Progress 06/01/08 to 05/31/11
Outputs
Target Audience: Target audiences are researchers in academia interested in agricultural production, industry involved in agricultural production, government agencies involved in regulating agricultural production, private groups interested in agricultural production, private and government groups interested in environmental quality, growers, the military interested in the control of vector arthropods, global human health organizations of any type, and local, state and federal government involved in arthropod control to prevent the transmission of diseases. The work conducted also has potential in the use and regulation of transgenic crops designed for insect control and therefore would be of interest to any groups where their mission is the production or regulation of these types of technologies. Changes/Problems: The work was conducted on one moth species to take advantage of opportunities to expand the scope of the research to plant bugs, stink bugs, kudzu bugs and two species of mosquitoes. In respect to the latter, this gave us the opportunity to obtain matching funds from the US Navy in the form of a three year student stipend to conduct the mosquito related work. What opportunities for training and professional development has the project provided? The project is part of 1 Ph.D. dissertation and all of a second Ph.D. dissertation in Entomology at NC State University. Several additional students and postdocs both in the US and abroad have participated on this proposal and are listed as co-authors on the papers published. The research has provided training in insect rearing, insect biology and physiology, insecticide toxicology, insect insecticide resistance managment, vector biology, technology transfer and commercialization. How have the results been disseminated to communities of interest? The research has been presented at several national and international scientific meetings in the form of oral talks and posters and has resulted in several published papers and two Ph.D. dissertations. We have also worked with extension agents in the state and the US navy to deploy the technology (projects ongoing). Futhermore, the work has resulted in several new research proposals at different levels of consideration. We also have provided work shops to growers on how to use our assays for the management of insect resistance to pesticides. What do you plan to do during the next reporting period to accomplish the goals? This project is continuing in a variety of new work initiatives as follows: (i) research proposals have been submitted with the US Navy to further develop plastic ware and field test the mosquito assay in Lima, Peru for use by the military; (ii) a research proposal is pending for developing the technology for monitoring insect susceptibility to dsRNA and siRNA; (iii) a SBIR Phase I grant is pending with the DOD to develop the technology into a mosquito control system; (iv) two proposals are pending with the USDA to further develop the kits (a) for monitoring for kudzu bug resistance and (b) to study kudzu bug microbiomes; and (v) the work has led to the development of a high throughput feeding bioassay system for evaluating mosquito attractants and repellents (funded by two grants from DOD).
Impacts
What was accomplished under these goals?
The work was limited to one moth species to expand the project to other sucking pests of agricultural importance outside of moths (i.e., plant bugs, stink bugs and kudzu bugs) and to sucking pests of importance outside of agriculture, i.e., mosquitoes (two species). The latter was supported in part by funding for an additional Ph.D. student on the project from the US Navy with the goal of using the technology on moths for deployed soldier protection from diseases carried by mosquitoes. We were successful in developing a novel assay plate architecture for both the moth and mosquito applications. We were able to use our existing plate architecture for plant bugs, stink bugs and kudzu bugs. We also developed diagnostic doses for the field evaluation of multiple chemical insecticides for all of the insect species listed under this section of accomplishments. In respect to the moth and mosquito research, studies were also conducted to validate the assay for field conditions, especially in the case where moths and mosquitoes of unknown ages are collected.
Publications
- Type:
Conference Papers and Presentations
Status:
Published
Year Published:
2009
Citation:
van Kretschmar, J.B., L.C. Magalhaes, J. Zhu, R.M. Roe and A.C. Cohen. 2009. Feasibility of a novel feeding disruption test (FDT) bioassay kit for rapid resistance detection of sucking pests of cotton. Proceedings, 2009 Beltwide Cotton Conferences, San Antonio, Texas, Jan. 5-8, 2009. National Cotton Council, Memphis, TN. 882-892
- Type:
Conference Papers and Presentations
Status:
Published
Year Published:
2011
Citation:
van Kretschmar, J.B., A. Dhammi, D. Reisig and R.M. Roe. 2011. Challenges for rearing of the stink bug: successes, failures, and RNAi screening. Proceedings, 2011 Beltwide Cotton Conferences, Atlanta, Georgia, Jan. 4-7, 2011. National Cotton Council, Memphis, TN. 1169-1175.
- Type:
Conference Papers and Presentations
Status:
Published
Year Published:
2011
Citation:
van Kretschmar, J.B., J.R. Bradley, A.R. Cabrera and R.M. Roe. 2011. Lepidopteran adult feeding disruption test (FDT) to detect insecticide resistance. Proceedings, 2011 Beltwide Cotton Conferences, Atlanta, Georgia, Jan. 4-7, 2011. National Cotton Council, Memphis, TN. 1010-1015.
- Type:
Journal Articles
Status:
Published
Year Published:
2013
Citation:
van Kretschmar, B., A.R. Cabrera, J.R. Bradley and R.M. Roe. 2013. Novel adult feeding disruption test (FDT) to detect insecticide resistance of lepidopteran pests in cotton. Pest. Manag. Sci. 69: 652-660.
- Type:
Journal Articles
Status:
Published
Year Published:
2012
Citation:
Stell, F.M., R.M. Roe, C. Arellano, L. Kennedy, H. Thornton, K. Saavedra-Rodriguez, D.M. Wesson, W.C. Black and C.S. Apperson. 2012. Proof of concept for a novel insecticide bioassay based on sugar feeding by adult Aedes aegypti (Stegomyia aegypti). Med. Vet. Entomol. doi: 10.1111/j.1365-2915.2012.01048.x.
- Type:
Journal Articles
Status:
Accepted
Year Published:
2013
Citation:
Stell, F.M., R.M. Roe,C. Arellano and C.S. Apperson. 2013. Innovative sugar-insecticide feeding bioassay for adult female Anopheles gambiae (Diptera: Culicidae). J. Med. Entomol. In press.
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Progress 06/01/10 to 05/31/11
Outputs
OUTPUTS: The overall goal of the project was to use feeding disruption as an end point for measuring insect resistance to pesticides, using the adult stage of moths. There are a number of advantages of this approach and potential for the work to be translational into several new areas of science. The research included the determination of proof of concept in the laboratory of using feeding disruption of moths on dyed nectar as an assay end point for intoxication from insecticides and a measure of insecticide susceptibility to permethrin in wild type and resistant insects, the conduct of laboratory experiments to show the technique will work in the field, the design of a prototype resistance monitoring kit, and the application of the concepts to other insect systems. The work resulted in a product with the potential for commercialization and is available for licensing; and the development of unexpected additional intellectual property unrelated directly to the original goals of the work (also available for licensing). The project also supported the Ph.D. work of at least four graduate students (three have completed their Ph.D.s in Entomology and one is on their final year of the Ph.D.), two visiting International scientists, and two US postdocs. The work has also resulted in additional funding from Cotton Inc., for translational work which is currently funding a student from the US military and in other projects which are at various stages of commercial development and grant writing. Our research has been presented at many scientific meetings, published in the proceedings for scientific meetings, is included in one Ph.D. dissertation already completed with another in progress, presented to companies interested in the intellectual property and in various stages of publication for scientific journals and book chapters. PARTICIPANTS: The project supported the Ph.D. work of four graduate students (three have completed their Ph.D.s in Entomology and one is on their final year of the Ph.D.), two visiting International scientists, and two US postdocs. The work has also resulted in additional funding from Cotton Inc., for translational work which is currently funding a student from the US military, and in other projects which are at various stages of commercial development and grant writing. The work has produced a number of inventions that are at various stages of commercialization. TARGET AUDIENCES: The target audiences include academic scientists working in basic and applied research in insecticide use and management and in USDA and private industry with similar interests. The work also includes new inventions for commercialization and of interest to private industry. PROJECT MODIFICATIONS: No major changes needed in the science to complete the goals of the project. However, unexpected was the use of the technology for other insect applications in and outside of agriculture, e.g., for the control of medically important pests, and in the process of conducting the work, the discovery of new inventions in genetic pest management with high potential for commercialization; both should result in follow up funding, research and commercial development. Also unexpected, the vast majority of the funding for this project was returned to the granting agency because of an error in understanding the terms of the contract by NC State University.
Impacts
Proof of concept was demonstrated both in the laboratory and for use for field collected insects that feeding disruption for moths on nectar could be used to measure larval and adult susceptibility to a variety of insecticides and to measure insect resistance development to pesticides. The work also resulted in the development of a prototype device or kit that can easily be used in the field or the laboratory for the routine monitoring and management of insect resistance to pesticides. The invention has the advantage of allowing for resistance detection to most mechanisms of resistance at the same time. The assay is rapid with an easy to read end point. The technique and device is designed for easy mass production, economy of space to facilitate transport and reduce cost, and ease of operation so minimal training is required to conduct the assay. The work has also been translational and resulted in the development of similar technology for other insect groups including other sucking pests, e.g., plant bugs, stink bugs, etc., and insects of importance to human health, e.g., mosquitoes. The work has also been translational in that it has resulted in a new invention in the area of insect genetic pest management, which is available for commercialization and at the time of this report being evaluated by two companies. The work has also led to the development of applications in biofuel production, new approaches to insect control (including the use of RNAi), and risk assessment associated with RNAi-based pest control.
Publications
- van Kretschmar, J. B. (2010) Novel Insecticide Resistance-Monitoring Bioassays for Lepidopteran and Hemipteran Cotton Pests and 454 Pyrosequencing to Identify Potential Gene Targets for RNAi Silencing in Hemipteran Cotton Pests. Ph.D. Dissertation. North Carolina State University, Raleigh, NC.
- van Kretschmar, J. B., L. C. Magalhaes, J. Zhu, A. C. Cohen and R. M. Roe. 2009. Feasibility of a novel feeding disruption test (FDT) bioassay kit for rapid resistance detection of sucking pests of cotton, pp. 882-892. In Proceedings Beltwide Cotton Conferences. National Cotton Council, Memphis, TN.
- van Kretschmar, J. B., J. R. Bradley, A. R. Cabrera and R. M. Roe (2008) Novel adult assay to detect insecticide resistance of Lepidopteran pests in cotton, pp. 1329-1336. In Proceedings Beltwide Cotton Conferences. National Cotton Council, Memphis, TN.
- van Kretschmar, J. B., J. R. Bradley, A. R. Cabrera and R. M. Roe (2011) Lepidopteran adult feeding disruption test (FDT) to detect insecticide resistance, pp. 1010-1015. In Proceedings Beltwide Cotton Conferences. National Cotton Council, Memphis, TN.
- van Kretschmar, J. B., J. R. Bradley, C. F. Stumpf and R. M. Roe (2007) Feasibility of adult feeding disruption tests (FDT) for monitoring Lepidopteran resistance to chemical insecticides, pp. 136-140. In Proceedings Beltwide Cotton Conferences. National Cotton Council, Memphis, TN.
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Progress 06/01/09 to 05/31/10
Outputs
OUTPUTS: OUTPUTS: (1) Developed a prototype device to measure the disruption of nectar feeding in moths to evaluate the use of this approach for monitoring insecticide resistance; (2) Evaluated the use of dyes as a marker for measuring nectar feeding in moths and that could be used to determine feeding disruption; (3) Evaluated the effect of nectar feeding followed by starvation on additional nectar feeding; (4) Examined the dose response of insecticides in nectar on moth feeding and mortality; and (5) Evaluated the use of feeding disruption as determined by the production of blue feces for the diagnosis of insecticide susceptibility. The work conducted is being presented at scientific meetings, published in scientific journals, used to support intellectual property disclosures and support advanced degrees in Entomology at NC State University. PARTICIPANTS: The research is the Ph.D. work in Entomology of Mr. Jaap B. van Kretschmar. This has provided training in bioassay development, insect resistance to pesticides, field and laboratory research related to the project, insect rearing, and applications of the work into other fields like transcriptomics and sucking pests. The expansion work has led to new intellectual property and the submission of a US patent and associated training. The PI, R. M. Roe, has been responsible for the idea of the project and its implementation including publications completed and in process, intellectual property disclosures, patents submitted, chair of Mr. van Kretschmar's graduate committee, expansion of the work into other fields as described in this report and presentations at scientific meetings. Many other graduate students, postdocs and other professors listed on the publications in this reporting also contributed to this project. The work also has been in cooperation with Cotton Inc. of Cary, NC (USA). TARGET AUDIENCES: The target audiences for the work include Ph.D. scientists, extension agents, students training for advanced degrees in Entomology and related fields, and private industry, especially companies developing insecticides and small companies interested in technology development. The work is being presented at local, national and international scientific meetings, presented in courses taught at NC State University, published in the scientific literature, submitted as NC State University intellectual property, submitted for patenting in the US and abroad, and marketed by NC State University for commercial development. PROJECT MODIFICATIONS: No major changes in the project were made. As a result of the research, new project areas have been developed as described in the progress report.
Impacts
(1) We were successful in developing a prototype device that can be used in proof of concept research and from which a final device for resistance assay can be modeled; (2) We developed a blue marker dye that can be added to nectar and that will not affect nectar feeding by moths and which is not absorbed from the nectar by the moth after feeding; (3) For partial proof of concept, it was necessary to show that if moths are collected from the field that have already fed on a nectar source, that starvation in the lab will initiate nectar feeding again. This was demonstrated successfully in the lab, i.e., that nectar fed moths when starved will feed again; (4) We showed that a dose response was possible for moths feeding on nectar containing in separate experiments, two different insecticide chemistries with different modes of action and with the end point being reductions in nectar consumption and mortality; and (5) We showed that feeding reductions or feeding disruption due to intoxication with insecticide in the nectar could be determined by reductions in the production of blue feces deposited on the assay device. In summary, we now have a prototype architecture for a feeding disruption assay for detecting insecticide resistance in moths and proof of concept that the assay method can detect insecticide susceptibility on laboratory reared insects. We also have traveled for training to rear a new insecticide resistance strain of the tobacco budworm for the final proof of concept for the technology under development and have developed assay architecture for production of a self running, field assay kit. In addition, the technology has been extended to proof of concept for larval and adult plant bugs and stink bugs and this work is being expanded. Furthermore, the assay has been extended to adult mosquitoes for the management of resistance to treated bed nets and area wide control using insecticides both abroad and in the US; this has resulted in the submission of a proposal to the military (pending)and the successful funding of a graduate student position in the lab by the US Navy to work on this application. Furthermore, we have additional proof of concept that the assay method will diagnose resistance using a laboratory pyrethroid resistant strain of the tobacco budworm. The work is part of a Ph.D. thesis with an anticipated completion of the thesis for graduation in December 2010.
Publications
- van Kretschmar, J. B., L. C. Magalhael, J. Zhu, R. M. Roe and A. C. Cohen. 2009. Feasibility of a novel feeding disruption test (FDT) bioassay kit for rapid resistance detection of sucking pests of cotton. Proceedings, 2009 Beltwide Cotton Conferences, San Antonio, TX, January 5-8, 2009. National Cotton Council, Memphis, TN. pp 882-892.
- Roe, R. M., K. V. Donohue, L. C. Magalhaes and J. van Kretschmar. 2009. First 454 transcriptome to the plant bug digestive system: new leads for next generation transgenic cotton to control sucking pests. Proceedings, 2009 Beltwide Cotton Conferences, San Antonio, TX, January 5-8, 2009. National Cotton Council, Memphis, TN. pp 1152-1158.
- Roe, R. M., H. P. Young, T. Iwasa, C. F. Wyss, C. F. Stumpf, T. C. Sparks, G. B. Watson, J. J. Sheets and G. D. Thompson. 2009. Mechanism of resistance to spinosyn in the tobacco budworm, Heliothis virescens. Pestic. Biochem. Physiol. 96: 8-13.
- van Kretschmar, J.B., K. V. Donohue, A. R. Cabrera, L. C. Magalhaes, C. E. Sorenson, J. S. Bacheler, S. M. S. Khalil and R. M. Roe. 2010. Transcriptomics by massive parallel, pyrosequencing of the green stink bug: functional gene ontology and new targets for control. Proceedings, 2010 Beltwide Cotton Conferences, New Orleans, LA, January 4-7, 2010. National Cotton Council, Memphis, TN. pp. 1195-1202.
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Progress 06/01/08 to 05/31/09
Outputs
OUTPUTS: (1) Developed a prototype device to measure the disruption of nectar feeding in moths to evaluate the use of this approach for monitoring insecticide resistance; (2) Evaluated the use of dyes as a marker for measuring nectar feeding in moths and that could be used to determine feeding disruption; (3) Evaluated the effect of nectar feeding followed by starvation on additional nectar feeding; (4) Examined the dose response of insecticides in nectar on moth feeding and mortality; and (5) Evaluated the use of feeding disruption as determined by the production of blue feces for the diagnosis of insecticide susceptibility. The work conducted is being presented at scientific meetings, published in scientific journals, used to support intellectual property disclosures and support advanced degrees in Entomology at NC State University. PARTICIPANTS: The project is providing research training and serving as part of the dissertation research for two Ph.D. students in Entomology at NC State University. The project also has resulted in preliminary studies of the use of feeding disuption on adult moths for resistance monitoring to the same for sucking pests, for example, plant bugs and stink bugs. In addition, the work has led to preliminary studies of using nectar feeding to detect resistance in adult mosquitoes and a proposal to the Gates foundation on new adult assay methods for insecticide resistance management related to the control of malaria and other vector-borne diseases. The project on moths is also a collaboration with applied cotton field, research and extension entomologists at NC State University and with Cotton Inc. TARGET AUDIENCES: The target audiences for the technology being development are research entomologists at the state, Federal and industrial levels and for those responsible for insecticide use, insecticide resistance management and integrated pest management in agriculture. PROJECT MODIFICATIONS: No major changes in the project are needed.
Impacts
(1) We were successful in developing a prototype device that can be used in proof of concept research and from which a final device for resistance assay can be modeled; (2) We developed a blue marker dye that can be added to nectar and that will not affect nectar feeding by moths and which is not absorbed from the nectar by the moth after feeding; (3) For partial proof of concept, it was necessary to show that if moths are collected from the field that have already fed on a nectar source, that starvation in the lab will initiate nectar feeding again. This was demonstrated successfully in the lab, i.e., that nectar fed moths when starved will feed again; (4) We showed that a dose response was possible for moths feeding on nectar containing in separate experiments, two different insecticide chemistries with different modes of action and with the end point being reductions in nectar consumption and mortality; and (5) We showed that feeding reductions or feeding disruption due to intoxication with insecticide in the nectar could be determined by reductions in the production of blue feces deposited on the assay device. In summary, we now have a prototype architecture for a feeding disuption assay for detecting insecticide resistance in moths and proof of concept that the assay method can detect insecticide susceptibility on laboratory reared insects.
Publications
- van Kretschmar, J. B., J. R. Bradley, A. R. Cabrera and R. M. Roe. 2008. Novel adult assay to detect insecticide resistance of lepidopteran pests in cotton. Proceedings of the 2008 Beltwide Cotton Conferences, Nashville, Tennessee. National Cotton Council, Memphis, TN.
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