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SmartBees Newsletter 3, December 2016


SMARTBEES is currently about halfway through the four-year duration of the project. In this newsletter, we present highlights of our work so far, with special emphasis on topics of most direct relevance for beekeepers and bee breeders.

The whole project team of SMARTBEES wishes you a joyful holiday season and all the best for the coming year.
Resistance against Varroa and deformed wing virus (DWV)

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One of Smartbees’ objectives is to identify individual honeybees exhibiting Varroa resistance behaviour and mite reproduction suppression in order to identify genes and genetic variants associated with the observed resistance phenotypes. More than 44 000 bees from 3 subspecies have been individually tagged and screened for varroa-sensitive behavior. In this process, we have identified hygienic bees (those initiating opening of Varroa infested cells and those that continue these cell openings). These groups as well as a control group have been genotyped and we are analyzing them to find significant differences in their genome.

The severity of viral diseases related to Varroa infestation can be surprisingly different from apiary to apiary. To study the causes for this variation we have established methods to assess the susceptibility of different honeybee genotypes to deformed wing virus (DWV) infection. We have applied this method to 23 colonies and we have identified colonies that we will study to find genetic reasons for this difference in DWV tolerance.
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Moreover, Smartbees aims to increase our understanding of honeybee resistance towards DWV, which is vectored by Varroa and thought to be responsible for an important part of Varroa-related damage to colonies. We are determining how the immune system of bees interacts with the virus and whether honeybee resistance to DWV can be influenced through external stressors or changes to the diet of bees.
 
 Honeybees screened for their ability to detect and open brood cells infested by Varroa mites
 So far, we have collected and purified DWV from naturally infected bees (with crippled wings).
We have injected 2500 larval bees with different dosages of this DWV and compared their survival rate and presence of crippled wings at emergence with a control group of larvae injected with a similar amount of buffer solution. We have extracted RNA from the emerging bees to examine the level of DWV infection.
Varroa and DWV - a symbiotic relationship
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Another issue we have been studying is the role of salivary factors for the relationship between Varroa mites and DWV. DWV replicates (i.e. reproduces by making new copies of the virus) both in honeybees and Varroa. When Varroa mites suck hemolymph from honeybees, DWV is injected along with saliva. In this way Varroa works as a vector for DWV. Relatives of the Varroa mite are known to manipulate the immune system of their hosts. To study the effects of Varroa saliva on the bee and on viral replication, we have developed a method to collect saliva from the tiny mites and have injected this saliva or DWV into bee larvae and pupae from a Varroa free colony with no/very low DWV. Now we are analyzing their immune response and DWV levels. We also developed a new method to preserve RNA in Varroa mites that are sent between laboratories in international collaboration, making such collaboration easier.

An investigation into the molecular basis of the interaction among the “bee – DWV – varroa“-system revealed that DWV disrupts the immune reactions that can interfere with hemolymph uptake and use by the mite. Thus, we discovered a mutualistic relationship favoring both Varroa and DWV – at the detriment of bees.

DWV and its ability to cause disease

There are many different variants (strains) of DWV. Smartbees is elucidating the diversity of DWV sequences, how the virus may be adapting to its vector (Varroa), and which consequences these adaptations may have for DWV virulence (its ability to cause disease). We have improved methods to detect and quantify DWV strains (variants), and we have started the collection of different DWV strains across Europe. Much of the work studying how DWV is adapting to Varroa is done in the laboratory where we infect cell lines with DWV. A cell line is a cell culture derived from one cell, which can proliferate in the laboratory. There are no cell lines of Varroa available, but there are many cell lines from related ticks and we found 8 where DWV did replicate. They will be used for further experiments to see how DWV strains change when DWV switches host from the honeybee to the Varroa mite..
High tech tool for bee breeders
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Another objective of Smartbees is to develop a low cost tool for breeders to rapidly detect subspecies affiliation and presence/absence of genetic resistance factors against Varroa and DWV. This will be helpful in the selection of colonies for further breeding. This tool is a DNA analysis, which contains DNA markers that are specific for the different European subspecies, markers for resistance against Varroa and DWV, and markers for CSD (sex determination gene). Currently we are in the process of identifying those sections of DNA that are most interesting in this context. When this tool is available, breeders can take a bee sample and send it to a laboratory for analysis, and receive a report on the subspecies affiliation and potential disease resistance of their stock. We have tested methods for sampling and shipment of samples that can be easily used by bee breeders. Squashing honeybees on a filter paper, drying the filter paper and sending it with ordinary mail proved to be a good method to sample honeybees and the DNA was preserved well enough for genotyping when arriving the laboratory.
Beebreed and breeding strategies
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Europe is the home of approximately 10 different subspecies and many more local strains of honeybees. It is becoming increasingly clear that locally adapted bees are often more robust to parasites and diseases, but they often lack behind commercial strains with regard to traits such as honey production or gentleness. Smartbees is creating tools for the breeding of resistant and diverse bees, based on local stock. The multi-language website Beebreed.eu is one of these tools, which allows local breeders to benefit from the use of modern quantitative genetics. We have organized online translation of the Beebreed.eu website and it is now available in English, German, French, Spanish, Italian, Romanian, Macedonian and Norwegian. The translation into Portuguese, Danish, Hungarian, Greek, and Russian is under way.
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Andrew Abrahams and his colonies of black bees on the Isle of Colonsay, Western Scotland
Smartbees will suggest optimal breeding strategies for each participating country/population. These suggested breeding strategies are based on simulations of breeding efforts in the respective countries. Through a survey for the national breeding coordinators we have gathered baseline information for the simulations, such as the structure of the organized breeding efforts under Smartbees support, quantitative data on beekeeping and breeding, and evaluations of the characteristics of the native bees in comparison with commercial subspecies used in the country. The aim is now to use the simulations to give well-founded advice on how to optimize factors like mating control, the number of drone producing colonies in mating stations, and the choice of breeding queens.
Characterizing European honeybee subspecies

In order to efficiently protect and valuate bee diversity, it is important to know what is left of it. Another objective of Smartbees is therefore to characterize the genetic and morphometric diversity of European honeybee subspecies. To accomplish this, we initiated sampling of all honeybee subspecies in Europe in 2015 and continued this activity in 2016. Each population is represented by 96 samples of bees, each one from a different apiary.
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Picture A

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Picture B

Honeybee subspecies can be characterized both by morphometric data (such as wing venation, hair length, picture A) and their genetics (picture B)

To date, more than 1500 samples from 916 beekeepers have been registered. The samples are now processed for whole genome sequencing that determines the complete DNA sequence of the bees. Until now, we have generated morphometric data (wing venation, color, hair length etc.) from subspecies samples from Greece, Russia, Ireland and Poland. The samples will also be used in the development of a low-cost tool (DNA analysis) that will allow breeders to determine the subspecies affiliation of their bees.
Collecting information from beekeepers in Europe

An important part of Smartbees is to promote the use of regionally adapted bees. To do this it is necessary to understand beekeepers’ motive for their choice of bees. We therefore designed a questionnaire to gather this information as well as to survey beekeepers’ and breeders’ knowledge needs and preferred extension methods. The questionnaire was translated into 13 languages and launched through the SMARTBEES web page to survey beekeepers across Europe.
A total of 6 250 beekeepers have answered the questionnaire, but the response rate achieved varied substantially between countries. A preliminary analysis of the answers obtained so far shows that the knowledge of the native bees in their area is poor among many beekeepers.
Further, the concept of subspecies is not well established among beekeepers, with regard to the value of conservation of local honey bee subspecies. We can conclude that large differences exist between beekeepers’ interests, actions and knowledge in the different European countries.
 We will use the results from the second part of the survey to suggest new extension tools and regionally adapted communication strategies for sustainable management of honeybee populations on a European level. It is important to adopt an efficient dissemination strategy for project results from Smartbees, but also for extension to the European beekeeping sector in general.
Smartbees aims to support existing conservation effort for the European honeybee subspecies and we have collected information on conservation efforts (conservation areas and associations to conserve a specific subspecies) via the Smartbees webpage.
 This survey is still open and we would appreciate it if beekeepers could help us complete our knowledge of conservation areas and other conservation efforts by answering this survey (entitled conservation network) on our website. We plan to organize a meeting for beekeepers and managers in the areas of conservation at the upcoming Apimondia conference in Turkey in 2017.
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We have also started to gather information on the presence of feral colonies (surviving in the wild without any beekeepers looking after them).

Participate in the survey...
Breeding and performance testing of European honeybee subspecies
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Smartbees recognizes the need for more systematic breeding to improve and conserve the European subspecies of honeybees.
Smartbees has therefore launched breeding programmes for all European subspecies, as the first steps towards the breeding of resistant, locally-adapted stock. In 2015 we initiated field testing and selection of local bee populations by 20 training events, in which a total of 320 beekeepers from 21 European countries participated. To achieve harmonized testing procedures we developed a protocol for performance testing including 12 step by step and visualized methods and tests for the assessment of common and Varroa related traits.

The protocols can be downloaded from www.smartbees.eu/extension. They are supported by a Smartphone application (www.testbees.eu). The performance testers received a Smartbees performance testing kit containing necessary tools and equipment for the performance testing. The following parameters are recorded in the tested colonies: colony development, defensive behavior, calmness on combs, swarming behavior, honey productivity, natural varroa mortality, varroa infestation level and hygienic behavior.
The interest from breeders and beekeepers has exceeded our expectations and in 2015 and 2016 a total of 128 testing stations with more than 1400 colonies from 9 subspecies were established. The 2 year performance testing process is in its final stage and the data from the first test generation will be available soon. However, for some of the populations and subspecies these are first ever data relevant for breeding purposes. The results from the performance testing will now be evaluated by calculation of breeding values via the translated webpage beebreed.eu that will be used to select colonies for further breeding.
Additionally, Smartbees partners established 130 testing colonies for assessment of the Varroa sensitive hygienic (VSH) traits in populations of particular interest. We found variable, but high levels of VSH traits in several populations that suggest that these traits can be improved further by selection.

Download the protocols for performance testing here...
Impressum / Legal note
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SMARTBEES is funded by the European Commission under its FP7 KBBE program 2013.1.3-02 Grant Agreement number 613960.