Introduction

As in the first Corona year, also in 2021, the participating beekeepers have been able to actively work in the program with spectacularly good results. We now see that the enormous amount of work done since 2014 proves that the assumptions we started with – that there is a little bit of Varroa Sensitive Hygiene (VSH) in the honeybee population and that that can be selected for by Single Drone Insemination (based on the work from the USDA going back to 1997) – are working out in practice.  

In this introduction we first give you a “refresher” on the method we are using, followed by the latest results.

The method is based on 4 types of mating of the queens; Single Drone Insemination, Multi Drone Insemination, Land mating stations and Island mating stations. Each of these types of mating has its own advantages (and disadvantages), but are powerful if used for the right purpose. That is why the overall program is a mix of these 4 types.

1. Single Drone Insemination (SDI):

After adding extra Varroa mites, we investigate the brood and determine the level of resistance (Varroa Sensitive Hygiene, VSH). This allows us to basically find those queens/colonies that were mated with a highly resistant drone. Only colonies with queens that remove 75% or more of the mites are kept. The daughters of these queens will carry the resistance genes, creating the basis of the breeding program.

Please note that at the beginning of this selection process we estimated that only 10% of the drones would carry enough VSH genes.

In the case you use more than one drone at this stage, like in normal mating with approximately 10 drones, the effect of one drone carrying enough VSH genes is diluted by the 9 other drones carrying no or little VSH gens. When VSH genes are diluted, there is not enough resistance left to detect/measure it.

2. Multi Drone Insemination (MDI):

The SDI colonies are very well suited starting from a population with limited VSH genes. SDI colonies can only be small and are relatively short lived (because of the limited amount of sperm stored by the queen). As they are small, the colonies cannot be easily selected for the other important traits as honey production and low tendency to swarm. For this reason, we inseminate a larger number of queens with the semen of at least 10 drones. As we are now later in the selection process, most of the drones will carry the resistant genes. These colonies can be kept much longer (2 to 4 years) and can be tested for the described traits. We also monitor the varroa population growth in these colonies by taking samples (300 bees per colony) to determine the number of mites on these bees (instead of in the brood), called the “phoretic” mite count. This is a method that can be used at large number of colonies without too much labor involved.

The advantage of using Multi Drone Inseminations is that we can keep the biodiversity as high as possible. We can use a large variety of different drone lines compared to a mating station that usually has only one or two different lines per year.

3. Land Mating station:

The disadvantage of the MDI colonies is that it requires special insemination skills. The amount of work involved makes it less suitable to do large scale production of mated queens in the broader beekeeper community. Therefor we are more and more making use of land mating stations. On such a mating station, ideally somewhat isolated from other apiaries, we place colonies that will deliver drones carrying varroa resistance genes. As this is a land mating station, we assume that a certain, hopefully low, percentage of the drones comes from colonies from other beekeepers. This means that the mated queens will not produce 100% “pure” colonies. However, the queens mated on these stations can be used in production colonies in which they will be assessed for our selection program.

So from these land mated station queens/colonies we do not produce further queen offspring (they could have low resistant genes from the not-pure drones), but we can definitely use the (pure!) drones for our selection program.

Indeed, the best colonies can then deliver well selected drones, because drones are coming from unfertilized eggs. They just represent the genes of their mother, they have no direct biological father!

We have experienced in our Hawaii program and now also in our EU program that using this step is actually very powerful. Given the relatively large number of colonies that can be produced and monitored (only a few samples of bee washing are needed for phoretic mite level assessments per year), a both strong and still broad selection can be made based on high resistance as well as good honey collectors, gentleness and with low swarming tendency.

4. Isolated mating station:

Whereas the previous methods are very helpful during the selection process, they are not very useful for diffusing the breeding progress to the beekeepers that do not participate in the selection program. Those beekeepers are interested in purely mated queens (“F0”) that  can be used to graft from in order to produce their own F1 queens for their production colonies. For this distribution of good traits, using an isolated mating station (an island or a remote mountain valley) can be a very efficient way. We will see that more and more isolated mating stations are using Arista or (at least partially) Arista derived queens as the basis for their drone lines. This will facilitate the distribution of the resistant genes in the different honeybee  populations – and will eventually even benefit beekeepers that are not visiting mating stations at all as their queens will be more often mated with varroa resistant drones from their neighbors.

It is important to note that we are not yet in a position to supply all these mating stations with new, non-related and highly resistant queens, year after year. We should also ensure that we do not lose the more than 100 years of selection (either by beekeepers or by nature) by moving too fast with the still limited number of Arista validated lines. We are now at the turning  point; we need the full support of the beekeeping community but also from the governments to extend the program and to ensure that we keep the genetic base of the honeybee as broad as possible. The method works, it is “just” a matter of implementing it 😊…

Progress in the breeding program

Organization

Dr. Guillaume Misslin, our Project Leader located in the Netherlands and Sacha d’Hoop, our Project Leader located in Belgium, support around 200 beekeepers distributed in 18 groups within: Belgium, Germany, France, Luxembourg, the Netherlands, Austria and Switzerland. In the 18 groups all three main races of bees are represented: Black, Carnica and Buckfast bees.

In Belgium we could welcome a new Project Leader: Julien Duwez. Freshly graduated in bioengineering from Gembloux, he was already involved in the project as a participant before starting to work with us. His arrival in the team will allow us to improve the coordination and the follow-up of the participating Wallonia beekeepers as well as to improve the training of those that recently joined us.

The beekeepers in the program are supported by the Project Leaders in the choices to be made in the use of lines, in the queen and drone breeding techniques and in the varroa infestation and counting sessions. Furthermore, the Project Leaders also organize insemination courses with the groups, so they gain more autonomy and flexibility.

The total number of colonies that has to be followed and monitored is increasing as we now maintain more and more colonies mated in the different described ways. For this we fortunately have our in-house, tailormade web-based database application “Queenbase”. It is more than ever the tool to be used; to report back to the Project Leaders the production and phoretic data of the colonies gathered during each season as well as the pedigrees and offspring data for the queens in these colonies.

Single Drone Insemination results

The number of colonies we could test for removal behavior (Varroa Sensitive Hygiene; VSH) in SDI colonies was 969, an increase of 56 compared to the previous year. As in previous years, about 1/3 of the colonies removed more than 75% of the reproductive mites (the minimum required to not need chemical treatment). These high VSH colonies can be used in the breeding program.

More than 100 of the high-VSH colonies removed all – 100% – of the reproductive mites from the worker brood. We have now demonstrated this year after year, but it is still a small wonder to see that the bees remove the Varroa mites so well themselves and that this trait is transferred into the next generation. To evaluate the colonies, volunteers open the brood cells to investigate its content. The total amount of opened brood cells is still increasing compared to previous years as at higher VSH levels we find less mites, so we open more cells to still get enough information. Furthermore, we do more and more recounts, just to be sure that the high scores are indeed sustainable if measured multiple times.

The fact that 2/3 of the hives are not high VSH is because we ask the beekeepers to keep bringing in new blood (good, new lines, but not VSH). Therefore, the program is a mix of new lines that are in the first years of selection (initially low VSH) and already proven lines (high VSH).

Land mating stations and Multi Drone Insemination results

Italy

As we described in the “Land mating” paragraph, and as we learned in our Hawaii program, you can make a very good contribution in the selection by using production colonies.

As also described in previous updates, we have been able to build a “well-oiled machine” with a group of highly skilled professional beekeepers in Italy. Every spring, and thus also in 2021, we delivered a set of SDI colonies to this group. From these, more than 300 queens are bred and mated with drones from colonies also coming from our program (selection from the previous year). These 300+ queens are heading production colonies that have to prove themselves in production and are followed and evaluated for several years on the phoretic mite infestation (simple sample of bees “washed”), gentleness, honey yield and low inclination to swarm. From the best colonies (after 1, 2 or 3 years) large amounts of semen are collected by capable inseminators/beekeepers during a second visit in the spring. This semen is transported in proper conditions and used for multiple-drone inseminations of breeder queens at the beekeepers from the different participating countries.

2021 is a special year for this selection. After all the years of preparation and pre-selection, this was the year that queens, inseminated in 2020, had to prove themselves (for more than) a full year. For this purpose, in 2021, 16 queens in the Netherlands were followed by taking samples. Throughout the full year these colonies, of course without any treatment, kept the mite infestation around half a percent! Control colonies (non-VSH) would have grown the mite levels to around 10% in the same timeframe.  We are very happy and proud of these excellent results – made possible by 8 years of work with a good cooperating group of now 200 beekeepers.

It is furthermore striking that already in spring these colonies seem much larger and healthier than “before” (in the early 1980’s, the Varroa mite entered our countries). And also in the fall the colonies are large and look more healthy compared to what we are used to.

Maybe this should not be a surprise? The bees are doing something that beekeepers cannot do that is removing Varroa mites 24 hours a day, 7 days a week. As a result, the pathogenic virus infections (distributed by the mites) never have a chance. This is the main difference with the current practice of chemical treatment. You can treat as good as you can as a beekeeper (e.g. 3 times a year), but the virus infections only decrease slowly after a Varroa treatment (takes weeks/months). In fact, you don’t directly treat against the viruses that are causing the main problem for the colonies. And after a chemical treatment against the Varroa, it just starts growing again, until the next treatment. The still present viruses can then just “hitchhike” again. And last but not least, there is no impact of chemical treatments on the bees. Now that we can actually start beekeeping with really healthy colonies again, we realize more and more to what we have become “accustomed” to in the past 40 years… it is truly a relief to be able to practice beekeeping with “normal”, healthy colonies again!

Belgium-Sélange

The method used with our Italian colleagues, is also implemented in Belgium and the Netherlands.

In Belgium we established the first transnational resistant mating station in Sélange (type “land mating station”). It is the result of a partnership with l’Abeille Arlonaise, a beekeeping section from the south of Belgium. Not only is the aim to allow high resistance matings, but also to create an area without varroa treatment on the long term. In 2021, the mating station was opened to members, generating around 360 queens from many different resistant lines, that can be used to produce males for future inseminated queens. It is asked not to treat any of the colonies that originate from those queens, unless a phoretic count proves it is necessary (which obviously eliminates these queens from the program).

Netherlands-Bronlaak

Also in the Netherlands we opened our own land mating station, close to our offices. This station is run by local beekeepers and coordinated by Arista. We have admitted 14 beekeepers for the time being. These beekeepers receive the starting material (larvae) for queen breeding from us and then have these virgin queens mated in small fertilization boxes. Just like in Italy, the fertilized queens can then be used in production colonies for the further selection of good drone material in the coming years.

Island mating stations

The VSH lines that become available have also drawn the interest of several island mating stations. They are reserving part of their capacity for putting lines on their mating station that have certain levels of VSH incorporated.  

Ameland (the Netherlands) has had several different VSH lines at the station in 2021 on a separate part of their island. This enabled Arista beekeepers to send their mating colonies and select for different lines.

Also in Germany there are more and more mating stations that can now include Varroa resistance as a selection criterion by using queens that have an origin in the Arista breeding program.

In 2021 a first start was made on a cooperation program with the Flevoland group/station. This group will from now on be solely dedicated with the selection and distribution of Varroa resistant queens. Already in 2023 the mating station will be “manned” with queens coming out of the Arista breeding program.

As earlier stated, we are too early in the program to fully switch to varroa resistant queens on all the mating stations. For that we need a more diverse and broader selection of resistant lines – respecting the large amount of work and quality available in the not-yet varroa resistant lines.

Research

Genetic marker for VSH

In the Genetic marker project we are trying to find the genes – or a genetic marker for these genes – that are linked to the VSH behavior (collaboration Arista, Van Hall Larenstein, Inholland, Bejo Zaden). We are very close to a first, very important analysis as full genome of the target colonies has been sequenced. If all goes well the first analysis on the full set of data will be available this summer (2022). Hopefully we will have a first indication whether we will indeed have a powerful tool to speed up the selection… to be continued!

VSH rapid test

It is important that we find ways to make the very labor-intensive selection more efficient and/or effective which is a reason for the described genetic marker research. The selection process could be more efficient if we would have a way to select for the VSH trait without having to breed, infest and count the Varroa mites themselves…

We are therefore pleased that the Goeie Grutten Foundation has granted us a subsidy (starting in 2022) to develop a VSH rapid test. Such a test would not require Varroa mites and would have to give a relatively quick idea of the level of Varroa resistance/VSH. For this, we will look at specific odorants that might be used in place of the mite. For this part of the research we work together with the University of Greensboro (US), the Buckfast Flanders group and the University of Leuven.

We will also investigate other ways to mimic the effects of the Varroa, without using the Varroa mites themselves.

Arista hives

At the end of 2021, we had a total of 21 installed Arista sponsor hives. These hives have queens from our selection program and are used to further evaluate and use the best lines. Both in 2021 and 2022 we already distributed drone brood and larvae from the best of these colonies to mating stations and participating Arista beekeepers.

A nice “side-effect” is that those (regional) beekeepers that take care of the hives often are new to our program. Their first-hand experience and their own network make our program known to a wider group of beekeepers.

Furthermore, we have spent a lot of time, with also highly skilled “pensionados”, on the proper functioning of the monitor system (weight/temperature/beecount in-out). The current version is very stable, works in all places where we have hives and is fully integrated with the software (Queenbase) of our breeding program.

The financial contribution from these hives is critical to the survival of our breeding operation. We hope we can further expand this program and welcome more donors/sponsors for an Arista hive!

Thank you

We thank our donors and sponsors for their financial contributions that enable us the organizing of this complex pan-European program.

We thank all the participating beekeepers and volunteers for the incredible amount of time, passion and expertise they invest in the Varroa Resistance breeding program on a daily basis.

Location & Covid

In the Netherlands, we moved from the family house in Katwijk to the ex-cow-AI-station in Beers in March 2020 (see our previous update). At this location we have a lot more space at our disposal and this actually saved us during the outbreak of the Covid pandemic.

In March and April, an instant decision had to be made: what to do with the breeding program? Not only the activities in Beers, but also the groups in all other countries (including Italy, close to an area with a major outbreak) were facing the question “what to do next”. 

Since a one-year pause would have been disastrous – after all, you have to breed a new generation every year – it was quickly decided to try and give it all to continue. That turned out very well. Even with social distancing, special logistics for inseminations and counting, and electronic meetings, we managed to achieve a “record” year. 

At the Beers site, we stayed open – with extra measures – and even welcomed extra volunteers.  We were able to do this without any transmission of Covid at work.

Progress of the breeding program

In 2020, the number of participating beekeepers in the various countries grew to 133.

This is also reflected in the number of tested colonies. In 2020, 912 colonies were tested for mite-removal behavior (Varroa Sensitive Hygiene; VSH) – a record number in the Covid affected year of 2020!

One third of these colonies (312) were found to be high VSH, removing 75% or more of the (previously added) mites from the brood. Colonies with 75% or higher VSH are able to survive without any chemical treatment. Of the 912 colonies, 79 colonies removed all mites from the worker brood, so these colonies are 100% VSH.

The best colonies are used for further breeding, also taking into account other characteristics, such as gentleness and honey yield.

As mentioned in our previous update, we have established a partnership with experienced (professional) beekeepers in Italy, whom we again provided with new queens to breed from this spring. The daughters of these queens are mated with drones from our program in a mating station. These queens are used for production colonies and the best ones are selected, based on mite infestation, honey yield and gentleness, among other things.

This spring, we were once again able to collect semen from the best colonies in Italy (daughters of queens we had brought in 2019 and 2020), for the inseminations of more than 500 queens in Belgium and the Netherlands. These queens received the normal amount of semen (from about 10 drones) and will be introduced and tested in large colonies. The first results look good (low mite infestations, high VSH in tested colonies), however, we want to follow the colonies a bit longer before we draw our conclusions.

In several countries, beekeepers have been trained by Arista to perform inseminations themselves.

Our project leader Dr. Guillaume Misslin made his 5000 km trip to the various participating beekeeping groups in Germany, France, Austria and Switzerland. During this trip he gave insemination courses to 17 eager beekeepers and inseminated 990 queens for the program. Teaching more beekeepers to inseminate enables the breeding program to grow further.

Mating stations

Apart from the “Italy route”, there are also more and more mating stations with colonies from our breeding program. On the island of Ameland, there are, as we speak, many colonies from beekeepers in our program. Starting from VSH queens, several lines of drones have been prepared by the Ameland group. Furthermore, there are land-mating stations in Germany and Austria, our own land-mating station in Belgium is now well used and at this time, the first queens are being mated at our land-mating station in Bronlaak, near Beers. At the same time, there are also mating nucs at a new station of the Amstelland group.

At all these mating stations normal “multi-drone” matings are performed, which enables the use and testing of these queens in large colonies. Clearly, we are entering a new phase in our program. We will continue the single drone inseminations (small colonies) to find new lines with high VSH, but will be making more and more large colonies, to be able to select on the other traits (honey yield, gentleness, swarm-weariness etc.) as well.

Research

The collaboration of Arista Bee Research, Bejo Zaden and the Universities of Applied Sciences Inholland and Van Hall Larenstein has succeeded in sequencing the complete genome of 48 high and low VSH colonies. Furthermore, the VSH behavior was recorded extensively on video. We are now working hard on the analysis of this first important data set. Meanwhile, 100 additional colonies have been selected to have their genome analyzed. We hope to have the data from all the colonies at our disposal at the end of 2021. It will be very thrilling to see whether we can find the needle (a piece of DNA code that correlates with the VSH behavior) in the haystack (the entire bee genome) …

Arista hives

In 2020, we started delivering the first Arista Adoption Hives in the Netherlands. This specially developed hive (with solar panels, computer and sensors) contains a queen from our breeding program. Such a colony actively participates in the selection and spreads the genes for the Varroa resistance behavior to other colonies in the surroundings through the drones.

The sponsor can follow the progress of the hive via an app, which shows e.g., the weight and the number of bees flying in and out of the hive.

Despite the Covid pandemic, we have been able to place 13 hives. Governmental (municipality of Cuijk, municipality of Mill and Sint Hubert and the province of North Brabant) and business organizations (several Rabobank locations, ZLTO, Triple, Torn) as well as charitable foundations (Adessium, Elise Mathilde) have adopted a hive.

The Arista Adoption Hive concept is rolled out with the support of local beekeepers and beekeeping associations. The Varroa resistant material of the colonies in this program is also shared with these beekeepers. Therefore, the program is both a selection program and supports the distribution of Varroa resistant colonies in the beekeeping community.

The program has already been very successful this year. From the best colonies, queens have been made available to mating stations in Ameland, Flevoland and North Brabant.

Thank you

We thank all the participating beekeepers and volunteers for the incredible amount of time, passion and expertise they invest in the Varroa Resistance breeding program on a daily basis. We thank our donors and sponsors for their financial contributions that enable organizing of this complex pan-European program.

Location

Where previously cows were artificially inseminated, it is now the turn for queen bees… So it remains a AI-station 😉. At this new location we have 2 large offices at our disposal, a kitchen, an archive, a laboratory and a spacious storage. And of course, a large outdoor area for our bees. It is great to have enough space for all the projects of our staff, volunteers and students!

Method

At the start of the selection in a new group or breed, the emphasis is on using the “single drone insemination” principle (SDI). We make small colonies and do not use the normal sperm mixture of 8-10 drones but sperm from only 1 drone per queen.

This is a very important step. By using only 1 drone, all workers have the same mother and father. This makes it much easier to select, as you can see all the characteristics come to the fore to a greater extent. As soon as the hive is on its own bees, extra mites are added and 2 weeks later, the degree of infection in the brood and the ratio of non-reproducing versus reproducing mites is examined. Next, we only use colonies with a low level of mite infestation and a high percentage of non-reproducing mites. In these colonies, most of the mites that reproduce have been removed by bees – a sign that the workers (and therefore also the queen) carry the Varroa Sensitive Hygiene (VSH) trait. It is these colonies that are used to continue breeding.

Where at the start of the selection mainly these 1-drone colonies are used, in the next phase more normal (large) colonies are used with queens inseminated with 8-10 drones. With these large colonies (the second group) it is easier to select for other important characteristics such as honey production, gentleness and disinclination to swarm. In these colonies the growth of mites is monitored by determining the infection on the adult bees. The best colonies are used as a basis for the queens of the next generation.

The third important group of colonies in the selection program are the open-mated (production) colonies. In these colonies the infection on the adult bees is also determined after a season of “production” (and obviously no anti Varroa treatments). The best colonies, both in terms of mites and honey production, can then provide the drones to the first and second group already mentioned for the next generation of colonies.

Breeding program in the USA

Hawaii, as we know, is not only a beautiful holiday destination but also a “Varroa paradise”. The mite can propagate here all year round in the beehive and beekeepers have to treat against Varroa 4 times a year.

However, with the application of the system with the three groups of colonies, as described above, beautiful results have been achieved since 2015.

We have now reached a very high level of resistance in our inseminated breeding queens. This means that this group has not been treated for two years now. In these colonies the mite infestation on the bees remains on average below 1% throughout the season.

These good results enable us to continue with a simplified breeding program. This program will be further supported by the USDA in Baton Rouge.

However, as in Europe, a greater effort is now needed not only to secure the existing lines but also to make several other lines/sources/breeds resistant. We have therefore set up an American sister organization of Arista in Dallas, Texas, to support the selection and breeding of Varroa resistant bees on the mainland of the US beekeepers.

Breeding program in Europe: Buckfast, Carnica and Black bee

In the Netherlands, Belgium, Luxembourg, Germany, Austria, Switzerland, France, Spain and Italy a growing number of Buckfast, Carnica and Black bee colonies have been infected with mites in recent years and assessed for their degree of resistance. Mainly mini-plusses (small boxes) were used, of which the queen was inseminated with sperm from one drone. In the first year, 2014, there were almost 100 colonies and in the past year more than 800, made by 119 beekeepers.

262 of these colonies were “high VSH”. High VSH means that 75% or more of the reproducing/fertile mites are removed from the worker brood. One third of these “high VSH” colonies was as high as 100% VSH. This means that in these colonies, even after adding extra mites, no mites with offspring can be found in the worker brood. This is something to think about for a moment…: So it is actually possible to grow bees that take all the reproducing mites out of the worker brood.

Now that we have reached this level with small 1-drone colonies we will make more and more of our colonies with the usual 8-10 drones (MDI: Multi Drone Insemination) in the coming years. Artificial insemination will continue to be the best method to keep as much diversity in the population as possible. You can make sure that you use as many different sources as possible on the maternal as well as on the paternal side.

As described in the paragraph Method, there is a third group that is very important for further breeding – these are the open-mated colonies that can serve as drone suppliers. This year we started to work with major honey beekeepers in Italy to put our lines into practice and test them for honey production and mite growth. Last year they were able to breed queens from 16 Buckfast lines (from NL, BE, LUX) and have them mated in a VSH land mating station. These queens have been introduced into normal honey colonies. From the best colonies, if all goes well with the Corona, we will collect sperm next month in Italy to make the next generation of breeding queens in the Netherlands, Belgium and Luxembourg.

For the distribution of pure material to large groups of beekeepers, the mating station on the islands work best. Arrangements have now been made for this with the mating station on Ameland. Ameland has recently received highly resistant queens from us (bred in Belgium) and will turn them into drones (i.e. breeding daughters to produce drones) that can then be used by participants of the Arista program next year. These beekeepers will then be able to have their queens mated on the island.

But we are not there yet. We will have to “convert” an even larger part of our Buckfast, Carnica and Black bee stock. In fact, we delivered a nice “proof of concept”, showing that the setup works in principle. With the proven method, we can now pick up and execute the selection with many more beekeepers. This requires more project leaders in the Netherlands as well as in the other European countries and the US. In order to allow larger groups of beekeepers to participate, we are implementing a full training and education program: “Arista Academy”.

Genetic Marker project

In the genetic marker project excitement is growing. We have succeeded in collecting bee samples from a large number of both very low and very high VSH colonies.

The Inholland University of Applied Sciences has already isolated the DNA of half of the samples and we are now in the queue for the determination of the whole genome (outsourced to a specialized lab). After this, Bejo Seeds and Inholland can use modern software to search for the differences between low and high VSH colonies! Searching for pieces of DNA that can be predictive for whether or not VSH is present…

With Van Hall Larenstein we also managed to make a large number of infrared video recordings of infected frames in low, medium and high VSH colonies. For the first time, we are able to “watch” what is really happening in the colonies on a large scale. Van Hall Larenstein is currently working on the first analyses of this large mountain of visual data. To be continued!

Publication of scientific overview article

Jacques van Alphen and BartJan Fernhout have written an overview article on the development of Varroa resistance in honeybees: “Natural selection, selective breeding, and the evolution of resistance of honeybees (Apis mellifera) against Varroa”. This article was published in June in Zoological Letters!

The article can be requested via the following link: https://doi.org/10.1186/s40851-020-00158-4. In this article we discuss the natural development of Varroa resistance in certain regions (like Africa) and the need for breeding programs in Europe and America.

Thank you

The growth of the past years has only been possible through the contributions and efforts of the donors and many beekeepers in the various countries. We would like to thank everyone for all their support and continue to focus on our ultimate goal: Varroa-resistant bees for all beekeepers worldwide!

US breeding program

In Hawaii we had a quite turbulent year. At the start of the year we were very happy to present the breeding program to the Brandpunt-TV crew which made the effort to join us to Hawaii and Baton Rouge and made a nice overview of activities and progress.

As can be seen in the documentary (link on our website), their visit coincided with a large increase of activity of the Kilauea Volcano. Large flows of magma erupted from a large area, destroying more than 700 homes. David Thomas, the beekeeper that houses our program, had to remove a large amount of honey production colonies out of the active area. The colonies in our breeding program were fortunately not affected and during the summer, the magma flows came to a stop.

Originally, a normal colony on Hawaii had to be treated against Varroa around 4 times per year – as there is no winter, the mites can multiply in the brood all year round. Half of the colonies in the program are now treatment free, while the other half is down to, on average, a little bit more than 1 treatment per year. The average amount of mites per 100 bees has been steadily decreasing, to below 2% on average. This is very nice as this is below the to-be-treated-against-Varroa level of 3%.

2018 was a bad year from the weather point of view (yes, even on Hawaii): there was a record level of rain which hampered a normal honey flow. Fortunately, 2019 looks to become a more normal year, which will enable us to do more selection on the amount of honey a colony collects – also an important trait for a commercial honey bee.

EU breeding program: Buckfast, Carnica and Black Bee

More than one hundred beekeepers are now actively running the selection program in Austria, Belgium, Germany, Italy, France, Luxemburg, The Netherlands, Spain and Switzerland. Almost 700 colonies were prepared, infested with mites and assessed on their resistance level. 189 colonies were identified as highly resistant (≥50% of reproducing mites removed from the brood). Of these, a third of the colonies removed all of the mites and are considered 100% resistant.

To enable the distribution of the resistant stock to a larger number of beekeepers and also to produce production colonies of the participating beekeepers in our program, isolated mating stations have started to use highly resistant queens of our selection program as a basis for the production of their drone producing colonies (usually 20-40 colonies per station).

Beekeepers can bring their virgins to these stations where they are naturally mated with the highly resistant drones.

Especially the Bayern Buckfast group is already very active with the supply of queens to three stations: Karwendel, Leyhörn and Ammergebirge in Germany.

There is also a station running in Luxembourg, which was joined by a  newly complemented station in Belgium (Sélange-Arlon), just on the other side of the border.

After the Hawaii trip, the Brandpunt TV-crew were eager to make a second documentary of our activities in the Netherlands (see also the website). In this documentary Gerbert Kos and members of the Marken mating station are explaining what is entailed in the project and how the mating station of Marken works.

In this documentary the importance of the pollination tasks of honey bees was also explained by our partner Bejo Zaden in Warmenhuizen.

2018 was the first year for the Black Bee group in Belgium joining our program. A total of 34 colonies were counted for VSH and already 8 colonies were found to be highly resistant – a very nice score for a first year!

The large growth in participating beekeepers has been made possible by the recruitment of two new project leaders.

Guillaume Misslin, both a very experienced bee breeder and a trained molecular biologist, joined Arista in April (made possible by financial contributions from the Adessium and the Dioraphte foundations). As he already participated in our program he was quickly up to speed and is now supporting the groups in the German speaking areas (Germany, Switzerland, Austria) and the Carnica groups in the Netherlands and Luxembourg. Guillaume was also active in the Barbados project.

The second project leader we were able to recruit in 2018 is Sacha d’Hoop de Synghem. Sacha is a graduated Bioengineer and beekeeper. His recruitment is made possible by financial contributions of the Walloon Government to the newly created entity Arista Bee Research Belgium.  Sacha quickly took up the massive amount of organizational and training tasks which resulted in an even higher number of participating beekeepers in Belgium (>60).

Terschelling project

On the island of Terschelling a Black Bee project was started with the cooperation of all beekeepers on the island. Many of the colonies on Terschelling seem to be quite “black”, judging by their colour.

However, Buckfast bees and Carnica bees have also been held on the island for quite some time. So, we started a small project to see whether there are still enough relatively pure colonies to start a Varroa resistant Black Bee breeding program.

For this, Merel Rookmaker and Bart Lubbers, students of the Van Hall Larenstein University of Applied Sciences sampled all the colonies on the island and used the “fingerprint” of the wings of the worker bees to determine how “black” the bees are. From the 150 sampled colonies, 30 colonies were considered black enough (with 65-92% of the workers fully black and the remaining workers close to this) to keep. The queens of the other (less black) colonies will be replaced with daughters of these black queens during the coming year. This selection process will be repeated to further increase the purity. At the same time, we will start monitoring the mite-growth in the colonies to get an idea on the current level of resistance. This will enable us to prepare ourselves for the Varroa-resistance-selection-part of the project.

Genetic Marker project

2018 was the first year of the “RAAK” project, the consortium with Inholland University of Applied Sciences, Van Hall Larenstein University of Applied Sciences, the company Bejo Zaden BV and Arista Bee Research. The different partners each have their own expertise and tasks in the project.

Arista Bee Research is responsible for the creation and identification of colonies that have very low and very high levels of VSH. From each of these colonies, 50 bees are collected and stored for analysis. We were able to collect samples from more than 50 colonies with either very low or very high VSH.

This is enough to start the next phase of the project: the extraction and sequencing of DNA which Inholland and Bejo Zaden have now started to work on. Once sequencing has been done, we can start comparing the low and high VSH colonies looking for differences between the DNA sequences.

Van Hall Larenstein started the phenotyping of the behaviour which means that we will confirm that it is indeed VSH behaviour (removal of the infested pupae/mites from the brood) in the highly resistant colonies, by investigating captured video of bees on Varroa infested frames of brood. 

Barbados project

We were able to conclude all the needed measurements on 10 untreated colonies in Barbados. The results over the year are quite stable: relatively low mite levels on the bees as well in the brood, clearly showing high levels of resistance. The USDA is now investigating the bee-samples we took for virus levels and possible Africanization.

We will be compiling the data in a publication which will give a more detailed overview in the results and conclusions. The project has been made possible by financially contributions from the Bayer Bee Care centre.

Arista Academy

During the last few years we compiled a lot of knowledge and “do’s and don’ts” relating to our selection program. At the same time there are a lot of beekeepers that would like to start participating in our program or that would like to further develop their skills.

So, with the help of a new volunteer, Marjolein Bemelmans, we have started to build a comprehensive training program; “Arista Academy”. We will work hard to make the program available for participating beekeepers in the spring of 2020.

Arista Cloud

As we now have shown that the Arista selection method is working, it is time to further expand the breeding program.

In line with the increasing number of beekeepers, the number of project leaders, technicians and staff members will also have to grow. This also implies further investments in the establishment and furnishing of a European center with offices, lab, apiaries and training space as well as investments in equipment such as insemination sets, microscopes, incubators and freezers.

So, we are currently very busy for the launch of a new sponsorship program, tailored towards companies, governmental institutions and communities: the “Arista Cloud”.

By adopting one or more Arista hives with a Varroa resistant queen, one would become both a participant and a sponsor of the Varroa resistant breeding program.

Such a hive would be fully maintained by Arista & partners and will be monitored for (lack of) Varroa infestation, honey collection etc. Both Arista and the sponsor can follow the hive on an internet-dashboard to see how the hive is doing. The colonies in the hives contribute in two different ways to the program: Firstly, the drones from these colonies will distribute the Varroa resistance by mating with queens in the neighborhood of the hive (up to 10 km from the hive). Secondly, Arista will select the best queens and re-use them in the breeding program to produce the next generation.

We will further update you with more details on this Arista Cloud program once we have completed our logistic preparations. We are working hard to breed queens, prepare special designed computer monitored hives, etc. . Interested parties can already contact us for more information.

Please watch the documentary of Brandpunt+ (KRO-NCRV) of our Varroa Resistant breeding project on Hawaii (English subtitles).

This breeding project has been made possible by a cooperation of the USDA in Baton Rouge, Hawaii Island Honey Company, Project Apis m. and Arista Bee Research.

To help us to continue our projects, you can support us by making a donation.
Thank you very much for your interest & support!

https://brandpuntplus.kro-ncrv.nl/
https://www.ars.usda.gov/southeast-area/baton-rouge-la/honeybeelab/
https://www.projectapism.org/
https://aristabeeresearch.org/donors/

Boxmeer, May 17th, 2018.

Full resistance in the US Breeding program
In the US we have been working very hard to establish a breeding centre for Varroa resistant bees, in close cooperation with Bob Danka from the USDA in Baton Rouge (supplying pre-selected semen from the USDA bee-stock and financially supporting the program), Danielle Downey from the non-profit organization Project Apis m. (financial support and administration) and David Thomas from the Hawaii Island Honey Company (infrastructure, personnel and financial support).

A dedicated laboratory was built on the Big Island of Hawaii. On Hawaii we have a long season, enabling us to produce and select multiple generations per year.

With the financial support we have been able to recruit and train 4 employees, so since last year we are able to run the program “full throttle”. We have 200 small colonies for housing our inseminated queens and 600 large hives to house our breeders and drone supplying colonies. All these colonies are tracked with our in-house developed “Queenbase” software application.

In addition, David Thomas is running all his honey production colonies in Hawaii (several thousand) with queens from the breeding program. The progress made in the program is very encouraging and also gives us a lot of information which we use in the EU breeding program (and vice versa).

With commercial queens, colonies on Hawaii required treatment up to 4 times a year (brood in the colonies year round, so a “Varroa-paradise”), in contrast, the bees from our program are currently treated less than 1 time per year on average. The best lines reach full resistance and keep the mite infestation levels very low during a full year – without any treatment.

We are now working on establishing these high levels of resistance in a broader selection of our lines. Furthermore, we also have to ensure that the lines perform well from a honey harvest and pollination point of view and that they perform well in the commercial operations on the mainland of the United States (so field-trials are being performed).

The very good results on Hawaii show us that fully Varroa resistant bees are feasible and encourage us all to keep working hard – both in the US and in the EU.

Strong growth in our EU-breeding program
We started our first year – 2014 – with 6 breeders in our breeding program. The number of breeders has grown since then from 16 in 2015, to 35 in 2016 and up to 66 in 2017! In Europe, we currently have participating groups in Austria, Belgium, Germany, Italy, France, Luxemburg, The Netherlands, Spain and Switzerland.

With the growing number of breeders and breeding groups, the number of colonies being tested in the program increased to 390 in 2016 and grew further to 646 in 2017. 134 of these colonies showed high resistance against Varroa (> 75% of the reproducing mites are removed from the brood). Colonies with this level of resistance do not need treatments against Varroa. Twelve colonies did not have any reproducing mites at all in their brood (after receiving 100 to 200 mites two weeks earlier and then checking 300-400 brood cells) and are considered fully Varroa resistant (100% VSH). As these are in most cases one-time measurements, we will perform additional measurements on these colonies and use them in 2018, to confirm their level of resistance. The queens of these highly resistant colonies are usually inseminated with just a single drone. These single drone inseminated queens allow us to make quick progress in our breeding program, but they can only be housed in small hives and have a limited life expectancy, because of the limited amount of semen they received. We are now in a position where we can also start to produce more multi (8-12) drone inseminated queens, using the material from our resistant single drone colonies. These multi drone inseminated queens can be housed in regular sized hives and we will start evaluating these queens on other important traits, like honey production and winter-survival.

We will continue to look for extra financial resources so we can further increase the number of beekeepers and colonies in the program in order to obtain the needed high diversity in the resistant pool of colonies.

Carnica, Buckfast and now also Black Bees
We started our program with Buckfast and Carnica beekeepers. These two races represent a large proportion of the honeybees in Europe and so the majority of our current program consists of different lines from these races.

However, there are local native sub-species in Europe that are considered important, given their unique set of traits and genetic background. One of these races is the Black Bee (Apis mellifera mellifera) which is one of the ten recognized European bee sub-species. With the popularity of the Buckfast and Carnica, and the extinction of feral, natural wild colonies due to Varroa, several of the Black Bee populations are considered endangered and could also benefit from a Varroa resistant breeding program. So in 2018 Arista will start working with the Belgian Black Bee group Mellifica.be in creating a first set of single drone inseminated queens, which will be tested and selected for using our established method.

Furthermore, the beekeepers on the island of Terschelling (the Netherlands) have joined up with Arista to create a Black Bee breeding program on the island, making the island a reserve for Black Bees. This breeding program will also include the selection of Varroa resistant lines, using our selection method. In 2018 we will find out how pure, how “black” the bees are on the island. Based on the findings, we will make a plan to establish a pure, Varroa resistant, Black Bee population.

Genetic Marker projects
As we now get a first base of fully resistant colonies, it becomes possible to start projects based on these bees. The standard method which we use to select for Varroa resistant colonies is quite labour intensive. It would be a big help if we could test and screen our bees with a simple (genetic) test – if only such a test would exist…

To develop such a test, it is necessary that we improve our understanding of the underlying genetics of the Varroa resistant behaviours, like Varroa Sensitive Hygiene (VSH: the detection and removal of the mites from the brood). For this reason Arista formed a consortium with the Inholland University of Applied Sciences (Amsterdam) and Van Hall Larenstein University of Applied Sciences (Leeuwarden, Velp) and the company Bejo Zaden BV. Last summer, together, we managed to get a “RAAK-pro” project proposal approved and we have now started to work towards finding a genetic marker for the resistant behaviour. Arista will supply the bees for the project and will support Van Hall Larenstein with analysing the behaviour (“phenotyping”) of the bees. Inholland and Bejo Zaden will look for the genetic background of the behaviour (“genotyping”) by comparing bees that do and do not perform the specific behaviour. This project is supported by a committee composed of members of the NBV (Dutch Beekeepers Organization), BBV (Buckfast Beekeepers Organization), VCI (Carnica Beekeepers organization), the Laboratory of Genetics from the Wageningen University and the BVNI (Professional Beekeepers organization).

Arista has also started to support the BeeStrong project of the INRA institute in Avignon, France. 40 Arista colonies were counted last summer for VSH and bee samples were taken for genetic analysis at INRA.

Barbados project
There are a few places in the world where it is believed that honeybees have become resistant against Varroa, without the help from humans. Barbados may be one of these places. When Varroa established itself on the island in 2002/2003, the bee population crashed and most of colonies collapsed. Most of the colonies on Barbados are “feral”, so unmanaged colonies, living in the rainforest. A small number of beekeepers catch swarms of these feral colonies to populate their hives. Most of these managed colonies also collapsed, because beekeepers did not treat against Varroa.

Interestingly, after several years the beekeepers again started to get phone calls, asking them to come remove swarms. Now, around 15 years after the first outbreak of Varroa, the population is considered to be fully recovered and beekeepers catch swarms and collect honey from their colonies, just like before the Varroa reached the island.

With the financial support of Bayer, Arista is now establishing a small project on the island. This project has the aim of establishing and following an apiary with colonies for one season and regularly determine mite levels on the bees and in the brood to establish the level of Varroa resistance. Furthermore, we will try to get a first indication which trait (possibly VSH?) the bees are using to keep Varroa at bay. In addition we will check the haplotypes of the bees, to exclude the possibility that the bees now present on Barbados are immigrated Africanized bees.

At the last visit, 10 colonies (untreated) were investigated on mite levels. The colonies showed very low mite infestation levels in the brood and most of the colonies also had low levels of Varroa infestation on the bees. So first data indicate indeed high levels of resistance. Will be continued…

Personnel & organization
In addition to the financial support that was established for the genetic marker, Barbados and US-Hawaii projects, both the Adessium foundation and the Dioraphte foundation have started to financially support our breeding program! These are important contributions as we can now build a small organization that can support the quickly growing group of beekeepers in our program.

The combined funding enabled us to have BartJan Fernhout move from the Board to the position of program director and to recruit Guillaume Misslin as a project leader. In addition, the Van Hall Larenstein (VHL) University of Applied Sciences is recruiting a project technician for the RAAK-pro project, which will be working with VHL students at the Arista location.

Furthermore, the Wallonia government in Belgium has approved the funding of a project leader for Arista! To achieve this, we founded a legal entity “Arista Bee Research Belgium”. This enabled us to recruit Sacha d’Hoop de Synghem who will be dedicated to work in the Wallonia area while working closely together with the Arista team in the Netherlands.

And last but not least we have been able to rent part of a house with a large garden, giving us space for an office-lab and an apiary.

We are very excited that we can now really start pushing towards our goal: Varroa resistant bees that thrive and keep themselves healthy, without relying on chemical treatments. For this we will further work on expanding the donor base, the professional support and the number of participating beekeepers and beekeeping organizations. The ultimate goal is that all beekeepers in the US and the EU can stop all chemical treatments against Varroa, while strongly improving the health of their colonies and strongly reducing the number of colony losses. This would ensure an uninterrupted supply of honeybees supporting the agricultural industry with pollination services and honey production. It would also support the recovery of the native local European subspecies of honeybees and the re-establishment of natural populations.

Arberg, November 2016.

New Buckfast group in Bavaria (Germany) got off to a flying start.

In Bavaria (South of Germany) an already very active Buckfast-Beekeeper and Breeder group (having grown from 100 to 500 members during the last 5 years – buckfast-bayern.de) has joined the other Arista VSH Buckfast and Carnica groups in the search for Varroa resistant lines.

Josef Koller, the chairman of this group, has been working on breeding Varroa resistant bees for 20 years. In recent years he has been supported by a growing number of interested people (one of them being Ralf Höling). Through his concept, he succeeded in selecting colonies that survived untreated for years. However, so far it had not been possible to make offspring inherit this trait consistently.

So, inspired by the stories from the already established Arista members, it was decided to start a Single Drone Insemination VSH project. Many emails were sent during the winter and so the group started very well prepared in the spring of 2016. More than 40 test colonies were created, using mainly two Buckfast lines that had proven to have the lowest mite counts during the previous years.

The  large experience in the group with Varroa counting (determining infestation levels), combined with the Arista experience paid off as large amounts of mites were harvested, with newly constructed buckets and many kilograms of powdered sugar, to infest the 40 test colonies.

Early August the group (combination of beekeepers, family, friends and Arista volunteers) came together for three days and established the level of VSH for each colony by counting the reproducing and non-reproducing Varroa’s in the brood.

It became clear early on that all the years with few or no treatments whatsoever (with the consequential colony losses) paid off, as in their first year, half of the tested colonies already expressed high levels of VSH. With these good results in mind, the group immediately took action and organized another insemination session in the fall to produce offspring from the colonies that were found to have the highest VSH level. 93 colonies were created and these colonies will be available in the coming years to produce drones for new Buckfast combinations.

After this very good start, the group in Bavaria with coordinator Stefan Luff, as well as the neighboring Buckfast-Süd group, are preparing themselves during this winter for the coming year, to expand the number of colonies and lines in the program.