2004 Annual Report 1.What major problem or issue is being resolved and how are you resolving it (summarize project aims and objectives)? How serious is the problem? What does it matter? Varroa destructor, an external parasitic mite of the honey bee, is the number one production problem for the nation's beekeeping industry. Varroa mites have spread to all the mainland states since they were first discovered in North America about 15 years ago. Mites spread rapidly between colonies and colonies die within two years after being infested. Infestation debilitates colonies to the degree that production losses occur in the first year of infestation. Losses include the colonies themselves, lost honey production and pollination, costs associated with the time and labor required to replace colonies, and costs of acaricides and their application to control mites. Mites develop resistance to chemicals quickly, and only two chemicals are registered and available for mite control. Some beekeeping areas have mite populations that are resistant to all of these registered and other unregistered chemicals. A second imported parasitic mite, Acarapis woodi, is the number two production problem for the beekeeping industry. This mite infests the respiratory tracheae of adult honey bees and by feeding and interfering with respiration, debilitates and kills individual worker bees. Cumulative effects are often most intense in honey bee colonies during the winter when they can cause the death of catastrophic numbers of colonies. A third colony pest, the small hive beetle (SHB), Aethina tumida, has killed many colonies since its introduction to the United States in 1998. Larval SHB damage colonies as they feed on bee brood, pollen and honey. Strong colonies can be overwhelmed by high levels of SHB infestation. To reduce SHB infestations, coumaphos and permethrin are used as a colony treatment and soil drench, respectively. However, these chemicals provide only partial control. A fourth problem area is crop pollination. Beekeeping losses because of parasitic mites have resulted in fewer beekeepers and fewer colonies of honey bees nationwide. Concurrently, feral honey bee populations have suffered dramatic losses in colony numbers because of parasitic mites. This has resulted in an increased need for rental colonies to pollinate crops. Changing agricultural practices, including new plant varieties and species, require continual examination of current pollination requirements to assure optimum production of seed, fruit, vegetable, oil and fiber crops. We are developing a stock of honey bees imported from far-eastern Russia which has substantial resistance to the varroa mite and excellent resistance to the tracheal mite. Queen stocks imported from Russia have been brought through quarantine, tested individually and the best are tested in a multi-state field trial. The colonies are evaluated for resistance to each parasitic mite, honey production and overwintering ability. The best lines are included in a breeding program designed to allow further improvement for the important economic traits but also to prevent inbreeding which is highly detrimental to honey bees. Identification of specific resistant traits to both parasitic mites is ongoing so that the genetics of resistance can be studied in detail. Pollination studies are being conducted comparing the pollination effectiveness of Russian and Italian honey bee stocks on lowbush blueberries. Studies include foraging at varied temperatures, colony survival when under moving stress and other characteristics that determine the overall value of a bee stock for commercial pollination. The comparative foraging of the two stocks, as well as the effect of honey bee pollination on cotton quality and quantity, are being compared on upland cotton. 2.List the milestones (indicators of progress) from your Project Plan. A. Test, propagate and release Russian breeder lines according to a schedule organized according to groups of breeder queens. B. Conduct a long-term evaluation of the varroa resistance of Russian honey bees in a reduced acaricide test, and evaluate questions concerning Russian honey bee management. C. Evaluate Russian honey bee mechanisms of resistance to varroa mites. D. Develop simplified measurements of varroa mite population growth. E. Determine varroa mite population growth in contrasting climates. F. Determine whether the basis of Russian honey bee resistance to tracheal mites is auto-grooming. G. Determine the genetics of resistance to tracheal mites in Russian honey bees. H. Compare Russian and Italian colonies to determine potentials for resistance to small hive beetle. I. Determine genetic variation in foraging traits that result in crop pollination that can be exploited or improved in Russian honey bees and develop management techniques for useful improvements. J. Use molecular techniques to identify Russian honey bees and their presence in feral populations of honey bees. 3.Milestones: 3A. List milestones that were scheduled to be addressed in 2004. (a) Test, propagate and release Russian breeder lines according to a schedule organized according to groups of breeder queens. This yearly milestone of testing, propagating and releasing was accomplished. (b) Conduct a long-term evaluation of the varroa resistance of Russian honey bees in a reduced acaricide test and evaluate questions concerning Russian honey bee management. The experimental apiaries for this evaluation were established. Additionally, Russian honey bees were found to best exhibit resistance to varroa in apiaries comprised of only Russian honey bee colonies, meeting this year's milestone. (c) Evaluate Russian honey bee mechanisms of resistance to varroa mites. Russian honey bees have been found to suppress the reproduction of varroa mites and to have heightened levels of phoresy but not to have depressed larval attractiveness to varroa, meeting this and next year's milestones. (d) Develop simplified measurements of varroa mite population growth. Tests of mechanically shaking containers with soapy water to separate varroa mites from samples of adult bees showed that the procedure was faster, less expensive, more effective and did not produce waste disposal concerns in comparison to the commonly used technique of hand washing with ethyl alcohol. Tests of different techniques of conducting choice experiments where mites chose between brood types were conducted showing that brood attractiveness was not a strong mechanism of resistance or that it could be used as a selection assay. This work met this year's milestone and future work will concentrate on other possible simplified measurements. (e) Determine varroa mite population growth in contrasting climates. Simultaneous measurements of humidity and temperature in brood areas of colonies in Louisiana and Arizona showed no significant differences. As indicated in the Project Plan contingency, no further research was planned if these measurements showed no differences. Milestone accomplished. (f) Determine whether the basis of Russian honey bee resistance to tracheal mites is auto-grooming. It has been found that Russian honey bees do auto-groom tracheal mites and that this behavior is an important aspect of their resistance to tracheal mites. Russian and susceptible honey bees exhibited differential responses to auto-grooming impairment, suggesting that bees have additional mechanisms of resistance to tracheal mites. Milestone met. (g) Determine the genetics of resistance to tracheal mites in Russian honey bees. Two queen lines of both highly resistant Ruissian and susceptible domestic bees were selected and parents will be screened this fall to select which ones to use to produce F1 colonies. Milestone has been met. (h) Compare Russian and Italian colonies to determine potentials for resistance to small hive beetle. In a two-apiary experiment, Russian honey bees survived longer with fewer hive beetles suggesting they have some level of resistance to SHB. Experimental colonies for a second experiment have been established, meeting this year's milestone. (i) Determine genetic variation in foraging traits that result in crop pollination that can be exploited or improved in Russian honey bees and develop management techniques for useful improvements. Foraging performance of Russian honey bees was studied under marginal weather conditions during pollination of lowbush blueberries in Maine. A preliminary test of feeding protein to stimulate early-season growth of Russian and Italian colonies did not indicate a benefit for either stock. The experiment suggested possible directions for future studies. Planning continues for a possible importation of Apis mellifera pomonella, a sub-species from central Asia that is adapted to an area where apple trees evolved. A proposal for importation is being prepared for submission to USDA-APHIS and the Louisiana Department of Agriculture and Forestry. Collectively, these activities have met this year's milestones. (j) Use molecular techniques to identify Russian honey bees and their presence in feral populations of honey bees. Sixty-three swarms collected in 2003 were identified as primarily Russian (30%), Hybrid (13%) and non-Russian (57%). Two hundred swarm traps placed in 2004 yielded 120 swarms which will be analyzed molecularly this coming FY, meeting this year's milestone. (k) Conduct QTL analyses of economically valuable traits. Due to staffing limitations this research goal was merged with 3.3 of 6413-21000-008-11D. The recent publication of a high-density microsatellite DNA map obviated the need to find other DNA markers for this study. Based on field tests, the characteristic of chalkbrood resistance was identified as the most suitable economically valuable trait of several traits studied for QTL identification, replacing pollen foraging QTL's. Susceptible and resistant colonies were identified in field tests and 250 candidate microsatellite DNA markers were chosen, meeting this year's milestone. 3B. Expected accomplishment under each milestone for the next 3 years. (a) Test, propagate and release Russian breeder lines according to a schedule organized according to groups of breeder queens. FY 05, FY 06, FY 07: Each year, test, propagate and release Russian breeder lines. Each year, test one management question about Russian honey bees raised by beekeepers. (b) Conduct a long-term evaluation of the varroa resistance of Russian honey bees in a reduced acaracide test. FY 05, FY 06, FY 07: Continue the long term test until Russian colonies die of varroa (Prior experimentation indicates that the Italian colonies will require replacement because of loss to mites during the course of the experiment). A new test may be started using the latest selected products from the breeding program, perhaps in FY 06 or FY 07, to estimate levels of stock improvement. (c) Evaluate Russian honey bee resistance mechanisms to varroa mites. FY 05: Further evaluation of non-reproduction and phoresey. FY 06: Evaluation of grooming behavior. FY 07: Evaluation of resistance to invasion pressure or GIS analyses of micro-climate effects. (d) Develop simplified measurements of varroa mite population growth. FY 05: Evaluate correlations of components of mite populations with total mite populations. FY 06: Develop mite choice experiments as a potential simplified measurement. FY 07: Evaluate methods to estimate mite counts on adult bees. (e) Determine varroa mite population growth in contrasting climates. Since no environmental effects were found in comparisons of the highly contrasting enviroments of Louisiana and Arizona, this research objective will not be pursued further. (f) Determine whether the basis of Russian honey bee resistance to tracheal mites is auto-grooming. FY 05: Study auto-grooming of tracheal mites by Russian honey bees. FY 06: Study chemical repellence as a mechanism of tracheal mite resistance. FY 07: Conduct bioassays on extracts of resistant bees. (g)Determine the genetics of resistance to tracheal mites in Russian honey bees. FY 05: Evaluate F1 crosses of resistant and susceptible bees. FY 06: Evaluate backcross colonies of F1 to presumed recessive stock. FY 07: Begin QTL analysis of samples preserved from parental, F1 and backcross colonies. (h) Compare Russian and Italian colonies to determine potentials for resistance to small hive beetle. FY 05: Conduct second replication of comparative field trial, study behavioral responses to immature stages of SHB. FY 06: Assess genetic variance in a comparatively resistant population. FY 07: Continue studies of genetic variance. (i) Determine genetic variation in foraging traits that result in crop pollination that can be exploited or improved in Russian honey bees and develop management techniques for useful improvements. FY 05: Continue studies on lowbush blueberry, initiate studies on cranberry and evaluate the colony development response of Russian honey bees to protein and sugar feeding to increase their size prior to almond pollination. FY 06: Screen stocks of honey bees for cool weather flight. FY 07: Begin selection for improvements in cool weather flight. (j) Use molecular techniques to identify Russian honey bees and their presence in feral populations of honey bees. FY 05, FY 06, FY 07: Continue collection and identification of swarms near Russian and non-Russian apiaries evaluating trends. (k) Conduct QTL analyses of economically valuable traits. FY 05: Production of F1 and backcross colonies and field evaluations of backcross colonies. FY 06: Beginning molecular evaluations. FY 07: Finish molecular evaluations and analyze data. 4.What were the most significant accomplishments this past year? 4A. Single most significant accomplishment during FY 2004 (one per Research (OOD) Project): Russian honey bee resistance to varroa mites is enhanced in apiaries having only Russian colonies. Management of varroa resistant stocks to maximize their resistance is as important as having resistant stocks. An experiment compared the effects of co-mingling or not co-mingling resistant Russian and susceptible Italian stocks in both sunny and shaded locations. Results showed that while resistant stock always had fewer mites than susceptible stock, they had far fewer mites when not in the same apiary as susceptible stock which served as a source of infestation for the entire apiary. This result provides important guidance for beekeepers exploring the value of resistant stocks in their individual beekeeping environments. B. Other significant accomplishments, if any. Honey bees collect large amounts of upland cotton pollen in Louisiana. An experiment comparing Russian and Italian honey bee stocks found both stocks collecting large amounts of upland cotton pollen. This result contrasts dramatically from experiments in Arizona where honey bees only very rarely collected cotton pollen. While cotton pollen collection in Louisiana by honey bees may possibly be due to high humidity, the results underpin observations by others that honey bee pollination of upland cotton results in increased yield of lint. If further observations show that honey bees often collect cotton pollen in specific areas of the country and that the collection does indeed increase the value of the cotton crop, cotton farmers and beekeepers in those areas would economically benefit. Washing honey bees in soapy water results in efficient evaluations of varroa mite infestations. Removing varroa mites from samples of honey bees has involved hand washing in ethyl alcohol. This procedure is time consuming, costly and produces a waste product that requires special handling. Tests of mechanically shaking containers with soapy water to separate varroa mites from samples of adult bees showed that the procedure was faster, less expensive, more effective and did not produce waste disposal concerns, in comparison to the commonly used technique of hand washing with ethyl alcohol. This research greatly enhances the ability to quickly and efficiently evaluate mite population levels in honey bee colonies. C. Significant activities that support special target populations. Almost every beekeeper in the United States and many growers that benefit from pollination by honey bees are members of target populations. D. Progress Report opportunity to submit additional programmatic information to your Area Office and NPS (optional for all in-house ("D") projects and the projects listed Appendix A; mandatory for all other subordinate projects). A subordinate project is a cooperative program with Alabama A and M has the objective of determining the utility of ARS-developed, mite resistant honey bees for beekeepers in Alabama. Russian honey bees and SMR honey bees plus a commercial Alabama bee stock are being compared for the need for varroa treatment and for honey production. Colonies were established in Spring 2004 and are being managed by eight cooperating beekeepers. Data are not yet available; the project is being continued in next FY. 5.Describe the major accomplishments over the life of the project, including their predicted or actual impact. This is the first year of this project. Hence, the major accomplishments of this project thus far are reported in section 4 of this year's report. The accomplishment "Russian honey bee resistance to varroa mites is enhanced in apiaries having only Russian colonies" is a major milestone for section 1.1 of the project plan. This accomplishment was transferred to industry by publication in a trade journal and various talks to beekeeping groups. Using this knowledge will allow scientists in this unit and elsewhere and beekeepers to better assess the varroa mite resistance of Russian honey bees and benefit commercially, respectively. The accomplishment "Honey bees collect large amounts of upland cotton pollen in Louisiana" is a major milestone for section 3 of the project plan. This accomplishment is currently being underpinned with further research. When technology transfer takes place, at least some cotton growers and beekeepers renting colonies for pollination will benefit commercially. The accomplishment "Washing honey bees in soapy water results in efficient evaluations of varroa mite infestations" is a major milestone for section 2.3 of the project plan. This accomplishment was transferred via a publication in a trade journal and several talks at beekeeper meetings. Using this technique will allow scientists in this unit and elsewhere and beekeepers that do so, to much more effectively monitor varroa mite populations in their colonies. The large majority of this project's plans focus on the further development of Russian honey bees as a commercially valuable stock and the study of various traits of Russian honey bees to enhance that development. In a prior project, Russian honey bees were discovered to show resistance to varroa in Russia, were imported through quarantine into the United States, evaluated and found to have variable but useful resistance to varroa, excellent resistance to tracheal mites and excellent ability to overwinter in even the extremes of northern locations. Over 170 queen lines were evaluated in single colony or multi-colony sib tests in several states for possible inclusion into a breeding program. This is the last year imported queens will be evaluated and it is anticipated that about 15 queen lines will be retained. Quality queen lines found in prior year have been made available to the beekeeping industry through a CRADA. These queen lines will be genetically selected during the course of this project to further improve or retain several important economic traits: resistance to varroa, resistance to tracheal mites, overwintering ability and honey production. Other research in the project will study various aspects of Russian honey bees such as resistance mechanisms to parasitic mites, small hive beetles, their value in commercial pollination enterprises, their spread into feral populations of honey bees and ways to use honey bee management to best enhance these economially important traits in Russian and other honey bee stocks. 6.What science and/or technologies have been transferred and to whom? When is the science and/or technology likely to become available to the end-user (industry, farmer, other scientists)? What are the constraints, if known, to the adoption and durability of the technology products? Russian honey bee breeder lines were transferred to the beekeeping industry through a CRADA. Other ARS scientists in Tucson and Weslaco received experimental queens from Baton Rouge. Although the Russian honey bee breeding program is making stock available to the beekeeping industry, the breeding program is ongoing. This is the last year that untested queen lines from Russia will be evaluated and chosen for inclusion in to breeding program or culled. In coming years, selection within retained breeder lines will be more intense and is expected to produce a stock with higher and less variable expression of traits. Many producers of "Russian queens" for sale to the public are not mating the queens in places that have drones from Russian colonies. Hence, they are actually selling queens that produce hybrid colonies that likely show high variability and sometimes show undesirable traits such as being sting-prone. These hybrid products have been rejected by some customers, many of whom are seeking sources of "pure bred" Russian stock. Although more queen breeders are developing businesses based on "pure bred" stock, it is clear that the demand currently far exceeds supply. Information about using only resistant stock in apiaries and washing bees with soapy water on a mechanical shaker has been transferred to other scientists and beekeepers by publications in a trade journal and talks at beekeeper meetings. All beekeepers should be able to easily establish apiaries with only one type of stock. Only some beekeepers monitor levels of varroa in their colonies and may have problems developing mechanical shakers. More information about what mite levels can be considered triggers for treatment of resistant stock is needed before most beekeepers would consider using the technique to monitor levels of varroa. Under a MOU, Backwood Apiaries currently offers tests for tracheal mite resistance of commercial queens. The unit provides resistant and susceptible queens to be used as standard references. A laboratory field day was held on Saturday, October 18, 2003, for about 100 beekeepers from the region. Beekeeping and research activities were shown ranging from simple for beginners to complex for commercial operators and specialized hobbyists. Project staff made approximately 19 oral presentations at national and state beekeeper association meetings. 7.List your most important publications in the popular press and presentations to organizations and articles written about your work. Kaplan, Kim J. 2004. What's Buzzing with Africanized Honey Bees? Agricultural Research 4:4-8. Review Publications Rinderer, T.E., De Guzman, L.I., Sylvester, H.A. 2004. Reexamination of the accuracy of a detergent solution for varroa mite detection. American Bee Journal. Vol. 144(7):560-562 Rinderer, T.E., De Guzman, L.I., Harper, C. 2004. The effects of co-mingled russian and italian honey bee stocks and sunny or shaded apiaries on varroa mite population growth, worker bee population and honey production. American Bee Journal. Vol. 144(6):481-485. Gregory, P.G., Rinderer, T.E., Harbo, J.R. 2004. Paternity of offspring from honey bee queens re-inseminated after producing worker brood [abstract]. American Bee Journal. 144(5(:404 Danka, R.G., Villa, J.D. 2004. Contemporary soybeans show no evidence of yield increases associated with proximity to honey bee colonies [abstract].American Bee Journal. 144(5):402 Danka, R.G. 2004. High levels of cotton pollen collection and foraging responses of italian and russian honey bees given high and low levels of pollen foraging stimuli [abstract]. American Bee Journal. 144(5):402 Gregory, P.G., Rinderer, T.E. 2004. Non-destructive dna sampling in honey bee queens, apis mellifera. Entomologia Experimentalis et Applicata. Vol. 111:173-177. Danka, R.G., Villa, J.D. 2003. Autogrooming by resistant honey bees which are challenged by individual tracheal mites.. Apidologie 34(4):591-596. Tubbs, H., Harper, M., Bigalk, M., Bernard, S.J., Delatte, G.T., Sylvester, H.A., Rinderer, T.E. 2003. Commercial management of ARS russian honey bees. American Bee Journal 143(10):819-820. Villa, J.D. 2004. Swarming behavior of honey bees (hymenoptera: apidae)in southeastern Louisiana. Annals of the Entomological Society of America 97(1):111-116. Kavinseksan, B., Wongsiri, S., De Guzman, L.I., Rinderer, T.E. 2004. Absence of tropilaelaps infestation from recent swarms of Apis dorsata in Thailand. Journal of Apicultural Research. 42(3):49-50.