Whitefly Control and Anti-Microbiological Activities of Essential Oils from Medicinal Plants Found in Fiji Islands
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WHITEFLY CONTROL AND ANTI-MICROBIOLOGICAL ACTIVITIES OF ESSENTIAL OILS FROM MEDICINAL PLANTS FOUND IN FIJI ISLANDS. By Ravneel Rajneel Chand A thesis submitted in fulfillment of the requirements for the degree of Masters of Science Copyright © 2016 by Ravneel Chand School of Biological and Chemical Sciences Faculty of Science, Technology and Environment The University of the South Pacific August, 2016 DECLARATION OF ORIGINALITY I, Ravneel Chand, declare that this thesis is my own work and has not been submitted in any other university. The information provided is best of my knowledge, and information derived from the work of others has been acknowledged in the reference list. Statement by the Principal Supervisor The research work carried by the principal researcher was solely under my supervision and to my best of knowledge. Co-Supervisor Every challenging work needs self-endeavour as well as directions from elders, especially those closest to us. I dedicate this thesis, the fruits of my labour, to my wonderful parents Mr Suresh Chand and Mrs Roshni Chand. ACKNOWLEDGEMENT Prima facea, I am very thankful to the God for the good wellbeing and health throughout my research journey. I acknowledge my sincere gratitude to my Principal supervisor, Associate Professor Anjeela Jokhan and my Co-supervisor Dr. Romila Devi Gopalan for their advice, encouragement and continuous support throughout the study. Working with them was the best part of the research. Thank you once again for the constant motivation and assistance in shaping my transitional skills. A warm thanks to Mr Ashley Dowell and the team from Southern Cross University, Queensland, Australia for assisting me through the identification of compounds in selected essential oils. A special thanks to the Chief Scientist Dr Rajeswara Rao, Dr. Karuna Shanker and the team from Central Institute of Medicinal and Aromatic Plants, India, for sharing their thoughts and ideas throughout the research. I am grateful to the Chief Technician Mr Dinesh Kumar from the Biology Department (USP) for continuous support and assisting me with the plant materials that I possibly have not identified on my own. A special thanks to Dr. Tamara Osborne and Ms Reema Prakash for continuous support through my research journey especially helping out to structure my thesis. Also, I am thankful to Ms Aradhana Deesh from Koronivia Research Station for the assistance in the identification of the selected plant materials and the whitefly species. I am thoroughly grateful to my parents for their continuous support, love and understanding through my research journey, it is the faith that they had on me made me complete this thesis. Finally, I take this opportunity to express my gratitude to one and all, who has directly or indirectly assisted me in completing my Masters study. iv ABSTRACT A variety of plant materials contain essential oils that have extensive bioactivity properties. These properties are attributed to the chemical composition of essential oils. In the current research, chemical composition, whitefly control and anti- microbiological activities of essential oils from five medicinal plants found in Fiji; Cananga odorata (Makasoi), Murraya koenigii (L) Spreng (Curry leaves), Euodia hortensis forma hortensis (Uci), Ocimum tenuiflorum L (Tulsi) and Cymbopogon citratus (Lemon grass) were investigated. Firstly, the selected essential oils were analysed using Gas Chromatography Mass Spectrometry (GC-MS). The identified compounds were classified into groups. For the biological activities, different concentrations of essential oil solutions (0.25%, 0.5% and 5% (v/v)) were subjected to whitefly (Aleurodicus dispersus Russell) control activities in the form of fumigant and repellent test. Essential oils from O. tenuiflorum L were found to be best fumigant agents (100% mortality was achieved at 3 hours after exposure). The significant differences in the mortality for all the tested time (3, 6, 9, 12 and 24 hours) were only shown by O. tenuiflorum L and C. citratus essential oils, as the p<0.05 (5% level of significance). For the repellent test, none of the essential oils obtained 100% repellency based on Repellency index (RI%), however C. citratus and M. koenigii (L) Spreng were found to show the best repelling properties (RI%= 52, 52) compared to the other studied essential oils. In addition, the essential oils exhibited a very interesting antimicrobial profile when tested against five different bacteria and fungi at different concentrations (0.25%, 0.5%, 5%, 25%, 50% and 100% (v/v)). The essential oils from O. tenuiflorum L were considered to have strong antimicrobial properties as it showed the inhibition effect to all test bacteria and fungi. The trends in the chemical constituents of essential oils revealed that the phenolic and alcoholic compounds were major groups of contributors for the tested activities. Thus, these data suggested that essential oils from selected medicinal plants found in Fiji have potential to be employed in pesticide or anti-microbiological activities. v TABLE OF CONTENTS ACKNOWLEDGEMENT ........................................................................................ iv ABSTRACT ................................................................................................................ v TABLE OF CONTENTS .......................................................................................... vi LIST OF TABLES ..................................................................................................... x LIST OF FIGURES ................................................................................................. xii 1.CHAPTER 1: ESSENTIAL OILS ......................................................................... 1 1.0 Introduction ................................................................................................ 1 1.1 Formation of Essential Oils ....................................................................... 1 1.2 Components of Essential Oils .................................................................... 3 1.2.1 Terpenes hydrocarbons .......................................................................... 3 1.2.2 Oxygenated Compounds ........................................................................ 7 1.2.3 Ethers...................................................................................................... 8 1.2.4 Aldehydes ............................................................................................... 9 1.2.5 Ketones ................................................................................................. 10 1.2.6 Organic acids and esters ....................................................................... 10 1.2.7 Oxides .................................................................................................. 11 1.3 Extraction of Essential Oils ..................................................................... 11 1.3.1 Distillation ............................................................................................ 11 1.3.2 Solvent extraction................................................................................. 13 1.3.3 Enfleurage ............................................................................................ 13 1.4 Methods for Analysis of Chemical Constituents ..................................... 13 1.4.1 Gas-Liquid Chromatography ............................................................... 14 1.4.2 Gas Chromatography-Mass Spectrometry ........................................... 14 1.5 Common Uses of Essential Oils .............................................................. 14 1.5.1 Essential Oils Used by Plants ............................................................... 14 vi 1.5.2 Essential Oils Used by Humans ........................................................... 15 1.6 Purpose of this study ................................................................................ 18 2.CHAPTER 2: CHEMICAL ANALYSIS OF ESSENTIAL OILS FROM SELECTED MEDICINAL PLANTS FOUND IN FIJI. ....................................... 20 2.0 Introduction .............................................................................................. 20 2.1 Background .............................................................................................. 21 2.1.1 Description and Common Uses of Selected Medicinal Plants Found in Fiji. .............................................................................................................. 21 2.2 Methodology ............................................................................................ 26 2.2.1 Collection of Plant Materials ............................................................... 26 2.2.2 Extraction of Essential Oils .................................................................. 26 2.2.3 Analysis of Chemical Constituents ...................................................... 27 2.3 Results ...................................................................................................... 28 2.3.1 Physical Properties ............................................................................... 28 2.3.2 Gas Chromatography-Mass Spectrometry (GC-MS) Analysis ............ 29 2.4 Discussion ................................................................................................ 40 2.4.1 Cananga odorata (Makosoi) ................................................................. 40 2.4.2 Murraya koenigii (Curry leaves) .........................................................