CZECHOSLOVAK ACADEMY OF SCIENCES MYCOLOGICAL DIAGNOSIS OF ANIMAL DERMATOPHYTOSES
CZECHOSLOVAK ACADEMY OF SCIENCES SCIENTIFIC EDITOR PROF. DR. KAREL CEJP SCIENTIFIC ADVISER DOC. DR. JIRl MANYCH LANGUAGE EDITOR EVA KALINOV A GRAPHIC DESIGN BY MIROSLAV HOUSKA
MYCOLOGICAL DIAGNOSIS OF ANIMAL DERMATOPHYTOSES JAROSLAV DVORAK MILOS OTCENASEK SPRINGER-SCIENCE+BUSINESS MEDIA, B.V ACADEMIA, PUBLISHING HOUSE OF THE CZECHOSLOVAK ACADEMY OF SCIENCES, PRAGUE
DISTRIBUTION THROUGHOUT THE WORLD WITH THE EXCEPTION OF SOCIALIST COUNTRIES: DH. W. JUNK N. V., PUBLISHERS, 13, VAN STOLKWEG THE HAGUE, HOLLAND ISBN 978-90-6193-236-9 ISBN 978-94-010-3426-5 (ebook) DOI 10.1007/978-94-010-3426-5 SPRINGER SCIENCE+BUSINESS MEDIA DORDRECHT 1969 Originally published by Dr. W. Junk B. V. Publishers 1969 TRANSLATION EVA KALINOVA 1969
DEDICATED TO PROF. Dr. RAYMOND VANBREUSEGHEM PRESIDENT OF THE INTERNATIONAL SOCIETY FOR HUMAN AND ANIMAL MYCOLOGY
CONTENTS List of figures........ List of photographs in the text. List of tables... Preface... Acknowledgements. 10 11 12 13 14 A. General Introduction. I. Historical notes..... II. The term dermatophytosis III. The taxonomy of dermatophytes. IV. The importance of individual dermatophytes in veterinary medicine. V. The location of human dermatophytic lesions...... VI. The propagation of dermatophytes under natural conditions VII. Geographical distribution of dermatophytes VIII. The present-day concept of dermatophytes IX. Diagnostic characters of dermatophytes. X.. Direct microscopy...... XI. Cultivation......... a) Macromorphology of colonies 1. Rate of growth. 2. Ground plan 3. Texture... 4. Surface colour. 5. Reverse colour. 6. Pleomorphic degeneration. 7. Faviform degeneration. b) Micromorphology of colonies 8. Macroconidia..... 9. Microconidia...... 10. Modes of conidial production. 11. Germination of conidia and mycelium formation XII. Other methods.... 1. Differential diagnosis.... 2. Animal inoculation..... 3. Keratinolysis of hairs in vitro 4. Nutritional tests 5. Perfect states....... 19 22 22 23 26 26 27 31 31 31 31 35 36 36 37 37 38 38 38 39 39 42 44 44 45 54 54 55 56 57 57 7
B. Special aspects Zoophilic dermatophytes rarely attacking man (Trichophyton equinum, Trichophyton gallinae)............ 61 Zoophilic dermatophytes commonly attacking man (Microsporon canis, Trichophyton mentagrophytes, Trichophyton?Jerrucosum) 69 Anthropophilic dermatophytes occasionally attacking animals (Epidermophyton floccosum, Microsporon audouinii, Trichophyton megninii, Trichophyton rubrum, Trichophyton schoenleinii, Trichophyton ton8urans, Trichophyton violaceum)....................... 85 Geophilic dermatophytes attacking or infecting animals and man (Keratinomyces ajelloi, Microsporon cookei, Microsporon fulvumfgypseum, Microsporon nanum, 1'richophyton terrestre).............. 102 Dermatophytes rarely isolated from animals and man (Microsporon distortum, Microsporon vanbreuseghemii, Trichophyton simii) 117 Anthropophilic dermatophytes not attacking animals (Microsporon jerrugineum, Trichophyton concentricum, Trichophyton gourvilii, Trichophyton. sudanense, Trichophyton yaoundei)............ 125 Dermatophytes of doubtful systematic position (Microsporon equinum, Sabouraudites d uboisii, Sabouraudites langeronii, Sabouraudites praecox, Trichophyton persicolor, Trichophyton rodhainii, Trichophyton sulphureum)......................... 136 Newly described dermatophytes (Keratinomyceslongijusus, Microsporon rivalierii, Thallomicrosporon kuehnii, Trichophyton georgii, Trichophyton vanbreuseghemii)........... 139 C. Techniques I. Equipment for sample collection II. Collecting material for examination.. III. Microscopical examination of specimens IV. Cultivation........ 1. General instructions... 2. Media for prin1ary isolation 3. Isolation...... 4. Media for subcultures... 5. Special media...... 6. Cultivation in increased C0 2 tension 7. Isolation of colonies... V. Macroscopic examination of cultures VI. Microscopic examination of cultures 1. Preparations in mounting fluids 2. Slide cultures.... VII. Other diagnostic methods.... 1. Nutritional tests....... 2. Study of the manner of growth in liquid media 3. Hair perforation test............ 145 146 148 150 150 151 153 153 154 155 156 156 157 157 158 159 159 161 161 8
VIII. Experimental inoculation of animals.... 161 I. Inoculation of culture......... 162 2. Inoculation of a sample of clinical material 162 IX. Induction of perfect states........ 163 X. Resistance of dermatophyte cultures in mycological collection. 163 XI. Storage of dermatophytic culture in mycological collection 164 I. Maintenance of viable cultures... 164 2. Preservation of dead cultures... 165 3. Benedek's mycotheca in lactophenol 165 XII. Infestation of cultures by mit.es 166 XIII. Recommended procedure for the collection and preservation of important data obtained by laboratory examination 166 Addendum 171 Literature....... 196 Glossary........ 207 Index oo the names of species and varieties 211 9
LIST OF FIGURES Fig. I. Life cycle of dermatophytes under natural conditions. 2. Modes of propagation of geophilic dermatophytes. 3. Modes of propagation of zoophilic dermatophytes commonly infecting man. 4. Modes of propagation of zoophilic dermatophytes occasionally infecting man. 5. Modes of propagat-ion of anthropophilic dermatophytes occasionally infecting man. 6. Dermatophytic elements in skin scrapings. 7. Modes of hair invasion in vivo (formerly used for the. separation of dermatophytes). 8. Approximate diameter (in mm) of colonies after ten days. 9. Shape and structure of macroconidia observed. 10. Number of macroconidial "cells". 11. Modes of macroconidial formation in M. canis. 12. Modes of microconidia! formation. 13. Modes of macroconidial and microconidia! germination. 14. Special hypha! formations. 15. Spores of the vegetative mycelium. 16. Differential diagnosis. 17. Modes of hair keratinolysis in vitro. 18. Perfect states of Arthroderma and Nannizzia, peridial hyphae and appendages. 19. Trichophyton equinum. 20. Trichophyton gallinae. 21. Microsporon canis. 22. Trichophyton mentagrophytes. 23. Trichophyton verrucusum. 24. Epidermophyton floccosum. 25. Microsporon audouinii. 26. Trichophyton megninii. 27. Trichophyton rubrum. 28. Trichophyton schoenleinii. 29. Trichophyton tonsurans. 30. Trichophyton violaceum. 31. Keratinomyces ajelloi. 32. Microsporon cookei. 33. Microsporon gypseum. 34. Micro8'poron nanum. 35. Trichophyton terrestre. 36. Microsporon distortum. 37. Microsporon vanbreuseghemii. 38. Trichophyton B'imii. 39. Microsporon jerrugineum. 40. Trichophyton concentricum. 41. Trichophyton gourvilii. 42. Trichophyton sudanense. 43. Trichophyton yaoundei. 10
LIST OF PHOTOGRAPHS IN THE TEXT l. Dermatophytic mycelium ~n skin scales, KOH preparation (approx. X 200). 2. Cattle hair invaded by Trichophyton verrucosum; KOH preparation (approx. X 500). 3. Mycelium oft. violaceum, iodine solution after 10 min (approx. X 250). 4. Mycelium with macroconidia of JYI. gypseum; slide culture (approx. X 100). 5. Young macroconidia of JYI. gypseum; iodine solution after 10 min (approx. X 750). 6. Mature macroconidium of JYI. gypseum; iodine solution after 10 min (approx. X 200). 7. Mycelium with macroconidia of K. ajelloi; slide culture (approx. X 275). 8. Mature macroconidium of K. ajelloi; iodine solution after 10 min (X 2,000). 9. Sessile macroconidia of E. floccosum; iodine solution after 10 min (X 1,500). 10. Microconidia of T. tonsurans; iodine solution after 10 min (approx. X 250). 11. Chlamydospores oft. tonsurans; iodine solution after 10 min (approx. X 250). 12. Spirals of T. mentagrophytes; iodine solution after 10 min (approx. X 1,000, approx. x 300). 13. Favic chandeliers of T. verrucosum; iodine solution after 10 min (approx. X 500). 11
LIST OF TABLES Table l. Comparison of three classification systems of dermatophytes. 2. Survey of hosts of dermatophytes. 3. Location of human dermatophytic lesions. 4. Geographical distribution of dermatophytes. 5. Anthropophilic, geophilic and zoophilic dermatophytes and their geographical distribution. 6. Fluorescence of invaded hairs under the Wood's light. 7. Modes of hair invasion by dermatophytes in vivo. 8. Ground plan of dermatophyte colonies. 9. Texture of dermatophyte colonies. 10. The tendency to form furrows and folds. 11. Surface colour. 12. Reverse colour. 13. Number of macroconidia. 14. Shape of macroconidia. 15. Size of macroconidia. 16. Number of microconidia. 17. Experimental pathogenicity for guinea-pigs. 18, Supplementary methods of identification of dermatophytes according to the mode of hair keratinolysis in various animals and man. 19. Nutritional tests. 20. Survey of perfect and corresponding imperfect states of dermatophytes. 21. Survey of important features of perfect states. 12
PREFACE There are not many biological disciplines, which have experienced such fast development as have human and veterinary mycology in recent years. In the initial stages organisms, whoee influence on man has not b~en determined, have been investigated on a purely academic basis and with particular reference to their taxonomic position. As recently as the thirties, the pathogenic agents of dermatophytoses were known collectively under such names as "Microsporum" and "Trichophyton". Such designation occurred in most medical and veterinary literature irrespective of the fact that fungi are organisms of a highly complicated and most variable morphological structure, an intricate development and a distribution over a diversity of environments, infecting hosts of the animal kingdom. The greater the credit, therefore, that can be attributed to the pioneers of mycological research, the founders of modern mycology such as R. Sabouraud, M. Langeron, R. Vanbreuseghem, Ch. W. Emmons, who elucidated the importance of mycology in human and veterinary medicine. To one of them, Professor R. V anbreuseghem, this book has been dedicated. In spite of these tremendous efforts, many aspects of the biology and ecology of fungi have still remained obscure in view of the complicated development throughout the various phases, the variability of species and the large numbers of purely saprophytic species occurring in nature. At the present, mycoparasitic infection in man and animals is of utmost importance. Since modern therapeutics have reduced the incidence of various bacterioses and rickettsioses and some viral diseases have been controlled by successful vaccination, increased interest has been taken in fungal diseases. Even the highest medical and veterinary organisations of the UNO focussed their attention on these problems. At the meeting of the WHO and F AO Committee of Experts on Zoonoses, held at the end of 1966 in Geneva, the importance of these problems was stressed by the demand for further research in the field of dermatophytoses and systemic mycoses. This shows the great need of intensified research in medical and veterinary mycology, conducted at various levels. 13
For these reasons the book "Mycological diagnosis of animal dermatophytoses", presented by J. Dvorak and M. Otcenasek of the Institute of Parasitology, Czechoslovak Academy of Sciences, Prague and concerned with the var~ous aspects of animal dermatophytoses, should be most welcomed. The authors have laid down solid foundations for the mycological diagnosis of dermatophytoses in veterinary practice, a problem until now very little discussed in world literature. They present fundamental knowledge on the morphology, development and ecology of fungi, collected over a period of more than 15 years of active work and study. Their experiences in human mycology have been applied successfully in veterinary mycology. The book is divided into three parts. Part One is an introduction to the study of dermatophytes as animal parasites, giving information on standard methods of cultivation and identification. Part Two reviews the species infecting animals; Part Three discusses techniques, which should be used for the exact identification of dermatophytes. The authors have tried to give an accurate picture of fungi that cause infection in animals, by reference to all aspects of contemporary taxonomy, biology and ecology. By recording the results obtained in mycoparasitology of dermatophytes in Czechoslovakia and the other socialist countries and confronting them with the most important achievements in the capitalist countries, the book may be of valuable help by presenting a complete picture on the present sta.te of mycoparasitological research in the world. Mter World War II mycology, following a general world trend, started to be developed in Czechoslovakia. This was done on several levels. In view of the general lay-out of this book, I shall outline this development principally with regard to medical and veterinary mycology. The first systematic study concerned with human dermatophytology, based on 115 isolations of dermatophytes from man during the years 1933 to 1936 and on their detailed study, was published by 0BRTEL (Prague 1936) under the title: "Les caracteristiques morphologiques et biologiques des teignes de la peau se presentant a Prague". Even today, this book has retained its basic value. The Atlas of Dermatophytes, published by the same author in 1950, offers an excellent aid for the diagnosis of these fungi. In 1946, CHMEL recorded a list of dermatophytes isolated from human lesions in 1942-1946. His recent book on dermatophytoses of man and their treatment (CHMEL 1964) also deserves to be mentioned. FRAGNER (1958) published a most useful monograph on this subject, reviewing his longlasting experiences in the laboratory. A more recent publication by HuBSCHMANN and FRAGNER (1962) ondermatophytes and skin diseases caused by them, also merits attention. Shortly after World War II, mycoparasitological laboratories were set up in Prague (Manych- Laboratory for Medical Mycology) and Olomouc (Hejt- 14
manek, Hejtmankova- Biological Institute of the Faculty of Medicine). In 194 7, research on dermatophytes was started at the Dermatophytological Laboratory of the Faculty of Medicine at Hradec Kralove under the leadership of Prof. Fingerland and Prof. Janousek, later under Prof. Malek. As it would be most difficult to name all prominent Czech and Slovak mycologists I shall record only those, whose work has inspired the senior author of this book, Dr. Dvorak, to study medical mycology, particularly mycoparasitology of homoiothermous animals. In the first place the excellent publication by CoRDA (1837-1854), custodian of the National Museum in Prague, entitled: "leones fungorum hucusque cognitorum" and, in recent years "Houby" (Fungi) by CEJP (1957), an important textbook on parasitic fungi and fungi in general, now recognized as one of the fundaments of mycological research in Czechoslovakia. Dr. Dvorak started mycological work at the Dermatophytological Laboratory of the Faculty of Medicine at Hradec Kralove in 1949. In 1954, he published with Docekal the "Manual of Medical Mycology" (DocEKAL and Dvo:RAK 1954). Some years later, this laboratory concerned with the diagnosis of dermatophytoses in eastern Bohemia, was moved to Pardubice. In 1962 the mycological group of the Institute of Parasitology of the Czechoslovak Academy of Sciences was established and there, the author continued in his work in close collaboration with the second author of this book, Dr. Otcenasek. The interest of both authors in animal dermatophytes was fostered by Prof. Sova and Dr. Komarek of the Veterinary Hospital at Pardubice who gave them the opportunity to examine animals, especially cattle and horses, on a more extensive scale. Later they were asked to take over mycological control of large breeding stocks of laboratory animals at Rosice njlabem (Research Institute for Pharmacy and Biochemistry, Director Dr. Hradil). Dr. Dvorak and Dr. Oteenasek, having devoted years to the study of dermatophytes, are now presenting the results of their patient and longlasting work for the benefit of all those, who may use their book in practice to add to their efficiency and economy of time. It is hardly necessary to emphasize that every biologist, regardless of his specialisation, should know the organism with which he is working. Such knowledge is expected ever more from veterinarians and physicians, especially, if the pathogenic agent causes infection in man and animals. The excellent arrangement of the book allows every user to identify the taxonomic position, even as far as the species, of every dermatophyte isolated from any domestic animal. Prof. Dr. B. Rosicky, DSc. Corresponding Member of the Czechoslovak Academy of Sciences Director of the Institute of Parasitology, Prague 15
Acknowledgments We are very indebted to Dr. J. Komarek of the Veterinary Hospital at Pardubice for the list of clinical characteristics of dermatophytoses. All photographs were contributed by Dr. Z. Hubalek, a member of our team, to whom we are happy to acknowledge our indebtedness. We also wish to thank Dr. J. Kunert from the Biological Institute of the Faculty of Medicine, Palacky University, Olomouc for his valuable advice and help in the elaboration of the part concerned with perfect states of dermatophytes. For valuable technical assistance special thanks are given to Mrs. E. Blehova, H. Dyntarova, E. Kalinova, M. Kralova and J. Kuchafova from the Institute of Parasitology, Czechoslovak Academy of Sciences, Prague, to V. Jaromersky and A. Licbinska from the District Station of Hygiene and Epidemiology, Pardubice and to Dr. J. Holda, Institute for Pharmacy and Biochemistry, Rosice n/labem. For the provision or identification of dermatophytes and Chrysosporia and other valuable information, we express our thank~ to: V. A. Balabanoff (Sofia), J. Buchvald (Bratislava), J. M. Doby (Rennes), Mrs. M. Doby Dubois (Rennes), R. Evolceanu (Bucharest), E. Florian (Budapest), D. Frey (Sydney), L. K. Georg (Atlanta), D. Hantschke (Essen), M. Hejtmanek (Olomouc), N. Hejtmankova (Olomouc), Z. Herpay (Debrecen), J. Manych (Prague), L. Ozegovic (Sarajevo), H. Paldrok (Stockholm), J. A. Rioux (Mont. pellier), Ch. Schonborn (Leipzig), R. Vanbreuseghem (Antwerp). 16