CLASSICAL BIOLOGICAL CONTROL OF MEDITERRANEAN FRUIT FLY, Ceratitis capitata (WIEDEMANN), (DIPTERA: TEPHRITIDAE): NATURAL ENEMY EXPLORATION AND NONTARGET TESTING A Dissertation by MARCIA KATHERINE TROSTLE DUKE Submitted to the Office of Graduate Studies of Texas A&M University in partial fulfillment of the requirements for the degree of DOCTOR OF PHILOSOPHY May 2005 Major Subject: Entomology CLASSICAL BIOLOGICAL CONTROL OF MEDITERRANEAN FRUIT FLY, Ceratitis capitata (WIEDEMANN), (DIPTERA: TEPHRITIDAE): NATURAL ENEMY EXPLORATION AND NONTARGET TESTING A Dissertation by MARCIA KATHERINE TROSTLE DUKE Submitted to Texas A&M University in partial fulfillment of the requirements for the degree of DOCTOR OF PHILOSOPHY Approved as to style and content by: ________________________ _________________________ Robert A. Wharton James B. Woolley (Chair of Committee) (Member) _______________________ _________________________ Marvin K. Harris Thomas L. Linton (Member) (Member) _______________________ Kevin Heinz (Head of Department) May 2005 Major Subject: Entomology iii ABSTRACT Classical Biological Control of Mediterranean Fruit Fly, Ceratitis capitata, (Wiedemann), (Diptera: Tephritidae): Natural Enemy Exploration and Nontarget Testing. (May 2005) Marcia Katherine Trostle Duke, B.S., Texas A&M University Chair of Advisory Committee: Dr. Robert A. Wharton This work covers stages one through seven (of nine stages) of a classical biological control program for Mediterranean fruit fly (=medfly), Ceratitis capitata (Wiedemann). Major research objectives concentrate on stage five (exploration and collection of natural enemies), and stage seven (testing and selecting natural enemies for additional work). Coffee was collected monthly from three locations in Kenya from November 1997 through July 1999. Four species of tephritid flies and ten parasitoid species were recovered. Four guilds of parasitoids were recorded, and two egg-prepupal endoparasitoids, Fopius caudatus (Szépligeti) and F. ceratitivorus (Wharton), were discovered. The oviposition behavior of these two species is contrasted. Domination of this tropical parasitoid assemblage by koinobionts is discussed relative to the dominance of temperate fruit-infesting tephritid systems by idiobionts. Fruit handling procedures were examined for impact on overall percent emergence and specifically percent emergence of flies versus parasitoids. It was determined that stirring samples had a significant positive effect on overall emergence, however daily misting of fruit did not. The only treatment without a significant bias in fly emergence over parasitoids was the stirred/dry treatment. Effects of these results on rearing procedures are discussed. Host specificity and host suitability of parasitoids reared from coffee were examined via: (1) association of parasitoids with host flies based on characteristics of the fly puparia from iv which parasitoids emerged, (2) rearing of cucurbit infesting tephritids and their parasitoids in Kenya, (3) rearing of flowerhead infesting tephritids and their parasitoids in Kenya and Hawaii, and (4) host range testing of Psyttalia species in Kenya and Hawaii. These results are discussed in terms of their utility for predicting nontarget effects. Psyttalia concolor (Szépligeti) was shipped to Hawaii and tested against the nontarget gall forming tephritid Procecidochares utilis Stone introduced to control the weed Ageretina adenophora (Maui pamakani). Psyttalia concolor failed to attack the gall-forming P. utilis both in choice and no-choice tests, but readily attacked tephritid larvae offered in fruit in choice tests. Recommendations for further testing and release of the parasitoids from Kenya are discussed for Hawaii and Latin America. v To my parents, Mark and Susan, who have supported me unconditionally and endured 30 years of questions, answering when necessary and sending me on my way when it was mine to figure out alone. Amazingly never getting angry; though once in a while asking for a short five minutes of quiet. To my new husband, Thadeus, may he have the patience to support me and endure my questions for the rest of his lifetime. To my best friend, Sunny Ruth, DVM, this would have not been possible without your friendship, love and sacrifice. And to the Lord, Jesus looked at them and said, “With man this is impossible, but with God all things are possible.” Matthew 19:26 vi ACKNOWLEDGEMENTS The work in this dissertation could not have been accomplished without the support and guidance of my professors, family and friends. I am extremely grateful to have been mentored by Dr. Wharton as my major professor. I thank him for his patience and support during the long and arduous process of completing my research and writing the results. I am glad he recruited me to such a tough project and never gave up on me when times got hard. I thank him for teaching me to be a scientist, the value of focus, and the important lesson that the answer to the big questions can only be found by answering lots and lots and lots of smaller questions. I know all he has taught me will provide me a solid foundation for answering the hard questions that will come in my future. In other words, when I am out in the real world, all I have to ask myself is, does the problem, solution or explanation pass the “Wharton test?” If the answer is yes, continue. If the answer is no, stop, rethink and reexamine the issue prior to proceeding forward. I also appreciate the rest of my committee members, Drs. Jim Woolley, Tom Linton, Marvin Harris and George Teetes. I especially thank Dr. Harris for replacing Dr. Teetes when the latter retired. Their service to my education has been invaluable. I am appreciative to many faculty members in the Department of Entomology for their assistance particularly Drs. Kevin Heinz, Larry Keeley and Ted Wilson. I am also indebted to the professors who have taught my courses and shared their knowledge. On my trips around the world I was offered considerable assistance. I thank the following people and organizations in Hawaii: Russell Messing and the University of Hawaii (U of H) for providing laboratory space; the Hawaii Department of Agriculture (HDOA) for allowing me work-space in the quarantine facility, Mohsen Ramadan and Ken Teramoto also both of HDOA; the Honolulu USDA/ARS Tropical Fruit, Vegetable and Ornamental Crop vii Research Laboratory in Manoa for fruit fly larvae, adult flies and yeast hydrolysate (special thanks to Danny Seo, and Brian Fujita at the USDA); Nathan Peabody, a graduate student at U of H who worked on the Psyttalia portion of the project; and Dr. Marshall Johnson for helping me feel at home in Hawaii. In Kenya there are also many to thank including: the International Centre for Insect Physiology and Ecology (ICIPE) for laboratory space and the Duduville guest house for a place to rest my weary head at night; the IFAD fruit fly rearing team, especially Peterson Nderitu, for rearing fruit fly larvae for experiments; Samira Mohamed for collegial discussions, hospitality and friendship; Bob Copeland and Bill Overholt for their guidance and hospitality in a strange land as well as contributions to my project; and Vincent Shitacke and John for technical assistance. I express thanks to the following people and groups in Guatemala: MOSCAMED Program “La Aurora” laboratory including Miguel Lopez and Gustavo Baeza, USDA-APHIS- PPQ-CPHST Guatemala group including Pedro Rendon and the staff at the USDA-APHIS/ MOSCAMED quarantine facility in San Miguel Petapa where I was provided space to conduct my experiments. In addition I thank John Sivinski (USDA-ARS, CMAVE in Gainesville, FL) and Tim Holler (USDA-APHIS-PPQ-CPHST, Gainesville, FL) for their contributions to my research in Guatemala and Kenya as well as their hospitality. I also thank Ian White (International Institute of Entomology, London) for identification of the tephritid collected from Zinnia flowerheads in Kenya. The assistance of student workers was invaluable. I am forever grateful to Danielle McCain for the dissection of thousands of puparia as well as slide mounting mouthhooks and anterior and posterior spiracles for examination. From Hawaii, I thank Roxanne for helping viii conduct Psyttalia nontarget experiments in 2000. I also thank Petra Franzen for illustrations and William Wells for automontage images of mouthhooks, and anterior and posterior spiracles. This project was funded in part by grants to the Wharton Laboratory, particularly USDA/NRI grant no. 9703184, USAID grant no. PCE-G-00-98-00048-00, and USDA/CSREES/IFAFS grant no. 00-52103-9651 and in part by USDA/CSREES Special Grant no. 96-34135 and other funding from Russell Messing. I am grateful for the Wharton Lab including Matt Yoder, Matt Buffington (who identified the species of eucoline figitid from the Asteraceae flowerheads in Hawaii), Bob Kula, and most especially Amy Bader. She is a very special person and her friendship has been instrumental in my sanity. I also want to thank Jason Mottern for his friendship. He is an amazing young man with a golden heart and an excellent vice-president. Although it may seem strange, I thank all of the young men whom I dated during the years I worked on my project. Although they will never know it, and they will never see the glory of this document, I thank each of them for their support when they were an important part of my life. I also want to thank my good friends Matthew Hoddie and Elizabeth Bell for opening their house and providing me a place to stay when I needed to work on my dissertation and I no longer lived in College Station Finally, but definitely not subordinate to any of the above, I thank those closest to me. I have the most amazing parents who have put a lot of thought into how to raise a child. I hope in this dissertation they can see the fruits of their labor. I thank them for their sacrifices to my endeavors throughout my life. I would never have made it to the end of this project without all their love, the great value system they instilled in me, the never-ending interest in my progress, and the bubbling stream of enthusiasm convincing me the pain is worthy of the prize.