2004 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? Biting and filth breeding flies transmit bacterial, viral, protozoan, and filarial disease agents to animals and humans. Annoyance and irritation from mosquito and filth breeding fly bites impedes agricultural productivity, economic development, and the use of recreational areas. Effective general and specific surveillance traps and new trap technologies are needed to provide early warning of vector borne diseases that can trigger specific and appropriately timed control responses to protect animals and humans from arthropod attack and the transmission of arthropod-borne disease agents. To meet this need, ARS-CMAVE scientists in the Mosquito and Fly Research Unit develop new surveillance tools that can fit into an integrated pest abatement system to safely and effectively manage biting and filth-breeding flies. Over one million pounds (active ingredient) of pesticide are used each year in the USA to protect animals and humans from biting and filth inhabiting insects. These pesticides are the only tool available for emergency vector control and are the mainstay of conventional integrated pest management (IPM) systems in animal and public health. Within the next decade, however, pesticide use will be severely restricted by ecological, safety, legal, and economic factors. Better surveillance tools and alternative control technologies for biting and filth inhabiting arthropods are needed now to maintain our current pesticide-based capacity for control of these pests and disease vectors in the future. This research is entirely (100%) within the scope of National Program 104, Veterinary, Medical, and Urban Entomology Research. The CRIS addresses the following National Program Action Plan Goals: 1.1 Identify Aspects of Arthropod Behavior Vulnerable to Control (1.1.1 & 1.1.2); 2.2 Surveillance (2.2.1; 2.2.3; & 2.2.4); 3.1 Genomics and Host-Pathogen Interaction (3.1.3); 3.2 Neural, Sensory, and Reproductive Biology (3.2.1 & 3.2.2). These goals represent efforts to provide new and improved surveillance tools that together with non-pesticidal, biologically-based control methods would provide safe and effective control systems in urban, agricultural and natural ecosystems for these serious medical, veterinary, and agricultural pests. Thus, the research problems addressed by this CRIS support the Pesticide Reduction Act through the development of integrated pest management technologies, as well as the Food Quality Protection Act of 1996 through the development of reduced-risk control strategies. Collaborators in this research include the Armed Forces Pest Management Board of the Department of Defense, the U. S. Agency for International Development, the World Health Organization, and the International Atomic Energy Agency. 2.List the milestones (indicators of progress) from your Project Plan. Milestone 1 (24 months) - Characterize mating attractants in Musca subspecies including presumptive sex pheromones. Characterize cuticular hydrocarbons of muscid fly pupae as presumptive pupal kairomones for hymenopterous parasitoids. Documentation: peer-reviewed scientific publications/products, i.e., patents, detection, surveillance, and control technology for biting and filth breeding flies. Milestone 2 (36 months) - Isolate, identify, and evaluate human, plant, and animal volatiles, and volatiles from larval developmental sites, as feeding and oviposition attractants for mosquitoes and other biting flies. Design, develop, and test field olfactometer systems to measure attraction responses in natural populations of mosquitoes and other biting flies. Documentation: peer-reviewed scientific publications/products, i.e., patents, detection, surveillance, and control technology for biting and filth breeding flies. Milestone 3 (48 months) - Utilize known biting fly attractants, new attractants, and known, or new, configurations of trapping apparatus and/or attractive 'targets' to develop novel monitoring and surveillance technology. Documentation: peer-reviewed scientific publications/products, i.e., patents, detection, and surveillance technology for biting and filth breeding flies. Milestone 4 (48 months) - Identify, synthesize, and test candidate compounds as attractants for use in baits for fly control and or for use in the management of hymenopterous parasitoids as biological control agents for filth breeding flies. Identify and test aggregation pheromones and biologically active natural oils for litter beetle control. Documentation: peer-reviewed scientific publications/products, i.e., patents, detection, and surveillance technology for biting and filth breeding flies. 3.Milestones: A. List the milestones that were scheduled to be addressed in FY 2004. How many milestones did you fully or substantially meet in FY 2004 and indicate which ones were not fully or substantially met, briefly explain why not, and your plans to do so. Milestone 3 (48 months) - Utilize known biting fly attractants, new attractants, and known, or new, configurations of trapping apparatus and/or attractive 'targets' to develop novel monitoring and surveillance technology. Documentation: peer-reviewed scientific publications/products, i.e., patents, detection, and surveillance technology for biting and filth breeding flies. Milestone 4 (48 months) Identify, synthesize, and test candidate compounds as attractants for use in baits for fly control and or for use in the management of hymenopterous parasitoids as biological control agents for filth breeding flies. Identify and test aggregation pheromones and biologically active natural oils for litter beetle control. Documentation: peer-reviewed scientific publications/products, i.e., patents, detection, and surveillance technology for biting and filth breeding flies. The Mosquito & Fly Unit has undergone CRIS contraction and expansion within the last two years. The Milestones were taken from the original Project Plan (due to expire February 2005) and segregated by topic into the current two CRIS projects (6615-32000-039-00D, Control of and Protection from Biting and Filth Breeding Insects, and 6615-32000-040-00D, Surveillance and Ecology of Mosquito, Biting and Filth Breding Insects. All milestones have been either fully accomplished (Milestone. 3)or partially accomplished (Milestone 4). New emphasis on novel surveillance tools, GIS, and decision systems is evident in our new Project Plan that is currently under OSQR Review. B: List the milestones that you expect to address over the next 3 years (FY2005, 2006, 2007). What do you expect to accomplish, year by year, over the next 3 years under each milestone? This Project Plan will terminate February 2005; therefore, the Project Plan that is currently under review will cover future work. 4.What were the most significant accomplishments this past year? A. Single most significant accomplishment during FY 2004: Chemical Attractant for Mosquitoes. Lactic acid and octenol were shown to effectively attract Aedes albopictus (Asian Tiger mosquito). This mosquito species is a serious nuisance to people during the daylight hours and is a prime suspect as a species that transmits West Nile virus to humans. Unit Scientists discovered that the combination of attractants was most effective in attracting this species to the trap and resulted in our highest captures. This is important because improved surveillance methods are needed for this difficult to trap species that can trigger appropriately timed control efforts. B: Other Significant Accomplishments: Volatiles from Blood Attract Mosquitoes. Blood is a known attractant of mosquitoes and other blood-feeding flies. Unit Scientists collected and identified volatiles from bovine and avian blood. The range of compounds was assayed in olfactometer and cage assays for attraction and the landing responses of Aedes and Culex mosquitoes. There were significant differences in responses between mosquito species and between behaviors examined. This is the first critical step for development of Culex specific attractants that can be used as lures in a trapping system. These target-specific traps will enhance population and arbovirus surveillance for West Nile virus. Visual Attractants for House Flies. The attractiveness of various visual traps to house flies and stable flies was assessed in the field. Clear Alsynite was the most attractive material for both species of flies, followed by Sunbrella Royal Blue fabric. Targets consisting of alternating patterns of blue fabric and clear Alsynite were more attractive than predicted based on single-color testing, indicating synergism due to color contrasts. Visual targets, in combination with a killing agent, could be used to intercept flies that would otherwise emigrate from farms and invade the homes of neighbors. C. Significant activities that support special target populations: None. D. Progress report: None. 5.Describe the major accomplishments over the life of the project, including their predicted or actual impact. (1) New and improved traps facilitated the accurate and efficient detection, population monitoring, and control of mosquito vectors and other blood sucking and filth inhabiting arthropods. (2) Development of molecular and morphological diagnostic tools for mosquitoes facilitated the accurate identification of disease vectors. (3) The discovery of human-produced chemical attractants for mosquitoes. Attractants are being used in trapping systems for the control of mosquito vectors of exotic and indigenous disease agents. This research provides for the protection of animals and humans from blood sucking and filth breeding arthropods and the disease agents they transmit. 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? New mosquito surveillance technology was presented to mosquito control workers for northeastern Florida at a workshop held at Anastasia Mosquito Abatement District. Invited to participate in an afternoon demonstration of new and currently used traps for nuisance flies and mosquitoes at the Department of Defense Pest Management Workshop, Jacksonville Naval Air Station, Jacksonville, FL, February 9-11, 2004. Surveillance techniques for adult and immature stages of Culicoides biting midges were demonstrated to personnel from Vector Disease Control and ADAPCO (Orlando, FL). They are currently using this technology in projects being conducted in the Caribbean Basin. New mosquito surveillance technology was presented to mosquito control workers for northeastern Florida at a workshop held at Anastasia Mosquito Abatement District. 7.List your most important publications in the popular press and presentations to organizations and articles written about your work. Panorama Magazine (Italian) interviewed Dr. Bernier for an article entitled "Ronzii Precoci L'incubo E Nell 'Aria Rieccole", April 29, 2004 issue. Dr. Bernier was interviewed for an article that appeared in the Colorado Springs Gazette, "Mosquitoes seem to enjoy feasting on you but leave your next-door neighbor alone." July 12, 2004. Dr. Kline's work on mosquito surveillance appeared in an article, "Pest Control", in the July issue of Men's Health. Dr. Kline's work on mosquito attractants led to an article by Dean Traiger, entitled "How Attractive Are You?" in the July issue of Today's Cacher. Surveillance CRIS scientists presented the following papers at the International Congress of Entomology, held in Brisbane, Australia: "Chemical analysis and laboratory bioassays of host odors to identified compounds that attract or inhibit the attraction of mosquitoes"; "Evaluation of volatile compounds from blood as attractants for Aedes and Culex mosquitoes"; and "Novel mosquito traps; can they control mosquitoes? Review Publications Wilkerson, R.C., Reinert, J.F., Li, C. 2004. Ribosomal dna its2 sequences differentiate six species in the anopheles (anopheles) crucians complex (diptera: culicidae). Journal of Medical Entomology. 41(3):392-401. BERNIER, U.R., KLINE, D.L., POSEY, K.H., BOOTH, M.M., YOST, R.A., BARNARD, D.R. SYNERGISTIC ATTRACTION OF AEDES AEGYPTI (L.) TO BINARY BLENDS OF L-LACTIC ACID AND ACETONE, DICHLOROMETHANE, OR DIMETHYL DISULFIDE. JOURNAL OF MEDICAL ENTOMOLOGY. 2003. v.40(5).p.653-565.