CN106417259A - Boar seminal fluid long-acting diluting powder and application thereof - Google Patents
Boar seminal fluid long-acting diluting powder and application thereof Download PDFInfo
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- CN106417259A CN106417259A CN201610948546.9A CN201610948546A CN106417259A CN 106417259 A CN106417259 A CN 106417259A CN 201610948546 A CN201610948546 A CN 201610948546A CN 106417259 A CN106417259 A CN 106417259A
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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
- A01N1/0215—Disinfecting agents, e.g. antimicrobials for preserving living parts
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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
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Abstract
The invention belongs to the field of swine reproduction, and relates to boar seminal fluid long-acting diluting powder and an application thereof. The boar seminal fluid long-acting diluting powder is prepared from, by weight, formula 1, which comprises 4-6 g of glucose, 5-7 g of trisodium citrate dihydrate, 2-3 g of citric acid monohydrate, 7-8 g of edetic acid, 5-6 g of sodium bicarbonate, 2-3 g of potassium chloride, 6-7 g of tris(hydroxymethyl) aminomethane, 0.1-0.3 g of cysteine, 2-3 g of trehalose, 0.3-0.5 g of antibiotic enrofloxacin and 0.05-0.07 g of polymyxin B standard solution; formula 2 (optimized), which comprises 5.06 g of glucose, 6.12 g of trisodium citrate dehydrate, 2.3 g of citric acid monohydrate, 7.38 g of edetic acid, 5.54 g of sodium bicarbonate, 2.37 g of potassium chloride, 6.85 g of tris(hydroxymethyl) aminomethane, 0.21 g of cysteine, 2.11 g of trehalose, 0.42 g of antibiotic enrofloxacin and 0.06 g of polymyxin B standard solution.
Description
Technical field
The invention belongs to pig reproduction technique field.The specifically related to long-acting dilution powder of pig semen and its application.
Background technology
Artificial Insemination Technology of Swine is to gather boar semen with manual implement and seminal fluid is carried out after quality inspection and dilution, then
With apparatus, seminal fluid is injected in heat sow reproductive tract, to replace a kind of breeding method of boar natural mating, 80 years 20th century
Generation worldwide rapid popularization, at present in the use area such as wider Europe and North America, popularity rate more than 80% is
The science of carrying out pig, realize Swine Production modernization important means, it has the following advantages:(1) boar is saved, and reduces per head
The access times of boar, overcome time and area differentiation, easily can breed in good time;(2) human and material resources, financial resources are saved, carries
High economic benefit, reduces transmission of disease;(3) breeding rate is improved, due to utilizing the instrument such as microscope, Density Measuring Instrument to sperm
Quality is monitored, and effectively increases conception rate, and can adjust the poor boar of sperm quality in time;(4) increase heredity
The utilization rate of the boar of function admirable, expands breeding coverage rate, is conducive to breed improvement and union breeding.
In the artificial insemination work for carrying out pig, it is necessary to prepare the diluent of boar semen, preserve so as to dilution.
Application number 201310478237.6 discloses a kind of diluting boar semen agent, and application number 201010133910.9 is disclosed
A kind of porcine semen at normal temperature preserves diluent and its preparation and dilution process, preserves the seminal fluid time all only 3 days;Application number
201410164618.1 pig antiviral semen diluent, the preservation seminal fluid time only has 4 days.
The disclosure of the invention of the Application No. 201410527131.5 long-acting semen dilution formula of liquid of a boar antiviral and system
Preparation Method, the time for preserving seminal fluid is 5-7 days, and its defect is that polyvinyl alcohol in the preparation in formula is needed in heating electromagnetic
It is dissolved on agitator in the distilled water of 150ml 30-50 DEG C, 20-40 minute is stirred, preparation process is bothered.
A kind of long-acting diluting boar semen of the disclosure of the invention of the application number 201210238501.4 closely related with the present invention
Powder and preparation method thereof, the holding time of pig semen is 10-12 days, but dry chicken egg-yolk powder preparation method therein is complicated, system
During standby, contaminated probability is big, needs corresponding drying equipment, and quality guarantee period is short.
So develop and preserving pig semen time length, preparing easy boar semen dilution powder to technology of artificial insemination with weight
Want meaning.
Content of the invention
It is an object of the invention to overcoming the defect of prior art, a kind of long-lasting boar semen dilution powder being provided and its is answered
With, the present invention draw materials easily, prepare easy, be easy to storage, apply the present invention the Semen routine of pig long-acting semen dilution powder when
Between be 9-11 days.
It is a further object of the invention to provide a kind of application process of the long-acting dilution powder of pig semen.
Technical scheme is as follows:
A kind of long-acting dilution powder of pig semen, it is mixed by the component of following weight portion:Glucose 4-6g, two water lemons
Lemon acid trisodium 5-7g, citric acid monohydrate 2-3g, ethylenediaminetetraacetic acid 7-8g, sodium bicarbonate 5-6g, potassium chloride 2-3g, trihydroxy methyl
Aminomethane 6-7g, cysteine 0.1-0.3g, trehalose 2-3g, antibiotics enrofloxacin 0.3-0.5g and sulphuric acid polymyxin
0.05-0.07g.
Preferably, the present invention is mixed by the component of following weight portion:Glucose 5.06g, citrate dihydrate
Trisodium 6.12g, citric acid monohydrate 2.30g, ethylenediaminetetraacetic acid 7.38g, sodium bicarbonate 5.54g, potassium chloride 2.37g, three hydroxyl first
Base aminomethane 6.85g, cysteine 0.21g, trehalose 2.11g, antibiotics enrofloxacin 0.42g and sulphuric acid polymyxin
0.06g.
A kind of application process of the long-acting dilution powder of pig semen, comprises the steps:Collection boar semen, determines and is permeated
It is 7.04 ± 0.05 to press as 301.25 ± 14.52mmol/kg, pH value, according to the preparation purpose of the long-acting dilution powder of pig semen -- right
Sperm is harmless, can provide sperm nutrition, and pH scope is suitable for sperm survival, and osmotic pressure is appropriate, and raw material is easy to get, configures simply, long-acting
Preserve, the screening operation of component is carried out, screening each component for obtaining is:Glucose, citrate dihydrate trisodium
(C6H5Na307.2H2O), citric acid monohydrate, ethylenediaminetetraacetic acid (EDTA), sodium bicarbonate (NaHCo3), potassium chloride (Kcl), three
Hydroxymethyl aminomethane (TRIS), cysteine, trehalose, antibiotics enrofloxacin and sulphuric acid polymyxin, bovine serum albumin
In vain (BSA).
According to the boar semen osmotic pressure and pH value and seminar that obtain pre- formula result of the test in advance is determined, by cattle
Serum albumin, cysteine, trehalose, antibiotics enrofloxacin and sulphuric acid polymyxin are used as Main change factor, gradient
Level is 4, is tested using L16 (45) orthogonal table, and 3 are repeated (seminal fluid of 3 boars), screening formula.Then to formula
Optimize further, physical and chemical parameter is adjusted, based on osmotic pressure, osmotic pressure gradient test is carried out, by optimal osmotic pressure by formula
Optimize further, it is contemplated that bovine serum albumin is expensive, cost increase directly resulted in, be unfavorable for large-scale promotion application,
Decision is abandoned.Each test period is 11 days or so, stops inspection after most semen sample vigor are reduced to 0.6
Survey, analysis obtains optimal formula, the formula dilution seminal fluid after optimization, preserves 9-11 days, and motility of sperm is 0.60.
The invention has the beneficial effects as follows:
During natural mating, boar can undertake 25 sows every year and carry out the breeding load of this friendship, and using manually awarding
Essence, a boar can join more than 250 sows for 1 year, and artificial insemination is compared with this friendship, and its advantage is to save the feeding of boar
Support amount.The long-acting dilution powder dilution pig semen of the application present invention carries out artificial insemination, it is possible to reduce the sterilization in artificial insemination procedures
With the number of times for joining diluent, dilution seminal fluid, pig semen saves to use at any time within the time of a fixed length, energy-saving
Thing.The pig semen of the pig semen long-acting dilution powder dilution that is prepared with the present invention under the constant temperature that temperature is 17 DEG C, Ke Yibao
Deposit 9-11 days.
Specific embodiment
Below by embodiment, the present invention is described in further details, embodiment is only used for the present invention is described, does not limit
The scope of the present invention processed;Embodiment is raw materials used using the pure rank of analysis, and all steps are all carried out in better purifying space, better purifying space
Purity requirements be 300,000 grades.
Embodiment 1
A kind of long-acting dilution powder of pig semen, is mixed with the component of following weight portion:Glucose 4.80g, two water Fructus Citri Limoniaes
Sour trisodium 5.80g, citric acid monohydrate 2.18g, ethylenediaminetetraacetic acid 7.00g, sodium bicarbonate 5.25g, potassium chloride 2.25g, three hydroxyls
Aminomethane 6.50g, cysteine 0.20g, trehalose 2.00g, antibiotics enrofloxacin 0.40g and sulphuric acid polymyxin
0.06g.
(1) embodiment raw material is sealed after weighing, vacuum packaging, is obtained dilution powder, 2-8 DEG C of bar using analysis pure rank
Preserve under part.
(2), when using dilution powder, with the redistilled water stirring for being heated to 35-37 DEG C, mix homogeneously, constant volume is arrived
1000ml, obtains semen diluent, after 1 hour for pig semen dilution.
(3), after pig semen collection, abnormal smells from the patient, the normal pig semen of color are first used sperm concentration instrument detection density, then pre-
Microscopy motility of sperm under 100 power microscopes on hot 37 DEG C of slides, vigor reaches more than 0.9 grade of pig semen and calculates dilution with formula
Number:Weight × density × vigor ÷ 30=number (30 refer to the effective sperm count 3,000,000,000 of per part of seminal fluid).
(4) semen diluent and the pig semen temperature difference be no more than 1 DEG C, and makes pig by the temperature for adjusting semen diluent
The temperature of seminal fluid and semen diluent is as close as, it is impossible to reverse operating.
(5) when diluting pig semen, semen diluent is poured slowly in pig semen along chamber wall, drop should not be big, with not
Spume and be defined, be then shaken gently for or with sterilization Glass rod stirring, so that both is softly mixed;Calculate by volume, such as make 1:2-4
Times dilution for many times when, should first make 1:1-1.5 times of low power dilution, by remaining semen diluent along container after half a minute
Wall is slowly added to, and can not pour pig semen in semen diluent in turn.All dilution operations will be in room temperature 23-25 DEG C ring
Under border, complete in 30min.
(6) pig semen after dilution is slowly loaded in seminal fluid bottle or the semen bag of cleaning, with whole seminal fluid bottle or seminal fluid
Bag volume calculations, air content 6-7%, closed seminal fluid bottle or semen bag, are reduced sperm and are contacted with oxygen, the generation of Inhibit sperm
Thank, extend the holding time of seminal fluid, (also can be with clean dry pure under 23-25 DEG C of room temperature after 1 hour of lucifuge slow cooling
After the big towel of cotton wraps up 2 layers) preservation in 17 DEG C of calorstats is positioned over, the slow shake seminal fluid every 8 hours, it is to avoid sperm deposition is made
Become injures and deaths.A motility of sperm was detected every 24 hours.
(7) indoor no smoking, and female operator can not use the dense cosmetics of abnormal smells from the patient, the thermometer for contacting with seminal fluid,
Glass rod, diluted cup, 37 DEG C of preheatings of dilution water.In the transportation of pig semen, sunlight will be avoided to irradiate, the vibrations that avoid jolting,
The temperature difference is avoided to rise and fall sharply and quickly.Using special constant temperature transport case or bubble chamber, summer puts frozen water bag, and winter puts cotton towel, protects
The temperature inside the box is held for 15-20 DEG C.
Implementation result is shown in Table 1
Embodiment 2
A kind of long-acting dilution powder of the pig semen of optimization, is mixed by the component of following weight portion:Glucose 5.06g, two
Water citric acid trisodium 6.12g, citric acid monohydrate 2.30g, ethylenediaminetetraacetic acid 7.38g, sodium bicarbonate 5.54g, potassium chloride
2.37g, trishydroxymethylaminomethane 6.85g, cysteine 0.21g, trehalose 2.11g, antibiotics enrofloxacin 0.42g and sulfur
Sour polymyxin 0.06g.
(1) embodiment raw material is sealed after weighing, vacuum packaging, is obtained dilution powder, 2-8 DEG C of bar using analysis pure rank
Preserve under part.
(2), when using dilution powder, with the redistilled water stirring for being heated to 35-37 DEG C, mix homogeneously, constant volume is arrived
1000ml, obtains semen diluent, after 1 hour for pig semen dilution.
(3), after pig semen collection, abnormal smells from the patient, the normal pig semen of color are first used sperm concentration instrument detection density, then pre-
Microscopy motility of sperm under 100 power microscopes on hot 37 DEG C of slides, vigor reaches more than 0.9 grade of pig semen and calculates dilution with formula
Number:Weight × density × vigor ÷ 30=number (30 refer to the effective sperm count 3,000,000,000 of per part of seminal fluid).
(4) semen diluent and the pig semen temperature difference be no more than 1 DEG C, and makes pig by the temperature for adjusting semen diluent
The temperature of seminal fluid and semen diluent is as close as, it is impossible to reverse operating.
(5) when diluting pig semen, semen diluent is poured slowly in pig semen along chamber wall, drop should not be big, with not
Spume and be defined, be then shaken gently for or with sterilization Glass rod stirring, so that both is softly mixed;Calculate by volume, such as make 1:2-4
Times dilution for many times when, should first make 1:1-1.5 times of low power dilution, by remaining semen diluent along container after half a minute
Wall is slowly added to, and can not pour pig semen in semen diluent in turn.All dilution operations will be in room temperature 23-25 DEG C ring
Under border, complete in 30min.
(6) pig semen after dilution is slowly loaded in seminal fluid bottle or the semen bag of cleaning, with whole seminal fluid bottle or seminal fluid
Bag volume calculations, air content 6-7%, closed seminal fluid bottle or semen bag, are reduced sperm and are contacted with oxygen, the generation of Inhibit sperm
Thank, extend the holding time of seminal fluid, (also can be with clean dry pure under 23-25 DEG C of room temperature after 1 hour of lucifuge slow cooling
After the big towel of cotton wraps up 2 layers) preservation in 17 DEG C of calorstats is positioned over, the slow shake seminal fluid every 8 hours, it is to avoid sperm deposition is made
Become injures and deaths.A motility of sperm was detected every 24 hours.
(7) indoor no smoking, and female operator can not use the dense cosmetics of abnormal smells from the patient, the thermometer for contacting with seminal fluid,
Glass rod, diluted cup, 37 DEG C of preheatings of dilution water.In the transportation of pig semen, sunlight will be avoided to irradiate, the vibrations that avoid jolting,
The temperature difference is avoided to rise and fall sharply and quickly.Using special constant temperature transport case or bubble chamber, summer puts frozen water bag, and winter puts cotton towel, protects
The temperature inside the box is held for 15-20 DEG C.Test effect is shown in Table 1
1 boar semen of table preserves vigour changes statistics under 17 DEG C of constant temperature
The pig semen of the long-acting dilution powder dilution after present invention optimization, under the constant temperature that temperature is 17 DEG C, preserves 9-
11 days, motility of sperm was 0.60.Scientific investigations showed that, the sperm of straight ahead motion accounts for total sperm count between 60-95%, people
The pregnancy rate no significant difference of work insemination sow.
Claims (3)
1. the long-acting dilution powder of pig semen, it is characterised in that mixed by following components in parts by weight:Glucose 4-6g, two
Water citric acid trisodium 5-7g, citric acid monohydrate 2-3g, ethylenediaminetetraacetic acid 7-8g, sodium bicarbonate 5-6g, potassium chloride 2-3g, three hydroxyls
Aminomethane 6-7g, cysteine 0.1-0.3g, trehalose 2-3g, antibiotic enrofloxacin 0.3-0.5g and sulphuric acid are sticked more
Rhzomorph 0.05-0.07g.
2. the long-acting dilution powder of pig semen, it is characterised in that the long-acting dilution powder of described pig semen be by following components in parts by weight
Mix:Glucose 5.06g, citrate dihydrate trisodium 6.12g, citric acid monohydrate 2.30g, ethylenediaminetetraacetic acid 7.38g, carbon
Sour hydrogen sodium 5.54g, potassium chloride 2.37g, trishydroxymethylaminomethane 6.85g, cysteine 0.21g, trehalose 2.11g, antibiosis
Plain enrofloxacin 0.42g and sulphuric acid polymyxin 0.06g.
3. application of the long-acting dilution powder of pig semen described in claim 1 or 2 in pig supplementary reproduction.
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Cited By (5)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN108077244A (en) * | 2018-01-31 | 2018-05-29 | 于文龙 | A kind of long-acting boar semen dilution powder |
CN111436420A (en) * | 2020-04-14 | 2020-07-24 | 武汉市金驰海科技有限公司 | Long-acting fresh pig semen diluting preservative and using method thereof |
CN113545336A (en) * | 2021-07-19 | 2021-10-26 | 杨凌职业技术学院 | Pig semen purifying agent |
CN114208814A (en) * | 2021-12-27 | 2022-03-22 | 湖北楚钧达生物科技有限公司 | Diluent for frozen pig semen and its preparing process and application |
CN117296830A (en) * | 2023-09-25 | 2023-12-29 | 仲恺农业工程学院 | Boar semen diluent and preparation method and application thereof |
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Cited By (5)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN108077244A (en) * | 2018-01-31 | 2018-05-29 | 于文龙 | A kind of long-acting boar semen dilution powder |
CN111436420A (en) * | 2020-04-14 | 2020-07-24 | 武汉市金驰海科技有限公司 | Long-acting fresh pig semen diluting preservative and using method thereof |
CN113545336A (en) * | 2021-07-19 | 2021-10-26 | 杨凌职业技术学院 | Pig semen purifying agent |
CN114208814A (en) * | 2021-12-27 | 2022-03-22 | 湖北楚钧达生物科技有限公司 | Diluent for frozen pig semen and its preparing process and application |
CN117296830A (en) * | 2023-09-25 | 2023-12-29 | 仲恺农业工程学院 | Boar semen diluent and preparation method and application thereof |
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