CN107333752A - Kind ox seminal fluid Cryopreservation - Google Patents
Kind ox seminal fluid Cryopreservation Download PDFInfo
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- CN107333752A CN107333752A CN201710669949.4A CN201710669949A CN107333752A CN 107333752 A CN107333752 A CN 107333752A CN 201710669949 A CN201710669949 A CN 201710669949A CN 107333752 A CN107333752 A CN 107333752A
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- Prior art keywords
- seminal fluid
- dilution
- time
- cryopreservation
- preheating
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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
- A01N1/0226—Physiologically active agents, i.e. substances affecting physiological processes of cells and tissue to be preserved, e.g. anti-oxidants or nutrients
-
- 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)
- Biophysics (AREA)
- Physiology (AREA)
- Micro-Organisms Or Cultivation Processes Thereof (AREA)
Abstract
The invention discloses kind of an ox seminal fluid Cryopreservation, comprise the following steps：The collection of seminal fluid, the detection of seminal fluid, the preparation of dilution, semen dilution, freezen protective.Present invention kind carries out the dilution of seminal fluid so as to reaching target viscosities in three times, the temperature of the ox seminal fluid after dilution can be reduced after often once being diluted, the survival rate of ox seminal fluid at low temperature is high, slowly reduce the storage temperature of seminal fluid, be advantageous to improve sperm motility rate, the enough Acclimation temperature time is provided for sperm.
Description
Technical field
The present invention relates to a kind of Semen routine method, and in particular to kind ox seminal fluid Cryopreservation.
Background technology
The freezen protective of seminal fluid plays a very important role in Modern Animal Husbandry, using frozen semen can not by when
Between, region and the limitation in breeding stock life-span, reduce male animal feeding quantity, reduce production cost, improve the utilization rate of excellent poultry kind, be
Animal husbandry production brings huge economic benefit.Meanwhile the freezing of seminal fluid and the use of frozen semen are raiseeing kind of introduction, blood relationship more
Newly, there is extremely important meaning on disease control, endangered breed and genetic diversity conservation.
That To Be Protected from Heat is not cold for ox seminal fluid, at present, in order to reduce the viscosity of ox seminal fluid, dilution can be used to carry out ox seminal fluid dilute
Release, typically can more than 30 DEG C at a temperature of to seminal fluid carry out once completely dilution, seminal fluid more than 30 DEG C at a temperature of survive
Time is short, still, in order to ensure that dilution and ox seminal fluid are thoroughly mixed and reach aimed concn, it is necessary to longer dilution time,
So seminal fluid more than 30 degrees Celsius at a temperature of the holdup time grow, the survival rate of seminal fluid is low.
The content of the invention
The technical problems to be solved by the invention be ox seminal fluid under Cord blood, how to ensure higher sperm survival
Rate solves the problems, such as survival rate of the ox seminal fluid under Cord blood, and it is an object of the present invention to provide kind ox seminal fluid Cryopreservation.
The present invention is achieved through the following technical solutions：
Kind ox seminal fluid Cryopreservation, comprises the following steps：
S1, seminal fluid collection：The good ox seminal fluid of maturity is gathered, is deposited in standby under 2-4 DEG C of sterile environment；
S2, seminal fluid detection：Quality Detection is carried out to the seminal fluid in step S1；
S3, dilution preparation：Prepare the dilution with frozen semen function；
S4, semen dilution：The seminal fluid up to standard to Quality Detection in step S2 is carried out dilute after dilution preheating in step S3
Release, the dilution step includes following sub-step：
A, after the dilution in step S3 is preheating to 30-37 DEG C, by dilution: the volume ratio of seminal fluid is 1: 0.5 pair of seminal fluid
First time dilution is carried out, seminal fluid is cooled to 15-20 DEG C and balanced 1 hour after diluting for the first time；
B, after the dilution in step S3 is preheating to 30-37 DEG C, by dilution: the volume ratio of seminal fluid is in 1: 1 couple of step A
The seminal fluid after dilution carries out second of dilution for the first time, is cooled to 5-10 DEG C after second of dilution of seminal fluid and balances 1 hour, described
The dilution liquid measure of second of dilution is based on the semen volume before first time dilution；
C, after the dilution in step S3 is preheating to 30-37 DEG C, by dilution: the volume ratio of seminal fluid is in 1: 1 couple of step B
Seminal fluid after second of dilution carries out third time dilution, is cooled to 2-4 DEG C after second of dilution of seminal fluid and balances 1 hour, described
The dilution liquid measure of third time dilution is based on the semen volume before first time dilution；
S5, freezen protective：Seminal fluid in step S4 after diluting three times is freezed with sequencing frigorimeter, freezing knot
Inspect by random samples and moved into liquid nitrogen container after beam and preserved and used.
The dilution of the invention for carrying out seminal fluid in three times can reduce dilute so as to reach target viscosities, after often once being diluted
The temperature of ox seminal fluid after releasing, the survival rate of ox seminal fluid at low temperature is high, slowly reduces the storage temperature of seminal fluid, is advantageous to improve
Sperm motility rate, the enough Acclimation temperature time is provided for sperm；Cord blood mainly suppresses sperm motility using low temperature, drop
Low metabolism and energy expenditure, are suppressed growth of microorganism, while add necessary nutrition and other compositions, and completely cut off air, to reach
Extend the purpose of Sperm survival time；The present invention is excellent first to have selected the preferable seminal fluid of quality, ensure that the seminal fluid preserved
With stronger viability, so as to be advantageous in semen dilution, the survival rate of sperm in seminal fluid is improved.
Quality Detection includes following detection in step S2：Seminal fluid density, sperm motility rate, seminal fluid acid-base property, sperm survival
Time.
Dilution includes following components in the step S3：Ultra-pure water, glycerine, soybean lecithin, trehalose, lactose, dimension
Raw plain E, yolk, trisodium citrate, trishydroxymethylaminomethane, penicillin.Must the addition anti-cold shock of such as yolk in dilution
Material, improve sperm chilling resistance, it is also necessary to provide necessary nutriment for seminal fluid.
Dilution is preheating to 35 DEG C in described step A, B, C.It can either ensure that seminal fluid is rapidly diluted at 35 DEG C, again
Seminal fluid can be slowed down to die rapidly.
The sperm motility rate minimum standard value that step S2 is detected is 80%.Existing seminal fluid motility rate is generally 70%, this hair
It is bright that the higher seminal fluid of motility rate is selected under limited detection means, be advantageous to maintain sperm in the seminal fluid under Cord blood to survive
Rate.
The present invention compared with prior art, has the following advantages and advantages：
1st, the dilution that present invention kind ox seminal fluid Cryopreservation carries out seminal fluid in three times is often entered so as to reach target viscosities
Row can reduce the temperature of the ox seminal fluid after dilution after once diluting, the survival rate of ox seminal fluid at low temperature is high, slowly reduces essence
The storage temperature of liquid, be advantageous to improve sperm motility rate, the enough Acclimation temperature time is provided for sperm；
2nd, ox seminal fluid Cryopreservation is excellent first have selected the preferable seminal fluid of quality for present invention kind, ensure that and is preserved
Seminal fluid there is stronger viability, so as to be advantageous in semen dilution, improve seminal fluid in sperm survival rate；
3rd, present invention kind ox seminal fluid Cryopreservation step is simple, and practicality is high.
Embodiment
For the object, technical solutions and advantages of the present invention are more clearly understood, with reference to embodiment, the present invention is made
Further to describe in detail, exemplary embodiment of the invention and its explanation are only used for explaining the present invention, are not intended as to this
The restriction of invention.
Embodiment 1
Present invention kind ox seminal fluid Cryopreservation, comprises the following steps：
S1, seminal fluid collection：The good ox seminal fluid of maturity is gathered, is deposited in standby under 2-4 DEG C of sterile environment；
S2, seminal fluid detection：Quality Detection is carried out to the seminal fluid in step S1；
S3, dilution preparation：Prepare the dilution with frozen semen function；
S4, semen dilution：The seminal fluid up to standard to Quality Detection in step S2 is carried out dilute after dilution preheating in step S3
Release, the dilution step includes following sub-step：
A, after the dilution in step S3 is preheating to 30-37 DEG C, by dilution: the volume ratio of seminal fluid is 1: 0.5 pair of seminal fluid
First time dilution is carried out, seminal fluid is cooled to 15-20 DEG C and balanced 1 hour after diluting for the first time；
B, after the dilution in step S3 is preheating to 30-37 DEG C, by dilution: the volume ratio of seminal fluid is in 1: 1 couple of step A
The seminal fluid after dilution carries out second of dilution for the first time, is cooled to 5-10 DEG C after second of dilution of seminal fluid and balances 1 hour, described
The dilution liquid measure of second of dilution is based on the semen volume before first time dilution；
C, after the dilution in step S3 is preheating to 30-37 DEG C, by dilution: the volume ratio of seminal fluid is in 1: 1 couple of step B
Seminal fluid after second of dilution carries out third time dilution, is cooled to 2-4 DEG C after second of dilution of seminal fluid and balances 1 hour, described
The dilution liquid measure of third time dilution is based on the semen volume before first time dilution；
S5, freezen protective：Seminal fluid in step S4 after diluting three times is freezed with sequencing frigorimeter, freezing knot
Inspect by random samples and moved into liquid nitrogen container after beam and preserved and used.
Dilution is preheating to 35 DEG C in described step A, B, C.It can either ensure that seminal fluid is rapidly diluted at 35 DEG C, again
Seminal fluid can be slowed down to die rapidly.
The sperm motility rate minimum standard value that step S2 is detected is 80%.Existing seminal fluid motility rate is generally 70%, this hair
It is bright that the higher seminal fluid of motility rate is selected under limited detection means, be advantageous to maintain sperm in the seminal fluid under Cord blood to survive
Rate.
The dilution of the invention for carrying out seminal fluid in three times can reduce dilute so as to reach target viscosities, after often once being diluted
The temperature of ox seminal fluid after releasing, the survival rate of ox seminal fluid at low temperature is high, slowly reduces the storage temperature of seminal fluid, is advantageous to improve
Sperm motility rate, the enough Acclimation temperature time is provided for sperm；Cord blood mainly suppresses sperm motility using low temperature, drop
Low metabolism and energy expenditure, are suppressed growth of microorganism, while add necessary nutrition and other compositions, and completely cut off air, to reach
Extend the purpose of Sperm survival time；The present invention is excellent first to have selected the preferable seminal fluid of quality, ensure that the seminal fluid preserved
With stronger viability, so as to be advantageous in semen dilution, the survival rate of sperm in seminal fluid is improved.
Quality Detection includes following detection in step S2：Seminal fluid density, sperm motility rate, seminal fluid acid-base property, sperm survival
Time.
Embodiment 2
Based on embodiment 1, dilution includes following components in the step S3：Ultra-pure water, glycerine, soybean lecithin, sea
Algae sugar, lactose, vitamin E, yolk, trisodium citrate, trishydroxymethylaminomethane, penicillin.Must addition such as ovum in dilution
The anti-cold shock material such as Huang, improve sperm chilling resistance, it is also necessary to provide necessary nutriment for seminal fluid.
Above-described embodiment, the purpose of the present invention, technical scheme and beneficial effect are carried out further
Describe in detail, should be understood that the embodiment that the foregoing is only the present invention, be not intended to limit the present invention
Protection domain, within the spirit and principles of the invention, any modification, equivalent substitution and improvements done etc., all should include
Within protection scope of the present invention.
Claims (5)
1. kind of ox seminal fluid Cryopreservation, it is characterised in that comprise the following steps：
S1, seminal fluid collection：The good ox seminal fluid of maturity is gathered, is deposited in standby under 2-4 DEG C of sterile environment；
S2, seminal fluid detection：Quality Detection is carried out to the seminal fluid in step S1；
S3, dilution preparation：Prepare the dilution with frozen semen function；
S4, semen dilution：The seminal fluid up to standard to Quality Detection in step S2 is diluted after dilution preheating in step S3, institute
Stating dilution step includes following sub-step：
A, after the dilution in step S3 is preheating to 30-37 DEG C, by dilution: the volume ratio of seminal fluid is that 1: 0.5 pair of seminal fluid is carried out
Dilute for the first time, seminal fluid is cooled to 15-20 DEG C and balanced 1 hour after diluting for the first time；
B, after the dilution in step S3 is preheating to 30-37 DEG C, by dilution: the volume ratio of seminal fluid is first in 1: 1 couple of step A
Seminal fluid after secondary dilution carries out second and diluted, and seminal fluid is cooled to 5-10 DEG C after diluting for second and balances 1 hour, and described second
The dilution liquid measure of secondary dilution is based on the semen volume before first time dilution；
C, after the dilution in step S3 is preheating to 30-37 DEG C, by dilution: the volume ratio of seminal fluid is second in 1: 1 couple of step B
Seminal fluid after secondary dilution carries out third time dilution, and seminal fluid is cooled to 2-4 DEG C after diluting for second and balances 1 hour, and the described 3rd
The dilution liquid measure of secondary dilution is based on the semen volume before first time dilution；
S5, freezen protective：Seminal fluid in step S4 after diluting three times is freezed with sequencing frigorimeter, after freezing terminates
Inspect by random samples and move into liquid nitrogen container and preserved and used.
2. according to claim a kind of ox seminal fluid Cryopreservation, it is characterised in that Quality Detection includes in step S2
Following detection：Seminal fluid density, sperm motility rate, seminal fluid acid-base property, Sperm survival time.
3. according to claim a kind of ox seminal fluid Cryopreservation, it is characterised in that dilution bag in the step S3
Include following components：Ultra-pure water, glycerine, soybean lecithin, trehalose, lactose, vitamin E, yolk, trisodium citrate, three hydroxyl first
Base aminomethane, penicillin.
4. according to claim a kind of ox seminal fluid Cryopreservation, it is characterised in that diluted in described step A, B, C
Liquid is preheating to 35 DEG C.
5. according to claim 2 kind of ox seminal fluid Cryopreservation, it is characterised in that the sperm that step S2 is detected is lived
Rate minimum standard value is 80%.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201710669949.4A CN107333752A (en) | 2017-08-08 | 2017-08-08 | Kind ox seminal fluid Cryopreservation |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201710669949.4A CN107333752A (en) | 2017-08-08 | 2017-08-08 | Kind ox seminal fluid Cryopreservation |
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| Publication Number | Publication Date |
|---|---|
| CN107333752A true CN107333752A (en) | 2017-11-10 |
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ID=60216797
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201710669949.4A Pending CN107333752A (en) | 2017-08-08 | 2017-08-08 | Kind ox seminal fluid Cryopreservation |
Country Status (1)
| Country | Link |
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| CN (1) | CN107333752A (en) |
Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| 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 |
| CN111345283A (en) * | 2020-03-31 | 2020-06-30 | 上海市农业科学院 | Method for freezing and storing buffalo semen |
| CN112586494A (en) * | 2020-12-18 | 2021-04-02 | 洛阳市洛瑞牧业有限公司 | Fried smoking method frozen beef sperm manufacturing process |
Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103704203A (en) * | 2014-01-03 | 2014-04-09 | 甘肃省畜牧兽医研究所 | Preparation method and using method for GL and TCM199 serving as bovine semen diluent and bovine semen refrigerating fluid |
| WO2016041033A1 (en) * | 2014-09-15 | 2016-03-24 | Perez Eduardo Gualtieri De Andrade | Method for thawing and refreezing cryopreserved semen |
| CN105660609A (en) * | 2016-04-22 | 2016-06-15 | 李志新 | Bovine seminal fluid preservation method |
-
2017
- 2017-08-08 CN CN201710669949.4A patent/CN107333752A/en active Pending
Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103704203A (en) * | 2014-01-03 | 2014-04-09 | 甘肃省畜牧兽医研究所 | Preparation method and using method for GL and TCM199 serving as bovine semen diluent and bovine semen refrigerating fluid |
| WO2016041033A1 (en) * | 2014-09-15 | 2016-03-24 | Perez Eduardo Gualtieri De Andrade | Method for thawing and refreezing cryopreserved semen |
| CN105660609A (en) * | 2016-04-22 | 2016-06-15 | 李志新 | Bovine seminal fluid preservation method |
Non-Patent Citations (2)
| Title |
|---|
| BAILEY J ET AL: "Semen cryopreservation:Successes and persistent problems in farm species", 《CANADIAN JOURNAL OF ANIMAL SCIENCE》 * |
| 赵占强等: "不同浓度海藻糖对牛冷冻精液品质的影响", 《中国牛业科学》 * |
Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| 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 |
| 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 |
| CN112586494A (en) * | 2020-12-18 | 2021-04-02 | 洛阳市洛瑞牧业有限公司 | Fried smoking method frozen beef sperm manufacturing process |
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Application publication date: 20171110 |
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