CN102986647A - Non-animal-origin frozen semen diluent - Google Patents
Non-animal-origin frozen semen diluent Download PDFInfo
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- CN102986647A CN102986647A CN2011102645106A CN201110264510A CN102986647A CN 102986647 A CN102986647 A CN 102986647A CN 2011102645106 A CN2011102645106 A CN 2011102645106A CN 201110264510 A CN201110264510 A CN 201110264510A CN 102986647 A CN102986647 A CN 102986647A
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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 of making ox, using when sheep is frozen essence.
Background technology:
At present, the making ox, when sheep is frozen essence, need with dilution seminal fluid to be diluted, and the extender of freezing semen that uses at present, all contain animal origin composition (containing lecithality, milk etc.), with its making to freeze the essence rear sperm viability of thawing low, the time-to-live is short, the acrosomal integrity of straw frozen semen is low, and sex pheromone quantity is many.Be under 70% the prerequisite at former smart vigor, freeze essence and thaw rear sperm viability 30～50%, average vigor only has 45%, it is 4～7 hours at 38 ℃ mean survival times, thaw the acrosomal integrity of rear straw frozen semen on average below 65%, and the quantity of sex pheromone is on average at 600～1200 clump counts after cultivating in 24 hours.
Summary of the invention:
Technical problem to be solved by this invention provides a kind of non-animal derived extender of freezing semen, with this dilution make to freeze the essence rear sperm viability of thawing high, the time-to-live is long, the acrosomal integrity height of straw frozen semen, 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: lactose 3.412 grams, sucrose 0.5 gram, glucose 0.9 gram, trehalose 0.61 gram, citric acid 0.53 gram, ethanedioic acid triethylammonium tetrakis 0.403 gram San Hydroxyalkyl methylamino methane 1.425 grams, trisodium citrate 0.375 gram, 5.2 milliliters of ethylene glycol, 100 milliliters of distilled waters, powder phosphatidase 10 .88 gram.
Concrete compound method is as follows: with described lactose, sucrose, glucose, trehalose, citric acid, ethanedioic acid triethylammonium tetrakis, San Hydroxyalkyl methylamino methane, trisodium citrate, ethylene glycol, put into conical flask, add distilled water, be heated to 95 ℃, above-mentioned medicine dissolved fully, when being cooled to 35 ℃, add powder phosphatide, stirred 30 minutes, and be cooled to again 4 ℃, and in 4 ℃ of environment, leave standstill and deposit 12 hours, sediment with in the filter paper filtering solution can use.
Advantage of the present invention is: with this dilution make to freeze the essence rear sperm viability of thawing high, the time-to-live is long, the acrosomal integrity height of straw frozen semen, sex pheromone quantity is few.Be under 70% the prerequisite at former smart vigor, on average thaw rear vigor more than 58%, the highest vigor of thawing reaches 68%, thaw after in 38 ℃ of environment the time-to-live on average more than 11 hours, the live time that lives forever most was above 24 hours, thaw the average acrosomal integrity of rear straw frozen semen more than 72.8%, freezing after thawing is smart in cultivation in 48 hours, the sex pheromone clump count is less than 130, if guarantee during semen collection in former essence and the operating process without pathogeny microbial contamination, the straw frozen semen of then making does not just have sex pheromone and exists after cultivating; Use the made essence of freezing of this dilution, can avoid because of dilution with sex pheromone between the cattle and sheep, mutually propagate between cattle and sheep and the 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: lactose 3.412 grams, sucrose 0.5 gram, glucose 0.9 gram, trehalose 0.61 gram, citric acid 0.53 gram, ethanedioic acid triethylammonium tetrakis 0.403 gram San Hydroxyalkyl methylamino methane 1.425 grams, trisodium citrate 0.375 gram, 5.2 milliliters of ethylene glycol, 100 milliliters of distilled waters, powder phosphatidase 10 .88 gram;
Concrete compound method is as follows: with described lactose, sucrose, glucose, trehalose, citric acid, ethanedioic acid triethylammonium tetrakis, San Hydroxyalkyl methylamino methane, trisodium citrate, ethylene glycol, put into conical flask, add distilled water, be heated to 95 ℃, above-mentioned medicine dissolved fully, when being cooled to 35 ℃, add powder phosphatide, stirred 30 minutes, and be cooled to again 4 ℃, and in 4 ℃ of environment, leave standstill and deposit 12 hours, sediment with in the filter paper filtering solution can use.
Wherein lactose, sucrose, glucose, trehalose, citric acid, ethanedioic acid triethylammonium tetrakis, San Hydroxyalkyl methylamino methane, trisodium citrate, ethylene glycol are the chemical analysis pure reagent, and the pH value of distilled water is 6.9.
Wherein powder phosphatide refers to the soybean lecithin by the production of 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.
Make the method for freezing essence with this non-animal derived extender of freezing semen:
1. sperm dilution:
Before use, the non-animal derived extender of freezing semen water-bath for preparing is heated to 35 ℃, again seminal fluid and the non-animal derived extender of freezing semen that gathers diluted by 1: 2～1: 15.
2. cooling balance:
The seminal fluid that dilution is good is cooled to 0～5 ℃ with the about 0.5 ℃ speed of per minute, deposits 0.5 hour, then carries out can under this temperature.
3. freezing:
The straw frozen semen that can is good is positioned over and carries out freezingly under-95 ± 5 ℃ the temperature, preserves by detecting in the qualified input liquid nitrogen in 3 minutes.
4. straw frozen semen thaws:
From liquid nitrogen, take out straw frozen semen, put into rapidly in 38～40 ℃ the warm water, make water surface submergence freeze fine tube, just become to straw frozen semen and taken out when transparent, observe at 200～600 times microscopicallies.Make breeding effect after straw frozen semen thaws with this non-animal derived extender of freezing semen:
Cattle and sheep are adopted cervical orifice semen deposition method breeding in the onset of ovulation, feelings phase semen deposition 2 times, and interval 12 hours, the effect of being impregnated is: the 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 with the made frozen semen of dilution that contains animal origin, and the conception rate of sheep on average improves 10～16% than the conception rate with the made frozen semen of dilution that contains 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: lactose 3.412 grams, sucrose 0.5 gram, glucose 0.9 gram, trehalose 0.61 gram, citric acid 0.53 gram, ethanedioic acid triethylammonium tetrakis 0.403 gram ， San Hydroxyalkyl methylamino methane 1.425 grams, trisodium citrate 0.375 gram, 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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| Publication Number | Publication Date |
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| CN102986647A true CN102986647A (en) | 2013-03-27 |
| CN102986647B 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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Cited By (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 |
| CN109907038A (en) * | 2019-04-30 | 2019-06-21 | 刘学峰 | A kind of non-animal derived ingredient frozen cattle semens dilution |
| 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 |
| CN110622958A (en) * | 2016-06-29 | 2019-12-31 | 许红喜 | Animal sperm diluent |
| CN113615682A (en) * | 2021-09-05 | 2021-11-09 | 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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| 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
- 2011-09-08 CN CN201110264510.6A patent/CN102986647B/en not_active Expired - Fee Related
Patent Citations (3)
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| 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 |
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| 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》, no. 8, 31 August 2004 (2004-08-31) * |
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Cited By (8)
| 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 |
| CN110622958A (en) * | 2016-06-29 | 2019-12-31 | 许红喜 | Animal sperm diluent |
| CN110622958B (en) * | 2016-06-29 | 2022-02-01 | 许红喜 | Animal sperm diluent |
| CN109907038A (en) * | 2019-04-30 | 2019-06-21 | 刘学峰 | A kind of non-animal derived ingredient frozen cattle semens dilution |
| 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 |
| CN113615682A (en) * | 2021-09-05 | 2021-11-09 | 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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| Publication number | Publication date |
|---|---|
| CN102986647B (en) | 2014-04-02 |
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