2013 OBA Tech-Transfer Program Research Projects/Programs

2013 Research and Activities
Les Eccles, Melanie Kempers, Devan Rawn & Brian Lacey
Funding sources are provided in brackets
TTP = OBA Tech-Transfer Program
CAAP = Canadian Agricultural Adaptation Program
OMAFRA = special funding from OMAFRA
KTT = OMAFRA/U of G Knowledge, Translation and Transfer Program

A. Breeding and Maintaining Parasitic Mite Resistant Honey Bee Stocks in Ontario

1. Maintenance of the Hygienic Trait in Ontario Bee Stocks (TTP)

The Ontario Honey Bee Breeding Program is a long term program to incorporate and maintain mite resistant characteristics in Ontario’s honey bee stock. 2006 was the 15th year of testing for tracheal mite resistance and 2013 was the 16th year of testing for hygienic behaviour. Bee breeders in Ontario selected colonies with favourable characteristics to be tested for tracheal mite resistance and hygienic behaviour. The tracheal mite resistance test has not been performed since 2006 due lack of tracheal mite infested colonies to perform the tests on Ontario breeder colonies. Hygienic behaviour is important for mite and disease resistance within a colony. The liquid nitrogen freeze kill method was used to test for hygienic behaviour. Colonies which ranked in Group 1 (>95% of killed brood removed) and Group 2 (75-95% of killed brood removed) were recommended for use as breeders for the subsequent generation. Testing for hygienic behaviour was conducted on 195 potential breeder colonies for 8 bee breeders in 2012.

Comments on 2013 season: - Later testing due to high demand for queens and less time to commit to hygienic testing - Queens sent to PEI from OBBA breeders. - 2 bee breeders with a long history in the breeding program did not participate in hygienic testing due to colony loss and high queen demand.

2. Health Status of Colonies Tested in the Breeding Program (TTP)

Honey bee samples were collected from potential breeder colonies. At the time of hygienic behaviour testing, forager bees were collected. Bees were also collected from the brood chamber before treatments were applied in the fall. Varroa and tracheal mite infestation levels were determined and nosema spore levels analyzed as an indication of the health of the colonies. Monitoring the health of breeder colonies will ensure the quality of the bee stock produced in Ontario. Samples from 195 potential breeder colonies from 8 bee breeders were analysed for nosema, tracheal mite and varroa testing.

3. Survey of the Quality of Honey Bee Queens from Ontario Breeders (CAAP)

Queen Fertility Since the increased overwinter mortality of honey bee colonies in 2007, beekeepers have observed that the queen bee's lifespan, which normally last 2-3 years, has been reduced to only 1 year of production. The concern is that the fertility of honey bee queens is not sufficient to maintain colony which results in weakened and susceptibility to other colony stresses. Monitoring and selection of fertile queens is an important aspect of breeding and producing productive honey bees that will allow beekeepers to have improved consistency in their beekeeping activities. Part of the process to bring fertility testing technology to beekeepers is training technicians to provide the service. This includes setting up the protocol to take samples from the field, and the laboratory procedures that make efficient and cost effective evaluations possible at the industry level. Training, consulting, and use of laboratory facilities for this activity for the OBA Tech-Transfer Program was provided by Dr. Ernesto Guzman’s Apiculture program that the University of Guelph and visiting technician MSc.Megan Taylor from Washington State University. The protocol was adopted and refined from protocol Dr. Guzman and Megan Taylor demonstrated in order to count live and dead semen (Taylor, 2009).

In 2013, eight bee breeders participated and donated 80 queens for fertility testing. The increase in participation was a result of the interest the industry took in the results from the work in 2012. The results show an immediate status of the management of a breeding program and environmental factors that affect the fertility of queens they are producing. The number of potential queens tested in 2013 was likely reduced, because of the high demand for queens by the industry to recover from high overwinter mortality in 2012-2013, which resulted in fewer queens available for testing. There was also interest in having imported queens from Australia tested to ensure the queens Ontario beekeepers are introducing are of sufficient quality. Unintended results showed queens from areas impacted with reports of insecticide kills in 2012 and 2013 had significantly decrease sperm viability compared to queens harvested from areas isolated from the suspect insecticide.These preliminary results have become of interest by the Pesticide Regulatory Management Agency and Ontario Ministry of Agriculture and Food, for the OBA Tech Transfer Program to design and implement research on the effect of specific pesticide on honey bee queen and drone fertility. Training of technicians from the TTP to perform queen fertility testing is important to allow this service to arrive and be adopted by beekeepers in the field. The cost of this service is economical for bee breeders at $30 for the first queen and $20 for every additional queen. Fertility testing will allow beekeepers to select for increased fertility and test for improvements from management of queen mating from this service after the project has been completed; as it will be integrated into a formal breeding program.

4. Development of Bee Breeding Program (CAAP)

Using critiqued information and research, and consultation with experienced researchers and industry individuals; a formal breeding program is being created to answer the demand by queen breeders, to expand the criteria and precision in which they use data to improve their breeding programs. Research, discussion and consulting are important activities to advance and develop this objective. Key contributors to the development of this activity are: Dr. Ernesto Guzman, University of Guelph BSc. Paul Kelly, University of Guelph Dr. Pierre Giovenazzo, University of Laval Members of the Canadian Association of Professional Apiculturists Ontario Bee Breeders Association Ontario Beekeepers’ Association Tech Transfer Program An international event called Apimondia was held in Quebec during Nov. 15-18 2012. The focus of this symposium was “Queen Breeding, Selection and Bee Health”. The OBA Tech Transfer Team was asked to organize and host a focus group session during this symposium, in which 22 highly reputable authorities on queen breeding and selection attended, in order to discuss priorities needed to improve breeding programs used by beekeepers. This event was an extremely unique opportunity for this CAAP project to be presented to an international group of experts in the field of queen breeding and genetic improvement.

The OBA TTP took this opportunity to invite 6 honey bee technicians from Saskatchewan, Quebec, and the United States to take part in facilitating this focus group and begin collaborations between our provinces and countries to improve the focus of bee breeding. This focus group session resulted in improving the criteria and priority setting part of designing a formal breeding program for honey bee characteristics that will satisfy the demand by the beekeeping industry. This information can now be used in the statistic program that is being finalized for beekeepers to use when receiving data from the results for the various characteristics being evaluated. Using all information available from previous research and consulting during this project resulted in a completed program for bee breeders to follow in order to understand the criteria they are selecting for and how to evaluate them. This data can now be entered into a controlled Excel (Microsoft) spreadsheet that allows beekeepers selection compare and make evaluations based on the objectives of their breeding program. This program will also allow breeders to compare their results with other breeders that can be then be used to integrate characteristics from other lines of honey bees into their breeding program. This will be the first program available to the beekeeping industry that will enable bee breeders to pool results and work together in order to improve stock available for the beekeeping industry. This breeding program will be the basis of the Ontario Resistance Honey Bee Selection (ORHBS) program. TTP will work with ORHBS to use this program to build on the success and reputation of Ontario bee breeding and begin a certification program to qualify the participants’ efforts. A new course has been proposed in order to train current and new bee breeders to improve and update their bee breeding program.

B. Integrated Pest Management (IPM) Program to Control Honey Bee Pests & Diseases:
1. Best Management Practices of Pollination Colonies (CAAP)
In 2012 and 2013, 360 colonies were monitored for their pest and disease status in relation to the controlled application of pollen supplement in order to identify the risk to colony health due to malnutrition. It was discovered that diseases nosema ceranea and european foul brood were at a much higher levels in colonies transported to blueberry and cranberry pollination in eastern Canada, compared to colonies that stayed in Ontario for honey production. It is interesting to note that varroa mite, which is the main pest for honey bees were present in much lower numbers in pollination colonies, due to the inability of the mite to reproduce when pollination honey bee colonies are reproducing less due to nutritional stress. This is important information for beekeepers in order to understand the Integrated Pest Management (IPM) beekeepers must implement specifically for pollination services. 2012-2013 overwinter mortality was lower for beekeepers that sent colonies to blueberry and cranberry pollination in 2012, due to lower levels of varroa mites.
A new method to apply oxalic acid to control varroa mite in the spring season for queen/nuc producers and pollination beekeepers was tested. The advantage to this treatment is that there is no effect on honey bee populations or queens and is considered a treatment to prevent resistant building to other commonly used miticides in an IPM program. These results will be submitted to the Ontario Provincial Apiarist to update the treatment recommendations available to beekeepers. In 2013, oxalic acid treatment was applied to nucleus colonies and split colonies to control varroa mites before colonies are sold by queen and nuc producers to their customers. The results of this testing will be integrate into workshops, recommendation and training of beekeepers that want to control varroa without the use of “synthetic chemical” products that can result in resistant varroa mites after repeated use. This will allow the beekeeping industry to depend on the functionality of these chemical varroa mite treatments when most needed. These results will be submitted to the Ontario Provincial Apiarist to update the treatment recommendations available to beekeepers. 
C. Best Management Strategies for Honey Bee Breeding and Colony Health
1. Colony Nutrition Study (CAAP)
The 2013 nutrition trails used the results from 2012 to improve the application of pollen substitute on colonies going to pollination of blueberries and blueberries & cranberries. It was identified in 2012 that there could be an issue with the quality, quantity and timing of pollen substitute application as it has been used by beekeepers to manage malnutrition. 
2013 controlled trials included three types of pollen substitute applied at three different rates and multiple applications when colonies were used for both blueberry and cranberry pollination. Three commercial beekeeping operations allowed the TTP to use 270 colonies for this trial. Results from this trial showed that there was a significant improvement using a pollen substitute recipe that is commonly prepared and used in the U.S. by beekeepers providing extensive pollination services. Not only did this substitute show an increase in honey bee populations, but brood production was also sustained. These results will give Ontario beekeepers that send approximately 25,000 colonies into blueberry and cranberry pollination, the tools necessary to improve the nutritional conditions colonies returning from pollination, it will also reduce the cost of applying pollen substitute by approximately 25-30% using the top substitute tested in this project. We believe this cost will be further reduce when commercial beekeepers are able to source the ingredients in larger quantities to supply their needs compared to the prices for the amounts purchase for this project.
2. Patterns of infection, economic damage, and potential novel control of Nosema ceranae in honey bee colonies (CAAP)
This project was aimed at determining the infection patterns and relationship of the fungus Nosema ceranae with honey bee mortality and colony development. An estimate of the economic damage of this new parasitic fungus can be obtained with analytical procedures that were developed, and new, natural medicaments, aimed at boosting the immune system of bees were tested. Nosema ceranae has been recently reported in Canada, thus becoming a new emergent issue for the beekeeping industry. We do not know much about the seasonal patterns of infection, infectivity, pathogenicity and economic damage of N. ceranae in Canada. Without this knowledge it is not possible to assess the risks that this emergent pathogen possess to the beekeeping industry and will not be possible to design effective control strategies against this fungus. The following were accomplished with this project: 
  • The PCR technique to identify nosema species was improved by incorporating a honey bee gene in a triplex reaction to insure that negative or positive results are accurate, by always detecting the bee gene. 
  • The fluorescence staining protocol to assess nosema viability was improved by adding extra steps in the spore purification and staining processes. Experiments were conducted to validate the repeatability and accuracy of assessing spore viability rates with fresh spores and inoculated bees. Results showed high repeatability (>90%) and a correlation between pre and post inoculation levels of spore viability. Therefore we concluded that the method reliably assesses the viability and infectivity of the spores. 
  • The nosema species, spore viability and infection levels of bee samples was determined for spring, summer and fall of 2012, as well as for spring and summer of 2013. Results showed that N. ceranae is the most prevalent nosema species of honey bees in Ontario and that nosema spore viability and infection levels, as well as the proportion of infected bees are significantly higher during spring than in other seasons of the year. 
  • The sampled colonies were assessed for population strength and food reserves. Results showed that low levels of nosema infection do not seem to have a major negative effect on colony conditions, but very high levels (1.3-3 million spores per bee) significantly reduce bee populations and food reserves of colonies. 
  • Experiments on survivorship of individual bees, with known infection levels, were conducted in the lab and it was found that N. ceranae reduces the length of life of bees by 14%. Experiments on bee survivorship in field colonies were also conducted and again, it was found that nosema infections significantly reduce the life span of bees when infections are high. Infected bees lived 25% less than healthy bees. 
  • A literature review on the effect of nosema disease on bees was conducted and a preliminary economic analysis was done with data of this project, value of hive products and information from the review. An equation was developed with this information to estimate economic losses caused by nosema infections of honey bee colonies. The analysis revealed that infection levels of >750,000 spores per bee could cause a higher economic loss per hive than the cost of its treatment. 
  • Experiments to test the effect of natural compounds on N. ceranae infection levels and on the immune system of honey bees were conducted. Results showed that out of 13 natural substances tested, 3 reduced nosema infection levels by more than 50% and warrant future research. The immune gene hymenoptaecin was also induced and over expressed, which indicates that these substances activate the immune system of bees, which may favour their natural defence against nosema infections. The results of this project are significant for Ontario and Canadian beekeepers because the techniques improved to diagnose nosema species and assess their viability can now be used by the industry to determine the severity of nosema infections in their colonies. Additionally, with the information generated in this study, they will be informed that nosema infection levels have to be maintained low (below 750,000 spores/bee) to prevent damage from the disease. The most critical season to prevent nosema disease damage is the spring because that is the time of the year when spore viability and infection levels are higher, particularly if the spring is cool and rainy (these conditions seem to favor the increase of nosema infection levels as occurred in the spring of 2013). It can also be recommended to beekeepers to monitor nosema infection levels in early spring to decide whether a chemical treatment is warranted. If an antibiotic treatment (fumagillin) is chosen, early spring is probably the most effective time to use it, rather than during fall when colonies are currently and regularly treated, probably with little effect on infection levels and causing a considerable expenditure to beekeepers; our data supports the notion that fall treatments against nosema, seem unwarranted. Furthermore, this project allowed the identification of 3 natural compounds that could have promise for future control of Nosema disease without causing resistance in the pathogen or leaving toxic residues in honey. The information generated in this project will be delivered to beekeeping organizations through seminars, talks, workshops and articles in beekeeping journals. The objectives of this study were successfully completed thanks to the collaboration of the Ontario Beekeepers’ Association and personnel of the University of Guelph. 

D. Progressive Training and Information Program for Beekeepers:
1. Introductory Beekeeping Workshops (TTP)
“Introductory Beekeeping” workshops were held in Guelph (May), Moose Creek (May), Ancaster (May), Burgessville (June), Shelter Valley (June) and Bracebridge (June). A total of 148 people attended these 6 workshops in 2013.
2. IPM and Beekeeping Workshops (TTP)
“Beekeeping and Integrated Pest Management (IPM)” workshops were conducted in Moose Creek (May), Guelph (May), Ancaster (June), Bracebridge (June). A total of 77 people attended these four workshops in 2013.
3. Introductory Queen Rearing Workshop (TTP)
One “Introductory Queen Rearing” workshop was conducted in Burgessville in June. Twenty-three people attended this workshop in 2013. 
All three workshops consisted of classroom presentations accompanied by hands-on sessions in the bee yard.
4. Advisory and Outreach for Apiculture in Ontario (KTT, in conjunction with U of G)
This project is designed to disseminate information discovered, obtained and translated by the University of Guelph, OMAFRA and the Ontario Beekeepers Association to all of the beekeepers in Ontario. Due to the large geographical distances between the scientific research taking place in Guelph and the beekeepers in all areas of Ontario, the ability to provide real-time current information to the beekeeping industry is difficult. Beekeeping is a dynamic, ever-changing industry, and the need to stay informed is important, so that beekeepers can sustain their operations and the health of honey bees is improved and maintained. By providing alternative means of attaining advisory materials, beekeepers living in remote areas are more likely to be informed.
This project involved going above and beyond what is generally provided as outreach for the beekeeping industry by using current media formats and new technology to distribute current information. The relay of up-to-date information through the use of INFOSheets, webcasts, recorded meetings, instructional DVDs and updated manuals assists in the success of the beekeeping industry in Ontario. INFOSheets provide guidance regarding disease and pest identification and treatment/control. Research is presented at yearly meetings that take place in various parts of the province and typically involves information that is not available in print format. Webcasts are the live-streaming of these meetings and provides an opportunity for those unable to attend, to interact with guest speakers and presenters. By recording the meetings, the ability to watch the presentations at a later date is achieved and it also provides a historical bank of information that can be assessed for current and future research. Instructional DVDs provide a visual, hands-on approach to learning how to become a beekeeper and maintain honey bee colonies efficiently. Updated manuals for Ontario-specific situations provide ideal instruction for local beekeepers. INFOSheets are developed, in both English and French, and distributed to the beekeeping industry at meetings, online, by email, by mail and through personal contact. As well, a Vimeo channel was created and each month, a selected speaker from these meetings is posted online for OBA members to view free-of-charge.
The instructional DVDs are being finalized with filmed video from the 2012 & 2013 beekeeping seasons. An Introductory Beekeeping DVD, and Integrated Pest Management DVD and an Introductory Queen Rearing DVD will be produced to accompany the manuals produced by the OBA. The Ontario Beekeeping/IPM Manual and the Ontario Queen Rearing Manual are updated and reformatted. New pests, diseases and current research in the industry, out-of-date information has been removed or updated and new graphic designs are incorporated by working with technical writer.