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31For citation: Baldan T., Myagmardulam P., Ankhbayar T., Jambatsadmid D., Khosbayar B. Research results on the use of frozen yak sperm //
URL: http://rectors.altstu.rU/ru/periodical/archiv/2022/1/articles/4_2.pdf DOI: 10.25712/ASTU.2410-485X.2022.01.008 EDN: https://elibrary.ru/sjgujn
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Research results on the use of frozen yak sperm
T. Baldan1, P. Myagmardulam1, T. Ankhbayar2, D. Jambatsadmid2, B. Khosbayar2
1 Mongolian University of Life Science,
2 Mongolian livestock Genefound Center
Historical background
Mongolian yak is one of the unique populations of Central Asian highland alpine pastures and climates, adapted to the specifics of the climate.
The number of yaks is increasing year by year, but it is losing its majestic appearance, its body is getting better, and its productivity is declining.
This is due, firstly, to the fact that over the past two decades, Mongolian yak has lead to declining to bred and selection work and secondly, to climate change and global warming to some extent affecting the yak's body.
Researcher Yo. Zagdsuren for the first time comprehensively studied the phenomenon of heterosis not only from the economic point of view but also from the biochemical, physiological, and immunogenetic point of view.
The khainag have been found to be significantly higher than the average of yak and cattle species in terms of growth rate and several biochemical parameters.
Heterosis is caused by the activation of various enzymes that stimulate the growth and development of metabolism, which is explained by the fact that the combination of genes in Mongolian cattle and yaks produces enzymes with hybrid molecules.
Researchers in some foreign yak breeding countries have done a lot of research. For example, B.C. In 2004, Das and M. Sarkar organized a training course in India called "Training and collection of semen from yak bull (Poephagus grunniens L.). They also published the effect of thawing methods on frozen semen quality of yak (Poephagus grunniens L.) bulls.
Cygnaan Binod Kumar Dutta Borah, Bharat Chandra Deka, Ranjan Kumar Biswas, Prithiviraj Chakravarty, Sourabh Deori, Sudip Sinha, and Kutubuddin Ahmed Haptffl cyganraaHti a^nn 2015 нннтпэгg^ээ.
Researchers (Binod Kumar Dutta Borah, Bharat Chandra Deka, Ranjan Kumar Biswas Prithiviraj Chakravarty, Sourabh Deoril, Sudip Sinha, Kutubuddin Ahmed) were published an article about yak study in 2015.
Susana Campi, Andres Jorge, Daniel Lombardo, Carlos Blasi, Mariana Gambarotta, Maria Laura Fischman, Humberto Cisale conducted a joint study in 2016 and published Yak (Bos Grunniens) sperm nuclei morphology, morphometry, and DNA content.
Mongolian native yak
Biological characteristics:
Mongolian native yak is one of the most distinctive populations and well adapted in the taiga area of grassland and high mountain ranges in Central Asia, Mongolia.
The yaks have unique biological peculiarity to withstand with naturally and climatically severe harsh conditions. They can walk easy on any mountainous obstacles such as rapid ascent, downhill, steep slope and loose rock, rocky and use sparse bush, grasses, woody shrubs and thorns. The yak has long, coarse hair all over its body especially on shoulder, hip and belly acting as insulation when lying in cold and snow covered places.
They have a long bushy tail and an extremely long hair from belly to ankle. Long, thick hair protects them from cold and heat as well as providing insulation when lying in cold and snowy places. In addition, the animal grows dense undercoat for cold protection. Mongolian local yak has poll, distance between which is shorter, short and thick neck, well developed brisket, withers higher, lower withers, deep chest, straight back, diagonal body-short, sloppy and narrow croup and hind limb gait is sometimes closer between hocks, end body mostly slim. The Mongolian khainag has biggest body, chest bigger, body longer, short lag, with precipitous and rampage.
In mainly yak populations are polled with the 75-85% of total yaks. 63.2 percent of dominant color yak is black, dark, black brown and 15.2%-blue and 12.4%-white and light white color.
Productive parameters (tab.1)
Age, sex body weight, milk quality
kg Output, liter fat, %
yak bull 400-450 - -
yak cow 270-280 280-320 7.2-12.0
Mineral component of yak, khainag and Mongolian native cow's milk (tab.2)
type Ash, % calcium, (Ca) mg/% phosphor (P), mg /%
Yak 0.89 130.6 106.22
khainag (hybrid) 0.93 134.0 134.82
Mongol cow 0.88 124.91 97.82
Productivity: Yak and khainag's milk, meat are used for food and skin, fiber is raw material to industry. And they are used for nomadic transport, riding. Carcass yield of adult castrated yak is 52.2 percent and the fat of the yak meat will be less accumulated to the muscles and meat fat is higher accumulated in internal organ.
Calving mostly occurs between March and June. Milk production depends on the lactation period. Yak cow's milk production average year is 563-738 liter; milk fat rate is 6.7-8.9%, protein — 5.31%, dry matter-18.71%, 5.2% lactose. Mongolian local yak's milk consist bigger dry matter, fat, protein, lactose than comparison to other breed cows because it is making product is very important product.
Yak and khainag's milk are suitable for making product cheese, yogurt, dry yogurt, curds because, these milk is biggest by protein and its protein is a good coagulated and casein.
Distribution: Mongolian local yak have been breeding in high mountainous area of Arkhangai, Khuvsugul, Bayankhongor, Uvurkhangai, Zavkhan, Gobi-Altai, Khovd, Bayan-Ulgii provences. Yak and Khainag (hybrid of Yak and Mongolian native cattle) occurred 25% of total Mongolian cattle. Yak bred in mountainous area with 2000 meter above sea level of Mongolia, Tuvd, Balba of India and Gorno-Altai of Russia, Kyrgyz Republic. In census 2017, 792.2 thousand head of yak and khainag were inventoried in Mongolia.
Materials and methods
This study was conducted in September, October 2019 at the Semen Laboratory of the National Genebank Center of Livestock.
Yak bull selection: The 3 younger and 2 old yak bull was selected based on general body development, weight, exterior, body shape, color, hair cover, long bushy tail, body condition, fertility and temperament from Ikhtamir soum of Arkhangai aimag. And selected one head nonpregnant young cow with calf.
Adapting for supplementary feed: In collaboration with specialists from the National Genebank Center for Livestock, bulls were supplemented with high-quality green grass and forage from 25 August.
For the first 10 days, the bulls and cows were provided with good quality green grass and to take semen collect in AI vagine.
Green grass forage with egg and carrots
Semen collection from yak bull
Between September 26 and October 2, we collected semen from yak bulls. And kept wich deep-frozen semen.
preparation work for semen collection
Estrouse synchronization of yak cows: A combination of prostaglandin F2a and progesterone was injected to synchronize of yak cow with nonpregnant.
Hormonal combinations were used in several different variants for the cow's synchronization, but the yak cow's estrous was badly detected. However, some of the bulls were mounted on this cow, so detected estrous that the bed was them.Three dairy cows were selected for synchronizing and those of the three cows estrous detected were used for semen collection of yak bull.
Semen collection of yak bull
In all animal husbandry countries, artificial vaginas are used to collect semen from a variety of animals. The artificial vagina provides the same conditions as the vagina's heat, pressure, and slipperiness in breeding animals.
For many days, those yak bulls were bred together with dairy cows, so horned brown bulls and dehorned white bulls gave the semen to vagine.
Yak bull mounted on dairy cattle cow
Semen collecting process
Semen dilution
The Andromeda diluent supplied by Minitube was used to dilute the yak semen.
Evaluate semen quality
Semen quality was determined by its color, odor, volume, sperm motility, number of living and dead sperm, number of sperm per ml of sperm, and number of defected sperm.
Freezing and keeping
The yak's semen was evaluated by the center's Molecular Biology and Genetics Laboratory and deep-frozen using the latest equipment and technology supplied by the world-famous Mini tube company. Using a spectrophotometer, determined the volume of the yak bull semen, the degree of dilution, and the number of straws required.
Straw coding and semen packing
The spectrophotometer determined the number of doses and encoded the straws. The following markings were made on a single-dose straw. These include: breed name, yak bull name, tag number, date of seed production, packer number.
Semen straw cod marked straw
The yak bull semen is packed automatically using special device equipment.
Semen freezing to higher temperature
It is estimated that the number of active sperm in a single-dose seed should be at least 25 million after thawing of frozen seeds before deep-frozen storage. The diluted
semen is packaged in 0.25ml or 0.5ml straws to have the above-mentioned active sperm.
Before freezing, refrigerate at 2-4°C for 3-4 hours and allow the sperm to cool. It was then placed in a plastic box with a special tray, placed horizontally, in a pre-liquid nitrogen cuvette, touched to the bottom of the box, and frozen in vapor. Liquid nitrogen vapor in such an environment is -196°C, so the seeds freeze very quickly in 3-4 minutes.
Keeping frozen semen
Frozen semen was stored vertically in a Dewar container with liquid nitrogen at -196°C through a semen storage container.
Semen thawing
Deep-frozen seeds have thawed a straw in a pot of hot water at 38 to 40°C for about 15 sec. For artificial insemination of breeding animals, semen with a value of at least 3-4 points should be used after thawing. In order to assess the quality of the frozen semen in the straw, the seeds were thawed according to the technology.
Result of study
Semen collection of yak bull
The semen was collected from 5 yaks of different ages.
Result of Semen collected work (tab.3)
n bull name mounted giving semen volume (ml) explanation
1 Brown horned bull + + 6 semen froze
2 Dehoned white bull + + 1.0 small volume, poor quality
3 Bigger black bull 0 no mounted
4 Dehoned black bull 0 not gave semen for vagine
5 Dehoned black bull - _ 0 not gave semen for vagine
Result of synchronization for dairy cattle cows and yak cow: (tab.4)
n cow name breeder name injection date injection size (ml) estrous detect
1 Dehoned black heifer yak Prostaglandine (F2a) 4ml - 4 time detected
2 black cow dairy cattle Prostaglandine (F2a) 5 ml - 3 time detected
n cow name breeder name injection date injection size (ml) estrous detect
3 Brown heifer dairy cattle Prostaglandine (F2a) 5 ml detected
4 Red cow dairy cattle Prostaglandine (F2a) 5 ml detected
5 Grey heifer dairy cattle Prostaglandine (F2a) 5 ml no detected
80 percent of all cows detected estrous by injected prostaglandin hormone.
Semen volume and semen quality
In our study, the evaluation of 2.3 ml of yak bull semen was as follows.
Some index of yak bull semen: (tab.5)
bull name semen volume by one time mounted (ml) sperm motility (%) Sperm concentration (million)
Horned brown 2.3 83.95 1.240
The number of semen given in a single mount was 2.3 ml with relatively small but dense semen. In addition, the sperm activity is 83.95 percent and the number of sperm cells is 1 billion 240 million, which indicates that the seed is of good quality. When the quality of the horned bull's horns was evaluated using a spectrophotometer, the sperm activity was 83.95 percent and 1 billion 240 million sperm.
Sperm motility
The number, shape, and mobility of sperm are important for testing for male factor infertility.
Some index of yak bull semen (tab.6)
bull name number of normal sperm (%) Progressive motility (%) Slow motility (%) Abnormal Хэлбэлзэх, % dead sperms (%)
horned brown 87.93 64.22% 16.38 1.72 5.60 12.07
The table shows that the semen activity is 87.93 percent, and the forward-seeded seeds account for 64.22 percent of the total semen, indicating that the semen is of good quality. And, the percentage of dead sperms was 12.07 percent.
Frozed semen quality
Some index of frozen semen, one time: (tab.7)
name index
Volume (ml) 2.3
Concentration (%) 83.95
Dilution grade 20
Sperm total number (mln) 1240
Number of semen straws 184
Loss 15
Frosed semen 169
Quality of frozen semen (point) 3-5
During the study, 420 straws of yak bull semen were deep-frozen. Discussion
In 2004, B.C.Das and M. Sarkar conducted a study entitled "Preparing and Collecting Semen from Yak Bulls" to obtain and train yak bulls using artificial vaginas.
Dr. B.Erdenebaatar and Ch. Gombojav, researchers at the Research Institute of Animal Husbandry was attempted to artificially inseminate for Mongolian yak cow. The study, in the future, our research has shown that it is possible to protect the gene found in yak herds by using the yak bull semen freezing method.
In 2017, S.Deori has studied the effect of freezing on cryo survival of yak sperm in India. Yak semen hase a better survival while freezing in tris extender with 6.4percent glycerol and 20 percent egg yolk following and equilibration perion of 4h. Frozen yak semen can be suitably used artificial insemination to curb inbreeding.
Conclusion
1. During the study, 420 straws of yak bull semen were deep-frozen in the liquied nitrogen with 196 C degrees.
2. The horned-brown bull's sperm activity is 83.95 percent and the number of sperm cells is 1 billion 240 million.
3. Attempts were to collect semen from five yak bulls total and two bull gave semen, results is 40 percent. It was very difficult to collect semen in artificial vagine from Mongolian yak bull. This is due to the biological characteristics and innate behavior of yaks.
4. 80 percent of all cows detected estrous by injected prostaglandin hormone.
5. Deep-frozen semen activity is rated at 3-5 points, indicating that it can be used for artificial insemination in the future. In the future, our research has shown that it is possible to protect the gene found in yak herds by using the yak bull semen freezing method.
References
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