CN1299636A - New method of producing frozen bull semen - Google Patents
New method of producing frozen bull semen Download PDFInfo
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- CN1299636A CN1299636A CN 00110459 CN00110459A CN1299636A CN 1299636 A CN1299636 A CN 1299636A CN 00110459 CN00110459 CN 00110459 CN 00110459 A CN00110459 A CN 00110459A CN 1299636 A CN1299636 A CN 1299636A
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Abstract
The productin method of ox or other animal frozen semen includes the following steps: packaging ox tubule semen in proper packet under the condition of before freezing and placing it in freezing basket, pressing all the packets close together or binding them to mare the ox semen tubules vertically and densely arrange, then progressively coaking them into liquid nitrogen, and controlling soaking speed to control the cooling rate of semen.
Description
The invention belongs to a kind of production method of product, particularly the straw semen of cattle vertically dense arrangement be immersed in a kind of new method of making the cow frozen semen product among the liquid nitrogen vertically with certain speed.The invention belongs to Animal Science animal reproduction technical field.
The frozen semen of cattle is born in nineteen fifty.A kind of product that is meant a seminal fluid of the bull that collects to freeze in ultra-low temperature surroundings after suitably handling and forms can be made long preservation to improve the utilization ratio of outstanding bull, accelerates the reproduction speed of breeding.Be widely used in the animal husbandry at present.
(1) dosage form of cow frozen semen
The cow frozen semen product has two kinds of dosage forms at present, i.e. granular pattern and tubule type.Granular pattern is the seminal fluid after handling to be dripped to freeze holding to freeze make hemispheric product on the face, because the shortcoming that there is easy pollution and be difficult for sign is only used in a few countries such as China now.Tubule type frozen semen is that the seminal fluid after the processing is sub-packed in the product of lowering the temperature and freezing to make behind the north material of the special-purpose polrvinyl chloride tubule with Special Equipment.Be divided into two kinds of 0.25ml and 0.5ml again according to the difference of capacity, length all is 133mm, and each is 2mm and 3mm approximately for an external diameter.
(2) key factor of the technological process of production of cattle tubule type frozen semen product and decision product quality
At present, the production stage of cattle tubule type frozen semen is followed successively by semen collection, semen quality inspection, dilution, cooling, balance, packing, freezing (freezing), packing, warehouse-in.Wherein freezing this link is the key factor of decision product quality.The early stage refrigerant of using is dry ice (solid carbon dioxide).What use now is liquid nitrogen, and its boiling point be-195.8C, makes after airborne nitrogen is utilized Special Equipment to compress to lower the temperature.When being refrigerant with the liquid nitrogen, freezing rate (is meant 0-5 ℃ refrigerating process the unit interval decrease of temperature value of seminal fluid before freezing, also claim rate of temperature fall) the too fast or slow excessively quality of freezing the back product that all can influence, when freezing rate is in a certain scope, freeze the best in quality of essence.
(3) the existing method of cattle tubule frozen semen production both at home and abroad
The method of existing cattle tubule frozen semen production is called as liquid nitrogen steam fumigation and steaming method.The state of liquid nitrogen steam is divided into two kinds of immobilized liquid nitrogen steam fumigation and steaming method and mobile liquid nitrogen steam fumigation and steaming method again when freezing.The stifling ratio juris of immobilized liquid nitrogen steam is the temperature difference of utilizing between straw semen and its bearing system (metal rack, frame, net etc.) and liquid nitrogen and liquid nitrogen steam, liquid nitrogen vaporization is absorbed heat and seminal fluid is lowered the temperature to freeze.The arrangement mode of tubule can be divided into two types to fumigation and steaming method when freezing again.A kind of is horizontal formula, and another kind is vertical arranged type.Which kind of arrangement mode no matter, prior art all requires must keep suitable distance between tubule, carries out dispersed arrangement.The method of keeping at a distance is that tubule flatly is arranged in parallel among jagged freezing grid the square hole last or separate mesh of the circular freezing basket of insertion vertically.
Adopting immobilized liquid nitrogen steam fumigation and steaming method to carry out frozen semen production carries out in low-temperature (low temperature) vessel.Fill the formation temperature gradient of low-temperature (low temperature) vessel meeting nature between liquid nitrogen surface and vessel port plane of liquid nitrogen, near more apart from liquid nitrogen surface, temperature is just low more, and is just big more with the temperature difference of freezing preceding tubule.The basic operational steps of this method is: at first add a certain amount of liquid nitrogen in low-temperature (low temperature) vessel, the seminal fluid tubule flatly is parallel on jagged freezing, the tubule spacing is about 6mm, flatly put into the top of low-temperature (low temperature) vessel liquid nitrogen surface then in the lump, making lowers the temperature in the immobilized liquid nitrogen steam of seminal fluid tubule on liquid nitrogen surface freezes.This method is rough to the adjusting of rate of temperature fall, only relies on the height distance of freezing and tubule plane separation liquid nitrogen surface to control rate of temperature fall.
The stifling ratio juris of mobile liquid nitrogen steam is on the basis that static liquid nitrogen steam conducts heat, and accelerates the speed of heat exchange, makes speeding up of lowering the temperature and freeze.The realization of this method needs the cooperation of accurate machinery, electronic installation or computer controlled automatic device, so that accurately control rate of temperature fall.The producing method of flowing steam has following four kinds: spray into liquid nitrogen steam from the refrigerating chamber top with certain speed 1..2. refrigerating chamber bottom dress fan drives the liquid nitrogen vapor recycle.3. metal grill heating liquid nitrogen produces mobile liquid nitrogen steam.4. in liquid nitrogen, charge into the room temperature compressed nitrogen and obtain mobile liquid nitrogen steam.The common feature of these methods is the freezing rates by how many next indirect control straw semens of control liquid nitrogen steam generation.
The objective of the invention is to raise labour productivity: the quality that improves cattle tubule frozen semen product by improving every batch of refrigerated quantity; Improve the quantity of cattle tubule frozen semen product, thereby reduce the number of animals raised of breeding oxen; Simplify the structure and the technological process of production of freezing equipment.
Feature of the present invention is, carrying out the freezing clear liquid of cattle tubule when producing, and seminal fluid tubule dense arrangement vertically, is immersed in certain decrease speed vertically and lowers the temperature among the liquid nitrogen and freeze.
The present invention packs the cattle straw semen in the packing material of suitable size into (gauze bag, the little tube of plastics etc.) and makes each packing jam-packed or they are bundled this dual mode in freezing basket to make the intensive vertical arrangement of seminal fluid tubule, and at normal temperatures straw semen is packed before cooling is freezed.Existing method flatly goes up straw semen or lower the temperature then vertically and freeze at a distance of suitably being arranged in jagged freezing abreast earlier among the grid square hole of the separate mesh that suitably inserts circular freezing basket, freeze the back and pack under ultralow temperature (195.8 ℃) environment.
The present invention relies on little by little to be immersed liquid nitrogen to freezing basket and seminal fluid tubule wherein and lowers the temperature and freeze, and relies on the speed of freezing basket immersion (decline) accurately to control the rate of temperature fall of seminal fluid.The average speed that immerses liquid nitrogen decide on the quantity of each refrigerated tubule, quantity after a little while, average decline (immersion) speed can be omited soon; Quantity for a long time, average (immersion) speed that descends should be omited slowly.Be generally average per minute decline 5mm～15mm, and when beginning (immersion) speed that descends slower, subsequently should be more and more faster.Average about 10～25 minutes by the complete submergence of liquid nitrogen.
The method that the present invention adopts is called intensive immersion method, considers from the angle of Cryo Conduction, can be described as intensive radiant heat method again.The principle of the freezing cattle straw semen of immersion method is dexterously the conduction of heat of the convection heat transfer' heat-transfer by convection of horizontal direction and vertical direction to be combined, and relies on submerged speed accurately to control the rate of temperature fall of seminal fluid in the tubule.
The ultimate principle one of immersion method is to have utilized the vessel port plane of liquid nitrogen container and liquid nitrogen inside to exist the temperature difference about 200 ℃, along with tubule apart from the height of liquid nitrogen surface be dipped among the liquid nitrogen, can make that tubule (even in batches maximum) is inner to obtain about 1 ℃/minute～200 ℃/minute cooling rate, this scope has comprised the best cooling rate or the best cooling rate scope of freezing of semen fully.The 2nd,, adopt immersion method to carry out the cattle straw semen when freezing, the immersion of tubule (decline) the speed cooling rate inner with it exists a kind of fixed corresponding relation, thereby the accurate control of mechanical displacement has been reached accurate control to cooling rate with regard to simple ground connection.The 3rd,, as long as control is proper, can accomplish that the tubule inner rigid reaches-130 ℃ and just immerses liquid nitrogen, reach-130 ℃ by best cooling rate before this.The 4th,, lower the temperature by convection heat transfer' heat-transfer by convection with existing fumigation and steaming method and to compare, immersion method mainly is based on the conduction of heat cooling, because the material polrvinyl chloride of tubule and the solution of Qi Nei are the materials of heat conductivity extreme difference, this each cross section of tubule of just having guaranteed vertical state did not reach dangerous warm area in its inside as yet begin cooling with best cooling rate before.
(1) increases substantially the volume of production of every batch of cow frozen semen product, thereby improved labor productivity.
The present invention is the liquid nitrogen container of 260mm if adopt diameter, each maximum output is 5300 0.5ml straw semens (being equivalent to 13250 0.25ml straw semens), the cross-sectional unit area of pressing the low-temperature (low temperature) vessel that is adopted calculates, and the present invention has improved 8～83 times than prior art.Same volume of production adopts the present invention to make the labour force of labor time and employing reduce to 1 hour * 1 people from 5 hours * 4 people.
(2) improved the quality of this product of cow frozen semen.
A most important index weighing cooling freezing technology level height is the height that freezes back sperm anabiosis rate.The sperm anabiosis rate is meant that the motility rate of the back sperm that thaws accounts for the percentage ratio that freezes preceding sperm motility rate after freezing.China's existing level is 50%～60%, can reach 70%～85% behind employing the present invention.
(3) improved every total quantity of bull year producing frozen semen.
Because the quality of this product of frozen semen is just determining the sperm sum that each dosage form should comprise freezing before, along with the raising of frozen semen quality, the sperm that every tubule should comprise before freezing is total just can be reduced.Just can increase the quantity of product or the number of animals raised of minimizing breeding oxen according to the market situation decision.
(4) simplified the technological process that cow frozen semen is produced.
Existing procedure is the semen collection → semen quality inspection → dilution → cooling first time → second time dilution → balance → packing → dress basket (shelving) → freezing → inspect by random samples → be packaged into bag → warehouse-in.The present invention adopts the refrigerated method in packing back earlier, makes above-mentioned process simplification be: packing → be packaged into bag → frame up → freezing → sampling observation → warehouse-in.Production procedure after the simplification has improved labor efficiency, avoided being packaged into the occupation disease that bag work institute may cause around the low temperature environment for a long time, avoided in being packaged into bag process because of looking into the decline of the product quality that the seminal fluid intensification is caused in the tubule that the temporary transient disengaging of number liquid nitrogen causes.
(5) be packaged as basic frozen units with one.
Compare for basic frozen units with one with prior art, make that the seminal fluid of bull of Different Individual, different cultivars can be in same freezing basket, in same batch, carry out freezing and be unlikely to obscure.
(6) saved the use amount of liquid nitrogen.
When adopting the present invention to produce 5300 0.5ml tubule clear liquids, the amount that consumes liquid nitrogen is about 8 liters, and compared with prior art, consumption is minimum.
(7) device structure that the present invention relied on is simple, cost is low, and the present invention is realized easily.
Realize that special equipment of the present invention describes in detail in the utility application file of by name " the program control frigorimeter of intensive immersion seminal fluid " that I propose on the same day.
It is multiple to realize that mode of the present invention has.Every energy by certain mode control a cantilever and (or) devices such as the machinery of the decline of metal bracelet etc., electronics, hydraulic pressure all can realize the present invention.The applicant thinks that realization best way of the present invention is the mechanical lifting gear of accurately being controlled by computer program, and this equipment is described in detail in the utility application file of " the program control frigorimeter of intensive immersion seminal fluid " by name that I propose on the same day.Below, with the accompanying drawing of this embodiment to realizing that the present invention is illustrated more clearly in.
Realize that operating procedure of the present invention is:
1. pack.After the seminal fluid packing finishes, go out 50 or 100 straw semens with code-disc device sign indicating number, they are concentrated into a bundle (bundle) with handgrip, pack into to freeze smart production in the special-purpose gauze bag (or the little tube of plastics).The tampon end of tubule is towards the bottom of gauze bag.Less than 50 or 100 s', pack into by actual quantity.Install the cord of back tension bag mouth and prick upper bag mouth.
2. frame up.Two kinds of methods are arranged.(1) (diameter is that the heap(ed) capacity of the freezing basket 3 of 255mm is: the 0.5ml tubule is 5300 when packing maximum into, 0.25ml tubule is 13250), earlier freezing basket 3 is kept flat, the front is towards the operator, 4 one bags of one bag of ground of tubule bag are vertically piled up in freezing basket, mutual jam-packed up between each bag is till freezing basket 3 no longer can hold.The tying of tubule bag 4 outwardly.When (2) being less than the tubule bag 4 of maximum when packing into, earlier 4 one bags of one bag of ground of tubule bag are piled up, apart suitably bind with two fine rule ropes then, the column 7 from the slit between the bag of middle along freezing basket 3 stands on this a bundle straw semen bag among freezing basket 3 vertically.Rely on this dual mode of pack and jam-packed (encircling) dress basket to realize the dense arrangement of tubule.
3. injection liquid nitrogen.Add an amount of liquid nitrogen 2 in the liquid nitrogen container 1 of frigorimeter, liquid nitrogen surface is dark apart from vessel port plane 165～185mm.Cover lucite cover plate 5, made the liquid nitrogen container pre-cooling 10-15 minute.
4. rise the cantilever 11 of frigorimeter.Activate " the fast liter " button 12 of controlling on the software interface with mouse or keyboard.
5. input data.Measure liquid nitrogen surface apart from the planar distance of vessel port (mm), measure seminal fluid in the tubule freeze preceding temperature (℃), be input in the computer together with these 3 data of tubule number (propping up).
6. hang freezing basket.Wait that being equipped with the freezing basket 3 that freezes straw semen is suspended on the hook 9 of metallic bond 10 lower ends that link to each other with frigorimeter cantilever 11.Hook 9 is drilled with circular hole 6 from the center poly (methyl methacrylate) plate 5 passes, and penetrates in the vertical circular hole 8 of column 7 of freezing basket 3.
7. carry out freezing.Activate " freezing " button 13.Freezing basket will descend automatically by pre-set programs.
8. behind the program end of run, freezing basket 3 has dropped to the bottom of container, removes hook 9, take poly (methyl methacrylate) plate 5 away, cantilever 11 is rotated on one side, with the long handle tweezers tubule bag 4 that freezes to finish is transferred in the interior liquid nitrogen of storage type liquid nitrogen container, sampling observation back warehouse-in is preserved.
Claims (6)
1. produce the particularly method of the tubule type frozen semen product of cattle of frozen semen product for one kind, its feature one is the intensive vertical arrangement of straw semen, makes the spacing between tubule be compressed to minimum degree; The 2nd, make straw semen be immersed in gradually to lower the temperature among the liquid nitrogen with certain speed vertically and freeze.
2. according to the method for claim 1, feature of the present invention is the cattle straw semen to be packed in the packing material of suitable size by (gauze bag, the little tube of plastics etc.) and made each packing jam-packed or they are bundled this dual mode in freezing basket make the intensive vertical arrangement of seminal fluid tubule.
3. according to the method for claim 1 or 2, feature of the present invention is before cooling is freezed straw semen to be packed.
4. according to the method for claim 1, feature of the present invention be the average speed that immerses liquid nitrogen be straw semen together with freezing basket per minute decline 5mm～15mm, and (immersion) speed that descends during beginning is slower, subsequently should be more and more faster.Average about 10～25 minutes by the complete submergence of liquid nitrogen.
5. according to the method for claim 1 or 4, it is fixing that feature of the present invention is that the quantity of the straw semen at every turn produced does not require, quantity after a little while, average (immersion) speed that descends can be omited soon; Quantity for a long time, average (immersion) speed that descends should be omited slowly.
6. according to the method for claim 1, the present invention also is applicable to the frozen semen production of other animal.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CNB001104594A CN1150862C (en) | 2000-05-29 | 2000-05-29 | New method of producing frozen bull semen |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CNB001104594A CN1150862C (en) | 2000-05-29 | 2000-05-29 | New method of producing frozen bull semen |
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| Publication Number | Publication Date |
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| CN1299636A true CN1299636A (en) | 2001-06-20 |
| CN1150862C CN1150862C (en) | 2004-05-26 |
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| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CNB001104594A Expired - Fee Related CN1150862C (en) | 2000-05-29 | 2000-05-29 | New method of producing frozen bull semen |
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Cited By (10)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1331390C (en) * | 2003-12-24 | 2007-08-15 | 甘孜州家畜改良站 | Method for making frozen spermatozoon of yak |
| CN100383237C (en) * | 2004-03-12 | 2008-04-23 | 中国科学院昆明动物研究所 | Low-temp freezing method for storing sperm of gayal |
| CN102125026A (en) * | 2011-01-26 | 2011-07-20 | 沈阳航天新光低温容器制造有限责任公司 | Program-controlled freezing instrument |
| CN102616472A (en) * | 2012-03-07 | 2012-08-01 | 上海交通大学 | Integrated device for fumigating, freezing, storing and transporting gametes based on liquid nitrogen |
| CN103750919A (en) * | 2014-01-20 | 2014-04-30 | 北京天新福医疗器材有限公司 | Method for freezing and forming artificial nerve sheath |
| CN104488851A (en) * | 2014-12-11 | 2015-04-08 | 燕海峰 | Full-automatic multi-step freezing instrument |
| CN106342789A (en) * | 2016-10-13 | 2017-01-25 | 深圳韦拓生物科技有限公司 | Cell vitrification freezing treatment equipment and method |
| CN111110393A (en) * | 2019-12-09 | 2020-05-08 | 中南百草原集团有限公司 | Bovine semen extraction method |
| US10660327B2 (en) | 2015-07-02 | 2020-05-26 | Xiaojun Xing | Multilayer insulation of biological samples and cooling by immersing in a cryogenic liquid for cryopreservation |
| CN111990394A (en) * | 2020-07-27 | 2020-11-27 | 安徽农业大学 | Method for filling frozen semen tubules into finger-shaped tubes |
Families Citing this family (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1295956C (en) * | 2004-07-27 | 2007-01-24 | 中国水产科学研究院黄海水产研究所 | Practicalization method for frozen preserving sperm of fish |
-
2000
- 2000-05-29 CN CNB001104594A patent/CN1150862C/en not_active Expired - Fee Related
Cited By (12)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1331390C (en) * | 2003-12-24 | 2007-08-15 | 甘孜州家畜改良站 | Method for making frozen spermatozoon of yak |
| CN100383237C (en) * | 2004-03-12 | 2008-04-23 | 中国科学院昆明动物研究所 | Low-temp freezing method for storing sperm of gayal |
| CN102125026A (en) * | 2011-01-26 | 2011-07-20 | 沈阳航天新光低温容器制造有限责任公司 | Program-controlled freezing instrument |
| CN102616472A (en) * | 2012-03-07 | 2012-08-01 | 上海交通大学 | Integrated device for fumigating, freezing, storing and transporting gametes based on liquid nitrogen |
| CN103750919A (en) * | 2014-01-20 | 2014-04-30 | 北京天新福医疗器材有限公司 | Method for freezing and forming artificial nerve sheath |
| CN104488851A (en) * | 2014-12-11 | 2015-04-08 | 燕海峰 | Full-automatic multi-step freezing instrument |
| US10660327B2 (en) | 2015-07-02 | 2020-05-26 | Xiaojun Xing | Multilayer insulation of biological samples and cooling by immersing in a cryogenic liquid for cryopreservation |
| CN106342789A (en) * | 2016-10-13 | 2017-01-25 | 深圳韦拓生物科技有限公司 | Cell vitrification freezing treatment equipment and method |
| CN106342789B (en) * | 2016-10-13 | 2022-07-15 | 深圳韦拓生物科技有限公司 | Cell vitrification freezing treatment equipment and treatment method thereof |
| CN111110393A (en) * | 2019-12-09 | 2020-05-08 | 中南百草原集团有限公司 | Bovine semen extraction method |
| CN111110393B (en) * | 2019-12-09 | 2022-08-09 | 中南百草原集团有限公司 | Bovine semen extraction method |
| CN111990394A (en) * | 2020-07-27 | 2020-11-27 | 安徽农业大学 | Method for filling frozen semen tubules into finger-shaped tubes |
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| Publication number | Publication date |
|---|---|
| CN1150862C (en) | 2004-05-26 |
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