CN102845413A - Diluted rabbit semen preserving fluid - Google Patents
Diluted rabbit semen preserving fluid Download PDFInfo
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- CN102845413A CN102845413A CN2012103612246A CN201210361224A CN102845413A CN 102845413 A CN102845413 A CN 102845413A CN 2012103612246 A CN2012103612246 A CN 2012103612246A CN 201210361224 A CN201210361224 A CN 201210361224A CN 102845413 A CN102845413 A CN 102845413A
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- rabbit semen
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- artificial insemination
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Abstract
The invention discloses a diluted rabbit semen preserving fluid. The diluted rabbit semen preserving fluid comprises the components including 2.13g of citric acid, 3.17g of Tris and 0.60g of anhydrous glucose, and is prepared by dissolving the components into 100ml of distilled water, and then autoclaving, placing the solution into a sterile operation desk after the solution is autoclaved and cooled, and adding 2ml of yolk, 200 thousands IU of penicillin and 200 thousands IU of streptomycin. The test results show that the self-mixed diluted rabbit semen preserving fluid is capable of storing the male rabbit semen for above 60 hours at 4 DEG C in vitro, and the activity is more than 0.5 (the activity of 0.5 is an index which can guarantee normal artificial insemination). The in vitro storage capability of the diluted rabbit semen preserving fluid is far higher than 5% of glucose solution and normal saline. Simultaneously, the pregnancy rate of the artificial insemination test is about 75% when the activity is 0.5, and the test results also show that the artificial insemination can be carried out when the rabbit semen is stored in vitro within 60 hours in the formula.
Description
Technical field
What the present invention relates to is that a kind of rabbit sperm dilution is preserved liquid.
Background technology
Rabbit is Herbivore, is that country promotes the livestock breeding industry restructuring to give priority to the agricultural projects of support at present, also is the specialty industries of present China rural area increase wealth simultaneously.Over nearly 30 years, China's livestock breeding has obtained significant progress, but the rabbit level of industrial technology is compared to other poultry industry and also relatively lags behind, propagation technique aspect particularly, pig, ox, chicken etc. realize that already artificial insemination breeds, but the rabbit industry also adopts the mode of natural mating to breed substantially at present, has seriously restricted the development of rabbit industry.
For a long time, because the propagation technique problem of rabbit does not obtain breakthrough, cause the rabbit productivity effect lower, can not demonstrate fully the large-scale cultivation benefit.Therefore, improve the rabbit propagation technique, adopt artificial insemination, realize that all in all out periodization production model of estrus synchronization, the childbirth same period, the wean same period, the listing same period seems particularly necessary; What do that artificial insemination at first will solve is the external preservation problem of buck seminal fluid, and the research and development that sperm dilution is preserved liquid are again the most important things, and the prerequisite that seminal fluid can effective external preservation is the technology of artificial insemination success also is the guarantee of this technology application simultaneously.
Summary of the invention
Technical problem to be solved by this invention is to provide a kind of rabbit sperm dilution to preserve liquid for the deficiencies in the prior art.
Technical scheme of the present invention is as follows:
A kind of rabbit sperm dilution is preserved liquid, component is: citric acid 2.13g, Tris3.17g, DEXTROSE ANHYDROUS 0.60g is dissolved in the 100ml distilled water, then carrying out autoclaving processes, after the complete cooling subject to sterilization, put it into aseptic operating platform, add 2ml yolk, penicillin 200,000 IU, streptomycin 200,000 IU.
Result of the test shows, the sperm dilution of autogamy is preserved liquid can be more than 4 ℃ of external preservation buck seminal fluid 60h, and vigor is greater than 0.5 (vigor 0.5 is to guarantee normally to carry out test-tube index).It is higher than 5% glucose solution and physiological saline far away in external hold capacity.The conception rate of the artificial insemination test (during vigor 0.5) of carrying out simultaneously is about 75%, and result of the test shows that also this dilution prescription all can carry out artificial insemination at external preservation rabbit seminal fluid 60h with interior.
Description of drawings
Fig. 1 is that autogamy dilution preservation liquid was preserved outward in the seminal fluid 10h time, and vigor is 0.8, and is unchanged, but abnormal rate has raise 0.78 percentage point, A: eupyrene sperm; B: teratosperm.
Embodiment
Below in conjunction with specific embodiment, the present invention is described in detail.
1 test material and method
1.1 experimental animal
Selection health is anosis, neat 30 of the large-scale New Zealand buck that block that grow up of normal 1～2 one full year of life of fertility.
1.2 test apparatus and reagent
Test apparatus: other glass apparatus such as microscope, electronic balance, PH meter, thermostat water bath, high-pressure sterilizing pot, refrigerator, aseptic operating platform, micropipette rifle, semen collector, blood counting chamber.
Test reagent: examine Ma Shi light blue dyeing liquor, Tris, citric acid, DEXTROSE ANHYDROUS (analyzing pure), yolk, penicillin, streptomycin, distilled water, 5% (percentage by weight) glucose solution, physiological saline, 3% (percentage by weight) Nacl solution etc.
1.3 semen quality measurement
1.3.1 semen collection
Adopt the artificial vagina semen collector of manual manufacture to carry out artificial semen collection.
1.3.2 ejaculatory amount
Measure with the 2.5ml syringe, carry out reading, counting behind the air in the Inside Syringe.
1.3.3 pH value is measured
Adopt the PH meter directly to measure.
1.3.4 sperm viability
By the vigor of ocular estimate with ten grades of system assessment sperms.
1.3.5 sperm concentration
The fresh semen that gathers is done 100 times of volume ratio dilutions with 3%Nacl, shake up the dilution seminal fluid, adopt blood counting chamber to count mensuration.
Number of sperm in the large grid of the number of sperm of the former seminal fluid of 1ml=5 * 5 * 10 * 1000 * 100.
1.3.6 rate of teratosperm
The dyeing of Ma Shi light blue coloring agent is examined in employing, at 1000 times of lower countings of observing.
Rate of teratosperm=teratosperm number/300 * 100%
1.4 the external preservation test of seminal fluid
1.4.1 the liquid preparation is preserved in dilution
Electronic balance accurately takes by weighing: citric acid 2.13g, and Tris3.17g, DEXTROSE ANHYDROUS 0.60g is dissolved in the 100ml distilled water, then carries out autoclaving and processes.After the complete cooling subject to sterilization, put it into aseptic operating platform, add 2ml yolk, penicillin 200,000 IU, streptomycin 200,000 IU.
1.4.2 external preservation research
Bright seminal fluid is preserved the liquid dilution with dilution, and the concentration after the dilution is 30 * 10
6/ ml.Do contrast with 5% glucose solution, physiological saline simultaneously, the contrast that is left intact of former seminal fluid.Seminal fluid after the dilution, pure seminal fluid are preserved under 4 ℃.2h after dilution, 4h, 6h, 8h, 10h, 12h, 24h, 36h, 48h, 60h, 72h carry out Observe and measure.
1.5 artificial insemination test
Adopt the autogamy dilution to preserve liquid, the seminal fluid after the dilution, vigor carries out the artificial insemination test at 0.5 time point, according to external preservation result of the test, is chosen in external preservation 60 hours point and carries out artificial insemination.Join female rabbit (blocking together large-scale New Zealand) at the beginning of selecting 500 and carry out continuously 4 tire artificial insemination, every female rabbit semen deposition concentration is 15 * 10
6The rear employing of breeding is touched the tire method and is carried out pregnancy check.
1.6 test data
After processing with Excel software, adopt the SPSS14.0 statistical software, the result all represents with mean ± standard deviation.
2 results and analysis
2.1 bright semen quality
Table 1 shows that its ejaculatory amount, vigor, pH value, density, abnormal rate are respectively 1.43ml, 0.81,7.23,3.84 * 10
8/ ml, 8.87%.
Table 1 semen quality measurement result
2.2 the external preservation result of the test of seminal fluid
Table 2 is the result show, pure seminal fluid is behind external preservation 2h, and vigor is 0.Self-control dilution is preserved the liquid group at external preservations 10h in the time, and vigor is 0.8, and is unchanged, but abnormal rate has raise 0.78 percentage point (Fig. 1), but difference not remarkable (P＞0.05); Two groups in 5% glucose solution, physiological saline, its vigor is dropping to behind the 6h below 0.5, drops to 0 behind the 10h, and abnormal rate obviously raises.Vigor, the abnormal rate significant difference of two groups of 5% glucose solutions, physiological saline and self-control dilution group each time point behind external preservation 4h, and the above two vigor significantly reduce, abnormal rate significantly raise (P＜0.05).
The external preservation of table 2 seminal fluid 10h result
Table 3 is the result show, the autogamy dilution is preserved liquid and preserved 12h outward, and sperm viability begins to descend, and when 60h, when vigor is 0.52,72h, drops to below 0.5; Abnormal rate begins significantly to raise behind external preservation 12h, is elevated to 27.11% when 72h, has raise 18.24 percentage points than former seminal fluid.
The external preservation 12～72h of table 3 seminal fluid result
2.3 artificial insemination test
Table 4 is the result show, just join, the conception rate of the 2nd tire, the 3rd tire, the 4th tire is respectively 76.40%, 75.60%, 74.00%, 74.20%, and each parity difference is not remarkable, and the population mean conception rate is 75.05%.
Table 4 artificial insemination conception rate result
3 conclusions
Result of the test shows, the sperm dilution of autogamy is preserved liquid can be more than 4 ℃ of external preservation buck seminal fluid 60h, and vigor is greater than 0.5 (vigor 0.5 is to guarantee normally to carry out test-tube index).It is higher than 5% glucose solution and physiological saline far away in external hold capacity.The conception rate of the artificial insemination test (during vigor 0.5) of carrying out simultaneously is about 75%, and result of the test shows that also this dilution prescription all can carry out artificial insemination at external preservation rabbit seminal fluid 60h with interior.
Should be understood that, for those of ordinary skills, can be improved according to the above description or conversion, and all these improvement and conversion all should belong to the protection domain of claims of the present invention.
Claims (1)
1. a rabbit sperm dilution is preserved liquid, it is characterized in that component is: citric acid 2.13g, Tris3.17g, DEXTROSE ANHYDROUS 0.60g, be dissolved in the 100ml distilled water, then carry out autoclaving and process, after the complete cooling subject to sterilization, put it into aseptic operating platform, add 2ml yolk, penicillin 200,000 IU, streptomycin 200,000 IU.
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| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN2012103612246A CN102845413A (en) | 2012-09-18 | 2012-09-18 | Diluted rabbit semen preserving fluid |
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| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN2012103612246A CN102845413A (en) | 2012-09-18 | 2012-09-18 | Diluted rabbit semen preserving fluid |
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2015027354A2 (en) | 2013-08-29 | 2015-03-05 | Universidad Austral De Chile | Method for the cooling and cryopreservation of horse semen and diluents used in said method |
| CN110622958A (en) * | 2016-06-29 | 2019-12-31 | 许红喜 | Animal sperm diluent |
Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1657020A (en) * | 2004-02-20 | 2005-08-24 | 辽宁省辽宁绒山羊原种场 | Frozen semen of animal and its making method |
| JP2007224004A (en) * | 2006-02-24 | 2007-09-06 | Yasushi Kuwabara | Diluting and preserving agent of semen for artificial insemination of ultrasmall pig |
| CN101796945A (en) * | 2010-04-12 | 2010-08-11 | 西北农林科技大学 | Antifreeze used for freezing and preserving boar semens as well as preparation and application thereof |
| CN102823580A (en) * | 2012-08-19 | 2012-12-19 | 四川省畜牧科学研究所 | Diluted preservation liquid for rabbit semen |
-
2012
- 2012-09-18 CN CN2012103612246A patent/CN102845413A/en active Pending
Patent Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1657020A (en) * | 2004-02-20 | 2005-08-24 | 辽宁省辽宁绒山羊原种场 | Frozen semen of animal and its making method |
| JP2007224004A (en) * | 2006-02-24 | 2007-09-06 | Yasushi Kuwabara | Diluting and preserving agent of semen for artificial insemination of ultrasmall pig |
| CN101796945A (en) * | 2010-04-12 | 2010-08-11 | 西北农林科技大学 | Antifreeze used for freezing and preserving boar semens as well as preparation and application thereof |
| CN102823580A (en) * | 2012-08-19 | 2012-12-19 | 四川省畜牧科学研究所 | Diluted preservation liquid for rabbit semen |
Non-Patent Citations (3)
| Title |
|---|
| 娄安钢 等: "家兔精液冷冻保存的研究", 《延边大学农学学报》, vol. 31, no. 1, 31 March 2009 (2009-03-31), pages 57 - 60 * |
| 张德福 等: "家畜精液冷冻保存技术研究进展", 《国外畜牧学-猪与禽》, vol. 25, no. 4, 31 July 2005 (2005-07-31), pages 28 - 31 * |
| 杨素芳 等: "兔精液冷冻保存的初步研究", 《广西农业生物科学》, vol. 25, 30 September 2006 (2006-09-30), pages 205 - 206 * |
Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2015027354A2 (en) | 2013-08-29 | 2015-03-05 | Universidad Austral De Chile | Method for the cooling and cryopreservation of horse semen and diluents used in said method |
| CN110622958A (en) * | 2016-06-29 | 2019-12-31 | 许红喜 | Animal sperm diluent |
| CN110622958B (en) * | 2016-06-29 | 2022-02-01 | 许红喜 | Animal sperm diluent |
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