Artificial insemination of queen bees as well as breeding of bees with artificial honeycombs

Nicot systems

Dzhentersky

interlinear

interbreed

Heterosis, they are fertilized with micro doses (two to three microliters) of sperm from breeding drones, such queens last for one and a half to two years of intensive work, due to programmed interbreeding or interbreeding

honey productivity

Families with such queens can be up to 180 percent compared to the average for the apiary. Industrial Queen Bees are sold as proven

Never tested. It is more profitable to buy untested ones, since they are much cheaper, and no more than ten percent of them are rejected. The problem is that they are poorly received by bees. And today there is a simple and reliable method their replanting in the layering, I’ll dwell on it a little. Take a frame with printed brood and place it in an empty hive (housing), there is also a frame with dry land, between them is a cage with a queen,

collateral

A sheet of plywood is placed on the entrance, and the bee is shaken off the three frames where there is open brood (the frames are taken from different hives). We wait until the evening, in the evening we change the empty frame to a honey one and open the feed compartment in the cage. Method

beekeeping

With industrial queens like this.

In the month of June

Layings are made from each bee colony

anti-swarm

On such a queen, and in the fall unites with

basic

Family, the old uterus is removed. I would like to note the main advantages of this technology. This, firstly, checks the quality of the uterus; strong families go into winter, which are good in spring

developed

And they work effectively on early bribes.

As a rule, families

The second year they do not swarm, this is due to the fact that the queens and drones belong to the lines and breeds over which selection for

restlessness

Highly productive daughters F1. To do this, using the method of selecting many parent and maternal families, we select options that best and are guaranteed to transmit their beneficial characteristics to F1 queens, and replicate them. Working with such queens is also very progressive, especially for beekeepers who have more than fifty

bee families

I'll explain. It turns out that you can buy such a queen for your apiary even if you have even a small

core

A parka is much more profitable than buying, say, 50 ordinary commercial queens. In this way we breed high-quality queens,

as a consequence of which

We get families with guaranteed

useful and economic

Using indicators, we also create a certain reserve. The third category includes queens for breeding

These are queens with absolute indicators

Designed for serious research in breeding. The general public of beekeepers do not need them

Firstly

for its price. But from the queens of this category, as a result of painstaking selection work, reproductive queens are determined, but if someone has a desire to develop in the field of selection, this can already be done because reproductive material has recently become available. In conclusion, I would like to express hope and confidence that the days are not far off when Ukrainian specialists in artificial insemination of queen bees will be known not only in the vastness of their native state, but also in leading European countries with developed breeding.

It is known that a young queen needs drones for mating; this occurs exclusively in the air and causes difficulties in breeding work with bees. To control the mating of queens, an isolated flyby is organized (there are no apiaries within a radius of 15 km), but this is very difficult.

For successful insemination in the apiary, there must be enough drones. Of the most strong families paternal families are distinguished in which drones are raised in the early stages. The queen is placed for 2-3 days on a honeycomb (drone cell) in an insulator, which consists of a dividing lattice. Afterwards, the seeded comb is removed from it and placed in the middle of the colony between the combs with open brood. Nowadays, so that drones do not wander around the apiary, and the percentage of selection work is higher, they need to be kept on a frame in an isolator until they reach sexual maturity. To do this, 2-3 days before the drones leave the cells, the honeycombs are placed in an insulator and placed in the hive.

On the day of insemination of the queens, the isolator is brought into a bright room (25-28 ° C), and it is equipped at the artificial insemination laboratory; the drones are released into the window for flying around. As they fly out, they clean out their intestines and gather on the glass; here they are caught, placed in a cage and taken away in order to carry out insemination. The remaining drones are placed in microthermostats for temporary storage and fed with honey.

To obtain sperm from drones, an electric ejaculator is needed; it simplifies and speeds up the collection of semen.

Infertile queens are inseminated on day 5-6. Insemination is carried out on machines made in Czechoslovakia.

Barren queens can be preserved in nucs before and after insemination. The right wing of the queen is cut off to prevent it from flying out for natural mating. Nuclei are a big investment of time and labor. Now the task is to obtain fetal queens without the formation of nuclei.

Before and after insemination, a special queen nursery (A.K. Shushakova) is tested to preserve the queens. Behind the structure is a riser hive, where the honeycombs are arranged in 3 tiers of 6 pieces each. in everyone. Through the wire mesh, the bees can come into contact with the queens, who are kept in special rooms without bees before and after insemination. One such queen nursery can simultaneously contain 108 queens. From two sides, 54 pcs. compartments for the queens are located. Periodically, the queen receiver is reinforced with bees from other families (without a queen). The bees are fed systematically using the top feeder. The resulting infertile queens from the breeding apiary are immediately released into the queen nursery. After 5-6 days they are inseminated and returned to the nursery again. The disadvantage of this content is death large quantity queens both before and after insemination.

Nowadays, in order to preserve the queens, nurse families are used, they are formed in this way. The queen and open brood are removed from the nest of the bee colony, and instead frames with sandy brood from other colonies are added. Frames with beebread and honey are placed on both edges. 9-10 frames are left in the hive, 5-6 of which contain brood. 1 day after the formation of the nursing family, a nursery frame is placed in it where barren queens are located (25-50 pcs.) in shipping wooden cages, which are covered with a film with holes, with bees (7-8 pcs.) and candy. In such a family, the queens stay for 5-6 days. before insemination and 4-5 days. after it, they are then implemented. The contents of queen bees in wooden carrier cages is much better compared to metal ones. The queens in them are in contact with the bees of their family, and the bees of the nurse family only heat the cage.

The most important value artificial insemination of queen bees in that they provide a 100% guarantee of linear affiliation, pure breeding and the success of breeding work.

Artificial insemination of queen bees video

Here is a video where you can inseminate queen bees through selection.

How we achieved success in inseminating queen bees

To develop new lines and breeds of farm animals that have better hereditary potential and produce higher production, livestock breeders widely use breeding work. They cross certain animals with each other according to their choice, obtain offspring from them, check its productivity, cross again, and so in the end they fix the desired qualities in one or another line. In all these cases, the livestock breeder must know well both the father and mother of the resulting offspring. For this purpose, manual mating is used, i.e., creating a meeting between a male and a female in an isolated enclosure under constant human supervision.

In beekeeping, breeding work has so far been almost non-existent in the Union. A small individual beekeeping farm could not create the conditions for its development.

Rice. 1. Uterus brought for artificial insemination surgery

Only now are the first steps towards its organization being taken. Breeding work in beekeeping in the very near future should take on a wide scope and become of great importance.

The beekeeping industry has a number of advantages over others. The queen is capable of laying a mass of eggs, which accelerates the rate of reproduction of material from the breeding queen tens of times; drones inherit properties only from their mother, since, developing from unfertilized eggs, they, as is known, do not have a father. This makes breeding work much easier.

Producers, i.e. queens and drones, as well as worker bees, grow from eggs very quickly: the queen in 17 days, the bee - 21, the drone - 24. Puberty also sets in quickly: in the queen, 7-10 days; in the drone, 10-14 days after leaving the cell.

Despite these features, we also have negative sides. Bees are characterized by one feature that greatly complicates breeding work. This feature lies in the fact that the queen mates with the drone outside the hive and even outside the apiary - in the air. This makes it now impossible to cover the queen with the kind of drone we would like to cover her with, and eliminates the possibility of choosing a male to cover the queen. This feature greatly complicates the ability to create new breeds of bees.

For a long time (more than a hundred years), various researchers have attempted to solve the problem of controlling the mating of queens and drones. Prof. I.M. Kulagin and others proposed releasing selected queens and drones for mating late in the evening, i.e., when the drone period in the apiary usually stops. Others tried to achieve mating in large greenhouses, in rooms, on a leash, in beehives, on ships taken far out to sea, on an isolated island or in deserts, in special mating points, etc.

In all cases, some success was achieved. However, it is clear that with these methods we can only monitor the origin of the drones, but cannot ensure the mating of the queen with one specific drone chosen by us.

Next, attempts were made to introduce either the drone organ directly or drone sperm into the genital tract of the uterus using a pipette or syringe. Such attempts have been made for a long time, including in the former. Russia (P.R. Shumilin, Shirokov, Paleyachuk O.N., Prof. Malyshev, etc.) with partial success.

Rice. 2. Brood of artificially inseminated queen No. 44 during her stay in the nucleus

Pvc. 3. Brood of artificially inseminated queen No. 88

Only in 1926, the American Watson put this matter on a practical footing by constructing a special glass syringe. Inseminating queens with this syringe gave him partial success, although, however, most of the queens, along with fertilized eggs, also carried unfertilized ones.

From 1928 to the present, Watson's method has been tested and applied in the USSR by A. S. Mikhailov, who worked until 1931 at the Tula Beekeeping Station (now the Institute of Beekeeping). He managed to achieve 25% success, but the percentage of queens that carried fertilized eggs after the operation did not exceed 10%.

I.L. Lysenko, who worked with A.S. Mikhailov, in 1931 proposed at the Institute of Beekeeping to inseminate the mother by inserting the inverted penis of the drone into the genital opening of the uterus, expanded with tweezers, and transferring the sperm by further pressing on the abdomen of the drone. fluid (sperm) into the genital tract of the uterus. When tearing off its head, a fully mature drone, as is known, quickly turns its organ inside out. The method itself is not new. However, I. A. Lysenko used a Watson machine for the operation, into which, just as during insemination with a syringe, he tied the uterus with a thread (Fig. 1). Thus it was inseminated in 1931. 13 queens, and of these, 3 queens produced bee brood, or 23% of those operated on. The 1931 experiment resolved the issue in principle and proved the possibility of successful insemination of queen bees by directly introducing the drone's reproductive organ into the genital tract of the queen.

The simplicity of the method and its accessibility raised the question of its verification and further development. The syringe and all the equipment associated with it are now an unnecessary add-on, with the exception of hybridization experiments, when we cannot directly insert the male genital organ, for example, due to its large size, and must use a syringe.

Here it would be appropriate to emphasize the difference between the goals that artificial insemination sets for itself in livestock farming and beekeeping. In animal husbandry, it pursues the goal of more economical use of male sperm to inseminate a larger number of females. We do not have this goal at all.

Rice. 4. Brood of artificially inseminated queen No. 85

On the contrary, during insemination (especially with a syringe), it is sometimes necessary to use the sperm of not one, but two or more drones. Here the goal is to achieve control over mating, i.e., by artificial insemination of queens, what is achieved in livestock breeding by hand mating. This fundamental difference cannot be ignored.

Testing of the method was scheduled for 1932. Unfortunately, I. A. Lysenko refused to go with us and (a team from the Institute of Beekeeping on 20/II 1932 went to the Tsyurupino bee incubation station. No one among us owned the equipment insemination. Nevertheless, this did not bother us and we decided to nevertheless develop the intended topic. D. N. Kozlov and K. A. Muzalevskaya quite soon mastered the technique of so-called “manual” insemination (this is not entirely true, but how, in addition to hands, tweezers take part in the operation) and things went smoothly.

However, with the percentage of luck, things were very bad at first glance. Of the 85 queens inseminated by May 31, only one queen (Fig. 2)—the fourth in order inseminated (No. 4)—engaged and gave exclusively bee offspring.

We knew the reason for this failure and even at the Beekeeping Institute pointed out shortcomings in the technique of inseminating queens. The fact is that before us, both A. Mikhailov and Watson and others took queens 7 days and older for the operation. It was assumed that such uteri were suitable for surgery. But it is clear that success depends on how much we have grasped the state of sexual estrus of the uterus. It is known that the success of artificial insemination of agricultural crops depends entirely on this. animals.

In queens, sperm, as is known, is sucked into a special sperm sac and from there, as needed, is used to fertilize the eggs as they pass through the unpaired oviduct. Nevertheless, we know that under the influence of sexual arousal the queen flies out to mate. Obviously, some kind of maturation of the reproductive system in the uterus, still unknown to us, is taking place. The fact that we do not know the details of this process does not give us the right to ignore it.

Rice. 5. Nuclei, with queens intended for artificial insemination, are equipped with entrance queen catchers of the B. M. Muzalyavsky system

By the end of May, our team accepted D.I. Kozlov’s proposal to inseminate only those queens that stubbornly strive to exit the nucleus through the Hahnemann lattice and thereby prove that they are in a state of stalemate. On May 31, 5 such guarded queens were inseminated. The result exceeded all expectations. Of the 5 queens, 4, or 80%, produced normal bee brood. Moreover, one appeared 3 days after the operations, two after 4 days and one after 14. We then reported this exceptional success to Prof. G. A. Kozhevnikov.

In June, the next batch of 9 queens was inseminated. They gave bee brood 4 or 44%. In the process of work, B. M. Muzalevsky proposed a simple mother catcher. shown in Fig. 5 and 6. With a narrow hole it is inserted into the tap hole of the core, and in the last wide hole a dividing grid is inserted. The bees pass through the grid, and the queen, who has entered under the influence of hunting to mate, not being able to pass through the grid, usually runs along the upper grid, where she is easily noticed by the observer on duty. In July, 15 queens that entered such traps were inseminated, and 7 of them gave birth to bee brood. True, two of these seven queens, after the operations, showed heat again two days later, were re-inseminated, and both bred. Thus, out of the number of operations (15 + 2 = 17), seven ma-toks give 41% success.

In total, out of 29 queens (31 operations), inseminated in heat and tested, 15 fertilized and gave bee brood. Of the number of queens, this is 51.7%, of the number of operations - 48.4%. In total, 33 queens were inseminated in heat, but four of them were killed by bees or died in the first three days after the operation and are therefore considered untested.

We owe our success to the correct use of sexual hunting by queen bees. This can be seen, by the way, from the following figures.

Rice. 6. General view of a group of experimental cores with trenches for mother catchers

T. Kozlov teaches the technique of artificial insemination vol. Tsaregrodtseyu (Vyatka) and Bovina

To the North Caucasian zonal beekeeping station, when inseminating queens using the same method, but without taking into account sexual heat, out of 65, 6 fetuses were obtained (9.5% success). I. A. Lysenko in 1932 at the Institute received 3 fetuses, or 13%, out of 23 inseminated. It is important to note that when queens are inseminated by inserting a drone's sexual organ, almost all queens produce exclusively bee brood. We had 12 such queens out of 16, or 75% (16 is obtained because queen No. 4 was added, inseminated not in heat). We explain it this way: Drone sperm has a certain density. As a result, every drop of it released by a naturally inseminated uterus contains sperm

matozoids. When taking sperm into a syringe, we unwittingly create a mixture of sperm with mucus from the accessory glands. Droplets of such “sperm” often do not contain sperm at all and for this reason some of the eggs of the uterus are laid unfertilized. When introducing an organ, we do not disrupt the composition of sperm.

Some of our queens (No. 4, 71, 96, 89) were transplanted into normal families and they provided brood for incubation. The average daily laying of eggs by queen N» 4 is equal to more than 1,000 eggs over the course of a month, but we do not have the opportunity to discuss this issue in detail in this article.

The tray mother catcher has great methodological significance. It allows you to objectively observe the manifestation of sexual heat, study its dependence on weather, age of queens, time of day and other factors. We collected interesting material on this issue in 1932. There is no doubt that 1933 will allow to finally finalize the work begun and transfer it to socialist beekeeping science affordable way artificial insemination of queen bees, which will make it possible already in the second five-year plan to expand and put selection work in beekeeping at the proper level, to begin serious and promising work on the qualitative improvement of bees.

B. M. Muzalevsky and D. N. Kozlov

Magazine “Beekeeping”, No. 2, OGIZ SELKHOZGIZ, Moscow, 1933.

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Fertilization of the uterus. Mating pope with drones

It is not enough that the bees breed a young queen; she is not suitable for anything until she copulates and becomes fertilized with a drone: only by copulating with a drone does she become completely fertile, i.e. capable of carrying bee and drone eggs, and we call such a fetal queen. If, for some reason, the young queen could not copulate with the drone, then she either remains completely sterile and will not bear any eggs, and we will call such a one a barren one, or she will be able to carry only drone eggs, and we call such a one a drone queen.

So, there are three types of queens: a) fertile, bearing testicles of both sexes, i.e. bees and drones, and so good uterus; b) barn queens, which do not carry any eggs, and c) drone queens, which carry only drone eggs. Both of the latter are useless.

A family with a fertile queen will be good, because such a queen reproduces bees and is constantly maintained in her strength. But the one in which the barn or drone queen will not survive in any case and must die is the same as if it does not have a queen: neither barn nor drone queens reproduce bees, and therefore if the previous bees die out and die, the family must be destroyed .

The queen never copulates with the drone in the hive, but always outside it and high in the air. A young queen with damaged wings and which cannot fly is not suitable for anything. Such a one, having flown out with a drone, will fall to the ground and will not return to the hive, or, feeling its disability, will not dare to fly out, and therefore cannot be fertilized. In both cases, the family must certainly die.

You can easily verify this if you cut off one wing of a young queen that has just emerged from the queen cell and make sure that every time she goes to the drone and falls to the ground, you pick her up and put her back into the hive. This queen will finally stop crawling out, and even if the hive is full of drones, she remains unfertilized and will never carry bee eggs.

The queen copulates in the air not necessarily with a drone from her own hive, but with every one she meets, even from someone else’s hive. Therefore, she can be fertilized, even if there were not a single drone in her hive, as long as there were some in other families. She can even be fertilized by a drone from another apiary, even a distant one, because both the queen and the drones fly more than two kilometers from the apiary.

Due to the fact that the queen cannot be fertilized without a drone, young queens hatched in early spring, when there are no drones in the apiary, must remain infertile. They will constantly fly out to meet the drones, but if they do not meet them within four weeks, they will stop their walks and remain forever sterile, and the family will inevitably die. The queen, which will give birth in the fall, for example in September, is still fertilized, because although at that time the drones have already been expelled, there is still a hive without queens in your own or a neighboring apiary, in which they lingered, and it happens that good families Leave one drone at a time until winter. Therefore, a late-bred queen can find a male and become fertilized, if only good weather permits. The queen copulates with the drone only once in her life and as soon as she is fertilized, she remains fertile forever. During the mating flight or flights, the queen mates with 9-10 drones. A fertilized uterus can have its wings cut off, and this will not harm it in any way, and it remains fertile, because there is no need for it to fly out.

The uterus is fertilized through the copulation of the reproductive parts and the influx of drone semen into the uterus. To receive this seed, she has a special organ, which I will call the seminal receptacle. If you carefully tear apart the last two rings of the abdomen of the uterus, then at the very end you will find a round, chalk-white vesicle the size of a millet grain; this is the spermatheca, into which the drone's seed enters during copulation. In a young, still unfertilized uterus, this vesicle is thin and almost transparent and only when filled with seed becomes round and white. Once you find a dead queen, you can tell by the color of the sperm receptacle whether she is young or old.

So, the drone seed does not fertilize the queen's testicles in the ovary. I have already said above that the queen, even without copulation with the drone, can carry eggs, but only drone eggs, but after fertilization she carries both bee and drone eggs. From this it follows that all the testicles in the uterus are initially of only one kind, i.e. male, or drone, and turn into female, or bee, only when they touch the drone seed located in the seminal receptacle of the uterus. This is accomplished as follows: when the uterus releases an egg, it passes right next to the vesicle with the seed. If at this moment the female moistens it with seed, it will become female, or bee, but if she simply lets it pass, then it will not change and will remain male, or drone. The laying of fertilized and unfertilized eggs by the uterus depends on the size of the comb cells and the pressure of their walls on the abdomen of the uterus. But the unfertilized uterus does not have a seed in the spermatic receptacle, and even if she wanted to, she cannot moisten the testicles with it during laying, and therefore all the testicles coming out of her will be only drone ones. It even happens that a queen fertilized by a drone, which carried bee eggs, turns into a drone, i.e. stops laying bee eggs. This occurs in cases where the uterus is very old and the seed has deteriorated, or when it has lost the ability to control the muscles that move the spermatic receptacle, or when the latter has completely deteriorated from a sting, from strong pressure or from great cold. When a family, numb from the cold, is warmed up and saved, we often see that the queen, who was fertile until then, stops laying bee eggs and will carry only drone eggs.

This theory of the transformation of male testicles into female ones, no matter how strange it may seem, you can give full faith, for the most famous modern naturalists have been positively convinced of it.

When the queen flies out to copulate, beekeepers call it a mating flight with a drone (queen mating flight). Where there are many apiaries, for example in Podolia, during swarming you can often see masses of drones flying in the air in heaps, now rising and now falling down. The common people call these heaps of drones midday because they appear only at noon. These are drones chasing the queen, who has flown out to lose, and beekeepers should pay attention to this, because maybe someone will be able to see the act of copulation, since heaps of drones sometimes fall to the ground and rise again.

The queen makes her first approximate flights on the 3rd day; nuptial flights occur no earlier than the 6th day, usually on the 7th day of life, and then only when there is only one in the hive and the rest of the queens have moved away or been destroyed. The queen usually flies out of the swarm the next day according to the sediment; from the colony that has hatched - three weeks after the departure of the old queen with the first one, when all the bees left after her have bred. The queen flies out only in clear, completely calm weather, and on cold, cloudy and windy days, she remains in the hive. The queens are more likely and more likely to be fertilized in consistently good weather.

The queen flies out between ten and three o'clock, when the drones usually play the most. She repeats this several times a day, sometimes more than ten. Having flown out of the hive and spent several minutes in the air, she returns, then flies out again, and this continues until she meets and copulates with a drone. During this meeting, it remains in the air for a quarter of an hour and at most half an hour, and as soon as it is fertilized after copulation, it no longer flies out.

When the queen returns to the hive after meeting the drone, she happens to notice that the tip of a thin thread is trailing behind her. These are the genitals of the drone, which after copulation came off and remained in the uterus. If you grab such a queen in front of the entrance and carefully pull out these threads from it, you can see through a microscope that these are exactly the same genitals that jump out from the back of the drone when you press it firmly with your fingers. This serves as proof that the queen copulates with the drone in the air. The uterus that has brought such a sign usually does not fly out anymore and becomes fertile. She usually begins laying eggs 2 days after the last mating. Queens that have not started laying eggs on the 10-16th day after leaving the queen cell are found to be Bad quality and are subject to discarding.

The beekeeper will recognize the fertilization of the uterus if he sees eggs in the combs, and eight days later - sealed bee brood.

Not all queens are fertilized at the same time; others copulate on the day of the first flight, another will fly for a week, two, three until it is fertilized. This will depend both on favorable weather and on whether there are drones in the apiary.

Usually the uterus is fertilized within eight days, and the eggs laid by it are already noticeable.

You can observe the flight of the young queen to the drone with convenience if you put a small third one in a box with a capacity of 2.5 liters, with a glass, and place it outside near the window so that through the glass you can see every bee flying out and arriving. The beekeeper does not need such a projectile, because he will see enough of this flight in the apiary with second-rate, third-rate and mature families - if only he wants to pay attention to it.

The flight of the queen to the drone threatens the beekeeper with great disaster, because through this more than one family will become dehydrated, since more than one queen that has flown to the drone will die and will not return to the hive. When the queen takes flight, and there is no young offspring in the hive from which the bees could hatch another queen, the family must be deuterated and die if the beekeeper does not give it another queen. Of course, when flying out for the first time, the queen carefully examines the position of the hive in order to find a way back, but it often happens that in the bustle and crush of the bees during their play at noon, she gets lost and, instead of getting into her hive, ends up in the neighboring one. , especially if this one plays strongly, where other people's bees will grab it and kill it immediately. This happens mostly in apiaries where the hives often stand next to each other, like herrings in a barrel, and even more so when they are similar in appearance. Therefore, at least for the last reason, the hives should be placed as far as possible from one another, because the more they are scattered, the fewer queenless families the beekeepers will have. It doesn’t hurt to give similar hives from which young queens fly out some obvious signs: various tires, straw dressings, pegs driven in near the entrances, etc., so that the queen can orient herself. Color signs are not as useful as shape signs. Since the emerging queen already knows the position of her hive, it should not be rearranged, moved or transferred to another apiary, nor should it be subjected to visible changes until the queen is fertilized and stops flying, otherwise she will get confused and die. The beekeeper should also beware of walking unnecessarily between the hives while the bees are playing, because during the play of the bees, young queens also fly out to meet the drones.

Despite, however, all precautions, more than one queen will die during the flight, because birds can grab her, she herself can fall into the water, but most of all the queens die during the swarm: if the young queen flies out to the drone, and at that moment a swarm will come out and make noise in the apiary, the queen forgets about her hive, interferes with the swarm, where they usually kill her, and her own bees will be left without a queen.

Infertile queens are inseminated at the age of 6-13 days, which corresponds to the timing of their natural mating. Drones become sexually mature after 14 days of age.

A nursery with queens placed in cages is removed from the foster family and brought to the laboratory. In good weather, drones are caught in a cage during their active summer, from 12 to 16 o'clock. During non-flying hours, mature drones are collected directly from the outer combs of the nest. In the laboratory, if the drones have not flown, they are given the opportunity to fly around. This allows

They are able to cleanse themselves of feces and make it easier to invert the endophallus.

Before starting to collect sperm from drones, a syringe is prepared. The syringe sleeve is filled with saline solution (0.9% NaCl) and the tip is screwed in until it is tightly adjacent to the rubber membrane. A small air bubble is drawn into the tip, which separates the liquid from the collected sperm. To induce eversion of the endophallus and ejaculation, the drones are grasped by the head and chest from the ventral side with the fingers of the left hand. The part of the abdomen adjacent to the back is irritated or slightly squeezed with the fingers of the right hand. This operation causes a reduction abdominal muscles and partial and sometimes complete inversion of the genital organ and ejaculation of the drone. If partial eversion of the endophallus (genital organ) has occurred, the drone's abdomen is compressed from front to back until sperm appears. During the complete process of eversion of the endophallus, cream-colored sperm is first released, and then white mucus (Fig. 15).

Rice. 15. Sperm collection from a drone

The drone that has ejaculated is brought to the tip of the syringe, the surface of the seminal fluid touches the tip of the capillary, and as the piston is pulled back, it enters it. Suction of mucus should be avoided, as it quickly coagulates and forms a plug, stopping the passage of sperm. When the next drone is brought in, a drop of sperm from the previous one is released from the tip so that it combines with the new portion, and it is drawn into the syringe. To fill the tip, it is necessary to collect sperm from 6-10 drones. Its selection and filling of the syringe is carried out under a microscope at 8 times magnification. To prevent the sperm in the syringe from drying out, the column is raised and a drop of saline is drawn in.

After collecting sperm into the syringe, they begin to prepare the uterus. Before insemination, she is released into the window, where she flies around and cleanses the intestines of feces. Then the uterus is forced to enter a tube of the same diameter as the uterus holder, the opposite end of which is closed with a finger. Having reached the closed end, the uterus moves back, and at this time a uterine holder is attached to the open end of the tube, into which it moves. When the uterus reaches the narrowed part of the uterine holder, from which the last three abdominal segments should protrude, it is fixed with a stopper with incoming carbon dioxide. Her hind legs should remain inside. Then the uterus is strengthened in the uterine holder block and anesthetized with carbon dioxide.

From the cylinder, carbon dioxide enters a vessel with water through a rubber hose through a reducer. This allows for better control of the gas flow through the air bubbles. From a vessel with water, carbon dioxide penetrates into a rubber hose, at the end of which a plug is attached, fixing the uterus in the uterus holder. In the flow of carbon dioxide, it quickly calms down. It is very important to position it correctly in the block. The central line of the uterine body should be in the same plane as the ventral and stinal hooks. The syringe is inserted exactly along the center line.

As soon as the uterus calms down, they begin to open the sting chamber (Fig. 16, 17, 18). Together with the apparatus, it is placed in the field of view of the microscope at a magnification of 16 times. IN right hand take the auxiliary hook and insert it into the end of the abdomen of the uterus. With a slight movement, the uterine sternite is moved apart and the ventral hook is used to retract it to the left. The auxiliary hook is transferred to left hand and with its help, the sting is retracted to the right with a sting hook, opening the chamber. In the center of the sting is the vestibule of the vagina. The capillary of the tip is directed to the vaginal opening and moved back and forth with light movements to retract the valve that prevents its passage into the unpaired oviduct. Then the capillary is inserted to a depth of 1.5 mm.

If the surrounding tissue begins to move before reaching the required depth, it means that the capillary has entered one of the pockets of the vestibule of the vagina, which are located on both sides. If the semen column does not move, and the air bubble between the sperm and the saline solution contracts, it means that the capillary has not entered the oviduct. In this case, the syringe is pulled back for repeated insertion, the position of the uterus is checked in relation to the syringe and its angle is changed. The sperm is carefully injected, the uterus is released from the uterus holder, placed on the microscope table and, as soon as it begins to move, placed in the cage. If necessary, she is given food and young bees are planted.

After insemination of all queens, the nursery returns them to the breeding family (Fig. 19, 20). With double insemination, repeated manipulations are carried out after 24 hours. High quality queens with triple insemination receive 4 mm3 of sperm at a time. At the end of the last insemination, the uterus, which is in a motionless state, is marked by gluing foil of a certain color to the breast with shellac or BF-6 glue. It takes an average of 10 minutes to inseminate one queen.