Queen Insemination in Germany

At the end of our recent cycling trip through the Balkans, Slovakia and Austria, we spent the last 5 days with our German beekeeper friends, Andreas Hahnle and Christiana Keppler, at Wallenstein. The morning after they picked us up from Frankfurt, Andreas had me out at 7.00am, helping him to catch and cage virgin queens from mating boxes in his back yard. Why catch the virgins if they were in mating boxes, you say? Because these virgins were destined to be instrumentally inseminated.

Each of their mating boxes was fitted with a queen excluder entrance to ‘lock’ the queens inside. Each of the 25 virgins caught was placed in a cage and returned to its mating box, and all 25 mating boxes were placed in the ‘troopie’ before we drove some 200km from Wallenstein to Würzburg Bee Institute, where the virgins were to be inseminated by Frau Christa Winkler. The whole process of catching and caging the queens, and indeed the whole day, was marked by the NON-USE of a smoker, evidence again of the gentle nature of the German honeybees.

While travelling to Würzburg, Andreas explained that the Winklers were from near Berlin, so had been in the old East Germany. Prior to the collapse of communism and the demolition of the Berlin Wall, Herr Winkler had sometimes had to queue for up to three hours, in order to obtain scraps of stainless steel to make the instruments for his wife to perform the insemination of the queens. They had also been unable to obtain gas regulators with which to control the release of the Carbon dioxide used to anaesthetise the queens, so had developed their techniques using ordinary balloons to control the gas flow. So successful were they in developing their low-tech techniques that they were still using the same instruments and techniques today. Indeed, so successful were they in developing their instruments, if you look at a catalogue of insemination instruments, some of the equipment has been named after them!

The Winklers are now retired and in their early 70’s, but are reluctant to let their hard-won skills be wasted, so each year they do a ‘road trip’ for two months around Germany and France, towing a caravan, and carrying with them all the equipment needed to inseminate queens. Along the way, they offer an insemination service for beekeepers and queen breeders to have virgins instrumentally inseminated – they normally spend two days at each stop, then a day or two travelling to the next destination, and at each stop they inseminate queens for any beekeeper willing to bring the queens to them for insemination. This year the road trip would also include Luxembourg and Belgium

There were only two catches to this offer:

1.       The queens to be inseminated had to have a pedigree, going back at least two generations, giving them status within the German/French breeding programme (aimed at gentility, Varroa-resistance etc)

2.       And sexually-mature drones with a similar pedigree have to be available at each destination. This is solved by co-ordinating the whole trip with the German Bee Institutes, who raise the drones and have them available at each stop.

The upshot of all these efforts is that the Winklers and the German Bee Breeding programme have a low-cost, co-ordinated mechanism whereby they can disseminate the required genetic pool around the country. Not only is each beekeeper given a signed certificate authenticating the parentage of each queen, but at the end of the road trip, Frau Winkler spends another month completing all the paperwork, so that the entire insemination work she has done is documented and added to the database, providing a computerised backup to support and extend the breeding programme (evaluation of all the queens inseminated is also carried out and documented, but that is another story entirely.)

On arrival at Würzburg, we unloaded Andreas’ queens, and learned that his queens would be done after the 30 or so lined up already, waiting in line (the queue is all arranged with appointment times organised in advance – we are talking about Germany, the country renowned for efficiency!)

Andreas’ queens in queue

Andreas’ queens in queue

Queens already inseminated, awaiting the trip home

Queens already inseminated, awaiting the trip home

All told, the Winklers were to inseminate 130 queens over the weekend, although they often do 75-80 queens each day at a stopover. While we awaited our turn, we chatted with both the Winklers and Dirk, the extension officer from the Institute, whose job it was to supply the drones. As we watched the Winklers working as a team, their long years of experience was evident, and their co-operation streamlining the process along the way.

When our turn came, we removed half the queens from their cages, placing them in a box which was then given to Herr Winker. The queens were then given a mild gassing with CO2and then allowed to recover – this ensured they were tested for suitability for the insemination process.

The opportunity to watch the whole process now proved to be an extremely enlightening experience, as Frau Winkler insisted I sit by her side and every now and then be given the opportunity to look down the field microscope as she carried out the procedure. As we sat side by side, she enquired after Gretchen Wheen, about whom she had heard much, but whom she had never met. It was an opportunity for me to explain Gretchen’s formation of the Wheen Foundation, and the respect shown by the Germans for the work done by Gretchen in Australia was inspiring(see article later this issue on the seminar to commemorate Gretchen’s contribution to beekeeping in Australia).

The carbon dioxide treatment is a very important factor to stimulate egg laying. Two treatments are necessary to initiate egg laying, one during the procedure and a second given one day before or one day after the insemination. A second treatment of only 3 to 5 minutes is sufficient. The interesting ‘sidelight’ to the Winkler’s procedure was that the first gassing in the cage with CO2meant that each queen was gassed three times, instead of the normal two, but Frau Winkler explained that she felt this was not a problem.

 The queens being gassed before insemination. After recovery each queen was released inside a glass-walled box, and before Frau Winkler would perform the insemination, the queen had to be observed climbing up the glass, to show her claws were undamaged – this lessened her chances of being rejected when returned to the hive and released.

 The queens being gassed before insemination. After recovery each queen was released inside a glass-walled box, and before Frau Winkler would perform the insemination, the queen had to be observed climbing up the glass, to show her claws were undamaged – this lessened her chances of being rejected when returned to the hive and released.

Each queen was placed into a cylindrical backup tube. When she reached the end of the tube with the small hole she backed up into the adjacent holding tube. The holding tube (with the queen) then fitted on the queen holder of the instrument, which was connected to the carbon dioxide line. The queen was ready for insemination when she was motionless from the anaesthetising effect of the CO2.

Frau Winkler at the microscope, Herr Winkler in the check shirt. CO2 bottles in the background. In the foreground is the glass-walled observation box to inspect the queens.

Frau Winkler at the microscope, Herr Winkler in the check shirt. CO2 bottles in the background. In the foreground is the glass-walled observation box to inspect the queens.

After performing the procedure on the first 12 of Andreas’ queens, Frau Winkler called a halt – she needed to extract sperm from more drones. These had to first be removed from the drone mother hives, whereupon Dirk revealed a neat method for catching the drones – these had been confined upstairs in their mother colonies the day before, and all that was needed was to place a drone cage abovea slide in the hive lid. When the slide was opened, the drones flew out into the cage and once the cage contained enough drones, the two slides (one in the drone mother lid, the other in the drone catching cage) were closed – it had not even been necessary to open the mother colony.

The cage was then returned to the lab, the drones released into the observation box, and one at a time the drones were caught, everted and the semen collected from the endophallus with a pipette. In all about 150 drones were used, some of the semen intended for the following stopover.

Drone catching, German style.

Drone catching, German style.

Everted drone endphallus, ready for semen collection with a pipette. The whole semen collection process was carried out so quickly that it was hard to follow all the manipulations that had been carried out.

Everted drone endphallus, ready for semen collection with a pipette. The whole semen collection process was carried out so quickly that it was hard to follow all the manipulations that had been carried out.

Once all the queens had been inseminated, Frau Winkler insisted we wait for at least 2 more hours before departing from home, so as to maximise the recovery of the queens. While we waited, Andreas paid the 14,50€ for each of the queen inseminations – about $19.00AUD per queen and Frau Winkler completed the certification, after which we loaded our precious cargo for the 200km journey home.

Three aspects of the exercise struck me:

1.       The sheer logistics of the whole exercise, with each successive venue having to have available the sexually mature drones, and with the whole process being documented and evaluated afterwards.

2.       The co-operation needed between the researchers and the beekeepers to ensure the even dissemination of the genetics developed by the Varroa-resistance breeding programme throughout the larger area of Northern Europe.

3.       The willingness of the Winklers to use their technical expertise for the betterment of the honeybee ‘industry’ in Europe. The price charged for the insemination of each queen was, after all, really only a pittance – but was affordable enough to ensure that a large number of queens were inseminated, thus ensuring the viability of the whole programme.

The next day, the gentle nature of the German bees was again demonstrated as I helped Andreas put out queen cells, with minimal use of smoke and protective clothing. The tower from the ruined castle, Schloss Wallenstein, is in the background.

The next day, the gentle nature of the German bees was again demonstrated as I helped Andreas put out queen cells, with minimal use of smoke and protective clothing. The tower from the ruined castle, Schloss Wallenstein, is in the background.

Author: Des Cannon