Source: OHIO STATE UNIVERSITY submitted to
SUSTAINABLE SOLUTIONS TO PROBLEMS AFFECTING BEE HEALTH
Sponsoring Institution
National Institute of Food and Agriculture
Project Status
TERMINATED
Funding Source
HATCH
Reporting Frequency
Annual
Accession No.
1004928
Grant No.
(N/A)
Project No.
OHO01355-MRF
Proposal No.
(N/A)
Multistate No.
NC-_old1173
Program Code
(N/A)
Project Start Date
Nov 28, 2014
Project End Date
Sep 30, 2019
Grant Year
(N/A)
Project Director
Johnson, RE, M..
Recipient Organization
OHIO STATE UNIVERSITY
1680 MADISON AVENUE
WOOSTER,OH 44691
Performing Department
Entomology
Non Technical Summary
Insect pollinators provide essential pollination services to growers of U.S. fruits, vegetables, nuts and seeds. Honey bees are the premier managed pollinator and account for $11.7 billion of the $15 billion of agricultural output attributable to insect-mediated pollination (Calderone, 2012). To satisfy the demand for pollination about 2 million of the 2.6 million managed honey bee colonies in the U.S. are rented and placed in nearly 100 different crops each year.Efficient delivery of managed pollination services is threatened by the poor state of U.S. honey bees. Since the mid-2000s beekeepers have consistently witnessed the loss of 30-35% of colonies over winter (Spleen et al., 2013). While beekeepers can often make up for these losses through intensive management of surviving colonies, current management tools are costly and may not be sufficient to indefinitely sustain the honey bee colony numbers or colony strength needed for pollination.Wild pollinators also contribute substantially to agricultural pollination in many crops (Garibaldi et al., 2013). Unfortunately, the long-term health and abundance wild pollinators is under threat as well.The causes of honey bee and pollinator declines in the U.S. are varied, complex, and defy a simplistic explanation, as multiple stressors are almost certainly involved. Significant progress in identifying contributing factors to bee declines has been made by many current members of the NC1173 multi-state project through a $4.1M, 4-year USDA CAP project that was funded in 2008 to study the causes of Colony Collapse Disorder (CCD) and other factors affecting bee populations. Current members are also part of the $5M CAP through the USDA Global Food Security program to establish the Bee Informed Partnership, an extension-only effort to collect and disseminate information about the health of the managed bee population.Many of the findings from these large collaborative projects were presented and synthesized at the Stakeholders Conference on Honey Bee Health convened by the USDA and the U.S. Environmental Protection Agency in October 2012. The summary of this conference provides a roadmap for future research to be addressed by members of the NC1173 multi-state project:1. Parasites and pathogens - The Varroa mite and the viruses it helps to transmit remain a top concern for beekeepers. The gut parasite Nosema has been implicated some honey bee colony losses and other species of Nosema affect managed and wild bumble bees. A range of other bacteria, fungi, and animals negatively affect bee health. Improved understanding of the interaction between bees and their parasites and pathogens will yield better management and control strategies.2. Breeding and genetic diversity - Breeding resistance to parasites and pathogens in bees is a viable means to mitigate colony losses, but stock improvement through breeding is in no way complete and success will only be achieved through constant incremental improvement. Additionally, the genetic diversity and mating success of honey bees also plays a role in colony success.3. Forage availability and nutritional stress - The nutritional requirements of honey bees and other pollinators are not met by the floral landscape in some parts of the U.S. Research is needed into land- and farm-management practices associated with high levels of colony and pollinator success.4. Pesticides and environmental contaminants - Insecticides designed to kill insects may harm pollinating insects as well. Other pesticides and environmental contaminants also have the potential to affect bees. Additionally, drugs used to control pests and pathogens may have unintended side-effects. Work is needed to determine the effects of pesticide exposure on colony health, honey production and delivery of pollination services.The consensus is that these multiple stressors, working in concert, are together responsible for the honey bee and pollinator health issues manifest in the U.S. While advances are currently being made in all four of these key research areas, a real solution to honey bee and pollinator health will only come by taking them all together a task that is too big and too complex to be managed by any researcher working independently. As such, the collaborative work fostered by the NC1173 multi-state research project is critical to building a holistic understanding of honey bee and pollinator health.As such, there is a clear need defined by the stakeholders to mitigate the continued decline of honey bees and other insect pollinators. The consequences of inaction are a further destabilized food-production system, decreased yields and quality of fruits and vegetables, and potentially higher produce prices. The technical feasibility of the proposed working group is greatly facilitated by the existing practice of adjoining the American Bee Research Conference (ABRC) the annual professional meeting of the American Association of Professional Apiculturists (AAPA) with one of the three national apiculture associations in the US in alternating years: the American Beekeeping Federation (ABF), the American Honey Producers of America (AHPA), and the Apiary Inspectors of America (AIA). This tradition of interfacing with the clientele and other professional groups concerning beekeeping is ideally suited to collaboration, interaction, and discussion of current problems that face the industry. Thus there is a clear advantage of fostering this multi-state effort, because there is great similarity in the threats to American beekeeping across all regions. The impacts from these ongoing interactions have been significant (see above), and therefore a continuation of the NC1173 working group will advance these successes going forward. We should note that a CRIS search was conducted for the expiring NC1173, so there is no overlap with any ongoing projects.
Animal Health Component
0%
Research Effort Categories
Basic
40%
Applied
40%
Developmental
20%
Classification

Knowledge Area (KA)Subject of Investigation (SOI)Field of Science (FOS)Percent
3143120113050%
3143010113050%
Knowledge Area
314 - Toxic Chemicals, Poisonous Plants, Naturally Occurring Toxins, and Other Hazards Affecting Animals;

Subject Of Investigation
3120 - Spiders, mites, ticks, and other arthropods; 3010 - Honey bees;

Field Of Science
1130 - Entomology and acarology;
Goals / Objectives
To determine how land management practices affect pollinator nutrition and how nutrition affects honey bee colony productivity and success To assess the effects of exposure to pesticides and other xenobiotics on the survival, health and productivity of honey bee colonies and pollinator abundance and diversity To determine the effects of interactions among various factors affecting pollinator and honey bee colony health To develop and recommend "best practices" for beekeepers, growers, land managers and homeowners to promote honey bee and pollinator health
Project Methods
Objective 3 -- To determine how land management practices affect pollinator nutrition and how nutrition affects honey bee colony productivity and success1) We will study how land use and the diversity of foraging resources affect the growth, development, and health of honey bee colonies by experimentally placing hives into landscapes that vary in floral resource quality and diversity, and subsequently measuring variables related to hive health. For example, we could use three land types (three treatments): 1) florally diverse natural land, 2) heterogeneous cropland containing a variety of flowering crop types, including crops depending on bee pollination, as well as non-crop vegetation, and 3) monoculture cropland. We will use a monoculture flowering crop system that relies on honey bee pollination, such as blueberry or cranberry, and will select only fields that are managed without the use of pesticides. We will use ArcGIS to find 5 replicate sites within each of the 3 landcover types for a total of 15 sites in all. Nested within each site we will place 3 honey bee hives in different places. Due to sharing the same landscape, these 3 will not be considered independent replicates, but they will serve to reduce error and produce a better measure of the mean colony health for each landscape. Experimental hives will be developed using 45 identical packages of bees (Italian variety) established into single deep 10 frame hive bodies with drawn comb. After placing hives in early May, we will inspect hives every two weeks until late September. Pollen traps, which collect pollen from honey bee pollen foragers when they enter hives, will be placed on all hives three days prior to inspection and removed on the day of inspection, in order to determine the dominant pollen type brought into each hive. We will assess colony growth and development by weighing hives and measuring proportional brood cover on frames. From within each hive, we will select 20 workers at random to measure and weigh to determine differences in worker size and mass between treatments. We will assess colony health by measuring colony load of two common honey bee pathogens: Nosema ceranae and Varroa destructor mites. Nosema levels will be determined by spore counts of groups of 20 bees (Cantwell 1970) and mite loads will be determined through the use of the powdered sugar method (Macedo et al. 2002). Hypopharyngeal gland protein content of nurse bees will be estimated using Bradford protein assay and colony growth will be measured each week. Phenoloxidase, prophenoloxidase and glucose oxidase enzyme activities will also be analyzed that are indicators of honey bee immune system function (Di Pasquale et al., 2013). The effect of the landscape and thus floral variety treatments on the various outcome measures related to honey bee health will be analyzed with Generalized Linear Mixed Models (for repeated measures) with landscape treatment as a fixed effect and site as a random effect. Pollen collected in pollen traps will be analyzed as both outcome of landscape treatment, and predictor for colony growth and health.Objective 4 -- To assess the effects of exposure to pesticides and other xenobiotics on the survival, health and productivity of honey bee colonies and pollinator abundance and diversitySeed treatment insecticides in agronomic crops: Most of the corn and soybean seed planted in this country has been coated with systemic pesticides. During planting of that seed a dust cloud can be created with very high levels of pesticide which can transport off site and potentially expose bees either directly to the cloud, or, indirectly by landing on bee forage (Krupke et al., 2012).A survey of apiaries will be done that will represent areas of intensive row-crop production, areas of commercial cucurbit vegetable production where honey bees are used for pollination purposes, urban areas, and areas where honey bee exposure to neonicotinoid insecticides is unlikely. Satellite imagery and other land-use databases will be used to assess potential food sources around each apiary.Transport processes affecting seed treatment insecticides will be examined by placing dosimeters at various distances around fields during planting and then analyzing those dosimeters for pesticides by liquid chromatography/mass spectrometry. Different fluency agents added to the seed will be compared to determine practices that will minimize this potential exposure route.Field monitoring for pesticide exposure: An important aspect of assessing the effects of pesticides on bees is to identify routes of exposure and measure concentrations of different pesticides to which the bees would be exposed over time. By trapping pollen as it is brought into the hive by honey bees, collecting it on a regular basis and analyzing it for a range of pesticides using liquid chromatography/mass spectrometry, we can monitor exposure by this route over the long term and quantify "field realistic" levels. We plan to monitor hives in both urban and agricultural environments, and to evaluate toxicity using a Pollen Hazard Quotient (concentration in ppb/LD50 in ug/bee) (Stoner and Eitzer, 2013).Field effects of pesticide exposure: Hives will either receive xenobiotic exposure either through direct application or through direct field exposure when pesticides are used on nearby crops. For direct application exposure the xenobiotics will be applied through contaminated wax sprayed on foundation or contaminated pollen patties or sugar syrup fed to the colonies. Uncontaminated wax, pollen patties or syrup will serve as the control for direct application experiments. For field exposure hives half of the experimental hives will be relocated to the agricultural site(s) at the start of the growing season while half are maintained in areas with less pesticide use.Hives will be given sucrose syrup and protein patties as needed for establishment. All the hive components (box, bottom board, cover and frames with foundation comb), hereafter referred to as the "non-colony" component, will be weighed individually. Bee colonies will be monitored for at least two months prior to deployment in field studies. Research sites in agricultural fields planted with crops that are exposed to at least some bee activity and with moderate to heavy pesticide application will be identified through consultation with growers and beekeepers. During the initial monitoring period, all hives will be kept on stainless steel electronic balances (TEKFA® model B-2418, Galten, Denmark) with an overall precision of ±20 g. Two temperature probes will be placed between the center frames of the brood box. The balance and probes will be linked to 12-bit dataloggers (Hobo® U-12 External Channel, Onset Computer Corp.). The balance and datalogger systems for hives at the agricultural site will be solar- and battery-powered. The dates and locations of pesticide applications and other row crop management practices will be recorded and correlated with changes observed in the continuous datasets. Ambient temperature and rainfall will be recorded throughout the experiments.

Progress 11/28/14 to 09/30/19

Outputs
Target Audience:Presentations and written reports have been made to beekeepers, pesticide applicators and commodity groups representing insect-pollinated crops, specifically almond growers in California. Presentations on Objective 2, exploring the interactive effects of co-applied almond pesticides on bees, was made at the American Beekeeping Federation and the California State Beekeepers Association annual meetings. Similar oral and poster presentations were made at the Almond Board of California meeting where many "Certified Crop Advisors" and others with decision-making authority for pesticide applications were present. Additionally, a research summary was published in the Almond Board's compendium of research updates and findings and a peer-reviewed paper was published on this topic. All stakeholders have been made aware that particular combinations of fungicides and insecticides carry the potential to kill bees even though their constituent pesticides demonstrate low bee toxicity. Objective 4 was accomplished through recommendations based on this research that have been critical to the formulation and recent updates of the Almond Board of California's recommendation in its "Best Management Practices" to avoid insecticide application during almond bloom, to avoid the use of adjuvants and to minimize bee exposure to fungicides through evening application. Changes/Problems: Nothing Reported What opportunities for training and professional development has the project provided?Thirteen undergraduate students from Ohio State, the College of Wooster, and Kent State University were hired and trained to perform bioassay experiments on Varroa mites, larval and adult worker honey bees and honey bee queens over summer 2019. Four of these students continued work on these projects by enrolling for independent research credit and have presented results of their findings at University research conferences as well as national meetings (American Honey Producers Association Annual Conference an American Bee Research Conference). One undergraduate working in the lab has enrolled as a graduate student at OSU and is continuing work to understand the molecular mechanisms underlying synergistic pesticide interactions in bees. How have the results been disseminated to communities of interest?A peer-reviewed paper on synergistic effects in honey bee larvae was published and received wide media attention in California. Additionally, results were presented at the Almond Board of California's annual meeting and a summary was published in this organization's "Research Updates". On the beekeeper side, results were communicated through a presentation at the American Beekeeping Federation meeting, the California State Beekeepers Association meeting and through beekeeper-focused podcasts. What do you plan to do during the next reporting period to accomplish the goals? Nothing Reported

Impacts
What was accomplished under these goals? Ohio State University was involved in Objectives 2-4.Objective 2:Interactive effects between insecticides, fungicides and a spray adjuvant applied to almonds during bloom were explored using a Potter Spray Tower to simulate field spray exposure to adult worker bees. Similar combinations of almond pesticides and an adjuvant were applied to larval worker honey bees in semi-synthetic diet using an in vitro larval rearing protocol. Effects of a subset of almond pesticides on queen development were also tested using standard queen rearing protocols. Both simulated sprays and larval feeding indicate that the combination of the insecticide chlorantraniliprole and the fungicide propiconazole has the potential to kill bees in the field despite the relative safety of these products when applied alone. This synergistic effect between this insecticide and fungicide is increased when an organosilicone-based spray adjuvant is added. Other combinations of insecticides (chlorantraniliprole, methoxyfenozide and diflubenzuron) and fungicides (propiconazole, iprodione, and formulated combinations of boscalid and pyraclostrobin, and fluopyram and trifloxystrobin) did not cause an increase in mortality as substantial as the chlorantraniliprole-propiconazole combination. However, the insecticide diflubenzuron by itself, has demonstrated the potential to kill both larval worker and queen honey bees. Objective 3:A laboratory screening assay was performed testing 12 test compounds for potential use as acaricides to control Varroa destructor in honey bee hives. Assays consisted of applying a coating of test substance on the interior of 20 ml (for mites) or 60 ml (for bees) vials at varying concentration and constructing a dose-response curve for each species. Dose-response curves were then compared to identify the compounds with the greatest "margin of safety" for further development and eventual application in test hives. Objective 4: The potential for harm from insecticides applied to almonds during bloom has been communicated to applicators and almond growers in California and has resulted in a "Best Management Practices" recommendation by the Almond Board of California to avoid insecticide and spray adjuvant application during bloom.

Publications

  • Type: Journal Articles Status: Published Year Published: 2019 Citation: Richardson, R. T., H. R. Curtis, E. G. Matcham, C.-H. Lin, S. Suresh, D. B. Sponsler, L. E. Hearon, and R. M. Johnson. 2019. Quantitative multi-locus metabarcoding and waggle dance interpretation reveal honey bee spring foraging patterns in Midwest agroecosystems. Mol. Ecol. 28: 686697.
  • Type: Journal Articles Status: Published Year Published: 2019 Citation: Wade, A., C.-H. Lin, C. Kurkul, E. R. Regan, and R. M. Johnson. 2019. Combined Toxicity of Insecticides and Fungicides Applied to California Almond Orchards to Honey Bee Larvae and Adults. Insects. 10.
  • Type: Journal Articles Status: Under Review Year Published: 2019 Citation: Richardson, R. T., R. M. Johnson, and D. B. Sponsler. Application of plant metabarcoding to identify diverse honey bee pollen forage in urban ecosystems. Molecular Ecology.
  • Type: Theses/Dissertations Status: Published Year Published: 2018 Citation: Richardson, R. T. 2018. Molecular analysis of honey bee foraging ecology.
  • Type: Journal Articles Status: Submitted Year Published: 2019 Citation: Lin, C.-H., D. B. Sponsler, R. T. Richardson, H. Watters, D. A. Glinski, M. W. Henderson, J. M. Minucci, E. H. Lee, S. T. Purucker, R. M. Johnson , Honey bees and neonicotinoid-treated corn seed: contamination, exposure, and effects. PLoS ONE
  • Type: Journal Articles Status: Published Year Published: 2018 Citation: Johnson, R. M., B. A. Harpur, K. A. Dogantzis, A. Zayed, and M. R. Berenbaum. 2018. Genomic footprint of evolution of eusociality in bees: floral food use and CYPome blooms. Insectes Soc. 65: 445454.


Progress 10/01/17 to 09/30/18

Outputs
Target Audience:Reportshave been made to potentiallyaffected beekeeper groups as well as groups of pesticide applicators and commodity groups for insect-pollinated crops, specifically almond growers in California. A presentation on Objective 1, exploring the interactive effects of co-applied almondpesticides on bees, was made at both theAmerican Honey Producers Association and the American Beekeeping Federation meetings, the two big national beekeeping organizations. A similar presentation was made at the Almond Board of California meeting where many "Certified Crop Advisors" and others with decision-making authority for pesticide applications were present. Additionally, a research summary was published in the Almond Board's compendium of research updates. All stakeholders have been made aware that particular combinations of fungicides and insecticides carry the potential to kill bees even though their constituent pesticides demonstrate low bee toxicity, which has directly addressed Objective 2, which is to reduce the harm to pollinators caused by pesticide applications. Recommendations based on this research have been critical to the formulation of the Almond Board of California's recommendation in its "Best Management Practices" to avoid insecticide application during almond bloom and to minimize bee exposure to fungicides through evening application. Changes/Problems: Nothing Reported What opportunities for training and professional development has the project provided?Eight undergraduate students from Ohio State, the College of Wooster, Unity College and Morehead State University were hired and trained to perform bioassays experiments onVarroamites, and larval and adult worker honey bees over summer 2018. Three of these students continued work on these projects by enrolling for independent research credit and have presented results of their findings at University research conferences as well as regional forums (Ohio Valley Meeting of the Society of Environmental Toxicology and Chemistry) and national meetings (American Bee Research Conference). How have the results been disseminated to communities of interest?Results on pesticides applied to almonds during bloom have been presented at the Almond Board of California annual meetings and have been published in this groups "Research Updates" and a research article is under review summarizing findings for the scientific community. Additionally, findings on almond pesticides have been communicated to national beekeeping groups through invited presentations. General progress onVarroamite work has been communicated to state and regional beekeeping groups through oral presentations. What do you plan to do during the next reporting period to accomplish the goals?In consultation with the Almond Board of California we will change pesticides used in combination testing to better reflect the use of products currently growing in popularity. We are also working to incorporate spray adjuvants, products that alter the rainfastness, longevity or penetration of pesticides, into our laboratory testing. Over the coming year we will perform a simulated spray application of the updated list of pesticides, in field relevant combinations, with and without common spray adjuvants, to determine if these combinations have the potential to kill bees when applied in the field. We will also continue performing laboratory testing on additional novel acaricides onVarroaand adult honey bees in vial bioassays and will determine the products demonstrating the most potential for promotion to field testing. ?

Impacts
What was accomplished under these goals? Objective 2: Interactive effects between insecticides and fungicides applied to almonds during bloom were explored using laboratory bioassays on worker adults and larvae and semi-field assays on honey bee queen development. Adult feeding bioassays, larval feeding bioassays and simulated spray applications to adult bees all indicate that the combination of the insecticide chlorantraniliprole and the fungicide propiconazole has the potential to kill bees in the field despite the relative safety of these products when applied alone. Testing has also looked at other combinations between the insecticides chlorantraniliprole, methoxyfenozide and diflubenzuron with the fungicides propiconazole, iprodione and a formulated combination of boscalid and pyraclostrobin. None of the other combinations has caused increased mortality as substantial as the chlorantraniliprole-propiconazole combination, though the insecticide diflubenzuron, by itself, has demonstrated a potential to kill larval bees. Objective 3: A laboratory screening assay was performed testing 13 different test compounds for their potential utility as acaricides to controlVarroa destructorin honey bee hives. Assays consisted of applying a coating of test substance on the interior of 20 ml (for mites) or 60 ml (for bees) vials at varying concentration and constructing a dose-response curve for each species. Dose-response curves were then compared to identify the compounds with the greatest "margin of safety" for further development and eventual application in test hives. Objective 4: The potential for harm from insecticides applied to almonds during bloom has been communicated to applicators and almond growers in California and has resulted in a recommendation by the Almond Board of California to avoid insecticide applications while almonds are in bloom. Identification of this synergistic combination will also help protect pollinator health when visiting other crops where these very common pesticides may also be co-applied

Publications

  • Type: Journal Articles Status: Awaiting Publication Year Published: 2018 Citation: Richardson, R., Curtis, H., Matcham, E. G., Hua-Lin, C., Suresh, S., Sponsler, D., Hearon, L. Johnson, R. (2018). Quantitative multi-locus metabarcoding and waggle dance interpretation reveal honey bee spring foraging patterns in Midwest agroecosystems. bioRxiv. doi:10.1101/418590
  • Type: Journal Articles Status: Under Review Year Published: 2018 Citation: Wade, A., Lin, C.-H., Kurkul, C.A., Regan, E., Johnson, R.M. (2018) Combined toxicity of insecticides and fungicides applied to California almond orchards to honey bee larvae and adults. Insects.
  • Type: Journal Articles Status: Published Year Published: 2018 Citation: Bengtsson-Palme, J., Richardson, R. T., Meola, M., Wurzbacher, C., Tremblay, �. D., Thorell, K., & Henrik Nilsson, R. (2018). Metaxa2 Database Builder: Enabling taxonomic identification from metagenomic or metabarcoding data using any genetic marker. Bioinformatics . doi:10.1093/bioinformatics/bty482
  • Type: Journal Articles Status: Published Year Published: 2017 Citation: Richardson, R. T., Bengtsson-Palme, J., & Johnson, R. M. (2017). Evaluating and optimizing the performance of software commonly used for the taxonomic classification of DNA metabarcoding sequence data. Molecular Ecology Resources, 17(4), 760769.
  • Type: Conference Papers and Presentations Status: Published Year Published: 2018 Citation: Lin, C. H., Wade, A., Kurkul, C., & Johnson, R. (2018). Effects of insecticide-fungicide combinations commonly applied to almonds during bloom on honey bee larval development. Bee World, 95: 52
  • Type: Conference Papers and Presentations Status: Published Year Published: 2018 Citation: Lin, C. H., Sponsler, D., Richardson, R., Glinski, D., Henderson, M., Minucci, J., Purucker, T., Johnson, R. (2018). Honey bees and neonicotinoid-treated corn seed: Pollen contamination, exposure, and effects. Bee World, 95: 54-55
  • Type: Conference Papers and Presentations Status: Published Year Published: 2018 Citation: Richardson, R., Curtis, H., Lin, C. H., Johnson, R. M. (2018). Investigating honey bee pollen foraging patterns using multi-locus metabarcoding. Bee World, 95: 61
  • Type: Journal Articles Status: Published Year Published: 2018 Citation: Richardson, R. T., Bengtsson-Palme, J., Gardiner, M. M., & Johnson, R. M. (2018). A reference cytochrome c oxidase subunit I database curated for hierarchical classification of arthropod metabarcoding data. PeerJ, 6, e5126.
  • Type: Journal Articles Status: Published Year Published: 2018 Citation: Richardson, R. T., Ballinger, M. N., Qian, F., Christman, J. W., & Johnson, R. M. (2018). Morphological and functional characterization of honey bee, Apis mellifera, hemocyte cell communities. Apidologie, 49(3), 397410.
  • Type: Journal Articles Status: Published Year Published: 2018 Citation: Calla, B., Noble, K., Johnson, R. M., Walden, K. K. O., Schuler, M. A., Robertson, H. M., & Berenbaum, M. R. (2017). Cytochrome P450 diversification and hostplant utilization patterns in specialist and generalist moths: Birth, death and adaptation. Molecular Ecology, 26(21), 60216035.
  • Type: Conference Papers and Presentations Status: Published Year Published: 2018 Citation: Gross, B., & Johnson, R. (2018). Drone brood removal: A bee-utiful form of Varroa control and source of edible insect protein. Bee World, 95: 51


Progress 10/01/16 to 09/30/17

Outputs
Target Audience:The target audiences reached this year have included beekeepers, farmers, and those involved in the registration and testing of pesticides relating to pollinator safety. Almond growers and beekeepers supplying honey bees for almond pollination were a particularly important audience as research on the effect of pesticides applied to almonds during bloom was communicated to these groups. Identification of the effect of insecticides applied to almonds on honey bees has lead to change in the "Best Management Practices" promulgated by the Almond Board of California to include a recommendation that insecticides never be applied during almond bloom. Exposure of honey bees to neonicotinoid insecticide-laden dusts during corn planting continues to be a topic of much concern and presentations on this topic were presented to Canadian and US beekeeping organizations as well as the "Pollinator Community of Practice" at the USEPA. Through interacting with the whole spectrum of stakeholders it is possible to maximize benefit to pollinators by reducing the harm posed by pesticide applications while also addressing the concerns of those potentially harmed by pesticide use. Changes/Problems: Nothing Reported What opportunities for training and professional development has the project provided?During this period eleven undegraduate researchers, four graduate students and a postdoctoral research associate have worked together to design, execute, analyze and publish pollinator toxicology and related research. Methods developed for research using pollen analysis, honey bee colony modeling and honey bee waggle dance analysis have been modified to serve as laboratory exercises in OSU's course in Beekeeping. Lab members and affiliates participated in a free screening for Varroa mites a at the Fall meeting of the Ohio State Beekeepers Association where beekeepers both received health results on-site and had the opportunity to participate in this analysis. How have the results been disseminated to communities of interest?Research findings have been reported in peer-reviewed journals and have been published in a thesis and dissertation. A number of research talks have been presented at meetings of scientific societies. Additionally, several lab members have communicated findings directly to beekeeper groups through invited talks at local, state, regional and international beekeeper meetings as well as through the monthly OSU Bee Lab webinar series.Findings related to regulatory toxicology have been delivered to US EPA staff through a series of special webinars to the "Pollinator Community of Practice" and through an in-person at both a short course at the Society for Environmental Toxicology and the Chemistry annual meeting. Finally, the effects of insecticide-fungicide combinations have been communicated to almond growers through a poster presentation and talk presented at the Almond Board of California meeting and through communication with the Almond Board. What do you plan to do during the next reporting period to accomplish the goals?Insecticide-fungicide combinations used on almonds during bloom will be tested with the addition of spray adjuvants to determine the combinatorial effect on adult bee survival and larval bee development (Objective 2). A multi-state collaboration to identify new chemistries with promise for controlling Varroa mites in honey bee hives has been assembled and will begin testing a variety of compounds in 2018 (Objective 2 and 4). Finally, work has begun looking at the potentially beneficial effects of soybean production on honey bee honey production through a collaboration with Ohio beekeepers (Objectives 1 and 4).

Impacts
What was accomplished under these goals? Objective 1 and 2: Based on an analysis of insecticides and fungicides, and their combinations, applied to almond orchards during bloom we continued testing the effects of common pesticides and combinations to different castes and life stages of honey bees. This involved chronic feeding of adult honey bees with pollen containing almond pesticide mixtures as well as feeding worker and queen larvae these same compounds. Results to date suggest that in 2015 approximately 50,000 honey bee colonies used for almond pollination may have been exposed to insecticides or insecticide-fungicide combinations with the potential to negatively affect both larval development and adult survival. Objective 3 and 4: Varroa mite infestations, combined with the viruses vectored by Varroa mites, continue to be a very serious problem facing beekeepers in the U.S. in the previous year a multi-state meeting of researchers, beekeepers and other stakeholders was convened with the goal of submitting a multi-institutional USDA-SCRI Grant to fund work to solve the Varroa problem. Additionally, new products with potential for Varroa control have been screened in the laboratory for mite toxicity and bee safety. One of these compounds was tested in a limited field trial. Objective 2: The release of insecticide-containing dust derived from neonicotinoid-treated corn seeds during planting continues to cause sporadic bee kills in Ohio and other corn-belt states. We have completed pesticide residue analysis on bees and hive matrices collected as part of a three-year study and incorporated them into a model of insecticide dust drift and bee exposure potential. Analysis of these data indicate that there are no easy routes to landscape-level or beekeeping-related mitigation of insecticide emission and that mitigation effort should be focused on increasing the adherence of insecticide to the seed and capturing or directing the movement of any dust that is produced.

Publications

  • Type: Theses/Dissertations Status: Published Year Published: 2017 Citation: Riusech, N. S., R. M. Johnson, Environmental Science Graduate Program., and Ohio State University. 2017. Varroa mite control in honey bee colonies : he use of a fatty acid blend (C8910) for Varroa mite control and exploring management practices used by beekeepers in full-sized colonies.
  • Type: Journal Articles Status: Published Year Published: 2017 Citation: Sponsler, D. B., and R. M. Johnson. 2017. Poisoning a Society: A Superorganism Perspective on Honey Bee Toxicology. Bee World. 94: 3032.
  • Type: Journal Articles Status: Published Year Published: 2017 Citation: Sponsler, D. B., E. G. Matcham, C.-H. Lin, J. L. Lanterman, and R. M. Johnson. 2017. Spatial and taxonomic patterns of honey bee foraging: A choice test between urban and agricultural landscapes. J Urban Ecol. 3.
  • Type: Journal Articles Status: Published Year Published: 2017 Citation: Sponsler, D. B., and R. M. Johnson. 2017. Mechanistic modeling of pesticide exposure: The missing keystone of honey bee toxicology. Environ. Toxicol. Chem. 36: 871881.
  • Type: Theses/Dissertations Status: Published Year Published: 2016 Citation: Sponsler, D. B., R. M. Johnson, Department of Entomology., and Ohio State University. 2016. Honey bee landscape ecology : foraging, toxic exposure, and apicultural outcomes.
  • Type: Journal Articles Status: Published Year Published: 2017 Citation: Richardson, R. T., J. Bengtsson-Palme, and R. M. Johnson. 2016. Evaluating and optimizing the performance of software commonly used for the taxonomic classification of DNA metabarcoding sequence data. Mol. Ecol. Resour.
  • Type: Journal Articles Status: Published Year Published: 2017 Citation: Calla, B., K. Noble, R. M. Johnson, K. K. O. Walden, M. A. Schuler, H. M. Robertson, and M. R. Berenbaum. 2017. Cytochrome P450 diversification and hostplant utilization patterns in specialist and generalist moths: Birth, death and adaptation. Mol. Ecol.
  • Type: Journal Articles Status: Published Year Published: 2017 Citation: Swale, D. R., D. W. Engers, S. R. Bollinger, A. Gross, E. A. Inocente, E. Days, F. Kanga, R. M. Johnson, L. Yang, J. R. Bloomquist, C. R. Hopkins, P. M. Piermarini, and J. S. Denton. 2016. An insecticide resistance-breaking mosquitocide targeting inward rectifier potassium channels in vectors of Zika virus and malaria. Sci. Rep. 6: 36954.


Progress 10/01/15 to 09/30/16

Outputs
Target Audience:The target audiences this year have included beekeepers, gardeners, and farmers, as well as those involved in the registration and testing of pesticides relating to pollinator safety. Exposure of honey bees to neonicotinoid insecticide-laded dusts during corn planting continues to be a topic of much concern and presentations on this topic were presented to state and provincial beekeeping organizations in the U.S. and Canada as well as CropLife America and the USEPA. Research on the effects of pesticide combinations is another aspect of our research and these results have been presented to growers (The Almond Board of California) and to beekeepers and regulators. Through interactions with the whole spectrum of stakeholders it is possible to maximize benefit to pollinators by reducing the harm posed by pesticide applications while also addressing the concerns of those potentially harmed by pesticide use. Changes/Problems: Nothing Reported What opportunities for training and professional development has the project provided?During this period six undegraduate researchers, three graduate students and a postdoctoral research associate have worked together to design, execute, analyze and publish pollinator toxicology and related research. Methods developed for honey bee dance analysis and pollen identification though the course of research have formed the basis for undergraduate laboratory exercises in OSU's course in Beekeeping. Additionally, all lab members provided a live free screening session for Nosema, a gut parasite of bees, at the Fall meeting of the Ohio State Beekeepers Association where beekeepers both received health results on-site and had the opportunity to participate in this analysis. How have the results been disseminated to communities of interest?Research findings have been reported in peer-reviewed journals. Additionally, most lab members, including a postdoctoroal researcher and graduate students, have communicated findings to beekeeper groups through a number of invited talks delivered at local, state, and national beekeeper meetings as well as the OSU Bee Lab webinar series. Findings and processes related to regulatory toxicology have been delivered to US EPA staff through a series of special webinars and presented in-person at both a short course at the Society for Environmental Toxicology and Chemistry annual meeting.Finally, the effects of insecticide-fungicide combinations have been communicated to almond growers through a poster presentation at the Almond Board of California meeting and through communication with the Almond Board. What do you plan to do during the next reporting period to accomplish the goals?Additional insecticide-fungicide combinations used on almonds during bloom will be explored and effects on adults and developing honey bees will be documented and shared (Objective 2). We anticipate identifying the most desired arenas of Varroa mite and virus research at the upcoming USDA-SCRI Planning meeting and will work to both generate preliminary data and seek funding for this research priority (Objective 4).

Impacts
What was accomplished under these goals? Objective 2. An analysis of insecticide-fungicide combinations applied to almond orchards during bloom was conducted using the California Pesticide Use database. The most widely used combinations were applied to both adults and queens to assess the effects that bloom-time tank-mix sprays may have on the approximately 1.8 million honey bee colonies rented for almond pollination. Objective 2: Release of neonicotinoid dust during the process of planting insecticide-coated corn seed continues to cause sporadic bee kills in Ohio and other corn-belt states. Three years of data have been collected from Ohio apiaries during corn planting as well as dust drift data. Analysis of these data has documented the scale of the problem posed by insecticidal corn seed treatment dust and, using a variety of modeling approaches, are beginning to point toward control of flowering weeds in corn fields as potentially the most feasible mitigation option. Objective 4. The Varroa mite, and the viruses it vectors, continues to be one of the most serious problems facing beekeepers in the US. Recognizing that new approaches to this problem are needed a multi-state meeting of researchers (including other participants in NC1173), beekeepers and other stakeholders was organized and funded with a USDA-SCRI Planning Grant. The meeting will examine the efficacy, efficiency and other factors regarding current Varroa mite approaches and identify areas of mite control and monitoring that would be especially fruitful areas of research in the future.

Publications

  • Type: Journal Articles Status: Published Year Published: 2015 Citation: Richardson, R. T., C.-H. Lin, J. O. Quijia, N. S. Riusech, K. Goodell, and R. M. Johnson. 2015. Rank-Based Characterization of Pollen Assemblages Collected by Honey Bees Using a Multi-Locus Metabarcoding Approach. Appl. Plant Sci. 3: 1500043.
  • Type: Journal Articles Status: Awaiting Publication Year Published: 2016 Citation: Sponsler, D. B., and R. M. Johnson. 2016. Mechanistic modeling of pesticide exposure: the missing keystone of honey bee toxicology. Env. Tox. Chem. DOI: 10.1002/etc.3661
  • Type: Journal Articles Status: Awaiting Publication Year Published: 2016 Citation: Richardson, R. T., Bengtsson-Palme, J., and R. M. Johnson. 2016. Evaluating and optimizing the performance of software commonly used for the taxonomic classification of DNA metabarcoding sequence data. Mol. Ecol. Res. DOI: 10.1111/1755-0998.12628


Progress 11/28/14 to 09/30/15

Outputs
Target Audience:In the past year the target audiences for efforts have included beekeepers; corn, vegetableand almond farmers; agricultural equipment manufacturerers; seed dealers; agrochemical companies; regulators and legislators. The interface between pesticides and honey bees continues to be a topic of intense interest to many diverse stakeholder groups and we have worked to communicate findings to these audiences through formal talks and written publications. Exposure of honey bees to neonicotinoid insecticides during the process of corn planting is a topic of much interest in Ohio and other corn-belt states. Presentations have been made to beekeeping groups, agribusiness organizations and a national meeting of state legislators on this topic. Short articles have also been written on this topic for publication in the Ohio State Beekeepers Association publications as well as university-produced articles aimed at corn farmers. Awareness of the problem has been raised among these groups and solutions t the corn seed treatment dust problem are under consideration. Pesticide application to almonds, the largest and most valuable crop pollinated by honey bees, isof great concern to beekeepers and presentations have been made on this topic to both beekeeping groups and almond growers in California. These efforts have raised awareness among these groups of the prevalence of insecticide application to these crops during almond bloom. Changes/Problems:The regulatory landscape relating to pesticides and bees has undergone major changes since the project was initiated. Regulators are no longer relying on adult acute toxicity measurements to assess the risk of pesticides -- testing on larval and chronic toxicity are now being required as well. Over the course of the coming year I will work to incorporate these additional tests into my research as well as continue to develop new methods and new endpoints further upon the "Adverse Outcome Pathway".? What opportunities for training and professional development has the project provided?Three graduate students have been involved in assessing the exposure and effects of seed treatment insecticides on honey bee colonies as has one postdoctoral research associate. These students and postdoc have presented their findings through 3 written publications and oral and poster presentations at national meetings. Two of these students have transitioned from the Master's program to the Ph.D. program in entomology. Additionally, 5 undergraduates at Ohio State University and Cedarville University have contributed to the research and twohave taken the opportunity to present their findings at a University research symposium. Two high-school students have also been involved in lab work identifying pollen collected by bees in Ohio. How have the results been disseminated to communities of interest?Results have been presented at national, state and local beekeeper meetings in Ohio and California to beekeepers that are potentially affected by the pesticide exposure in corn, cucurbits and almonds. Presentations have also been made to farmer and agribusiness groups for corn, cucurbitsand almonds. A series of articles for beekeepers have also been published in the Ohio State Beekeepers' Association newsletter. Pesticide issues are also of interest to regulators in government and legislators, and presentations have been made to government workers and state-level legislators.? What do you plan to do during the next reporting period to accomplish the goals?SpecificObjective 2: We plan to deterimine the colony-level effect of bee exposure to corn seed treatment dust by following and assessing colonies through the year following exposure during corn planting. Experiments comparing honey bees and squash bee susceptibility to insecticides and insecticide-fungicide combinations are also ongoing. Specific Objective 4: Recommendations will be made to almond, cucurbit and corn grower groups on insecticide and fungicide applications that may harm bees and ways that these effects could be mitigated.

Impacts
What was accomplished under these goals? Specific Objective 2: The effects of pesticides applied to cucurbit crops, including common combinations of insecticides and fungicides, were tested on two species of bees that are important for pollination of these cucurbit crops -- the managed honey bee and the wild squash bee. Preliminary results indicate that there are significant differences in the toxicity of insecticides and combinations to these two vital pollinators of squash and melon crops. Specific Objective 2: Bee exposure and mortality associated with neonicotinoid insecticides applied to corn as a seed treatment, through dust-off, was measured at apiaries around central Ohio. Bees reliably returned to the hive with seed treatment insecticides in pollen during the corn planting period and the appearance of these insecticides was associated with an increase in the number of dead bees ejected by colonies. Specific Objective 4:Changes in management of in-field blooming weeds could be an easy way to reduce bee exposure to insecticides in corn seed treatment dust.

Publications

  • Type: Journal Articles Status: Published Year Published: 2015 Citation: Sadd, B. M., S. M. Barribeau, G. Bloch, D. C. de Graaf, P. Dearden, C. G. Elsik, J. Gadau, C. J. Grimmelikhuijzen, M. Hasselmann, et al. 2015. The genomes of two key bumblebee species with primitive eusocial organization. Genome Biol. 16: 76.
  • Type: Journal Articles Status: Published Year Published: 2015 Citation: Sponsler D.B. & Johnson R.M. 2015. Honey bee success predicted by landscape composition in Ohio, USA. PeerJ. 3:e838. doi: 10.7717/peerj.838
  • Type: Journal Articles Status: Published Year Published: 2015 Citation: Richardson, R.T., Lin, C.-H., Quijia Pillajo, J.O., Sponsler, D.B., Goodell, K., Johnson, R.M. 2015. Application of ITS2 metabarcoding to determine the provenance of pollen collected by honey bees in a field-crop dominated agroecosystem. Applications in Plant Sciences. 3 (1): 1400066 doi: 10.3732/apps.1400066
  • Type: Journal Articles Status: Published Year Published: 2015 Citation: Johnson, R.M. 2015. Honey bee toxicology. Annual Review of Entomology. 60:415-34. doi: 10.1146/annurev-ento-011613-162005
  • Type: Journal Articles Status: Published Year Published: 2015 Citation: Kapheim KM, Pan H, Li C, Salzberg SL, Puiu D, Magoc T, Robertson HM, Hudson ME, Venkat A, Fischman BJ, et al. 2015. Social evolution. Genomic signatures of evolutionary transitions from solitary to group living. Science. 348:11391143.
  • Type: Journal Articles Status: Published Year Published: 2015 Citation: Berenbaum MR, Johnson RM. 2015. Xenobiotic detoxification pathways in honey bees. Current Opinion in Insect Science. 10:5158.
  • Type: Journal Articles Status: Published Year Published: 2014 Citation: Akinwande, K.L , Dahlgren, L.P., Johnson, R.M., Siegfried, B.D. and Ellis, M.D. 2014. Effect of amitraz on queen honey bee egg and brood development. Mellifera. 14(26-27):33-40