2005 Annual Report 1.What major problem or issue is being resolved and how are you resolving it (summarize project aims and objectives)? How serious is the problem? What does it matter? New chemical tools are urgently needed to protect humans from blood-sucking arthropods that can transmit diseases like West Nile virus, malaria, and leishmaniasis. Using a multidisciplinary approach involving entomology, insect behavior, vector biology, and organic, analytical and computational chemistry, the Chemicals Affecting Insect Behavior Laboratory (CAIBL) has developed novel bioassay systems in an attempt to identify new arthropod repellents and compounds having behavioral and/or insecticidal modes of action. In addition, new chemical attractants will be sought for improved mosquito surveillance, and spatial repellent compounds will be sought to guard against biting arthropods entering areas of human habitation. The ultimate goal is to provide safe and effective means to reduce risks of disease transmission to humans. This program falls within the National Program 104 Mission to develop effective means to prevent or suppress insects and other arthropods that affect human well being and to enhance the safety and quality of life for the U.S. public. 2.List the milestones (indicators of progress) from your Project Plan. Year 1 (FY2005) Insects rearing optimized using blood-membrane feeding. Standardized high through put screening (HTS) assay system in place. Computational methodology for compound selection from BASF library in place. Design and synthesis of 3 pyrethroids containing molecular spacers complete. Design and synthesis of several polymers with functional groups suitable for attaching repellent units studied. Candidate compounds bioassayed against a mosquito. Method for chemically attaching the modified pyrethroids to polymer studied. Known repellent compounds studied using Culex quinquifaciatus odorant binding proteins (OBPs) affinity columns. Preliminary single cell electrophysiological screening recording (SCR) experiments conducted on Culex pipiens. Cloning of OBP from Anopheles stephensi and Aedes aegypti initiated. Year 2 (FY2006) HTS-evaluated 3,000 BASF library compounds and two new model compounds identified. Computational chemistry models optimized. EPA requirements for product registration determined. Synthetic polymers containing chemically bonded pyrethroids obtained and bioassayed. Most promising polymers are spun into fibers. Affinity columns prepared using cloned Anopheles stephensi and Aedes aegypti OBPs and SCR started with the two species Year 3 (FY2007) Synthetic methods for preparation of model compounds for field tests developed. Field tests experimental designs complete. Fabric development studied. Complete experimental designs for field tests. Compound screening started using CquiOBP affinity columns. Year 4 (FY2008) Final evaluation studies of most promising compounds completed. New refined and efficient computational chemistry strategies identified and published. New fabrics developed. Complete SCR comparisons in three mosquito species. Year 5 (FY2009) EPA registration and technology transfer of new products in final stages. Finish field tests to test vector-control efficacy. Finish QSAR studies using synthetic model compounds having optimized OBP affinity. 4a.What was the single most significant accomplishment this past year? A walk-in mosquito rearing facility was developed to permit the rearing of thousands of mosquitoes for use in behavioral research and in a chemical screening program for discovery of new mosquito feeding deterrent compounds and knock down toxicants. 4b.List other significant accomplishments, if any. None. 4c.List any significant activities that support special target populations. None. 4d.Progress report. None. 5.Describe the major accomplishments over the life of the project, including their predicted or actual impact. This research project started 12/10/04 and replaced Project number 1275-22000-192-00D, MANIPULATION OF ARTHROPOD BEHAVIOR FOR PROTECTION OF LIVESTOCK AND HUMANS. 6.What science and/or technologies have been transferred and to whom? When is the science and/or technology likely to become available to the end-user (industry, farmer, other scientists)? What are the constraints, if known, to the adoption and durability of the technology products? None. It is anticipated it will take at approximately 5-10 years to deliver the technology to our customers that include the U. S. military, public health-care industries, and pest management industries. Constraints include the identification of new and safe repellents and registration requirements of the Environmental Protection Agency (EPA). 7.List your most important publications in the popular press and presentations to organizations and articles written about your work. (NOTE: List your peer reviewed publications below). "From the back yard to the bloodstream: The summer bug battle is rejoined" by Dennis Obrien, Baltimore Sun June 13, 2005. "If you go down to the woods today, rub some leaves on yourself" by Peter Calamai, Toronto Star July 2, 2005. Review Publications Chauhan, K.R., Klun, J.A., Debboun, M., Kramer, M. 2005. Feeding Deterrent Effects of Catnip Oil Components Compared with Two Synthetic Amides Against Aedes aegypti. Journal of Medical Entomology. 42:643-646. Norden, B.B., Webb, R.E., Mccardle, P.W., White, G.B. 2005. A new fixing technique using rice vinegar prior to mosquito dissection.. Journal of Kansas Entomological Society. 78:189-191. Klun, J.A., Kramer, M., Debboun, M. 2005. A new in vitro bioassay system for discovery of novel human-use mosquito repellents.. American Mosquito Control Association. 21:64-70. Cantrell, C.L., Klun, J.A., Duke, S.O., Bryson, C.T., Kobaisy, M.M. 2005. Isolation and identification of mosquito-repellent terpenoids from leaves of american (callicarpa americana) and japanese (callicarpa japonica) beautyberry. Journal of Agricultural and Food Chemistry. 53:5948-5953.