INFLUENCE OF CLIMATIC VARIABLES AND BIOLOGICAL CONTROL AGENTS (NEMATODES) ON THE DISTRIBUTION AND SURVIVAL OF THE COFFEE BERRY BORER, HYPOTHENEMUS HAMPEI, IN PUERTO RICO By José Miguel García Peña A dissertation submitted to the DEPARTMENT OF BIOLOGY FACULTY OF NATURAL SCIENCES UNIVERSITY OF PUERTO RICO RÍO PIEDRAS CAMPUS In partial fulfillment of the requirements for the degree of DOCTOR IN PHILOSOPHY December 2019 San Juan, Puerto Rico © José Miguel García Peña All rights reserved ii This dissertation has been accepted by faculty of the: DEPARTMENT OF BIOLOGY FACULTY OF NATURAL SCIENCES UNIVERSITY OF PUERTO RICO RÍO PIEDRAS CAMPUS In partial fulfillment of the requirements for the degree of DOCTOR IN PHILOSOPHY Dissertation Committee: _________________________________________ José Carlos Verle Rodríguez, Ph.D., Advisor __________________________________________ William Gould, Ph.D. __________________________________________ Paul Bayman, Ph.D. __________________________________________ Alberto M. Sabat, Ph.D. __________________________________________ Miguel Acevedo, Ph.D. iii TABLE OF CONTENS LIST OF TABLES ................................................................................................................................vi LIST OF FIGURES ............................................................................................................................. vii ABSTRACT ...................................................................................................................................... viii DEDICATION .................................................................................................................................... x ACKNOWLEDGEMENTS .................................................................................................................... xi Chapter I GENERAL INTRODUCTION ................................................................................................. 1 1.1. Introduction ..................................................................................................................... 2 1.2. Coffee Berry Borer ............................................................................................................ 2 1.3. Biological Control ............................................................................................................. 3 1.4. Species Distribution Model ............................................................................................... 4 1.5. Entomopathogenic Nematodes ........................................................................................ 5 1.6. Environmental factors affecting nematode-insect interactions ......................................... 7 1.7. EPNs, native vs exotic ....................................................................................................... 7 1.8. Structure of this Dissertation ............................................................................................ 8 1.9. References ....................................................................................................................... 9 CHAPTER II Modelling Distribution of Suitable Coffee Berry Borer Habitat in Puerto Rico ............... 16 Abstract ..................................................................................................................................... 17 2.1. Introduction ................................................................................................................... 18 2.2. Material and Methods .................................................................................................... 19 2.2.1. Occurrence and Environmental Data ....................................................................... 19 2.2.2. Model ..................................................................................................................... 19 2.2.3. Historical temperature and precipitation ................................................................ 20 2.2.4. Relationship with Percent of infestation.................................................................. 21 2.3. Result ............................................................................................................................. 21 2.4. Discussion ...................................................................................................................... 23 2.5. Conclusion ...................................................................................................................... 25 2.6. Reference ....................................................................................................................... 25 2.7. Chapter II Tables ............................................................................................................. 30 2.8. Chapter II Figures............................................................................................................ 32 CHAPTER III Isolation and identification of entomopathogenic nematodes in Puerto Rico coffee growth zone................................................................................................................................... 37 Abstract ..................................................................................................................................... 38 3.1. Introduction ................................................................................................................... 39 iv 3.2. Methodology .................................................................................................................. 41 3.2.1. Soil samples ............................................................................................................ 41 3.2.2. Entomopathogenic nematode isolation................................................................... 41 3.2.3. Morphometric identification ................................................................................... 42 3.2.4. Molecular identification of EPN ............................................................................... 43 3.2.5. Statistical analysis ................................................................................................... 46 3.3. Results ............................................................................................................................ 46 3.4. Discussion ...................................................................................................................... 47 3.5. Conclusion and recommendation ................................................................................... 51 3.6. References ..................................................................................................................... 51 3.7. Chapter III Table ............................................................................................................. 57 3.8. Chapter III Figures........................................................................................................... 57 3.9. Chapter III Appendix ....................................................................................................... 61 Chapter IV Occurrence of Oscheius myriophila (Rhabditidae: Rhabditida) in Puerto Rico Coffee growing area. ................................................................................................................................ 64 Abstract ..................................................................................................................................... 65 4.1. Introduction ................................................................................................................... 66 4.2. Methodology .................................................................................................................. 67 4.2.1. Soil samples ............................................................................................................ 68 4.2.2. Nematodes ............................................................................................................. 68 4.2.3. Physical and chemical characteristics of soil ............................................................ 68 4.2.4. Statistical analysis ................................................................................................... 69 4.3. Results ............................................................................................................................ 69 4.4. Discussion ...................................................................................................................... 70 4.5. Conclusion ...................................................................................................................... 72 4.6. References ..................................................................................................................... 72 4.7. Chapter IV Tables ........................................................................................................... 77 4.8. Chapter IV Figures .......................................................................................................... 79 CHAPTER V GENERAL CONCLUSIONS AND RECOMMENDATIONS ................................................... 81 v LIST OF TABLES Chapter II Table 1. The environmental variables and altitude used to predict the suitable of Coffee berry borer habitat. ................................................................................................ 30 Chapter II Table 2. Relative contributions of the environmental variables to