CN106070184A - The freezing of a kind of pig semen and defreezing method - Google Patents
The freezing of a kind of pig semen and defreezing method Download PDFInfo
- Publication number
- CN106070184A CN106070184A CN201610374193.6A CN201610374193A CN106070184A CN 106070184 A CN106070184 A CN 106070184A CN 201610374193 A CN201610374193 A CN 201610374193A CN 106070184 A CN106070184 A CN 106070184A
- Authority
- CN
- China
- Prior art keywords
- freezing
- seminal fluid
- semen
- pig
- pig semen
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Pending
Links
Classifications
-
- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01N—PRESERVATION OF BODIES OF HUMANS OR ANIMALS OR PLANTS OR PARTS THEREOF; BIOCIDES, e.g. AS DISINFECTANTS, AS PESTICIDES OR AS HERBICIDES; PEST REPELLANTS OR ATTRACTANTS; PLANT GROWTH REGULATORS
- A01N1/00—Preservation of bodies of humans or animals, or parts thereof
- A01N1/02—Preservation of living parts
- A01N1/0278—Physical preservation processes
- A01N1/0284—Temperature processes, i.e. using a designated change in temperature over time
Landscapes
- Life Sciences & Earth Sciences (AREA)
- Health & Medical Sciences (AREA)
- Engineering & Computer Science (AREA)
- Dentistry (AREA)
- General Health & Medical Sciences (AREA)
- Wood Science & Technology (AREA)
- Zoology (AREA)
- Environmental Sciences (AREA)
- Agricultural Chemicals And Associated Chemicals (AREA)
Abstract
The invention discloses freezing and the defreezing method of a kind of pig semen, freezing method including pig semen, the defreezing method of pig semen and the detection of pig semen, the freezing method of pig semen comprises the following steps: gathers breeding boar seminal fluid and is measured its density, dilution.Seminal fluid after dilution is stood, balance of lowering the temperature afterwards.Seminal fluid after cooling is centrifugal treating at a temperature of 14 15 DEG C.The diluent I being sequentially added into, diluent II carry out subpackage.The seminal fluid that subpackage is good proceeds by freezing, and liquid nitrogen freezing preserves.Defreezing method is as follows: taking rapidly the pig semen of freezing, temperature rises to 5 DEG C with the speed of 50 DEG C/min from 140 DEG C, then rises to 5 DEG C of defrostings with the speed of 3 DEG C/min from 5 DEG C.The present invention be penetration and promotion pig freeze essence produce on application provide technology.
Description
Technical field
The invention belongs to the freezen protective technical field of pig semen, specifically, relate to the freezing reconciliation of a kind of pig semen
Freeze method.
Background technology
Artificial insemination (artificial insemination, the AI) technology of pig, i.e. utilizes manual method to take boar
Seminal fluid, carries out quality inspection, dilution preservation etc. to it and processes, then be transported to oestrus in Sow Genital Organs by seminal fluid with apparatus,
Make it become pregnant.Artificial insemination can improve the breeding efficiency of boar, and compared with this friendship, during artificial insemination, one outstanding breeding boar is annual
The sow of breeding can improve 10～20 times.Due to the raising of breeding boar utilization rate, accelerate breed improvement, Swine Production can be given
Breeding with pig brings huge benefit；Reduce the number of animals raised of boar, save feeding and management expense；Natural mating can be overcome
Difficult point, the difficulties such as too big in male and female pig physique difference and some reproductive tract are abnormal；Outstanding boar can be expanded in the time and geographically
Utilization rate, provide conveniently for the exchange of outstanding kind of pig semen；Additionally, artificial insemination can also effectively prevent male and female pig many
Plant epidemic disease possibility contagious.Just because of artificial insemination, there is in Swine Production lot of superiority so that this technology
Widelyd popularize in many developed countries of raising pigs.But, seminal fluid used in the artificial insemination of pig both at home and abroad mostly is room temperature
(15～20 DEG C) liquid storage seminal fluid, the motility of sperm higher (0.6～0.8) during breeding, the conception rate of breeding sow are higher
(75%～85%), but liquid seminal fluid one is can not to preserve for a long time (storage life is typically at 3～5 days), and two is to be not easy at a distance
Transport, therefore, it is impossible to give full play to the Seed practical value of the most top breeding boar of outstanding breeding boar.
In recent years, the research emphasis of swine artificial insemination is gradually transferred on frozen semen.Pig frozen semen technology refers to profit
Make cooling source with dry ice (-79 DEG C), liquid nitrogen (-196 DEG C) etc., by freezing after seminal fluid special handling, be saved in ultralow temperature (-79 DEG C
Or-196 DEG C) state under, reach the purpose preserved for a long time, and semen deposition reach the technology of higher conception rate after thawing.China
It is first to raise pigs and Carnis Sus domestica big producing country in the world, in recent years, annual live pig livestock on hand 4.5～4.7 hundred million, wherein, can numerous mother
Pig livestock on hand 0.48～0.51 hundred million, live pig delivers 6.5～7.1 hundred million for sale, and Carnis Sus domestica always produces 4800～54,000,000 tons, accounts for the world and raises pigs
Total half produced；The most in recent years, the pig industry of China from tradition extensive mode of raising scattered to scale intensive farm
The transition of mode, grow the fast long lean meat species western pig breeds such as white, DABAI, Duroc and have become as the main flow product that large-scale pig farm is raised
Kind, the Artificial Insemination Technology of Swine taken as the leading factor with room temperature liquid storage seminal fluid technology by large-area promotion and application to raising pigs
In production practices.But in the Swine Production of China, the work carried out with application is studied for frozen semen and is short of the most very much,
Therefore, carry out research and the application of technique in a deep going way, not only for the advantage playing technology of artificial insemination further, there is weight
Want meaning, and for carrying out the genetic improvement of boar and for the aspect such as preservation of some peculiar local pig resources in imminent danger
Also there is highly important strategic importance.
Summary of the invention
It is an object of the invention to for above-mentioned technology, it is provided that a kind of quality being effectively improved Cryopreservation of Boar Semen and effect
The freezing of the pig semen of fruit and defreezing method.
For solving the problems referred to above, the technical solution adopted in the present invention is:
The freezing of a kind of pig semen and defreezing method, including the freezing method of pig semen, the defreezing method of pig semen and
The detection of pig semen, it is characterised in that the freezing method of described pig semen comprises the following steps:
Step one: gather outstanding breeding boar seminal fluid and be measured its density, is then diluting this essence with diluent
Liquid, the temperature conditions of dilution seminal fluid is 37.5-40 DEG C, and the ratio of be diluted to finally to live sperm count and boar semen is 0.50-
0.51∶1。
Step 2: the seminal fluid after dilution in step one is stood 0.5-1h under the room temperature environment of 17.5-19.5 DEG C.
Step 3: the seminal fluid in step 2 is moved into the temperature conditions borehole cooling balance 3h of 14-15 DEG C.
Step 4: take the seminal fluid 800g centrifugal treating at a temperature of 14-15 DEG C after the cooling in step 3.
Step 5: be initially charged the diluent I without glycerol, after then making temperature be slowly dropped to 5 DEG C of balance 2h, adds containing sweet
Oil dilution liquid II, then carries out subpackage with tubule.
Step 6: taking the seminal fluid that in step 5, subpackage is good and proceed by freezing, seminal fluid local environment initial temperature is 5 DEG C,
Drop to-5 DEG C with the speed of 3 DEG C/min from 5 DEG C afterwards, under the conditions of-5 DEG C, keep 1min crystallization, then with 50 DEG C/min's
Speed drops to-140 DEG C from-5 DEG C, finally seminal fluid is put into stifling at wide-mouth liquid nitrogen container and liquid nitrogen surface distance 5cm after put into liquid
Nitrogen preserves.
Further, the defreezing method of described pig semen is as follows: take out rapidly the freezing pig semen preserved in liquid nitrogen,
Temperature rises to-5 DEG C with the speed of 50 DEG C/min from-140 DEG C, then rises to 5 DEG C with the speed of 3 DEG C/min from-5 DEG C, thaws.
Further, the detection of described pig semen: take the pig semen after defrosting, by electronic biologic microscope and counting chamber
Its motility of sperm, abnormal rate, effective sperm count are detected.
Preferably, the diluent formulation described in described boar semen method step one is D-Glucose 37.15g, lemon
Lemon acid trisodium 6.00g, EDETATE SODIUM salt 1.25g, sodium bicarbonate 1.25g, potassium chloride 0.75g, penicillin sodium 0.60g, sulphuric acid strepto-
Element 1.00g or D-Glucose 60.00g, trisodium citrate 3.70g, EDETATE SODIUM salt 3.70g, sodium bicarbonate 1.20g, sulphuric acid strepto-
Element 0.50g, penicillin sodium 500,000 unit.
Preferably, the formula of the diluent I in described boar semen method step five and diluent II is: D-Glucose
11.5g, trisodium citrate 11.65g, EDETATE SODIUM salt 2.35g, potassium dihydrogen phosphate 1.75g, streptomycin sulfate 0.5g, polyvinyl alcohol
(PVP.Type11) 1.0g, three carboxymethylamino methanol (Tris) 5.5g, citric acid 4.1g, half dish propylhomoserin 0.07g, OEP (ammonia
Base-sodium laurilsulfate) 0.4g, SDS (sodium lauryl sulphate) 0.3g, PGF2a 0.5g, vitamin E 0.5g, GSH
(glutathion) 0.5g, hyaluronic acid 0.3g, N-acetyl group-D-aminoglucose 0.5g.
Showing through experimental data, after using method freeze-thaw Boar spermatozoa provided by the present invention, sperm is in freezing, defrosting
After vigor reach more than 0.5, the abnormal spermium rate such as deformity is not higher than 19%, the conception rate of the sow that oestruses reach 66% with
On, the average normal litter size of every sow is 9, has reached the technical merit of production application.Show through production practices, this pig essence
The freezing of liquid and defreezing method can make seminal fluid be preserved for a long time, make the use of seminal fluid not limited, no by time and region
It is spaced by this boar death, loss, semen collection or breeds sterile restriction, being effectively increased the utilization rate of boar, give full play to excellent
The effect of good boar, reduces the cost of live population, prevents the drift of excellent genes and the generation of infectious disease, be beneficial to carry out in a deep going way
The union breeding work of pig, it is thus achieved that bigger genetic progress, at aspect tools such as the local improved seeds of protection and quickening breed improvements
There is important researching value and be widely applied prospect.
Detailed description of the invention
Below in conjunction with specific embodiment, the present invention is done further details of elaboration, but the implementation of the present invention is not
Being confined to the scope represented by embodiment, this embodiment is only limitted to the present invention is described, and is not intended to limit the scope of the present invention.
The freezing of a kind of pig semen and defreezing method, including the freezing method of pig semen, the defreezing method of pig semen and
The detection of pig semen, the freezing method of its pig semen comprises the following steps:
Step one: gather outstanding breeding boar seminal fluid and be measured its density, is then diluting this essence with diluent
Liquid, the temperature conditions of dilution seminal fluid is 37.5-40 DEG C, and the ratio of be diluted to finally to live sperm count and boar semen is 0.50-
0.51∶1。
Step 2: the seminal fluid after dilution in step one is stood 0.5-1h under the room temperature environment of 17.5-19.5 DEG C.
Step 3: the seminal fluid in step 2 is moved into the temperature conditions borehole cooling balance 3h of 14-15 DEG C.
Step 4: take the seminal fluid 800g centrifugal treating at a temperature of 14-15 DEG C after the cooling in step 3.
Step 5: be initially charged diluent I, after then making temperature be slowly dropped to 5 DEG C of balance 2h, adds glycerinated dilution
Liquid II, then carries out subpackage with tubule.
Step 6: taking the seminal fluid that in step 5, subpackage is good and proceed by freezing, seminal fluid local environment initial temperature is 5 DEG C,
Drop to-5 DEG C with the speed of 3 DEG C/min from 5 DEG C afterwards, under the conditions of-5 DEG C, keep 1min crystallization, then with 50 DEG C/min's
Speed drops to-140 DEG C from-5 DEG C, finally seminal fluid is put into stifling at wide-mouth liquid nitrogen container and liquid nitrogen surface distance 5cm after put into liquid
Nitrogen preserves.
The defreezing method of pig semen is as follows: take out rapidly the freezing pig semen preserved in liquid nitrogen, temperature with 50 DEG C/
The speed of min rises to-5 DEG C from-140 DEG C, then rises to 5 DEG C with the speed of 3 DEG C/min from-5 DEG C, thaws.
The detection of pig semen: take the pig semen after defrosting, by electronic biologic microscope and counting chamber to its motility of sperm,
Abnormal rate, effective sperm count detect.
Diluent formulation described in boar semen method step one is D-Glucose 37.15g, trisodium citrate
6.00g, EDETATE SODIUM salt 1.25g, sodium bicarbonate 1.25g, potassium chloride 0.75g, penicillin sodium 0.60g, streptomycin sulfate 1.00g or
D-Glucose 60.00g, trisodium citrate 3.70g, EDETATE SODIUM salt 3.70g, sodium bicarbonate 1.20g, streptomycin sulfate 0.50g, green grass or young crops
Mycin sodium 500,000 unit.
The formula of the diluent I described in boar semen method step five and diluent II is: D-Glucose 11.5g,
Trisodium citrate 11.65g, EDETATE SODIUM salt 2.35g, potassium dihydrogen phosphate 1.75g, streptomycin sulfate 0.5g, polyvinyl alcohol
(PVP.Type11) 1.0g, three carboxymethylamino methanol (Tris) 5.5g, citric acid 4.1g, half dish propylhomoserin 0.07g, OEP (ammonia
Base-sodium laurilsulfate) 0.4g, SDS (sodium lauryl sulphate) 0.3g, PGF2a 0.5g, vitamin E 0.5g, GSH
(glutathion) 0.5g, hyaluronic acid 0.3g, N-acetyl group-D-aminoglucose 0.5g.
After diluting boar semen, the process of cooling is the adaptation process that sperm metabolism reduces, when needing the freezing balance of a few hours
Between, but the electrolysis that the slowest rate of cooling can make solution is gradually increased, and causes spermatid membrane lipoprotein complex to destroy and film divides
Solving, when cell membrane syneresis reaches critical minimum volume, cell permeability of the membrane produces irreversible damage, originally can not
Solute through film becomes permeability, in turn results in cell injury or death, and the present invention draws freezing in practical operation
Time is advisable with 2.5～4.0h, and the diluent prepared is age overnight at 4 DEG C, can reach more preferable dilution refrigeration effect.
Spermatid in refrigerating process not only to have tolerance to the low temperature of the state of preservation, and to be amenable to from penetrating
During essence, organism temperature the most excessively reduces vigor to the change of ultra low temperature when preserving, and the latter more seems important than the former.Often
Planting cell has it the most freezing and Thawing Rate requirement, and when freezing rate is too slow, cell to be preserved is easily molten by high concentration
Solve the impact of liquid and dead or suffer to destroy (claiming lysate effect)；When freezing rate is too fast, intracellular fluid is easily caused icing
Phenomenon.The present invention is during boar semen, and in time dropping to-5 DEG C for 5 DEG C, rate of temperature fall is 3 DEG C/min, and this speed will not be right
The serous coat of sperm causes the biggest destruction, drops to-140 DEG C from-5 DEG C, and rate of temperature fall uses 50 DEG C/min, and under the conditions of being somebody's turn to do, sperm can
Quickly through dangerous warm area (0～-60 DEG C), to improve and to freeze the survival rate after essence is thawed.
Principle when pig semen of the present invention thaws is the most contrary with freezing, theoretically, and the situation that freezing/cooldown rate is fast
Under, Thawing Rate is accelerated the most accordingly, and its degree regards dissimilar cell and slightly difference.Quick-thawing phase relatively slower is thawed to essence
The recovery of sub-vigor is favourable, and the time that sperm contacts with concentration solution and cryoprotective agent is shorter, and intracellular external enwergy reaches quickly
The vigor of sperm is made to be recovered to balance.
The present invention is cheap, easily promote, and defreezing method is simple, is more easy to carry out artificial insemination, is suitable for scale
Produce.
Described above to the disclosed embodiments, makes professional and technical personnel in the field be capable of or uses the present invention,
Multiple amendment to embodiment will be apparent from for those skilled in the art, as defined herein typically
Principle can realize without departing from the spirit or scope of the present invention in other embodiments.Therefore, the present invention will not
Can be restricted to embodiment illustrated herein, and be to fit to consistent with principles disclosed herein and features of novelty the widest
Scope.
Claims (5)
1. the freezing of pig semen and a defreezing method, including the freezing method of pig semen, the defreezing method of pig semen and pig
The detection of seminal fluid, it is characterised in that the freezing method of described pig semen comprises the following steps:
Step one: gather outstanding breeding boar seminal fluid and be measured its density, is then diluting this seminal fluid with diluent, dilute
The temperature conditions releasing seminal fluid is 37.5-40 DEG C, and the ratio of be diluted to finally to live sperm count and boar semen is 0.50-0.51: 1.
Step 2: the seminal fluid after dilution in step one is stood 0.5-1h under the room temperature environment of 17.5-19.5 DEG C.
Step 3: the seminal fluid in step 2 is moved into the temperature conditions borehole cooling balance 3h of 14-15 DEG C.
Step 4: take the seminal fluid 800g centrifugal treating at a temperature of 14-15 DEG C after the cooling in step 3.
Step 5: be initially charged the diluent I without glycerol, after then making temperature be slowly dropped to 5 DEG C of balance 2h, adds glycerinated
Diluent II, then carries out subpackage with tubule.
Step 6: taking the seminal fluid that in step 5, subpackage is good and proceed by freezing, seminal fluid local environment initial temperature is 5 DEG C, afterwards
Drop to-5 DEG C with the speed of 3 DEG C/min from 5 DEG C, under the conditions of-5 DEG C, keep 1min crystallization, then with the speed of 50 DEG C/min
Drop to-140 DEG C from-5 DEG C, finally seminal fluid is put into stifling at wide-mouth liquid nitrogen container and liquid nitrogen surface distance 5cm after put into liquid nitrogen and protect
Deposit.
The freezing of a kind of pig semen the most according to claim 1 and defreezing method, it is characterised in that the solution of described pig semen
Freeze method as follows: taking out rapidly the freezing pig semen preserved in liquid nitrogen, temperature rises from-140 DEG C with the speed of 50 DEG C/min
To-5 DEG C, then rise to 5 DEG C with the speed of 3 DEG C/min from-5 DEG C, thaw.
The freezing of a kind of pig semen the most according to claim 1 and defreezing method, it is characterised in that the inspection of described pig semen
Survey as follows: take the pig semen after defrosting, by electronic biologic microscope and counting chamber to its motility of sperm, abnormal rate, effectively essence
Subnumber detects.
The freezing of a kind of pig semen the most according to claim 1 and defreezing method, it is characterised in that described boar semen
Diluent formulation described in method step one is: D-Glucose 37.15g, trisodium citrate 6.00g, EDETATE SODIUM salt 1.25g,
Sodium bicarbonate 1.25g, potassium chloride 0.75g, penicillin sodium 0.60g, streptomycin sulfate 1.00g or D-Glucose 60.00g, Fructus Citri Limoniae
Acid trisodium 3.70g, EDETATE SODIUM salt 3.70g, sodium bicarbonate 1.20g, streptomycin sulfate 0.50g, penicillin sodium 500,000 unit.
The freezing of a kind of pig semen the most according to claim 1 and defreezing method, it is characterised in that described boar semen
Diluent I and the formula of diluent II in method step five be: D-Glucose 11.5g, trisodium citrate 11.65g, EDETATE SODIUM
Salt 2.35g, potassium dihydrogen phosphate 1.75g, streptomycin sulfate 0.5g, polyvinyl alcohol (PVP.Type11) 1.0g, three carboxymethylaminos
Methanol (Tris) 5.5g, citric acid 4.1g, half dish propylhomoserin 0.07g, OEP (amino-sodium laurilsulfate) 0.4g, SDS (ten
Sodium dialkyl sulfate) 0.3g, PGF2a 0.5g, vitamin E 0.5g, GSH (glutathion) 0.5g, hyaluronic acid 0.3g, N-second
Acyl group-D-aminoglucose 0.5g.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201610374193.6A CN106070184A (en) | 2016-05-31 | 2016-05-31 | The freezing of a kind of pig semen and defreezing method |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201610374193.6A CN106070184A (en) | 2016-05-31 | 2016-05-31 | The freezing of a kind of pig semen and defreezing method |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN106070184A true CN106070184A (en) | 2016-11-09 |
Family
ID=57230389
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201610374193.6A Pending CN106070184A (en) | 2016-05-31 | 2016-05-31 | The freezing of a kind of pig semen and defreezing method |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN106070184A (en) |
Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN107535481A (en) * | 2017-09-29 | 2018-01-05 | 界首市国启家庭农场 | A kind of boar semen protective agent |
| CN108378019A (en) * | 2018-03-13 | 2018-08-10 | 洛阳轩智生物科技有限公司 | A kind of frozen stock solution of people's stem spermatogonium |
| CN110786315A (en) * | 2019-10-10 | 2020-02-14 | 北京田园奥瑞生物科技有限公司 | Batch production process of frozen pig semen |
| CN114651815A (en) * | 2022-04-12 | 2022-06-24 | 江苏农牧科技职业学院 | Method for preserving frozen boar semen |
Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| EP2301333A1 (en) * | 2002-07-22 | 2011-03-30 | Xy, Llc | Non-human sperm cell process system |
| 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 |
| CN104322485A (en) * | 2014-10-10 | 2015-02-04 | 山东鑫基牧业有限公司 | Boar frozen semen diluted powder |
-
2016
- 2016-05-31 CN CN201610374193.6A patent/CN106070184A/en active Pending
Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| EP2301333A1 (en) * | 2002-07-22 | 2011-03-30 | Xy, Llc | Non-human sperm cell process system |
| 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 |
| CN104322485A (en) * | 2014-10-10 | 2015-02-04 | 山东鑫基牧业有限公司 | Boar frozen semen diluted powder |
Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN107535481A (en) * | 2017-09-29 | 2018-01-05 | 界首市国启家庭农场 | A kind of boar semen protective agent |
| CN108378019A (en) * | 2018-03-13 | 2018-08-10 | 洛阳轩智生物科技有限公司 | A kind of frozen stock solution of people's stem spermatogonium |
| CN110786315A (en) * | 2019-10-10 | 2020-02-14 | 北京田园奥瑞生物科技有限公司 | Batch production process of frozen pig semen |
| CN114651815A (en) * | 2022-04-12 | 2022-06-24 | 江苏农牧科技职业学院 | Method for preserving frozen boar semen |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN104322484A (en) | Freezing and unfreezing method for boar semen | |
| Maria et al. | Effects of cooling and freezing on sperm motility of the endangered fish piracanjuba Brycon orbignyanus (Characiformes, Characidae) | |
| CN106070184A (en) | The freezing of a kind of pig semen and defreezing method | |
| CN102657149B (en) | Ultra-high activity tubule semen freezing method for sheep | |
| Abdelhakeam et al. | Studies on the presence and absence of glycerol in unfrozen and frozen ram semen: Fertility trials and the effect of dilution methods on freezing ram semen in the absence of glycerol | |
| Alcay et al. | Royal jelly supplemented soybean lecithin-based extenders improve post-thaw quality and incubation resilience of goat spermatozoa | |
| Peruma et al. | Role of reduced glutathione in improving post-thawed frozen seminal characters of poor freezable Jersey crossbred bull semen | |
| CN103918642A (en) | Boar semen refrigerating method and dedicated refrigerating fluid thereof | |
| Golshahi et al. | Disaccharide supplementation of extenders is an effective means of improving the cryopreservation of semen in sturgeon | |
| Cobo et al. | Vitrification of human mature oocytes in clinical practice | |
| CN107683850A (en) | A kind of swamp eel sperm super-low temperature freezing preserves liquid and its Cryopreservation method | |
| CN103931531A (en) | Method for long-term storage of crassostrea hongkongensis eyespot larvas by critical low temperatures before freezing point | |
| CN103262837B (en) | Livestock seminal fluid cryoprotectant and application thereof | |
| Araújo-Lemos et al. | Comparison of vitrification and conventional freezing for cryopreservation of caprine embryos | |
| Bahrawy et al. | Effect of collection frequency, extender and thawing temperature on the motility recovery of cryopreserved dromedary camel spermatozoa | |
| CN104304236A (en) | Erythroculter ilishaeformis semen cryopreservation solution | |
| Dalal et al. | Comparison of three programmable freezing protocols for the cryopreservation of buffalo bull semen | |
| Kozdrowski | The effect of trehalose on post-thaw viability and fertility of European brown hare (Lepus europaeus Pallas, 1778) spermatozoa | |
| Widyaningsih et al. | Effect of various concentrations of palm date (Phoenix dactylifera L.) on spermatozoa motility of giant grouper (Epinephelus lanceolatus) | |
| CN102870763A (en) | Cryopreservation method of Lutjanus erythropterus sperms | |
| CN102986652B (en) | Superfreeze preservation method of pink salmon sperm | |
| Gerzilov | Influence of various cryoprotectants on the sperm mobility of Muscovy semen before and after cryopreservation. | |
| da Silva | Artificial insemination and cryopreservation of boar semen: Current state and problematics | |
| Patel et al. | Effect of semen diluent additives on spermatozoal viability of Kankrej bull semen following cryopreservation | |
| CN110226592A (en) | A kind of flat Rockfish sperm cryopreservation method of Xu Shi |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| C10 | Entry into substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| RJ01 | Rejection of invention patent application after publication | ||
| RJ01 | Rejection of invention patent application after publication |
Application publication date: 20161109 |