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?
The added value of honey bee pollination of agricultural crops is estimated at nearly $15 billion annually. The movement and transport of managed bees for commercial pollination purposes, however, poses several important problems and risks to colonies. Honey bee colonies are threatened directly by numerous parasites, pathogens, and pests, including Varroa and tracheal mites, bacterial diseases like American foulbrood (AFB) and an assortment of bee viruses, all of which affect colony well being. Using a combination of chemical, cultural and genetic methods, the Bee Research Laboratory is investigating these threats and developing integrated pest management strategies to deal with them in a safe and environmentally acceptable manner. This project has three specific goals over the next five years:.
1)Investigate honey bee pathogens and mitigate their impact on bee colonies;.
2)improve honey bee colony health through the management of parasitic mites; and.
3)identify and investigate factors that negatively impact queen longevity and durability and develop strategies to mitigate their effect. This research directly contributes to the accomplishment of ARS National Program 305.

2.List by year the currently approved milestones (indicators of research progress)
Year 1 (FY2004) Conduct in vitro laboratory screening of new antibiotics for their ability to inhibit the bacterium that causes American foulbrood disease of honey bees.

Develop analytical assay methods for selected antibiotics.

Modernize our bee disease diagnosis record keeping.

Provide expert diagnosis of honey bee diseases to our customers/stakeholders.

Year 2 (FY2005) Investigate the seasonal incidence of honey bee viruses at the colony level.

Determine the toxicity of selected antibiotics when applied to honey bee colonies and the ability of specific antibiotics to control American foulbrood disease.

Screen potential compounds for their ability to inhibit and control the parasitic protozoan Nosema.

Characterize genomic libraries of honey bees to identify genes involved in the immune response.

Optimize the use of formic acid gel to control parasitic mites of honey bees.

Year 3 (FY2006) Develop an alternative assay for assessing antiprotozoal compounds that do not rely on live honey bees.

Screen the honey bee genome using microarrays for genes involved in the response to disease.

Determine the effects of miticide residues on queen bee performance. Year 4 (FY2007)

Determine the role of the parasitic mite Varroa in the transmission of honey bee viruses.

Conduct and evaluate breeding experiments to verify the contribution of specific genes to their ability to help bees fight disease.

Evaluate drone (male) bee trapping techniques as a form of mite control.

Conduct etiological studies on the effect of Nosema on queen bees.

Year 5 (FY2008) Develop a honey bee cell line that can be used for virus propagation. Collect and submit all necessary data required for FDA registration of antibiotics.

Evaluate field colonies for genetic variation in their responses to disease.

Test and evaluate combined control strategies (formic acid gel, drone trapping, resistant queens) to control parasitic mites of honey bees.

Assess the effect of miticides and Nosema on queen performance and investigate Nosema-induced queen supersedure (replacement) studies.

4a.List the single most significant research accomplishment during FY 2006.
The approval of Tylosin as a new antibiotic for use in honey bee colonies. The approval of Tylosin as a new antibiotic for use in honey bee colonies was granted by FDA in FY 2006 and a use label approved. Data on laboratory research was accepted by the Food and Drug Administration (FDA) and published in the Federal Register and used to support of a New Animal Drug Application (NADA) for the use of tylosin tartrate to control American foulbrood disease of honey bees. This accomplishment addresses the problem that the bacterium that causes this disease has shown widespread resistance to the only antibiotic currently approved for its control. Scientists at the Bee Research Laboratory in Beltsville Maryland, with cooperation from scientists in Weslaco, Texas conducted research on target animal safety, effectiveness, and human food safety. The NADA approval of tylosin for foulbrood control is now providing beekeepers across the U.S. with a new antibiotic to manage this devastating bacterial disease of bees and contribute to maintaining the vitality of the U.S. beekeeping industry. This accomplishment is in direct support of NP 305, component III, development of pest management strategies.

4b.List other significant research accomplishment(s), if any.
Identification of several viruses in queen honey bees. The prevalence of six different bee viruses were determined in queens from several geographic areas and location of these viruses with the queens body were examined. This accomplishment addresses the lack of information surrounding honey bee viral infections in queens. Molecular methods were developed by scientists at the Bee Research Laboratory in Beltsville, Maryland that allowed for the simultaneous detection of six viruses in queen bees that appeared normal. The question of impact of viruses on queens is being investigated. This research is part of a larger effort to improve queen health and increase longevity. This accomplishment is in support of NP 305, component III, development of pest management strategies.

Development of detection methods for tylosin in pollen. A scientist at the Bee Research Laboratory in Beltsville, Maryland developed purification schemes and optimized analytical methods for the detection of the antibiotic tylosin in pollen. The accomplishment addresses the need for accurate detection methods of an antibiotic that will have widespread use in honey bee colonies and can contaminate pollen collected from treated hives. The results of this research will benefit research and commercial operations interested in monitoring levels of this antibiotic in honey. This accomplishment is in support of NP 305, component III, development of pest management strategies.

Honey bee disease genomics. A scientist at the Bee Research Laboratory used genomic approaches to investigate the honey bee immune response. This research addresses the need for understanding the underlying basis for bee responses to disease agents. The scientist showed that non-pathogenic bacteria can induce an immune response in honey bees that may be helpful in warding off infection by a disease-causing bacterium. The scientist also serves on the Advisory Committee for the Honey Bee Genome Project and is coordinating efforts to screen this genome for genes important to honey bee health. These efforts are expected to improve honey bee management and reduce our reliance on commercial antibiotics. This accomplishment is in support of NP 305, component III, bee management and pollination strategies.

4c.List significant activities that support special target populations.
None.

4d.Progress report.
None.

5.Describe the major accomplishments to date and their predicted or actual impact.
Research was conducted to reduce the impact of American foulbrood disease, a devastating bacterial infection of honey bees. New classes of antibiotics were examined for target animal safety, efficacy in controlling the disease, and residues in honey. Reports were submitted to FDA for approval. Tylosin is now approved in the U.S. for use in honey bee colonies to control American foulbrood. This accomplishment is in direct support of NP 305, Component III, the integrated control of bee diseases. Research was conducted to determine if chemicals used to control parasitic mites of honey bees negatively impacted development of honey bee queens. Scientists at the Bee Research Laboratory in Beltsville, MD reared honey bee queens in beeswax containing known amounts of a pesticide used to control parasitic mites. Scientists determined the specific level of pesticide that would inhibit queen development. Results indicated that careful monitoring of pesticide in beeswax is necessary to produce viable queen bees.

Research was conducted to evaluate the role of parasitic mites in transmitting honey bee viruses. Scientists at the Bee Research Laboratory in Beltsville, MD demonstrated conclusively that parasitic mites of honey bees are capable of transmitting viruses from infected bees to uninfected bees. Using molecular techniques, researchers were able to calculate the transmission efficiency from mites to bees, and demonstrate that non-infected mites can acquire virus by sharing the same cell with one or more infected mites. The results of this research provide us with a better understanding of how bee viruses are spread between bees and emphasizes the importance of mite control.

Scientists at the Bee Research Laboratory in Beltsville, MD detected deformed wing virus in honey bees and in parasitic mites of honey bees. This virus had never been detected in the U.S. and may provide an answer to previously unexplained honey bee colony losses. Using molecular techniques, scientists detected this virus in different stages of honey bees, including adult bees with deformed wings, normal-appearing adult bees, eggs, larvae, and pupae. The detection of virus in normal-appearing adults suggests that virus titers may have to reach certain levels before the infection results in deformed wings and has a subsequent negative economic impact. An additional outcome is that the detection of virus in bee stages not associated with mite parasitism suggests there are alternate ways for the virus to spread within colonies.

Scientists at the Bee Research Laboratory in Beltsville. MD developed a laboratory method that allows the simultaneous detection of multiple bee viruses in a single honey bee. Honey bees can often suffer from multiple virus infections and it is not possible to diagnose these infections in the field since many of the viruses produce no apparent symptoms. Using molecular techniques, scientists designed, developed and tested an accurate procedure for detecting six honey bee viruses in a single reaction. This method is quick, accurate and cost-effective, and can be used to isolate and quarantine infected colonies to prevent the spread of disease. The method can also be used by regulatory agencies to determine the virus status of bee colonies from countries interested in exporting bees to the U.S.

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?
A commercial product (tylosin) is now available to beekeepers nationwide to combat American foulbrood. Research on the antibiotic tylosin for the control of American foulbrood disease of honey bees was transferred to the FDA and subsequently made available as Public Master File 5783 as announced in the Federal Register. Scientists have worked closely with Elanco Animal Health (a division of Eli Lilly & Co.) to gain an approved label for a New Animal Drug Application (NADA); this was successful in 2006.

Sequence and expression data of honey bee genes have been registered in public access databases available to other scientists. The entire sequence of the honey bee genome and the bacterium that causes foulbrood disease will be available in FY2006.

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).
Invited presentations of laboratory research to the American Honey Producers Association and the American Beekeeping Federation, January 2006.

Invited presentations on Varroa mite sampling and American foulbrood detection to the Office of International Epizootics (OIE) diseases in Dublin Ireland, August 2005.

Review Publications
Evans, J.D., Armstrong, T.N. 2005. Selective screening for honey bee bacterial symbionts that inhibit a key bacterial pathogen, paenibacillus larvae. Journal of Apicultural Research. 44:168-171.