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ABSTRACT BISSINGER, BROOKE WITTING. Novel Methods of Hematophagous Arthropod Control. (Under the direction of R. Michael Roe). Ticks are important vectors of human and animal diseases. One protective measure against ticks is the use of personal arthropod repellents. Here, the history and efficacy of tick repellents, discovery of new repellents, and areas in need of attention such as assay methodology, repellent formulation, and the lack of information about the physiology of repellency are reviewed. Studies were conducted to examine the efficacy of the repellent BioUD with the active ingredient 7.75% 2-undecanone, originally derived from wild tomato plants. BioUD was compared with 7 and 15% deet using arm-in-cage studies against the mosquitoes Aedes aegypti and Ae. albopictus. No differences were found in mean repellency over 6 h after application between BioUD versus 7 and 15% deet for Ae albopictus. For Ae. aegypti, no differences were found over the same time period for 7% deet. Compared to 15% deet, BioUD was less repellent over the 6-h test period. Human subject field trials were conducted in North Carolina and Ontario comparing the repellency of BioUD to products containing 25 and 30% deet. BioUD provided the same repellency or was more efficacious than 25 and 30% deet, respectively. Repellent efficacy of BioUD and 98.1% deet against ticks was examined in the laboratory using a choice test between repellent-treated and control filter paper surfaces for Amblyomma americanum, Dermacentor variabilis and Ixodes scapularis. BioUD provided greater repellency against A. americanum and I. scapularis than deet. No difference was found between BioUD and deet against D. variabilis. In head-to-head assays between BioUD and deet, undiluted and 50% dilutions of BioUD were more repellent than undiluted deet against all three species. A 25% dilution of BioUD was more repellent than deet against A. americanum while no differences were found between a 25% dilution of BioUD and deet against D. variabilis and I. scapularis. Based on regression analysis, the concentration of BioUD required for equivalent repellency to 98.1% deet was 39.5% for D. variabilis and 29.7% for I. scapularis. A log-probit model could not be constructed for A. americanum from the dosages tested. Repellency of BioUD was compared to five repellents against A. americanum and D. variabilis in two-choice bioassays on treated versus untreated cotton cheesecloth. Overall mean percentage repellency against both species was greatest for and did not differ significantly between BioUD and products containing 98.1% deet, 19.6% IR3535, and 30% oil of lemon eucalyptus. Products containing 5 and 15% Picaridin and 0.5% permethrin were also repellent compared to untreated controls but to a lesser degree than BioUD. The four most active repellents were directly compared in head-to-head bioassays. BioUD provided significantly greater overall mean percentage repellency than IR3535 for A. americanum and D. variabilis. BioUD was significantly more repellent than oil of lemon eucalyptus for A. americanum but did not differ significantly in repellency against D. variabilis. No statistically significant difference in overall mean percentage repellency was found between BioUD and deet for A. americanum or D. variabilis. Laboratory trials were also conducted to determine the repellent activity of BioUD against D. variabilis on human skin. BioUD repelled ticks at least 2.5 h after application to human skin. Characterization of the expressed genes in the tick central nervous system could lead to a greater understanding of the control of development and reproduction at the molecular level. A transcriptome to the female D. variabilis synganglion identified 21,119 unique putative gene sequences, of which 7,379 had significant matches to the GenBank nonredundant database. Microarray analysis comparing synganglia from unfed, partially fed, and mated replete females revealed that 121 of these genes were differentially regulated. Novel Methods of Hematophagous Arthropod Control by Brooke Witting Bissinger A dissertation submitted to the Graduate Faculty of North Carolina State University in partial fulfillment of the requirements for the Degree of Doctor of Philosophy Entomology Raleigh, North Carolina 2009 APPROVED BY: _______________________________ ______________________________ R. Michael Roe Charles Apperson Committee Chair ________________________________ ________________________________ Christina Grozinger James Harper ________________________________ D. Wesley Watson DEDICATION This dissertation is dedicated to my dad, Thomas Joseph Witting. Thank you for all of the camping and fishing trips, hikes in the woods, gardening, lightening bugs in jars, and my first butterfly net. You taught me to appreciate and be in awe of the natural world. ii BIOGRAPHY Brooke Witting Bissinger was raised in Durham, NC. She attended Warren Wilson College in Swannanoa, NC and received a Bachelor of Arts in Environmental Studies in 2000. Brooke completed a Master of Science degree at North Carolina State University in the Department of Entomology under the direction of Drs. David Orr and Michael Linker in 2006. She then began work on her doctorate in the same department with Dr. R. Michael Roe. iii ACKNOWLEDGEMENTS A number of people played a vital role in the research that went into this dissertation, particularly the members of my graduate committee: Dr. Michael Roe, Dr. Charles Apperson, Dr. Christina Grozinger, Dr. Wes Watson, and Dr. James Harper. I am particularly grateful to Dr. Roe for fostering a positive working environment and encouraging teamwork and camaraderie among the students and postdoctoral scientists in his laboratory. We have all learned that coffee breaks are a good time to exchange ideas about science and a time to get to know one another so that we can build lifelong friendships and collaborations. Dr. Roe has taught us to think creatively and that every idea is worth pursuing. I extend sincere gratitude to Drs. Kevin Donohue and Sayed Khalil for mentoring me in the field of molecular biology. I would like to acknowledge Ana Cabrera for tirelessly spending hours dissecting out tick synganglia; and Luma Abu Ayyash, Nicholas Kimps, Jaap van Kretchmar, and Jiwei Zhu for assistance with repellency trials. I sincerely appreciate the guidance and generous supply of ticks provided by Dr. Daniel Sonenshine. I also appreciate Dr. Sonenshine allowing me to visit his lab on several occasions and for setting up experiments prior to my arrival. I am grateful to Drs. Cavell Brownie, Consuelo Arellano, and Christof Stumpf for help with statistical analyses and Drs. Chris Ashwell and Sarah Kocher for volunteering their time to teach me microarray analysis. Finally, I am grateful to my husband Michael Bissinger for supporting me in many ways throughout my degree program and for teaching me how to create better figures for my publications and presentations. iv TABLE OF CONTENTS LIST OF TABLES........................................................................................................... viii LIST OF FIGURES .............................................................................................................x TICK REPELLENTS: PAST, PRESENT, AND FUTURE ................................................1 Abstract................................................................................................................................2 Introduction..........................................................................................................................3 Sensory Perception...............................................................................................................5 Assay methods for tick repellency.......................................................................................7 The first synthetic repellents..............................................................................................11 DMP.......................................................................................................................12 Indalone..................................................................................................................13 Ethyl hexanediol ....................................................................................................14 6-2-2.......................................................................................................................14 Modern synthetic repellents..............................................................................................15 Deet........................................................................................................................15 Permethrin..............................................................................................................16 DEPA .....................................................................................................................19 Piperidines..............................................................................................................19 Plant-based repellents ............................................................................................21 Terpenoids..............................................................................................................22 Plant growth regulators..........................................................................................25 Anti-tick pasture plants..........................................................................................26
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