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ZEEB ROAD, ANN ARBOR, Ml 48106 18 BEDFORD ROW, LONDON WC1 R 4EJ, ENGLAND 8015874 F a u s t in i, D a r y l L . CHARACTERIZATION OF AN AGGREGATION PHER0M0NE AND ITS SITE OF PRODUCTION IN TRIBOLIUM CASTANEUM (HERBST) (COLEOPTERA: TENEBRIONIDAE) WITH COMPARATIVE NOTES ON ANALOGOUS STRUCTURES IN OTHER COLEOPTERA FAMILIES. The Ohio State University Ph.D . 1980 University Microfilms 300 N. Zeeb Road, Ann Arbor, MI 48106 18 Bedford Row, London WC1R 4EJ, England PLEASE NOTE: In all cases this material has been filmed in the best possible way from the available copy. Problems encountered with this document have been identified here with a check mark . 1. Glossy photographs 2. Colored illustrations 3. Photographs with dark background '4. Illustrations are poor copy_____ 5. Drin t shows through as there is text on both sides of page__________ 6. Indis^tnict, broken or small print on several pages throughout 7. Tightly bound copy with print lost in spine_________ 8. Computer printout pages with indistinct print 9. Page(s) lacking when material received, and not available from school or author_________ 10. Page(s) _________seem to be missing in numbering only as text follows _________ 11. Poor carbon copy_________ 12. Not original copy, several pages with blurred type 13. Appendix pages are poor copy_________ 14. Original copy with light type_________ 15. Curling and wrinkled pages_________ 16. Other University M icrd fiim s intemanonai Characterization of an Aggregation Pheromone and its Site of Production in Tvibol-ium oastaneum (Herbst) (Coleoptera: Tenebrionidae) with Comparative Notes on Analogous Structures in Other Coleoptera Families. DISSERTATION Presented in Partial Fulfillment of the Requirements for the Degree Doctor of Philosophy in the Graduate School of The Ohio State University By D. L. Faustini, A.A., B.A., M.S. * * * The Ohio State University 1980 Reading Committee: Approved By C. A. Triplehorn M. E. Clay Adviser D. L. Denlinger Department of Entomology DEDICATION I wish to dedicate this work in honor of ray parents John Mario and Evelyn Elizabeth Faustini and to my sisters, Alyce Dorner, Madonna Battaglia and Elayne Miller. ii ACKNOWLEDGEMENTS I wish to thank my major professor, Charles A. Triplehorn, for his assistance and support in this work, and for his warm friendship. I would also like to thank Dr. James E. Sargent who generously offered valuable advice and assistance with problems relating to this study. I am grateful to Dr. A. C. Waldron whose laboratory most of this work was performed. Special thanks are due to David W. Stutes and Dr. Thomas N. Taylor, Scanning Electron Microscopy Laboratory, Columbus, Ohio for facilities and technical assistance which allowed for the SEM micrographs in this study. Biological assays with the flour beetle were made possible by a travel grant from the Ohio State University Graduate School and courtesy of the Stored Products and Household Insects Laboratory, USDA, Department of Entomology, University of Wisconsin, Madison. I particularly wish to thank Dr. Wendell E. Burkholder, professor of Entomology at the above laboratory, for helping to arrange the visit, and whose laboratory the work was done. Finally, I am most grateful to my loving family, my assistants, Erik and Heidi and dearest wife Vicki for their many personal sacrifices during this study. iii VITA February 22, 1950 . Born - Cleveland, Ohio 1973.................. A.A., Mount San Antonio College, Walnut, California 197^.................. B.A., California State College, San Bernardino, California 197^-1977 ........... Research Assistant, Stored Grain Laboratory, California State College, San Bernardino, California 1976.................. M.Sc., California State College, San Bernardino, California 1979.................. Teaching Associate, Department of Entomology, The Ohio State Univer­ sity, Columbus, Ohio 1979-1980 ............ Research Assistant, Insect Physiology Laboratory, The Ohio State University, Columbus, Ohio PUBLICATIONS Faustini, D. L. (1976). The effect of sex and irradiation on crossing-over in Tribolium castaneum (Herbst). Tribolium Inform. Bulletin 19:88-89. Sokoloff, A., D. L. Faustini, M. A. Sokoloff and E. A. Sokoloff (1977). Observations of a natural population of Tribolium brevicornis LeConte. Tribolium Inform. Bulletin 20:135-138. Faustini, D. L. (1980). The ultrastructure of a male sexual dimorphic character in Tribolium castaneum (Herbst). Tribolium Inform. Bulletin {in -press). iv PROFESSIONAL PAPERS The effect of sex and irradiation on crossing-over in Tribolium castaneum. Biological Sciences Undergraduate Research Conference, Santa Clara, California. 1977- The morphology of a sexual dimorphic character in Tribolium oastaneum (Herbst) (Coleoptera: Tenebrionidae). Annual Meeting of the Entomological Society of America, Denver, Colorado. 1979. AWARDS Graduate Student Alumni Research Award, The Ohio State University, 1979- FIELDS OF STUDY Major Field: Functional Morphology and Behavior of Stored Grain Insects Studies in morphology: Professor Charles A. Triplehorn Studies in pheromone behavior: Professor Wendell E. Burkholder. v TABLE OF CONTENTS Page ACKNOWLEDGEMENTS iii VITA iv LIST OF TABLES viii LIST OF FIGURES x LIST OF PLATES xi INTRODUCTION ............................................ 1 GENERAL LITERATURE REVIEW.............................. 5 Tribolium spp....................................... 5 Pheromones.......................................... 8 GENERAL EXPERIMENTAL PROCEDURES....................... 12 Insect Rearing..................................... 12 Handling Pupae..................................... 15 Handling Adults ................................... 16 IDENTIFICATION OF THE EXOCRINE GLAND ................ 17 Introduction . ................................... 17 Morphology of the Male Setiferous Puncture . 23 Scanning Electron Microscopy .............. 23 Solubility Properties of the Globular Secretion . .............................. 23 Ultrastructure of the Setiferous Puncture . 26 Secretion Accumulation with Beetle Age . 28 Histology..................................... 28 Discussion.......................................... 42 LABORATORY BIOASSAY AND CHEMICAL ANALYSES ............ 47 Introduction................................ 47 Materials and Methods ............................ 49 vi TABLE OF CONTENTS (CONTINUED) Page Handling of Insects......................... 49 Olfactometer and Laboratory Bioassay Conditions................................ 50 Multiple-Choice Olfactometer .............. 50 Isolation of the Male Setiferous Secretion................................... 52 Chemical Analyses............................ 52 Gas Chromatography - packed column. 5 3 Gas Chromatography - glass capillary column................................ 53 Experimental Results.............................. 5^ Female and Male Behavioral Response .... 5^ Multiple-Choice Olfactometer Bioassay . 5^ Female Response .............. 5^ Male Respo n s e ......................... 59 Analysis of the Secretion.................. 76 Discussion.......................................... 8l ANALOGOUS SEXUAL DIMORPHIC MALE SETIFEROUS STRUCTURES 86 Introduction....................................... 86 Materials and Methods ............................ 87 Scanning Electron Microscopy .............. 87 I n s e c t s ..................................... 88 Experimental Results.............................. 88 Tenebrionoidea .............................. 88 Staphylinoidea....................... 101 Bostrichoidea................................... 101 Dermestoidea ................................ 108 Cleroidea........................................108