SELECTION OF ACTIVE INGREDIENTS FOR CONTROL OF MEDICALLY IMPORTANT DIPTERA

• Sponsoring Institution: State Agricultural Experiment Station
• Project Status: TERMINATED
• Funding Source: STATE
• Reporting Frequency: Annual
• Accession No.: 0209827
• Project No.: FLA-ENY-04616
• Project Start Date: Jun 1, 2006
• Project End Date: May 31, 2009

Project Director
Kaufman, P. E.

Recipient Organization
UNIVERSITY OF FLORIDA
G022 MCCARTY HALL
GAINESVILLE,FL 32611

Performing Department
ENTOMOLOGY & NEMATOLOGY

Non Technical Summary
There is a considerable lack of effective and safe chemicals for use against pestiferous flies, particularly those of medical importance. Development of new materials is a cumbersome and expensive process that requires years of research and high development costs. Due to overuse, misuse and lack of choices within which to rotate, mosquitoes and muscoid flies have become increasingly resistant to many currently available insecticides. Implementation of the Food Quality Protection Act has further reduced the numbers of acceptable materials. The end result is an even greater selection pressure and more rapid resistance development to remaining materials. 1. Identification and development of new insecticides for protection against blood feeding insects and filth flies will provide the military new and effective technologies to reduce exposure risk to military personnel. 2. The identification and development of new safe insecticides will present opportunities for solving problems of personal or spatial protection against vectors, especially, those not now controlled.

Animal Health Component

(N/A)

Research Effort Categories

Basic

20%

Applied

80%

Developmental

(N/A)

Classification

Knowledge Area (KA)	Subject of Investigation (SOI)	Field of Science (FOS)	Percent
721	6099	1130	100%

Knowledge Area
721 - Insects and Other Pests Affecting Humans;

Subject Of Investigation
6099 - People and communities, general/other;

Field Of Science
1130 - Entomology and acarology;

Keywords

mosquito

insecticide

sand fly

Goals / Objectives
Objective 1. Determine LD50 and LD90 values for the 20 identified semiochemicals for sand flies, Aedes mosquitoes and house flies. Objective 2. Which of the most promising insecticides, identified in Objective 1, are efficacious following prolonged exposure to environmental conditions? Objective 3. Which of the most promising insecticides identified in Objective 1 are efficacious against other blood-feeding dipterans, including Anopheles, Culex, Aedes Stomoxys and Culicoides species?

Project Methods
We will utilize L. shannoni (Dyar), a known New World vector of Leishmaniasis, as the test organism representing the Phlebotomine sand flies. The rearing method of Endris et al. (1982) will be utilized. Initially, all test subjects will be exposed to insecticides applied in serial dilution to a glass surface (30-ml Wheaton vial). For materials found inappropriate with the residue-on-glass approach, we will utilize topical application for house fly, and a treated chamber with a treated air stream using the olfactometer described earlier for the more volatile compounds. Utilization of the treated air stream will allow for determination of knock down suppression, repellency or attraction of these materials. Initial testing of each compound will be performed to generate a range of mortality of between 5 and 100 percent mortality. As these materials have not been previously studied with these species, we will experiment with duration of exposure and develop a standard testing protocol specific for each compound. Data will be analyzed using standard probit analysis (Finney 1971). Dose-mortality curves will be generated and LD50 and LD90 values determined. Objective 2. After identifying the most active compounds for each of the three test species outlined in Objective 1, we will evaluate the compounds through a two-phase process. Active compounds will be formulated into mixtures for appropriate testing. Phase 1: We anticipate evaluating insecticide-active materials on several types of cloth, including military uniform, bed netting and tent canvas cotton-based fabrics, polyester, and cotton-polyester blends. Treatment of painted and unpainted plywood surfaces, plastic/fiberglass and cinderblocks will also be examined. These studies will be comparative in design and include standard, untreated applications on glass. Test subjects will include the sand fly, Ae. aegypti and house fly previously described. Phase 2 testing will involve longer-term environmental exposures, such as heat exposure, ultraviolet exposure and time-since application variables. Environmental conditions will be generated both laboratory-equipment (heating ovens), as well as, natural exposure (solar exposure, rain, humidity) to conditions in Gainesville, Florida. Test subjects will include the sand fly, Ae. aegypti and house fly previously described. Application surfaces will include cloth, cinderblocks and plywood. Although we cannot control conditions, we will monitor pesticide response to the exposures that present themselves. Objective 3. We will evaluate, at a minimum, An. quadrimaculatus, Ae. albopictus, a Culex species. Stable fly adults will be reared using standard rearing procedures. Non-colonized species, such as Culicoides adults, will be field collected using CDC or Pickle Jar traps. We anticipate utilizing the appropriate testing procedures as outlined in Objective 1 (residue on glass, microapplicator and olfactometer test chamber). Compounds found most active in Objective 1 will be of primary interest.

Progress 06/01/06 to 05/31/09

Outputs
OUTPUTS: Studies identifying the toxicity of the semiochemicals have been completed for all flies identified in the project (house fly, stable fly, Aedes aegypti, Ae. albopictus Anopheles quadrimaculatus, Lutzomyia shannoni). The five most active compounds were formulated into usable products and were tested in laboratory conditions to refine their efficacy. Laboratory testing of these yielded two that were evaluated under field conditions with mosquitoes and house flies. Trapping for sand flies continued through 2008. Studies completed examined Lutzomyia shannoni attraction to a variety of olfactory and visual attractants and have concluded a 2-year study on seasonality. A small colony of L. shannoni was established and reared for several generations. This project supported a postdoctoral associate and a doctoral student who were mentored through this project. A patent application was submitted for 3 of the compounds. Information has been disseminated through oral presentations at regional and national entomology meetings and conferences with the USDA and military sponsors. Several publications has been published or are in press. An extension publication on sand flies has been published. PARTICIPANTS: Dr. Phillip Kaufman was the porject PI and oversaw the entire project. Drs. Jerry Butler and Pete Perkins assisted with the project as consultants and provided valuable information on the sand fly collection and rearing processes. Dr. Rajinder S. Mann was the postdoctoral associate on this project. Dr. Mann supervised the employees and the completion of the research objectives. Jimmy Pitzer was a doctoral student assigned to this project. Mr. Pitzer assisted in the bioassays component. Rebecca Hammel and Joe Peccora. These individuals were OPS employees assigned to insect collection, bioassay processing, insect rearing and other daily duties. The USDA-ARS-CMAVE was a partner organization. This unit provided several of the mosquito species tested. This project supported one Postdoctoral associate and one graduate student who received advanced training. It also supported numerous undergraduate students who learned about research and insect ecology through this project. TARGET AUDIENCES: The target audience for this project was the US military, with application of the resultant products destined for civilian use. Components of this project were used in the University of Florida's Medical and Veterinary Entomology courses (ENY4660, 4660L, 6665 and 6665L) taught by Dr. Kaufman. PROJECT MODIFICATIONS: Nothing significant to report during this reporting period.

Impacts
We have determined that light, whether reflected or directed, does not play a major role in sand fly attraction to our traps. Carbon dioxide remains a critical component of traps, but the addition of certain volatile mixtures results in significantly greater capture of Lutzomyia shannoni and a related species, Lutzomyia vexator, a reptile feeding sand fly. We have determined the seasonality of L. shannoni and Lutzomyia vexator, in north Florida. This knowledge will aid in determining the usefulness of trapping for these flies and in the epidemiology should any of the diseases that they are capable vectors of become established in Florida. The identification of effective formulations that we developed enhance our ability to find partners for their marketing. The identification of the three active compounds and their patenting (pending), will allow for the development of new, less toxic insecticides that may be used for the protection of military personnel and civilians.

Publications

• Obenauer, P.J., Kaufman, P.E., Allan, S.A., and Kline, D.L. 2009. Infusion-baited ovitraps to survey ovipositional height preferences of container-breeding mosquitoes in two Florida habitats. Journal of Medical Entomology. 46: 1507-1513.
• Bentley, M.T., Kaufman, P.E., Kline, D.L., and Hogsette, J.A. 2009. Response of adult mosquitoes to light emitting diodes placed in resting boxes and in the field. Journal of the American Mosquito Control Association 25: 285-291.
• Mann, R.S., Kaufman, P.E., and Butler, J.F. 2009. Lutzomyia spp. (Diptera: Psychodidae) response to olfactory attractant- and light-emitting diode-modified Mosquito Magnet X (MM-X) traps. Journal of Medical Entomology, 46: 1052-1061.
• Obenauer, P.J., Kaufman, P.E., Allan, S.A., and Kline, D.L. 2009. Host-seeking height preferences of Aedes albopictus (Diptera: Culicidae) in North-Central Florida suburban and sylvatic locales. Journal of Medical Entomology, 46: 900-908.
• Doyle, M.A., Kline, D.L., Allan, S.A., and Kaufman, P.E. 2009. Efficacy of residual bifenthrin applied to landscape vegetation against Aedes albopictus (Skuse) (Diptera: Culicidae). Journal of the American Mosquito Control Association, 25: 179-183.
• Mann, R. S., Kaufman, P. E. and Butler, J. F. 2007. A sand fly, Lutzomyia shannoni Dyar (Insecta: Diptera: Psychodidae: Phlebotominae). Gainesville, FL: IFAS Communications. EENY-421.
• Kaufman, P. E. (2007) Selection of Active Ingredients for Control of Medically-Important Diptera. Proceedings of the DWFP Annual Review 2007. Gainesville, FL, 03/08/2007.
• Kaufman, P. E. (2007) Techniques for Evaluating Insecticide Resistance in Filth Flies. Proceedings of the 2007 DoD Pest Management Workshop, , Jacksonville, FL. 02/14/2007.
• Kaufman, P.E., Nunez, S.J., Mann, R.S, Scharf, M.E. and Geden, C.J. 2009. Insecticide resistance in house flies collected from Florida dairies. Florida Entomological Society Annual Meeting, Ft. Meyers, FL.
• Mann, R.S., Kaufman, P.E., and Butler, J.F. 2009. Semiochemicals: An attractive alternative to conventional pesticides for mosquito and fly control program. Florida Entomological Society Annual Meeting, Ft. Meyers, FL.
• Obenauer, P.J., Kaufman, P.E., Allan, S.A., and Kline, D.L. 2009. Ovitrap efficacy using plant infusions to monitor vertical distribution of Aedes albopictus Skuse (Diptera: Culicidae) in North-Central Florida. American Mosquito Control Association, New Orleans, LA.
• Obenauer, P.J., Kaufman, P.E., Allan, S.A., and Kline, D.L. 2009. Infusion-baited ovitraps to monitor container-breeding mosquitoes in North-Central Florida suburban and sylvatic locales. Southeastern Branch Entomological Society of America Meeting, Montgomery, AL.
• Kaufman, P.E., Mann, R.S., and Butler, J.F. 2010. Evaluation of semiochemical toxicity to Aedes aegypti, Ae. albopictus and Anopheles quadrimaculatus (Diptera: Culicidae). Pest Management Science (in press).
• Mann, R.S. and Kaufman, P.E. 2010. Colonization of Lutzomyia shannoni (Diptera: Psychodidae) utilizing an artificial blood feeding technique. Journal of Vector Ecology. (in press).
• Mann, R.S. and Kaufman, P.E. 2010. The seasonal abundance of phlebotomine sand flies, Lutzomyia species in Florida. Journal of the American Mosquito Control Association (in press).
• Obenauer, P.J., Buss, L.J., and Kaufman, P.E. 2009. Utilizing Auto-montage technology for identifying field-collected container-inhabiting mosquito eggs. Journal of the American Mosquito Control Association 25: 517-520.

Progress 10/01/07 to 09/30/08

Outputs
OUTPUTS: Studies identifying the toxicity of the semiochemicals have been completed for Musca domestica and Aedes aegypti. Additionally, several of the compounds have been tested against Anopheles quadrimaculatus and the biting stable fly, Stomoxys calcitrans. The five most active compounds have been formulated into usable products and were tested in laboratory conditions to refine their efficacy. Laboratory testing of these yielded two that are currently under field evaluation with mosquitoes and house flies. Trapping for sand flies continued through 2008. Studies completed in 2008 have examined Lutzomyia shannoni attraction to a variety of olfactory attractants and have concluded a 2-year study on seasonality. Information has been disseminated through oral presentations at regional and national entomology meetings. A publication on the seasonality of the sand flies has been submitted. Two other publications are under development. An extension publication on sand flies has been published. PARTICIPANTS: Dr. Rajinder S. Mann, Postdoctoral Associate TARGET AUDIENCES: Nothing significant to report during this reporting period. PROJECT MODIFICATIONS: Not relevant to this project.

Impacts
We have determined the seasonality of Lutzomyia shannoni and a related species, Lutzomyia vexator, in north Florida. This knowledge will aid in determining the usefulness of trapping for these flies and in the epidemiology should any of the diseases that they are capable vectors of become established in Florida. The identification of effective formulations that we developed enhance our ability to find partners for their marketing.

Publications

• Obenauer, P.J., P.E. Kaufman and S.A. Allan. 2008. Do tigers prefer mixed forests: Attractiveness of North-Central Florida plant detritus for Aedes albopictus oviposition. Society for Vector Ecology Annual Conference, Ft. Collins, CO.
• Kaufman, P.E., J.F. Butler and C. Nelson. 2008. Evaluation of the Mosquito Sentinel 360 trap in Florida residential environments. Journal of the American Mosquito Control Association, (In Press).
• Obenauer, P.J., P.E. Kaufman and S.A. Allan. 2008. Oviposition response of Aedes albopictus (Skuse) (Diptera: Culicidae) to common North Central Florida plants. Southeastern Branch Entomological Society of America Meeting, Jacksonville, FL.
• Zettel, C. and P.E. Kaufman. 2008. Yellow Fever Mosquito, Aedes aegypti (Linnaeus) (Insecta: Diptera: Culicidae). Gainesville, FL: IFAS Communications. 7 pp. EENY-434.

Progress 10/01/06 to 09/30/07

Outputs
Studies identifying the toxicity of the semiochemicals have been completed for the house fly, Musca domestica and the yellow fever mosquito, Aedes aegypti. Additionally, several of the compounds have been tested against a potential malaria vector, Anopheles quadrimaculatus. The most active compounds are currently being formulated into useable products and are undergoing laboratory testing. Trapping for sand flies continues. Completed studies have examined Lutzomyia shannoni attraction to a variety of Light Emitting Diodes (LED's). We have also conducted studies examining attraction to reflected light and combinations of wavelengths. Studies examining L. shannoni capture when using various chemical attractants have been completed. A small colony of L. shannoni has been established. Information has been disseminated in several forms through this project. These include presentations at regional entomology meeting, conferences with the military and USDA annual reviews.

Impacts
We have determined that light, whether reflected or directed, does not play a major role in sand fly attraction to our traps. Carbon dioxide remains a critical component of traps, but the addition of certain volatile mixtures results in significantly greater capture of L. shannoni and a related species, Lutzomyia vexator, a reptile feeding sand fly.

Publications

• Kaufman, P. E. 2007. Selection of Active Ingredients for Control of Medically-Important Diptera. Proceedings of the DWFP Annual Review 2007. Gainesville, FL.
• Kaufman, P. E. 2007. Techniques for Evaluating Insecticide Resistance in Filth Flies. Proceedings of the 2007 DoD Pest Management Workshop, Jacksonville, FL.
• Mann, R. S., Kaufman, P. E. and Butler, J. F. 2007. A sand fly, Lutzomyia shannoni Dyar (Insecta: Diptera: Psychodidae: Phlebotominae). Gainesville, FL: IFAS Communications. EENY-421.