Table of Contents TABLE OF CONTENTS Table of contents...................................................................................................................... 1 Acknowledgments.................................................................................................................... 3 Riassunto ................................................................................................................................. 4 Abstract .................................................................................................................................... 8 1. GENERAL INTRODUCTION ............................................................................................. 12 1.1 Adephaga..................................................................................................................... 12 1.2 Carabid Beetles ........................................................................................................... 12 1.2.a Breeding and survival ........................................................................................... 14 1.2.b Carabid “life form” and diet ................................................................................... 15 1.2.c Carabid digestion .................................................................................................. 18 1.2.d Carabid Foraging Behaviour................................................................................. 19 1.2.e Carabid as pest antagonist ................................................................................... 19 1.3 Olive groves ................................................................................................................. 20 1.3.a Olive Pests............................................................................................................ 22 1.3.b The Olive fly Bactrocera (Dacus ) oleae (Rossi 1790) .......................................... 22 1.3.c Carabids/olive fly interaction ................................................................................. 25 RESEARCH GOAL ................................................................................................................ 28 2. INTRODUCTION TO THE PHASES OF THE STUDY...................................................... 29 2.1 Preliminary test ............................................................................................................ 29 2.2 Quantitative test ........................................................................................................... 29 2.3 Microscopical gut content analysis .............................................................................. 30 2.4 Molecular gut content analysis..................................................................................... 30 2.5 Carabid daily rhythm .................................................................................................... 31 2.6 Carabid density in olive grove...................................................................................... 32 2.7 Simulation of carabid predation on B. oleae ................................................................ 32 3. MATERIALS AND METHODS........................................................................................... 33 3.1 Sampled olive grove .................................................................................................... 33 3.2 Laboratory rearing conditions ...................................................................................... 34 Grafic 3.1- Climogram recorded by the thermopluviometric station of Mirto during the period 1999-2005 ........................................................................................................................................ 34 3.2.a Predator carabid beetles studied .......................................................................... 34 3.3 Prey used and food preparation................................................................................... 38 _____________________________________________________________________________________ Rosalba Odoguardi PhD thesis 1 Table of Contents 3.4 Preliminary tests .......................................................................................................... 38 3.5 Quantitative tests ......................................................................................................... 39 3.5.b Experimental conditions........................................................................................ 39 3.5.c Behavioural analysis ............................................................................................. 40 3.5.d Food preferences.................................................................................................. 40 3.6 Microscopical gut content analysis .............................................................................. 41 3.7 Molecular gut content analysis..................................................................................... 41 3.7.a Primer design........................................................................................................ 41 3.7.b Insects and primary predation feeding trial........................................................... 42 3.7.c Species specificity................................................................................................. 42 3.7.d DNA Protocols ...................................................................................................... 42 3.8 Carabid daily rhythms .................................................................................................. 43 3.9 Carabid density in olive grove...................................................................................... 44 3.10 Simulation of carabid predation on B. oleae pupae and larvae................................. 44 4. RESULTS........................................................................................................................... 45 4.1 Preliminary test ............................................................................................................ 45 4.2 Quantitative tests ......................................................................................................... 52 4.2.a Behavioural analysis ............................................................................................. 52 4.2.b Food preferences.................................................................................................. 54 4.3 Microscopical gut content analysis .............................................................................. 60 4.4 Molecular gut content analysis..................................................................................... 61 4.5 Carabid daily rhythms .................................................................................................. 63 4.6 Carabid density in olive grove...................................................................................... 67 4.7 Simulation of carabid predation on B. oleae pupae and larvae................................... 69 5. DISCUSSION..................................................................................................................... 70 5.1 Preliminary test ............................................................................................................ 70 6.2 Quantitative tests ......................................................................................................... 71 6.3 Microscopical gut content analysis .............................................................................. 74 6.4 Molecular gut content analysis..................................................................................... 75 6.5 Carabid daily rhythms .................................................................................................. 75 6.6 Carabid density in olive grove...................................................................................... 76 6.7 Simulation of carabid predation on B. oleae pupae into the soil.................................. 78 7. CONCLUSION ................................................................................................................... 80 References............................................................................................................................. 83 _____________________________________________________________________________________ Rosalba Odoguardi PhD thesis 2 Acknowledgments ACKNOWLEDGMENTS I warmly thank my tutor, Prof. Tullia Zetto, for her essential support and consideration during my period of study, for her invaluable suggestions on manuscript organization, for linguistic revision and human prop; Prof. Pietro Brandamayr for structuring my research program and for many ideas; Dr. Teresa Bonacci for field and laboratory help and numerous other forms of entomological supports, for encouraging me to continue my work on behavioural research, Dr. Antonio Mazzei for helping me during sampling in experimental sites and carabid beetles identification, Dr. Anita Giglio and Dr. Federica F. Talarico for their suggestions during histological preparations and photographical acquisitions; Dr. Alessandro Massolo, for his useful support in statistical analyses. I am particularly grateful to Prof. Beatrice Bitonti, Dr. Leonardo Bruno for taking time to teach me DNA analysis laboratory methods. I thank Dr. Nino Iannotta, C.R.A, and his co-worker, in particular Dr. Stefano Scalercio, for research facilities, and all the other