1. Update on Duet™ and insecticide-treated surfaces Sandra A. Allan Center for Medical, Agricultural and Veterinary Entomology ARS/ USDA Gainesville FL DoD Pest Management Workshop Jacksonville NAS February 12 2010

2. Update 1. Effect of ULV droplets of DUET™ on mosquito (Culex quinquefasciatus) responses 2. Behavioral effects of ULV droplets of DUET™ on the sand fly (Lutzomyia shannoni) 3. Effect of different insecticides on surfaces on landing by mosquitoes and sand flies

3. Hypothesis Prallethrin contained in Duet enhances flight activity and results in greater mortality Objective Quantify mosquito excitation and mortality caused by Duet and its components applied as an ultra-low-volume aerosol Military Relevance Duet may be a ULV formulation useful for military applications Duet™ New product 1. Effect of ULV droplets of DUET™ on Culex quinquefasciatus responses

4. Treatments ComponentsABCDE Prallethrin-1% - Sumithrin5%-- - Piperonyl butoxide5%- - Inert Ingredients+++++ Formulations obtained from Clarke Mosquito Control Sprays delivered at a sublethal rate so that behavior can be observed sublethal

5. Side view of wind tunnel air flow Fan Intake Output Pesticide filters Filters Laminizer Reducer Door Pesticide cloud delivery Door Laminizer * screened cage with mosquito Filters 4 ft o Video recording  Individual mosquitoes placed in screened cage in wind tunnel downwind of spray inlet * pre-spray post-spray spray 5 min 2.5 min Video recordings were 12.5 min long:

6. Video Analysis using behavioral analysis software (Observer) Behaviors of individual insects are coded with start and stop times by different keys and data exported to Excel for analysis Event panel Assay cage in wind tunnel Event time Behavior

7. Video Analysis using motion analysis software (Motus) Manual tracking of individual insect produces a flight track and x-and y- coordinates that are exported to Excel for further analysis Assay cage in wind tunnel Cursor on top of insect Resulting flight track

8. ULV Droplet analysis  Mosquitoes treated in the wind tunnel were dissected and body parts placed on slides  Droplets were measured and counted 50 microns

9. Conclusions Behavior: (pre-spray compared with spray and post-spray)  Prallethrin produced increased flight activity (excitation) during spray  Sumithrin produced increased flight activity post-spray  Controls: no differences in behavior were observed Mortality: Increased mortality appears to be linked to increased activity levels during spray, especially when exposed to treatments with prallethrin

10. Increased flight results in increased droplet contact Droplets Pre-spraySpray Post-spray

11. Objective Quantify sand fly excitation and mortality caused by Duet™ and its components applied as an ULV spray Methods Similar to study with mosquitoes except:  Used field-collected Lutzomyia shannoni  Smaller assay cages  Double screening to contain sand flies 2. Behavioral effects of ULV droplets of DUET™ on the sand fly (Lutzomyia shannoni)

12. Detection of ULV droplets of Duet on body parts Droplets most detected on wings of sand flies Sand fly wing 50 microns

13. Droplet size distribution – Comparison between sand flies and mosquitoes More small droplets detected on sand flies than on mosquitoes Sand fly wing 10 microns

14. Mortality of sand flies after treatment with Duet formulations in wind tunnel Treatments containing prallethrin Treatments containing prallethrin had higher mortality at 24 hr

15. Sumithrin, PBO + inerts Prallethrin, PBO + inerts Prallethrin + inerts Duet (sumithrin, prallethrin, PBO + inerts) Control (inerts) Flight tracks of sand flies (Lutzomyia shannoni) exposed to ULV spray of Duet in wind tunnel During spray More movement by sand flies exposed to prallethrin

16. Conclusions  Similar to mosquitoes, sand fly activity and mortality is enhanced in the presence of prallethrin

17. If given a choice, will mosquitoes land on surfaces treated with residual pesticides? Are there differences between mosquitoes and sand flies? Objective To compare landing of mosquitoes and sand flies on surfaces treated with different residual pesticides Military Relevance To identify the most effective available residual pesticides for control of mosquitoes and sand flies 3. Effect of different insecticides on surfaces on landing by mosquitoes and sand flies

18. Active ingredientFormulation AI (%) BifenthrinTalstar One 7.9 CyfluthrinTempo Ultra (SC) 11.8 DeltamethrinSuspend SC 4.75 PermethrinDragnet 36.8 Lambda-cyhalothrinTriazicide 0.002% Methods Selected residual insecticides approved for application on vegetation and other surfaces for mosquito control Applied to surfaces at maximum label rate

19. Methods SpeciesAedes albopictus (lab-reared) Lutzomyia shannoni (field-collected) Phlebotomus papatasi (lab-reared) ApproachEach test cage with: Filter paper Wax myrtle leaves Treated surface Control surface

20. Methods – Mosquito videotaping Assay cage Camera

21. Methods – Sand fly videotaping Assay cage Camera

22. Preliminary Conclusions  Mosquitoes were more sensitive than sand flies to pesticide-treated substrates  Repellent responses were stronger to treated filter paper than to treated leaves which likely reflects a higher dose on the filter paper  Landing responses were least affected by lambdacyhalothrin and most decreased by permethrin

23. Acknowledgements CMAVE Gary Clark Miriam Cooperband (USDA/APHIS) Erin Vrzal Fran EllisonWilliam Jany, Clarke Mosquito Control Larry Pitts Eric Paulsen Lee Cohnstaedt This study was supported in part by funds from the Deployed War-Fighter Protection Research Program from the U.S. Department of Defense through the Armed Forces Pest Management Board to the Agricultural Research Service of the U.S. Department of Agriculture.