Location: Arthropod-borne Animal Diseases Research
Project Number: 3020-32000-018-017-S
Project Type: Non-Assistance Cooperative Agreement
Start Date: Aug 1, 2023
End Date: Jul 31, 2026
Objective:
1. Determine relationship between antimicrobial resistance and pesticide resistance in house flies.
2. Identify and describe microbial communities associated with dipteran pests of livestock.
3. Investigate the sugar feeding ecology of Culicoides biting midges and potential for use as a control target.
Approach:
For Obj. 1, house flies will be reared in the presence of antimicrobials for several generations and susceptibility to pesticides such as permethrin will be assessed via bioassays. If antimicrobial exposure results in emergence of a resistance phenotype, flies will be collected for specific sensitivity assays (e.g. LD50, LC50) and genetic and/or gene expression analyses will be performed to identify the mechanism of resistance.
For Obj. 2, 16S and 18S gene profiling will be used to determine prokaryotic and eukaryotic microbial communities, respectively, that are associated with house flies and biting midges, including larval habitats. Community composition, species abundance and prevalence, species richness and other ecological analyses will be performed.
For Obj. 3, field collected biting midges will be tested for sugar (fructose) feeding by cold anthrone assays or similar methods. To determine the plant sources of fructose, DNA from additional field-collected midges will be amplified by PCR and visualized by gel electrophoresis, as has been developed for mosquitoes and will be adapted to biting midges. Amplicons will be sequenced and compared to reference databases for taxonomic identification of source plants. Behavioral assays will be used to determine attractive aspects of host plants that can be incorporated into the design of an attractive toxic sugar bait. Colony Culicoides sonorensis will be used in 2-, 3-, or 4-choice assays to investigate preferences for specific sugar-source attributes, such as color.
