaria spp. 273 Stemphylium spp. 275 Pseudopithomyces spp. (formerly Pithomyces spp.) 276 Epicoccum spp. 277 Nigrospora spp. 279 Chaetomium spp. 280 Phoma spp. 282 Dermatophytes 285 Introduction to Dermatophytes 287 Latin Terms for Dermatophyte Infections 288 Microsporum audouinii 289 Microsporum canis 290 Paraphyton cookei species complex (formerly Microsporum cookei species complex) 292 Nannizzia gypsea species complex (formerly Microsporum gypseum species complex) 293 Lophophyton gallinae (formerly Microsporum gallinae [zoophilic form] and Microsporum vanbreuseghemii [geophilic form]) 295 Nannizzia nana (formerly Microsporum nanum) 297 Microsporum ferrugineum 299 Trichophyton mentagrophytes species complex 300 Table 2.19 Differentiation of similar conidia-producing Trichophyton spp. and Arthroderma spp. 302 Trichophyton rubrum 303 Trichophyton tonsurans 305 Arthroderma terrestre species complex (formerly Trichophyton terrestre species complex) 307 Trichophyton megninii 308 Trichophyton soudanense 309 Table 2.20 Growth patterns of Trichophyton spp. and Arthroderma spp. on nutritional test media 310 Trichophyton schoenleinii 311 Trichophyton verrucosum 312 Trichophyton violaceum 313 Arthroderma uncinatum (formerly Trichophyton ajelloi) 314 Epidermophyton floccosum 315 Hyaline Hyphomycetes 317 Introduction to Hyaline Hyphomycetes 319 Fungi in Which Arthroconidia Predominate Table 2.21 Differential characteristics of fungi in which arthroconidia predominate 320 Malbranchea spp. 321 Pseudogymnoascus pannorum (formerly Geomyces pannorum) 323 Arthrographis kalrae 324 Hormographiella aspergillata 326 Common Species of Aspergillus The genus Aspergillus 327 Table 2.22 Differentiating characteristics of the most common Aspergillus spp. 329 Aspergillus fumigatus species complex 331 Aspergillus niger species complex 333 Aspergillus flavus species complex 334 Aspergillus versicolor species complex 336 Aspergillus ustus species complex 338 Aspergillus tanneri 340 Aspergillus nidulans species complex 342 Aspergillus glaucus 344 Aspergillus terreus species complex 345 Aspergillus clavatus 347 Other Common Hyaline Hyphomycetes Penicillium spp. 348 Paecilomyces variotii 350 Rasamsonia argillacea species complex (formerly Geosmithia argillacea) 351 Purpureocillium lilacinum (formerly Paecilomyces lilacinus) 352 Table 2.23 Differential characteristics of Paecilomyces variotii, Rasamsonia argillacea, and Purpureocillium lilacinum 354 Scopulariopsis spp. 355 Table 2.24 Differential characteristics of Scopulariopsis brevicaulis versus Scopulariopsis brumptii 357 Gliocladium spp. 358 Trichoderma spp. 359 Metarhizium anisopliae species complex 361 Beauveria bassiana 363 Verticillium spp. 364 Acremonium and Sarocladium spp. 365 Fusarium spp. 367 Coniochaeta spp. (formerly Lecythophora spp.) 369 Trichothecium roseum 370 Chrysosporium spp. 371 Table 2.25 Differential characteristics of Chrysosporium versus Sporotrichum 373 Sporotrichum pruinosum 374 Sepedonium spp. 376 Chrysonilia sitophila 377 Schizophyllum commune 378 Part III Basics of Molecular Methods for Fungal Identification 379 Introduction 381 Fungal Targets 383 Table 3.1 Frequently used fungal molecular targets and primers for sequence-based species identification 385 Table 3.2 Examples of fungal molecular targets and primers for multilocus sequence-based species identification 386 Classic Molecular Identification Methods 387 Polymerase Chain Reaction 387 Non-Sequencing-Based Identification Methods 389 MALDI-TOF Mass Spectrometry 389 Signal Amplification Methods 390 PNA FISH 390 Nucleic Acid Amplification Methods 390 T2 Magnetic Resonance 390 Broad-Panel Molecular Testing and Other Emerging Sample-to-Answer Technologies 391 Sequencing-Based Identification Methods 393 Sanger Sequencing 393 Table 3.3 Lane construction for traditional bidirectional Sanger sequencing 394 Massive Parallel or Next-Generation Sequencing 394 Applications of DNA Sequencing 397 Accurate Molecular Identification 397 Table 3.4 Commonly used databases for identification of medically important fungi 398 Phylogenetic Analysis 399 Organism Typing 401 Detection of Genetic Determinants of Resistance 401 Part IV Laboratory Technique 403 Laboratory Procedures 405 Collection and Preparation of Specimens 406 Table 4.1 Common clinical sites for laboratory recovery of pathogenic fungi 409 Methods for Direct Microscopic Examination of Specimens 414 Primary Isolation 416 Table 4.2 Media for primary isolation of fungi 417 Macroscopic Examination of Cultures 419 Microscopic Examination of Growth 419 Procedure for Identification of Yeasts 421 Isolation of Yeast When Mixed with Bacteria 424 Germ Tube Test for the Presumptive Identification of Candida albicans 425 Rapid Enzyme Tests for the Presumptive Identification of Candida albicans 426 Caffeic Acid Disk Test 426 Olive Oil Disks for Culturing Malassezia spp. 426 Conversion of Thermally Dimorphic Fungi in Culture 427 Sporulation Inducement Method for Apophysomyces and Saksenaea 428 In vitro Hair Perforation Test (for Differentiation of Trichophyton mentagrophytes and Trichophyton rubrum) 428 Temperature Tolerance Testing 429 Maintenance of Stock Fungal Cultures 429 Controlling Mites 430 Staining Methods 431 Acid-Fast Modified Kinyoun Stain for Nocardia spp. 432 Acid-Fast Stain for Ascospores 433 Ascospore Stain 433 Calcofluor White Stain 434 Giemsa Stain 435 Gomori Methenamine Silver (GMS) Stain 435 Gram Stain (Hucker Modification) 438 Lactophenol Cotton Blue 439 Lactophenol Cotton Blue with Polyvinyl Alcohol (PVA) (Huber’s PVA Mounting Medium, Modified) 439 Rehydration of Paraffin-Embedded Tissue (Deparaffination) 440 Media 441 Ascospore Media 443 Assimilation Media (for Yeasts) 444 Birdseed Agar (Niger Seed Agar; Staib Agar) 448 Brain Heart Infusion (BHI) Agar 449 Buffered Charcoal-Yeast Extract (BCYE) Agar 450 Canavanine Glycine Bromothymol Blue (CGB) Agar 450 Casein Agar 451 CHROMagar Candida Medium 452 CHROMagar Candida Plus Medium 453 CHROMID Candida Agar 453 Chromogenic Candida Agar (Brilliance Candida Agar) 454 Cornmeal Agar 454 Dermatophyte Test Medium (DTM) 455 Esculin Agar 456 Fermentation Broth for Yeasts 456 Inhibitory Mould Agar (IMA) 457 Leeming-Notman Agar (Modified) 458 Lysozyme Medium 458 Mycosel Agar 459 Potato Dextrose Agar and Potato Flake Agar 460 Rapid Assimilation of Trehalose (RAT) Broth 460 Sabouraud Brain Heart Infusion Agar (SABHI Agar) 462 Sabouraud Dextrose Agar (SDA) 463 Sabouraud Dextrose Agar with 15% NaCl 464 Sabouraud Dextrose Broth 464 Starch Hydrolysis Agar 464 Trichophyton Agars 465 Tyrosine Agar 466 Urea Agar 467 Water Agar 467 Image Appendix 469 Glossary 477 References Cited 489 Index 507
Lars F. Westblade, PhD, D(ABMM) is the Director of the Clinical Microbiology Service at NewYork-Presbyterian/Weill Cornell Medical Center and Associate Professor at Weill Cornell Medicine with a primary appointment in the Department of Pathology and Laboratory Medicine and a secondary appointment in the Division of Infectious Diseases, Department of Medicine. He earned his doctoral degree from the University of Birmingham in the United Kingdom, and completed a fellowship in medical and public health laboratory microbiology at Washington University School of Medicine in St. Louis. Eileen M. Burd, PhD, D(ABMM) is the Director of the Clinical Microbiology Laboratory at Emory University Hospital and Professor at Emory University School of Medicine with a primary appointment in the Department of Pathology and Laboratory Medicine and a secondary appointment in the Department of Medicine, Division of Infectious Diseases. She earned her doctoral degree from the Medical College of Wisconsin in Milwaukee and was the Division Head of Microbiology at Henry Ford Hospital in Detroit, Michigan prior to joining the faculty at Emory University in 2007. Shawn R. Lockhart, PhD, D(ABBM) FAAM is the Senior Clinical Laboratory Advisor in the Mycotic Diseases Branch at the Centers for Disease Control and Prevention. He earned his doctoral degree from the University of Kentucky and completed his clinical microbiology fellowship at the University of Iowa Hospitals and Clinics. He directs the CDC training course in mold identification. Gary W. Procop, MD, MS is the CEO of the American Board of Pathology and Professor of Pathology at the Cleveland Clinic Lerner School of Medicine. He remains a Consulting Staff for the Institute of Pathology and Laboratory Medicine, where he served as Medical Director for the Mycology Laboratory for more than two decades. He earned his doctoral degree from the Marshall University School of Medicine. His residency in anatomic and clinical pathology was completed at Duke University and his medical microbiology fellowship at the Mayo Clinic.
Reviews for Larone's Medically Important Fungi: A Guide to Identification
""Replete with explanations of basic histological terms and descriptions of fundamental tissue reactions to fungal infection, the book achieves the remarkable feat of being simultaneously concise and complete."" —Emerging Infectious Diseases, Vol. 30 No. 3, March 2024
