Progress 04/01/18 to 03/31/22
Outputs
Target Audience:This work is focussed on control strategies for a major honey bee pathogen and the targets are federal and university scientists, industry members, and ultimately the beekeeping stakeholders Changes/Problems:We were strongly affected by COVI-19 restrictions in the Lab in 2020, but the PDs were able to come in and Dr. Ryabov was able to enact the viral marker experiments and RNAi as planned. Others were funded to help develop viral genetic resources and were prepared as restrictions eased to finish experiments during the one-year extension. What opportunities for training and professional development has the project provided?The project trained and employed researchers at all levels from high school interns (5 total in four years) to undergraduate and graduate students, and four visiting postdoctoral fellows (two from the US, one from the UK, and one from Spain). Three of these individuals have moved to additional jobs in science and industry, while one (Evan Palmer Young) continues to do science with USDA, albeit on a different project. We have helped undergraduates continue for advanced degrees. All lab members take required safety and ethics training, and acquire skills in both fieldwork (beekeeping) and lab work from controlled inoculations to genetic screens for viruses. How have the results been disseminated to communities of interest?Yes, the PD and co-PD Chen deliver annual talks to ten beekeeping groups and the national stakeholder meetings (Virtual in 2021 and 2022). We also regularly converse with beekee[ers and other stakeholders to address and mitigate viral stress on honey bees. We have published nine papers (below) under this project and proposed one patent (for detection of honey bee viruses). Evans JD, Banmeke O, Palmer-Young EC, Chen Y & Ryabov EV (2022) Beeporter: Tools for high-throughput analyses of pollinator-virus infections. Molecular Ecology Resources 22: 978-987. doi:10.1111/1755-0998.13526. Huang S, Li J, Zhang Y, Li Z, Evans JD, Rose R, Gilligan TM, LeBrun A, He N, Zheng T, Zhang T, Hamilton M & Chen YP (2021) A novel method for the detection and diagnosis of virus infections in honey bees. Journal of Virological Methods 293. doi:10.1016/j.jviromet.2021.114163. Posada-Florez F, Childers AK, Heerman MC, Egekwu NI, Cook SC, Chen Y, Evans JD & Ryabov EV (2019) Deformed wing virus type A, a major honey bee pathogen, is vectored by the mite Varroa destructor in a non-propagative manner. Scientific Reports 9. doi:10.1038/s41598-019-47447-3. Posada-Florez F, Lamas ZS, Hawthorne DJ, Chen Y, Evans JD & Ryabov EV (2021) Pupal cannibalism by worker honey bees contributes to the spread of deformed wing virus. Scientific reports 11: 8989. doi:10.1038/s41598-021-88649-y. Posada-Florez F, Ryabov EV, Heerman MC, Chen Y, Evans JD, Sonenshine DE & Cook SC (2020) Varroa destructor mites vector and transmit pathogenic honey bee viruses acquired from an artificial diet. PLoS ONE 15. doi:10.1371/journal.pone.0242688. Ryabov EV, Childers AK, Lopez D, Grubbs K, Posada-Florez F, Weaver D, Girten W, vanEngelsdorp D, Chen Y & Evans JD (2019) Dynamic evolution in the key honey bee pathogen deformed wing virus: Novel insights into virulence and competition using reverse genetics. PLoS Biology 17. doi:10.1371/journal.pbio.3000502. Ryabov EV, Christmon K, Heerman MC, Posada-Florez F, Harrison RL, Chen Y & Evans JD (2020) Development of a honey bee RNA virus vector based on the genome of a deformed wing virus. Viruses 12. doi:10.3390/v12040374. Weaver DB, Cantarel BL, Elsik CG, Boncristiani DL & Evans JD (2021) Multi-tiered analyses of honey bees that resist or succumb to parasitic mites and viruses. BMC Genomics 22. doi:10.1186/s12864-021-08032-z. Zhao Y, Heerman M, Peng W, Evans JD, Rose R, Degrandi-Hoffman G, Simone-Finstrom M, Li J, Li Z, Cook SC, Su S, Rodríguez-García C, Banmeke O, Hamilton M & Chen Y (2019) The dynamics of Deformed wing virus concentration and host defensive gene expression after varroa mite parasitism in honey bees, Apis mellifera. Insects 10. doi:10.3390/insects10010016. What do you plan to do during the next reporting period to accomplish the goals?
Nothing Reported
Impacts
What was accomplished under these goals?
1) We greatly improved the understanding of when RNA interference works and when it fails to protect honey bees from their key viruses. This control strategy will be used to develop products for beekeepers and several university and industry groups are building on data from our efforts and other recent efforts. 2) We found a surprising result (Ryabov et al., below) that actually there is no predominance by particular Deformed wing virus strains in coinfections nor was there a 'protective' effect among the current resident strains. INstead there seems to be RNAi-based selection for rare strains and those strains might fare better, keeping viral diversity high Ryabov EV, Childers AK, Lopez D, Grubbs K, Posada-Florez F, Weaver D, Girten W, vanEngelsdorp D, Chen Y & Evans JD (2019) Dynamic evolution in the key honey bee pathogen deformed wing virus: Novel insights into virulence and competition using reverse genetics. PLoS Biology 17. doi:10.1371/journal.pbio.3000502.
Publications
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Progress 04/01/20 to 03/31/21
Outputs
Target Audience:The scientists produced peer-reviewed publications in this rating period as well as delivered Zoom or Teams-based talks to the American Honey Producers Association, theAmerican Beekeeping Federation, and four state beekeeping groups. They also aided the community of science by sharing reagents and know-how for marker-tagged Deformed wing virus. Changes/Problems:We were almost completely locked out of laboratories for the spring of 2020, beginning again in late summer. We worked on analyzing datasets from the previous year and one from several years before related to this virus and bee hosts. These manuscripts have been submitted and are in review. What opportunities for training and professional development has the project provided?Drs, Eugene Ryabov and Yi Zhang were given training that ultimately will lead to independent positions and an improvement of science. How have the results been disseminated to communities of interest?We have given multiple university and society seminars related to this work and have also made the sequences and clones used freely available to science colleagues. We are often asked by beekeepers what is new and in production and keep them informed of progress and setbacks. What do you plan to do during the next reporting period to accomplish the goals?We will imp[rove the use of Deformed wing virus as a toolfor understanding infection and honey bee defenses, and we will assay the effects of immune stimulation on bee responses to this virus.
Impacts
What was accomplished under these goals?
We finalized a system for generating infectious clones of multiple Deformed wing virus strains., speeding the search for new drug treatments and allowing studies to seehow RNA interference impacts virus infection.
Publications
- Type:
Journal Articles
Status:
Published
Year Published:
2021
Citation:
Huang S, Li J, Zhang Y, Li Z, Evans JD, Rose R, Gilligan TM, LeBrun A, He N, Zheng T, Zhang T, Hamilton M & Chen YP (2021) A novel method for the detection and diagnosis of virus infections in honey bees. Journal of Virological Methods 293. doi:10.1016/j.jviromet.2021.114163.
- Type:
Journal Articles
Status:
Published
Year Published:
2021
Citation:
Posada-Florez F, Lamas ZS, Hawthorne DJ, Chen Y, Evans JD & Ryabov EV (2021) Pupal cannibalism by worker honey bees contributes to the spread of deformed wing virus. Scientific reports 11: 8989. doi:10.1038/s41598-021-88649-y.
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Progress 04/01/19 to 03/31/20
Outputs
Target Audience:Progress reports were given in a total of 11 talks to stakeholder (beekeeping industry) groups. Four talks were given at scientific meetings to entomologists and geneticists. Three papers were published on the research topic of this grant in peer-reviewed journals. Changes/Problems:This virus is the most common one in US honey bees and we are challenged to find clean bees for our inoculation experiments. We have found one commercial beekeeper with exceptionally clean bees and hope to work with them to develop better sources for our experiments. Generally, our biggest need has been professional expertise to help analyze RNA sequences and carry out technical virus assays. What opportunities for training and professional development has the project provided?Along with Dr. Ryabov (visiting postdoctoral scientist), the laboratory was able to employ one postgraduate researcher (David Saleh), and two undergraduate interns, along with three volunteer interns. All were able to take part in laboratory and field work with bees and viruses. How have the results been disseminated to communities of interest?We regularly present results form this work and other laboratory projects at County and State beekeeping meetings. We were not able to attend the two national beekeeping meetings in January 2019, due to the government shutdown, but we are presenting at these meetings in January2020. We have also presented at a USDA_NASA STEM event for students under-represented in science, and have host two High School students from under-represented communities. What do you plan to do during the next reporting period to accomplish the goals?Right now we are investigating the abilities of this virus to escape RNA interference in honey bees, by further tests using specific virus clones. Weare also exploring virus and cell dynamics in honey bees and mites. We have recently developed an infectious clone for DWV-B, a relatively recent, and virulent, strain of this virus that is now in the US. We can now conduct side by side tests to see how these viruses compete, their relative impacts on bees, and the risks ofrecombination in helping viruses escape bee immunity.
Impacts
What was accomplished under these goals?
We carried out RNA sequencing directed at identifying honey bee immune responses and virus counter-responses, using strain-specific virus clones. The results from these extensive experiments are being prepared for publication. Also in preparation is a description of flourescent labels (GFP) in this virus in hopes this will give new details into where viruses travel in bees and parasitic Varroa mites (a vector of Deformed wing virus)
Publications
- Type:
Journal Articles
Status:
Published
Year Published:
2019
Citation:
Posada-Florez, F.; Childers, A. K.; Heerman, M. C.; Egekwu, N. I.; Cook, S. C.; Chen, Y.; Evans, J. D.; Ryabov, E. V., Deformed wing virus type A, a major honey bee pathogen, is vectored by the mite Varroa destructor in a non-propagative manner. Scientific Reports 2019, 9 (1).
- Type:
Journal Articles
Status:
Published
Year Published:
2019
Citation:
Huang, Q.; Lopez, D.; Evans, J. D., Shared and unique microbes between Small hive beetles (Aethina tumida) and their honey bee hosts. MicrobiologyOpen 2019, 8 (10).
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Progress 04/01/18 to 03/31/19
Outputs
Target Audience:We are conducting this work in order to develop new control methods for honey bee viruses. We met with stakeholders at six state-level beekeeping meetings and delivered presentation materials for the American Honey Producers Association and American Beekeeping Association meetings (could not attend either due to US government furlough). PD Evans wrote a monthly research review column for Bee Culture magazine and was able to sue this to advertise the impacts of bee viruses. Changes/Problems:So far, we have been able to meet the grant goals and are now focussing on disseminating new results. What opportunities for training and professional development has the project provided?CoPI Ryabov (an International Visitor to the USDA via the US Forest Service received training in bioinformatics and bee immunity. Two high school students and three 180-term employees employed by the laboratory also gained training as part of this project, along with the 180-day employee (Vy Nguyen) employed by the grant funding. How have the results been disseminated to communities of interest?We have given seminars at the University of Florida, the Cold Spring Harbor bLabs, the Entomological Society of America, and numerous beekeeping stakeholder groups regarding this work, and are also seeking industry collaborators in order to develop new bee medicines. What do you plan to do during the next reporting period to accomplish the goals?We are carrying out competition experiments among virus strains and knocking down specific strains using RNA interference. We are also labelling the infectious clones with fluorescent markers to better understand how they impact bees, and are planning to expand our testing and use of novel antiviral drugs.
Impacts
What was accomplished under these goals?
Infectious clones were developed for Deformed wing virus, a first for the US and a tool that will enable direct testing of antiviral strategies as well as a more full understanding of strain virulence in these viruses. These clones were used in an initial test of infectivity of honey bees by pollen. Finished experiments have shown equally competitive abilities of local strains and a suggestion that strains undergo negative frequency-dependent selection, perhaps because of pressures from the bee immune response. The team also carried out novel work determining how varroa mites interact with Deformed wing virus.
Publications
- Type:
Journal Articles
Status:
Awaiting Publication
Year Published:
2019
Citation:
Eugene V. Ryabov, Anna K. Childers, Dawn Lopez, Kyle Grubbs, Francisco Posada-Florez, Daniel Weaver, Dennis vanEngelsdorp, Yan Ping Chen, Jay D. Evans (2019) Dynamic evolution in the key honey bee pathogen Deformed wing virus, PLoS Biology, in review
- Type:
Journal Articles
Status:
Published
Year Published:
2018
Citation:
Stamets, P. E., et al. (2018) Extracts of Polypore Mushroom Mycelia Reduce Viruses in Honey Bees. Scientific Reports. 8,
- Type:
Journal Articles
Status:
Published
Year Published:
2019
Citation:
2. Maori, E., Navarro, I. C., Boncristiani, H., Seilly, D. J., Rudolph, K. L. M., Sapetschnig, A., Lin, C. C., Ladbury, J. E., Evans, J. D., Heeney, J. L., Miska, E. A. (2019) A Secreted RNA Binding Protein Forms RNA-Stabilizing Granules in the Honeybee Royal Jelly. Molecular Cell. 74, 598-608.e6
- Type:
Journal Articles
Status:
Published
Year Published:
2019
Citation:
Zhao, Y., et al. (2019) The dynamics of Deformed wing virus concentration and host defensive gene expression after varroa mite parasitism in honey bees, Apis mellifera. Insects. 10
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