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Growth and Morphology of Piedraia Hortae (Brumpt) Fonseca and a R & Leao: the Causal Agent of Black Piedra

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Growth and Morphology of Piedraia Hortae (Brumpt) Fonseca and a R & Leao: the Causal Agent of Black Piedra I ' i I- 77-2426 JONES, Jeanette, 1950- GROWTH AND MORPHOLOGY OF PIEDRAIA HORTAE (BRUMPT) FONSECA AND A R & LEAO: THE CAUSAL AGENT OF BLACK PIEDRA. The Ohio State University, Ph.D., 1976 Botany Xerox University Microfilms,Ann Arbor, Michigan 48106 © 1976 JEANETTE JONES ALL RIGHTS RESERVED Growth and Morphology of Piedraia hortaa (Brumpt) Fonseca and Area Leao The Causal Agent of Black Piedra Dissertation Presented in Partial Fulfillment of the Requirements for the Degree Doctor of Philosophy in the Graduate School of The Ohio State University hy Jeanette Jones, B. Sc., M. Sc, The Ohio State University 1976 Reading Committee Approved by J. A, Schmitt, Chairman M* 0. Garraway Adviser V, Raghavan Department of Botany R. L. Seymour C. E. Taft ••..the fungi are not degenerate organisms which are on their way out in a scheme of evolution, and so of little economic importance and scientific interest. The fungi, on the contrary, are progressive, ever changing and evolving rapidly in their own way so that they are capable of becoming readily adapted to every condition of life. We may rest assured that as Green Plants and Animals disappear one by one from the face of the globe, some of the fungi will always be present to dispose of the last remains. B. 0. Dodge (1939) ii ACKNOWLEDGEMENTS I am particularly indebted to Dr. J. A Schmitt for his guidance and support when they were needed most. A special thanks is extended to the members of the reading committee, Dr. J. A. Schmitt, Dr. M. 0. Garraway, Dr. V. Raghavan, Dr. C. E. Taft and especially Dr. R. L. Seymour for their critical reading of this manuscript and their help­ ful suggestions during the final wording of the manuscript. Gratitude is also extended to Dr. R. Vanbreuseghem, Institute of Tropical Medicine, Antwerpen, for providing the isolates of Piedraia hortae, and the hair specimens used in this study. Appreciation is extended to the: various individuals in the Depart­ ment of Botany, both faculty and graduate students, for their suggestions and/or technical assistance. Appreciation is also extended to my parents, Willie and Alice Jones, my brothers, and sisters, relatives and friends, who have been under­ standing, thoughtful and encouraging during my graduate studies. Thanks are extended to my undergraduate adviser, Dr. W. D. Moorehead and the many people not mentioned by name who have contributed to making this manuscript possible. iii Vita Sept. 19, 1950 ............... Born, Fort Valley, Georgia 1972 ........... ».B. Sc., The Fort Valley State College, Fort Valley, Georgia 1972 Instructor of Biology Fort Valley State College 1972 ..................................... Research Assistant Southeastern Forestry Exp. Laboratory, Macon, Georgia 1972-73 ........................ ......... University Fellow, Dept, of Botany, The Ohio State University, Columbus, Ohio 1973-75 ................... ...Graduate Teaching Associate The Ohio State University Columbus, Ohio 1975-76 ................... ............ Dissertation Year Fellow The Ohio State University Columbus, Ohio 1976 .......... ................ Consultant, Medical and Other Health Professions Career Day Fort Valley State College Fort Valley, Georgia Publication Jones, Jeanette and J. A. Schmitt, The Effects of Chlorination on the Survival of cells of Candida albicans. Mycologi’a (in press) Field of Study Major field: medical mycology Studies in medical mycology, Professor J. A. Schmitt Studies in general and aquatic mycology, Professor R. L. Seymour Studies in fungal physiology, Professor M. 0. Garraway iv TABLE OF CONTENTS Page ACKNOWLEDGEMENTS 00a0*0*000a00#*a000»0a00a00a000«00a«a0»a0#000 ill VITA 00*0*OMO«OOIOMOOO«OMDOOOOOOMO*OI*ll»OOIOO««»«MOOOtOO IV LIST OF TABLES #00oo oooo»oo*ooooo*o*o**o*o*o*«n**oo«**o**o*o*o VX LIST OF FIGURES ........... viii LIST OF PLATES .... ...c.a...... ix INTRODUCTION «a*aaoo*o*o*»«*eoo»ooo**«**o**a»*oao»»o**»»»os*oo i MATERIALS AND METHODS.................. 4 RESULTS AND DISCUSSION...... .......... .................... Chapter ™ VITRO INVASION OF H A I R ............................ 12 A, Introduction Ba Hyphal Structure C. Hyphal Development on Hair H o NUTRITION AND TEMPERATURE 28 Ao Preliminary Studies 1* Carbon Sources 2* Hydrogen ion concentration B* Temperature ..... Co Keratin D® Amino Acids E0 Fatty Acids F* Vitamins CONCLUSIONS 0 0 0 0 aocooaoooooooooaoooaoooaaooaooooooooooaooaoo» LITERATURE CITED Oooooooooaooaooaooooooaoaaooooaoooaaoaoooooo V LIST OF TABLES Table Page 1. Growth of P. hortae on hairs from Afro-American and Caucasian females, artificially infested, with observation at various incubation times at 25 C .............. ...•.••»<> 23 2. Dry weight of JP. hortae in basal medium with various carbohydrates added at 5.0 g/1 concentration after 8, 12 and 18 days incubation at 25 C •••••... o . ......*• 29 3. Dry weight of P. hortae in basal medium with various carbohydrates added at 10.0 g/1 concentration after 8, 12 and 18 days incubation at 25 C 30 4* Effect of initial pH on the growth of P, hortae in arginxne-maltose medium after 8 and 12 days incubation at 25 C 33 5. Effect of various temperatures on the growth of P. hortae in malt extract medium after 8, 12 and 18 days incubation at various t e m p e r a t u r e s ••••»•••• 37 6. Effect of various keratin concentrations (percentages) on . the growth of JP. hortae. measured as dry weight (mg), and the final pH at a 25 C incubation temperature........*c 43 7* Growth of P. hortae on basal medium with maltose as the carbon source and five organic nitrogen sources supplied at three concentration levels with 8 days incubation at 25 C .••....«•» 45 8. Growth of P. hortae on basal medium with maltose as the carbon source and five organic nitrogen sources supplied at three concentration levels with 12 days incubation at 25 C .................................. 46 9. Growth of P. hortae on basal medium with maltose as the carbon source and five organic nitrogen sources supplied at three concentration levels with 18 days incubation at 25 C ................ * 10. Effect of various nitrogen sources on the final pH value of the culture medium with maltose as the carbon source and five organic nitrogen sources supplied at three concentration levels with 8 days incubation at 25 C................... 11* Effect of various nitrogen sources on the final pH value of the culture medium with maltose as the carbon source and five organic nitrogen sources supplied at three concentration levels with 12 days incubation at 25 C **•• 49 120 Effect of various nitrogen sources on the final pH value of the culture medium with maltose as the carbon source and five organic nitrogen sources supplied at three concentration levels with 18 days incubation at 25 C *.** 50 13* Growth of P*. hortae after 12 and 18 days incubation at 25 C on basal medium with, arginine supplied at three concentrations with/without the addition of maltose.***** 52 14, Effect of the addition of fatty acids and a vegetable oil to SDA at various concentrations (percentages) on the growth of P, hortae at 25 C «0.* 0 62 15. Growth of P. hortae on basal medium containing three con­ centrations of arginine supplemented with biotin and/or thiamine with 12 days incubation at 25 C ••••••••o******* 66 16. Growth of I\> hortae on basal medium containing three con­ centrations of arginine supplemented with biotin and/or thiamine with 18 days incubation at 25 C *..,.,..,....0** 67 17, Growth of P* hortae on basal medium containing three con­ centrations of arginine supplemented with biotin and/or thiamine with 24 days incubation at 25 C •••.•••,.•••*••* 68 vii LIST OF FIGURES Figures Page 1. Effect of hydrogen ion concentration on the growth of P. hortae in arginine-maltose medium after 8 and 12. days incubation at 25 C ................... 35 2. Effect of various keratin concentrations (percentages) on the growth of P. hortae, measured as linear extension (cm) at various incubation times (days) at 25 C ........ 40 3. Effect of various keratin concentrations (percentages) on the growth of P. hortae, measured as dry weight (mg), with incubation at 25 C ...................... 41 viii LIST OF PLATES : Plate Page .1. Stereoscan electron micrographs of Pe hortae cells cultured in malt extract medium at 25 C and observed at various magnifications.•..••••••• 15 II. Stereoscan electron micrographs of P. hortae cells embedded in an amorphous substance and observed at various magnifications............... 17 III. Photomicrographs of P. hortae cells on human hair segments observed at various magnifications. 19 IV. Stereoscan electron micrographs of artificially infested hairs observed at various magnifications. 22 V. Stereoscan electron micrographs and X-ray point analysis of P. hortae cells on hair obtained from a Bolivian source.......,..,.............©.. 26 VI. Photomicrographs of P. hortae cells cultured in basal medium with arginine and lysine as the nitrogen sources incubated at 25 C 54 VII. Photomicrographs of P. hortae cells cultured in basal medium with cystine and tryptophan as the nitrogen sources incubated at 25 C ..••••.••••••• 56 VIII. Photomicrographs of JP. hortae cells cultured in basal medium with histidine and keratin as the nitrogen sources incubated at 25 C ••••••••••.••• 58 IX. Photomicrographs of JP. hortae cells and crystals produced in basal medium with arginine and cystine as the nitrogen sources incubated at 25 C ....... 60 ix INTRODUCTION Piedraia hortae^- (Brumpt) Fonseca and Ar§a Leao (1928) is the eti­ ological agent of black piedra, a fungal infestation of the hair shaft, characterized by dark, hard, irregularly placed and shaped nodules (ascostromata), hence, the name ''piedra11, which is Spanish for stone. Microscopic examination of the mycelium reveals dark, thick-walled, closely septate hyphae containing numerous chilamydospores or swollen, irregularly shaped cells. According to Castellani and Chalmers (1919) the mycosis has been known in Colombia since remote times, but was not described until 1878 by Dessene. Subsequently, Behrend (1890) reported cases of black piedra in Colombia on hairs of the head being caused by Trichosporum giganteum (Behrend).
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