CN101554152A - Frozen semen diluent and preparation method thereof - Google Patents
Frozen semen diluent and preparation method thereof Download PDFInfo
- Publication number
- CN101554152A CN101554152A CNA2009100226257A CN200910022625A CN101554152A CN 101554152 A CN101554152 A CN 101554152A CN A2009100226257 A CNA2009100226257 A CN A2009100226257A CN 200910022625 A CN200910022625 A CN 200910022625A CN 101554152 A CN101554152 A CN 101554152A
- Authority
- CN
- China
- Prior art keywords
- milligrams
- milliliters
- vitamin
- seminal fluid
- dilution
- 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.)
- Granted
Links
Landscapes
- Medicines Containing Material From Animals Or Micro-Organisms (AREA)
- Agricultural Chemicals And Associated Chemicals (AREA)
Abstract
The invention discloses a novel frozen semen diluent used for improving the freeze-thaw quality and fertilization rate of frozen bovine semen and a preparation method thereof. Every 100ml of the prepared frozen semen diluent contains 0.9-1.2g of sodium citrate, 3-5g of sucrose, 18-22ml of yelk, 4-7ml of glycerin, 6-9mg of vitamin E, 500-700mg of vitamin C, 35mg of spectinomycin and 80 thousand units of gentamicin. The invention adds VE and VC into the semen diluent for the first time, so as to increase the motility rate and acrosome integrity of the frozen sperm as well as the activity of antioxidant enzyme in seminal plasma, thus improving the quality and fertilization rate of the freeze-thaw sperm.
Description
Technical field
The present invention relates to and belongs to the test-tube improving technology category of ox, relates in particular to a kind of cow frozen semen that is used for and improves freezing seminal fluid dilution and the compound method thereof that Niu Jingzi freezes the back quality.
Background technology
The freezing of livestock semen successfully produced extremely far-reaching influence for the development of people worker's insemination techniques, at first it utilizes the time to lay a good foundation for availability and the prolongation that enlarges outstanding sire, and secondly the development for artificial insemination and cryobiology provides wide space.Because frozen semen is more convenient in transportation, no matter from the time or from the space, the breeding efficiency of sire is not fully exerted, and has quickened the paces of domestic animal breed improvement.Though each state has all launched the research to frozen semens such as ox, pig, horse, chickens, has only the technology of artificial insemination of cow frozen semen to be widely used in production practices now.Aspect the dilution development of domestic animal frozen semen, the main effect of all dilutions and preservation environment all is in order to provide enough nutriments to supply sperm metabolism needs to sperm, prevent the rapid freezing damage that causes of sperm, cushion to prevent the variation of pH, keep suitable osmotic pressure, equilibrium electrolyte, suppress the growth of bacterium, enlarge extension rate with increase semen deposition number of times, and the raising sperm freezes back motility rate, the utilization ratio of raising breeding stock and seminal fluid.
In recent years, along with Chinese economic development, the raising of living standards of the people, milk has been come into huge numbers of families gradually, so the quantity of milk cow is in recent years in the violent speed development of China.Chinese elite high yield cow improved variety degree is not high, and population size is little, and yielding poorly down becomes the factor of the restriction that milk cow develops.So with the frozen semen of the high bull that gives milk the low milking cow of China being improved becomes the task of top priority.
In order to improve constantly the refrigerating effect of milk cow frozen semen, in the past more than 60 year, the expert is making great efforts to explore and seek optimum cryoprotector always both at home and abroad, improves the refrigerating effect of milk cow and beef cattle frozen semen.Studies show that both at home and abroad seminal fluid is handled back sperm DNA damage aggravation through freeze-thaw, cause that sperm quality and fertilization rate reduce after the sperm freeze thawing, this is a problem of puzzlement sperm fertilizing ability always.Therefore the height of frozen semen quality has become to restrict the key factor of milk cow, beef cattle artificial insemination success rate.But in production practices, the sperm freeze thawing is arranged often after vigor not high, fertilization rate is low, influences test-tube effect.Therefore, according to present trend, utilize breadboard condition that the sperm dilution formula of liquid of milk cow is studied, the semen diluent that exploitation makes new advances, so both can improve survival rate and fertilization rate after the sperm freeze thawing, and accelerate the improvement dynamics of China milk cow again, be improvement and the milk yield that improves milk cow rapidly, increase the economic benefit of dairy, pulling place and regional economy development have positive effect.
Below be the relevant references that the applicant retrieves:
[1]Partida?S?L?G，Setchell?B?P，Maxwell?W?M.Epididymal?compoundsand?antioxidants?in?diluents?for?the?frozen?storage?of?ramspermatozoa[J].Reprod?Fertil?Dev，1997，9(7)：689-696。
[2]Foote?R?H，Brockett?C?C，Kaproth?M?T.Motility?and?fertility?ofbull?sperm?in?whole?milk?extender?containing?antioxidants[J].AnimReprod?Sci，2002，71(1-2)：13-23。
[3]Jones?R，Mann?T.Damage?to?ram?spermatozoa?by?peroxidantion?ofendogenous?phospholipids[J].J?Reprod?Fertil，1997，50(2)：261-26。
[4] Sun Shuqin, Cai Jiguang, Wang Lige etc. add the influence [J] of the cobalamin of variable concentrations to ox tubule frozen semen sperm morphology in the dilution.Journal 2004,6 (2) 10-13 of Liaoning agricultural occupation technical college.
[5] Ha Fu, Zhao Youzhang adds vitamin B compound and improves and freeze the essence back Study on Quality [J] of thawing in the sheep freezing seminal fluid dilution. Gansu Agriculture University,'s journal, 2003,3 (1): 17-19,46.
[6] Luo Hailing, Jia Zhihai, Zhu Shien etc. add the influence [J] of vitamin E to sheep frozen semen quality in the dilution. Chinese herding magazine, 2004,40 (11): 7-10.
[7]B.A.Ball，V.Medina，C.G.Gravance，I.Baumber.Effect?ofantionxidants?on?presernation?of?motity?viability?and?acrosomalintegrity?of?equine?spermatozoa?during?storage?at?5℃，Theriogenology2001，56：577-569。
[8]Nabil?Mansour，Mary?A.McNiven，Gavin?F.Richardson.The?effect?ofdietary?supplementation?with?blueberry，a-tocopherol?or?astaxanthinon?oxidative?stability?of?Arctic?char(Salvelinus?alpinus)semen.Theriogenology?66(2006)373-382。
[9] Luo Hailing, Jia Zhihai, Zhu Shien, vitamin E improves the mechanism research that the sheep particle freezes elaboration matter, Chinese plant-eating animal 2005,21 (8) 21-23.
[10]M.Moussa?V.Martinet，A.Trimeche，D.Tainturier?and?M.Anton?Lowdensity?lipoproteins?extracted?from?hen?egg?yolk?by?an?easy?method：cryoprotect?ive?effect?on?frozen-thawed?bull?semen?Theriogenology?57(2002)1695-1706。
[11]Talevi?R，Pelosi?S，Sansone?G，et?al.Effects?of?differentpre-freezing?rates?on?buffalo?sperm?Motility?and?ultrastructurepreservation[J].Buffalo.1999(43)：537-539。
[12]Mariola?Slowinska，Halina?Karol，Andrzej?Ciereszko.Comet?assayof?fresh?and?cryopreserved?bull?spermatozoa.Cryobiology?56(2008)100-102。
[13]Marti?a，L.Mara?b，J.I.Marti?c，T.Seasonal?variations?inantioxidant?enzyme?activity?in?ram?seminal?plasma?E?[J].Theriogenology?67(2007)1446-1454。
[14]Ukeda?H，Maeda?S，Ishii?T，Sawamura?M.Spectrophotometric?assayfor?superoxide?dismutase?based?on?tetrazolium?salt3’-{1-[(Phenylamino)-carbonyl]-34-tetrazolium}-bis(4-methoxy-6-nitro)benzenesulphonic?acid?hydrate?reduction?by?xanthinexanthineoxidase.Anal?Biochem?1997；251：206-209。
[15] Li Xiaodong, Bian Fengyu, fourth can be etc., and ox is frozen the experimental study of smart dilution prescription, Chinese milk cow, 1998 (3): 15～17.
[16] Wang Xiaoke, Yang Jian influences the factor [J] of domestic animal frozen semen quality, Inner Mongol herding science, 2002,6:20～21.
[17]Medearos?CMO，Forell?F，Oliveira?ATD，et?al.Current?status?ofsperm?crypereseration：Why?isn’t?it?better？[J].Theriogenology，2002，57：327～344。
[18] Wang Lihong, Wang Shouxun, Zhang Wei, the bull semen detection method compares, Chinese milk cow, 2002 (3): 45～48.
[19] Zhang Zhongcheng, livestock thremmatology [M]. Beijing: Chinese agriculture publishing house, 2004.
[20]Aitken?R?J，Clarkson?J?S.Cellular?basis?of?defective?spermfunction?and?its?association?with?the?genesis?of?reactive?oxygenspecies?by?human?spermatozoa[J].J?Reprod?Fertil，1998，81：459-469。
[21] cow frozen semen quality of production administrative manual (trying), national livestock husbandry veterinarian general station, cow frozen semen quality supervision and test test center of the Ministry of Agriculture, 2000,21～23.
[22] Liu Yutang, the analysis of Influential Factors [J] that semen freezing is preserved. the Heilungkiang animal produces magazine, 2003,11 (2): 10～14.
[23] Qiu Dianrui influences the factor [J] of cow frozen semen quality. Chinese herding magazine, 2002,35 (6): 49～50.
Summary of the invention
Defective or deficiency at above-mentioned prior art exists the objective of the invention is to, and a kind of novel freezing seminal fluid dilution and compound method thereof that is used to improve quality and fertilization rate after the cow frozen semen freeze thawing is provided.This freezing seminal fluid dilution that makes can obviously improve the vigor that sperm freezes antioxidant enzyme in back motility rate and acrosomal integrity and the refining, thereby improves quality and fertilization rate after the sperm freeze thawing.
In order to realize above-mentioned task, the present invention takes following technical solution:
A kind of freezing seminal fluid dilution, it is characterized in that, contain in per 100 milliliters of this freezing seminal fluid dilution that makes: sodium citrate: 0.9 gram～1.2 grams, sucrose: 3 grams～5 grams, yolk: 18 milliliters～22 milliliters, glycerine: 4 milliliters～7 milliliters, vitamin E: 6 milligrams～9 milligrams, vitamin C: 500 milligrams～700 milligrams, spectinomycin: 35 milligrams, gentamicin 80,000 units (IU).
The compound method of above-mentioned freezing seminal fluid dilution is characterized in that, comprises the following steps:
Accurately take by weighing sodium citrate, sucrose, yolk, vitamin E, vitamin C and glycerine in claim 1 prescription, adding distil water to 100 milliliter stirs on magnetic stirring apparatus, obtains mixed solution, puts into 4 ℃ of refrigerator overnight then;
Before the sperm dilution, in mixed solution, add spectinomycin milligram and gentamicin in claim 1 prescription, and stirred 10 minutes;
The acid-base value of adjusting semen diluent with acid or alkali is carried out the bacteriological filtration processing with 0.22 micron filter to dilution between 6.7～7.2, obtain freezing seminal fluid dilution;
During use, with 0.22 micron filter freezing seminal fluid dilution is carried out bacteriological filtration and handle, it is standby to put into 37.5 ℃ of water-baths after finishing, and promptly can be used for thawing of seminal fluid according to conventional seminal fluid preparation method then.
The present invention is first V
E, V
CUnite and joined in the semen diluent; and the compound method of dilution revised; experimental data shows; freezing freezing seminal fluid dilution of the present invention has not only improved vigor and the perforatorium percentage of head rice after the sperm freeze thawing; and improved vigor after the freeze thawing of the catalase (CAT) that is used for protecting sperm fertilizing ability in the refining, glutathione peroxidase (GR), glutathione reductase (GSH), guaranteed that more this seminal fluid prescription can improve sperm and freeze the back quality.
Embodiment
According to technical scheme of the present invention, the material of preparing 100 milliliters freezing seminal fluid dilution needs is: sodium citrate: 0.9～1.2 gram, sucrose: 3～5 grams, yolk: 18～22 milliliters, glycerine: 4～7 milliliters, vitamin E: 6～9 milligrams, vitamin C: 500～700 milligrams, spectinomycin: 35 milligrams, gentamicin: 80,000 units (IU).
Its best freezing seminal fluid dilution prescription is: the content of material that contains in per 100 milliliters is: sodium citrate: 1.073 milligrams, sucrose: 4.344 milligrams, yolk: 20 milliliters, glycerine: 6 milliliters, vitamin E: 8 milligrams, vitamin C: 600 milligrams, spectinomycin: 35 milligrams, gentamicin 80,000 units (IU).
Its compound method is:
1) takes by weighing raw material in the prescription: sodium citrate, sucrose, yolk, glycerine, vitamin E, vitamin C, distilled water.
2) with above sodium citrate, sucrose, yolk, vitamin C, vitamin E, glycerine is configured to basal liquid with distilled water, uses magnetic stirrer 60 minutes, obtains mixed solution, and it is standby to spend the night in 4 ℃ of refrigerators.
3) before the sperm dilution, in mixed solution, add the gentamicin and the spectinomycin of formula ratio, then magnetic stirrer 10 minutes.
4) adjusting the dilution pH value with acid or alkali is 6.5～7.5, with 0.22 micron filter dilution is carried out bacteriological filtration and handles, and obtains semen diluent.
Freezing seminal fluid dilution and the conventional formulation of measuring the present embodiment preparation are being frozen back motility rate, acrosomal integrity, refining antioxidant enzyme to the ox sperm.Wherein the refining antioxidant enzyme of Ce Dinging mainly comprises catalase (CAT), glutathione peroxidase (GR), glutathione reductase (GSH).
The detection method of sperm motility rate, acrosomal integrity and refining antioxidant enzyme is as follows:
(1) under 35 ℃～37 ℃ temperature, carries out the motility rate inspection immediately behind the semen thawing.During evaluation, dip in glass bar and to get 1 seminal fluid, drip on slide, add cover glass, make between cover glass and the slide to be full of seminal fluid, place observation 400 times～600 power microscopes under then.In field of microscope, by seminal fluid upper, middle and lower-ranking liquid level sperm motility information summary being observed the evaluation sperm motility rate.
(2) get a seminal fluid after thawing and drop on the clean slide, be 45 degree, seminal fluid is smeared to opposite side, make and smear sheet with another slide.Fix 15 minutes with 1～2 milliliter of formalin phosphate buffer behind the natural air drying, dyeed 90 minutes with Ji's nurse Sa behind the washing and drying, remove the dye liquor of smearing on the sheet then, air-dry being placed on observed 300 sperms, record sperm sum and the complete number of sperm of acrosome under 1000 times of biomicroscopes.
(3) vigor of refining antioxidant enzyme is measured in strict accordance with operational procedure with the related kit of measuring enzyme (kit of measuring enzyme all builds up biotech company from Nanjing).
Below be the concrete experimental example that the inventor provides, the adult He Sitan stock bull that test is selected the physique stalwartness for use, is highly sexed is raised by stock bull feeding and management requirement, and its semen collection frequency is 3 times weekly.Utilize the artificial vagina acquisition method to gather bull semen, after the semen collection, immediately at 35 ℃～37 ℃ following microscopies.The seminal fluid of being gathered is milky or milk yellow, free from extraneous odour, and sperm morphology is normal, and motility rate reaches and can be used to make frozen semen more than 0.7.
This test is provided with three different freezing seminal fluid dilution prescription and conventional formulation:
Prescription 1: prepare 100 milliliters of dilutions: sodium citrate: 1.073 milligrams, sucrose: 4.344 milligrams, yolk: 20 milliliters, glycerine: 6 milliliters, vitamin E: 8 milligrams, vitamin C: 600 milligrams, spectinomycin: 35 milligrams, gentamicin: 80,000 units (IU).
Prescription 2: prepare 100 milliliters of dilutions: sodium citrate: 1.073 milligrams, sucrose: 4.344 milligrams, yolk: 20 milliliters, glycerine: 6 milliliters, vitamin C: 600 milligrams, spectinomycin: 35 milligrams, gentamicin: 80,000 units (IU).
Prescription 3: prepare 100 milliliters of dilutions: sodium citrate: 1.073 milligrams, sucrose: 4.344 milligrams, yolk: 20 milliliters, glycerine: 6 milliliters, vitamin E: 8 milligrams, spectinomycin: 35 milligrams, gentamicin: 80,000 units (IU).
Conventional formulation: prepare 100 milliliters of dilutions: sodium citrate: 1.073 milligrams, sucrose: 4.344 milligrams, yolk: 20 milliliters, glycerine: 6 milliliters, spectinomycin: 35 milligrams, gentamicin: 80,000 units (IU).
Strictness takes by weighing ingredient by above prescription: sodium citrate, sucrose, yolk, glycerine, vitamin E and vitamin C are settled to 100 milliliters with distilled water; Stirring stirred in 60 minutes on magnetic stirring apparatus, obtained mixed solution, put into 4 ℃ of refrigerator overnight then.
Before the sperm dilution, in mixed solution, add 35 milligrams of spectinomycins, gentamicin 80,000 units (IU), and stirred 10 minutes, with the acid-base value of acid or alkali adjustment dilution, decide dilution PH between 6.7～7.2 with the PH instrumentation, obtain semen diluent.
During use, with 0.22 micron filter dilution is carried out bacteriological filtration and handle, insert after finishing in 37.5 ℃ of water-baths, can carry out the dilution of thawing of frozen semen then according to conventional seminal fluid preparation method, semen quality is detected in the back that thaws, and specific targets see the following form:
The different formulations freezing seminal fluid dilution to freeze thawing after the influence (mean ± standard deviation) of sperm quality
| Handle | Prescription one | Prescription two | Prescription three | Conventional formulation |
| Motility rate (%) | 39.33±0.58a | 37.33±0.58b | 38.67±1.53ab | 35.33±1.15c |
| Acrosomal integrity (%) | 76.33±2.08a | 74.33±1.53ac | 75.67±1.53a | 71.67±1.53bc |
| Activity of catalase (U/mL) | 4.57±0.50a | 3.99±0.12b | 4.37±0.1ab | 3.97±0.21b |
| Glutathione peroxidase activity (U/L) | 100.53±1.76a | 48.48±2.93d | 80.14±1.10b | 74.76±1.56c |
| Glutathione reductase vigor (U/L) | 136.67±2.06b | 121.83±1.72c | 165.98±1.73a | 117.51±1.30c |
Annotate: with the different sign significant difference of institute's marking-up parent phase (P＜0.05) behind the line data, institute's marking-up parent phase is with indicating difference not remarkable (P＜0.05).
As can be seen from the above table, freezing seminal fluid dilution provided by the present invention all significantly is better than other prescriptions (P＜0.05) in indexs such as sperm motility rate, acrosomal integrity, catalase, glutathione peroxidase, glutathione reductase vigor.And fill a prescription one respectively than filling a prescription two or three in raising 5.6%～11.3% aspect the sperm motility rate, improve 11.3% than conventional formulation.Prescription one exceeds other and fills a prescription approximately 3.7%～5.5% aspect acrosomal integrity, is higher than conventional formulation 6.5%.Prescription one exceeds other prescription 0.5%～10.1% aspect activity of catalase, exceeds conventional formulation 15.1%.Exceed conventional formulation 34.4% at prescription one aspect the vigor of glutathione peroxidase, glutathione reductase vigor aspect exceeds 16.31% than traditional common prescription.So draw three prescriptions of the freezing seminal fluid dilution preparation of present embodiment preparation through above analysis, the product mass-energy to sperm in the sperm freezing course of defrosting plays the maximum protection effect, wherein filling a prescription one is best freezing seminal fluid dilution prescription.
Claims (3)
1, a kind of freezing seminal fluid dilution, it is characterized in that, contain in per 100 milliliters of this freezing seminal fluid dilution that makes: sodium citrate: 0.9 gram～1.2 grams, sucrose: 3 grams～5 grams, yolk: 18 milliliters～22 milliliters, glycerine: 4 milliliters～7 milliliters, vitamin E: 6 milligrams～9 milligrams, vitamin C: 500 milligrams～700 milligrams, spectinomycin: 35 milligrams, gentamicin 80,000 units.
2, freezing seminal fluid dilution as claimed in claim 1, it is characterized in that, contain in per 100 milliliters of this freezing seminal fluid dilution that makes: sodium citrate: 1.073 milligrams, sucrose: 4.344 milligrams, yolk: 20 milliliters, glycerine: 6 milliliters, vitamin E: 8 milligrams, vitamin C: 600 milligrams, spectinomycin: 35 milligrams, gentamicin: 80,000 units.
3, the compound method of the described freezing seminal fluid dilution of claim 1 is characterized in that, comprises the following steps:
Accurately take by weighing sodium citrate, sucrose, yolk, vitamin E, vitamin C and glycerine in claim 1 prescription, adding distil water to 100 milliliter stirs on magnetic stirring apparatus, obtains mixed solution, puts into 4 ℃ of refrigerator overnight then;
Before the sperm dilution, in mixed solution, add the spectinomycin milligram and the gentamicin of formula ratio, and stirred 10 minutes;
The acid-base value of adjusting semen diluent with acid or alkali is carried out the bacteriological filtration processing with 0.22 micron filter to dilution between 6.7～7.2, obtain freezing seminal fluid dilution.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN2009100226257A CN101554152B (en) | 2009-05-21 | 2009-05-21 | Frozen semen diluent and preparation method thereof |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN2009100226257A CN101554152B (en) | 2009-05-21 | 2009-05-21 | Frozen semen diluent and preparation method thereof |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN101554152A true CN101554152A (en) | 2009-10-14 |
| CN101554152B CN101554152B (en) | 2012-05-09 |
Family
ID=41172551
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN2009100226257A Active CN101554152B (en) | 2009-05-21 | 2009-05-21 | Frozen semen diluent and preparation method thereof |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN101554152B (en) |
Cited By (13)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101999342A (en) * | 2010-10-08 | 2011-04-06 | 内蒙古蒙牛繁育生物技术股份有限公司 | Novel preparation method of yolk diluent |
| CN102578074A (en) * | 2011-12-22 | 2012-07-18 | 内蒙古赛科星繁育生物技术股份有限公司 | Method for preparing livestock sex control frozen sperm by adding antioxidants of CAT (Catalase) and VE (Vitamin E) |
| CN102726372A (en) * | 2012-07-18 | 2012-10-17 | 尹旭升 | Frozen semen diluent formula for flocks and herds as well as making method |
| CN102907417A (en) * | 2012-10-22 | 2013-02-06 | 西北农林科技大学 | Freezing diluent formula for improving quality of Holstein bull frozen-thawed X-spermatozoa |
| CN103314951A (en) * | 2013-07-19 | 2013-09-25 | 扬州大学 | Method suitable for prolonging normal-temperature preservation time of seminal fluid of adult male rabbit and diluent |
| CN104365583A (en) * | 2014-11-07 | 2015-02-25 | 郜鸿生物科技（上海）有限公司 | Extremely-low sperm freezing protective agent and application thereof |
| 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 |
| CN105794769A (en) * | 2016-04-22 | 2016-07-27 | 李志新 | Special diluting liquid for bovine seminal plasma |
| CN107251892A (en) * | 2017-05-25 | 2017-10-17 | 中国热带农业科学院热带作物品种资源研究所 | It is a kind of suitable for the solution for dilution of cock semen of torrid areas, preparation and its application process |
| CN107396918A (en) * | 2016-11-15 | 2017-11-28 | 青海省家畜改良中心 | It is a kind of to be used to preserve dilution for hiding sheep seminal fluid and preparation method thereof for a long time |
| CN109258625A (en) * | 2018-10-14 | 2019-01-25 | 青海大学 | It is a kind of for improve yak frozen semen freeze after quality dilution formula of liquid |
| CN109329272A (en) * | 2018-11-23 | 2019-02-15 | 北京太东生物科技有限公司 | A kind of spermatozoa cryopreservation liquid and its preparation method and application |
| CN114403131A (en) * | 2022-01-21 | 2022-04-29 | 河北科技师范学院 | Semen cryopreservation liquid and application thereof |
Family Cites Families (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| 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 |
| EP1730266A2 (en) * | 2004-03-29 | 2006-12-13 | Monsanto Technology, LLC | Sperm suspensions for sorting into x or y chromosome-bearing enriched populations |
-
2009
- 2009-05-21 CN CN2009100226257A patent/CN101554152B/en active Active
Cited By (16)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101999342B (en) * | 2010-10-08 | 2012-11-07 | 内蒙古赛科星繁育生物技术股份有限公司 | Novel preparation method of yolk diluent |
| CN101999342A (en) * | 2010-10-08 | 2011-04-06 | 内蒙古蒙牛繁育生物技术股份有限公司 | Novel preparation method of yolk diluent |
| CN102578074A (en) * | 2011-12-22 | 2012-07-18 | 内蒙古赛科星繁育生物技术股份有限公司 | Method for preparing livestock sex control frozen sperm by adding antioxidants of CAT (Catalase) and VE (Vitamin E) |
| CN102726372A (en) * | 2012-07-18 | 2012-10-17 | 尹旭升 | Frozen semen diluent formula for flocks and herds as well as making method |
| CN102907417A (en) * | 2012-10-22 | 2013-02-06 | 西北农林科技大学 | Freezing diluent formula for improving quality of Holstein bull frozen-thawed X-spermatozoa |
| CN103314951A (en) * | 2013-07-19 | 2013-09-25 | 扬州大学 | Method suitable for prolonging normal-temperature preservation time of seminal fluid of adult male rabbit and diluent |
| 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 |
| CN104365583B (en) * | 2014-11-07 | 2016-08-24 | 郜鸿生物科技（上海）有限公司 | A kind of rare sperm freezing protecting agent and application thereof |
| CN104365583A (en) * | 2014-11-07 | 2015-02-25 | 郜鸿生物科技（上海）有限公司 | Extremely-low sperm freezing protective agent and application thereof |
| CN105794769A (en) * | 2016-04-22 | 2016-07-27 | 李志新 | Special diluting liquid for bovine seminal plasma |
| CN107396918A (en) * | 2016-11-15 | 2017-11-28 | 青海省家畜改良中心 | It is a kind of to be used to preserve dilution for hiding sheep seminal fluid and preparation method thereof for a long time |
| CN107251892A (en) * | 2017-05-25 | 2017-10-17 | 中国热带农业科学院热带作物品种资源研究所 | It is a kind of suitable for the solution for dilution of cock semen of torrid areas, preparation and its application process |
| CN109258625A (en) * | 2018-10-14 | 2019-01-25 | 青海大学 | It is a kind of for improve yak frozen semen freeze after quality dilution formula of liquid |
| CN109329272A (en) * | 2018-11-23 | 2019-02-15 | 北京太东生物科技有限公司 | A kind of spermatozoa cryopreservation liquid and its preparation method and application |
| CN114403131A (en) * | 2022-01-21 | 2022-04-29 | 河北科技师范学院 | Semen cryopreservation liquid and application thereof |
| CN114403131B (en) * | 2022-01-21 | 2023-02-24 | 河北科技师范学院 | Semen cryopreservation liquid and application thereof |
Also Published As
| Publication number | Publication date |
|---|---|
| CN101554152B (en) | 2012-05-09 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN101554152B (en) | Frozen semen diluent and preparation method thereof | |
| Thundathil et al. | The use of in vitro fertilization techniques to investigate the fertilizing ability of bovine sperm with proximal cytoplasmic droplets | |
| Roth et al. | Scimitar-horned oryx (Oryx dammah) spermatozoa are functionally competent in a heterologous bovine in vitro fertilization system after cryopreservation on dry ice, in a dry shipper, or over liquid nitrogen vapor | |
| Lone et al. | Effect of transportation temperature on the quality of cauda epididymal spermatozoa of ram | |
| Vick et al. | Improved cryopreservation of domestic cat sperm in a chemically defined medium | |
| Filliers et al. | In vitro evaluation of fresh sperm quality in tomcats: a comparison of two collection techniques | |
| Liu et al. | Effects of pH during liquid storage of goat semen on sperm viability and fertilizing potential | |
| Xu et al. | Semen dilution for assessment of boar ejaculate quality in pig IVM and IVF systems | |
| Santiago-Moreno et al. | Effect of egg yolk concentration on cryopreserving Spanish ibex (Capra pyrenaica) epididymal spermatozoa | |
| Gibb et al. | Recent developments in stallion semen preservation | |
| Krishnakumar et al. | Effect of reproductive seasonality on gamete quality in the North American bison (Bison bison bison) | |
| Tipkantha et al. | Influence of living status (single vs. paired) and centrifugation with colloids on the sperm morphology and functionality in the clouded leopard (Neofelis nebulosa) | |
| Kang et al. | Spermatozoa motility, viability, acrosome integrity, mitochondrial membrane potential and plasma membrane integrity in 0.25 mL and 0.5 mL straw after frozen-thawing in Hanwoo bull | |
| Gororo et al. | Effects of different extenders and storage temperatures on longevity of small East African goat (Capra hircus) semen | |
| de Araujo et al. | Ocelot and oncilla spermatozoa can bind hen egg perivitelline membranes | |
| Gañán et al. | Male reproductive traits, semen cryopreservation, and heterologous in vitro fertilization in the bobcat (Lynx rufus) | |
| Gamboa et al. | Seminal traits, suitability for semen preservation and fertility in the native Portuguese horse breeds Puro Sangue Lusitano and Sorraia: Implications for stallion classification and assisted reproduction | |
| Asr et al. | The evaluations of Tris-citrate acid or Bioxcell extenders on the post-thawed buffalo sperm parameters | |
| Choi et al. | Semen characteristics of genetically identical male cats cloned via somatic cell nucleus transfer | |
| Nitharwal et al. | Effect of season on semen quality parameters in Murrah buffalo | |
| Pérez-Garnelo et al. | Post-thaw viability of European bison (Bison bonasus) semen frozen with extenders containing egg yolk or lipids of plant origin and examined with a heterologous in vitro fertilization assay | |
| Wysokińska et al. | Morphometrical characteristics of spermatozoa in Polish Landrace boars with regard to the number of spermatozoa in an ejaculate | |
| Karunakaran et al. | Semen preservation and artificial insemination in Bengal Goat at field level | |
| CN102907417B (en) | Freezing diluent formula for improving quality of Holstein bull frozen-thawed X-spermatozoa | |
| Gamal et al. | Correlations between semen parameters and conception rate in buffaloes |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| C10 | Entry into substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| C14 | Grant of patent or utility model | ||
| GR01 | Patent grant |