PRODUCTION OF NATIVE BUMBLE BEES FOR POLLINATION OF WEST COAST CROPS

• Sponsoring Institution: National Institute of Food and Agriculture
• Project Status: COMPLETE
• Funding Source: SMALL BUSINESS GRANT
• Reporting Frequency: Annual
• Accession No.: 0225297
• Grant No.: 2011-33610-30442
• Proposal No.: 2011-00022
• Project Start Date: Jul 15, 2011
• Project End Date: Oct 31, 2012
• Grant Year: 2011
• Program Code: [8.2]- Plant Production and Protection-Biology

Recipient Organization
Mite Bee Farm, Inc.
30807 Decker Ridge Road
Corvallis,OR 97333

Performing Department
(N/A)

Non Technical Summary
Farmers in the USA have largely depended on the European honey bee, Apis mellifera, for crop pollination. In recent years, Varroa mites, Tracheal mites, Nosema, Viruses and Colony Collapse Disorder have reduced the availability of honey bee colonies. This has created a critical need for additional managed pollinators. Mason bees, leaf cutter bees and bumble bees are being managed to varying extents. Of these, bumble bees have the best economic potential as pollinators. They are exceptional pollinators for crops such as blueberries that require buzz pollination and for crops raised in greenhouses where honey bees are not effective. The US has over 40 bumble bee species but only one species, B. impatiens, is available commercially. This species is endemic to the Midwest and the East, and is thus not available to growers in western states (such as Oregon) that do not permit the introduction of non-native bees due to concerns about pathogens. Nationwide concerns about bumble bee declines have led scientists to petition the USDA to ban movement of B.impatiens to the entire west coast. An urgent need for commercial production of west coast bumble bee species exists. In western Oregon, several native bumble bee species are thriving but timing, numbers & consistency are not manageable with wild bumble bees. Bumble bee species endemic to western US have been successfully raised in captivity for research purposes but tactics need to be developed for effective and economical commercial production. The objectives of the current research proposed are: 1) Development of techniques for breaking diapause and enhancing nest initiation by queens, and growth and development of colonies; 2). Comparison of pathogen load in wild and captive-reared bumble bees, and 3). Evaluation of tactics for overwintering queens. The research will focus on three west coast species, B. vosnesenskii, B. mixtus, and B. nevadensis, and will be conducted by Mite Bee Farm, Inc. and Oregon State University (OSU). Mite Bee Farm, Inc., a woman-owned small business, is currently engaged in honey bee queen production will diversify its operations by the addition of bumble bee rearing. A research associate with experience in bumble bee rearing from OSU will be hired for support in achieving the objectives above. The techniques and products developed by Mite Bee Farm, Inc. from this research will greatly enhance commercialization of west coast bumble bee species for producers of a great diversity of fruit and vegetable crops. The project is aligned with USDA AFRI priority of Global Food Security and Hunger as new technologies will be developed that will enhance production of foods for humans and livestock. It will provide valuable data for addressing the petition submitted to USDA related to restriction of movement of B. impatiens to the west for reducing risks associated with pathogen movement. If alternative west coast bumble bee species are available commercially, the ban, if implemented, will not have negative economic impacts on west coast producers of bee-pollinated crops.

Animal Health Component

30%

Research Effort Categories

Basic

40%

Applied

30%

Developmental

30%

Classification

Knowledge Area (KA)	Subject of Investigation (SOI)	Field of Science (FOS)	Percent
211	3090	1130	100%

Knowledge Area
211 - Insects, Mites, and Other Arthropods Affecting Plants;

Subject Of Investigation
3090 - Other Pollinators;

Field Of Science
1130 - Entomology and acarology;

Keywords

bumble bee

rearing

pollination

overwintering

Goals / Objectives
The objective of this proposal is to develop tactics for commercial production of healthy colonies of west coast bumble bees. Specific objectives include: 1.Development of techniques for breaking diapause and enhancing nest initiation by queens, and growth and development of colonies.2. Comparison of pathogen loads in wild and captive-reared bumble bees. 3. Evaluation of tactics for overwintering queen bumble bees. Experiments under Objective 1 will be initiated soon after collection of bumble bee queens from early spring through summer. Objective 2 will be ongoing throughout the duration of the project as individuals will be collected from the captive reared colonies and in the wild. Objective 3 will be initiated in late summer and continue until the duration of the project. The project aims to deliver an alternative pollinator to several west coast fruit and seed crops such as blueberries, cranberries, red clover seed and greenhouse vegetables including tomatoes and peppers. The current project will provide growers with access to commercial bumble bee populations that can be placed in their fields during bloom. Currently, a petition has been submitted to the federal agency, USDA, requesting a ban on movement of B. impatiens to the west where it is not endemic. The project will provide valuable data to USDA, in their efforts to assess the situation and determine if a ban is to be established for restriction of movement of B. impatiens to west coast growers. Concerns about economic impacts on west coast growers if the ban is established will be mitigated when west coast bumble bees are available commercially.

Project Methods
B. vosnesenskii, B. mixtus and B. nevadensis are the native, western bumble bee species under study. Typically, queens are exposed to CO2 for up to 30 minutes to stimulate egg laying but the impact of CO2 on western native species is unknown. CO2 experiments in closed containers will be conducted thus: A. No CO2, B. 3 minutes CO2, C. 20 minutes CO2. Queens will be evaluated daily for 30 days for survivorship and egg laying. The experiment is a randomized block design with 10 replicates. Data will be analyzed using ANOVA and Tukey's multiple comparison test. Nest initiation in captured bumble bees can take several days. There is also a negative correlation between the number of days to emergence of the first worker and ultimate colony size. Egg laying can be initiated sooner by the presence of other bees. Bees and materials will be used to mimic bumble bee pupae as queens deposit egg clumps on or near pupae. Several techniques will be compared: A. One queen bumble bee, B. Two queen bumble bees, C. One queen bumble bee and one honey bee worker, D. One queen bumble bee and one bumble bee worker, E. Foam cocoon, F. Foam cocoon wrapped in silkworm silk. The experiment is a randomized block design with 10 replicates. Egg clumps, queen mortality, successful colony establishment, and time to initiate egg laying will be recorded . Data will be analyzed using ANOVA and Tukey's multiple comparison test to determine the best strategy. Confined colonies produce only 1 egg clump per day but free flying colonies produce 2 per day. Colony development will be compared in the rearing room and in the greenhouse. Following the production of the first workers, some colonies will be transferred to the greenhouse to forage freely and others will remain in the rearing room. Colony size and weight of workers will be observed twice weekly. The standard t-test will be used for analysis of this data. Pathogen loads can be higher in colonies raised in captivity. Nosema bombi (fungus/microsporidian), Crithidia bombi (protozoa), Sphaerularia bombi (nematode) are the study pathogens. 10 wild bumble bee colonies of each species will be collected and randomly selected workers preserved at -40C. 10 captive-reared colonies will also be sampled as such. Samples will be dissected for analysis by microscopy. Pathogens will be identified and scored for levels of infection. Queens have been observed to use soil, peat and sawdust for overwintering. These substrates will be evaluated with modifications to minimize queen disturbance. Observations will be made on the numbers of queens that use each substrate. Further, overwintering queens will be observed weekly for 3 months to evaluate the impact of temperature on vitality and mold development. Queens from the previous experiment in their selected substrate will be transferred to the following temperatures: A. 2C, B. 4C, C. 8C. The experiment is a randomized block design with 10 replicates. Data will be analyzed using ANOVA and Tukey's multiple comparison test for identification of the most efficient and economical strategy. This data will serve bumble bee producers for greenhouse pollination and crops not suited to honey bees.

Progress 07/15/11 to 10/31/12

Outputs
OUTPUTS: We focused on the technical objectives as listed in the research proposal: 1. Development of techniques for breaking diapause and enhancing nest initiation by queens, and growth and development of colonies, 2. Comparison of pathogen load in wild and captive-reared bumble bees, 3. Evaluation of tactics for overwintering queens. We completed many experiments focusing on breaking diapause of newly emerged queens in the spring. Three species of Bombus (vosnesenskii, nevadensis, and griseocollis) that were collected in high enough numbers (70 queens per species) to follow through with experiments were the species investigated. The first experiment was to try different rearing set ups to see what would encourage queens to start laying eggs as soon as possible after being placed in the rearing facility. Queens were set up alone, alone with a foam blob, alone with a foam blob that had been dipped in honey bee wax, with another queen of the same species, with a worker of the same species or with young honey bee workers. This was followed with documenting how successful queens were with rearing eggs through to new workers and how well the colony continued to grow. We compared pathogen loads in queens that were under rearing conditions for several weeks to those recently collected from the wild. One hundred fifty queens of four species: Bombus vosnesenskii, B. griseocollis, B. nevedensis and B. mixtus were used for this investigation. An emphasis was placed on the pathogens that commonly infect bumble bees: Nosema bombi, Crithidia bombi, Apicystis bombi and Sphaerularia bombi. We dissected bumble bees and did visual screening for these pathogens. Notes were made of yeast, bacteria and other fungal pathogens when observed. Colonies in the rearing room were compared with colonies in the greenhouse. Queens were collected after mating were placed in the refrigerator at 4 degrees Celsius in small containers in October 2011. They were examined for survival in February and March 2012. Results of this work were reported at Pacific Branch Entomology Society of America meeting Portland, OR. March 28, 2012 in the symposium The Changing Face of Pollination in Western Agriculture. PARTICIPANTS: Oregon State University, Crop and Soil Science Department: Dr. Rao's laboratory did the microbiological work on the queen honey bees. Post Doctoral Fellow: Helped design the experiments and trained a worker and myself in the art of rearing bumble bees. Worker: who did the care of the bumble bees . Worker who oversaw building of materials and equipment, including the construction of the greenhouse. TARGET AUDIENCES: Report was made at the Pacific Branch Entomology Society of America's meeting 2012. Providing information regarding bumble bees to other bee researchers. PROJECT MODIFICATIONS: Not relevant to this project.

Impacts
We worked with five species of Bombus native to Oregon, specifically the Willamette Valley. Of the five three were outstanding and will provide focus for further studies. The results with queens over the six treatment sets were not consistent in 2011 and during work in 2012 queens were placed alone. These queens showed a good response similar to successes for trails in 2011. The advantage of queens alone in their rearing containers is both saving of time and resources especially a second queen. Queens kept in the rearing room through emergence of their first and second brood of workers did well with the transfer to the greenhouse. Colonies left in the rearing room after worker emergence tended to have problems with wax moths while those moved into the greenhouse developed normally with few or no wax moths. All the bees moved into the greenhouse were active within the space, and from dawn until dusk the activity of bees working flowers or visiting feeding stations was audible. There was a mix of flowers available to the bees: tomato, anise hyssop, sage, basil, catnip, borage to name a few as well as feeding stations with sugar syrup and feeding stations with pollen. All these plants produced seeds or fruit. Fifty percent of the B. vosnesenskii and griseocolis colonies in the greenhouse produced males and queens. B. vosnesenskii colonies were overall larger in population of workers than either B. nevadensis or B. griseocolis. The later two species would be good candidates for greenhouse pollination because their smaller colonies would work better in smaller spaces without as much conflict with human employees that come into greenhouses to water or pick produce. Of the mated queens collected and placed into small containers that went into the refrigerator only B. vosnesenskii survived. Of the three Bombus species mentioned above the only species to have the nematode Sphaerularia bombi was B. vosnesenskii. The only microorganism seen in consistently higher numbers in captive queens was yeast.

Publications

• No publications reported this period

Progress 07/15/11 to 07/14/12

Outputs
OUTPUTS: The experiments regarding the days required for spring caught queens to begin oviposition were completed. The experiments following the queens from first egg laying to production of workers were completed. Treatments for days required to first ovipostion and the nest initiation experiments were single queens, two queens of the same species, a queen with a worker of the same species, a queen bumble bee with several day old honey bee workers, a queen with a foam knob(simulating an early nest pollen ball with egg clump, a queen with foam knob covered with honey bee wax. The experiments to stimulate nest initiation using carbon dioxide treatments were completed. The Bombus species used were B. griseocollis, B. vosnesenskii, B. nevadensis. Disease prevalence for all three species continues to be monitored. In the fall of 2011 some queens were mated and then successfully overwintered. PARTICIPANTS: Lynn A. Royce PI spent 51% FTE overseeing and managing the project as well as helping with the work. Bee specialist spent 49% FTE checking on colonies and doing other work including upkeep of equipment and overseeing greenhouse construction. Post Doc spent 100% of her time in catching bumble bee queens, overseeing the details of the experiments and collecting and entering the data. A helper was paid hourly to collect bumble bee queens and males, do necessary dish washing and cleaning, bumble bee feeding. TARGET AUDIENCES: Nothing significant to report during this reporting period. PROJECT MODIFICATIONS: Not relevant to this project.

Impacts
The three species used in the above experiments show promise as pollinators for Oregon crops. Oregon has state laws that prohibit importation of bees other than honey bees from locations outside the state. Therefore, these bumble bee reared locally in Oregon can provide pollination for early and greenhouse crops that cannot be pollinated by honey bees. While all three could be used for greenhouse crops B. nevedensis and B. grisicollis produce smaller colonies and may therefore be easier to manage in a confined space. When funds were made available 2011 we were able to construct a bee tight greenhouse where we could fly virgin queens and males for mating experiments and ultimately overwintering. Conversations with farmers indicate a varying bloom time for crops needing pollination especially for early crops and greenhouse crops. These conversations will help in the design of future overwintering work and experiments so that we can have bumble bee colonies of pollination size for early blooming crops. Working with nest initiation experiments suggest queens are sensitive to vibration and disturbance that may lead to failure to initiate a nest. Therefore room renovation were undertaken to lower vibration and disturbance. This improved our ability to get nest initiation using a single queen making rearing more efficient.

Publications

• No publications reported this period