Location: Mosquito and Fly Research Unit
2006 Annual Report
This research is entirely (100%) within the scope of National Program 104, Veterinary, Medical, and Urban Entomology Research. This Project Plan addresses at least seven goals in the NP 104 Action Plan concerning mosquito and fly behavior, host-pathogen interactions, neural and sensory studies, repellents and biological control strategies including the specific goals: 1.1.1 Characterize the oviposition behavior of mosquitoes and midges and isolate environmental factors that attract or repel; 1.1.2 Determine the dispersal patterns, breeding habits and host attractions of horn flies, house flies and stable flies that may be useful in devising control strategies; 3.2.3 Develop measurements of electrophysiological activation for use in selecting vector repellents; 4.1.1 Develop the capacity to test existing compounds for toxicity against Diptera and ticks that transmit disease. Develop and use computational models for rapidly screening and predicting efficacy of available compounds from configural extrapolation. Devise formulations or combinations of compounds that can be used in regions anywhere in the world, without foreknowledge of resistance patterns; 4.1.2 Develop and test novel means of applying pesticides and repellents that are more efficient, inexpensive and selective, including the development of toxic baits, methods to bond agents to material, and area repellents; 4.1.3 Identify and test new classes of topical and area repellents from previously synthesized or natural volatiles. Develop carrier formulations; 4.2.1 Isolate, identify, cultivate, characterize and test natural pathogens and predators of vectors and pests. Develop methods to enhance the specificity and lethality of control agents; 4.3.1 Integrate biological and chemical suppression techniques with knowledge of behavior and dispersion to develop and test large area prevention and control strategies.
The following customers have expressed needs that will be met by this Research Unit: Armed Forces Pest Management Board (AFPMB), Department of Defense (DOD), Centers for Disease Control and Prevention, Environmental Protection Agency, USDA Animal, Plant and Health Inspection Service, World Health Organization, International Atomic Energy Agency, mosquito abatement districts, livestock producers, equine industry, pesticide control industry, food and restaurant industry, and the general public.
Year 2 (FY 2006) 1. Select top formulations and test in small outdoor plots. Scale up CuniNPV in vivo production system. Screen candidate cell lines with CuniNPV for susceptibility. 2. Generate antibodies for selected OB and ODV proteins. Localize and identify the selected ODV envelope proteins. 3. Screen mosquito tissue cultures with CPV and MIV to determine permissive lines. Continue sequencing of CPV genomes. 4. Collect and catalogue mosquito pathogenic viruses in Florida and Argentina 5. Finish multi-parasitism experiments. Begin release rate tests in large cages. Make inoculative releases on farms. 6. Assess fitness of Nosema-infected and healthy wasps. Treat wasps to eliminate Wolbachia. Describe microsporidia in parasitoids. 7. Test parasitoid responses to crude extracts; refine extracts by Liquid Chromatography (LC). 8. Scale-up insectary and High Through-put (HTP) bioassays; Strengthen and increase collaborations; Identify lead chemistry. 9. Perform additional evaluations, if necessary, especially with traps that give exceptionally good results. Conduct laboratory work on dosages and formulations of insecticides and fungi. Initiate large cage trials. Complete round 1 of stable fly management studies in Florida and Louisiana. 10. Complete examination of promising human-produced inhibitors that were identified in laboratory tests. Continue QSAR analysis of repellents, initiate synthesis of novel compounds. Formulate and optimize blends of human-produced inhibitors for maximum effectiveness in laboratory bioassays. Continue Gas Chromatograph/Mass Spectrophotometer (GC/MS) avian (chicken) odor analysis for candidate repellents.
Year 3 (FY 2007) 1. Conduct large scale field studies with top formulations in Florida; If susceptible cell lines are established, evaluate these candidate viruses for virulence and productivity. 2. Identify the proteins and their genes that are crucial to the CuniNPV infection process. 3. Publish CPV genome information. 4. Collect and catalogue mosquito pathogenic viruses in Florida. 5. Finish release rate tests in large cages; Assess establishment of released wasps. 6. Transfer technology to producers. Assess fitness of Wolbachia-infected and healthy wasps. 7. Determine composition of active fractions by GC/MS; bioassay compounds alone and in blends. 8. Expand bioassay species and methods; Selectively refresh and replace collaborations; Develop and formulate leading products for field trials. 9. Complete large cage trials. Refine trap design. Assess the impact of round 1 stable fly target studies and either make adjustments and perform round 2 or terminate trials. 10. Initiate GC/MS analysis of equine volatiles (from Sub-Objective 1.1.3 of Project # 6615-32000-040). Test promising inhibitor blend(s) in the field. Test novel repellent compounds for activity. Complete chemical analysis of chicken volatiles, initiate analysis of crow volatiles. Initiate bioassays of avian compounds for inhibition of mosquito host-seeking behavior.
Year 4 (FY 2008) 1. Evaluate top formulation in different locations (Connecticut and/or Wyoming); Scale-up in vitro production. 2. Examine host/viral protein interaction in the mosquito midgut; Test biological activity of viral proteins in mosquito bioassays. 3. Collect and catalogue mosquito pathogenic viruses in Florida. 4. Begin augmentative releases. 5. Transfer technology to insectaries. Assess fitness of Wolbachia-infected and healthy wasps. 6. Test candidate kairomones in the field. 7. Conduct a major review of the DOD's Deployed War Fighter Protection (DWFP) program and a reappraisal of known and potentially new insecticides. Review and revise bioassay procedures. Further refresh and replace collaborations. Scale-up field trials of leading products. 8. Initiate field tests of best traps. 9. Complete chemical analysis of equine hosts and continue with examination of avian (crow) hosts. Complete initial tests of human-based inhibitor blends in the field. Continue lab bioassay assessment of inhibitors and blends.
Year 5 (FY 2009) 1. Conduct cost-benefit analysis of CuniNPV for mosquito control. 2. Evaluate proteins with biological activity as novel control strategies. 3. Collect and catalogue mosquito pathogenic viruses in Florida. 4. Complete augmentative releases. 5. Conduct cross-mating studies for cytoplasmic incompatibility effects. 6. Sustain, refine and document progress all program activities from previous years, with critical scientific improvements. Transfer leading products to regulatory and commercial stages. 7. Continue field testing. 8. Continue with further refinement of inhibitor. Continue GC/MS analysis of avian volatiles, including grackles.
Refinement of an inhibitor field release system. Prototype devices that release inhibitors at a controlled rate have been developed by BioGuard through a Specific Cooperative Agreement with the MFRU. These devices were evaluated near Cairo, Egypt in August for their ability to reduce host-finding ability of Phlebotomus papatasi, a vector of leishmaniasis in the Middle East.
Evaluation of new repellents. During 2006, a Specific Cooperative Agreement with the Department of Chemistry at the University of Florida using Quantitative Structure Activity Relationship (QSAR) to discover and design effective novel repellents resulted in delivery of 33 piperdines and 12 carboxamides to the MFRU for testing. Once evaluated, these compounds will be used to further refine the QSAR model for repellent discovery.
First complete genome sequence for a Mosquito Iridescent Virus. A naturally-occurring mosquito iridescent virus (MIV) that kills mosquitoes has been isolated by MFRU scientists. This virus is designated as "Regular Mosquito Iridescent Virus" (RMIV) with strains isolated from a number of important mosquito species. In a collaborative effort, the complete genomic sequence of RMIV has been determined with an analysis of its relationship to other iridescent viruses as well as other pathogenic viruses. This DNA virus encodes 126 predicted genes, 27 which are shared homologues in all currently sequenced iridescent viruses, suggesting a genetic core for the Family Iridoviridae. This fundamental information contributes to our basic understanding of these mosquito pathogenic viruses at the molecular level and will enable a more complete evaluation and understanding how they might be exploited to control of mosquitoes that vector deadly diseases of man and animals.
New virus of black flies discovered. A naturally-occurring virus that infects black flies has been characterized by MFRU scientists. Biological and molecular studies have been conducted which have determined that this RNA virus has a 10-segmented genome and sequence analysis of the occlusion body protein has determined this species (SuCPV) is a new member of the Cypoviruses (SuCPV-20). This new virus from black flies expands the genetic diversity base required to investigate and understand basic mechanisms involved in infectivity and host range that will enhance capabilities to use these viruses for the control of black flies.
Visual traps for house flies. Four candidate insecticides were evaluated for use in visual targets for house flies. In field cages, targets treated with spinosad and imidacloprid provided nearly 100% control after 24 hours. Imidacloprid-treated targets remained effective after 1 month of natural weathering in the field.
Natural inhibition of Culex attraction. Culex mosquitoes are the principal vectors of West Nile virus and generally do not prefer to feed on humans. Humans are unique in producing high amounts of lactic acid on their skin compared to other animals such as birds and livestock. In laboratory studies, attraction of three species of Culex to potential hosts was decreased by the addition of lactic acid. This information provides the basis for development of naturally-occurring compounds from human skin that can preferentially deter Culex species from biting.
Evaluation of new toxicants for mosquitoes. Analyses of the structural activity of piperidines as toxicants against adult mosquitoes were conducted. Five additional chemicals were designed based on these findings and synthesized to confirm the structural activity relationship of the piperidines.
Air curtains used for disinsection. A system to prevent mosquitoes and flies from exiting commercial aircraft at terminals was further refined and is ready for transfer to the commercial sector. Final testing was completed and meetings were held with American Airlines officials to coordinate the protocol for deploying the system in the aircraft immediately after it reaches the terminal. Officials from the Ministry of Health (MOH) of Jamaica and the US Department of Transportation visited the MFRU to tour testing facilities, discuss future plans, and later participated in a demonstration of the system aboard an American Airlines Boeing 757 at Orlando International Airport. If the system was accepted by Jamaica and other countries currently requiring disinsection of aircraft, the use of pesticides on commercial aircraft would be reduced and perhaps eliminated. A pilot project with the Jamaican MOH and American Airlines is being discussed.
Hogsette, J.A. and D.A. Carlson. 2002. Insect trap. U.S. Patent application, Serial No. 10/145,787. Patent number 6,966,142, issued November 22, 2005.
Specific Cooperative Agreement with the Department of Chemistry, University of Florida. Objective is to design chemicals based on an evaluation of USDA legacy data that will be effective repellents against blood-sucking arthropods such as mosquitoes, sand flies, stable flies, black flies, tsetse flies, biting gnats, horse flies and deer flies, many of which transmit human and animal diseases.
Specific Cooperative Agreement with Connecticut Agricultural Experiment Station. Objective is to develop new methods and strategies to reduce the risk of human and domesticated animal infection with West Nile and other emerging infectious viruses carried by mosquitoes.
Specific Cooperative Agreement with BioGuard. Objective is to complete field development of devices to deliver MFRU-developed proprietary attractant-inhibitors.
Dr. Allan presented "Evaluation of human-associated odors on attraction of Culex" at the Annual Meeting of the Entomological Society of America, Ft. Lauderdale, FL. December 15-18 2005.
Dr. Becnel gave an invited presentation on "Biological control of mosquitoes" at a Florida Environmental Health Training Workshop, Tallahassee, FL, March 15, 2006.
Dr. Becnel gave an invited presentation on "Nosema bombycis and the silkworm industry" at the IX International Colloquium on Invertebrate Pathology and Microbial Control, 39th Annual Meeting of the Society for Invertebrate Pathology and VIII International Conference on Bacillus thuringiensis, Wuhan, China, August 2006.
Dr. Becnel gave an invited presentation on "Evolving life cycles of complex microsporidia in mosquitoes" at the 11th International Congress of Parasitology (ICOPA XI), Glasgow, Scotland U.K., August 6-11, 2006
Dr. Becnel's former Research Associate (Dr. Perera) presented "“Structural Proteins of Culex nigripalpus Nucleopolyhedrovirus (CuniNPV)" at the IX International Colloquium on Invertebrate Pathology and Microbial Control, 39th Annual Meeting of the Society for Invertebrate Pathology, and VIII International Conference on Bacillus thuringiensis, Wuhan, China, August 6-11, 2006.
Dr. Bernier presented "Mosquito attractant and repellent research progress," at the 3rd Arbovirus Surveillance and Mosquito Control Workshop, Anastasia Mosquito Control District (AMCD) Workshop, St. Augustine, FL, March 22-24, 2006.
Dr. Bernier presented "Chemistry strategies to identify host odors that attract ornithophilic mosquito species" at the 4th International Congress of Vector Ecology, Reno, NV, October 2-7, 2005.
Dr. Bernier presented "Development of mosquito attractants and repellents" at the Florida Department of Health Training Session, Tallahassee, FL, March 15, 2006.
Dr. Clark organized and co-moderated "Mosquito Vector Control and Biology in Latin America-A 16th Symposium" at the 72nd Annual Meeting of the American Mosquito Control Association, Detroit, MI, February 26 - March 2, 2006.
Dr. Clark made a presentation on "Dengue fever: a threat to the southeastern United States?" at a Florida Environmental Health Training Workshop, Tallahassee, FL, March 15, 2006.
Dr. Clark presented "Dengue fever: a threat to Florida?" at the 3rd Arbovirus Surveillance and Mosquito Control Workshop, Anastasia Mosquito Control District (AMCD) Workshop, St. Augustine, FL, March 22-24, 2006.
Dr. Clark presented "How to respond to a dengue epidemic: a global vision and experience from Puerto Rico" and "Surveillance and control of mosquitoes and prevention methods developed at the USDA" at the 3rd International Course on Integrated Prevention and Control of Dengue in Fortaleza, Ceara, Brazil, August 14-15, 2006.
Dr. Clark presented "West Nile virus activity in the Americas" at a workshop for vector control officials from Ceara state in Fortaleza, Ceara, Brazil, August 16, 2006.
Dr. Geden gave an invited keynote presentation, "Biological control of pests of animal husbandry" at the International Conference on Implementation of Biological Control in Temperate Regions, Flakkebjerg, Denmark, November 1, 2005.
Dr. Geden presented "Pathogens for control of beetles and flies in poultry systems" as an invited seminar speaker at the Department of IPM, Danish Institute of Agricultural Sciences, Lyngby, Denmark, November 4, 2005.
Dr. Geden gave an invited presentation, "Sampling methods to monitor pathogens associated with muscoid flies and fly dispersal" in a symposium on "Muscoid flies: pathogen dispersal and control" at the Annual Meeting of the Entomological Society of America, Fort Lauderdale, FL, December 2005.
Dr. Geden co-authored a paper presented by Dr. V. D. Blaeske entitled, "Sex and flies and birth control?" at the Annual Meeting of the Entomological Society of America, Fort Lauderdale, FL, December 2005.
Dr. Geden presented "Visual targets for management of house flies" at the Livestock Insects Workers Conference, Amarillo, TX, June 2006
Dr. Geden presented "Salivary gland hyperplasia virus of house flies; prevalence, transmission, and effects on fitness" at the meeting of the Society for Invertebrate Pathology, Wuhan, China, August 6-11, 2006.
Dr. Hogsette presented a seminar entitled "Nuisance fly management on Australian feedlots" in the First Friday Seminar Series, CMAVE, January 13, 2006.
Dr. Hogsette was invited to teach a day-long course entitled "Flies and other arthropods of public health importance" at the Florida Mosquito Control Association Dodd Short Courses, Ocala, FL, January 24, 2006.
Dr. Hogsette gave an invited lecture entitled "Biology and control of nuisance flies" at the Florida Environmental Health Association Training Session, Tallahassee, FL, March 15, 2006.
Dr. Hogsette was invited by the World Health Organization (WHO) to attend a planning meeting to discuss writing a chapter entitled "Flies," which he has co-authored with Dr. Jens Amendt, Frankfurt, Germany, and is included in the upcoming WHO book "Urban Pests and Health," London, England, March 23-24, 2006.
Dr. Hogsette was invited to attend the 2nd Annual Review of the ARS-DWFP Research Program and gave a presentation entitled "Flies and fly management," College Station, TX, March 27-28, 2006.
Dr. Hogsette attended several meetings of the Armed Forces Pest Management Board (AFPMB), Walter Reed Army Institute of Research, Silver Spring, MD. At the October, 2005 meeting, he was invited to revise, in collaboration with Dr. Graham White, Technical Guide 30, entitled "Flies." This was completed and presented to the AFPMB in March, 2006. At that meeting, the AFPMB requested that he evaluate commercial fly traps of potential interest to the military to determine efficacy in various climatic zones. At the July, 2006 meeting, he gave a presentation describing plans for the evaluation of fly traps near Cairo, Egypt in August, 2006.
Dr. Hogsette was invited to consult with members of the Florida Poultry Federation and attend their Annual Poultry Days Awards Banquet, Orlando, FL, April 29, 2006.
Dr. Hogsette, Dr. Carlson, Dr. Linthicum, and Dr. Clark met with Dr. Peter Knight and Ms. Sherine Huntley from the Ministry of Health of Jamaica and a representative of the US Department of Transportation in Gainesville for a demonstration and discussions about the air curtain system developed by Drs. Hogsette and Carlson for preventing flies and mosquitoes from escaping from commercial aircraft parked at terminals. They also viewed a demonstration of the system aboard an American Airlines Boeing 757 aircraft at the Orlando International Airport.
Dr. Hogsette attended the 50th Annual Livestock Insects Workers Conference and gave a presentation entitled "Evaluation of integrated techniques against nuisance fly populations on Australian feedlots" Amarillo, TX, June 25-28, 2006.
Dr. Kline co-organized and co-moderated the symposium "Vector-host interactions and trap development" and presented "Do commercial traps efficiently collect mosquitoes attracted to various hosts?" at the 4th International Congress of the Society of Vector Ecology, Reno, NV, October 2-6, 2005.
Dr. Kline's graduate student (Melissa Doyle) presented "Response of Aedes albopictus (Skuse) to control measures around the home" at the Annual Meeting of the Florida Mosquito Control Association, Duck Key, FL, November 15, 2005.
Dr. Kline was invited to teach a half-day course entitled "Trapping technology" at the Florida Mosquito Control Association Dodd Short Courses, Ocala, FL, January 26, 2006.
Dr. Kline presented "Attractant-baited traps for mosquito surveillance/management: present realities and future hopes" as an invited seminar for the Department of Entomology, North Carolina State University, Raleigh, NC, January 30, 2006.
Dr. Kline presented an invited lecture entitled, "Trapping technology for mosquito surveillance and control" at the Florida Environmental Health Association Training Session, Tallahassee, FL, March 15, 2006.
Dr. Kline presented "CO2: various sources and their usefulness for mosquito surveillance and control" at the 3rd Arbovirus Surveillance and Mosquito Control Workshop, St. Augustine, FL, March 22-24, 2006.
Dr. Kline presented "Impact of various modifications to the standard Nzi trap on collections of Tabanidae in the Lower Suwannee Wildlife Refuge," at the 50th Annual Livestock Insects Workers Conference, Amarillo, TX, June 25-28, 2006.
Dr. Linthicum gave an invited presentation, "Six years of West Nile virus expansion in the US: Are we prepared to contain a globalization of Rift Valley fever?" at the 77th Annual Meeting of the Florida Mosquito Control Association, November 13-16, 2005, Duck Key, FL. Dr. Linthicum presented, "The Center for Medical, Agricultural, and Veterinary Entomology: developing new mosquito surveillance and control products" at the 74th Annual Conference of the Mosquito and Vector Control Association of California, January 29-February 1, 2006, Reno, NV.
Dr. Linthicum gave an invited presentation, "Overview of research programs at the Center for Medical, Agricultural, and Veterinary Entomology (CMAVE), Agriculture Research Service, USDA, Gainesville, Florida" at the 2006 IAEA Training Course, May 10, 2006, Gainesville, Florida.
Dr. Pereira (Coordinator of the DWFP Program) gave a lecture entitled "The Deployed War-Fighter Protection (DWFP) Program: Role of the USDA-ARS Center for Medical, Agricultural and Veterinary Entomology (CMAVE)" at the spring meeting of the Florida Mosquito Control Association, St. Petersburg, FL, May 17-18, 2006.
Dr. Pereira presented a poster entitled "Role of the USDA-ARS in the Deployed War-Fighter Protection (DWFP) Program" at the 9th Annual Force Health Protection Conference, Albuquerque, NM, August 6-10, 2006.
McCauley, C. 2006. Entrapment. Horse Capital Digest, June 2006, pp. 36-37. Article describes mosquito research performed by Drs. Dan Kline and Jerry Hogsette (USDA/ARS/CMAVE/MFRU) and Dr. Saundra Ten Broeck and Aimee Holton (Department of Animal Sciences, University of Florida).
Reinert, J.F., Harbach, R.E. 2006. Descriptions of genus downsiomyia vargas (diptera: culicidae: aedini) and its type species do. nivea ludlow. Zootaxa. 1196:33-61.
Reinert, J.F. 2005. Redescription of the holotype of ochlerotatus eatoni edwards, 1916 (diptera: culicidae: aedini). European Mosquito Bulletin.
Broce, A.B., Hogsette Jr, J.A., Paisley, S. 2005. Winter feeding sites of hay in round bales as major developmental sites of stable flies, stomoxys calcitrans (diptera: muscidae), in pastures in spring and summer. Journal of Economic Entomology. 98(6):2307-2312.
Geden, C.J. 2005. Methods for monitoring outdoor populations of house flies, musca domestica l. (diptera: muscidae). Journal of Vector Ecology. 30(2):244-250.
Geden, C.J., Hogsette Jr, J.A. 2006. Suppression of house flies (diptera: muscidae) in florida poultry houses by sustained releases of muscidifurax raptorellus and spalangia cameroni (hymenoptera: pteromalidae). Environmental Entomology. 35(1):75-82.
Reinert, J.F., Harbach, R.E., Huong, V. 2006. Aedes (finlaya) alongi galliard & ngu (diptera: culicidae: aedini): description of the female, redescription of the fourth-instar larva, and reassignment to genus bothaella reinert. Zootaxa. 1135:49-55.
Kline, D.L. 2006. Coleman's MD-2500 Mosquito Trap: Industry's Best kept Secret? Abstracts of submitted papers, posters, and symposia presentations of 72nd AMCA Annual Meeting, February 26- March 2, 2006, Detroit, Michigan, #207, p42.
Geden, C.J., Moon, R.D., Butler, J.A. 2006. Host ranges of six solitary filth fly parasitoids (hymenoptera: pteromalidae, chalcididae) from Florida, Eurasia, Morocco, and Brazil. Environmental Entomology. 35(2):405-412.
Hogsette Jr, J.A., Carlson, D.A. 2005. Insect trap. U.S. Patent 6,966,142.
Green, T.B., Shapiro, A.M., White, S.E., Rao, S., Mertens, P.P., Carner, G., Becnel, J.J. 2006. Molecular and biological characterization of a cypovirus from the mosquito culex restuans. Journal of Invertebrate Pathology 91:27-34.
Geden, C.J. 2005. Biological control of arthropod pests in livestock production. Meeting Abstract. Flakkebjerg, Denmark. Nov. 1-3, 2005.
Becnel, J.J. 2006. Biological Control of Mosquitoes. Technical Bulletin of the Florida Mosquito Control Association. 7: 48-54.
Rongnoparut, P., Rodpradit, P., Kongsawadworakul, P., Sithiprasasna, R., Linthicum, K. 2006. Population genetic structure of Anopheles maculatus in Thailand. American Mosquito Control Association. 22:192-197.