INFORMATION TO USERS This manuscript has been reproduced from the microfilm master. UMI films the text directly from the original or copy submitted. Thus, some thesis and dissertation copies are in typewriter face, while others may be from any type of computer printer. The quality of this reproduction is dependent upon the quality of the copy submitted. Broken or indistinct print, colored or poor quality illustrations and photographs, print bleedthrough, substandard margins, and improper alignment can adversely afreet reproduction. In the unlikely event that the author did not send UMI a complete manuscript and there are missing pages, these will be noted. Also, if unauthorized copyright material had to be removed, a note will indicate the deletion. Oversize materials (e.g., maps, drawings, charts) are reproduced by sectioning the original, beginning at the upper left-hand corner and continuing from left to right in equal sections with small overlaps. Each original is also photographed in one exposure and is included in reduced form at the back of the book. Photographs included in the original manuscript have been reproduced xerographically in this copy. Higher quality 6M x 9" black and white photographic prints are available for any photographs or illustrations appearing in this copy for an additional charge. Contact UMI directly to order. University Microfilms International A Bell & Howell Information Com pany 300 North Z eeb Road Ann Arbor Ml 48106-1346 USA 313 761-4700 800 521-0600 Order Number 9326670 Physiological and behavioral interactions between the ectoparasitoidNasonia vitripennis (Walker) (Hymenoptera: Pteromalidae) and its pupal hostSarcophaga buliata P arker (Diptera: Sarcophagidae) Rivers, David Bradley, Ph.D. The Ohio State University, 1993 UMI 300 N. Zeeb Rd. Ann Aibor, Ml 48106 PHYSIOLOGICAL AND BEHAVIORAL INTERACTIONS BETWEEN THE ECTOPARASITOID NASONIA VITRIPENNIS (WALKER) (HYMENOPTERA: PTEROMALIDAE) AND ITS PUPAL HOST SARCOPHAGA BULLATA PARKER (DIPTERA: SARCOPHAGIDAE) DISSERTATION Presented in Partial Fulfillment of the Requirements for the Degree Doctor of Philosophy in the Graduate School of The Ohio State University By David Bradley Rivers, B.S. ***** The Ohio State University 1993 Dissertation Committee: D.L. Denlinger Approved by W.F. Hink R.W. Hall ^ m a m i - D.C. Smith Advisor ^ Department of Entomology To Dr. Harold L. Zimmack, who taught me about life. I also dedicate this dissertation to my family for their enduring support and encouragement. ii ACKNOWLEDGMENTS I express my sincere appreciation to Dr. David L. Denlinger for his guidance and encouragement throughout this research. I also thank the other members of my advisory committee, Drs. w. Fred Hink, Richard W. Hall, and D. Courtney Smith, for their insight and comments. Gratitude is extended to Jay Yoder, Brian Almond, Mei- Ling Zhang, Sherry Johnson, Karl Joplin, and George Yocum for many helpful comments throughout this study. I am also indebted to George Keeney for his insight and friendship, to Foster Purrington for his many helpful suggestions, and to the graduate students in the Department of Entomology for their friendship and support, in particular Dave Kallander, Bob Hancock, Diana Sammataro, Amadou Ba, and Jane Zumwalt. To my wife, Cheryl, I offer sincere thanks for your unshakable faith in me and for the love and encouragement you provided throughout my endeavors. I also thank my daughter, Megan, for giving me inspiration. VITA September 17, 1966 ............. Born - Kokomo, Indiana 1989 ................................ B.S., Ball State University, Muncie, Indiana 1989-1990 ......................... Research Assistant, Department of Biochemistry, The Ohio State University, Columbus, Ohio 1990-Present ....................... Teaching Assistant, Department of Entomology, The Ohio State Univeristy, Columbus, Ohio PUBLICATIONS Rivers, D. B. , C. N. Vann, and H. L. Zimmack. 1988. Specificity of the Bacillus thurinaiensis delta- endotoxins toward the larvae of Ostrinia nubilalis. Proceedings of the Indiana Academy of Science 98:21-24. Milne, R., A. Ge, D. Rivers, and D. Dean. 1990. Specificity of insecticidal crystal proteins: Implications for Industrial Standardization. In: Analytical Chemistry of Bacillus thurinaiensis. L. Hickle and W. Fitch (Eds), American Chemical Society, pp. 22-35. Rivers, D. B., C. N. Vann, H. L. Zimmack, and D. H. Dean. 1991. Mosquitocidal Activity of Bacillus laterosporus. Journal of Invertebrate Pathology 58:444-447. iv Ge, A., D. B. Rivers, R. Milne and D. H. Dean. 1991. Functional domains of Bacillus thurinoiensis insecticidal crystal proteins. Journal of Biological Chemistry 266(27):17954-17958. Rivers, D. B., W. F. Hink, and D. L. Denlinger. 1993. Toxicity of the venom from Nasonia vitripennis (Hymenoptera: Pteromalidae) toward fly hosts, nontarget insects, different developmental stages, and cultured insect cells. Toxicon (In press). FIELDS OF STUDY Major Field: Entomology (Insect Physiology) v TABLE OF CONTENTS DEDICATION .............................................. ii ACKNOWLEDGMENTS .......................................... iii VITA ..................................................... iv LIST OF TABLES ........................................... viii LIST OF FIGURES ....................................... ix INTRODUCTION ............................................ 1 CHAPTER PAGE I. HOST QUALITY AND ITS IMPACT ON FECUNDITY( DEVELOPMENTAL TIME, AND SEX RATIO IN NASONIA VITRIPENNIS. A GREGARIOUS, ECTOPARASITOID OF THE FLESH FLY, SARCOPHAGA B U L L A T A ............ 10 Introduction .............................. 10 Materials and Methods ..................... 11 R e s u l t s ..................................... 17 D i s c u s s i o n ................................ 36 II. DEVELOPMENTAL FATE OF THE FLESH FLY, SARCOPHAGA BULLATA. ENVENOMATED BY THE PUPAL ECTOPARASITOID, NASONIA VITRIPENNIS ............................ 44 Introduction .............................. 44 Materials and Methods ..................... 4 6 R e s u l t s ..................................... 51 D i s c u s s i o n ................................ 65 III. REDIRECTION OF METABOLISM IN THE FLESH FLY, SARCOPHAGA BULLATA. ENVENOMATED BY THE ECTOPARASITOID NASONIA VITRIPENNIS: A CONTRAST BETWEEN NONDIAPAUSING AND DIAPAUSING HOSTS . 70 Introduction .............................. 7 0 Materials and Methods ..................... 73 R e s u l t s ...................... 78 vi D i s c u s s i o n ................................ 90 IV. TOXICITY OF THE VENOM FROM NASONIA VITRIPENNIS (HYMENOPTERA: PTEROMALIDAE) TOWARD FLY HOSTS, NONTARGET INSECTS, DIFFERENT DEVELOPMENTAL STAGES, AND CULTURED INSECT CELLS ............. 97 Introduction.. .............................. 97 Materials and Methods ...................... 98 R e s u l t s ........................................ 103 D i s c u s s i o n ................................... 115 SUMMARY ................................................... 120 LIST OF REFERENCES .................................... 125 vii LIST OF TABLES TABLE PAGE 1. Fecundity of Jl. vitripennis on restricted body regions of its pupal host, S. bullata.............. 59 2. Injection of 20-hydroxyecdysone into pupae of S. bullata after envenomation ....................... 60 3. Hemolymph amino acid composition in pharate adults of S. bullata ................................. 85 4. Activity of the venom from Nasonia vitripennis toward natural and nontarget insect species. 110 5. Response of Trichoolusia ni (TN-368) and Sarcophaaa perearina (NIH SaPe4) cell lines to the venom of Nasonia vitripennis ................ 112 LIST OF FIGURES FIGURE PAGE 1. (A) Mean + SEM number of M. vitripennis adults produced per host on 4 species of fly pupae (£. bullata. £. crassipalpis. £. abnormis. and M- domestical (F« 15.68; df« 3,139; P < 0.01). (B) Mean + SEM time (days) required for 1*. vitripennis to develop from egg (deposition) to adult (emergence from host puparia) at 25°C, L15:D9 on pupae from each species of host (F= 117.1; df= 3,45; P < 0.01). 36-64 pupae of each fly host were used for each experiment. Columns preceded by the same letter do not differ significantly at P > 0.05 (ANOVA) . 28 2. The relationship between the number of adults of N . vitripennis produced and pupal weight (mg) of the host, S . bullata. The regress ion i s significant (y= 25*94 + 0.23x, P < 0.05, r2= 0.73, SEjg® 0.18) up to a host weight of 140 mg. Increases in host weight beyond 14 0 mg did not result in an increase in the number of parasitoids produced.............................. 3. (A) Mean + SEM number of eggs deposited (-•- host-fed and unfed wasps) and adults of N. vitripennis produced (-o- by host-fed and -□- unfed wasps) by host-fed and unfed wasps. (B) Adult eclosion (%) of JJ. vitripennis from envenomated (-•- host-fed and -G- unfed wasps) and unenvenomated (-•- host-fed, -o- unfed), nondiapausing puparia of S. bullata. (C) Mean ± SEM number of adult parasitoids produced on hosts of £. bullata killed by a 1 h exposure to —7 0°C. (D) Adult eclosion (%) of N. vitripennis resulting from egg transplantion to unenvenomated, dead hosts. (E) Oviposition and progeny production -o- by £. vitripennis on diapausing pupae of £. bullata. (F) Adult eclosion (%) of N. vitripennis resulting from egg transfer to unenvenomated, diapausing hosts. These experiments were replicated twice using ix 2 3-46 puparia in each replicate for each age and physiological state of the host ................. 32 4. (A) The proportion of males produced by host-fed -•- and