University of Wollongong Research Online University of Wollongong Thesis Collection 1954-2016 University of Wollongong Thesis Collections 1985 The numerical taxonomy of pathogenic species of candida William James Crozier University of Wollongong Follow this and additional works at: https://ro.uow.edu.au/theses University of Wollongong Copyright Warning You may print or download ONE copy of this document for the purpose of your own research or study. The University does not authorise you to copy, communicate or otherwise make available electronically to any other person any copyright material contained on this site. You are reminded of the following: This work is copyright. Apart from any use permitted under the Copyright Act 1968, no part of this work may be reproduced by any process, nor may any other exclusive right be exercised, without the permission of the author. Copyright owners are entitled to take legal action against persons who infringe their copyright. A reproduction of material that is protected by copyright may be a copyright infringement. A court may impose penalties and award damages in relation to offences and infringements relating to copyright material. Higher penalties may apply, and higher damages may be awarded, for offences and infringements involving the conversion of material into digital or electronic form. Unless otherwise indicated, the views expressed in this thesis are those of the author and do not necessarily represent the views of the University of Wollongong. Recommended Citation Crozier, William James, The numerical taxonomy of pathogenic species of candida, Master of Science thesis, Department of Biology, University of Wollongong, 1985. https://ro.uow.edu.au/theses/2618 Research Online is the open access institutional repository for the University of Wollongong. For further information contact the UOW Library: [email protected] THE NUMERICAL TAXONOMY OP PATHOGENIC SPECIES OP CANDIDA. A thesis submitted in partial fulfilment of the requirements of the degree of MASTER OF SCIENCE from THE UNIVERSITY OF WOLLONGONG by WILLIAM JAMES CROZIER B.Appl.Sc. (Hons) UNIVERSITY OF WOLLONGONG ARY BIOLOGY DEPARTMENT JUNE, 1985 898725 DECLARATION The work presented in this thesis, in fulfilment of conditions governing the award of the degree of Master of Science, was carried out by me between September 1978 and February 1982 externally at the Mycology Laboratory of the Wollongong Hospital through the Biology Department, University of Wollongong, Wollongong, Australia and has not been submitted to any other university or institution for a higher degree* William James Crozier, J*P June, 1985 - i - SUMMARY. One hundred and eighty-one isolates of Candida comprising C. albicans ( 58.6 % ), C. parapsilosis ( 25.2 % ), C. tropicalis ( 10.5 % )* C. krusei ( 4*4 % )* C. guilliermondii ( 2.2 % ), C. pseudotropicalis ( 0.6 % ) and C. ravauttii ( 0.6 f» ), were examined for 26 physiological characteristics. The characters obtained were subjected to 5 different clustering analyses ( MULTBET, MULCLAS using " incremental sum of squares " fusion strategy and MULCLAS using " group average " fusion strategy ). There appears to be little difference between the groupings obtained using a set of Candida species freshly isolated from clinical specimens and those obtained for the same yeast species maintained for many years on artificial media and under different conditions in a culture collection. Fewer characters were used in this study than are conventionally used in taxonomic studies of yeasts. The reasons were that it was intended to use rapid methods which were currently carried out in routine clinical microbiology laborat­ ories, or which could easily and cheaply adapt to the routine laboratory situation. Although the set of characters used in this study were fewer in number than those used in other taxonomic studies, the groupings obtained were comparable. In fact, this study suggests that, as a limited number of Candida species is encountered in the clinical situation, far fewer characters than were tested here may be used for satisfactory grouping. - ii - Finally, evidence was obtained to suggest that there was no statistically significant difference between the various species of Candida tested in the presence of Amphotericin B and 5 - Fluorocytosine. This study suggests that differences in antifungal tolerance by various species of pathogenic Candida may not constitute characters which are suitable for taxonomic applications. - iii - ACKNOWLEDGEMENTS. I would like to express my appreciation to my supervisor» Professor A#D# Brown» for his encouragement» guidance and helpful discussion throughout this project# Initially» Dr# Robert Cortis-Jones (as Director of Pathology) and then, Dr# Prank Jennis (as Director of Microbiology), at The Wollongong Hospital, kindly allowed me the time and facilities to carry out this study within the Mycology Laboratory. The assistance of the following people is also gratefully acknowledged : Dr# W.T. Williams, Australian Institute of Marine Science, for running the clustering analyses; Dr# V. Drastik, Mathematics Department, University of Wollongong, for running the analyses of variance; Dr. Peter Moran, initially Biology Department, University of Wollongong and then, Australian Institute of Marine Science, for helpful advice on numerical taxonomy methods; Dr# Prank Jennis, Director of Microbiology, The Wollongong Hospital, for running the correlation coefficient programmes and for helpful discussion; Dr. Rob Whelan, Biology Department, and Dr. John Reichelt, Roche Research Institute of Marine Pharma­ cology, for advice and discussion; Miss E. Meyer, Biology Dep­ artment, University of Wollongong, for the photographs used in figures 23* 24 and 25. Finally, I wish to sincerely thank my wife, Sylvia, without whose encouragement and understanding this study would not have been completed. iv - TABLE OF CONTENTS. Page. SUMMARY i ACKNOWLEDGEMENTS iii TABLE OP CONTENTS iv LIST OP TABLES vii LIST OP FIGURES viii INTRODUCTION 1. 1.1 The Classification of Fungi 1. 1.2 Brief History of Classification of Fungi 1. 1.3 The Classification of Yeasts 3» 1.4 Criteria used in Classification of Yeasts 8. 1.5 Criteria used in Classification of Genus : Candida 10. 1.6 History of Medical Mycology 12. 1.7 History of Yeasts within Medical Mycology 13* 1.8 History of Genus : Candida within Medical Mycology 20. 1.9 Basis for Ascribing * Pathogenicity 1 to 22. Certain Candida Species 1.10 * Candida ' versus 1 Torulopsis * 26. 1.11 Numerical Taxonomy of Yeasts 27. 1.12 Numerical Taxonomy Studies of Pathogenic 28. Candida Species MATERIALS AND METHODS 30. 2.1 Yeasts 30. 2.2 Characters used for Numerical Taxonomy 30. 2.3 Additional Tests 31« - v - TABLE OF CONTENTS ( continued )... Page 2.4 Media used for testing Characters 55. 2.5 Effect of Different Antifungal Substances on 55. Pathogenic Candida Species 2.6 Medium used for testing Antifungal Effects 56. 2.7 Cultures 57. 2.8 Yeast Growth in the Presence of Antifungals 58. 2.9 Purity 58. 2.10 Rate of Growth of Yeast Strains 59. 2 .11 Analysis of Exponential Growth Rates for each 59. Yeast to check for Possible Potential Species Characteristics 2.12 Taxonomy / Numerical Analysis 40. 2.15 Fusion Strategies 41. 2.14 Similarity Coefficients 42. RESULTS 44. 5.1 Numerical Taxonomy 44. 5.2 Effect of Amphotericin B and 5 - Fluorocytosine 57. (5 - FC) on Growth Rates of C. albicans, C. tropicalis and C. parapsilosis (i) Plots of log Absorbance (A) v s . Time 57. (ii) Growth Rates 60. (iii) Correlation Coefficients 61. (iv) Statistical Analysis 62. DISCUSSION OP RESULTS 64. 4.1 Numerical Taxonomy 64. 4.2 Substrate Choice as a Basis for Differentiation 68. of Pathogenic Candida Species - vi - TABLE OF CONTENTS ( continued )... Page. 4»3 Growth Plots 7 1 . CONCLUSIONS 76. APPENDIX 1 : GLOSSARY 78. APPENDIX 2 s Binary Data Accumulated from the Set 80. of 181 Strains of Yeasts Tested for 26 Characters APPENDIX 3 : Figures 1, 2, 3» 4* 5$ 6, 7» 8» 9$ 10, 94. 1 1 , 12 , 13, 22, 23, 24, 25, 26, 27 BIBLIOGRAPHY 112. ADDENDUM 130. - vii - LIST OF TABLES. Page# TABLE 1# Earliest known reports of the principal 15* types of Candidosis TABLE 2. Industrially-important Saccharomyces 19. species - Earliest accredited description * TABLE 3* First Taxonomic Description of "Pathogenic” 21. Candida species TABLE 4. Binary Data Accumulated from the Set of 80. 181 Strains of Yeasts tested for 26 Characters TABLE 5* Composition of Groups Generated by 49* Classification A TABLE 6. Composition of Groups Generated by 52. Classification B TABLE 7* Composition of Groups Generated by 55* Classification C TABLE 8. Exponential Growth Rate of Yeast Strains 60. in the Presence of Amphotericin B TABLE 9* Exponential Growth Rate of Yeast Strains 60. in the Presence of 5-Fluorocytosine TABLE 10. Correlation Coefficients for Plots of log ^ Absorbance (A) vs. Time; (Amphotericin B) TABLE 11. Correlation Coefficients for Plots of log 62. Absorbance (A) vs. Time; (5-Fluorocytosine) - viii LIST OF FIGURES. Page FIGURE 1. Candida tropicalis 94. FIGURE 2. Candida parapsilosis 95. FIGURE 3. Candida guilliermondii 96. FIGURE 4. Candida albicans 97. FIGURE 5. Torulopsis glabrata ( Candida glabrata) 98. FIGURE 6. Torulopsis famata ( Candida famata ) 99. FIGURE 7. Candidiasis; kidney abscess; caused by 100. C. albicans FIGURE 8. Candidiasis; peptic ulcer; caused by 101. C. albicans FIGURE 9. Onychomycosis; thumb nail; caused by 102. C. albicans FIGURE 10. Onychomycosis; finger nail; caused by 103. T. Candida ( C. famata) FIGURE 11. Onychomycosis; toe nail; caused by 104. C. ravauttii FIGURE 12. Vaginal Candidiasis; caused by 105. C. albicans FIGURE 13. Germ-Tubes of C. albicans 106. FIGURE 14. Dendogram of Classification A. (i) 45. FIGURE 15. Dendogram of Classification B. (i) 46. FIGURE 16. Dendogram of Classification C. (i) 47. FIGURE 17. Dendogram of Classification A. (ii) 50. FIGURE 18. Dendogram of Classification B. (ii) 53. FIGURE 19. Dendogram of Classification C. (ii) 56. - ix - LIST OF FIGURES ( continued)• • • Page. FIGURE 20 Plot of log Absorbance (A) vs. Time 58. C. albicans grown in varying Concentrations of 5 - FC FIGURE 21 Plot of log Absorbance (A) vs.