GENETICS, SELECTION AND BREEDING OF NEW WORLD HONEY BEE POPULATIONS AND COLONY HEALTH

• Sponsoring Institution: National Institute of Food and Agriculture
• Project Status: COMPLETE
• Funding Source: HATCH
• Reporting Frequency: Annual
• Accession No.: 1007314
• Project Start Date: Aug 11, 2015
• Project End Date: Mar 31, 2019
• Program Code: [(N/A)]- (N/A)

Recipient Organization
WASHINGTON STATE UNIVERSITY
240 FRENCH ADMINISTRATION BLDG
PULLMAN,WA 99164-0001

Performing Department
Entomology

Non Technical Summary
Honey bees face a number of challenges to maintain "colony health". These include: aspects of modern agricultural production (e.g - monocultures set in chemically mediated "weed-free" areas), land use patterns by humans that reduce bee forage, the historical context and genetic consequences of the introduction of the honey bee as a "new insect" into North America and a variety of recently introduced parasites and diseases. The focus of this project is to address a subset of these challenges through genetics, in part by importing additional honey bee genetic diversity to provide raw material for selective breeding. Selective breeding of honey bees for parasite and disease tolerance in this project provides the potential for a more sustainable future for crop pollination than continued reliance on synthetic pesticides or antibiotics in beehives. Associated with these efforts is development of practical methods of semen cryopreservation in the honey bee, making possible a functional Honey Bee Germplasm Respository (HBGR). Reconsideration of differences among honey bee subspecies, derived from adapation to different ecological and climatic conditions, allows beekeepers the means to select honey bees strains adapted for their specific geographic needs. Evaluation of these differences has only recently been possible through semen importation and cryopreservation.This Project also explores alternative approaches to improve honey bee colony health, including evaluation of procedures to manipulate metabolic gases in hives placed in controlled storage to control mites and reduce food consumption, evaluation of the anti-viral and anti-mite properties of mycological and other biological-sourced extracts in hives and cultural and management approaches to support colony health.

Animal Health Component

50%

Research Effort Categories

Basic

25%

Applied

50%

Developmental

25%

Classification

Knowledge Area (KA)	Subject of Investigation (SOI)	Field of Science (FOS)	Percent
211	3010	1080	35%
216	3010	1081	35%
303	3099	1070	30%

Knowledge Area
303 - Genetic Improvement of Animals; 211 - Insects, Mites, and Other Arthropods Affecting Plants; 216 - Integrated Pest Management Systems;

Subject Of Investigation
3099 - Bees, honey, and other pollinators, general; 3010 - Honey bees;

Field Of Science
1070 - Ecology; 1081 - Breeding; 1080 - Genetics;

Keywords

honey bee

cryopreservation

genetic diversity

breeding

varroa mite

colony health

Goals / Objectives
The objectives of this project are:1) to examine the genetic relationships among Old World subspecies and New World populations of honey bees, import appropriate germplasm to augment US populationsand develop a research program on improved cryopreservation of honey bee germplasm for breeding and conservation purposes;2) to continue an ongoing bee breeding program that incorporates selection criteria compatible with an integrated pest management approach to control parasites and pathogens;3) to conduct a research program on honey bee colony health related to improvements in indoor wintering protocols, including metabolic gas manipulation, and evaluate alternative parasite control strategies using mycological and other bio pesticides;4) conduct research on basic mechanisms of genetic differentiation in honey bees that will have application to the molecular systematics of the genus (Apis) and insect biosystematic research in general.

Project Methods
The evaluation of these efforts will take place in the form of scientific publications.Various findings of the project have high potential to be more immediately "applied" to existing problems and can thus improve processes and management on a short term timescale. These findings will be disseminated through workshops, regional and national beekeeper and grower association meeting and popular press articles. Evaluation of the impacts of these findings will be measureable through:1) adoption of new genetic strains by the queen producer industry and their customers. The reintroduction of one or more novel strains derived from subspecies will be measureable by stakeholder acceptance and willingness to "change" from currently used honey bees. Similarly, the evaluation of our germplasm introduction efforts will be measureable by the degree of utilization of Project-supplied breeding stock by US commercial queen producers.2) the adoption of recommended management processes (metabolic gases - controlled atmosphere storage), Milestones indicating change would include construction of indoor and controlled atmosphere storage facilities for bees or modification of existing units for such purposes.3) utilization of alternative mite control products (bio sources, mycological). Continued movement away from the use of synthetic pesticides by beekeepers and "toward" the use of biopesticides and mycologically based adjuvants to colony health will be an indicator of success.4) continued growth in the number of contributions to and the utilization of honey bee cryopreservation and the Honey Bee Germplasm Repository (HBGR). 5) continued submission of beekeeper-derived samples from the Pacific Northwest to the Honey Bee Diagnostic laboratory.

Progress 08/11/15 to 03/31/19

Outputs
Target Audience:beekeepers, growers, honey bee queen producers, scientists Changes/Problems: Nothing Reported What opportunities for training and professional development has the project provided?Two postdoctoral researchers continued to be supported and involved in fungal /honey bee research - plant pathology/entomology interdisciplinary projects Three graduate students are involved in this research How have the results been disseminated to communities of interest?Research findings have been disseminated at numerous bee research and regional beekeeper meetings, (CA,ID,OR,WA) and at national meetings (American Beekeeping Federation, American Honey Producers Association) What do you plan to do during the next reporting period to accomplish the goals? Nothing Reported

Impacts
What was accomplished under these goals? Honey bee breeder queens derived from imported honey bee semen were produced and supplied to western honey bee queen producers Evaluation of colony longevity and health parameters form the "stationary apiary" experiment continued in 2018 Experimental testing of fungal extracts for viral reduction in honey bees was tested in a large scale field experiment in California Indoor wintering and storage effects on honey bee physiology and colony strength in commercail storage facilities were evaluated

Publications

• Type: Journal Articles Status: Published Year Published: 2018 Citation: Stamets, P., Naeger, N., Evans, J., Han, J., Hopkins, B., Lopez, D, Moershel, H., Nally, R., Sumerlin, D., Taylor, A., Carris, L., Sheppard, W. 2018. Extracts of polypore mushroom mycelia reduce viruses in honey bees. Scientific Reports, 8:13936. https://doi.org/10.1038/s41598-018-32194-8

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

Outputs
Target Audience:Beekeepers, growers, honey bee queen producers, scientists Changes/Problems: Nothing Reported What opportunities for training and professional development has the project provided?Two postdoctoral researchers continue to be involved in the fungal/honey bee research and have been working both with entomologists and plant pathologists in an interdisciplonary fashion. 3 graduate students are involved in research aspects of colony health experiments and reproductive biology of the honey bee. How have the results been disseminated to communities of interest?Research results from the fungal work, the bee breeding and germplasm improvement work and teh indoor wintering metabolic gas work have been disseminated at numerous meetings of local and region beekeepers (OR, WA, ID, CA, NE, MT,AL) and at national venues, including the Entomological Society of America National meeting and the American Beekeeping Federation National meeting. What do you plan to do during the next reporting period to accomplish the goals?Research results from a large scale field study in CA and OR, and from the stationary apiary experiment that ended in 2018 are currently being analyzed and will be reported in peer-reviewed journals during the next reporting period. A major paper on the use of fungal extracts to reduce honey bee viruses in both cage and fieldcolonies will be published.

Impacts
What was accomplished under these goals? Honey bee breeder queens derived from imported honey bee semen were produced and supplied to western honey bee queen producers Evaluation of colony longevity and health parameters from the "staionary apiary" experiment continued in 2017-2018. Analysis of field colonies indicated that fungal extracts reduced titers of viruses assoicated with parasitic bee mitesin full-sized colonies Indoor wintering protocols were expanded to evaulate the use of metabolic gas and cold storage to reduce brood levels in bees placed in storage during the summer

Publications

• Type: Journal Articles Status: Published Year Published: 2017 Citation: Boyle, N., & Sheppard, W. 2017. A scientific note on seasonal levels of pesticide residues in honey bee worker tissue. Apidologie, 48(1): 128-130.
• Type: Journal Articles Status: Published Year Published: 2017 Citation: Koch, J., Looney, C., Sheppard, W., & Strange, J.P. 2017. Patterns of population genetic diversity and structure across bumble bee communities in the Pacific Northwest. Conservation Genet 18:507-520.
• Type: Journal Articles Status: Published Year Published: 2017 Citation: Gul, A., Sahinler, N., Onal A., Hopkins, B., Sheppard, W. 2017. Effects of diluents and plasma on honey bee (Apis mellifera L.) drone frozen-thawed semen fertility. Theriogenology. 101:109-133.
• Type: Journal Articles Status: Published Year Published: 2017 Citation: Hopkins, B.K., Cobey S. W., & and W. S. Sheppard. 2017. Gel-coated tubes extend above freezing storage of honey bee (Apis mellifera) semen to 439 days with production of fertilized offspring. Reprod. Fertil. Dev. 29:19441949.

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

Outputs
Target Audience: Nothing Reported Changes/Problems: Nothing Reported What opportunities for training and professional development has the project provided?Two postdoctoral researchers are involved in the fungal research at WSUand are receiving training within this project. How have the results been disseminated to communities of interest?Research results from both the fungal research and the indoor wintering/storage experiments were presented to numerous local and regional beekeeping groups (WA, OR, ID, CA, ND, MT) and at national venues, including the American Beekeeping Federation annual convention. What do you plan to do during the next reporting period to accomplish the goals?The experimental stationary apiaries and evaluation of fungal anti-mite and anti-viral outcomes willbe followed through 2018 and the results will be reported in a peer-reviewed journal and within trade journals to the beekeeping industry. We willcontinue research on indoor storage and indoor overwintering of honey bee colonies to assist in control of Varroa mites and reduce overwintering losses.

Impacts
What was accomplished under these goals? An experimental manipulation of metabolic gases using 200 colonies placed in controlled atmosphere storage (Yakima, WA) in 2017 demonstrated that a mid-summer brood break could be accomplished with 18 days of storage in dark conditions at 4C. This breakthen permits the use of "soft" chemical control agents for Varroa mites (i.e. Hopguard) prior to preparation for winter. A "stationary apiary" experiment with 48 full-sized colonies was set up in 2017 to evaluate both an anti-Varroa fungal pathogen (Metarhyzium) and 2 difference fungal extracts to reduce virus titers in field colonies. Previous work in our lab had shown that the extracts reduced viral levels in bees in cages and in small nucleus field colonies.

Publications

• Type: Journal Articles Status: Published Year Published: 2016 Citation: Miguel, I., Garnery, L., Iriondo, M., Baylac, M., Carmen, M., Sheppard, W., & Estonba, A. 2016. Origin, evolution and conservation of the honey bee from La Palma Island (Canary Islands): molecular and morphological data. Journal of Apicultural Research. 54(5).

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

Outputs
Target Audience:Beekeepers, Honey Bee Queen producers, Growers, General Public. Changes/Problems: Nothing Reported What opportunities for training and professional development has the project provided?We conducted workshops on beekeeping methodology, queen production methods and selection of honey bees for PNW conditions. How have the results been disseminated to communities of interest?Through workshops, publications and participation in local, state, regional and national meetings of stakeholders. What do you plan to do during the next reporting period to accomplish the goals?Continue the selection and maintenance of novel honey bee germplasm through the use of instrumental insemination and cryopreservation of honey bee semen. Conduct international collection o honey bee semen from Apis mellifera carnica from Slovenia in 2017.

Impacts
What was accomplished under these goals? Honey bees of three distinct subspecies were maintained and selected germplasm was made available to western US queen producers. Assessment of genetic impact based on micro satellite analysis indicated that significant improvements in honey bee genetic diversity occurred in the populations of honey bees derived from these accessions. WSU was a lead participant in the establishment of a honey bee species committee for the USDA- National Agricultural Germplasm Program.

Publications

• Type: Journal Articles Status: Published Year Published: 2016 Citation: Hopkins B., S. Cobey, C. Herr, W. Sheppard. 2016. Gel-coated tubes extend above-freezing storage of honey bee (Apis mellifera) semen to 439 days with production of fertilised offspring. Reproduction, Fertility, and Development.
• Type: Journal Articles Status: Published Year Published: 2016 Citation: Miguel I., L. Garnery, M. Iriondo, m. baylac, m. carmen, W. Sheppard, A. Estonba. 2016. Origin,evolution and conservation of the honey bee from La Palma Island (Canary Islands): molecular and morphological data. Journal of Apicultural Research.54(5).

Progress 08/11/15 to 09/30/15

Outputs
Target Audience:The target audience includes growers of pollinated crops, beekeepers involved in providing pollination services for agriculture and in producing queens to supply the pollination industry. Changes/Problems: Nothing Reported What opportunities for training and professional development has the project provided?A visiting PhD student from Brasil conducted research on protein gene expression in relation to subspecies variation. A Borlaugh Fellow from Algeria conducted a project on honey bee genetics and reproductive technologies. How have the results been disseminated to communities of interest?During the period of the report, presentations at scientifi and user group meetings were conducted including Beekeeping Associations (CA, OR, ID, WA), grower groups (WA Tree Fruit Research Association)and professional societies (Entomological Society of America) What do you plan to do during the next reporting period to accomplish the goals?Ongoing experiments are underway related to honey bees genetics and breeding (WA and CA), parasite and pathogen control (WA, CA), overwintering (WA), entomopathogenic fungi. Our breeding program continues and genetic material will be distributed to major queen producers in the US. We will collect additional honey bee germplasm from two Old World subspecies in 2016.

Impacts
What was accomplished under these goals? Honey bee germplasm from anOld World subspecies that co-evolved with appleswas collected from Kazahkstan (summer 2015) and used to produce inseminated queens in the US. Portions of the collected semen werecryopreserved and placed into the WSUHoney Bee Germplasm Repository. The inseminated queens are being overwintered and will be used for breeding and selection of improved honey bee strains. previously collected genetic material was distributed to US queen producers. Experimental tests of novel mite and virus controlproducts derived from fungus were conducted and results were analyzed Experiments totestmetabolic gas manipulation as a possible parasite control methods were conducted and results were analyzed

Publications

• Type: Book Chapters Status: Published Year Published: 2015 Citation: Sheppard, W. S. 2015. Honey Bee Diversity - Races, Ecotypes, Strains in The Hive and the Honey Bee. Publ. Dadant and Sons, pp 53-70, 1057 pp.