2006 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? Why does it matter? Varroa destructor, an external parasitic mite of the honey bee, is the most significant 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 nearly 20 years ago. Mites spread rapidly between colonies, and infested colonies historically died. 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. Mites develop resistance to chemicals quickly, and few chemicals are registered and available for mite control. Some beekeeping areas have mite populations that are resistant to the acaricides commonly used for mite control. 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 significant numbers of colonies. A third colony pest, the small hive beetle (SHB), Aethina tumida, has killed many colonies since its discovery in 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, populations of feral honey bees have declined dramatically because of mite parasitism. 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. Honey bee queens imported from Russia through quarantine were used to create colonies which were tested individually, and the best of these 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 several crops. 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 by year the currently approved milestones (indicators of research progress) 2003 a. Test propagate and release Russian breeder lines b. Develop methods of Small Hive Beetle population growth c. Evaluate non-reproduction, phoresy and larval attractiveness as mechanisms of resistance to varroa d. Evaluate autogrooming as a mechanism of resistance in Russian honey bees to tracheal mites e. Initiate development of lines to determine tracheal mite resistance patterns f. Determine temperature and humidity effects on varroa population growth g. Evaluate Russian honey bees as pollinators of cotton h. Use microsatellites to develop an identification system for Russian honey bees i. Develop phenotypic testing procedures to use in a QTL study of an economically important trait 2004 a. Test propagate and release Russian breeder lines b. Initiate a reduced acaricide test with Russian honey bees c. Evaluate honey bee responses to immature stages of Small Hive Beetle d. Continue evaluation non-reproduction, phoresy and larval attractiveness as mechanisms of resistance to varroa e. Evaluate tracheal mite reproduction in Russian honey bees f. Continue development of lines to determine tracheal mite resistance patterns g. Develop choice experiments as a simple method for measuring varroa mite population grow h. Continue determinations of temperature and humidity effects on varroa population growth i. Evaluate Russian honey bees as pollinators of blueberry, cranberry and almonds j. Use an identification system for Russian honey bees to determine feral bee types k. Use phenotypic testing procedures to identify parents for use in a QTL study of an economically important trait 2005 a. Test propagate and release Russian breeder lines b. Continue a reduced acaricide test with Russian honey bees c. Evaluate genetic variance in a comparatively resistant stock honey bee in response to Small Hive Beetle d. Evaluate grooming behavior as a mechanism of resistance to varroa e. Evaluate repellence/attractiveness as mechanisms of resistance to tracheal mites f. Further develop lines to determine tracheal mite resistance patterns g. Evaluate mite counts on adult bees as a simple method for measuring varroa mite population grow h. Screen stocks for cool weather flight i. Use an identification system for Russian honey bees to determine changes in feral bee types j. Collect and phenotype backcross progeny for use in a QTL study 2006 a Test propagate and release Russian breeder lines b. Continue a reduced acaricide test with Russian honey bees c. Further valuate genetic variance in a comparatively resistant stock honey bee in response to Small Hive Beetle d. Evaluate hygienic behavior as a mechanism of resistance to varroa e. Conduct bioassays on extracts of resistant bees to identify mechanisms of resistance to tracheal mites f. Further develop lines and conduct backcrosses to determine tracheal mite resistance patterns g. Evaluate trapping of dead mites as a simple method for measuring varroa mite population grow h. Continue to screen stocks for cool weather flight i. Continue to use an identification system for Russian honey bees to determine changes in feral bee types j. Begin to genotype F1 and backcross progeny for use in a QTL study 2007 a. Test propagate and release Russian breeder lines b. Continue a reduced acaricide test with Russian honey bees c. Determine feasability of breeding bees resistant to Small Hive Beetle d. Evaluate GSI information as a predictor of resistance to varroa e. Further conduct bioassays on extracts of resistant bees to identify mechanisms of resistance to tracheal mites f. Test backcrosses to determine tracheal mite resistance patterns g. Select for cool weather flight h. Continue to use an identification system for Russian honey bees to determine changes in feral bee types i. Finish genotyping F1 and backcross progeny for use in a QTL study 2008 a Test propagate and release Russian breeder lines b. Continue a reduced acaricide test with Russian honey bees c. Conclude determination of feasability of breeding bees resistant to Small Hive Beetle d. Conclude evaluation of GSI information as a predictor of resistance to varroa e. Determine if cuticular chemistry is useful in identifying mechanisms of resistance to tracheal mites f. Conclude testing backcrosses to determine tracheal mite resistance patterns g. Continue to select for cool weather flight h. Continue to use an identification system for Russian honey bees to determine changes in feral bee types i. Evaluate genotypic and phenotypic data for use in a QTL study 4a.List the single most significant research accomplishment during FY 2006. Full complement of Russian bee breeder lines released to the beekeeping industry: The program target of 18 breeder queen lines now has been produced after a decade of testing to identify useful lines. This closed population is a genetically diverse group of bees that are resistant to varroa and tracheal mites, have good honey production, and have been screened against problems such as excessive defensiveness and susceptibility to chalkbrood disease. The availability of a full complement of lines means that the stock has reduced risk of detrimental inbreeding. Program emphasis now has shifted to intensified selection within the breeder lines for improved apicultural traits with a continued emphasis on the release of the best stock from each year's test. 4b.List other significant research accomplishment(s), if any. Low growth of varroa populations in Russian bee colonies is regulated by a variety of mechanisms: Three seasons of evaluation of population growth of varroa mites found growth rates in Russian colonies that consistently were manifold lower than those in Italian colonies. This slower growth rate was supported by lower percentages of infested worker and drone brood cells, lower frequency of brood cells infested with multiple mites, lower mite reproduction and an extended phoretic period. Russian honey bee resistance to tracheal mites results from autogrooming: Although all honey bees develop less attraction for tracheal mites as they age, Russian honey bees have strong resistance to tracheal mites throughout the period of susceptibility present in other stocks. Controlled experiments using worker bees with glued legs to eliminate auto-grooming indicated that Russian honey bees used auto-grooming as their major mechanism of resistance to tracheal mites. 4c.List significant activities that support special target populations. Almost every beekeeper in the United States and many crop growers that benefit from pollination by honey bees are members of target populations. The release of Russian breeding stock, an annual field day for beekeepers and attendance at industry meetings to report results of research that can be directly used in honey bee management. 4d.Progress report. A Specific Cooperative Agreement with Alabama A&M University has the objective of determining the utility of ARS-developed, mite resistant honey bees for beekeepers in Alabama. Russian honey bees and bees with varroa sensitive hygiene (VSH), plus a commercial Alabama bee stock, are being compared for the need for acaricide treatment and for honey production. Colonies were established in Spring 2004 and are being managed by 10 cooperating beekeepers in FY 06. Data to this point show that Russian and suppressed mite reproduction (SMR) stocks performed relatively well in resisting infestation by varroa and tracheal mites and therefore need less frequent treatments with miticides. Queen survival was variable, with Russian queens performing best. This project is ongoing by a no-cost extension through December 2006. A Specific Cooperative Agreement with Louisiana State University is underway to determine the impact of the releases of Russian and SMR honey bees. A survey has been created, beta tested and distributed to the nation's beekeepers. The survey results have been reported in an internal report. Generally both Russian honey bees and SMR honey bees have achieved a degree of industry acceptance and both have room for further acceptance. This constitutes a final report. A Specific Cooperative Agreement with Texas A&M University was established this year to develop a more accessible and informative representation of the honey bee genome. Scaffolds and contigs for chromosomes/groups of the A. mellifera genome were manually joined to form superscaffolds. All sequence features are displayed according to their position on the chromosomes. 5.Describe the major accomplishments to date and their predicted or actual impact. 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 can conduct more practical experiments, and simultaneously beekeepers will benefit commercially. 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 could be underpinned with further research. It is possible that this would foster production of hybrid cotton using honey bees as pollinators. 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, to much more effectively monitor varroa mite populations in their colonies. The accomplishment, "Full complement of Russian bee breeder lines released to the beekeeping industry", is a major milestone for hypothesis 1.1 of the project plan. A status report of the breeding program was delivered in a publication in the major trade journal, and was described at meetings of national and state beekeeping groups. The availability of the entire breeding pool means that the desirable traits of Russian honey bees now can be used with diminished risk of inbreeding problems. In addition to extending the useful life of the stock, this is important for the research program because it allows further selection and improvement of the breeding lines. The accomplishment, "Low growth of varroa populations in Russian bee colonies is regulated by a variety of mechanisms", significantly addresses hypothesis 2.1 of the project plan. Knowledge of the relative importance of mechanisms supporting varroa resistance permits more targeted selection of traits for enhancing resistance. The accomplishment,"Russian honey bee resistance to varroa mites results from autogrooming" sets the stage for direct phenotyping of breeding stock and potentially the development of marker assisted breeding to produce stock resistant to tracheal mites. The accomplishment, "The majority of the current and future research of this project involves refining Russian honey bees as a commercially valuable stock, and studying various traits of the bees to enhance that refinement" is ongoing. The 18 queen lines now commercially available will be genetically selected to retain and further improve several important economic traits: resistance to varroa, resistance to tracheal mites, overwintering ability and honey production. Other research in the project will study aspects such as mechanisms of resistance to parasitic mites, responses to small hive beetles, value in commercial pollination enterprises, their spread into feral populations of honey bees and ways to use honey bee management to best enhance these economically 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 Cooperative Research and Development Agreement (CRADA) with Harper Apiaries. Other ARS scientists in Tucson, AZ, and Weslaco, TX, received experimental queens from Baton Rouge, LA. Although the Russian honey bee breeding program is making stock available to the beekeeping industry, the breeding program is ongoing. Selection within retained breeder lines is now 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 Memorandum of Understanding (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. Project staff made approximately one dozen presentations to state beekeeping associations. 7.List your most important publications in the popular press and presentations to organizations and articles written about your work. (NOTE: List your peer reviewed publications below). Presentations about various aspects of the project were delivered by several staff to the two major national beekeeping organizations (American Honey Producers Association, American Beekeeping Federation) at annual meetings. A laboratory field day was held on Saturday, October 8, 2005, 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. Publication(s): DEGUZMAN, L, FRAKE, A. 2006. Preliminary Results on the Removal Response of Russian Honey Against Brood Infested with Small Hive Beetles. American Bee Journal. 146(5):446. FRAKE, A., DEGUZMAN, L. 2006. Colony Invasion of Small Hive Beetles: The Effects of Honey Bee Type and Entrance Reducers. American Bee Journal. 146(5):447. VILLA, J. 2006. The Effects of Seasonal Fluctuations in Population Densities of Varroa Mites on the Survival of Untreated Colonies. American Bee Journal. 146(5):450. VILLA, J. 2006. Do Tracheal Mites Reduce the Longevity of Workers? American Bee Journal. 146(5):450-451. VILLA, J. 2006. Autogrooming and Bee Age Influence Migration of Tracheal Mites to Russian and Susceptible Worker Honey Bees (Apis Mellifera L). Journal of Apicultural Research and Bee World. 45(2):28-31. DEGUZMAN, L., RINDERER, T., FRAKE, A., TUBBS, H., ELZEN, P., WESTERVELT, D. 2006. Some Observations on the Small Hive Beetle, Aethina tumida Murray in Russian Honey Bee Colonies. American Bee Journal. 146(7):618-620. CARGEL, R., RINDERER, T. 2006. Queen Cell Acceptance in Laying Worker Colonies of Russian and Italian Honey Bees. American Bee Journal. 146(8):698-700. Review Publications Cargel, R.A., Rinderer, T.E. 2006. Queen Cell Acceptance in Laying Worker Colonies of Russian and Italian Honey Bees. American Bee Journal 146(8):698-700. De Guzman, L.I., Rinderer, T.E., Frake, A.M., Tubbs, H., Elzen, P.J., Westervelt, D. 2006. Some observations on the small hive beetle, aethina tumida murray in russian honey bee colonies. American Bee Journal 146(7):618-620. Frake, A.M., De Guzman, L.I. 2006. Colony invasion of small hive beetles: the effects of honey bee type and entrance reducers. American Bee Journal 146(5):447 De Guzman, L.I., Frake, A.M. 2006. Preliminary results on the removal response of russian honey against brood infested with small hive beetles. American Bee Journal 146(5):446 Villa, J.D. 2006. The effects of seasonal fluctuations in population densities of varroa mites on the survival of untreated colonies. American Bee Journal 146(5):450 Villa, J.D. 2006. Do tracheal mites reduce the longevity of workers?. American Bee Journal 146(5):450-451 Villa, J.D. 2006. Autogrooming and bee age influence the migration of tracheal mites to russian and susceptible workers. Journal of Apicultural Research 45(2):28-31