2008 Annual Report 1a.Objectives (from AD-416) Breed, evaluate and develop Russian honey bees having useful economic traits. Advance the understandings of the mechanisms and associated genetic variance in Russian and other honey bees which underlie useful economic traits in honey bees, such as resistance to Varroa destructor and to Acarapis woodi, honey production, and survival in harsh climates, with the goal of developing improved methods to select stocks of honey bees. Advance the understanding of genetic variance which underlies the variance in honey bee pollination abilities with the goal of producing honey bee stocks improved in their value as pollinators. Develop marker-assisted breeding and stock identification tools and exploit the potential of the sequenced honey bee genome. 1b.Approach (from AD-416) Using a unit designed breeding procedure we will continue to breed Russian honey bee stock for increased mite resistance, honey production, and winter survival. Russian and other stock will be studied to identify specific mechanisms of resistance to parasitic mites and small hive beetle and to find traits that are associated with enhanced pollinator quality. Using Quantitative Trait Loci analysis in association with knowledge provided by the sequencing of the honey bee genome, genetic molecular analysis will be used to identify specific genes having important economic impact on honey bee phenotype, strategies to employ these genes in marker assisted breeding will be developed 3.Progress Report 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 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 with l 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. NP305; Comp 2; PS A.1,A.2,A.3. 4.Accomplishments 1. Full complement of Russian bee breeder lines released to the beekeeping industry. The need exists to provide a final transfer of all Russian bee breeder lines to industry. This goal was accomplished by working with the CRADA holder and a newly formed Russian Bee Breeders Association. Members were systematically provided with breeder queens for queen lines and for drone sources so that each member could produce two of the eighteen lines. In future, they will share stock to develop new drone source colonies and continue to be able to develop all of the lines. This will permit the continued breeding and selection of Russian bees and provide mite resistant Russian honey bee stock to the entire beekeeping industry for the foreseeable future. NP 305, Component 2, Problem Statement: A.1. 2. Low growth of varroa populations in Russian bee colonies is regulated by a variety of mechanisms. Although it was know that Russian honey bees resist varroa mites, the full array of mechanisms producing this resistance was not known. The knowledge of the array of resistance mechanisms is now known. This knowledge permits more targeted selection for resistance to varroa in Russian and other stock. NP 305, Component 2, Problem Statement: A.1. 3. Russian honey bee resistance to tracheal mites results from autogrooming. The mechanism of honey bee resistance to tracheal mites was uncertain. This work confirms that the primary mechanism of resistance is autogrooming by very young adult bees. This advance makes in possible to directly phenotype breeding stock and potentially develop marker assisted breeding. NP 305, Component 2, Problem Statement: A.1. 4. Russian and varroa sensitive hygienic honey bees selectively uncap varroa infested brood cells. It was not know if Russian honey bees used selective hygienic behavior as a mechanism of resistance to varroa. Now it is clear that selective hygiene is part of the suite of characters that impart varroa resistance. Once methods of direct phenotypic measurement are developed there will be a potential to develop marker assisted breeding for the trait. NP 305, Component 2, Problem Statement: A.1. 5. Russian honey bee resistance to varroa mites is enhanced in apiaries having only Russian colonies. The influence of mixing Russian colonies with varroa infested non-resistant colonies in the same apiary was unknown. Data showed that Russian colonies in co-mingled apiaries where under more mite infestation pressure than Russian colonies in Russian only apiaries. This result is important guidance for beekeepers who want to maximize the benefit of having mite resistant Russian honey bees. NP 305, Component 2, Problem Statement: A.1. 6. Honey bees collect large amounts of upland cotton pollen in Louisiana. In dry areas honey bees do not collect upland pollen but such pollen collection was not studied in more humid areas. The discovery that honey bees did collect upland pollen opens the possibility of fostering the selection of hybrid cotton using honey bees as pollinators. NP 305, Component 2, Problem Statement: A.2. 7. Methods developed for successfully requeening Italian colonies with Russian honey bees. Worries have been expressed that it is difficult to introduce Russian queens to Italian colonies. Studies indicated that Russian and Italian colonies accepted both mated Russian queens and Russian queen cells equally well, that requeening colonies with laying workers previously thought to be not able to be requeened and that higher but not economically damaging levels of varroa infestation reduce the rate of requeening success. This array of information allows beekeepers to produce colonies with desired queen stock and increase their rate of success in managing commercial apiaries. These tools are being employed by many beekeepers throughout the country. NP 305, Component 2, Problem Statement: A.1. 8. Both Russian and Italian colonies detect and remove brood infested with eggs and larvae of the Small Hive Beetle (SHB). While African honey bees are noted for hygiene related to SHB, it was not known if European honey bees had this trait. Studies showed that European honey bees have the trait to a high degree although there is variance between colonies. This variance could perhaps be used to select honey bees having increased expression of the trait. NP 305, Component 2, Problem Statement: A.1. 9. Russian colonies had comparatively fewer Small Hive Beetle than Italian colonies. The comparative resistance of Russian colonies to SHB was unknown. Studies showed that Russian colonies carried fewer SHB adults in their colonies especially if entrance reducers were used in hives. This study provides a specific management tool for beekeepers to help control SHB. NP 305, Component 2, Problem Statement: A.1. 10. Comb built by Russian bees interacts with Russian brood in the colony to further reduces the number of varroa mites. The full array of mechanisms of Russian honey bee resistance to varroa mites was not known. The discovery that the greatest reduction in mites in brood in Russian colonies resulted in comb with Russian brood in comb produced by Russian honey bees provides guidance to beekeepers concerning how to maximize the level of resistance to varroa in the Russian colonies that they manage. NP 305, Component 2, Problem Statement: A.1. 11. Washing honey bees in soapy water results in efficient evaluations of varroa mite infestations. To determine mite populations scientists and beekeepers washed bees in ethanol or isopropyl alcohol. In direct comparison, soapy water was just as effective as the more expensive, toxic and polluting alcohols. Many scientists and beekeepers are now routinely using soapy water to evaluate varroa mite populations in their colonies. NP 305, Component 2, Problem Statement: A.1. 12. Genetic diversity of Italian bees in the United States is similar to the diversity of Italian bees in Italy. There was a concern that importation and breeding bottlenecks had endangered the United States stocks to inbreeding. A full survey showed that the diversity of Italian honey bees in the United States was quite high. This provided guidance that there was no need to attempt to enrich the genetic diversity of Italian stocks by importations from Italy. NP 305, Component 2, Problem Statement: A.3. 13. Worker flight activity of Russian and Italian honey bees is equal during both blueberry and almond pollination. Potential differences between the ability of the two stock to pollinate during cooler weather were hypothesized. It was found that both Russian and Italian stocks were equally useful for commercial pollination in the same weather conditions. This result is instrumental for the adaptation of Russian honey bees for commercial pollination. NP 305, Component 2, Problem Statement: A.2, A.3. 5.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. 6.Technology Transfer Number of New Germplasm Releases 1 Number of Web Sites Managed 1 Number of Other Technology Transfer 2 Review Publications Cargel, R.A., Rinderer, T.E. 2007. Acceptance of mated Queens and Queen Cells in Colonies of Russian and Italian Honey Bees. Bee Culture 135(6):27-29 Insuan, S., Deowanish, S., Klinbunga, S., Sylvester, H.A., Wongsiri, S. 2007. Genetic differeniation of the giant honey bee (apis dorsata) in thailand analysed by mitochondrial genes and microsatellites. Biochemical Genetics 45(3/4):345-361 Villa, J.D. Influence of Worker Age on the Infestation of Resistant and Susceptible Honey Bees (Apis mellifera) with Tracheal Mites (Acarapis woodi)*. Apidologie 38:573-578 De Guzman, L.I., Rinderer, T.E., Frake, A.M. 2008. Comparative reproduction of Varroa destructor in different types of Russian and Italian honey bee combs. Experimental and Applied Acarology 44:227-238 Zhou, Y., Kuster, H., Pettis, J.S., Danka, R.G., Gleason, J.M., Greenfield, M.D. 2008. Reaction Norm Variants for Male Calling Song in Natural Populations of Achroia Grisella (Lepidoptera: Pyralidae): towards a Resolution of the Lek Paradox. Evolution 1317-1334.