CN106993605A - A kind of down producing goat freezing seminal fluid dilution formula - Google Patents
A kind of down producing goat freezing seminal fluid dilution formula Download PDFInfo
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- CN106993605A CN106993605A CN201710180689.4A CN201710180689A CN106993605A CN 106993605 A CN106993605 A CN 106993605A CN 201710180689 A CN201710180689 A CN 201710180689A CN 106993605 A CN106993605 A CN 106993605A
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- down producing
- seminal fluid
- producing goat
- yolk
- fluid dilution
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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
-
- 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
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- Health & Medical Sciences (AREA)
- Engineering & Computer Science (AREA)
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- General Health & Medical Sciences (AREA)
- Wood Science & Technology (AREA)
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- Micro-Organisms Or Cultivation Processes Thereof (AREA)
Abstract
A kind of down producing goat freezing seminal fluid dilution formula, the formula is made up of 2.7~3.3g of glucose, 5.1~5.7g of lactose, 1.1~1.4g of sodium citrate, yolk 7~12%, heat lowlenthal serum 17~23%, ethylene glycol 4~6% and above-mentioned substance is dissolved into 100ml pure water, and above-mentioned percentage is percent by volume；The present invention also provides a kind of down producing goat freezing seminal fluid dilution compound method.The present invention promotes sperm ovum binding, so as to improve conception rate by add a certain amount of heat lowlenthal serum in down producing goat Oocytes in Vitro Fertilization nutrient solution and suitably reduce yolk content.
Description
Technical field：
The present invention relates to down producing goat reproduction technique field, more particularly to a kind of down producing goat freezing seminal fluid dilution formula.
Background technology：
In recent years, goat artificial insemination obtains more and more universal application on livestock culturing, especially for scale,
Intensive, specialized cultivation, technology of artificial insemination has played important function.Semen cryopreservation technology ensure that artificial insemination pair
The demand of seminal fluid is not limited by time and region.Freezen protective technology causes semen collection and fertilization to generate over time and space
Separation, extends the external storage time of sperm, is easy to long-distance transport, promotes trans-regional, the transnational exchange of genetic resources
With it is shared, accelerate drove breed improvement paces so that outstanding poultry kind is promoted on a large scale.
The freezen protective of sperm is a complicated process, satisfactorily result is frozen to obtain, in refrigerating process
In need balance many factors.Need suitable diluent, antifreeze and other additives, equilibrium temperature and equilibration time, drop
Warm speed and Thawing Rate are all vital, and its each single item is likely to influence semen quality.In semen cryopreservation,
Cryoprotector is the key that sperm super-low temperature freezes storage success or not.Its main function is to reduce cell in process of freezing and thawing
Permeability is damaged and reduced ice crystal and formed, and cell is able to preserve under condition of ultralow temperature.
At present, cryoprotector is broadly divided into permeability cryoprotector (such as glycerine, ethylene glycol) and impermeability is cold
Freeze protective agent (such as monose, yolk, albumin).It is generally acknowledged that yolk is due to that rich in lecithin, can improve extracellular osmotic pressure,
Stable spermatoblast film, prevents outer acrosomal membrane rupturing, recent research is also demonstrated, under the conditions of non-frozen, yolk can promote
Sperm is combined with egg vitellary membrane, so as to improve acrosome reaction rate.Existed yet between yolk and bulbourethral gland secretion
Harmful interaction, can decline living spermatozoa percentage after rewarming.
Research of the China to goat sperm freezen protective technology is started from the 1980s, starting to walk evening.To 80 years 20th century
For the middle and later periods, begin attempt to be applied to production practices.Goat sperm conception rate after freeze thawing is relatively low, is usually no more than 50%, in fact
Border productive value is relatively low, the bottleneck as restriction goat sperm freezing.Therefore, we are necessary to match somebody with somebody the dilution of frozen semen
Fang Jinhang is studied and improved.
The content of the invention：
In consideration of it, being necessary to design a kind of down producing goat freezing seminal fluid dilution formula to improve the conception rate that goat freezes essence；
Meanwhile, also to provide a kind of down producing goat freezing seminal fluid dilution compound method.
A kind of down producing goat freezing seminal fluid dilution formula, the formula by 2.7~3.3g of glucose, 5.1~5.7g of lactose,
1.1~1.4g of sodium citrate, yolk 7~12%, heat lowlenthal serum 17~23%, ethylene glycol 4~6% and by above-mentioned substance
100ml pure water composition is dissolved to, above-mentioned percentage is percent by volume.
It is preferred that, glucose 3g, lactose 5.5g, sodium citrate 1.3g, yolk 10%, heat lowlenthal serum 20%, second two
Alcohol 5%.
A kind of down producing goat freezing seminal fluid dilution compound method, comprises the following steps：
Step 1: weighing 2.7~3.3g of glucose, 5.1~5.7g of lactose, 1.1~1.4g of sodium citrate, fully it is dissolved into
In 50ml pure water, this liquid is solution I；
Step 2: take one piece of Fresh Egg, yolk is suctioned out with syringe, and with the filtering of 45 micron sterile filters, take 7~
12ml adds solution I；
Step 3: down producing goat concentrates during oestrus, centrifugal process prepares heat lowlenthal serum, takes 17~23ml to add solution I；
Step 4: taking 4~6ml of ethylene glycol to add solution I, and above-mentioned substance is dissolved to 100ml with pure water；
Step 5: with 22 micron sterile filter filtration sterilizations, and 4 DEG C of preservations are stand-by, now with the current, storage life is no more than 7
My god.
It is preferred that, glucose 3g, lactose 5.5g, sodium citrate 1.3g, yolk 10%, heat lowlenthal serum 20%, second two
Alcohol 5%.
Heat lowlenthal serum is cells with nutrient and growth factor in cell cultivation process, to maintaining cytoactive to have
Important function.The present invention in down producing goat Oocytes in Vitro Fertilization nutrient solution by that will add a certain amount of heat lowlenthal serum
And yolk content is suitably reduced, promote sperm ovum binding, so as to improve conception rate.
Embodiment：
First, the preparation of down producing goat freezing seminal fluid dilution：
1st, glucose 3g, lactose 5.5g are weighed, sodium citrate 1.3g, is fully dissolved into 50ml pure water, this liquid is solution
I；
2nd, one piece of Fresh Egg is taken, yolk is suctioned out with syringe, and is filtered with 45 micron sterile filters, takes 10ml to add molten
Liquid I；
3rd, down producing goat concentrates during oestrus, and centrifugal process prepares heat lowlenthal serum, takes 20ml to add solution I；
4th, take ethylene glycol 5ml to add solution I, and above-mentioned substance is dissolved to 100ml with pure water；
5th, with 22 micron sterile filter filtration sterilizations, and 4 DEG C of preservations are stand-by, now with the current, and storage life is no more than 7 days.
2nd, seminal fluid prepares：
1st, take down producing goat stud ram seminal fluid 1ml to add the down producing goat freezing seminal fluid dilution of the foregoing preparations of 9ml, mix,
800g is centrifuged 5 minutes, removes supernatant；
2nd, the down producing goat freezing seminal fluid dilution of the foregoing preparations of 1ml is added in Sperm pellets, 10 minutes is stood, treats on sperm
After floating, supernatant (containing sperm living) tubulature or drop plate are taken, it is ensured that often manage or often drop contains the sperm of more than 1 × 107.
3rd, prepared by frozen spermatic fluid：
The present embodiment uses frozen spermatic fluid method frozen sperm.By the ready seminal fluid of previous step, 4 DEG C balance 3 hours,
The centimeters of liquid nitrogen surface 1.5 drip plate every sky, and liquid nitrogen is put into after 5 minutes.
4th, freeze smart particle to thaw：
Take out and freeze after smart particle from liquid nitrogen, be put into the sterile tube of 40 DEG C of preheatings and thaw 1 minute.
5th, defrosting semen quality is detected：
1st, sperm motility rate is determined：Sperm motility rate is determined with sperm motility detection kit.
2nd, the detection of sperm survival index：The seminal fluid of defrosting is preserved in 37 DEG C of water-baths, 1 vigor was carried out every 1 hour
Detection, until 4 hours.Sperm survival index be average and interval (hour) time of 2 inspection sperm motilities product it
With.
3rd, the detection of Sperm acrasomal integvity：Freeze after smart particle thaws and make smear, fixed through formalin phosphate liquid,
After the dyeing of Ji's nurse Sa dye liquor, in the complete and damage situations of 1000 times of oily Microscopic observation perforatoriums, the complete type essence of statistics acrosome
Sub- rate.
6th, the result of the present embodiment：
The Quality Detection result of the goat frozen spermatic fluid made of the dilution formula of liquid of the present embodiment see the table below.
| Sperm motility rate (%) | Survival index | Acrosomal integrity (%) |
| 64.12±0.57 | 6.67 | 53.24±0.46 |
7th, frozen semen artificial insemination is tested：
It is real that the jelly essence made of inventive formulation carries out artificial insemination to 100 normal Aerbasi Cashmere Goats 's of heat
Test, be 79% through counting artificial insemination conception rate.As can be seen here, the frozen semen that the formula makes has preferable practical application
Effect.
Claims (4)
1. a kind of down producing goat freezing seminal fluid dilution formula, it is characterised in that the formula is by 2.7~3.3g of glucose, lactose
5.1~5.7g, 1.1~1.4g of sodium citrate, yolk 7~12%, heat lowlenthal serum 17~23%, ethylene glycol 4~6% and
Above-mentioned substance is dissolved to 100ml pure water composition, above-mentioned percentage is percent by volume.
2. a kind of down producing goat freezing seminal fluid dilution formula as claimed in claim 1, it is characterised in that glucose 3g, lactose
5.5g, sodium citrate 1.3g, yolk 10%, heat lowlenthal serum 20%, ethylene glycol 5%.
3. a kind of down producing goat freezing seminal fluid dilution compound method, it is characterised in that comprise the following steps：
Step 1: weighing 2.7~3.3g of glucose, 5.1~5.7g of lactose, 1.1~1.4g of sodium citrate, 50ml is fully dissolved into
In pure water, this liquid is solution I；
Step 2: taking one piece of Fresh Egg, yolk is suctioned out with syringe, and is filtered with 45 micron sterile filters, takes 7~12ml to add
Enter solution I；
Step 3: down producing goat concentrates during oestrus, centrifugal process prepares heat lowlenthal serum, takes 17~23ml to add solution I；
Step 4: taking 4~6ml of ethylene glycol to add solution I, and above-mentioned substance is dissolved to 100ml with pure water；
Step 5: with 22 micron sterile filter filtration sterilizations, and 4 DEG C of preservations are stand-by, now with the current, storage life is no more than 7 days.
4. a kind of down producing goat freezing seminal fluid dilution compound method as claimed in claim 2, it is characterised in that glucose 3g,
Lactose 5.5g, sodium citrate 1.3g, yolk 10%, heat lowlenthal serum 20%, ethylene glycol 5%.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201710180689.4A CN106993605A (en) | 2017-03-17 | 2017-03-17 | A kind of down producing goat freezing seminal fluid dilution formula |
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| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201710180689.4A CN106993605A (en) | 2017-03-17 | 2017-03-17 | A kind of down producing goat freezing seminal fluid dilution formula |
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| CN106993605A true CN106993605A (en) | 2017-08-01 |
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| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201710180689.4A Pending CN106993605A (en) | 2017-03-17 | 2017-03-17 | A kind of down producing goat freezing seminal fluid dilution formula |
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN114027293A (en) * | 2021-11-26 | 2022-02-11 | 内蒙古自治区农牧业科学院 | Simple freezing method for fine tube frozen semen of cashmere goat |
| CN115251046A (en) * | 2022-08-18 | 2022-11-01 | 西安市畜牧技术推广中心 | Semen freezing diluent and application thereof in freezing preservation of milk goat semen |
-
2017
- 2017-03-17 CN CN201710180689.4A patent/CN106993605A/en active Pending
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN114027293A (en) * | 2021-11-26 | 2022-02-11 | 内蒙古自治区农牧业科学院 | Simple freezing method for fine tube frozen semen of cashmere goat |
| CN115251046A (en) * | 2022-08-18 | 2022-11-01 | 西安市畜牧技术推广中心 | Semen freezing diluent and application thereof in freezing preservation of milk goat semen |
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