Progress 10/01/07 to 09/30/12
Outputs
OUTPUTS: Candidate compounds for the control of the pathogen Nosema ceranae in honey bees were tested on infected bees in cages. The compounds include a fungicidal protein isolated from tobacco leaves and 2-deoxyglucose, which interferes with glucose metabolism. A technique for inoculation of individual worker bees with known doses of spores was tested and perfected. By controlling the inoculum given to individual bees, the variance in the results from controlled studies may be reduced. This should then allow us to see effects of factors of interests, for example candidate substances for the control of Nosema infection. This is based on an early publication by Rinderer (1976). The progress of Nosema infection in worker honey bees was monitored, by inoculating many individual bees each with 50,000 spores in sucrose solution. The development of immature and mature Nosema forms was timed, and these Nosema forms were characterized according to their morphological features. A method for monitoring temperatures in honey bee hives was developed, so that the effects of Nosema on wintering clusters may be monitored. Two scientific presentations were made: Webster, T. C., E. Hogue and M. Matisoff. "Nosema spore viability is affected by fecal matter" and Matisoff, M. and T. Webster. "Effects of osmotic pressure, ionic composition, sugars, temperature and time on Nosema polar filament composition" at the annual American Bee Research Conference, Feb. 7-8, 2012, Greenbelt MD. A joint workshop including members of the American Association of Professional Apiculturists and the Apiary Inspectors of America was initiated by TW. This allowed for an interchange of ideas and observations, between those who meet with many beekeepers (AIA) and those who conduct research on honey bees (AAPA). PARTICIPANTS: Martin Matisoff, Research Assistant at Kentucky State University, performed much of the research and related duties on this project, under supervision of the P.I., T. Webster. Dr. Kate Aronstein, researcher at USDA-ARS Honey Bee Research Lab in Weslaco TX, collaborated by developing antibodies to Nosema ceranae, and in writing part of the co-authored book chapter. Her laboratory technician, Eduardo Salvidar, also participated in the antibody development. T. Webster participated in the CCD Steering Committee meeting, Oct. 16-17 at Arlington VA. This was an opportunity to guide discussions and research on honey bee mortality, and to learn from others in research, government, commodity groups and industry. Approximately 100 attended. TARGET AUDIENCES: Beekeepers were audiences to discussions and workshops pertaining to the research results from this project, and implications of the research. Presentations and workshops were made at Somerset KY (33 attendees), Frankfort KY (492), Clarkson KY (451), Frankfort KY (37), St. Louis MO (295), Morehead KY (43), and Springfield IL (203). PROJECT MODIFICATIONS: Nothing significant to report during this reporting period.
Impacts
New candidate treatments for Nosema in honey bees did not show promise in early trials, although further studies are planned. The method for inoculating many individual bees was highly successful, after some changes in the method. The appearance of primary spores and environmental spores was described, and corresponding staining techniques. These observations may be used for rapid evaluations of new treatments for Nosema in honey bees, or evaluations of bee stock thought to be resistant to this pathogen. Approximately 100 members of the AIA and AAPA shared observations and research results. This included discussion of increased European foulbrood in southeastern states, difficulties in the identification of honey bees with African bee DNA, and approaches to the sampling of bees with suspected disease.
Publications
- Webster, T. C. and Aronstein. K. 2012. Nosema ceranae detection by microscopy and antibody tests, Chapter 10 In D. Sammataro and J. A. Yoder (eds.), Honey Bee Colony Health: Challenges and Sustainable Solutions. CRC Press. Boca Raton, FL, p. 115-120.
- Webster, T. C., E. Hogue and M. Matisoff. 2012. Nosema spore viability is affected by fecal matter. In Proceedings of the American Bee Research Conference, p. 404 in Amer. Bee J., April 2012.
- Matisoff, M. and T. Webster. 2012. Effects of osmotic pressure, ionic composition, sugars, temperature and time on Nosema polar filament composition. In Proceedings of the American Bee Research Conference, p. 407 in Amer. Bee J., April 2012.
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Progress 01/01/11 to 12/31/11
Outputs
OUTPUTS: As vice president of the American Association of Professional Apiculturists (AAPA) , Webster coordinated presentations by apiculture researchers and logistics for the joint meeting of AAPA, the American Beekeeping Federation and the American Honey Producers, at Galveston TX, Jan 5-7. Webster took the role of the AAPA president at that meeting and guided plans to meet again in 2012, and future roles of AAPA. Webster also presented research results on the bee pathogen Nosema ceranae, at the Galveston event and also at an Invasive Species conference in Lexington, KY May 3-5. Webster organized and presented research to about 500 regional beekeepers at the annual Heartland Apicultural Society conference in Vicennes, IN. Outreach to non-beekeepers were by presentation of research to the Kentucky legislators at Frankfort, Feb 15, and by interview to WKYT television (Lexington) on the value of honey bees. Presentations to beekeepers on Nosema ceranae were at the Spring (April 16, KSU), Summer (June 4, Clarkson) and Fall (Oct 22, Somerset) conferences of the Ky State Beekeeping Assocition. Local associations of beekeepeers were addressed at Muhlenberg (March 21), Scott (May 17) and Franklin (Sept 27) counties. Webster organized the establishment of an observation hive in Nicholas County which was used to teach thousands of children at a 4H camp there. PARTICIPANTS: Numerous partners in research include: Drs. L. Solter (Univ IL), Z. Huang (Mich St Univ), K. Aronstein (USAD_ARS Weslaco TX), J. Ellis (Univ FL), K. Delaplane (Univ GA), W. Hood (Clemson Univ), J. Skinner (Univ TN), G. Hunt (Purdue Univ), M. Spivak (Univ MN), Y. Chen (USDA-ARS Beltsville MD) -- all particpants in the CAP project. See www.beeccdcap.uga.edu Also, Drs G. Wagner (Univ KY), K. Pomper and J. Sedlacek (both KY St Univ) have been valuable collaborators on selected research projects. Four undergraduate and one graduate student participated in research projects at KSU, supervised by Webster. TARGET AUDIENCES: Target audiences are primarily researchers and apiary inspectors, who are concerned with minimizing bee colony mortality, and also beekeepers who are affected by the honey bee pathogens Webster studies. To a lesser extent, target audiences are children and also adults who are not beekeepers. These non-beekeepers should be kept aware of the value of honey bees in the role of crop pollinators. PROJECT MODIFICATIONS: Due to heavy turnover in research and support staff, the project was extended for an additional year. This will allow suitable time for collection of data as planned.
Impacts
Scientific presentations have shown researchers and beekeepers how Nosema ceranae affects honey bee hives. A closer look at the biology of the disease has demonstrated how others might approach this pathogen with greater effect. Research in collaboration with Dr Aronstein at the USDA-ARS laboratory in Weslaco, TX has allowed us to show the feasibility of a new and simple test kit for beekeepers. When this kit is commercialized we may expect a very substantial economic impact for American beekeepers. Numerous discussions by Webster with colleagues who study honey bees have lead to an improved understanding by these colleagues on the biology of Nosema ceranae in honey bee hives. This understanding has lead to more effective research projects by these colleages, which in turn are likely to lead to economic impact by lower bee mortality for American beekeepers.
Publications
- Aronstein, K. A., E. Saldivar, and T. C. Webster. 2011. Evaluation of Nosema ceranae spore-specific polyclonal antibodies. J. Apic. Res. 50(2): 145-151.
- Matisoff, M. and T. C. Webster. 2011. Anatomical changes in the peritrophic membrane associated with Nosema ceranae. Amer. Bee J, May.
- Webster, T. C. and M. Matisoff. 2011. The appearance of primary spores of Nosema ceranae, as an indicator of infection. Amer. Bee J., May.
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Progress 01/01/10 to 12/31/10
Outputs
OUTPUTS: In a collaborative project with Dr. Steve Pernal of Agri-Canada, three treatments were evaluated for their efficacy in killing Nosema ceranae spores on beekeeping equipment. These were an electron beam apparatus, fumigation with 80% acetic acid, and heat treatments at 49C. Trypan blue stain was used to determine whether these agents ruptured spore membranes in the process of destroying spore viability. Bees established on treated equipment were evaluated later for N. ceranae infection. A method to monitor bee hive temperatures was been developed, in order to evaluate honey bee colony temperatures remotely and non-invasively. By this method colonies that are suffering from insufficient food, disease, parasites, and queenlessness may be identified before they die, so that details and remedial actions may be better understood. This method will be employed to determine overall strategies for reducing honey bee colony mortality, particularly in winter when hives cannot be opened without disturbing the bees. Samples taken from hives in Tennessee and Florida were evaluated for N. ceranae by the polymerase chain reaction technique. These data will be combined with data from five other states to establish an understanding of the distribution of this pathogen geographically and according to beekeeping technique. PARTICIPANTS: Dr. Steve Pernal directs reseach at Agri-Canada, in Beaverlodge, Alberta. Dr. Abdullah Ibrahim is a post-doctoral researcher at the same site. Both are collaborators on the study of treatments to kill N. ceranae spores on beekeeping equipment. Dr. Jeff Ashley is a researcher at the Center for Manufacturing at University of Kentucky and Dr. Siddhartha Bhattacharyya is an associate professor in the Computer Science Department at Kentucky State University. Both collaborated on the method for monitoring bee hive temperatures. Drs. Z. Huang (Univ. Michigan), M. Spivak (Univ Minnesota), G. Hunt (Purdue Univ.), J. Skinner (Univ Tennessee), M. Hood (Clemson Univ.) and J. Ellis (Univ Florida) are collaborators on the multi-state study of the distribution of N. ceranae. TARGET AUDIENCES: Target audiences were colleagues who study apiculture and related fields, and beekeepers in Kentucky and nearby states. PROJECT MODIFICATIONS: Nothing significant to report during this reporting period.
Impacts
A study of three types of treatment for the elimination of N. ceranae spores on beekeeping equipment shows that an electron beam treatment is highly effective, while acetic acid fumigation and heat are less effective. Data from seven states indicate the wide distribution of N. ceranae in a north-south transect. Presentations to large meetings of beekeepers included dissemination of study results regarding treatments and distribution of N. ceranae in honey bee hives. These were at locations across Kentucky on March 13 and 27, April 17, June 5 and November 6. Also, presentations to large meetings of beekeepers were made in West Virginia on April 10 and in Tennessee on July 8 - 10.
Publications
- Webster, T.C., F. E. Dowell, E. B. Maghirang, E. M. Thacker. 2010. Does visible/near-infrared spectroscopy detect honey bee characteristics Amer. Bee Journal. 150(5):509.
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Progress 01/01/09 to 12/31/09
Outputs
OUTPUTS: Honey bee samples from Kentucky, Minnesota, Michigan and South Carolina were analyzed for the presence of the pathogens Nosema ceranae and Nosema apis. This was done by examining bees for spores by light microscopy, extracting DNA from the samples with spores, and identifying the Nosema by species using the polymerase chain reaction. Comb inoculated with N. ceranae spores was treated with either irradiation by electron beam, fumigation with acetic acid, heat at 49C, or control (no treatment). The comb was then placed inside bee hives, and the bees were sampled later to determine N. ceranae infection. Webster, T. C. and M. L. Calhoun. Radio interview with Earthworms show, St. Louis Public Radio, February 2009. Colony Collapse Disorder, Powell county beekeepers, February 2009. Beginning Beekeeping, and Nosema Disease, Bluegrass Bee School, March 2009. Colony Collapse Disorder, Corbin area beekeepers, May 2009. Queen Bee Rearing workshop, Kentucky State University, May 2009. Queen Bee Rearing Workshop, Grayson county, June 2009. Colony Collapse Disorder, and Nosema disease workshops; presented to Heartland Apicultural Society annual conference, July 2009. Honey Bees, to Cane Run Elementary School, Louisville, October 2009. Nosema Disease and Nosema Workshop, to Ohio State Beekeeping Assoc, October 2009. Methods for evaluating the viability of Nosema ceranae spores extracted from honey bees (Apis mellifera); presented to the annual conference of the Entomological Society of America, December 2009. Two undergraduates and one high school intern mentored in research techniques pertaining to Nosema disease in honey bees. PARTICIPANTS: Dr. Floyd Dowell and Elizabeth Maghirang, of the USDA-ARS Grain Marketing and Production Research Center at Manhattan KS are researchers expert in the use of visible and infrared spectroscopy to detect qualities of biological material, especially insects. Dr. Kirk Pomper, Dr. Li Lu, Jeremy Lowe and Kyle Schneider are researchers at the Land Grant Program, Kentucky State University and expert in the use of polymerase chain reaction (PCR) for the detection of DNA in honey bee pathogens. Drs. Marla Spivak, Zachary Huang, Greg Hunt, Michael Hood and James Ellis are faculty members of University of Minnesota, Michigan State University, Purdue University, Clemson University and University of Florida, respectively. They sampled bees in their respective states, for the north-south transect designed to show the distribution of Nosema spp. Melissa Calhoun is a Research Assistant at the Land Grant Program, Kentucky State University. TARGET AUDIENCES: Audiences are scientists who study honey bee pathogens, general apiculture, and pathogens of other economically important insects; beekeepers who must control the pathogens Nosema apis and Nosema ceranae; growers of bee-pollinated crops which depend on healthy honey bee colonies. PROJECT MODIFICATIONS: A survey of Kentucky counties for Nosema ceranae and Nosema apis was expanded to include Minnesota, Michigan, Indiana, South Carolina and Florida. Samples from Tennessee are expected also. This change was made to allow a north-south transect across much of the U.S. The additional data will allow a more thorough evaluation of the effects of latitude, and of beekeeping practices, on Nosema infection. Participation by colleagues in those states has made this expanded sampling possible.
Impacts
Bees from 8 of 11 Kentucky counties and 18 of 55 hives were found to contain N. ceranae. Bees from 42 of 58 Minnesota hives, 23 of 43 Michigan hives, and 6 of 50 South Carolina hives were positive for N. ceranae. No samples contained N. apis. Bees sampled from hives that had earlier been exposed to N. ceranae spores on comb, were found to be heavily infected with N. ceranae if the comb had received electron beam treatment. Heat and acetic acid treatments were not highly effective in suppressing infection over a 3-month sampling period. Beekeepers who attended workshops on Nosema disease learned how to prepare samples and observe the disease using a microscope. This will allow them rapid and inexpensive diagnosis for their hives and hives belonging to other nearby beekeepers. Beekeepers who attended presentations on Colony Collapse Disorder learned about the best methods for maintaining hive survival and vigor.
Publications
- Webster, T. C., F. E. Dowell, E. B. Maghirang, and E. M. Thacker. 2009. Visible and near-infrared spectroscopy detects queen honey bee insemination. Apidologie. 40:565-569.
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Progress 01/01/08 to 12/31/08
Outputs
OUTPUTS: Visible and infrared spectra of honey bee semen were obtained and compared to spectra for virgin and mated queen honey bees. The spectra allowed virgin queens to be distinguished from mated queens, but the semen in spermathecae of mated queens contributed little to this difference. Spores of the honey bee pathogens Nosema apis and Nosema ceranae were treated with selected alcohols, hydrogen peroxide, paradichlorobenzene, ApiLifeVAR, heat and ultraviolet light. All had measureable effects. Membranes of N. apis spores were damaged most by ultraviolet light and hydrogen peroxide, while N. ceranae spores were most damaged by heat, ApiLifeVAR and paradichlorobenzene. Webster, T.C. and Thacker, E.M. The effects of selected agents on Nosema apis and Nosema ceranae spore viability. To a joint meeting of American Bee Research Conference, Apiary Inspectors of America, American Beekeeping Federation, and the American Honey Producers Association, January 2008, Sacramento CA. Webster, T.,C., Dowell, F.E., Maghirang, E.B., and Thacker, E.M. Visible and Near-Infrared Spectroscopy Detects Honey Bee Characteristics. To the Annual Conference of the Kentucky Academy of Science. November 2008, Lexington KY. Nosema disease. Colony Collapse Disorder. Kentucky queen rearing and bee breeding project. Fall Conference of the Kentucky State Beekeeping Association, July 2008, Christian Co., KY. Nosema diagnosis and control. Beekeeping pests. Beekeeping diseases. Conference of the Heartland Apicultural Society, Huntington, WV July 2008. Nosema diagnosis and control. Colony collapse disorder. Conference of the Eastern Apicultural Society, Murray, KY, August 2008. One high school intern was mentored in a study of Nosema spores. Colony collapse disorder. National Public Radio, Paducah, KY, August 2008. Wax moths. Breeding bees for disease and mite resistance. Nosema disease. Bell Co. Field Day, Bell Co. KY, August 2008. Wax moths. Nosema disease. Lewis Co. beekeepers association. Vandenburg. KY, August 2008. PARTICIPANTS: Dr. Floyd Dowell and Elizabeth Maghirang, of the USDA-ARS Grain Marketing and Production Research Center at Manhattan KS are researchers expert in the use of visible and infrared spectroscopy to detect qualities of biological material, especially insects. Dr. Kirk Pomper and Jeremy Lowe are researchers at the Land Grant Program, Kentucky State University and expert in the use of polymerase chain reaction (PCR)for the detection of DNA in honey bee pathogens. Dr. Greg Hunt is a researcher at the Department of Entomology, Purdue University, and expert at the use of PCR. Etta Thacker was a Research Assistant at the Land Grant Program, Kentucky State University. Melissa Calhoun is a Research Assistant at the Land Grant Program, Kentucky State University. TARGET AUDIENCES: Audiences are scientists who study honey bee pathogens, general apiculture, and pathogens of other economically important insects; beekeepers who must control the pathogens Nosema apis and Nosema ceranae; growers of bee-pollinated crops which depend on healthy honey bee colonies. PROJECT MODIFICATIONS: Nothing significant to report during this reporting period.
Impacts
Researchers became aware of the significance of selected treatments that affect spore viability, and of methods used to assess these treatments. Beekeepers became aware of practices that may ultimately be used to decontaminate beekeeping equipment. One high school intern learned laboratory techniques for the examination of spores of the honey bee pathogen we study, and learned about preparations for an oral presentation of the study we worked on.
Publications
- Webster, T.C., Thacker, E.M., Pomper, K., Lowe, J, Hunt, G. 2008. Nosema apis infection in honey bee (Apis mellifera) queens. Journal of Apicultural Research. 47(1):53-57.
- Webster, T.C. and Thacker, E.M. 2008. The effects of selected agents on Nosema apis and Nosema ceranae spore viability. Proceedings of the American Bee Research Conference, Sacramento CA. in American Bee Journal 148(6):562.
- Webster, T. C. 2008. Honey bee. In Encyclopedia of Entomology., second Ed., J Campinera (ed.), Springer.
- Webster, T. C. 2008. Tracheal mite. In Encyclopedia of Entomology., second Ed., J Campinera (ed.), Springer.
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Progress 10/01/07 to 12/31/07
Outputs
OUTPUTS: Methods for the treatment and analysis of Nosema apis and Nosema ceranae spores for viability, and the initial results, were shared with other researchers with related interests: at the USDA-ARS Honey Bee Research Laboratory in Beltsville, MD; at the Agri-Canada Honey Bee Research Laboratory in Beaverlodge, Alberta; and at the Regional Apicultural Center in Guadalajara, Spain. Spore samples from Nosema biotypes were shared with researchers at the Beltsville, MD and Beaverlodge, Alberta laboratories.
PARTICIPANTS: Thomas C. Webster is the p.I. on this project, conceiving and directing it. Etta M. Thacker was the Research Assistant. She prepared and treated samples, recorded data, conducted literature searches, and related activities. Dr. Jeff Pettis, researcher at the USDA-ARS Honey Bee Research Laboratory in Beltsville, MD shared spores of Nosema species with us. Dr. Steve Pernal, researcher at the Agri-Canada Honey Bee Research Station in Beaverlodge, Alberta also shared spores of Nosema species with us. Dr. Mariano Higes at the Regional Apicultural Center in Guadalajara, Spain provided literature, methods and other details pertaining to the study of Nosema species.
Impacts
Observations of Nosema apis and Nosema ceranae spore viability after treatment with selected chemical and physical agents show which might be most appropriate for practical beekeeping when contaminated hive equipment must be disinfected. Most promising at this point is the use of short-wavelength ultraviolet light. Short exposure periods of up to 10 minutes killed up to 100% of spores under laboratory conditions. Similarly, 60 degrees C (140 F) for 2 hours killed many spores of both Nosema species. A staining technique that indicates spores with ruptured membranes was tested and used successfully to measure the effects of the treatments. Both heat and ultraviolet light are likely candidates for use by beekeepers because they are relatively safe, inexpensive and do not leave chemical residues in beeswax or honey, as many antibiotics do. Other treatments which included alcohols, oxidizing agents and fumigants showed some promise and may be most effective when applied
over long time periods. Further study will improve understanding of efficacy and appropriate use by beekeepers.
Publications
- No publications reported this period
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