Effect of Rutaceae plant's essential oils and leaf extracts on dermatophytic fungal cell morphology

174 Pages·2016·2.63 MB·English

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Effect of Rutaceae plant’s essential oils and leaf extracts on
dermatophytic fungal cell morphology: a hope for the development of
an effective antifungal from natural origin

Submitted in fulfillment of the academic
requirements for the degree of
DOCTOR OF PHILOSOPHY

By
OLUFUNKE OMOWUMI FAJINMI

Research Centre for Plant Growth and Development
School of Life Sciences
College of Agriculture, Engineering and Science
University of KwaZulu-Natal, Pietermaritzburg

May 2016
Pictures sourced from google
A healthy, glowing, beautiful skin….the pride of
every woman

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Table of Contents
STUDENT DECLARATION ................................................................................................... v
DECLARATION BY SUPERVISORS ....................................................................................... vi
COLLEGE OF AGRICULTURE ENGINEERING & SCIENCE DECLARATION 1- PLAGIARISM ........ vii
ACKNOWLEDGEMENTS .................................................................................................. viii
COLLEGE OF AGRICULTURE ENGINEERING & SCIENCE DECLARATION 2- PUBLICATIONS ....... x
LIST OF FIGURES .............................................................................................................. xi
LIST OF TABLES ............................................................................................................... xii
LIST OF ABBREVIATIONS ................................................................................................ xiv
ABSTRACT ...................................................................................................................... xv
CHAPTER 1. INTRODUCTION.............................................................................................. 1
1.1. South Africa and its floral biodiversity ............................................................................. 1
1.2. Aesthetic value and healing power of the “Cape green gold” ......................................... 1
1.3. Aromatic plants meeting the needs of humans from time immemorial ......................... 2
1.4. Use of aromatic plants in aromatherapy and human health since “the days of old” ...... 3
1.5. The Rutaceae, a family of green aromatics in the treasure-box of the Rainbow Nation
4
1.6. Significance of the study ................................................................................................... 7
1.7. Aims and objectives .......................................................................................................... 9
CHAPTER 2. LITERATURE REVIEW .................................................................................... 10
2.1. HIV/AIDS: a trauma to the human race .......................................................................... 10
2.2. The human skin, a vital irreplaceable organ................................................................... 10
2.3. Skin diseases: a health burden across the globe ............................................................ 12
2.4. Skin diseases in Africa, the culprits and way forward .................................................... 13
2.4.1. The increased burden of skin diseases resulting from HIV/AIDS ........................... 14
2.4.2. Prevalence of skin diseases in HIV/AIDS individuals and the microorganisms
responsible for the infections/diseases ........................................................................... 15
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2.4.3. Fungi as major causative agents of skin diseases in humans ................................. 16
2.4.4. Classification of Human mycoses ........................................................................... 19
2.4.5. Nail infections caused by fungi ............................................................................... 23
2.4.6. Treatment of Mycosis ............................................................................................. 24
2.5. Plants, a source of therapeutic compounds ................................................................... 33
2.5.1. Plants as a potential source of antifungals ............................................................. 34
2.5.2. Isolation of coumarins and other bioactive compounds from Rutaceae species .. 35
2.5.3. Rutaceae species used in traditional medicine for skin related diseases and
infections .......................................................................................................................... 39
2.5.4. Sources and components of essential oil ............................................................... 41
2.6. Pathogenesis of skin diseases ......................................................................................... 45
CHAPTER 3. ANTIOXIDANT ACTIVITY OF SEVEN RUTACEAE PLANT LEAF EXTRACTS ........... 51
3.1. Introduction .................................................................................................................... 51
3.2. Materials and methods .................................................................................................. 53
3.3. Results ............................................................................................................................ 54
3.4. Discussion ....................................................................................................................... 56
3.5. Conclusions ..................................................................................................................... 58
CHAPTER 4. ANTIFUNGAL ACTIVITY OF SEVEN RUTACEAE PLANT LEAF EXTRACTS AGAINST
TRICHOPHYTON RUBRUM, T. MENTAGROPHYTES AND MICROSPORUM GYPSEUM .......... 59
4.1. Introduction .................................................................................................................... 59
4.2. Materials and methods .................................................................................................. 60
4.3. Results ............................................................................................................................ 61
4.4. Discussion ....................................................................................................................... 65
4.5. Conclusions ..................................................................................................................... 67
CHAPTER 5. ANTIFUNGAL ACTIVITY OF ESSENTIAL OILS DERIVED FROM RUTACEAE PLANT
LEAVES AGAINST THREE SKIN DISEASE FUNGAL STRAINS ................................................. 68
5.1. Introduction .................................................................................................................... 68
5.2. Materials and methods .................................................................................................. 69
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5.3. Results ............................................................................................................................ 70
5.4. Discussion ....................................................................................................................... 74
5.5. Conclusions ..................................................................................................................... 76
CHAPTER 6. ANALYSIS OF THE ESSENTIAL OIL COMPONENTS OF SEVEN RUTACEAE SPECIES
...................................................................................................................................... 77
6.1. Introduction .................................................................................................................... 77
6.2. Materials and methods .................................................................................................. 78
6.3. Results ............................................................................................................................ 78
6.4. Discussion ....................................................................................................................... 84
6.5. Conclusions ..................................................................................................................... 86
CHAPTER 7. ANTIFUNGAL ACTIVITY OF ESSENTIAL OILS OF AGATHOSMA BETULINA AND
COLEONEMA ALBUM AND ITS EFFECT ON THE MORPHOLOGY OF T. RUBRUM AND T.
MENTAGROPHYTES ........................................................................................................ 87
7.1. Introduction .................................................................................................................... 87
7.2. Materials and methods .................................................................................................. 88
7.2.1. Antifungal activity of essential oil volatiles and essential oil analysis .................... 88
7.2.2. Effect of essential oil volatiles on morphology of fungal mycelia .......................... 89
7.3. Results ............................................................................................................................ 89
7.3.1. Antifungal activity of essential oil volatiles and essential oil analysis .................... 89
7.3.2. Effect of essential oil volatiles on morphology of fungi mycelia ............................ 90
7.4. Discussion ....................................................................................................................... 97
7.5. Conclusions ................................................................................................................... 102
CHAPTER 8. GENERAL DISCUSSION ................................................................................ 104
CHAPTER 9. CONCLUSIONS AND RECOMMENDATIONS .................................................. 109
CHAPTER 10. REFERENCES ............................................................................................. 111

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STUDENT DECLARATION

I, Olufunke Omowumi Fajinmi, 210556317 (student number) declare that:
i. The research reported in this dissertation, except where otherwise indicated,
is the result of my own endeavours in the Research Centre for Plant Growth
and Development, School of Life Sciences, College of Agriculture,
Engineering and Science, University of KwaZulu-Natal, Pietermaritzburg.
ii. This dissertation has not been submitted for any degrees or examination at
any other university.
iii. This thesis does not contain data, figures or writing, unless specifically
acknowledged, copied from other researchers.
iv. Where I have reproduced a publication of which I am an author or co-author,
I have indicated which part of the publication was contributed by me.

Signed at on the day of 2016.
__________________________
SIGNATURE

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DECLARATION BY SUPERVISORS

We hereby declare that we acted as Supervisors for this PhD student:
Student’s Full Name: Olufunke Omowumi Fajinmi
Student Number: 210556317
Thesis Title: Effect of Rutaceae plant’s essential oils and leaf extracts on
dermatophytic fungal cell morphology: a hope for the development of an effective
antifungal from natural origin.
Regular consultation took place between the student and us throughout the
investigation. We advised the student to the best of our ability and approved the
final document for submission to the College of Agriculture, Engineering and
Science, Higher Degrees Office for Examination by the University appointed
Examiners.

______________________________
SUPERVISOR: PROFESSOR. J. VAN STADEN

______________________________
CO-SUPERVISORS: PROFESSOR. J. F. FINNIE

______________________________
DR M.G. KULKARNI

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COLLEGE OF AGRICULTURE ENGINEERING & SCIENCE
DECLARATION 1- PLAGIARISM

I, Olufunke Omowumi Fajinmi declare that:

1. The research reported in this thesis, except where otherwise indicated, is my
original research.
2. This thesis has not been submitted for any degree or examination at any
other university.
3. This thesis does not contain any other persons’ data, pictures, graphs or
other information, unless specifically acknowledged as being sourced from
other persons.
4. This thesis does not contain any other persons’ writing, unless specifically
acknowledged as being sourced from other researchers. Where other written
sources have been quoted , then:
a. Their words have been re-written but the general information
attributed to them has been referenced.
b. Where their exact words have been used, then their writing has been
placed in italics and inside quotation marks, and referenced.
5. This thesis does not contain text, graphics or tables copied and pasted from
the internet, unless specifically acknowledged, and the source being detailed
in the thesis and in the References sections.
6. The essential oil analysis and antioxidant activity recorded in this study was
carried out by our collaborators in Czech Republic.

Signed _____________________

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ACKNOWLEDGEMENTS

I give thanks to the Almighty God for giving me the opportunity to complete
this study with ease. When my strength and wisdom failed me, his divine presence
kept me going. He started the good work and completed it in due season. I would
like to express my sincere gratitude to my supervisor Prof Van Staden, for his
words of encouragement and inspiration during the course of this study, you are
indeed a great mentor. I appreciate my co-supervisors; Prof Finnie for his constant
support, kind efforts and words of encouragement; Dr Kulkarni for his words of
encouragement.
Many thanks to all members of the Research Centre for Plant Growth and
Development for the friendship we have shared over the years. Special thanks to Dr
Becki Ncube and ‘almost’ Dr Devashan Naidoo. Mrs Warren is highly appreciated
for her help as regards administrative matters. My sincere appreciation goes to Mrs
Alison Young for her assistance as regards seeds and plant collection. I appreciate
our Czech collaborators; Prof Dolezal and his wife (Ivana), Petre Tarkowski and
others. Thanks to Tibor Beres for his help while I was in Czech. The staffs of
Electron Microscopy Unit of UKZN (Subashen, Lorika Beukes and Cynthia) are
highly appreciated for their help with electron microscopy work.
I appreciate Pastor and Mrs Olorunda of Dunamis faith assembly for their
prayers, words of encouragement, love and support. Many thanks to my guardian
Mr and Mrs Fatokun for their support since my MSc days. I appreciate Dr Siyanbola
for his fatherly advice and encouragements. To Dr and Dr Mrs Ogunrombi thanks
for your love. To my wonderful friend Abimbola Ajetumobi thanks for asking how far
and giving me a listening ear. To my friend Renate Makekita Ntumpi, thanks for
escorting me to school during late hours, you are indeed a blessing.
Special thanks to my wonderful parents and brothers for their love, financial
support, encouragement and understanding, you are my greatest assets. If I have
to come back to this world the second time, I would choose Mr and Mrs Fajinmi as
my parents as their love and dedication is second to none. I remember all the
sacrifices you made for me and my siblings to have good education. You sacrificed
your time, money and efforts to make us who we are today. You are angels in
human clothing!
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To my darling husband Olaoluwa Olarewaju who I met during the course of
this degree, you are the best thing that ever happened to me. The academic
challenges I encountered during the first semester of my PhD brought us together
as friends and we graduated to best friends and before I knew it, we got married!
Your love and support kept me going during the tough moments as a student.
Thanks for being there always, you are totally amazing. I cannot find the right words
to accurately express how much you mean to me!
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