Artificial insemination.
For insemination, it is allowed to use stallion sperm, the quality of which fully complies with the requirements of the standard (GOST 24168-80 “Frozen stallion sperm”). Based on this GOST, frozen sperm after thawing and bringing it to a temperature of 20-25 ° C, stored for no more than 15 minutes, according to organoleptic, physical, biological indicators, must comply with the requirements and standards specified in Table 4.
Insemination is carried out in accordance with the Instructions for artificial insemination of mares with stallion sperm stored at a temperature of - 196 ° C (1987).
The use of frozen stallion sperm for artificial insemination of mares is permitted no earlier than 28 days after freezing and in the presence of a document characterizing the quality of sperm according to microbiological and biological indicators.
4. Sperm requirements
Thawed sperm has a low survival rate. When using frozen semen, mares should be bred as close to ovulation as possible. In practice, insemination begins at the III-IV degree of follicle maturity, 12-16 hours before ovulation. Mares are not bred after ovulation.
For insemination, the mare is secured in the pen or by means of a breeding harness and raising the front leg. The external genitalia of the mare are thoroughly washed with a swab soaked generously in clean, warm water. A new swab is used for each mare. The mare's tail from the root to half the rib is bandaged with a clean linen bandage.
Before breeding each mare, the technician thoroughly washes his hands with warm water and soap, wipes them dry with a clean towel and disinfects them with a swab soaked in 96° alcohol.
When thawing sperm frozen in the form of granules, they are placed in one layer on the bottom of a beaker, which is immersed, shaking slightly in a circular motion for 1-2 minutes, in a water bath at 40 ° C until the granules disappear.
When thawing sperm frozen in aluminum bags, they are quickly transferred with liquid nitrogen tweezers for a minute into a water bath at a temperature of 40 ° C. Then the bag is wiped dry with a clean towel, disinfected with an alcohol swab and one end of the bag is opened (can be cut off with sterile scissors).
After thawing, the sperm must be checked under a microscope for sperm motility, which should be at least 2.5 points. The insemination dose (25 ml) must contain at least 300 million motile sperm.
Poorly sealed, burst sperm bags should not be used. During thawing, water can get into them, and then the sperm die.
Mares are inseminated using a sterile elastic rubber catheter, the end of which is inserted by hand into the cervix to a depth of 10-12 cm. The sperm is injected with a syringe, attaching it to the outer end of the catheter.
At the end of insemination, the rubber catheter and syringe are washed with warm water and boiled in distilled water in a special sterilizer.
During the breeding campaign, situations arise when the number of mares assigned to a stallion exceeds the permissible regime for using him in mating. As an additional technique, when carrying out manual mating in stud farms and breeding farms, it is recommended to use freshly obtained undiluted sperm for insemination. One stallion ejaculate can be divided into two or three doses depending on its volume, concentration and sperm motility.
Freshly obtained undiluted sperm is used within 30 minutes from the moment of its receipt.
Mowing mating is the main mating technique used in the herd method of keeping horses. Mares are permanently assigned to a stallion for the entire breeding period, and in some areas for several years.
Moreover, if the stallion was kept separately from mares during the non-breeding season, he is released in the spring to a selected group of mares in a separate base after he has become accustomed to the school and sometimes identifies and covers a mare in heat. The school is driven out to the grazing area assigned to it. In the future, the school stallion covers them as the mares come into heat without human intervention.
During the season, you should not remove mares from the school without reason and add new ones, just as you should not replace a stallion unless absolutely necessary. When forming schools for the next breeding season, it is advisable to preserve the hierarchical connections established in the group, adding only new ones to replace those who have dropped out or replenishing the schools with young mares. This principle of forming schools greatly facilitates the work of herdsmen and helps to increase the foaling rate.
Proper organization of mating allows for high foaling rates. At the same time, the short duration of the breeding period limits the load of queens on breeding stallions.
In cultural herd horse breeding, if there are well-equipped stables for early foaling, it is recommended that outstanding sires be used in hand and cook matings before being released into schools.
Manual mating is technically carried out by bringing the stud stallion by two workers on leashes to a mare in heat, which is held by a third worker. This mating is an effective technique, but quite labor-intensive and is recommended for use along with artificial insemination in stud farms and breeding horse farms, as well as at seasonal points on collective and state farms and when visiting the places where mares are placed on a stud stallion for testing and mating .
The effectiveness of manual mating is highest when follicle maturation is controlled. When monitoring follicle maturation, covering mares should begin at stages III-IV of follicle maturity and repeat after 36 hours until ovulation is established.
If control over follicle maturation is not carried out on the farm, then mating should begin on the second day of heat and repeat after 24, 36 and 48 hours, depending on the survival rates of the sperm of specific stallions until the end of heat and the onset of lights out.
Before mating, the mare must be put on a breeding harness and her tail bandaged. The mare prepared in this way is held by the groom, raising her head so that she cannot hit the stallion. Two other grooms on long leashes lead the stud stallion to the mare. The stallion should not be allowed to mount until he has a fully erect penis. The stallion must remain on the mare until the final ejaculation of sperm, which is indicated by the characteristic twitching of the tail.
If for some reason the landing fails, it must be repeated after 15-20 minutes. After mounting, the stallion and mare need to do a short walk for 10-15 minutes.
A simplified version of manual mating, used in working and commercial horse breeding, is that the stud stallion is brought into the stable stall on leashes, the mares located there are tested and those identified in the heat are covered.
Cooking mating is carried out in a fenced area without human intervention. It differs from manual in that it is less labor intensive, and from school in the instability of the composition of the queens and the frequency of presence of a stallion among them.
For the wark mating, groups of mares assigned to the stallion are driven into a spacious warok (pen, base), where the stallion is released. After mating, the stallion is removed from the breeding, and the queens are released to pasture. This mating is used in cultural herd horse breeding, when the mares are poorly horned and the stallion for some reason cannot be released into the school.
Currently, this technique is widely used in working horse breeding due to the small number of livestock and limited service personnel.
Ideally, a stud stallion used for on-farm work is released daily into the breeding stall overnight, where he finds a mare in heat and covers her. The constant presence of a stallion with mares at night does not violate the established hierarchical connections in a group of horses, which minimizes the possibility of injury. Using a stud stallion at work makes him calmer and helps reduce the number of matings per mare, which prevents his sexual exhaustion. It is possible to keep the stallion in the stall 24 hours a day without using it for work.
There are also additional options for cooking mating:
releasing a mare, presumably in heat, and a stud stallion and removing them after mating;
release of a stud stallion into the breeding facility, where among others there is a mare presumably in heat, and removal of the stallion after mating.
When using boil mating techniques, the stud stallion should not be released into a group of horses where there are recently castrated geldings and uncastrated stallions two years old and older.
ARTIFICIAL INSEMINATION
Artificial insemination is the process of introducing a certain dose of sperm into the female genital tract using special instruments.
Advantages of artificial insemination
A) the volume and quantity of sperm required to fertilize one female is significantly reduced.
B) sperm from valuable producers can be stored for a long time and used for several decades.
C) it is possible to coordinate breeding in all regions
D) through the selection and selection of parental pairs, purposefully obtain the desired changes and characteristics.
The effectiveness of artificial insemination is manifested only in combination with adequate feeding, proper maintenance and operation.
Physiological basis of artificial insemination
The technique of artificial insemination of female animals of different species is based on the structural features of the genital organs. In addition, the speed of sperm movement, depending on the condition of the female, is of particular importance. The closer the moment of ovulation, the higher the nervous and hormonal tone of the genital tract, the faster the sperm move.
During ovulation, follicular fluid enters first the oviduct, then into the uterus and brings sperm into an active state. In the genital organs of cows, sheep and rabbits, sperm remain viable for an average of 36-48 hours, in pigs - 24-48 hours, in mares up to 90 hours, in dogs up to 6-7 days. Bird sperm, unlike animal sperm, lasts up to 35 days or more.
Time of insemination of females
After ovulation, the eggs in the oviducts retain the ability and fertilizing ability to fertilize for only 2-6 hours, in dogs 2-3 days. In cows and heifers, follicles (unlike others) ovulate 10-15 hours after the end of the heat, so they are inseminated in the middle and at the end of the heat. In mares, sheep, goats, pigs and dogs, follicles ovulate spontaneously during the second half of heat. A characteristic feature of rabbits is provoked ovulation: mating is necessary to rupture the follicles. In poultry, eggs are formed spontaneously throughout the entire laying period (season).
Methods of artificial insemination of females. During artificial insemination, the introduction of sperm basically imitates the uterine type in all females. When inseminating cows (heifers), sheep, goats and rabbits, sperm is injected into the cervix (cervical method), where the conditions for sperm survival are most favorable. When inseminating mares, sows and dogs, sperm is injected directly into the uterus (body of the uterus).
Veterinary and sanitary rules for artificial insemination
In order to prevent the risk of spreading diseases, the following rules must be strictly followed:
A) technicians for artificial insemination of animals in clean gowns, caps and scarves, and in farms unaffected by infectious diseases in aprons and rubber boots
B) instruments for artificial insemination, instruments, utensils and equipment must be sterile
C) syringes - catheters are sterilized in disassembled form: boil for 15-20 minutes with the sterilizer closed.
D) the vaginal speculum can be disinfected with 96% ethanol followed by washing with a sterile solution of 1% NaCl.
D) before artificial insemination, females (cows, heifers, mares, sheep, goats, sows) are fixed in special pens (or stalls). The external genitalia are washed with clean water from an Esmarch mug, irrigated with a solution of furacillin (1:5000), furazolidone (1:1000), wiped dry, the labia of the females are opened and sterile instruments are inserted. The vaginal speculum must be pre-moistened with a warm, sterile 1% sodium chloride solution.
Artificial insemination of cows
Cows and heifers are inseminated only if they are in heat twice: the first time immediately after detection of heat and the second time after 10-12 hours (if the heat continues). Single insemination is also allowed, but only with rectal control of the maturation of follicles in the ovaries or detection of heat by a test bull. Sometimes they practice double insemination during one hunt with an interval of 10-15 minutes.
Currently, 4 methods of artificial insemination of cows and heifers are used: vaginal - sperm is injected into the vagina or onto the cervix, visocervical - using a vaginal speculum, rectocervical and manocervical.
When inseminating cows and heifers with freshly obtained sperm, they work in a laboratory or arena at a temperature of 18-20ºС. The sperm is drawn into a warm syringe-catheter or an insemination polystyrene pipette. The catheter is lowered into a bottle with sperm and it is slowly drawn into the syringe. Then, turning the syringe with the catheter up, moving the piston downwards collects all the sperm from the catheter channel into the syringe barrel. Without changing the position of the syringe, the piston is carefully moved upward, displacing air bubbles from the cylinder until a drop of sperm appears at the end of the catheter, which is applied to a glass slide to evaluate sperm activity a second time. If the quality of the sperm has not decreased, insemination begins.
If cows are inseminated with sperm cooled to 0-4 ºС, then it is drawn into a pre-heated syringe catheter, for which the latter is washed several times with a sterile 1% NaCl solution or 1% sodium bicarbonate solution t 42-45 ºС. In this case, t of the syringe = 35-40ºС. And the sperm collected in it is 30-35ºС
Granulated sperm, located in ampoules or vials, after thawing, is immediately placed in the nests of a tripod made of foam rubber. Sperm granules with a volume of 0.5 ml are thawed in a sterile foakon. Using a sterile carnzang, two granules (dose) are taken out of the Dewar flask and placed in warm water in a thermos at 40-42ºC, rotating the bottle until the two granules are completely thawed. With a granule volume of 0.5 ml, the thawing time for two granules is 1.5-2 minutes. You cannot put several doses into one, i.e. 4, 6, or more granules, because the defrosting mode is disrupted. The fertilizing ability of sperm also decreases.
Sperm frozen in lined granules is thawed in a thermostat at a water temperature of 38-40ºC for 8-10 seconds.
After the sperm has thawed, wipe the lining dry with a sterile cloth and check for leaks by applying light pressure. After making sure that the tightness of the lined granule is not broken, the latter is placed in the body of a disposable instrument for the manocervical method of administration, then the pusher pushes it to the front edge of the body until it stops. Through the outlet of the instrument body, a sterile needle is used to pierce the shell of the granule and the sperm is injected into the genital tract of the cow.
If granules of concentrated sperm with a volume of 0.1-0.2 ml are used, then it is thawed in a sterile 2.9% sodium citrate solution in ampoules with a volume of 0.8-1 ml, while placing it in warm water (40-42ºС) for 5 -8 sec. Only one granule is thawed in one ampoule. The ampoule with thawed sperm is immediately removed from the vessel with water and placed on the table in the tripod sockets at room temperature 18-20ºC or placed in a thermos.
When sperm is frozen in straws or paillettes in a dose of 0.2-0.25 ml, it is thawed in a thermos or thermostat at water temperature = 38ºC for 9-10 seconds. The straw is removed and cleaned of water, carefully wrapped in a paper napkin and blotted.
Visocervical insemination
For this method, use 4 numbered glass jars (No. 1,2,3,4) per 100 ml with ground-in lids. A sterile 15% sodium chloride solution (2.8% sodium citrate solution is possible) is poured into jars No. 1, 3, 4, and 70% ethyl alcohol is poured into jar No. 2; the solution in jars No. 3 and 4 must be warm (38-40ºC) so that the syringe catheter is heated before filling it with sperm. The syringe is washed with a solution 3-4 times from jar No. 1 and after disinfection with 70% ethyl alcohol from jar No. 2, washed with a solution from jars No. 3 and 4 (removing residual alcohol)
After collecting sperm, the syringe is held vertically, with the catheter facing upward. A prepared vaginal speculum, moistened with a warm (38-40ºC) sterile 1% sodium chloride solution, is inserted into the female’s vagina (the external genitalia are first treated with warm water, furazolidone solution, dried with a napkin or cotton wool. The vaginal speculum is inserted, turned with the handles down, carefully opening the jaws and, having found the cervix, a syringe-catheter or an insemination pipette is inserted into the canal to a depth of 4-6 cm.
Insemination using the rectocervical method
Before insemination, instruments are prepared as follows:
The corner of the bag with disposable sterile pipettes is wiped with a swab moistened with 96% alcohol and broken through with a pipette;
Having extended the pipette to 1/3 of its length, it is connected to the syringe and removed from the bag.
If sperm frozen in paillettes is used, then a special tool is used consisting of a cover, a metal cylinder with a piston, 45 cm long, and a fixation spring. The bag with protective covers is also treated with a swab moistened with ethyl alcohol, the corner of the bag is cut off, a protective cover is put on a previously prepared syringe with an inserted sequin and secured with a spring (before this, the tip of the sequin directed outward is cut off).
The instruments prepared for insemination are placed on a special stand. A drop of sperm is squeezed onto a glass slide and must be checked under a microscope for activity.
Before inseminating a cow (heifer), put a disposable glove on your hand, moisten it with warm water (preferably soapy) and open the external genitalia of the cow, and with the other hand insert a pipette or insemination instrument with a sequin at an angle of 25-35º into the vagina up to the cervix. Then a hand in a plastic glove is inserted into the rectum and the cervix is found, fixing it between the index and middle fingers. Using your thumb or little finger, guide the instrument into the cervical canal to a depth of 6-8 cm, slowly press on the syringe plunger or sperm ejector from the separator.
Insemination using the manocervical method
Initially, as is customary with any method, a thorough toileting of the cow’s external genitalia is carried out. Next, take the ampoules with sperm out of the thermos or thaw them if it was stored in liquid nitrogen, wipe it with an alcohol swab, cut off the cap of the ampoule with sterile scissors, and squeeze a drop of sperm onto a glass slide to evaluate the activity. Then the ampoule is connected to the catheter. Without removing it from the plastic bag.
Put a glove on your hand, moisten it with a warm 1% sodium chloride solution, insert your hand into the cow’s vagina, feel the cervix and determine its condition. Then the hand is retracted to the vestibule of the vagina, with the other hand the prepared ampoule with the catheter is fed into the hand located in the vagina, the catheter is inserted into the cervical canal to a depth of 4-6 cm. The sperm is squeezed out with the thumb and forefinger from the bottom of the ampoule towards the catheter. Without opening the ampoules, carefully remove the catheter from the canal and perform a light massage of the cervix
ARTIFICIAL INSEMINATION OF SHEEP AND GOATS
Sheep and goats are inseminated visocervically with diluted or undiluted sperm at artificial insemination points, both freshly obtained (stored at a temperature of 2-4ºC for 2 days with an activity of at least 8 points), and frozen at a temperature of -196ºC (after thawing, sperm activity must be at least 4 points).
The goat or sheep is brought into the pen and the external genitalia are treated with a napkin moistened with a 1:500 solution of furacillin. Before insemination, the female's vagina is examined using a sterile, warm vaginal speculum to check for signs of disease - rash, blood, pus. The branches at the cervix are slightly opened. Then the syringe catheter is inserted into the cervical canal to a depth of 1-2 cm; the plunger is pressed with the thumb so that sperm does not flow into the vagina; before pressing the piston, the speculum is slightly pulled back. Before inseminating the next sheep (goat), the syringe catheter is wiped with a swab soaked in 96% alcohol so that alcohol does not get into the cannula.
After each sheep, the vaginal speculum is washed with a warm 2-3% sodium bicarbonate solution, wiped dry and disinfected (flamed). When all the sperm is used up, before filling the microsyringe with sperm from another ram (goat), the syringe is washed with a 1% solution of sodium chloride (jar No. 1), then with 70% alcohol (jar No. 2) and the alcohol is washed off (from jars No. 3 and No. 4).
After use, catheter syringes are washed with warm water and sterilized by boiling (you can leave 70% alcohol in the syringe until the next insemination).
Sheep (goats) are inseminated twice in one hunt. Sexual heat is detected by a sampler ram 2 times a day - in the morning and in the evening. Sheep whose heat lasts more than a day must be inseminated twice: the first time immediately after sampling and the second time a day later (after each insemination, the sheep are marked with easily washable paint on the croup). Sheep and goats in which the cervix is difficult to detect can be inseminated vaginally (paracervically). A syringe catheter with a shortened end is inserted deeply into the female’s vagina and undiluted sperm is squeezed out in a dose of 0.1 ml, diluted in a dose of 0.2 ml.
A new flock is formed from the inseminated sheep, and from the 12th day after insemination with a sample ram, sheep that have come into heat again are selected.
ARTIFICIAL INSEMINATION OF PIGS
Unlike artificial insemination of cattle, where imported thawed semen is mainly used, in pig breeding producers are often kept on farms. Therefore, to organize insemination, sperm is first obtained from the boar. As a rule, the taking regimen is once every 5-7 days.
If the sperm is going to be used for a long time, it must be frozen. Therefore, after taking it, it is filtered from the secretion of the Cooper glands. The filtrate is kept in the dark at a temperature of 18-20ºC for 60 minutes. For freezing, ejaculate with sperm activity of at least 7 points and a concentration of at least 150 million/ml is used.
After holding for an hour at a temperature of 18-20 ºС, the sperm is heated to 30 ºС, diluted in a 1:1 ratio with a medium heated to the same temperature. Then the sperm is kept at an additional temperature of 18-20 ºС for 4 hours, after which it is placed in the refrigerator (10 ºС) and after 30 minutes. The flask with sperm is transferred for 10-15 minutes into melting ice and water (0 ºС).
If the sperm is frozen in granules on a block of dry ice, the diameter of the wells is 1 cm, the volume of the granules is 0.5 ml, after 5 minutes the granules are transferred to liquid nitrogen for storage.
The granules are defrosted in a special device with water heated to 42-43 ºС. The device is used to separate the liquid phase of sperm from the solid phase at the moment of its formation.
Sperm activity is about 4-5 points. Sows are inseminated twice in one heat with a dose of 100 ml containing 3-5 billion motile sperm.
When inseminating pigs, boar sperm is injected directly into the uterus. During use, sperm should be at a temperature of 30-35 ºС, because the cold sperm causes the uterus to contract and is expelled into and out of the pig's vagina. Nowadays, POS-5 polyethylene devices are widely used.
Pigs are inseminated at points in individual pens-cages or directly in the pens where they are kept. The external genitalia of the female is treated with a solution of furacillin 1:5000. the catheter is inserted into the pig’s vagina without a vaginal speculum until it stops at the cervix to a depth of 25-30 cm, then directly into the cervix (which periodically contracts), so the sperm must be poured into the uterus within 3-4 minutes. The dose of sperm is 1 ml per 1 kg of live weight of the animal, but not more than 150 ml (3-5 billion active sperm). The fractional method of artificial insemination of pigs involves first introducing thinly diluted (1:1) or undiluted sperm into the uterus in a volume of 40-50 ml (2-3 billion motile sperm), and then a diluent (glucose-salt) - 70-100 ml. For this method, a special UZK-5 probe is used, consisting of a metal catheter with a head at the end, which is connected to vials for sperm and diluent. The bottles are connected by rubber tubes to Richardson balls. Bottles of 100-250 ml are placed in heated wooden boxes. After inserting the probe into the uterus, I open the bottle with sperm and, using Richardson balloons, force the required dose of sperm into the cervical canal with air; The bottle with sperm is closed, the bottle with diluent is opened and a certain dose is administered.
Sexual heat in sows is detected in the morning and evening using a test boar in pens or courtyards specially designated for this purpose (immobility reflex). Sows in heat are inseminated immediately and then again after 20-24 hours.
ARTIFICIAL INSEMINATION OF MARES
Preparation for insemination of a mare is always accompanied by additional measures to prevent injury. Therefore, to begin with, the mare is brought into the stall or held by the reins; in addition, during the injection of sperm, the mare’s forelimb is raised so that she cannot hit her hindlimb. First, the root of the tail is bandaged. The groom takes the mare's tail to the side and washes the external genitalia from Esmarch's mug with a solution of furacillin 1:5000. Mares are inseminated in two ways: vizocervical and manocervical. The sperm is injected directly into the uterus (to a depth of 10-20 cm) using a rubber, ebonite or polystyrene catheter.
Artificial insemination of mares can be carried out in two ways.
For the manocervical method, rubber and polystyrene catheters are used. The veterinarian inserts a prepared hand into the mare’s vagina, the end of the catheter, and moves it through the cervix into the body of the uterus; the assistant lifts the syringe or ampoule and injects the sperm.
For the visocervical method of insemination, both ebonite and polystyrene catheters designed by the All-Russian Research Institute of Horse Breeding are used. The catheter is a tube 50 cm long, 0.6 cm thick, one end of which is expanded in the form of a head. The catheter is connected to a syringe or ampoule using a rubber coupling with a clamp. The catheter is used simultaneously with a vaginal speculum, which is sterilized and moistened with a 0.9% NaCl solution.
Mares are inseminated freshly obtained or stored at 2-4ºC for 3 days or thawed. When thawing sperm frozen in aluminum bags, they are quickly transferred with liquid nitrogen tweezers to a water bath at 40ºC for 1 minute. The dose of freshly obtained sperm is 25-30 ml. The dose should contain at least 150-300 million sperm with PPD. The activity of freshly obtained sperm is not lower than 6 points, thawed - 2 points.
Sperm is injected only when heated to 30-35ºС. After 8-9 days, inseminated mares are checked by a test stallion for the presence of heat. 35-40 days after insemination, mares are checked for pregnancy using the rectal method.
ARTIFICIAL INSEMINATION OF BIRDS
Insemination of birds is organized in the afternoon (after egg laying): chickens - once every 5 days, geese - once a day and turkeys at the beginning of the season 2-3 times with an interval of 1-2 days, then 10-12 days.
To ensure the efficiency of the insemination process, the work of the teams is divided into 2 groups. The first group (2 people) receives sperm from roosters, the second (2-3 people) inseminates hens. In 1 hour you can inseminate 120-150 chickens. The assistant holds the chicken with his left hand, and with his right hand presses lightly on its stomach in the area between the pubic bones and the chest, where the oviduct is located. At the same time, the bird's cloaca is slightly extended. With his left hand, the veterinarian slightly stretches the cloaca with a cotton swab moistened with a 1:5000 furacillin solution until the opening of the oviduct, located slightly to the left, appears. With his right hand, he inserts a catheter into the oviduct to a depth of 4-5 cm and squeezes out a dose of sperm.
The dose is 0.02, 0.03 and 0.05 ml and the sperm count is at least 10 -15 million, with an activity of 7 points.
ARTIFICIAL INSEMINATION OF RABBIT
The main feature of artificial insemination of rabbits is that ovulation occurs only as a result of irritation of the vaginal receptors. Therefore, the female rabbit is first mated with a vasectomized male. Then they are fixed on a special machine with their back down. A cotton swab moistened with a 1:5000 solution of furacillin is used to treat the external genitalia of the female. Freshly diluted sperm (0.3 ml) or stored for 5-6 hours at 0ºC (0.4 ml) with a score of at least 6 points and a content of 5-10 million is collected into a syringe catheter (you can use a shortened one for inseminating sheep) sperm per dose. You can use sperm, thawed in a thermostat at 38ºC, in a dose of 1 ml with an activity of 3 points, containing at least 4.5 million motile sperm.
During insemination, the fingers of the left hand slightly spread the female’s genital slit, and with the right hand a syringe-catheter is inserted, pointing first down and then into the vagina, behind the pubic fusion it is turned parallel to the axis of the female’s spine. When inserting the syringe, sudden movements should not be allowed. Sometimes the catheter syringe is difficult to pass into the vagina as a result of spasm. In this case, you need to pause, after which the catheter is inserted at 12-14 cm and sperm is injected at the cervix (double type of uterus).
Artificial insemination of horses is carried out by state stations for breeding and artificial insemination and state factory stables, which are staffed with highly valuable breeding stallions - elite producers. Insemination of mares on collective and state farms is carried out at artificial insemination points, where the sperm of the producer assigned to the farm is transported from the state breeding station or from the state stable. State and collective farms can organize insemination points located within a radius of up to 10 km. State stables place stallions at such points. If farms are located more than 10 km from the point, sub-points are organized in them, where sperm is brought from the main point. To carry out work at the point, the board of the collective farm (directorate of the state farm) allocates an artificial insemination technician, a groom and a cleaner. Insemination of mares at the sub-point is assigned to the technician of the main point. Only technicians who have received special training in courses have the right to carry out artificial insemination of horses. Artificial insemination stations for horses are located in standard or adapted premises. They must have an arena, a laboratory, a washing room, a storeroom, a stable, a room for storing harness and fodder, and a walking area for stallions.
The area of the arena for collecting sperm and inseminating mares should be approximately 50 m2, its height should be 3.75 m, the light area should be 1:10. The floor of the arena is covered with soft asphalt and a drain is made for the liquid. It is recommended to cover the walls with light-colored oil paint. To secure mares during insemination, a wooden or metal machine is installed in the arena. The laboratory for research, dilution and storage of sperm is located in a well-lit, heated room. Its walls are painted with light oil paint. The air temperature in the laboratory during the cold season must be maintained within 18-25°. The washing room should be located between the laboratory and the arena. In it, dishes are washed and sterilized, and the vagina is prepared to receive sperm. The washing room must have hot water and a drying cabinet. At subpoints where mares are only inseminated, a heated room is equipped for storing sperm, tools and preparing utensils and equipment, as well as a separate room (manege) for inseminating mares.
Receiving sperm from stallions
Preparation of stallions. A month before the start of the breeding season, stallions are examined for the condition of their reproductive organs and the quality of sperm is checked three times at one-day intervals. The indicators of sperm obtained on the third day of the study are considered decisive. Satisfactory indicators of the quality of a stallion's sperm are the initial activity of sperm movement not lower than 5 points, their concentration in 1 ml of undiluted sperm is 250 million and above during testing after a long break in mating and 150-250 million sperm with regular sperm collection. Survivability of sperm at a temperature of 2-4°C in a glucose-yolk medium is 6-8 days. Good maintenance, feeding and proper use of breeding stallions increases the quality of their sperm and its fertilizing ability. Stallions at stations and artificial insemination points are recommended to be given one cage per day (six cages per week). Repeated plantings are allowed in exceptional cases. Young and old stallions have their sexual load reduced to four to five cages. The intervals between cages must be at least 24 hours. The stud stallion should not be used as a sample.
Sperm collection device. To obtain sperm from a stallion, an artificial vagina is used, which consists of a frame, a chamber, fixing rings and a sperm receptacle. The frame of the artificial vagina is an aluminum cylinder with a diameter of 13 cm. At one end, the cylinder is narrowed in the form of a neck, ending in a short cylinder of smaller diameter. The length of the small cylinder is 7.5 cm, the diameter is 8.7 cm, the length of the neck is 4.5 cm, and the length of the entire artificial vagina is 54 cm. A pipe with a screw-on nut is inserted into the middle part of the large cylinder. There is a valve in the nut to relieve excess pressure generated during the stallion's eddy. An aluminum handle is also soldered to the large cylinder. A rubber bladder is placed on the metal frame and secured to it with three rubber rings. A rubber sperm receptacle, which is a wide rubber cup, is placed on top of the rubber chamber onto the small cylinder.
The artificial vagina is prepared as follows: the rubber chamber is thoroughly washed from talc, then, after drying, it is put on the frame so that the smooth surface of the chamber faces the inside of the vagina, and the rough surface faces the frame. You should not tighten the chamber too much, given that when you subsequently pour water into the artificial vagina, the walls of the chamber fall off. The put on rubber bladder is pressed from the outside to the frame with rubber rings. Then they are disinfected by wiping with cotton swabs soaked in 96° alcohol.
With such a swab, using a long forceps, they wipe the surface of the chamber inside the artificial vagina, the inner surface of the previously washed and dried sperm receptacle, and that part of the surface of the artificial vagina on which the sperm receptacle will be placed. After the alcohol has evaporated, the sperm receptacle is placed on the neck of the artificial vagina. Through the pipe, 1.5-2.5 liters of water heated to 45-60° (depending on the ambient temperature) are poured into the space between the frame and the rubber chamber, with the expectation that the temperature in the vagina is 40-42°. It is imperative to check the temperature immediately before insertion, for which a chemical thermometer is inserted into the open end of the slightly inclined artificial vagina so that it fits snugly against the collapsed walls of the rubber chamber. Using a previously disinfected forceps, the inner tube of the vagina is well lubricated along its entire length with sterile fusible petroleum jelly.
Receiving sperm. To obtain sperm, a mare in heat or a phlegmatic disposition is used. A breeding harness is put on the mare (so that she cannot hit the stallion), and the tail from rib to half is bandaged with a linen bandage so that the hair of the tail does not interfere during mounting. When mounting the stallion, the artificial vagina is held with both hands, pressing it tightly against the mare's croup on the right side and tilting the wide end of the vagina 30-36°. With your right hand you need to firmly rest against the sperm receptacle so that when the stallion’s penis pushes, the artificial vagina is motionless. At the moment of mounting, if the walls of the rubber tube of the vagina are very tense, it is necessary to turn the nut of the pipe. This opens the valve located in it, and excess air leaves the artificial vagina. At the end of ejaculation, the sperm receptacle is gradually lowered down so that the sperm does not leak out of the vagina. After receiving the sperm, the sperm receptacle is removed, covered with a sterile napkin and transferred to the laboratory. Here the sperm is immediately filtered through sterile gauze into a graduated beaker pre-warmed to 25-30° and covered with a Petri dish. The gauze with the thick viscous secretion of the accessory sex glands remaining on it is thrown away. Water is poured out of the artificial vagina, the rubber chamber is thoroughly washed from Vaseline and sperm residues with a warm soda solution, rinsed with warm water, wiped with a clean towel and dried.
Preparation for insemination of a mare is always accompanied by additional measures to prevent injury. Therefore, to begin with, the mare is brought into the stall or held by the reins; in addition, during the injection of sperm, the mare’s forelimb is raised so that she cannot hit her hindlimb. First, the root of the tail is bandaged. The groom takes the mare's tail to the side and washes the external genitalia from Esmarch's mug with a solution of furacillin 1:5000. Mares are inseminated in two ways: vizocervical and manocervical. The sperm is injected directly into the uterus (to a depth of 10-20 cm) using a rubber, ebonite or polystyrene catheter.
Artificial insemination of mares can be carried out in two ways.
For the manocervical method, rubber and polystyrene catheters are used. The veterinarian inserts a prepared hand into the mare’s vagina, the end of the catheter, and moves it through the cervix into the body of the uterus; the assistant lifts the syringe or ampoule and injects the sperm.
For the visocervical method of insemination, both ebonite and polystyrene catheters designed by the All-Russian Research Institute of Horse Breeding are used. The catheter is a tube 50 cm long, 0.6 cm thick, one end of which is expanded in the form of a head. The catheter is connected to a syringe or ampoule using a rubber coupling with a clamp. The catheter is used simultaneously with a vaginal speculum, which is sterilized and moistened with a 0.9% NaCl solution.
Mares are inseminated freshly obtained or stored at 2-4ºC for 3 days or thawed. When thawing sperm frozen in aluminum bags, they are quickly transferred with liquid nitrogen tweezers to a water bath at 40ºC for 1 minute. The dose of freshly obtained sperm is 25-30 ml. The dose should contain at least 150-300 million sperm with PPD. The activity of freshly obtained sperm is not lower than 6 points, thawed - 2 points.
Sperm is injected only when heated to 30-35ºС. After 8-9 days, inseminated mares are checked by a test stallion for the presence of heat. 35-40 days after insemination, mares are checked for pregnancy using the rectal method.
In order to prevent the risk of spreading diseases, the following rules must be strictly followed:
A) technicians for artificial insemination of animals in clean gowns, caps and scarves, and in farms unaffected by infectious diseases in aprons and rubber boots
B) instruments for artificial insemination, instruments, utensils and equipment must be sterile
C) syringes - catheters are sterilized in disassembled form: boil for 15-20 minutes with the sterilizer closed.
D) the vaginal speculum can be disinfected with 96% ethanol followed by washing with a sterile solution of 1% NaCl.
D) before artificial insemination, females (cows, heifers, mares, sheep, goats, sows) are fixed in special pens (or stalls). The external genitalia are washed with clean water from an Esmarch mug, irrigated with a solution of furacillin (1:5000), furazolidone (1:1000), wiped dry, the labia of the females are opened and sterile instruments are inserted. The vaginal speculum must be pre-moistened with a warm, sterile 1% sodium chloride solution.
Breeding zootechnical selection of mares to stallions during artificial insemination is of great production importance.
Skillful and well-founded breeding selection raises the production efficiency of the artificial insemination method even higher, ensuring the production of offspring of the best shapes and qualities in large quantities and in the shortest possible time. And, conversely, incorrect breeding selection or its absence greatly reduces the importance of the artificial insemination method as the main and indispensable zootechnical measure for the massive and rapid improvement of the qualities of a horse.
On this very serious issue one hears such statements from individual livestock specialists and horse breeders.
Artificial insemination in breeding, and even more so in elite (purebred and thoroughbred) horse breeding is not applicable, since in these cases the main role is played by strict individual zootechnical selection of mares to stallions; there are no and cannot be conditions for selecting a significant number of mares for one stallion.
Breeding selection in purebred and thoroughbred horse breeding is based on a strictly individual zootechnical approach to each animal, and therefore artificial insemination with its principle of mass use of stallions in mating will not find application here.
3. Artificial insemination can be widely used in production only for the accelerated improvement of horses of local breeds in the collective farm sector of horse breeding, where selection is not as strict and mandatory and does not have such great and decisive importance as in breeding.
We cannot agree with such statements. Artificial insemination in combination with breeding selection is necessary not only for mass and breeding, but also for elite purebred and purebred horse breeding.
An example is selection work with horses of the Budennovsky breed.
Among the horses of this breed there are a lot of elite mares and stallions, but especially outstanding producers of super-elite importance such as Sleding (Simpatiaga-Julieta), Symbol (Simpatiaga-Volna), Sagar (Simpatiaga-Garantiya), Saksagan (Simpatiaga-Garantiya), Ka- hum (Cocas-Agave), Imam (Inferno-Madame Ango), not so much.
These outstanding stallions, the founders of the breed, during natural mating, inseminate annually, depending on their sexual potency, 15-28 mares, and meanwhile, for each of them, taking into account genealogical, production, interior and exterior data, a significantly larger number of mares can be selected and, using the method artificial insemination, to have several times more valuable offspring from them.
The outstanding Don stallions Dorogoy, Bolivar and others can be matched with 100 mares or more annually. Obtaining an increased number of offspring from such outstanding sires would be a most valuable contribution to breeding work with horses of the Don and Budennovsky breeds.
Thus, there is every reason to use the method of artificial insemination in combination with zootechnical selection in pedigree and elite purebred horse breeding. The same can rightfully be said in relation to thoroughbred riding and purebred trotting and working-harness horse breeding.
Is it not possible to select 60-100 purebred mares annually for such outstanding quality purebred riding stallions as Brimstone, Tagore, Cyclonic, Granit II, Marseille, Zagar, etc., artificially inseminate the latter with the seed of these stallions and obtain the maximum of the most valuable offspring?
Of course, it is not only possible, but also must. Here is some factual data on this matter.
In 1939, at the stud farm named after the 1st Cavalry Army, the seed of the best purebred riding stallion Bode-ler was artificially inseminated according to strict individual breeding selection of 14 purebred riding mares, 136 elite and breeding mares, and 110 replacement mares according to group breeding selection. Of the total number of 260 mares inseminated with the semen of the stallion Beaudelaire, 95% were foals. The offspring born in 1940 from Beaudelaire (Fig. 17) turned out to be very good overall.
In 1949, the most valuable Don stallions were used by artificial insemination with only a smaller load of mares: Dorogoy (champion BGXB-1939), Bolivar (reserve champion BGXB-1939), Boston (1st degree diploma of the All-Russian Agricultural Exhibition-1939) , Dvinsk, Barvinok, Sagar, Cahul, etc.
Group selection, when mares are assigned to one or two stallions based on their exterior, genealogical and other qualities, is applicable only to replacement mares and in the mass sector of horse breeding.
Even with a small number of highly valuable breeding and elite mares in the stud, it is quite possible with
Rice. 17. Purebred riding stallion Bodelaire, one of the founders of the Budennovsk horse breed. In 1939, 260 mares were inseminated with his seed at the stud farm named after the 1st Cavalry Army, of which 95.3% were fertilized.
By using the artificial insemination method, increase their workload (50-90 or more) for breeding selection for outstanding stallions. We mean the transfer of mares and stallions from one stud to another for the breeding season, where better and wider breeding selection is possible, as well as the grouping of mares in studs by line and by stallion.
With the wider use of the method of artificial insemination in breeding and elite business, of course, a significant regrouping of mares into herds, sections, stallions and even, perhaps, into studs will be required.
Only in this way is it possible to make the most rational use of the best stud stallions and obtain from them the maximum of valuable offspring.
Under the conditions of socialist agriculture there is every opportunity for such a rational restructuring of the breeding business. The successfully applied method of storing and transporting stallion semen over considerable distances opens up wide opportunities for maximum use of the best producers. It goes without saying that transferring mares for insemination from one site (plant) to another is permissible only if there are no veterinary and sanitary contraindications to this and if it is sufficiently justified.
Some horse breeders are inclined to think that breeding selection during artificial insemination can encounter great difficulties.
These difficulties, in their opinion, may arise in connection with the breeding selection of a large number of mares to individual stallions, as well as in connection with the replacement of some stallions by others in order to prevent closely related breeding.
With the use of the artificial insemination method, the offspring from individual stallions will amount to many tens and even hundreds, as a result of which the desired composition of stallions and mares will be more numerous, more uniform and consistent in quality. Breeding work with such a composition will be easier and more successful, especially since the method of artificial insemination should be widely used mainly by those stallions that consistently and fully pass on their valuable qualities to the offspring of various mares.
The danger of closely related breeding can be easily eliminated by replacing some stallions with others (as is practiced in sheep farming in relation to rams) every 3-5 years, which does not present any particular difficulties in the conditions of socialist horse breeding.
Transfers and replacements of stud stallions, which are inevitable even with the natural mating of horses, should be more frequent with artificial insemination.
Artificial insemination in horse breeding practice will make it possible to widely use the most valuable producers and apply the linear breeding method with maximum efficiency. Dissemination of the most valuable qualities of the ancestors and successors of the lines is a powerful lever for the rapid improvement of the breed. An opportunity is being created to abandon the use of secondary manufacturers.
Artificial insemination opens up broad prospects for the qualitative improvement of horses because it makes it possible to inseminate the semen of outstanding mares, which during natural mating are usually assigned only to mediocre stallions.
Artificial insemination does not lead to a reduction in the lines with which breeding work is carried out, but contributes to the displacement of secondary successors of these lines that are forced to be used in natural mating.
Artificial insemination makes it possible to more accurately, quickly and completely evaluate breeding stallions based on the quality of their offspring than during natural mating of horses.
Some horse breeding workers believe that artificial insemination of horses should be introduced primarily in the collective farm sector in order to quickly improve the bulk of the horse population.
We cannot completely agree with this opinion. If you don’t engage in artificial insemination in horse breeding, then where can you get good breeding stallions-improvers for artificial insemination centers?
The introduction of artificial insemination into widespread horse breeding collective farm practice without a sufficient number of good stallions will necessitate the use of mediocre and even unsatisfactory stallions.
Artificial insemination of mares with the semen of mediocre stallions has no production value, and insemination with the semen of unsatisfactory stallions can bring nothing but harm and should be strictly prohibited.
Stud farms generally have better conditions (for example, availability of qualified livestock specialists and veterinarians) for organizing and conducting artificial insemination of horses than the mass horse breeding sector.
We believe that artificial insemination of horses should be introduced primarily into breeding stud farms and large horse-breeding collective farms that have good stallions and are serviced by qualified
Zootechnical and veterinary personnel. In the mass sector of horse breeding, artificial insemination points should be organized only where there are the necessary conditions for this, in particular, good class I stud stallions.
The network of these points in the mass horse breeding sector should expand depending on their replenishment from breeding stud farms and nurseries with stallions suitable for widespread use by artificial insemination.
When grading stallions, you need to make a mark on the grading card with a precise indication of whether a given stallion is suitable or unsuitable for widespread use by artificial insemination.
Not every stallion, even an elite one, is desirable for this purpose.
The method of artificial insemination of horses is unthinkable without breeding zootechnical selection and should not be used without it.