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      Novel Mutations in the Voltage-Gated Sodium Channel of Pyrethroid-Resistant Varroa destructor Populations from the Southeastern USA

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          Abstract

          The parasitic mite Varroa destructor has a significant worldwide impact on bee colony health. In the absence of control measures, parasitized colonies invariably collapse within 3 years. The synthetic pyrethroids tau-fluvalinate and flumethrin have proven very effective at managing this mite within apiaries, but intensive control programs based mainly on one active ingredient have led to many reports of pyrethroid resistance. In Europe, a modification of leucine to valine at position 925 (L925V) of the V. destructor voltage-gated sodium channel was correlated with resistance, the mutation being found at high frequency exclusively in hives with a recent history of pyrethroid treatment. Here, we identify two novel mutations, L925M and L925I, in tau-fluvalinate resistant V. destructor collected at seven sites across Florida and Georgia in the Southeastern region of the USA. Using a multiplexed TaqMan® allelic discrimination assay, these mutations were found to be present in 98% of the mites surviving tau-fluvalinate treatment. The mutations were also found in 45% of the non-treated mites, suggesting a high potential for resistance evolution if selection pressure is applied. The results from a more extensive monitoring programme, using the Taqman® assay described here, would clearly help beekeepers with their decision making as to when to include or exclude pyrethroid control products and thereby facilitate more effective mite management programmes.

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          Insect Pollinated Crops, Insect Pollinators and US Agriculture: Trend Analysis of Aggregate Data for the Period 1992–2009

          In the US, the cultivated area (hectares) and production (tonnes) of crops that require or benefit from insect pollination (directly dependent crops: apples, almonds, blueberries, cucurbits, etc.) increased from 1992, the first year in this study, through 1999 and continued near those levels through 2009; aggregate yield (tonnes/hectare) remained unchanged. The value of directly dependent crops attributed to all insect pollination (2009 USD) decreased from $14.29 billion in 1996, the first year for value data in this study, to $10.69 billion in 2001, but increased thereafter, reaching $15.12 billion by 2009. The values attributed to honey bees and non-Apis pollinators followed similar patterns, reaching $11.68 billion and $3.44 billion, respectively, by 2009. The cultivated area of crops grown from seeds resulting from insect pollination (indirectly dependent crops: legume hays, carrots, onions, etc.) was stable from 1992 through 1999, but has since declined. Production of those crops also declined, albeit not as rapidly as the decline in cultivated area; this asymmetry was due to increases in aggregate yield. The value of indirectly dependent crops attributed to insect pollination declined from $15.45 billion in 1996 to $12.00 billion in 2004, but has since trended upward. The value of indirectly dependent crops attributed to honey bees and non-Apis pollinators, exclusive of alfalfa leafcutter bees, has declined since 1996 to $5.39 billion and $1.15 billion, respectively in 2009. The value of alfalfa hay attributed to alfalfa leafcutter bees ranged between $4.99 and $7.04 billion. Trend analysis demonstrates that US producers have a continued and significant need for insect pollinators and that a diminution in managed or wild pollinator populations could seriously threaten the continued production of insect pollinated crops and crops grown from seeds resulting from insect pollination.
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            Honey bee viruses.

            Viruses are significant threats to the health and well-being of the honey bee, Apis mellifera. To alleviate the threats posed by these invasive organisms, a better understanding of bee viral infections will be of crucial importance in developing effective and environmentally benign disease control strategies. Although knowledge of honey bee viruses has been accumulated considerably in the past three decades, a comprehensive review to compile the various aspects of bee viruses at the molecular level has not been reported. This chapter summarizes recent progress in the understanding of the morphology, genome organization, transmission, epidemiology, and pathogenesis of honey bee viruses as well as their interactions with their honey bee hosts. The future prospects of research of honey bee viruses are also discussed in detail. The chapter has been designed to provide researchers in the field with updated information about honey bee viruses and to serve as a starting point for future research.
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              DDT, pyrethrins, pyrethroids and insect sodium channels.

              The long term use of many insecticides is continually threatened by the ability of insects to evolve resistance mechanisms that render the chemicals ineffective. Such resistance poses a serious threat to insect pest control both in the UK and worldwide. Resistance may result from either an increase in the ability of the insect to detoxify the insecticide or by changes in the target protein with which the insecticide interacts. DDT, the pyrethrins and the synthetic pyrethroids (the latter currently accounting for around 17% of the world insecticide market), act on the voltage-gated sodium channel proteins found in insect nerve cell membranes. The correct functioning of these channels is essential for normal transmission of nerve impulses and this process is disrupted by binding of the insecticides, leading to paralysis and eventual death. Some insect pest populations have evolved modifications of the sodium channel protein which prevent the binding of the insecticide and result in the insect developing resistance. Here we review some of the work (done at Rothamsted Research and elsewhere) that has led to the identification of specific residues on the sodium channel that may constitute the DDT and pyrethroid binding sites.
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                Author and article information

                Contributors
                Role: Editor
                Journal
                PLoS One
                PLoS ONE
                plos
                plosone
                PLoS ONE
                Public Library of Science (San Francisco, CA USA )
                1932-6203
                18 May 2016
                2016
                : 11
                : 5
                : e0155332
                Affiliations
                [1 ]Department of Biological Chemistry and Crop Protection, Rothamsted Research, Harpenden, Hertfordshire, United Kingdom
                [2 ]ERI BIOTECMED, Department of Genetics, Universitat de València, Valencia, Spain
                [3 ]Department of Entomology and Nematology, University of Florida, Gainesville, Florida, United States of America
                [4 ]Bayer Animal Health GmbH, Leverkusen, Germany
                University of Crete, GREECE
                Author notes

                Competing Interests: Klemens Krieger is an employee of Bayer Animal Health GmbH, Leverkusen, Germany ( https://animalhealth.bayer.com/ah/) and received support in the form of salary. Joel González-Cabrera was supported by a grant from Bayer Animal Health GmbH but is not an employee of Bayer. There are no patents, products in development or marketed products to declare. This does not alter the authors' adherence to PLOS ONE policies on sharing data and materials.

                Conceived and designed the experiments: JGC MSW DS JDE. Performed the experiments: JGC SRV DS. Analyzed the data: JGC MSW. Contributed reagents/materials/analysis tools: JGC SRV DS MSW. Wrote the paper: JGC TGED LMF DS JDE KK MSW.

                [¤a]

                Current address: Instituto de Agroquímica y Tecnología de Alimentos-CSIC, Valencia, Spain

                [¤b]

                Current address: Bayer Division of Crop Science, Pollinator Safety, Research Triangle Park, North Carolina, United States of America

                Article
                PONE-D-16-09601
                10.1371/journal.pone.0155332
                4871586
                27191597
                458ff8be-c8df-4146-8cd2-07edf26cf98b
                © 2016 González-Cabrera et al

                This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.

                History
                : 6 March 2016
                : 27 April 2016
                Page count
                Figures: 3, Tables: 1, Pages: 10
                Funding
                Funded by: Marie Curie Intra European Fellowship
                Award ID: PIEF-GA-2011-298831
                Award Recipient :
                Funded by: Bayer Animal Health GmbH
                Award Recipient :
                Funded by: funder-id http://dx.doi.org/10.13039/501100007136, Secretaría de Estado de Investigación, Desarrollo e Innovación;
                Award ID: RYC-2013-13834
                Award Recipient :
                Funded by: funder-id http://dx.doi.org/10.13039/501100000268, Biotechnology and Biological Sciences Research Council;
                Award Recipient :
                Funded by: funder-id http://dx.doi.org/10.13039/501100000268, Biotechnology and Biological Sciences Research Council;
                Award Recipient :
                Funded by: funder-id http://dx.doi.org/10.13039/501100000268, Biotechnology and Biological Sciences Research Council;
                Award Recipient :
                Funded by: funder-id http://dx.doi.org/10.13039/501100000268, Biotechnology and Biological Sciences Research Council;
                Award Recipient :
                Funded by: funder-id http://dx.doi.org/10.13039/501100000268, Biotechnology and Biological Sciences Research Council;
                Award Recipient :
                Funded by: Florida Department of Agriculture (US)
                Award Recipient :
                Funded by: Florida Department of Agriculture (US)
                Award Recipient :
                Funded by: Bayer Animal Heath GmbH
                Award Recipient :
                Joel González-Cabrera was supported by a Marie Curie Intra European Fellowship within the 7th European Community Framework ( http://ec.europa.eu/research/mariecurieactions/), by Bayer Animal Health GmbH, Leverkusen, Germany ( https://animalhealth.bayer.com/ah/), and by the Spanish Ministry of Economy and Competitiveness, Ramón y Cajal Program ( http://www.mineco.gob.es/). Rothamsted Research is supported by the Biotechnology and Biological Sciences Research Council of the United Kingdom ( http://www.bbsrc.ac.uk/). University of Florida contributions to this research were supported by the Florida Department of Agriculture and Consumer Services, Division of Plant Industry, Apiary Inspection Section, through the guidance of the Florida Honey Bee Technical Council ( http://www.freshfromflorida.com/). Klemens Krieger is an employee at Bayer Animal Health GmbH, Leverkusen, Germany ( https://animalhealth.bayer.com/ah/). These funders provided support in the form of salaries for authors JGC, SRV, TGED, LMF, DS, KK and MSW but did not have any additional role in the study design, data collection and analysis, decision to publish, or preparation of the manuscript. The specific roles of these authors are articulated in the Author Contributions section.
                Categories
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                Organisms
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