Progress 06/01/07 to 05/31/11
Outputs
OUTPUTS: Beekeepers continue to have serious winter losses of around 30% of their hives each year on a national basis. Current research indicates that pesticides may be interacting with pathogens and other factors affecting bee health. Our research focuses on identifying and quantifying possible routes of bee exposure to pesticides. In this project, we investigated the potential exposure of honey bees to pesticides in the pollen they collected over the season in several sites in Connecticut, and also in the pollen and nectar of squash plants treated with systemic insecticides. We have now collected pollen for five years, from 2007 to 2011, and analyzed these pollen samples through 2010. Samples collected and analyzed: 2007, 101 pollen samples were taken from hives in 4 locations. Each sample was analyzed individually. In 2008, 124 pollen samples from hives in 3 locations, and grouped into 44 composite samples. In 2009, 123 samples from hives in 5 locations and grouped into 48 composites. In 2010, 164 samples from 4 sites, grouped into 62 composites. In 2011, we reduced the extent of sampling to 46 samples from 2 sites. The samples were extracted using a modified QuECHERs procedure using an isotopically-labeled internal standard. Extractions were made using acetonitrile with solid phase dispersants. The extracts were analyzed using HLPC/MS/MS. In addition, we cooperated with our State Apiary inspector in investigating causes of sudden bee die-offs in Connecticut. Disssemination: Over the course of this project, the results have been formally presented in 26 talks, 3 displays or posters, 5 interviews, and 7 meetings with fellow scientists and stakeholders. PARTICIPANTS: Individuals: Kimberly A. Stoner, Brian D. Eitzer, Ira Kettle, Tracy Zarrillo, Morgan Lowry at CAES. Partner organizations: Natural Resources Conservation Service of CT, Connecticut Beekeeping Association, Backyard Beekeepers Association, Eastern CT Beekeepers Association, US Environmental Protection Agency TARGET AUDIENCES: Beekeepers and beekeeping organizations, researchers on bees, environmental chemists, growers of crops pollinated by bees, conservation organizations, landscapers and groundskeepers, gardeners, government officials, general public concerned about pollinators and pollination. PROJECT MODIFICATIONS: Nothing significant to report during this reporting period.
Impacts
In working with our State Apiary Inspector to investigate honey bee die-offs, we have not found evidence of direct involvement of pesticides in die-offs in our state. In some cases, our colleague Douglas Dingman has found evidence of Nosema ceranae in dying colonies, but was not able to determine if that was the primary cause of colony death. In our analysis of pollen samples, all but two samples had detectable levels of pesticide residues. The mean number of pesticides detected per sample in each year was: 2007 - 4.3, 2008 - 5.4, 2009 - 5.8, 2010 - 4.0. The most frequently detected pesticides were: coumaphos (insecticide, also used as an acaracide by some beekeepers), carbaryl (insecticide), phosmet (insecticide), atrazine (herbicide), imidacloprid (insecticide), dithiopyr (herbicide), pendamethalin (herbicide), and carbendazim (fungicide). Other pesticides, such as the fungicide myclobutanil, were found infrequently, but at high concentrations (up to 4190 ppb) when present.
Publications
- Aronstein, K., F. Drummond, B. Eitzer, J. Ellis, J. Evans, N. Ostiguy, S. Sheppard, M. Spivak & K. Visscher 2011. THE CAP STATIONARY APIARY PROJECT: COLONY STRENGTH DATA ANALYSIS 2009-2010. American Bee Journal 151(5):507-508.
- Krupke, C., B. Eitzer & G.J. Hunt 2011. POTENTIAL ROUTES OF EXPOSURE TO HONEY BEES FROM NEONICOTINOID CORN SEED TREATMENTS. American Bee Journal 151(5):512
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Progress 01/01/10 to 12/31/10
Outputs
OUTPUTS: In 2010, the state apiary inspector investigated sudden die-offs of 31 honey bee colonies owned by 9 beekeepers in 9 different towns. In 2010, pollen samples were collected in 4 locations from early April to early October, completing 4 years of pollen monitoring. Samples from 2007-2009 have been analyzed, and 2010 sample analysis is in progress. In the 2nd year of our study of the movement of neonicotinoid insecticides into pollen and nectar of squash, two insecticides, imidacloprid (Admire Pro) and thiomethoxam (Platinum) were applied to squash grown on black plastic at labeled rates by two different methods: spraying and incorporating in the seed holes before planting, and through drip irrigation 16 days after planting. All samples are analyzed using a multi-residue extraction technique known as QuEChERS (for: quick, easy, cheap, effective, rugged, and safe). In brief, samples are extracted with acetonitrile and cleaned up with solid-phase adsorbents. The extracts are analyzed by high performance liquid chromatography/mass spectrometry/mass spectrometry (HPLC/MS/MS). The described procedures allow us to analyze over 100 different pesticides in a single pollen sample at the parts per billion concentration level. Dissemination: Drs. Stoner and Eitzer presented a paper, "Pesticide in Pollen Trapped from Honey Bee Hives in Connecticut - Two Years of Results in Relation to Acute Toxicity Data," at the American Bee Research Conference on Jan. 15. Dr. Stoner presented a display on "Bees at the Connecticut Agricultural Experiment Station" as part of the Connecticut Groundskeepers Association at the Mohegan Sun in Uncasville on Jan. 29. Dr. Stoner spoke on " Threats to the Health of Wild Bees and Honey Bees" at the Whitney Center in Hamden at the invitation of their Environment Committee on Feb. 4. On Feb. 25, Dr. Stoner spoke on "Plight of the Bumble Bee - Threats the Health of Wild Bees and Honey Bees" at the Ecological Landscaping Conference in Springfield, MA. Dr. Stoner presented a talk on "Threats to the Health of Wild Bees and Honey Bees" at the CT NOFA Winter Conference at Manchester Community College on March 6.On April 20, Dr. Stoner organized and led a day-long meeting of researchers (including Dr. Eitzer) and stakeholders from across New England on Pollination of Northeastern Specialty Crops at the University of Massachusetts. This meeting was funded by the Northeastern Regional Association of State Experiment Station Directors. Dr. Stoner made a presentation "Why Pollinator Decline is Important" to the Wild Ones (a native plant group) at Connecticut College in New London on May 1. Dr. Stoner spoke to the Naugatuck Audubon Society on "Threats to the Health of Wild Bees and Honey Bees" at the Kellogg Environmental Center in Derby on May 18.Dr. Stoner spoke about "Pollinators in the Landscape" to the Annual Meeting of the NOFA Organic Land Care Program at the University of Connecticut on Dec. 7.Dr. Stoner was interviewed about honey bee colony losses by reporters from the Connecticut News Service, the Greenwich Time, and the Waterbury Republican American on July 19, July 19, and July 26, respectively. PARTICIPANTS: Individuals: Kimberly A. Stoner, Brian D. Eitzer, Ira Kettle, Tracy Zarrillo, Morgan Lowry at CAES. Partner organizations: Natural Resources Conservation Service of Connecticut, Connecticut Beekeeping Association, Backyard Beekeepers Association, Eastern Connecticut Beekeeping Association TARGET AUDIENCES: Beekeepers and beekeeping organizations, researchers on bees, environmental chemists, growers of crops pollinated by bees, general public concerned about bees, landscapers and groundskeepers, gardeners, conservation organizations, govermnent officials PROJECT MODIFICATIONS: Nothing significant to report during this reporting period.
Impacts
Over the four years that we have been monitoring pesticides, we have found that the pesticide levels in pollen are dependant on both time and location. All of the pollen samples tested have had pesticide residues, with the average number of residues per sample ranging from a low of 4.3 in 2007 to a high of 6.9 in 2009. This later number, however, is skewed to the high side because the orchard hive location in 2009 had more pesticide residues than were observed in the other samples (12.4 residues per sample at the orchard as compared to 5.9 residues per sample at the other four locations). Over the four years that we have been monitoring pesticides, we have found that the pesticide levels in pollen are dependant on both time and location. In our analysis of movement of neonicotinoid insecticides into the pollen and nectar of squash, we found that both imidacloprid and thiamethoxam residues were present in the nectar and pollen of treated plants, regardless of whether the systemic insecticides were applied to the soil before seeding or applied to transplants through drip irrigation.
Publications
- Stoner, K.A. and B. D. Eitzer. 2010. Pesticide in Pollen Trapped from Honey Bee Hives in Connecticut-Two Years of Results in Relation to Acute Toxicity Data. American Bee Journal 150 (5): 510.
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Progress 01/01/09 to 12/31/09
Outputs
OUTPUTS: Honey bees (Apis mellifera) have died off in large numbers in recent years. A whole range of stresses are being investigated, including exposure to pesticides, the stressor we are studying here. Activities: In 2009, a total of 123 pollen samples were taken from honey bee hives in 5 locations and grouped into 48 composite samples. These samples have all been analyzed for pesticides. In 2008, a total of 124 pollen samples were taken from 3 hives and grouped into 44 composite samples. The analyses of these samples were completed in 2009. In our experimental study of movement of systemic neonicotinoid insecticides in summer squash, two insecticides, imidacloprid (Admire Pro) and thiomethoxam (Platinum) were applied to summer squash grown on black plastic at labeled rates by two different methods: spraying and incorporating in the seed holes before planting, and through drip irrigation 16 days after planting. All samples are analyzed using a multi-residue extraction technique known as QuEChERS (for: quick, easy, cheap, effective, rugged, and safe). In brief, samples are extracted with acetonitrile and cleaned up with solid-phase adsorbents. The extracts are analyzed by high performance liquid chromatography/mass spectrometry/mass spectrometry (HPLC/MS/MS). The described procedures allow us to analyze over 100 different pesticides in a single pollen sample at the parts per billion concentration level. Dissemination: Dr. Stoner presented a paper, "Measuring Pesticides in Pollen" at the Atlantic Coast Agricultural Conference in Atlantic City, NJ on Jan. 14. Dr. Eitzer presented a paper, "Measuring Pesticide in Pollen Trapped from Honey Bee Hives in Connecticut" at the American Bee Research Conference in Gainesville, FL on Feb. 5. Drs. Stoner and Eitzer displayed a poster on measuring pesticides in pollen at Agriculture Day at the Capitol in Hartford, CT on Mar. 18. Drs. Stoner and Eitzer participated in a meeting on the research needed to understand the potential effects of pesticides on bees at the Natural Resources Defense Council in Washington, DC on Mar. 24. Drs. Stoner and Eitzer made a presentation, "Measuring Pesticides in Pollen" to the Connecticut Beekeeping Association in New Haven, CT on Apr. 4. Dr. Eitzer presented a seminar on "Pesticide Residues in Honey Bee Pollen" at the University of Massachusetts, Amherst on April 7, 2009. Dr. Stoner presented a talk on "Threats to the Health of Honey Bees and Wild Bees" to the Hamden Land Trust in Hamden, CT on June 17. Dr. Stoner participated in a meeting held by USDA-APHIS and US EPA on measuring the effects of pesticides on honey bees in Alexandria, VA on July 8-9. Dr. Stoner presented a talk on "Threats to the Health of Honey Bees and Wild Bees" to the Cheshire Garden Club in Cheshire, CT on Sept. 14. Dr. Stoner presented a poster on the Station's work on honey bees, including the work on pesticides and pollen, at the Southern New England Bee Assembly in Hamden, CT on Nov. 21. Dr. Eitzer presented a paper on "Determination of Pesticide Residues in Honey Bee Pollen by HPLC/MS/MS" at the Society of Environmental Toxicology and Chemistry 30th annual meeting in New Orleans, LA on Nov. 20. PARTICIPANTS: Individuals: Kimberly A. Stoner, Brian D. Eitzer, Ira Kettle, Tracy Zarrillo, Morgan Lowry at CAES. Partner organizations: Connecticut Beekeeping Association, Backyard Beekeepers Association, Eastern Connecticut Beekeeping Association TARGET AUDIENCES: Beekeepers and beekeeping organizations, researchers on bees, environmental chemists, growers of crops pollinated by bees, general public concerned about bees, gardeners, conservation organizations, govermnent officials PROJECT MODIFICATIONS: Nothing significant to report during this reporting period.
Impacts
Change in Knowledge: Over the three years that we have been monitoring pesticides, we have found that the pesticide levels in pollen are dependant on both time and location. All of the pollen samples tested have had pesticide residues, with the average number of residues per sample ranging from a low of 4.3 in 2007 to a high of 6.9 in 2009. This later number however is skewed to the high side because the orchard hive location in 2009 had more pesticide residues than were observed in the other samples (12.4 residues per sample at the orchard as compared to 5.9 residues per sample at the other four locations). In analysis of whole plant samples of summer squash treated with neonicotinoid insecticides, we found that whole plants treated through drip irrigation had higher concentrations of insecticides in the tissues than whole plants treated by spraying the seed holes. Thiomethoxam averaged 362 ppb in the drip irrigation samples as compared to 154 ppb by spraying and incorporating in the seed holes, while imidacloprid averaged 218 ppb in the drip irrigation samples and 47 ppb in the spraying and incorporating into seed hole samples.
Publications
- Stoner, K.A. and B.D. Eitzer 2009. Measuring Pesticides in Pollen. In Proceedings of the Atlantic Coast Agricultural Conference. p. 86.
- Stoner, K.A. and B.D. Eitzer 2009. Measuring Pesticides in Pollen Trapped from Honey Bee Hives in Connecticut in Proceedings of the American Bee Research Conference 2009, published in American Bee Journal, 2009, 149(6), 588-589.
- Eitzer, B.D. and K.A. Stoner. 2009. Determination of Pesticide Residues in Honey Bee Pollen by HPLC/MS/MS in the Abstract Book of the Society of Environmental Toxicology and Chemistry 30th Annual Meeting p.36
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Progress 01/01/08 to 12/31/08
Outputs
OUTPUTS: Activities: We collected pollen pellets from the honey bees using a pollen trap, a series of screens that the returning forager bees must pass in order to get back into the hive. The trap knocks the pollen pellets off the hind legs of the bee and collects the pellets in a drawer. We used a pollen trap mounted at the bottom of the hive. We collected the pollen twice a week, from four locations in CT in 2007 and 3 locations in 2008: 1. at our offices in New Haven on the edge of the city, 2. at our experimental farm surrounded mostly by suburbs, 3. at an orchard on the edge of a suburb during the blooming season of apples and blueberries in 2007 only, and 4. at another suburban site on the edge of a large agricultural area growing vegetable crops. Pollen samples were analyzed by a multi-residue technique known as QuEChERS, (for: quick, easy, cheap, effective, rugged and safe). The samples were treated with a series of solvents to separate the pesticides from the rest of the pollen, and the extracts were analyzed by high performance liquid chromatography/mass spectrometry (HPLC/MS). To enhance the specificity of analysis, the ion yielded by HPLC/MS is fragmented again in a technique known as MS/MS. This allows us to measure imidacloprid to a detection limit of 0.1 parts per billion (PPB), other neonicotinoids to a detection limit of 0.5 - 2 PPB, and most other pesticides we will be reporting here to 1 - 10 PPB. Dissemination: Dr. Stoner presented a talk, "Threats to the Health of Bees," at the Environment and Conservation forum at Heritage Village, Southbury, CT on Jan. 16. Dr. Stoner presented "Threats to Honey Bee Health" at the Cultivating an Organic Connecticut Conference of the Northeast Organic Farming Association in Windsor, CT on March 8. Drs. Stoner and Eitzer presented a poster on "Honey Bee Exposure to Pesticides" at the Connecticut Conference on Natural Resources, University of Connecticut, Storrs on March 10. Drs. Stoner and Eitzer spoke at the Spring Meeting of the Connecticut Beekeepers Association on "Measuring Pesticides in Pollen, " New Haven, CT, April 5. Drs. Eitzer and Stoner presented a barn exhibit, "Analysis of Pesticides in Pollen Collected by Honey Bees" at Plant Science Day, Hamden, CT on Aug. 6. Dr. Stoner presented a workshop on "Threats to the Health of Bees" at the NOFA Summer Conference at the University of Massachusetts in Amherst on Aug. 9. Dr. Eitzer gave a presentation entitled "Pesticide Residues in Pollen Collected by Foraging Honeybees" at the national meeting of the American Chemical Society in Philadelphia, PA on Aug 17. Dr. Stoner presented research on pesticides in pollen the Back Yard Beekeepers Association in Westport, CT on Sept. 30. Dr. Stoner presented a talk, "Pesticides in Pollen Collected from Honey Bee Hives in Connecticut," at the Pollinator Conference: Information for Action, sponsored by the U.S. Fish and Wildlife Service and held at the University of Massachusetts in Amherst on Oct. 3. Dr. Eitzer presented a talk, "Analysis of Pesticide Residues in Honey Bee Pollen by Liquid Chromatography/Mass Spectrometry" at the Eastern Analytical Symposium & Exposition in Somerset, NJ on Nov. 18, 2008 PARTICIPANTS: Individuals working on the project: in addition to Dr.Kimberly Stoner, Dr. Brian Eitzer, and State Apiary Inspector Ira Kettle, we have also collaborated with Dr. Phillip Lewis of USDA-APHIS. Partner organizations include the Connecticut Beekeepers Association, the Back Yard Beekeepers Association, and Project Apis m. TARGET AUDIENCES: There are many audiences interested in this research project. In 2008, we made presentations to several beekeeping groups, farming organizations, naturalists concerned about pollinator species, colleagues in entomology and analytical chemistry, and the general public. PROJECT MODIFICATIONS: Not relevant to this project.
Impacts
Change in Knowledge: All samples of the 2007 collection have been analyzed, the 2008 samples are currently undergoing analyses. In the 2007 pollen samples we detected 37 pesticides: 15 insecticide/acaricides, 11 fungicides, 10 herbicides, and 1 plant growth regulator. All samples had at least one pesticide detected, and the mean number of detections was 4.25 pesticides per sample. The most commonly detected pesticide was coumaphos, which had definitely been used in the hives in sites 1 and 2 as a miticide in the previous year. (Hives in sites 3 and 4 belonged to another beekeeper.) Carbaryl and phosmet, both of which are considered highly toxic to bees, were the most commonly detected field pesticides. Imidacloprid was detected 30 times, mostly at low levels. The pesticides found at the highest levels were both fungicides: myclobutanil, which belongs to the class of fungicides found to act synergistically to increase the effects of pyrethroids and certain neonicotinoids on bees, and boscalid. Carbaryl was the only pesticide found both frequently and at relatively high levels at all sites. Imidacloprid was found at a high level (70 ppb) in a single sample - all other samples were at 3.4 ppb or below. We are in the process of separating this sample into components based on the color and texture of the pollen pellets, analyzing the components, and identifying the pollen in the different pellet types in order to determine the plant sources. We also found that thiamethoxam was present in the flowers of plants that had been grown with treated seed indicating that bees could get exposed to this neonicotinoid pesticide while foraging.
Publications
- Stoner, K.A. and B. Eitzer. 2008. Pesticides in Connecticut Pollen. Abstract and Powerpoint presentation. Pollinator Conference: Information for Action, Oct. 3-4, 2008, University of Massachusetts, Amherst. Online at: http://www.millersriver.net/pollen/
- Eitzer, B.D. and K.A. Stoner. 2008. Analysis of Pesticide Residues in Honey Bee Pollen by Liquid Chromatography/Mass Spectrometry. Abstract Book of the 47th Eastern Analytical Symposium & Exposition, November 17-20, 2008, Somerset, NJ p.55.
- Eitzer, B.D. and K.A. Stoner. 2008. Pesticide Residues in Pollen Collected by Foraging Honey Bees. Abstract in Picogram v. 75 Proceedings of the Agrochemical Division of the American Chemical Society 236th National Meeting, August 17-21, 2008, Philadelphia, PA p.75.
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Progress 06/01/07 to 12/31/07
Outputs
OUTPUTS: During the past year, we have been working on the development of a multi-residue method for the analysis of pesticides in bee pollen. Our goal has been a method that can be used to quantify the neonicotinoid pesticides at concentration levels in the low parts per billion range. We have focused on the neonicotinoid pesticides as these are of greatest concern to the beekeeping community. At the same time, we are trying to keep the method as broad as possible so that other pesticides can be monitored. We have settled on a modified version of the QuEChERS (for Quick, Easy, Cheap, Effective Rugged, Safe) method. In brief, a sample of bee pollen is rehydrated with water, spiked with internal standards, extracted with acetonitrile, cleaned up with solid phase dispersal reagents, solvent exchanged to toluene and concentrated. The final extract is analyzed by liquid chromatography mass spectrometry using positive ion electrospray. With this technique we have demonstrated that less
than 1 PPB of imidacloprid can be detected.
PARTICIPANTS: Dr. Kimberly Stoner is the principal investigator collecting samples from honey bee hives and working with beekeepers and beekeeping organizations. Ira Kettle is the state apiary inspector, working with beekeepers and assisting with outreach for this project. Dr. Brian D. Eitzer is the principal investigator working on the development of analytical methods and analysis of samples for this project.
TARGET AUDIENCES: Target audiences are beekeepers and beekeeping organizations, growers of crops that require insect pollination, and the general public. We have conducted outreach to beekeepers through meetings of the Connecticut Beekeepers Association, Backyard Beekeepers Association, and the Southern New England Bee Assembly.
Impacts
This analytical procedure has been used to analyze pollen samples that have been periodically collected from 4 hives within the State of Connecticut. These hives represent urban, suburban and rural locations allowing us to examine pollen pesticide loads for both seasonal and site differences. To date a total of 86 samples have been collected with 48 of the samples having been analyzed. Over 25 different active pesticide ingredient or metabolites have been detected in these samples with concentrations that can range from less than 1 PPB to grater than 1 PPM. Imidacloprid, the pesticide of greatest interest has been found in approximately half of the samples analyzed. We have found that the concentrations of pesticides in pollen are highly variable both by location and time.
Publications
- No publications reported this period
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