Source: UNIVERSITY OF ARKANSAS submitted to NRP
A NOVEL YEAST MICROENCAPSULATED FORMULATION OF FLURALANER FOR THE CONTROL OF LARVAL MOSQUITOES, BITING MIDGES, AND HOUSE FLIES
Sponsoring Institution
State Agricultural Experiment Station
Project Status
COMPLETE
Funding Source
Reporting Frequency
Annual
Accession No.
1026527
Grant No.
(N/A)
Cumulative Award Amt.
(N/A)
Proposal No.
(N/A)
Multistate No.
(N/A)
Project Start Date
Jul 1, 2021
Project End Date
Feb 25, 2025
Grant Year
(N/A)
Program Code
[(N/A)]- (N/A)
Recipient Organization
UNIVERSITY OF ARKANSAS
(N/A)
FAYETTEVILLE,AR 72703
Performing Department
Entomology
Non Technical Summary
Background: United States Military (USM) personnel, beneficiaries, and Department of Defense (DoD) civilians and contractors are affected by a wide range of biting and pestiferous insects both at home and abroad. When insect populations on a military installation are uncontrolled, force readiness suffers. The DoD maintains a list of approved insecticides for use in contingency operations. Despite the importance of controlling biting and nuisance fly populations on installations, only 5 products are available for mosquito larval control, and no products are available to target filth fly or biting midge larvae. Fluralaner, a recently approved ectoparasiticide, shows enormous promise as a broad-spectrum insecticide, and is highly active against a number of pestiferous fly species. We propose to assess the efficacy of a novel, microencapsulated fluralaner larvicide against multiple target species, and to determine whether fluralaner can provide enhanced vector control compared to commercial off-the-shelf (COTS) products currently in use by the DoD.Hypothesis: The proposed research will test the hypothesis that yeast microencapsulated fluralaner will provide significantly enhanced control (higher toxicity and/or lower dose) of larval mosquitoes, biting midges, and filth flies compared to currently available insecticides. Previous work has shown that larval mosquitoes and adult house flies are highly susceptible to fluralaner, with significantly lower LD50 values compared to the active ingredients in currently available insecticides, and that ingested formulations are likely to further enhance toxicity.Specific Aims: 1) conduct in vivo laboratory studies to assess product efficacy against medically important insect larvae, 2) conduct semi-field studies to assess product performance under natural conditions, and 3) pursue product commercialization.Study Design: In year 1, dose-response assays using larval mosquitoes, biting midges, and house flies will be conducted to determine product LD50/90 for each species. Microencapsulated fluralaner will also be compared to 3 COTS mosquito larvicides with national stock numbers (Vectobac, Abate, Altosid) using mosquito larval toxicity assays to determine whether the experimental product provides equivalent control as COTS products with less active ingredient. Due to the lack of technical data on biting midge susceptibility to fluralaner, modified WHO cone and contact toxicity assays will be used to determine the LD50 and KD50 dose for adult midges. In year 2, semi-field experiments will be conducted to determine appropriate reapplication intervals needed to maintain acceptable control, and to assess product performance under natural conditions. Because of key differences in the biology of the 3 study species, multiple application protocols will be tested in order to make best practices recommendations for achieving population control on installations.Relevance: The lack of effective larvicides for filth flies and biting midges represents a significant capability gap in the USM's vector control response. A more efficient mosquito larvicide, requiring less material to achieve acceptable control, helps the USM move towards the goal of becoming a more agile fighting force. Overall, novel compounds with new modes of action are critical for combating insecticide resistance in vector and pest populations globally.
Animal Health Component
50%
Research Effort Categories
Basic
0%
Applied
50%
Developmental
50%
Classification

Knowledge Area (KA)Subject of Investigation (SOI)Field of Science (FOS)Percent
72131101130100%
Knowledge Area
721 - Insects and Other Pests Affecting Humans;

Subject Of Investigation
3110 - Insects;

Field Of Science
1130 - Entomology and acarology;
Goals / Objectives
We will address three technical objectives related to the optimization and performance of the product. Achieving these objectives will allow us to compare microencapsulated fluralaner directly to available COTS products, as well as to develop precise application rates, intervals, and methods tailored to specific groups of pestiferous flies.Technical Objective 1. Determine the efficacy of yeast microencapsulated fluralaner against larval mosquitoes, biting midges, and house fliesSub-Objective 1.1. Optimize yeast microencapsulation of the active ingredientSub-Objective 1.2. Calculate the LD50 and LD90 dose for each species using dose-response assaysSub-Objective 1.3. Compare the levels of mosquito larval control achieved with microencapsulated fluralaner compared to currently available larvicidesTechnical Objective 2. Determine the topical and contact toxicity of fluralaner against adult biting midgesSub-Objective 2.1. Calculate the topical lethal dose (LD)50 and LD90 dose for C. sonorensis using modified WHO contact toxicity assaysSub-Objective 2.2. Calculate the contact knock-down dose (KD)/LD50 and KD/LD90 dose for C. sonorensis using modified WHO cone assaysTechnical Objective 3. Assess product performance under semi-field conditions and determine appropriate application intervals for target speciesSub-Objective 3.1. Determine how long a single application of microencapsulated fluralaner provides acceptable control (<90% emergence) of mosquito, midge, and house fly larvaeSub-Objective 3.2. Compare the efficacy of single vs. repeated applications of microencapsulated fluralaner to food waste for house fly control?
Project Methods
Insecticide formulationFluralaner will be microencapsulated into yeast using established methods. Product formulation will be validated using HLPC mass spectrometry. Dose-response experiments using larval Aedes albopictus, Culicoides sonorensis, and Musca domesticawill be conducted in the laboratory in order to calculate the LD50 for each species. 10-fold dilutions of the product will be applied to larval replicates, and mortality will be recorded at 24h.Comparison to COTS ProductsIn order to assess whether microencapsulated fluralaner is a more efficient insecticide than currently available commercial off-the-shelf products, the experimental product will be compared to the mosquito larvicides Abate, Vectobac, and Altosid at both their labeled rates and at equivalent concentrations. Products will be applied to replicates ofAe. albopictuslarval replicates, and mortality will be recorded at 24 h or 5d (Altosid experiments).Contact and topical toxicity of fluralaner on Culicoides biting midgesBecause fluralaner has never been assessed as a topical insecticide forCulicoides, we will conduct WHO contact and topical toxicity assays using adult femaleC. sonorensis. 10 fold dilutions of fluralaner formulated in acetone will be applied topically to midges at a rate of 20nL/midge and mortality will be recorded at 24h. Modified WHO cone assays will be used to assess contact toxicity. Filter papers will be treated with 10 fold dilutions of fluralaner, as above, and cones placed over filter papers. Midges will be introduced into the cones for 3min. Knock-down will be recorded after 60min and mortality will be recorded at 24h.Semi-fieldexposuresTo determine product efficacy under field conditions and to determine appropriate reapplication rates, semi-field exposure experiments will be conducted. Microencapsulated fluralaner will be applied to larval replicates ofAe. albopictus,C. sonorensis, andM. domesticaat the previously determined LD90 rate. Replicates will be held outdoors, exposed to natural conditions and sunlight. Mortality and emergence will be counted weekly, and new larvae will be added to replicates weekly, until emergence exceeds 90%. To simulate treatment of waste disposal sites for house fly population control, bucket traps will be baited with standardized food waste and treated with microencapsulated fluralaner. Traps will be open to colonization from wild flies. We will test the effect of single and repeated treatments of waste sites to determine the most effective application strategy.

Progress 07/01/21 to 02/25/25

Outputs
Target Audience:DoD Stakeholders • Armed Forces Pest Management Board Commercialization Partners • Rentokill, Clarke, MGK Vector Control Personnel Changes/Problems: Nothing Reported What opportunities for training and professional development has the project provided?One graduate student was trained under the project (Blythe Lawson). This student recieved numerous training and professional development opportunitiesincluding: Presentations at national conferences (Livestock Insect Workers Conference and Entomological Society of America). Training on HPLC techniques and analysis. Training on confocal microscopy. Grant writing training resulting in a successfully funded USDA-NIFA-AFRI predoctoral fellowship How have the results been disseminated to communities of interest?The results of this research were disseminated to communities of interest via three peer reviewed publications as well as multiple conference presentations at the Livestock Insect Workers Conference and Entomological Society of America annual meeting. Results were also disseminated to the Armed Forces Pest Management Board (funding agency) through invited presentations at the Deployed Warfigther Protection Program Research Symposium and an organized DWFP program symposium at ESA (2024). A final technical report was submitted to the DWFP program officer in July 2024. What do you plan to do during the next reporting period to accomplish the goals? Nothing Reported

Impacts
What was accomplished under these goals? Technical Objective 1. Complete. All Sub-Objectives were acheived. Fluralaner was successfully microencapsulated and LC doses were calculated for Musca domestica, Aedes albopictus, and Culicoides sonorensis. YME fluralaner was compared to two commercial off the shelf larvicides for mosquito control (Vectobac and Altosid), and was found to perform equivalently, or to outperform, those products for Ae. albopictus control. A provisional patent application and PCT application was filed for the technology. All claims for the PCT application were deemed novel with inventive step and industrial application The results for TO1 were published as Lawson and McDermott 2024 (Acta Tropica-Invited paper) and Briggs et al. 2024 (Arthropod Management Tests) Technical Objective 2. Complete. All Sub-Objectives were achieved. Topical and contact LC50/LC90 were calculated for adult C. sonorensis. Additionally, oral applications of fluralaner (in a blood meal) were evaluated and found to have the lowest LC values. The results for TO2 were published as Lawson and McDermott 2023 (Parasites & Vectors) Technical Objective 3. Complete. All Sub-Objectives were achieved. Semi-field experiments were used to determine the efficacy of YME fluralaner under natural conditions. The product provided excellent control for Ae. albopictus out to 5 weeks and for C. sonorensis for at least 8 weeks. Poor control of filth flies was observed, but trials sufferred from inconsistency due to weather conditions. The results for TO3 were published asLawson and McDermott 2024 (Acta Tropica-Invited paper)

Publications

  • Type: Peer Reviewed Journal Articles Status: Published Year Published: 2024 Citation: Briggs, C., B.E. Lawson, and E.G. McDermott. 2024. Comparison of yeast microencapsulated fluralaner to commercially available mosquito larval control products, 2023. Arthropod Management Tests. 49: tsae105.
  • Type: Peer Reviewed Journal Articles Status: Published Year Published: 2024 Citation: Lawson, B.E., and E.G. McDermott. 2024. Successful yeast microencapsulation of fluralaner and its potential as a larvicide for vector control. Acta Tropica. 259: 107358.
  • Type: Conference Papers and Presentations Status: Published Year Published: 2024 Citation: E. G. McDermott, and B. Lawson. Developing yeast microencapsulated fluralaner as a larvicide for vector control Entomological Society of America National Meeting, Phoenix, AZ (2024)
  • Type: Conference Papers and Presentations Status: Published Year Published: 2023 Citation: E. G. McDermott and B. Lawson. Development and evaluation of microencapsulated fluralaner as an insecticide Deployed Warfighter Protection Program Research Symposium, Silver Spring, MD (2023)


Progress 10/01/22 to 09/30/23

Outputs
Target Audience: DoD Stakeholders Armed Forces Pest Management Board Commercialization Partners Nissan Chemical, Rentokill Vector Control Personnel Changes/Problems: Nothing Reported What opportunities for training and professional development has the project provided?One graduate student was trained under the project (Blythe Lawson). This student recieved numerous training and professional development opportunities in the last reporting period, including: Presentations at two national conferences (Livestock Insect Workers Conference and Entomological Society of America). Training on HPLC techniques and analysis. Training on confocal microscopy. Co-authorship on three published or pending manuscripts. How have the results been disseminated to communities of interest?The results of this project were disseminated in the following formats: Peer-reviewed publication Lawson and McDermott 2023. Parasites & Vectors. 16: 281. Conference presentations Deployed Warfighter Protection Program Research Symposium. Silver Spring, MD. February 2023. Livestock Insect Workers Conference. Fredericksburg, TX. June 2023. Entomological Society of America Annual Meeting. National Harbor, MD. November 2023. What do you plan to do during the next reporting period to accomplish the goals?A no-cost extension was granted to extend the period of performance for the project to April 2024. During this time, we plan to engage with Phoreus Biotechnologies, an outside company with which we have an MTA, to explore additional procedures to increase encapsulation yield of our product.

Impacts
What was accomplished under these goals? Technical Objective 1. Complete. All Sub-Objectives were acheived. Fluralaner was successfully microencapsulated and LC doses were calculated for Musca domestica, Aedes albopictus, and Culicoides sonorensis. YME fluralaner was compared to two commercial off the shelf larvicides for mosquito control (Vectobac and Altosid), and was found to perform equivalently, or to outperform, those products forAe. albopictuscontrol. A provisional patent application was filed for the technology. A publication on these results is pending. Technical Objective 2. Complete. All Sub-Objectives were achieved. Topical and contact LC50/LC90 were calculated for adultC. sonorensis. Additionally, oral applications of fluralaner (in a blood meal) were evaluated and found to have the lowest LC values. These results were published in the journal, Parasites & Vectors. Technical Objective 3. Complete. All Sub-Objectives were achieved. Semi-field experiments were used to determine the efficacy of YME fluralaner under natural conditions. The product provided excellent control forAe. albopictusout to 5 weeks and forC. sonorensisfor at least 8 weeks. Poor control of filth flies was observed, but trials sufferred from inconsistency due to weather conditions. A publication on these results is pending.

Publications

  • Type: Conference Papers and Presentations Status: Accepted Year Published: 2023 Citation: McDermott, E., (Presenting Author) Lawson, B., Deployed Warfighter Protection Program Research Symposium, "Development and evaluation of microencapsulated fluralaner as an insecticide," Silver Spring, MD. (March 2023).
  • Type: Conference Papers and Presentations Status: Accepted Year Published: 2023 Citation: Lawson, B. (Presenting Author), McDermott, E., Entomological Society of America Annual Meeting, "Successful Microencapsulation of Fluralaner and its Potential as a Larvicide for Vector Control," National Harbor, MD. (November 2023).
  • Type: Journal Articles Status: Published Year Published: 2023 Citation: Lawson, B. E., and E. G. McDermott. 2023. Topical, contact, and oral susceptibility of adult Culicoides biting midges (Diptera: Ceratopogonidae) to fluralaner. Parasites & Vectors. 16: 281.


Progress 10/01/21 to 09/30/22

Outputs
Target Audience: Nothing Reported Changes/Problems: Nothing Reported What opportunities for training and professional development has the project provided? Nothing Reported How have the results been disseminated to communities of interest? Nothing Reported What do you plan to do during the next reporting period to accomplish the goals? Nothing Reported

Impacts
What was accomplished under these goals? Nothing reported in FY22

Publications


    Progress 07/01/21 to 09/30/21

    Outputs
    Target Audience: Nothing Reported Changes/Problems:Thus far, the only change to report is that funding was recieved several months after the proposed project start date. This has delayed project initiation somehwhat in that we have not yet begun dose-response assays as anticipated. We were able to establish insect colony lines prior to the arrival of funds, and this has reduced the impact of the funding delay on experimental progress. What opportunities for training and professional development has the project provided?This work is primarily being conducted by a PhD student. This student has received training on insect colony maintainence as well as HPLC metholody, both new skills she has developed through working on this study. How have the results been disseminated to communities of interest? Nothing Reported What do you plan to do during the next reporting period to accomplish the goals?During the next reporting period, we will validate the insecticide formulation and complete dose-response experiments with larvalM. domestica, Ae. albopictus, andC. sonorensis. The remaining supplies for HPLC have been ordered and are expected to arrive this month. We will also complete contact toxicity and cone assay experiments using adultC. sonorensis. Protocols for these experiments are currently under development and review, and assays can begin as soon as this process is complete. These experiments are not contingent on completion of the microencapsulation validation, and so can be run concurrently. During the next reporting period we will also begin ordering supplies to conduct the sub-objective 1.3 experiments, as well as developing protocols for these experiments.

    Impacts
    What was accomplished under these goals? Funds for this project were delayed by several months, pushing back the planned project initiation date from May 2021 to October 2021. Technical Objective 1. Sub-Objective 1.1.We are currently working with Drs. Jack Lay and Jennifer Gidden of the UA Department of Chemistry to develop and validate HPLC protocols for insecticide formulation validation. We have also developed protocols for the microencapsulation procedure. Sub-Objective 1.2. New colony lines of M. domestica and Ae. albopictus have been established in our laboratory for use in these studies. An established colony of C. sonorensiswas acquired from the USDA under a MTA and is currently being reared in our laboratory as well. Preliminary experiments have been conducted using sham-encapsulated yeast and M. domestica in order to confirm that the insecticide solvent does not have insecticial effects, and that the encapsulation procedure does not negatively affect insect survival. Protocols for conducing the dose-responseexperiments are under development. Sub-Objective 1.3.Work on this sub-objective is dependent on completion of sub-objectives 1.1 and 1.2 and has not yet begun. Technical Objective 2. Sub-Objective 2.1.Protocols for conducing contact toxicity assays with adultC. sonorensisare currently under development. Sub-Objective 2.2.Protocols for conducing WHO cone assays with adultC. sonorensisare currently under development. Technical Objective 3. Work on Technical Objective 3 is scheduled to begin during year two of the study.

    Publications