CN105660609A - Bovine seminal fluid preservation method - Google Patents
Bovine seminal fluid preservation method Download PDFInfo
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- CN105660609A CN105660609A CN201610254576.XA CN201610254576A CN105660609A CN 105660609 A CN105660609 A CN 105660609A CN 201610254576 A CN201610254576 A CN 201610254576A CN 105660609 A CN105660609 A CN 105660609A
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- seminal fluid
- diluent
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- dilution
- semen
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- 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
- A01N1/0205—Chemical aspects
- A01N1/021—Preservation or perfusion media, liquids, solids or gases used in the preservation of cells, tissue, organs or bodily fluids
- A01N1/0221—Freeze-process protecting agents, i.e. substances protecting cells from effects of the physical process, e.g. cryoprotectants, osmolarity regulators like oncotic agents
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- 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)
- Micro-Organisms Or Cultivation Processes Thereof (AREA)
Abstract
The invention discloses a bovine seminal fluid preservation method which comprises the following steps: (1) seminal fluid collection; (2) diluent preparation; (3) seminal fluid dilution; (4) seminal fluid freezing. According to the bovine seminal fluid preservation method, requirements on conditions are low; the operation is easy to implement; the bovine seminal fluid preservation method is suitable for large-scale breeding and reproduction.
Description
Technical field
The present invention relates to biological technical field, it is specifically related to a kind of ox Semen routine method.
Background technology
It is that twentieth century is produced along with technology of artificial insemination widespread use that livestock semen preserves technology, and principle is by reducing or Inhibit sperm metabolism degree reaches the object extending sperm life-span in vitro. For this reason, investigator's test preserves under seminal fluid is placed on low-temperature condition (non-icing) or freezing (freezing) state. But in temperature changing process, the protection ability of sperm is limited by seminal fluid. For this reason, carry out semen cryopreservation first to be diluted by seminal fluid with the use of special solution. The basic demand of diluent is comprised pH value, resiliency, osmotic pressure and low-temperature protection performance.
Summary of the invention
The technical problem to be solved in the present invention is to provide a kind of ox Semen routine method.
Solving the problems of the technologies described above, the technical solution used in the present invention is:
A kind of ox Semen routine method, comprises the following steps:
(1) seminal fluid collection: gather the ox seminal fluid that ripening degree is good, for subsequent use under depositing in 6-7 DEG C of clean environment;
(2) diluent preparing: take sterile purified water as solvent, every 1000ml diluent consists of the following composition: glycerine 5-10 gram, soybean lecithin 1-2 gram, trehalose 8-12 gram, vitamin-E 2-4 gram, 10-15 gram, yolk, trisodium citrate 4-7 gram, penicillin 80-100 ten thousand IU, all the other be solvent, regulate PH value be 6.6-7.0;
(3) semen dilution: with the diluent of isothermal, the dense part ox seminal fluid adopted doubly is done first time dilution by 1:0.5 at 30 DEG C, be cooled to 15 DEG C through 1 hour; Do second time dilution with the diluent of isothermal by former essence amount 1:1, it is cooled to 5 DEG C through 23 hours; Third time dilution is done by former essence amount 1:1 again with the diluent of isothermal;
(4) semen freezing: seminal fluid is loaded tubule, is put in refrigerated cylinder.
The average surviving rate of ox seminal fluid of described collection is greater than 80%;
The seminal fluid sampling of described step (3) after dilution mirror inspection rate alive at 30-40 DEG C is not less than former seminal fluid.
It is an advantage of the invention that:
1, the diluent formula of step (2) is reasonable, effectively extends Sperm survival time;
2, adopting freezing semen method to preserve sperm, cooling rate is accelerated, it is possible to protection sperm damages from low temperature in cooling, refrigerating process;
3, condition of the present invention requires low, and operation is easy to realize, and is suitable for large-scale cultivation and breeds.
Embodiment
Below embodiments of the invention are described in detail, so that advantages and features of the invention can be easier to be readily appreciated by one skilled in the art, thus protection scope of the present invention are made more explicit defining.
Embodiment 1
A kind of ox Semen routine method, comprises the following steps:
(1) seminal fluid collection: gathering the ox seminal fluid that ripening degree is good, the average surviving rate of ox seminal fluid of collection is greater than 80%, for subsequent use under depositing in 6-7 DEG C of clean environment;
(2) diluent preparing: take sterile purified water as solvent, every 1000ml diluent consists of the following composition: glycerine 5 grams, soybean lecithin 1 gram, trehalose 8 grams, vitamin-E 2 grams, 10 grams, yolk, trisodium citrate 4 grams, penicillin 800,000 IU, all the other be solvent, regulate PH value be 6.6;
(3) semen dilution: with the diluent of isothermal, the dense part ox seminal fluid adopted doubly is done first time dilution by 1:0.5 at 30 DEG C, be cooled to 15 DEG C through 1 hour; Do second time dilution with the diluent of isothermal by former essence amount 1:1, it is cooled to 5 DEG C through 23 hours; Doing dilution for the third time with the diluent of isothermal by former essence amount 1:1 again, the mirror inspection rate alive at 30-40 DEG C of the seminal fluid sampling after dilution is not less than former seminal fluid;
(4) semen freezing: seminal fluid is loaded tubule, is put in refrigerated cylinder.
Embodiment 2
A kind of ox Semen routine method, comprises the following steps:
(1) seminal fluid collection: gathering the ox seminal fluid that ripening degree is good, the average surviving rate of ox seminal fluid of collection is greater than 80%, for subsequent use under depositing in 6-7 DEG C of clean environment;
(2) diluent preparing: take sterile purified water as solvent, every 1000ml diluent consists of the following composition: glycerine 7.5 grams, soybean lecithin 1.5 grams, trehalose 10 grams, vitamin-E 3 grams, 12.5 grams, yolk, trisodium citrate 5.5 grams, penicillin 900,000 IU, all the other be solvent, regulate PH value be 6.8;
(3) semen dilution: with the diluent of isothermal, the dense part ox seminal fluid adopted doubly is done first time dilution by 1:0.5 at 30 DEG C, be cooled to 15 DEG C through 1 hour; Do second time dilution with the diluent of isothermal by former essence amount 1:1, it is cooled to 5 DEG C through 23 hours; Doing dilution for the third time with the diluent of isothermal by former essence amount 1:1 again, the mirror inspection rate alive at 30-40 DEG C of the seminal fluid sampling after dilution is not less than former seminal fluid;
(4) semen freezing: seminal fluid is loaded tubule, is put in refrigerated cylinder.
Embodiment 3
A kind of ox Semen routine method, comprises the following steps:
(1) seminal fluid collection: gathering the ox seminal fluid that ripening degree is good, the average surviving rate of ox seminal fluid of collection is greater than 80%, for subsequent use under depositing in 6-7 DEG C of clean environment;
(2) diluent preparing: take sterile purified water as solvent, every 1000ml diluent consists of the following composition: glycerine 10 grams, soybean lecithin 2 grams, trehalose 12 grams, vitamin-E 4 grams, 15 grams, yolk, trisodium citrate 7 grams, penicillin 1,000,000 IU, all the other be solvent, regulate PH value be 7.0;
(3) semen dilution: with the diluent of isothermal, the dense part ox seminal fluid adopted doubly is done first time dilution by 1:0.5 at 30 DEG C, be cooled to 15 DEG C through 1 hour; Do second time dilution with the diluent of isothermal by former essence amount 1:1, it is cooled to 5 DEG C through 23 hours; Doing dilution for the third time with the diluent of isothermal by former essence amount 1:1 again, the mirror inspection rate alive at 30-40 DEG C of the seminal fluid sampling after dilution is not less than former seminal fluid;
(4) semen freezing: seminal fluid is loaded tubule, is put in refrigerated cylinder.
Claims (3)
1. an ox Semen routine method, it is characterised in that comprise the following steps:
(1) seminal fluid collection: gather the ox seminal fluid that ripening degree is good, for subsequent use under depositing in 6-7 DEG C of clean environment;
(2) diluent preparing: take sterile purified water as solvent, every 1000ml diluent consists of the following composition: glycerine 5-10 gram, soybean lecithin 1-2 gram, trehalose 8-12 gram, vitamin-E 2-4 gram, 10-15 gram, yolk, trisodium citrate 4-7 gram, penicillin 80-100 ten thousand IU, all the other be solvent, regulate PH value be 6.6-7.0;
(3) semen dilution: with the diluent of isothermal, the dense part ox seminal fluid adopted doubly is done first time dilution by 1:0.5 at 30 DEG C, be cooled to 15 DEG C through 1 hour; Do second time dilution with the diluent of isothermal by former essence amount 1:1, it is cooled to 5 DEG C through 23 hours; Third time dilution is done by former essence amount 1:1 again with the diluent of isothermal;
(4) semen freezing: seminal fluid is loaded tubule, is put in refrigerated cylinder.
2. ox Semen routine method according to claim 1, it is characterised in that the average surviving rate of ox seminal fluid of described collection is greater than 80%.
3. according to the ox Semen routine method described in claim 1 or 2, it is characterised in that the seminal fluid sampling of described step (3) after dilution mirror inspection rate alive at 30-40 DEG C is not less than former seminal fluid.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201610254576.XA CN105660609A (en) | 2016-04-22 | 2016-04-22 | Bovine seminal fluid preservation method |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201610254576.XA CN105660609A (en) | 2016-04-22 | 2016-04-22 | Bovine seminal fluid preservation method |
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| CN105660609A true CN105660609A (en) | 2016-06-15 |
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| Application Number | Title | Priority Date | Filing Date |
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| CN201610254576.XA Pending CN105660609A (en) | 2016-04-22 | 2016-04-22 | Bovine seminal fluid preservation method |
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Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN107333752A (en) * | 2017-08-08 | 2017-11-10 | 重庆信首科技有限公司 | Kind ox seminal fluid Cryopreservation |
| CN109221085A (en) * | 2018-09-25 | 2019-01-18 | 上海祥欣畜禽有限公司 | The fresh essence of pig saves dilution powder |
| CN111053079A (en) * | 2016-06-29 | 2020-04-24 | 许红喜 | Preparation method of animal sperm diluent |
Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2001037655A1 (en) * | 1999-11-24 | 2001-05-31 | Xy, Inc. | Method of cryopreserving selected sperm cells |
| CN1729770A (en) * | 2005-08-11 | 2006-02-08 | 西北农林科技大学 | Chinese herbal medicine diluent for frozen bovine semen |
| CN101220345A (en) * | 2008-01-23 | 2008-07-16 | 西北农林科技大学 | Cattle freezing seminal fluid dilution and method for producing the same |
| CN101310729A (en) * | 2007-05-23 | 2008-11-26 | 辽宁省辽宁绒山羊育种中心 | Animal origin free flocks and herds frozen semen diluent and production method of flocks and herds frozen semen |
-
2016
- 2016-04-22 CN CN201610254576.XA patent/CN105660609A/en active Pending
Patent Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2001037655A1 (en) * | 1999-11-24 | 2001-05-31 | Xy, Inc. | Method of cryopreserving selected sperm cells |
| CN1729770A (en) * | 2005-08-11 | 2006-02-08 | 西北农林科技大学 | Chinese herbal medicine diluent for frozen bovine semen |
| CN101310729A (en) * | 2007-05-23 | 2008-11-26 | 辽宁省辽宁绒山羊育种中心 | Animal origin free flocks and herds frozen semen diluent and production method of flocks and herds frozen semen |
| CN101220345A (en) * | 2008-01-23 | 2008-07-16 | 西北农林科技大学 | Cattle freezing seminal fluid dilution and method for producing the same |
Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN111053079A (en) * | 2016-06-29 | 2020-04-24 | 许红喜 | Preparation method of animal sperm diluent |
| CN111053079B (en) * | 2016-06-29 | 2022-02-01 | 许红喜 | Preparation method of animal sperm diluent |
| CN107333752A (en) * | 2017-08-08 | 2017-11-10 | 重庆信首科技有限公司 | Kind ox seminal fluid Cryopreservation |
| CN109221085A (en) * | 2018-09-25 | 2019-01-18 | 上海祥欣畜禽有限公司 | The fresh essence of pig saves dilution powder |
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Application publication date: 20160615 |
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