A High Throughput Screening Method to Identify Potential Pesticides

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A High Throughput Screening Method to Identify
Potential Pesticides for Mosquito Control
Mosquito and Fly Research Unit Center for
Medical, Agricultural, and Veterinary
Entomology USDA-ARS, 1600 SW 23rd Drive
Gainesville, FL 32608
Julia Pridgeon, Ph.D.
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High Throughput Screen Why?

• Very few pesticides are currently registered for
  mosquito control more are needed.
• Frequent use of currently available pesticides
  will result in increased insecticide resistance.
• Urgent need for novel pesticide discovery.
• Great need for a high throughput screening method.

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High throughput screen How?

• Day 1 Hatch Aedes aegypti eggs under vacuum
• Day 2 Sort 1st instar larvae to 24-well plates,
  5/well
• Add 1ml of water and food
• Add 10 ul of insecticide solution
  
• Day 3 Score bioassay data (dead/total)

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High throughput screen How?
Day 1
Aedes aegypti eggs
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HTS Fast and High Throughput

• Fast data return
• Days vs Weeks
• 2) Big capacity
• 1000s vs 100s per week
• 3 runs per week
• 480 per run
• total 1440 / week
• Compared to adult topical

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HTS Space Friendly
6 ? 8 48 cups
10 X 24 X 9 2160 wells
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HTS Cost Effective
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Correlation between larval and adult assays
against Aedes aegypti
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Correlation between larval and adult assays
against Aedes aegypti
- DNOC
(n18, plt0.001)
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Correlation between larval and adult assays
against Aedes aegypti
(n16, plt0.001)

• 3

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High throughput screen useful?
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High throughput screen valid?
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High throughput screen Summary

• A fast, inexpensive, and space-friendly HTS
  method was developed.
• Correlation was evaluated using 19 known
  pesticides in the HTS larval and adult (topical
  application) systems.
• Significant positive correlation was found
  between the HTS assay and the topical application
  method (p lt 0.001).
• Validated as a primary screen system 2,000
  chemicals have been evaluated and SAR trends
  were very clear.

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Acknowledgments
Mosquito and Fly Research Unit USDA-ARS-CMAVE
Deployed War-Fighter Protection Research Program
(DWFP)
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Any questions? Email julia.pridgeon_at_ars.usda.gov
Thank you for coming!