CN102986647B - Non-animal-origin frozen semen diluent - Google Patents
Non-animal-origin frozen semen diluent Download PDFInfo
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- CN102986647B CN102986647B CN201110264510.6A CN201110264510A CN102986647B CN 102986647 B CN102986647 B CN 102986647B CN 201110264510 A CN201110264510 A CN 201110264510A CN 102986647 B CN102986647 B CN 102986647B
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Abstract
The invention relates to a non-animal-origin frozen semen diluent, and relates to a frozen semen diluent prepared with non animal-origin component. The diluent is used for preparing cattle and sheep frozen semen. In prior arts, adopted frozen semen contains animal-origin components, and prepared frozen semen has the defects of low motility and short survival time, low intact acrosome rate of thin tube frozen semen, and large amount of pathogenic micro-organisms after thawing. With the diluent provided by the invention, the problems are solved. The diluent is prepared from the components of, by mass, 3.412 g of lactose, 0.5 g of sucrose, 0.9 g of glucose, 0.61 g of trehalose, 0.53 g of citric acid, 0.403 g of acetic acid tetraethylammonium, 1.425 g of tris(hydroxymethyl) aminomethanegrams, 0.375 g of tri-sodium citrate, 5.2 ml of ethylene glycol, 100 ml of double-distilled water, and 0.88 g of phospholipids powder. With the diluent, prepared frozen semen has the advantages of high motility and long survival time, high intact acrosome rate of thin tube frozen semen, and small amount of pathogenic micro-organisms after thawing.
Description
Technical field:
The present invention relates to a kind of extender of freezing semen with the preparation of animal origin-free composition using when ox, sheep are frozen essence of making.
Background technology:
At present, when making ox, sheep are frozen essence, need to seminal fluid, dilute with dilution, and the extender of freezing semen using at present, all contain animal origin composition (containing lecithality, milk etc.), with its making freeze essence thaw after sperm viability low, the time-to-live is short, the acrosomal integrity of straw frozen semen is low, and sex pheromone quantity is many.Under the prerequisite that is 70% at former smart vigor, freeze essence thaw after sperm viability 30～50%, average vigor only has 45%, at the mean survival times of 38 ℃, it is 4～7 hours, after thawing, the acrosomal integrity of straw frozen semen is on average below 65%, and after within 24 hours, cultivating, the quantity of sex pheromone is on average at 600～1200 clump counts.
Summary of the invention:
Technical problem to be solved by this invention is to provide a kind of non-animal derived extender of freezing semen, of this dilution, make freeze essence thaw after sperm viability high, the time-to-live is long, the acrosomal integrity of straw frozen semen is high, sex pheromone quantity is few.
Above-mentioned purpose is achieved in that described non-animal derived extender of freezing semen is formulated by following mass ratio by following composition: 3.412 grams of lactose, 0.5 gram of sucrose, 0.9 gram of glucose, 0.61 gram of trehalose, 0.53 gram of citric acid, 1.425 grams, 0.403 gram of tri-Hydroxyalkyl methylamino methane of ethanedioic acid triethylammonium tetrakis, 0.375 gram of trisodium citrate, 5.2 milliliters of ethylene glycol, 100 milliliters of distilled waters, powder phosphatidase 10 .88 gram.
Concrete compound method is as follows: by described lactose, sucrose, glucose, trehalose, citric acid, ethanedioic acid triethylammonium tetrakis, tri-Hydroxyalkyl methylamino methane, trisodium citrate, ethylene glycol, put into conical flask, add distilled water, be heated to 95 ℃, above-mentioned medicine dissolved completely, while being cooled to 35 ℃, add powder phosphatide, stir 30 minutes, then be cooled to 4 ℃, and in 4 ℃ of environment standing depositing 12 hours, sediment with in filter paper filtering solution, can be used.
Advantage of the present invention is: of this dilution, make freeze essence thaw after sperm viability high, the time-to-live is long, the acrosomal integrity of straw frozen semen is high, sex pheromone quantity is few.Under the prerequisite that is 70% at former smart vigor, the average Frozen semen activity of separating is more than 58%, the highest vigor of thawing reaches 68%, after thawing in 38 ℃ of environment the time-to-live on average more than 11 hours, the live time that lives forever was most over 24 hours, after thawing, the average acrosomal integrity of straw frozen semen is more than 72.8%, the essence of freezing after thawing was cultivated through 48 hours, sex pheromone clump count is less than 130, if guaranteed during semen collection in former essence and operating process without pathogeny microbial contamination, the straw frozen semen of making does not just have sex pheromone and exists after cultivating; Apply the made essence of freezing of this dilution, can avoid because of dilution with sex pheromone between cattle and sheep, mutually propagate between cattle and sheep and people, thereby effectively reduce the risk of infecting both domestic animals and human epidemic disease.
Embodiment:
Described non-animal derived extender of freezing semen is formulated by following mass ratio by following composition: 3.412 grams of lactose, 0.5 gram of sucrose, 0.9 gram of glucose, 0.61 gram of trehalose, 0.53 gram of citric acid, 1.425 grams, 0.403 gram of tri-Hydroxyalkyl methylamino methane of ethanedioic acid triethylammonium tetrakis, 0.375 gram of trisodium citrate, 5.2 milliliters of ethylene glycol, 100 milliliters of distilled waters, powder phosphatidase 10 .88 gram;
Concrete compound method is as follows: by described lactose, sucrose, glucose, trehalose, citric acid, ethanedioic acid triethylammonium tetrakis, tri-Hydroxyalkyl methylamino methane, trisodium citrate, ethylene glycol, put into conical flask, add distilled water, be heated to 95 ℃, above-mentioned medicine dissolved completely, while being cooled to 35 ℃, add powder phosphatide, stir 30 minutes, then be cooled to 4 ℃, and in 4 ℃ of environment standing depositing 12 hours, sediment with in filter paper filtering solution, can be used.
Wherein lactose, sucrose, glucose, trehalose, citric acid, ethanedioic acid triethylammonium tetrakis, tri-Hydroxyalkyl methylamino methane, trisodium citrate, ethylene glycol are chemical analysis pure reagent, and the pH value of distilled water is 6.9.
Wherein powder phosphatide refers to the soybean lecithin of being produced by the permanent company of Beijing Hua Qingmei, specific targets are as follows: product type is PLW, mode of appearance is light yellow powder, moisture content and volatile matter≤1.0%, acetone insoluble matter >=98%, ether insoluble matter≤0.1%, acid value (mgKOH/g)≤35, peroxide value≤12.
Wherein trisodium citrate is the trisodium citrate that contains two water of crystallization.
With this non-animal derived extender of freezing semen, make and freeze smart method:
1. sperm dilution:
Before use, the non-animal derived extender of freezing semen water-bath preparing is heated to 35 ℃, then the seminal fluid and the non-animal derived extender of freezing semen that gather were diluted by 1: 2～1: 15.
Cooling balance:
The seminal fluid having diluted is cooled to 0～5 ℃ with the speed of approximately 0.5 ℃ per minute, deposits 0.5 hour, then at this temperature, carry out filling.
3. freezing:
Filling good straw frozen semen is positioned over and at the temperature of-95 ± 5 ℃, carries out freezingly, within 3 minutes, by detecting in qualified input liquid nitrogen, preserve.
4. straw frozen semen thaws:
From liquid nitrogen, take out straw frozen semen, put into rapidly in the warm water of 38～40 ℃, make water surface submergence freeze fine tube, to straw frozen semen, just become when transparent and taken out, at the micro-Microscopic observation of 200～600 times.With this non-animal derived extender of freezing semen, make the breeding effect after straw frozen semen thaws:
Cattle and sheep adopt cervical orifice semen deposition method breeding in the onset of ovulation, feelings phase semen deposition 2 times, and 12 hours, interval, the effect of being impregnated is: ox average out to is more than 97%, and sheep is more than 76%.The conception rate of ox on average improves 5～7% than the conception rate of the made frozen semen of dilution with containing animal origin, and the conception rate of sheep on average improves 10～16% than the conception rate of the made frozen semen of dilution with containing animal origin.
Claims (1)
1. non-animal derived extender of freezing semen, it is characterized in that by following composition formulated by following mass ratio: 3.412 grams of lactose, 0.5 gram of sucrose, 0.9 gram of glucose, 0.61 gram of trehalose, 0.53 gram of citric acid, 1.425 grams, 0.403 gram of ， tri-Hydroxyalkyl methylamino methane of ethanedioic acid triethylammonium tetrakis, 0.375 gram of trisodium citrate, 5.2 milliliters of ethylene glycol, 100 milliliters of distilled waters, powder phosphatidase 10 .88 gram.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
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| CN201110264510.6A CN102986647B (en) | 2011-09-08 | 2011-09-08 | Non-animal-origin frozen semen diluent |
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| Application Number | Priority Date | Filing Date | Title |
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| CN201110264510.6A CN102986647B (en) | 2011-09-08 | 2011-09-08 | Non-animal-origin frozen semen diluent |
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| CN102986647A CN102986647A (en) | 2013-03-27 |
| CN102986647B true CN102986647B (en) | 2014-04-02 |
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| CN201110264510.6A Expired - Fee Related CN102986647B (en) | 2011-09-08 | 2011-09-08 | Non-animal-origin frozen semen diluent |
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Families Citing this family (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2015193265A1 (en) | 2014-06-16 | 2015-12-23 | Università degli Studi di Parma | Composition for extenders for the long-term conservation of animal seminal material |
| CN111066775B (en) * | 2016-06-29 | 2022-02-01 | 许红喜 | Method for cold storage of animal sperm |
| CN109907038B (en) * | 2019-04-30 | 2019-12-31 | 黑龙江省农业科学院畜牧兽医分院 | Animal-origin-free bovine frozen semen diluent |
| CN110301431A (en) * | 2019-07-17 | 2019-10-08 | 西北民族大学 | A kind of non-animal derived property sheep sperm cryo-conservation dilution and method |
| CN110367240A (en) * | 2019-07-17 | 2019-10-25 | 西北民族大学 | A kind of sheep sperm cryo-conservation dilution and method |
| CN113615682B (en) * | 2021-09-05 | 2022-08-26 | 内蒙古赛诺种羊科技有限公司 | Sheep hypertonic complexing agent semen dilution preserving fluid and application method thereof |
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| EP1051907A1 (en) * | 1999-05-14 | 2000-11-15 | IMV Technologies | Extender for cryopreserving bovine sperm |
| CN101220345A (en) * | 2008-01-23 | 2008-07-16 | 西北农林科技大学 | Cattle freezing seminal fluid dilution and method for producing the same |
| CN101310729A (en) * | 2007-05-23 | 2008-11-26 | 辽宁省辽宁绒山羊育种中心 | Animal origin free flocks and herds frozen semen diluent and production method of flocks and herds frozen semen |
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- 2011-09-08 CN CN201110264510.6A patent/CN102986647B/en not_active Expired - Fee Related
Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| EP1051907A1 (en) * | 1999-05-14 | 2000-11-15 | IMV Technologies | Extender for cryopreserving bovine sperm |
| CN101310729A (en) * | 2007-05-23 | 2008-11-26 | 辽宁省辽宁绒山羊育种中心 | Animal origin free flocks and herds frozen semen diluent and production method of flocks and herds frozen semen |
| CN101220345A (en) * | 2008-01-23 | 2008-07-16 | 西北农林科技大学 | Cattle freezing seminal fluid dilution and method for producing the same |
Non-Patent Citations (4)
| Title |
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| 卵磷脂取代卵黄冷冻牛精液效果观察;胡传活 等;《江苏农业科学》;20061231(第6期);第285页左栏1.1 主要仪器设备及试剂，第287页左栏第2段 * |
| 去精清对提高辽宁绒山羊细管冻精质量的影响;宋先忱 等;《辽宁畜牧兽医》;20040831(第8期);第18页左栏1.2 稀释液配方 * |
| 宋先忱 等.去精清对提高辽宁绒山羊细管冻精质量的影响.《辽宁畜牧兽医》.2004,(第8期), |
| 胡传活 等.卵磷脂取代卵黄冷冻牛精液效果观察.《江苏农业科学》.2006,(第6期), |
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