1. Florida Medical Entomology Laboratory MOSQUITO CONTROL RESEARCH NEEDS: NEW PRIORITIES ASSESSING CURRENT AND PAST INVESTMENTS TO MAKE FURTHER PROGRESS Walter J. Tabachnick Florida Medical Entomology Laboratory University of Florida, IFAS February 12, 2008

2. Florida Medical Entomology Laboratory FCCMC Research Priorities -2007 TopicRank Score Non Pesticide Control166 Emerging Pathogens264 Pesticide - Non target Effects363 Disease Surveillance/Control/Risk Prediction462 Mosquito Ecology/Behavior561 Application Vegetative Barrier656 Domestic Mosquito Control/Storm Water755 Environmental Residue Monitoring854 Pesticide New Products954 Pesticide Efficacy/Resistance1053 Application Larvicide (e.g. canopy penetration)1152 Application Aerial Techniques1251 Public Education/Risk Communication1347 Mosquito Surveillance Trapping Systems1443 Attractants/Repellents1537

3. Florida Medical Entomology Laboratory Funded Research Projects 2007 Using water table to reduce emergency insecticide applications Isolating new mosquitocidal bacteria Increasing testing efficacy of mosquito pools Non pesticidal control pilot study Effects of droplet size, air blast, application angle for barrier treatment Spread, larval habitat, abundance, vector status of Culex coronator External morphology of Cx. nigripalpus female antennae Updating and reprinting the White Paper

4. Florida Medical Entomology Laboratory Funded Research Projects 2006 Statewide monitoring of mosquito response to organophosphates Evaluating automatic spray systems in backyards Improving mosquito control using permethrin aerial ULV while monitoring non-targets Isolating mosquitocidal bacteria Enhanced surveillance of arboviruses in Florida

5. Florida Medical Entomology Laboratory FCCMC Research Priorities -2007 TopicRank Score Non Pesticide Control166 Emerging Pathogens264 Pesticide - Non target Effects363 Disease Surveillance/Control/Risk Prediction462 Mosquito Ecology/Behavior561 Application Vegetative Barrier656 Domestic Mosquito Control/Storm Water755 Environmental Residue Monitoring854 Pesticide New Products954 Pesticide Efficacy/Resistance1053 Application Larvicide (e.g. canopy penetration)1152 Application Aerial Techniques1251 Public Education/Risk Communication1347 Mosquito Surveillance Trapping Systems1443 Attractants/Repellents1537 $32,252 $180,558 $68,300 $63,898 $23,837 $21,000 $115,737 $83,118 $119,362

6. Florida Medical Entomology Laboratory Data Gaps - Priorities Disease (Emerging Pathogens) Prediction/Control –WN, SLE, EEE, CHIK, DEN Products/Operations –New/Improved Products –Strategies to Reduce Pesticides MOSQUITO BIOLOGY

7. Florida Medical Entomology Laboratory Research Gaps/Projects - 2008 Improve Vector-Borne Disease Prediction and Mitigation in Florida For Arboviruses –Vector competence of Florida mosquitoes –Ecology – identify environmental and biological triggers for outbreaks, reduce pesticides. –Improved prediction/interventions –Improve mosquito keys Non Pesticide Products/Methods to Reduce Pesticides –Reduce pesticide reliance – i.e. mosquito biology, ecology, behavior, habitat information, targeting control for maximum effectiveness, improve mosquito keys –Improve effectiveness of Bti 10 fold using synergy with TMOF.

8. Florida Medical Entomology Laboratory Vector Competence of Florida Mosquitoes for Arboviruses Identify factors that control vector ability. Identify target mosquito populations for control at a time before they become dangerous vectors PRIORITY: ● THIS PROVIDES UNDERSTANDING RISK FROM EMERGING PATHOGENS, CURRENT PATHOGENS ● REDUCES UNNECESSARY CONTROL, TARGETS CONTROL ● REDUCES COSTS AND RELIANCE ON PESTICIDES

9. Florida Medical Entomology Laboratory WNV and SLEV in Florida Vectors We have found differential vector competence in Culex quinquefasciatus and Culex nigripalpus for WNV and SLEV attributed to the mosquito midgut. (Richards et al 2007, 2008)

10. Florida Medical Entomology Laboratory WNV and SLEV in Florida Vectors FMEL projects funded by NIH (Lord, Smartt, Richards, Anderson, Tabachnick) Identify critical midgut proteins Isolate and sequence proteins Construct primers to amplify controlling genes of each protein Screen DNA libraries to identify genes cDNA libraries for Cx. nigripalpus and Cx. p. quinquefasciatus for vector competence and pesticide resistance genes. PROJECT: OBTAIN GENE LIBRARIES

11. Florida Medical Entomology Laboratory Present MWTD recording sites (n = 589) PROJECT: Using KBDI data to evaluate arboviral transmission and the need for vector control Jon Day, Roxanne Connelly Priorities: Disease, Surveillance, Control Reducing pesticides

12. Florida Medical Entomology Laboratory Arboviral transmission risk for May 2005

13. Florida Medical Entomology Laboratory MWTD Data for the Pinellas County, FL WNV outbreak of 2005 Pinellas drought terminated WNV transmission

14. Florida Medical Entomology Laboratory KBDI data at 4 km resolution Project: Provide KBDI data for all of Florida Provide information to target mosquito and disease control efforts

15. Florida Medical Entomology Laboratory Dengue, chikungunya - Preparing Florida for Emerging Pathogens… Aedes aegyptiAedes albopictus

16. Florida Medical Entomology Laboratory PROJECT: Water-holding tree holes and leaf axils: assessing their importance in providing aquatic habitats for Aedes albopictus and Aedes aegypti Jorge R. Rey and George F. O’Meara PRIORITIES ● Disease Surveillance/Control/Risk Prediction ● Mosquito Ecology/Behavior ● Domestic Mosquito Control/Storm Water

17. Florida Medical Entomology Laboratory Information on habitat usage by container mosquitoes is needed. Aedes albopictus and Aedes aegypti also utilize natural containers such as, tree holes and leaf axils. The impact of natural containers in Florida on the production of Ae. albopictus and Ae. aegypti is not known.

18. Florida Medical Entomology Laboratory Neoregelia sp. Forty mosquito positive Neoregelia plants sampled No. plants with No. of larvae & pupae Wy. mitchellii 8 62 Wy. vanduzeei 26 590 Ae. albopictus 31 294 Ae. aegypti 23 122 Cx. quinquefasciatus 25 681

19. Florida Medical Entomology Laboratory ● Examine various plant species to assess their potential for providing aquatic habitats for Ae. albopictus and Ae. aegypti. ● Survey selected communities to determine the distribution and abundance of plants with a high propensity for frequently harboring immatures of Ae. albopictus and/or Ae. aegypti. ● Information derived from this study will assist mosquito control operations in targeting important mosquito production sites that might otherwise be overlooked. RESEARCH PROJECT

20. Florida Medical Entomology Laboratory Habitat characteristics at an intermediate scale influence A. aegypti and A. albopictus oviposition. FROM: Habitat Segregation of Mosquito Arbovirus Vectors in South Florida. Rey et al. 2006. J. Med. Entomol. 43: 1134-1141

21. Florida Medical Entomology Laboratory Long range determinants (field collections/ GIS) Land-use Geographic setting (surrounding habitats, distance from coast, etc.) Short range determinants (lab experiments) Preference of container background (vegetation, bricks, mixed, open). Container usage in Florida – (field surveys) Preferences Distribution Interaction with, short, medium and long range setting. Florida Medical Entomology Laboratory PROJECT: Determinants of Ae. aegypti and Ae. albopictus habitats - Jorge Rey

22. Florida Medical Entomology Laboratory Long range (kms) determinants of habitat selection. land use geography habitat type … Medium Range… Short range (meters) determinants of oviposition site selection. habitat composition access human activity … Container type preferences. type size shape, color, material abandoned/used ….

23. Florida Medical Entomology Laboratory Project: Identify factors for barrier treatments to be effective Cynthia Lord and Roxanne Connelly PRIORITIES: ● Mosquito Ecology/Behavior ● Application Vegetative Barrier ● Non pesticide – trapping barriers

24. Florida Medical Entomology Laboratory Barrier methods for control Increasing use to protect specific locations –Vegetation treatment with pesticides –Traps and attractants Goal: to block mosquitoes from reaching target Questions for success –What proportion of the population must be blocked? –How does timing or spacing affect control? –Other important factors: persistence, attraction distance, population characteristics? The tradeoffs between these and other factors in the success of a barrier are not well understood

25. Florida Medical Entomology Laboratory Proposed research Model: –Examine effect of key factors on level and duration of control in ideal landscape –Translate to more realistic landscape to compare to field studies –Identify factors or problems requiring further research Field Studies –Monitor existing control program –Compare with model studies This research will improve our understanding of the factors that affect the success of barrier control strategies

26. Florida Medical Entomology Laboratory 1. E-Z Key to the Genera of Florida Mosquitoes 2. Illustrated Taxonomic Keys to the Adult Female Mosquitoes of Florida Priorities: Mosquito Biology Surveillance Roxanne Connelly & Phil Lounibos Project: Modernize Florida Taxonomic Keys

27. Florida Medical Entomology Laboratory Project Deliverables for Mosquito Control Modernize and simplify taxonomic keys of Florida mosquitoes 3-D photography not line drawings Provide images in taxonomic keys –Card set of Common Mosquitoes of Florida was a huge success! Provide following publications: –“E-Z” Key to the Genera of Florida Mosquitoes - color laminated –Illustrated Taxonomic Keys to the Adult Mosquitoes of Florida (genera and species) –Illustrated keys to the 4 th instar larvae of Florida mosquitoes Other publications with high quality images

28. Florida Medical Entomology Laboratory Example of page from “Illustrated taxonomic keys to the adult female mosquitoes of Florida”

29. Florida Medical Entomology Laboratory Example of Images for New Taxonomic Keys

30. Florida Medical Entomology Laboratory Feeding Brewer’s Yeast Cells to Aedes aegypti Larvae for 4 Days [5mg dry cells/5 larvae/25 ml water]. PROJECT: Biopesticides for mosquito control Dov Borovsky Use genetically engineered yeast producing Ae. aegypti TMOF as a larvicide PRIORITIES: ● New products ● Non pesticide control

31. Florida Medical Entomology Laboratory Feeding Pichia pastoris_TMOF Cells to Aedes aegypti Larvae for 4 days [5mg dry cells (3.02  g TMOF)/5 larvae/25 ml water]. Effect of feeding genetically engineered yeast with TMOF to mosquito larvae

32. Florida Medical Entomology Laboratory Proposal: ● Test synergy with Bti ● Kills within 3 days ● Effective up to 3 rd instar ● Test in field plots ● No larval development ● Lethal ● Death within 10 days

33. Florida Medical Entomology Laboratory RESEARCH DATA GAPS Priorities and High Likelihood for Success and Impact Disease Surveillance/Control/Risk Prediction –Vector Competence – Gene Libraries –Environmental Triggers – KBDI for all of Florida Emerging Pathogens (CHIK, DEN, WNV, SLEV) –Vector Competence Mechanisms – Gene Libraries Mosquito Ecology/Behavior –Success of barrier/trapping treatments –Container mosquito habitats –Improved keys Non Pesticide Control/New Products –TMOF and synergy with Bti Domestic Mosquito Control –Container mosquito habitats