CN104542572A - Method for manufacturing straw frozen semen of gayal and application - Google Patents
Method for manufacturing straw frozen semen of gayal and application Download PDFInfo
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- CN104542572A CN104542572A CN201310496109.4A CN201310496109A CN104542572A CN 104542572 A CN104542572 A CN 104542572A CN 201310496109 A CN201310496109 A CN 201310496109A CN 104542572 A CN104542572 A CN 104542572A
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Abstract
The invention discloses a method for manufacturing the straw frozen semen of a gayal and application of the method. The gayal is still a semi-wild and semi-domestic animal, is a second-grade wild animal under state protection, and is only native to the area of Dulongjiang of the Derung Nationality and Nu Nationality Autonomous County of Gongshan of Yunnan in China. The straw frozen semen of the gayal (the frozen semen for short) is manufactured by sealing the collected fresh semen of the gayal in a miniature plastic straw of 0.25 ml after the fresh semen of the gayal is processed, freezing the semen according to a specific program and a specific freezing speed by utilizing liquid nitrogen as a cold source, and storing the semen in an ultra-low-temperature environment in a frozen form, so that the aim of storing the semen of the gayal for a long time is achieved. The manufacturing of the straw frozen semen of the gayal is favorable for long-term storage of the germ plasm of the gayal, and prevents the germ plasm depression and the gene loss of the gayal species.
Description
Technical field:
The Straw Frozen Semen that the present invention relates to ox makes.
Background technology:
Straw Frozen Semen refers to seminal fluid splendid attire after treatment in plastic straw, is immersed in the medium-term and long-term preservation of liquid nitrogen after ultralow temperature snap frozen, and the frozen semen that can take at any time.
The preparation of stock bull before Bian essence: stock bull macroscopic features should meet the national standard of kind, Dairy bull general 16 monthly ages start pilot production essence, and beef breeding bull general 18 monthly ages start pilot production essence.Breed frequently in situation stock bull; feed of animal origin should be increased; because protein is conducive to the formation of seminal fluid and sperm. according to feeding and management code, stock bull also will be carried out supplementary feeding, herds, moves, expelling parasite, epidemic prevention, the routine work such as foot paring and semen collection training.
Summary of the invention:
Gayal is a kind ofly still in the domestic domestic animals of semi-wild half, is national wildlife under second class protection, only originates in Drung-Nu Autonomous County of Gongshan, Yunnan Province Dulong River one band in China.Gayal Straw Frozen Semen (hereinafter referred to as freezing essence) refers to the gayal fresh semen that will collect, after treatment, seminal fluid is encapsulated in miniature (0.25ml) plastic straw, utilize liquid nitrogen for low-temperature receiver, undertaken freezing by specific program and chilling rate, under making seminal fluid be stored in ultra-low temperature surroundings with the form freezed, reach long-term object of preserving gayal seminal fluid.Gayal straw frozen semen be manufactured with the long-term preservation being beneficial to its genetic material, prevent gayal population kind matter fail and gene loss.
Embodiment:
The preparation method of gayal straw frozen semen is:
1. the selection of gayal bull: the bull for making straw frozen semen will select build large, and the obvious bull of gayal feature carries out domestication and instruction.
2. semen collection instruction: application method of luring carries out semen collection instruction, by sex inducement to the Progressive symmetric erythrokeratodermia stimulation repeatedly of gayal bull, make the generation reflection of bull, expression power behavior, carry out artificial vagina semen collection training, when bull heat strong also mounting dummy teaser (or companion), in time close to bull, stroke bull buttocks, external genital and foreskin, when bull heat is abundant with ready artificial vagina semen collection.
3. semen collection method: Pseudopyloric metaplasia, during with artificial vagina semen collection, artificial vagina inner tube of a tyre temperature is 38 ~ 42 DEG C, suitably regulates artificial vagina pressure and lubricity.During semen collection, semen collection person will note person poultry safety, unruffled, bold but cautious, skilled operation, nature first, stand on the right side of bull, when bull mounting, catching the best opportunity takes advantage of a situation semen collection, prevents from causing the unhappy of bull during semen collection and affects normal semen collection and form pernicious habit.
4. the preparation of gayal freeze-extender: reagent needed for precise, mixes, dissolves, filters and sterilizes and be stand-by.
5. straw frozen semen making apparatus: phase contrast microscope, spectrophotometer, MRS-3 automatic packer, program-controlled freezing instrument, liquid nitrogen biological container, liquid nitrogen, mini-straw, thumb pipe, refrigerator, low-temperature operation cabinet, freezing semen frame, slide, graduated cylinder, measuring cup, centrifuge tube, thermostat water bath, pipettor and dilution etc.
6. the process of gayal seminal fluid; First the gayal seminal fluid collected measures volume of semen with graduated centrifuge tube or by weight method; Measure sperm concentration: get 0.7%NaCl solution 3960 microlitre in cuvette, then get former smart 40 microlitres of gayal and add NaCl solution and make both fully mixings, put into spectrophotometric colo groove and measure sperm concentration; Use phase-contrast microscopy Fresh semen activity.
7. the dilution of gayal seminal fluid: Fresh semen activity more than 60%, the gayal seminal fluid freeze-extender of sperm concentration 2.0 hundred million/more than ml dilutes, and every dosage sperm adds up to 015 ~ 0.20 hundred million, effective number of sperm 450 ~ 8,000,000.The packing of seminal fluid: the gayal seminal fluid MRS-3 automatic packer after dilution is at room temperature divided in mini-straw, sealing and lettering.The balance of gayal straw semen: the straw semen after packing is positioned over refrigerator and balances 2 ~ 4 hours at the temperature of+4 DEG C.
8. gayal straw semen is freezing: the gayal seminal fluid after balance is neatly placed on freezing semen frame at about+4 DEG C temperature, then freezing semen frame and seminal fluid are put into freezing 6 ~ 10 minutes of the liquid nitrogen vapor of-130 ~-140 DEG C, then the straw frozen semen after freezing is immersed and carry out ultralow temperature in liquid nitrogen and freeze.Or it is freezing with program-controlled freezing instrument, freezing curve is :+4 DEG C to-10 DEG C, chilling rate is 5 DEG C/mn.-10 DEG C to-100 DEG C, chilling rate is 40 DEG C/mn.-100 is DEG C to-140 DEG C, chilling rate is 20 DEG C/mn, then the straw frozen semen after freezing is immersed and carries out ultralow temperature in liquid nitrogen and freeze.
9. thawing of gayal straw frozen semen checks and preserves: the gayal straw frozen semen after freezing randomly draws 1 ~ 2, the water-bath of putting into 40 ± 2 DEG C is immediately thawed 5 ~ 20 seconds, inspection is thawed vigor, the vigor of thawing can reach 35 ~ 45%, and the gayal straw frozen semen packaging of the vigor more than 30% that thaws puts into the medium-term and long-term preservation of liquid nitrogen.
10. the use of gayal straw frozen semen: namely the gayal straw frozen semen that after freeze-thaw, vigor reaches more than 30% can be used for artificial insemination, the effective number of sperm of the every dosage of suitable reduction, do not affect gayal straw frozen semen semen deposition conception rate, non-return rate can reach more than 50%.
Claims (10)
1. gayal selects the bull that build is large, it is characterized in that obvious bull carries out domestication and instruction.
2. semen collection instruction, it is characterized in that application method of luring carries out semen collection instruction, by sex inducement to the Progressive symmetric erythrokeratodermia stimulation repeatedly of gayal bull, make the generation reflection of bull, expression power behavior, carry out artificial vagina semen collection training, when bull heat strong also mounting dummy teaser (or companion), in time close to bull, stroke bull buttocks, external genital and foreskin, when bull heat is abundant with ready artificial vagina semen collection.
3. semen collection method, it is characterized in that Pseudopyloric metaplasia, during with artificial vagina semen collection, artificial vagina inner tube of a tyre temperature is 38 ~ 42 DEG C, and suitably regulate artificial vagina pressure and lubricity, during semen collection, semen collection person will note person poultry safety, unruffled, bold but cautious, skilled operation, nature first, stand on the right side of bull, when bull mounting, catching the best opportunity takes advantage of a situation semen collection, prevents from causing the unhappy of bull during semen collection and affects normal semen collection and form pernicious habit.
4. the preparation of gayal freeze-extender, is characterized in that reagent needed for precise, mixes, dissolves, filters and sterilize stand-by.
5. straw frozen semen making apparatus, is characterized in that phase contrast microscope, spectrophotometer, MRS-3 automatic packer, program-controlled freezing instrument, liquid nitrogen biological container, liquid nitrogen, mini-straw, thumb pipe, refrigerator, low-temperature operation cabinet, freezing semen frame, slide, graduated cylinder, measuring cup, centrifuge tube, thermostat water bath, pipettor and dilution etc.
6. the process of gayal seminal fluid, it is characterized in that first the gayal seminal fluid collected measures volume of semen with graduated centrifuge tube or by weight method, measure sperm concentration: get 0.7%NaCl solution 3960 microlitre in cuvette, then get former smart 40 microlitres of gayal to add NaCl solution and make both fully mixings, put into spectrophotometric colo groove and measure sperm concentration, use phase-contrast microscopy Fresh semen activity.
7. the dilution of gayal seminal fluid, it is characterized in that Fresh semen activity more than 60%, the gayal seminal fluid freeze-extender of sperm concentration 2.0 hundred million/more than ml dilutes, every dosage sperm adds up to 015 ~ 0.20 hundred million, effective number of sperm 450 ~ 8,000,000, the packing of seminal fluid: the gayal seminal fluid MRS-3 automatic packer after dilution is at room temperature divided in mini-straw, sealing and lettering, the balance of gayal straw semen: the straw semen after packing is positioned over refrigerator and balances 2 ~ 4 hours at the temperature of+4 DEG C.
8. gayal straw semen is freezing, it is characterized in that the gayal seminal fluid after balancing neatly is placed on freezing semen frame at about+4 DEG C temperature, then freezing semen frame and seminal fluid are put into freezing 6 ~ 10 minutes of the liquid nitrogen vapor of-130 ~-140 DEG C, then the straw frozen semen after freezing is immersed and carry out ultralow temperature in liquid nitrogen and freeze.Or it is freezing with program-controlled freezing instrument, freezing curve is :+4 DEG C to-10 DEG C, chilling rate is 5 DEG C/mn.-10 DEG C to-100 DEG C, chilling rate is 40 DEG C/mn.-100 is DEG C to-140 DEG C, chilling rate is 20 DEG C/mn, then the straw frozen semen after freezing is immersed and carries out ultralow temperature in liquid nitrogen and freeze.
9. thawing of gayal straw frozen semen checks and preserves, it is characterized in that freezing after gayal straw frozen semen randomly draw 1 ~ 2, the water-bath of putting into 40 ± 2 DEG C is immediately thawed 5 ~ 20 seconds, inspection is thawed vigor, the vigor of thawing can reach 35 ~ 45%, and the gayal straw frozen semen packaging of the vigor more than 30% that thaws puts into the medium-term and long-term preservation of liquid nitrogen.
10. the use of gayal straw frozen semen, namely the gayal straw frozen semen that after it is characterized in that freeze-thaw, vigor reaches more than 30% can be used for artificial insemination, the effective number of sperm of the every dosage of suitable reduction, do not affect gayal straw frozen semen semen deposition conception rate, non-return rate can reach more than 50%.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
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| CN201310496109.4A CN104542572A (en) | 2013-10-22 | 2013-10-22 | Method for manufacturing straw frozen semen of gayal and application |
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| CN201310496109.4A CN104542572A (en) | 2013-10-22 | 2013-10-22 | Method for manufacturing straw frozen semen of gayal and application |
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN106106433A (en) * | 2016-06-22 | 2016-11-16 | 湖北省畜禽育种中心 | A kind of Binglangjiang waterbuffalo freezing of semen process |
| CN111345283A (en) * | 2020-03-31 | 2020-06-30 | 上海市农业科学院 | Method for freezing and storing buffalo semen |
-
2013
- 2013-10-22 CN CN201310496109.4A patent/CN104542572A/en active Pending
Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN106106433A (en) * | 2016-06-22 | 2016-11-16 | 湖北省畜禽育种中心 | A kind of Binglangjiang waterbuffalo freezing of semen process |
| CN106106433B (en) * | 2016-06-22 | 2019-04-16 | 湖北省畜禽育种中心 | A kind of Binglangjiang waterbuffalo freezing of semen process |
| CN111345283A (en) * | 2020-03-31 | 2020-06-30 | 上海市农业科学院 | Method for freezing and storing buffalo semen |
| CN111345283B (en) * | 2020-03-31 | 2022-02-15 | 上海市农业科学院 | Method for freezing and storing buffalo semen |
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Application publication date: 20150429 |