CN101200688A - Boar semen freezing preservation tube and method thereof - Google Patents
Boar semen freezing preservation tube and method thereof Download PDFInfo
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- CN101200688A CN101200688A CNA2007101563988A CN200710156398A CN101200688A CN 101200688 A CN101200688 A CN 101200688A CN A2007101563988 A CNA2007101563988 A CN A2007101563988A CN 200710156398 A CN200710156398 A CN 200710156398A CN 101200688 A CN101200688 A CN 101200688A
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- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61D—VETERINARY INSTRUMENTS, IMPLEMENTS, TOOLS, OR METHODS
- A61D19/00—Instruments or methods for reproduction or fertilisation
- A61D19/02—Instruments or methods for reproduction or fertilisation for artificial insemination
- A61D19/022—Containers for animal semen, e.g. pouches or vials ; Methods or apparatus for treating or handling animal semen containers, e.g. filling or closing
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Abstract
The present invention relates to a porcine semen freezing and storing tube and a porcine semen freezing and storing method thereof. The porcine semen freezing and storing tube comprises a storing tube and is characterized in that the storing tube is a double-layer tube; the two vessel openings of the double-layer tube are opposite. The key points of the porcine semen freezing and storing method are that the frozen and diluted porcine semen after the pretreatment and the cooling treatment is cased into the annular vessel of the porcine semen freezing and storing tube of the present invention; then the porcine semen freezing and storing tube is inserted at liquid nitrogen to be suffocated for a plurality of minutes and directly put into the liquid nitrogen for long time storage, which can increase the contacting area of the semen and the liquid nitrogen during the process of freezing, accelerate semen freezing velocity, reduce the damage to the semen caused by freezing and improve the live-rate of frozen semen. The present invention also has the advantages of low cost, convenient operation, high efficiency, ensuring fecundation rate, etc.
Description
Technical field
The present invention relates to a kind of Cryopreservation of Boar Semen pipe and Cryopreservation of Boar Semen method thereof.
Background technology
In China, pig is topmost feeding stock animals, and pork output accounts for 70% of national meat total amount.Along with the raising of living standards of the people, the domestic and international market is transformed into quality (as meat etc.) from quantity gradually to the requirement of market pig.The breeding direction of pig also from mainly being conceived to the raising of the speed of growth and lean ratio, changing into improve pork quality, improves the local flavor of meat.Traditional seed selection mode time generation interval is long, can not meeting the market requirement, and adopt pig frozen semen technology, can make full use of good breeding stock, shorten time fine-variety breeding generation interval, set up the boar and the market pig cross breeding system of new suitable market demands fast.Simultaneously, the raising history of nearly one thousand years, hundreds of local variety that differ from one another have been formed in China, this is a very valuable property, but the introduction of external pig kind hybridization in recent years, made China that the extinction of a considerable amount of local variety is arranged or faced extinction, can effectively preserve China's local pig breed matter resource, have immeasurable social effect and benefit and adopt pig seminal fluid and embryo cryopreservation to preserve technology.
Though the freezing preservation technology of herbvore livestock semen very low temperature (196 ℃) is being brought into play important effect in modernized herding production, germ plasm resource in preserving.But the pig frozen semen is difficult to apply on a large scale aborning always, its reason has two, the one, C/P ratio significantly is lower than phytophagous animal in the pig spermatid plasma membrane, the ion permeability of sperm membrane strengthens in refrigerating process, lipid material runs off, and causes freezing that smart motility rate is low, conception rate is low.The 2nd, the semen volume that pig once breeds big (being about 50 milliliters), use traditional frozen semen formulation such as ampoule (about 1～2 milliliter), particle (about 0.5 milliliter), tubule (about 0.25 milliliter), all need the thaw essence of freezing of a great deal of (about 10～20) of each breeding, not only operate loaded down with trivial details, and the fertilization effect also be greatly affected.
Summary of the invention
At above problem, the purpose of this invention is to provide a kind of Cryopreservation of Boar Semen pipe and Cryopreservation of Boar Semen method thereof that can improve cold smart motility rate.
A kind of Cryopreservation of Boar Semen pipe comprises and preserves pipe, it is characterized in that: preserve pipe and be double wall tube, two vessel ports of double wall tube are opposite.Double wall tube of the present invention is meant the pipe that also is provided with a circle annular container outside the central, cylindrical container.
A kind of Cryopreservation of Boar Semen method comprises:
A, semen collection and with one times of isothermal BTS (Beltsville thawing solution) diluted,
B, under 15 ℃ of conditions balance 2 hours, 1000 left the heart 10 minutes then,
Seminal plasma is removed in c, centrifugal back, and the sperm precipitation is with cooling off diluted, and making sperm concentration is 2 * 10
9Individual/milliliter.Seminal fluid after dilution balance 2 hours under 5 ℃ of conditions,
D and then with freeze-extender 1: 1 dilution, the final densities that makes sperm is 1 * 10
9It is individual/milliliter,
It is characterized in that: the seminal fluid after the chill back of above step process is packed in the annular container of Cryopreservation of Boar Semen pipe of the present invention, then the Cryopreservation of Boar Semen pipe is inserted on the liquid nitrogen, stifling after several minutes, drop into directly that liquid nitrogen is medium-term and long-term to be preserved.
The present invention is by utilizing the opposite double wall tube of two vessel ports, is provided with also outside the central, cylindrical container promptly that the double wall tube of the opposite annular container of a circle opening realizes.So, in refrigerating process, can increase the contact area of seminal fluid and liquid nitrogen, accelerate semen freezing speed, reduce freezing damage to sperm, improve cold smart motility rate; It is low also to have cost, the operation box lunch, and the effect height guarantees advantages such as conception rate.
Description of drawings
Fig. 1 is a kind of Cryopreservation of Boar Semen tubular construction synoptic diagram of the present invention,
Fig. 2 is the right view of Fig. 1.
Among the figure: body 1, annular container 2, cylindrical container 3.
Embodiment
Describe in detail below in conjunction with drawings and Examples:
Cryopreservation of Boar Semen pipe among the figure is cylindrical double wall tube, and promptly double wall tube is a cylindrical tube.During use,, and double wall tube is inserted in the liquid nitrogen as long as the pig seminal fluid that extracts is packed in the annular container 2 that double wall tube forms, just can increase the contact area of seminal fluid and liquid nitrogen in the refrigerating process, accelerate semen freezing speed, reduce freezing damage, improve cold smart motility rate sperm.For the ease of making, reduce cost, double wall tube is made with plastic material.In order to satisfy artificial insemination dosage of pig, the number of times that simplifies the operation to greatest extent improves effect, reduces cost of labor, guarantees conception rate, and the volume of the annular container 2 of double wall tube is 5～15ml.
Method embodiment of the present invention is as follows:
1, pig seminal fluid pre-treatment.Select reproductivity height, athletic Jinhua herd boar, adopt the hand grip semen collection.Collect sperm and enrich part, after 4 layers of sterile gauze filter the removal colloid, survey semen volume immediately, and check sperm motility rate, about 150 milliliters at the seminal fluid more than 75% of collection sperm motility rates are with one times of isothermal BTS (Beltsville thawing solution) diluted.Balance is 2 hours under 15 ℃ of conditions, 1000 leaves the heart 10 minutes then.
2, seminal fluid cooling process.Seminal plasma is removed in the centrifugal back of pretreated pig seminal fluid, and the sperm precipitation is with cooling off diluted, and making sperm concentration is 2 * 10
9Individual/milliliter.Seminal fluid after dilution balance 2 hours under 5 ℃ of conditions.The prescription of cooling diluent is to contain lactose and 20 milliliters of yolk of 80 milliliter 11% in 100 milliliters of diluents.
3, semen freezing is handled.Seminal fluid after the cooling process is used freeze-extender dilution in 1: 1 again, and the final densities that makes sperm is 1 * 10
9Individual/milliliter.Then seminal fluid is packed in the annular container (2) of Cryopreservation of Boar Semen pipe of the present invention, every pipe is adorned 5～10 milliliters, places on the liquid nitrogen surface 3 centimetres, and is stifling after 20 minutes, drops into directly that liquid nitrogen is medium-term and long-term to be preserved.
In sum, the main points of the inventive method are: the seminal fluid after the chill back after pre-treatment of pig seminal fluid and the seminal fluid cooling process is packed in the annular container of Cryopreservation of Boar Semen pipe of the present invention, then the Cryopreservation of Boar Semen pipe is inserted on the liquid nitrogen, stifling after several minutes, directly drop into the medium-term and long-term preservation of liquid nitrogen.
In order to detect effect of the present invention, Jinhua, Zhejiang Province kind pig farm has been extracted successively the pig seminal fluid preserved according to the step process of the embodiment of the invention one and conventional process and has been kept at each 0.25 milliliter of pig seminal fluid in the straw, place 52 ℃ of water-baths to thaw in 20 seconds, detect the energy vigor at microscopically, wherein the present invention handles that sperm motility rate is 37.1% behind the pig semen thawing of preservation, and straw handles that motility rate is 35.9% behind the pig semen thawing of preserving.
In order to satisfy artificial insemination dosage of pig, every Cryopreservation of Boar Semen pipe is adorned 5～15 milliliters of pig seminal fluid after the chill back.Handle for improving preservation effect, the Cryopreservation of Boar Semen pipe that chill back pig seminal fluid is housed is inserted on the liquid nitrogen surface 3 centimetres, stifling after 20 minutes, drop into directly again that liquid nitrogen is medium-term and long-term to be preserved.
Obviously, BTS diluent prescription among the present invention is that every liter of diluent contains 37 gram glucose, 6 gram Trisodium Citrates, 0.75 gram Repone K, 1.25 gram sodium bicarbonates, 1.25 gram ethylene dinitrilotetra-acetic acid, 100000 unit penicillin and 100 milligrams of Streptomycin sulphates, cooling diluent prescription is: 100 milliliters of diluents contain lactose and 20 milliliters of yolk of 80 milliliter 11%, and the chill back liquid formula is: add 4% glycerine and 0.5% sodium lauryl sulfate in the cooling diluent.
Claims (10)
1. a Cryopreservation of Boar Semen pipe comprises and preserves pipe, it is characterized in that: preserve pipe and be double wall tube, two vessel ports of double wall tube are opposite.
2. Cryopreservation of Boar Semen pipe according to claim 1 is characterized in that: make with plastic material.
3. Cryopreservation of Boar Semen pipe according to claim 1 and 2 is characterized in that: double wall tube is a cylindrical tube.
4. Cryopreservation of Boar Semen pipe according to claim 3 is characterized in that: the volume of the annular container of double wall tube (2) is 5～15ml.
5. Cryopreservation of Boar Semen pipe according to claim 1 and 2 is characterized in that: the volume of the annular container of double wall tube (2) is 5～15ml.
6. Cryopreservation of Boar Semen method comprises:
A, semen collection and with one times of isothermal BTS (Beltsville thawing solution) diluted,
B, under 15 ℃ of conditions balance 2 hours, 1000 left the heart 10 minutes then,
Seminal plasma is removed in c, centrifugal back, and the sperm precipitation is with cooling off diluted, and making sperm concentration is 2 * 10
9Individual/milliliter.Seminal fluid after dilution balance 2 hours under 5 ℃ of conditions,
D and then with freeze-extender 1: 1 dilution, the final densities that makes sperm is 1 * 10
9It is individual/milliliter,
It is characterized in that: the seminal fluid after the chill back of above step process is packed in the annular container (2) of Cryopreservation of Boar Semen pipe of the present invention, then the Cryopreservation of Boar Semen pipe is inserted on the liquid nitrogen, stifling after several minutes, drop into directly that liquid nitrogen is medium-term and long-term to be preserved.
7. the method for Cryopreservation of Boar Semen according to claim 6, it is characterized in that: every Cryopreservation of Boar Semen pipe is adorned 5～15 milliliters of pig seminal fluid after the chill back.
8. according to the method for claim 6 or 7 described Cryopreservation of Boar Semen, it is characterized in that: the Cryopreservation of Boar Semen pipe that chill back pig seminal fluid will be housed is inserted on the liquid nitrogen surface 3 centimetres, and is stifling after 20 minutes, drops into directly that liquid nitrogen is medium-term and long-term to be preserved again.
9. the method for Cryopreservation of Boar Semen according to claim 8, it is characterized in that: BTS diluent prescription is that every liter of diluent contains 37 gram glucose, 6 gram Trisodium Citrates, 0.75 gram Repone K, 1.25 gram sodium bicarbonates, 1.25 gram ethylene dinitrilotetra-acetic acid, 100000 unit penicillin and 100 milligrams of Streptomycin sulphates, cooling diluent prescription is: 100 milliliters of diluents contain lactose and 20 milliliters of yolk of 80 milliliter 11%, and the chill back liquid formula is: add 4% glycerine and 0.5% sodium lauryl sulfate in the cooling diluent.
10. according to the method for claim 6 or 7 described Cryopreservation of Boar Semen, it is characterized in that: BTS diluent prescription is that every liter of diluent contains 37 gram glucose, 6 gram Trisodium Citrates, 0.75 gram Repone K, 1.25 gram sodium bicarbonates, 1.25 gram ethylene dinitrilotetra-acetic acid, 100000 unit penicillin and 100 milligrams of Streptomycin sulphates, cooling diluent prescription is: 100 milliliters of diluents contain lactose and 20 milliliters of yolk of 80 milliliter 11%, and the chill back liquid formula is: add 4% glycerine and 0.5% sodium lauryl sulfate in the cooling diluent.
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| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN2007101563988A CN101200688B (en) | 2007-10-30 | 2007-10-30 | Boar semen freezing preservation tube and method thereof |
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| CN2007101563988A CN101200688B (en) | 2007-10-30 | 2007-10-30 | Boar semen freezing preservation tube and method thereof |
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| CN101200688A true CN101200688A (en) | 2008-06-18 |
| CN101200688B CN101200688B (en) | 2013-03-27 |
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Cited By (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101671627B (en) * | 2008-09-12 | 2012-05-23 | 陈子江 | Vitrified refrigeration carrier and use method thereof |
| CN102657149A (en) * | 2012-05-08 | 2012-09-12 | 山西农业大学 | Ultra-high activity tubule semen freezing method for sheep |
| CN102986651A (en) * | 2012-12-24 | 2013-03-27 | 邓凯伟 | Method for freezing-storing, diluting and freezing boar semen particles |
| CN104322484A (en) * | 2014-10-10 | 2015-02-04 | 山东鑫基牧业有限公司 | Freezing and unfreezing method for boar semen |
| CN111903666A (en) * | 2020-09-03 | 2020-11-10 | 浙江省畜牧技术推广与种畜禽监测总站(浙江省农业机械试验鉴定推广总站) | Hu sheep semen freezing tube and thawing apparatus |
Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN2180515Y (en) * | 1993-12-22 | 1994-10-26 | 洪文虎 | Heating and refrigerating box for automobile |
| CN1596670A (en) * | 2004-07-27 | 2005-03-23 | 中国水产科学研究院黄海水产研究所 | Practicalization method for frozen preserving sperm of fish |
| WO2005032618A1 (en) * | 2003-09-30 | 2005-04-14 | Lifeforce Group Plc | Blood pack for leukocyte banking |
| CN1712519A (en) * | 2004-06-25 | 2005-12-28 | 华中农业大学 | Liquid for preserving pig semen |
-
2007
- 2007-10-30 CN CN2007101563988A patent/CN101200688B/en not_active Expired - Fee Related
Patent Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN2180515Y (en) * | 1993-12-22 | 1994-10-26 | 洪文虎 | Heating and refrigerating box for automobile |
| WO2005032618A1 (en) * | 2003-09-30 | 2005-04-14 | Lifeforce Group Plc | Blood pack for leukocyte banking |
| CN1712519A (en) * | 2004-06-25 | 2005-12-28 | 华中农业大学 | Liquid for preserving pig semen |
| CN1596670A (en) * | 2004-07-27 | 2005-03-23 | 中国水产科学研究院黄海水产研究所 | Practicalization method for frozen preserving sperm of fish |
Non-Patent Citations (2)
| Title |
|---|
| 李林林等: "猪精液冷冻技术研究进展", 《安徽农业科学》 * |
| 禚艳书等: "猪精液冷冻保存研究进展", 《江苏农业科学》 * |
Cited By (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101671627B (en) * | 2008-09-12 | 2012-05-23 | 陈子江 | Vitrified refrigeration carrier and use method thereof |
| CN102657149A (en) * | 2012-05-08 | 2012-09-12 | 山西农业大学 | Ultra-high activity tubule semen freezing method for sheep |
| CN102657149B (en) * | 2012-05-08 | 2013-09-18 | 山西农业大学 | Ultra-high activity tubule semen freezing method for sheep |
| CN102986651A (en) * | 2012-12-24 | 2013-03-27 | 邓凯伟 | Method for freezing-storing, diluting and freezing boar semen particles |
| CN104322484A (en) * | 2014-10-10 | 2015-02-04 | 山东鑫基牧业有限公司 | Freezing and unfreezing method for boar semen |
| CN111903666A (en) * | 2020-09-03 | 2020-11-10 | 浙江省畜牧技术推广与种畜禽监测总站(浙江省农业机械试验鉴定推广总站) | Hu sheep semen freezing tube and thawing apparatus |
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| CN101200688B (en) | 2013-03-27 |
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