THE CONTROL OF RED SPIDER MITES ON TOMATOES USING NEEM AND SYRINGA EXTRACTS by NEPHIOUS JAMES KAMALENJE MWANDILA Submitted in accordance with the requirements for the degree of DOCTOR OF PHILOSOPHY in the subject ENVIRONMENTAL MANAGEMENT at the UNIVERSITY OF SOUTH AFRICA SUPERVISOR: PROF J OLIVIER Co - SUPERVISORS: DR D VISSER PROF D.C MUNTHALI NOVEMBER 2009 i Student No: 3660-961-7 Declaration I declare that THE CONTROL OF RED SPIDER MITES ON TOMATOES USING NEEM AND SYRINGA EXTRACTS is my own work and that all the sources that I have used or quoted have been indicated and acknowledged by means of complete references -SIGNATURE (MR N.J.K MWANDILA) DATE ii Abstract The efficacy of Neem (Azadirachta indica A Juss) and Syringa (Melia azedarach L.) against red spider mites (RSM) life phases (adult, nymphs and eggs) was assessed at different concentrations (0.1%, 1%, 10%, 20%, 50%, 75%, 100%) and at exposure time of 24, 48 and 72 hours using tomato leaf dip assays on water agar in plastic Petri dishes Tomato plants were grown in the greenhouse as a source of leaves and for the greenhouse trial A Greenhouse trial was carried out to simulate field conditions Neem seeds were sourced from Botswana, India, and Zambia Syringa seeds were sourced from Botswana and South Africa Laboratory and Greenhouse trials were carried out at the Agriculture Research Council, in the Vegetable and Ornamental Institute laboratories and green houses in Pretoria South Africa Data was analysed by using the GenStat statistical program Overall results of both Neem and Syringa assays indicated that all levels of concentrations and time of exposure had significant effects on mortalities of adult RSM and compared significantly with commercial acaricides (Abamectin-plus, Hunter and Selecron) Both Neem and Syringa caused significant mortalities at low concentration of 0.1% as early as 24 hours of exposure Both Neem and Syringa assays had significant mortalities on RSM nymphs as early as 24 hours and with longer periods of exposure Both Neem and Syringa had significant effects on the hatching of RSM eggs at 48 hours and 72 hours of exposure In general, effects occurred in a dose (concentration) dependent manner Based on the findings and evidence in the literature, Neem and Syringa extracts could be useful as botanical acaricides in the control of red spider mites (RSM) on tomatoes iii Key terms: Neem (Azadirachta indica); Syringa (Melia azedarach); Azadirachtin compound; Azadirachtin standard; High Performance Liquid Chromatography (HPLC); Samples; homogenous; Analysis of Variance; Randomized Designs; Abamectin-plus; Hunter; Selecron; liquid–cooling agar; red spider mites; life cycle; heterogeneous; antifungal; antibacterial; antifeedant; larva; protonymph; deutonymph.; Globalisation; food production; food consumption; cereals; vegetables; pests; pesticides; limonoids iv Dedication This thesis is dedicated to: • the memory of my mother Elinala and my father James Kamalenje Mwandila-two individuals who made a lasting impact on my life by bringing me up and affording me an opportunity to attend school • my late son Daniel who kept encouraging me till he met his death by a robber’s bullet • my eldest son Harvesto Malombo and his siblings- Elizabeth, Tiyezye, Bangala, Newton, Esnart, Kasimba and Khumbo v Acknowledgements I am grateful to members of my family for their unstinting support throughout the duration of this study Special thanks are due to my children Harvest-Malombo, Daniel (late), Elizabeth, Tiyezye, Esnart, Bangala, Newton, Kasimba, Khumbo and my nephew Conerlius who always kept me smiling and feeling lighthearted in those anxious moments of thesis-writing when I was very sick I also wish to thank, most profoundly, my promoter, Professor Jana Olivier, for her encouragement, her intelligence, her care, and above all, her nurturing approach to the whole process of supervision For me, working with her was always a source of much immense joy even when I was seriously sick and quitting seemed to be the only option especially after losing my son Daniel I want to thank • Dr Diedrich Visser my co-promoter, for his support and advice throughout the time when I carried out the Laboratory bioassays at the Vegetable and Ornamental Plant Institute (VOPI) and his valuable comments during the process of supervision • Professor David Munthali my co-promoter, for his advice while in Botswana • The staff at the Agricultural Research Council’s (ARC) Vegetable and Ornamental Plant Institute, Roodeplaat in particular Dr Nolwazi Mkize and Sakki Sambo for their support and help in purchasing several useful materials that were used in the bioassays during my research experiments • Nanga Irrigation Research Station - Zambia for harvesting and providing me with Neem seeds vi • Mrs Jeyasseeli Michael for sourcing me Neem seeds from India • My heart goes to all the VOPI staff for welcoming me as part of their team • Dr Gerhard Prinsloo for providing me with much advice on research activities, especially in the handling of High Performance Liquid Chromatography (HPLC) • Subject Librarian, Mrs Leanne Tracy Brown for her efficiency, integrity and charm • Marie Smith, Elise Robbertse and Poloko Chepete for helping with statistics • Charnie Creamer ARC - Plant Protection Research Institute, for the identification of the mites used in the bioassays • Dr Fetson Kalua of UNISA for making my life easier while in South Africa by providing accommodation and valuable advice and encouragement • G.N Mthombeni for financial assistance during the time I desperately needed to pay my fees at UNISA • Letsholo Bongalo for helping me with statistical graphs during my write up • Not forgetting other friends and individuals too many to mention who stood by me when getting the thesis done Finally, I wish to acknowledge the Department of Agriculture and Environmental Sciences and the Agricultural Research Council (ARC) for funding part of this project Yebo/ Thank you vii TABLE OF CONTENTS Declaration ii Abstract iii Dedication iv Acknowledgements v Table of contents vii Abbreviations viii Appendices iv Chapter General Introduction 1.1 Background to the study 1.1.1 Overview on food production 1.1.1.1 Food production and consumption patterns 1.1.1.2 World cereal production 1.1.1.3 World vegetable production 1.1.1.4 Vegetable production in the SADC region 11 1.1.1.5 Vegetable production in Botswana 12 1.1.1.5.1 Tomato production in Botswana 13 1.1.2 14 Factors affecting food production 1.1.2.1 Crop pests 14 1.1.2.2 Types of pests 15 1.1.2.3 Pesticides and their problems 16 1.1.2.4 Possible solutions: Botanical pesticides 18 viii 1.2 Summary and problem statement 20 1.3 Aim 22 1.3.2 Objectives of the study 22 1.4 Chapter layout 22 Chapter Literature Review 23 2.1 Introduction 23 2.2 Red spider mites (RSM) 24 2.2.1 Taxonomy 24 2.2.2 Life cycle 25 2.2.3 Damage caused by red spider mites 30 2.3 Control measures 31 2.3.1 Natural control measures 31 2.3.2 Chemical control measures 32 2.3.3 Biological control measures 34 2.4 Botanicals as an option 35 2.4.1 Introduction 35 2.4.1 Neem (Azadirachta indica A Juss) as a botanical pesticide 36 2.4.2.1 Origin and distribution of Neem (Azadirachta indica A Juss) 38 2.4.2.2 Uses as pesticides 38 2.4.2.3 Chemical composition 39 2.4.3 41 Syringa (Melia azedarach L) as a botanical pesticide 2.4.3.1 Origin and distribution of Syringa (Melia azedarach L.) 44 ix 2.4.3.2 Uses as pesticides 44 2.4.3.3 Chemical composition 45 2.2.3.4 Summary 46 Chapter Research Design and Methodology 47 3.1 Introduction 47 3.2 Study area 48 3.3 Materials and Methods 49 3.3.1 Collection of seeds 49 3.3.1.1 Neem seed collection 49 3.3.1.2 Syringa seed collection 49 3.3 Determination of azadrachtin content of seeds 50 3.3.2.1 Preparation of standards 50 3.3.2.2 Extraction of the active constituents from Neem and Syringa seeds 50 3.3.3 Experimental plants 51 3.3.4 Data collection 52 3.3.4.1 Laboratory bioassays 52 3.3.4.2 Testing the effect of Neem and Syringa extracts on adult RSM 52 3.3.4.3 Testing the effect of Neem and Syringa extracts on nymph red spider mites 54 3.3.4.4 Testing the effect of neem and syringa extracts on red spider mite eggs 55 3.3.5 56 Greenhouse trial x Blumel, S & Hausdorf, H (2002) Results of the 8th and 9th IOBC joint pesticides testing programme: In: H Vogt, & U Heimbach (Eds.), Proceedings of the Meeting at San Michele All’Adige, Persistence Test with Phytoseiulus persimilis vol 25 Athias Henriot (Acari: Phytoseiidae) Bulletin- OILB-SROP (pp 43-51) Trento, (Italy) Bohmont, B L (1997) The Standard Pesticide: Users Guide Prentice Hall Upper Saddle River, New Jersey Bok, I., Madisa M., Machacha, D, Moamongwe M & More, K (2006) Manual for Vegetable Production in Botswana Ministry of Agriculture, Botswana Boeke, S J., Van Loon J J A, Van Huis A., Kossou & Dicke, M (2001) The use of plant material to protect stored leguminous seeds against seed beetles: a review, Wageningen University papers 2001-3, The Netherlands Bolland, H R & Valla, F (2000) First record of the spider mite Tetranychus evansi (Acari: Tetranychidae) from Portugal Entomologische Berichten 60 (9), 180 Bolland, H R., Guitierrez, J & Flechtmann, C H W (1998) World Catalogue of Spider Mites Family (Acari: Tetranychidae) (pp 392) Brill, Leiden, Boston, Köln Bonnano, A (Ed.) 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INTRODUCTION 1.1 Background to the study 1.1.1 Introduction This chapter presents the background of the study which includes an overview of global food production and consumption, the problem statement, the aim and objectives of the study and the significance of study 1.1.1.1 Overview on food production Low agriculture productivity and declining production efficiencies pose a threat to global food production... bio-fuels rather than for food (WRR, 2007) In an article entitled The world’s choice: food or bio-fuels” which appeared in The Sunday Times of March 9 2008 page 6, Beddington (2008) pointed out that the world today is concentrating on the production of crops for bio-fuels rather than for food supply, thereby contributing to the decline in cereal production In the 2006 annual assessment of the global... fruits There are a number of other pests that cause damage to tomatoes and reduce yields These include pests like tomato semi-looper (Chrysodeixis acuta), and nematodes (Meloidogyne species) (Kasozi et al., 1999) However, one of the most common pests of tomato is the red spider mite (Wikipedia, 2007) Bok et al., (2006) reported that various species of red spider mites attack the tomato crop in Botswana reducing... Dead adult mites on Syringa leaf extracts 1 D Dead mite nymph on Neem seed extracts 1 E Dead mite nymph on Syringa seed extracts 1 F Number of hatched eggs on Neem seed extracts 1 G Number of hatched eggs on Syringa seed extracts 1 H The greenhouse trial APPENDIX 2 Calculation of percentage efficacy of Neem and Syringa seed extracts APPENDIX 3 A protocol for the control of red spider mites using Neem... who cannot afford the expensive and intricate usage of conventional pesticides and most of the food that they produce is consumed locally One of the main vegetable food crops produced in Botswana and in the world is tomato However, this crop is heavily attacked by red spider mites (Bok et al., 2006) It is important therefore that a solution be sought A solution could be the development of a botanical... within the mentioned countries has changed over the past few years, with greater imports of fruits and vegetables coming from SADC countries (Table 1.4) and other developing countries in the southern hemisphere 10 1.1.1.4 Vegetable production in the SADC region Until recently, vegetable production was an ignored and little-known industry in the SADC region (Mnzava, 1990) For a long time, the production of. .. (2005), 125 million tonnes of tomatoes were produced in the world in 2005 The largest producers of tomatoes (in tonnes) were: China, accounting for about one-fourth of the global output; the United States is second, with Turkey third In South Africa, tomatoes are among the most important and highly valued horticultural products (Louw, 2005) Louw (2005), has observed that in 2004 tomato production in South