CN114097769A - Pig semen dilution powder and processing technology thereof - Google Patents

Pig semen dilution powder and processing technology thereof Download PDF

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Publication number
CN114097769A
CN114097769A CN202111494237.6A CN202111494237A CN114097769A CN 114097769 A CN114097769 A CN 114097769A CN 202111494237 A CN202111494237 A CN 202111494237A CN 114097769 A CN114097769 A CN 114097769A
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Prior art keywords
percent
powder
semen
diluted
raw materials
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CN202111494237.6A
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Chinese (zh)
Inventor
孙瑶
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Shandong Daohe Biotechnology Co ltd
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Shandong Daohe Biotechnology Co ltd
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Priority to CN202111494237.6A priority Critical patent/CN114097769A/en
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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
    • 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
    • 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)
  • Agricultural Chemicals And Associated Chemicals (AREA)

Abstract

The invention discloses a pig semen diluting powder and a processing technology thereof, wherein the pig semen diluting powder is prepared by mixing the following raw materials in percentage by weight: 12-15% of trisodium citrate dihydrate, 10-13% of trihydroxymethyl aminomethane, 5-8% of anhydrous citric acid, 4-6% of ethylene diamine tetraacetic acid disodium dihydrate, 1-2% of sodium bicarbonate and 100% of anhydrous glucose. The diluted boar semen powder can prolong the in-vitro survival time of the boar semen, and can reach 13 days at the constant temperature of 17 ℃.

Description

Pig semen dilution powder and processing technology thereof
Technical Field
The invention relates to the technical field of pig breeding, in particular to pig semen diluent powder and a processing technology thereof.
Background
The artificial insemination technology for pigs is a breeding method for collecting boar semen by using an artificial instrument, inspecting and diluting the quality of the semen and then injecting the semen into the reproductive tract of an oestrous sow by using the instrument to replace natural mating of boars, is rapidly popularized in the world in the 80 th 20 th century, has the popularization rate of over 80 percent in the widely used regions such as Europe, North America and the like at present, is an important means for scientifically raising pigs and realizing the modernization of pig raising production, and has the following advantages: (1) saving boars, reducing the use frequency of each boar, overcoming time and area difference and being convenient for timely hybridization; (2) the labor, material and financial resources are saved, the economic benefit is improved, and the spread of diseases is reduced; (3) the breeding rate is improved, and the sperm quality can be monitored by using instruments such as a microscope, a densimeter and the like, so that the breeding conception rate is effectively improved, and boars with poor sperm quality can be timely adjusted; (4) increase the utilization rate of boars with excellent genetic performance, enlarge the coverage rate of improved breeds and be beneficial to breed improvement and combined breeding.
In the process of carrying out artificial insemination work of pigs, a diluent of boar semen must be prepared so as to be diluted and stored.
Chinese patent document CN103518703A discloses a boar semen diluent, which contains glucose, trisodium citrate, potassium chloride, polyvinyl alcohol, trimethylolmethane, citric acid, ciprofloxacin, glutamine dipeptide and antifreeze protein. By using the diluent, the sperm motility reaches 0.87%, the sperm acrosome integrity reaches 91.2%, and the sperm aberration rate is 7.67%, so that the sperm retention time can be effectively prolonged. But the porcine semen diluent preserves semen for only 3 days.
Chinese patent document CN101803595A discloses a diluent for preserving boar semen at normal temperature, which comprises the following components in parts by weight: 9 to 11 portions of glucose, 0.2 to 0.3 portion of ethylene diamine tetraacetic acid disodium, 0.5 to 0.6 portion of trisodium citrate, 0.08 to 0.12 portion of penicillin of 1570U/mg and 0.2 to 0.25 portion of streptomycin of 720U/mg. Completely dissolving the diluent in distilled water according to the dilution ratio that 1g of the diluent is diluted by 20ml of distilled water to obtain a diluent with the pH of 6.5-6.8 and the osmotic pressure of 290-310 mOsmo/L; the boar semen diluent produced by the formula can effectively prolong the sperm retention time. But the porcine semen diluent preserves semen for only 3 days.
Chinese patent document CN105010304A discloses a formula composition of pig antiviral semen diluent. Based on per 1000ml of double distilled water, the porcine antiviral semen diluent contains: 15.50g of glucose; 11.65g of trisodium citrate; 2.35g of EDTA sodium salt; 1.75g of sodium bicarbonate; 1.00g of polyvinyl alcohol; 5.50g of tricarboxymethylaminomethane; 4.10g of citric acid; 0.07g of cysteine; 1.00g of vitamin C; 0.10g of enrofloxacin; and 50ml of antiviral agent. The diluent composed by the formula of the porcine antiviral semen diluent provided by the embodiment of the invention has obvious killing, inhibiting and blocking effects on Porcine Reproductive and Respiratory Syndrome Virus (PRRSV), and can obviously improve the survival rate and the preservation time of sperm in vitro preservation. However, the porcine antiviral semen diluent can be stored for only 4 days.
Chinese patent document CN104322483A discloses a formula and a preparation method of a porcine antiviral long-acting semen diluent, and the porcine semen diluent comprises the following components: the preparation method comprises the steps of dissolving the components in distilled water, uniformly mixing, and sterilizing to obtain a pig semen diluent; the formula contains abundant nutrient substances, antiviral Chinese and western medicinal components and antibacterial components, can ensure normal metabolism of sperm, has effects of nutrition and preventing sperm shock after a certain time at the lower limit of preservation temperature, and can avoid invasion of external bacteria and viruses in artificial fertilization process. But the time for storing the semen by the porcine antiviral long-acting semen diluent is 5-7 days. In addition, the polyethylene ethanol in the formula needs to be dissolved in 150mL of 30-50 ℃ distilled water on a heating electromagnetic stirrer and stirred for 20-40 minutes, and the preparation steps are troublesome.
Chinese patent document CN102726371A discloses a method for preparing a long-acting pig semen diluent powder, which is prepared from glucose powder: 26-36 g, chicken yolk dry powder: 3-7 g, sodium citrate: 3-6 g, glycine: 1.5-2.5 g, disodium hydrogen phosphate: 1.0-2.0 grams, disodium edetate: 1.0-2.0 g, potassium sorbate: 2.0-4.0 g, calcium propionate: 1.0-2.0 g and 0.005-0.02 g of gentamicin powder. And uniformly mixing the powder raw materials taken out according to the amount to obtain the boar semen diluent powder. The preparation method of the chicken yolk dry powder comprises the following steps: cooking fresh ovum gallus Domesticus, taking out ovum gallus Domesticus flavus, and drying to obtain dried ovum gallus Domesticus flavus powder with water content of 7% -10%. The nutrient solution prepared by the invention and distilled water for diluting the boar semen not only contains nutrient, diluent, buffering agent, pH regulator and antibiotic, but also contains antiseptic and bacteriostatic agent, and can be preserved for 10-12 days under the constant temperature condition of 15-20 ℃. But the preparation method of the chicken yolk dry powder is complex, the pollution probability in the preparation process is high,
corresponding drying equipment is needed, and the quality guarantee period is short.
Chinese patent document CN106417259A discloses a long-acting pig semen diluent and application thereof. The composition is prepared by mixing the following components in parts by weight: formula 1: 4-6g of glucose, 5-7g of trisodium citrate dihydrate, 2-3g of citric acid monohydrate, 7-8g of ethylenediamine tetraacetic acid, 5-6g of sodium bicarbonate, 2-3g of potassium chloride, 6-7g of tris (hydroxymethyl) aminomethane, 0.1-0.3g of cysteine, 2-3g of trehalose, 0.3-0.5g of enrofloxacin antibiotic and 0.05-0.07g of polymyxin sulfate; formulation 2 (optimized): 5.06g of glucose, 6.12g of trisodium citrate dihydrate, 2.30g of citric acid monohydrate, 7.38g of ethylenediamine tetraacetic acid, 5.54g of sodium bicarbonate, 2.37g of potassium chloride, 6.85g of tris (hydroxymethyl) aminomethane, 0.21g of cysteine, 2.11g of trehalose, 0.42g of the antibiotic enrofloxacin and 0.06g of polymyxin sulfate. The boar semen diluted by the prepared boar semen long-acting dilution powder can be stored for 9-11 days under the constant temperature condition of 17 ℃. However, the invention uses cysteine, trehalose, antibiotics enrofloxacin and polymyxin sulfate, and the temperature is constant at 17 ℃ for up to 11 days.
Disclosure of Invention
In view of the above, the present invention aims to provide a diluted pig semen powder and a processing method thereof, wherein the diluted pig semen powder can prolong the in vitro survival time of pig semen, and can last up to 13 days at a constant temperature of 17 ℃.
The adopted technical scheme is as follows:
the invention relates to a pig semen diluent powder, which is prepared by mixing the following raw materials in percentage by weight:
12 to 15 percent of trisodium citrate dihydrate,
10 to 13 percent of trihydroxymethyl aminomethane,
5 to 8 percent of anhydrous citric acid,
4 to 6 percent of ethylene diamine tetraacetic acid disodium dihydrate,
1 to 2 percent of sodium bicarbonate,
anhydrous glucose was added to 100%.
Further, the pig semen diluent powder is prepared by mixing the following raw materials in percentage by weight:
14.79 percent of trisodium citrate dihydrate,
12.11 percent of trihydroxymethyl aminomethane,
6.22 percent of anhydrous citric acid,
5.04 percent of ethylene diamine tetraacetic acid disodium dihydrate,
1.50 percent of sodium bicarbonate,
anhydrous glucose was added to 100%.
Further, 47 plus or minus 1g of the pig semen diluent powder is added into 1000mL of deionized water for complete dissolution, and the pH value is measured to be 6.6-7.0.
Furthermore, 47 +/-1 g of the diluted porcine semen powder is taken and added into 1000mL of deionized water for complete dissolution, and the osmotic pressure is determined to be 280-310 mOsm.
The invention relates to a processing technology of pig semen diluted powder, which comprises the following steps:
s1, cleaning and fully drying an HGD-2000 mixer in advance, keeping the indoor humidity below 60 percent,
s2, smashing trihydroxymethyl aminomethane and raw materials with particle size larger than 40 meshes; then weighing the weight of each raw material according to the formula;
s3, adding the raw materials into an HGD-2000 mixer, wherein the mixing time is 20-30min, and the rotating speed is 10-20r/min;
and S4, after subpackaging, sampling and detecting.
The invention has the beneficial effects that:
the pig semen diluent powder supplements nutrient substances for sperms, maintains the osmotic pressure of the sperms, reduces the damage of bacteria in the sperms to the sperms, reduces the damage of sperm metabolites to the sperms, reduces the damage of active oxygen in the sperms to the sperm plasma membrane structure, effectively protects the integrity of the sperm plasma membrane structure and acrosome integrity, can effectively prolong the survival time of the pig sperms stored in vitro, and can reach up to 13 days under the constant temperature condition of 17 ℃. Thereby improving the semen use efficiency of the high-quality breeding boars and the economic benefit of artificial breeding.
Detailed Description
The present invention is described in detail below with reference to specific examples, but the use and purpose of these exemplary embodiments are merely to exemplify the present invention, and do not set forth any limitation on the actual scope of the present invention in any form, and the scope of the present invention is not limited thereto.
Example 1
The pig semen diluent powder is prepared by mixing the following raw materials in percentage by weight:
12 percent of trisodium citrate dihydrate,
10 percent of trihydroxymethyl aminomethane,
5 percent of anhydrous citric acid,
4 percent of ethylene diamine tetraacetic acid disodium dihydrate,
1 percent of sodium bicarbonate,
anhydrous glucose was added to 100%.
The processing technology of the pig semen diluent powder comprises the following steps:
s1, cleaning and fully drying an HGD-2000 mixer in advance, keeping the indoor humidity below 60 percent,
s2, smashing trihydroxymethyl aminomethane and raw materials with particle size larger than 40 meshes; then weighing the weight of each raw material according to the formula;
s3, adding the raw materials into an HGD-2000 mixer, wherein the mixing time is 20min, and the rotating speed is 12r/min;
and S4, after subpackaging, sampling and detecting, releasing sale if the quality is qualified, and destroying if the quality is not qualified.
[ product storage ]: shading, sealing, and storing in shade at ambient temperature not higher than 30 deg.C.
[ packaging specifications ]: 47 plus or minus 1 g/bag; 188 + -4 g/bag.
The product quality inspection process comprises the following steps:
【1】 Particle size
Taking 1 bag (47 + -1 g) of the product, and checking with 40 mesh sieve until the granules larger than 40 mesh are no more than 10%;
【2】 Clarity of solution
Adding 1 bag (47 + -1 g) of the product into 1000ml of distilled water, dissolving, and the dissolved liquid is colorless and clear and contains no visible foreign matter;
【3】 pH value
Taking 1 bag (47 +/-1 g) of the product, adding the product into 1000ml of distilled water, and measuring according to the pH value measuring method operating procedure to be 6.6-7.0;
【4】 Osmotic pressure
Taking 1 bag (47 +/-1 g) of the product, adding 1000ml of distilled water, and measuring according to the osmometry operating procedure, wherein the concentration is 280-310 mOSM.
Example 2
Referring to the embodiment 1, different from the embodiment 1, the diluted pig semen powder of the embodiment is prepared by mixing the following raw materials in percentage by weight:
14.79 percent of trisodium citrate dihydrate,
12.11 percent of trihydroxymethyl aminomethane,
6.22 percent of anhydrous citric acid,
5.04 percent of ethylene diamine tetraacetic acid disodium dihydrate,
1.50 percent of sodium bicarbonate,
anhydrous glucose was added to 100%.
The rest is the same as example 1.
Example 3
Referring to the embodiment 1, different from the embodiment 1, the diluted pig semen powder of the embodiment is prepared by mixing the following raw materials in percentage by weight:
15 percent of trisodium citrate dihydrate,
13 percent of trihydroxymethyl aminomethane,
8 percent of anhydrous citric acid,
6 percent of ethylene diamine tetraacetic acid disodium dihydrate,
2 percent of sodium bicarbonate,
anhydrous glucose was added to 100%.
The rest is the same as example 1.
Test and test:
the diluted powder of the pig semen of the embodiment 2 is put into 1000mL of double distilled water, and is continuously stirred by a magnetic stirrer until the diluted powder is completely dissolved, and the diluted solution is prepared by heating the diluted powder to 35 ℃. According to the pig semen: volume ratio of the dilution liquid = 1: 5, diluting, and slowly adding a diluent during diluting. And stored at a constant temperature of 17 deg.C, sperm motility was observed every 24 hours, and motility changes were recorded (10 replicates). The results are shown in Table 1:
TABLE 1 Change in storage Activity at constant temperature of 17 ℃
Retention time Example 2
2h 92.43±0.41
24h 91.52±0.23
48h 90.36±0.14
72h 89.21±0.35
96h 88.10±0.22
120h 86.63±0.27
144h 84.72±0.18
168h 82.12±0.72
192h 80.14±0.85
216h 78.63±0.79
240h 76.62±0.51
264h 74.01±0.12
288h 72.87±0.32
312h 70.22±0.18
As can be seen from Table 1, the sperm motility of example 2 is still above 70% after storing for 312 hours (13 days), and the diluted boar semen powder prolongs the in vitro survival time of the boar semen, and can reach up to 13 days under the constant temperature condition of 17 ℃.
The above-listed detailed description is only a specific description of a possible embodiment of the present invention, and they are not intended to limit the scope of the present invention, and equivalent embodiments or modifications made without departing from the technical spirit of the present invention should be included in the scope of the present invention.

Claims (5)

1. The boar semen diluent powder is characterized by being prepared by mixing the following raw materials in percentage by weight:
12 to 15 percent of trisodium citrate dihydrate,
10 to 13 percent of trihydroxymethyl aminomethane,
5 to 8 percent of anhydrous citric acid,
4 to 6 percent of ethylene diamine tetraacetic acid disodium dihydrate,
1 to 2 percent of sodium bicarbonate,
anhydrous glucose was added to 100%.
2. The boar semen diluent powder according to claim 1, which is prepared by mixing the following raw materials in percentage by weight:
14.79 percent of trisodium citrate dihydrate,
12.11 percent of trihydroxymethyl aminomethane,
6.22 percent of anhydrous citric acid,
5.04 percent of ethylene diamine tetraacetic acid disodium dihydrate,
1.50 percent of sodium bicarbonate,
anhydrous glucose was added to 100%.
3. The diluted boar semen powder as recited in claim 1, wherein 47 ± 1g of the diluted boar semen powder is taken and added into 1000mL of deionized water for complete dissolution, and the measured pH value is 6.6-7.0.
4. The diluted boar semen powder as recited in claim 1, wherein 47 ± 1g of the diluted boar semen powder is taken and added into 1000mL of deionized water for complete dissolution, and the measured osmotic pressure is 280-310 mOsm.
5. A process for the preparation of a diluted powder of porcine semen according to any of claims 1 to 4, characterized in that it comprises the following steps:
s1, cleaning and fully drying an HGD-2000 mixer in advance, keeping the indoor humidity below 60 percent,
s2, smashing trihydroxymethyl aminomethane and raw materials with particle size larger than 40 meshes; then weighing the weight of each raw material according to the formula;
s3, adding the raw materials into an HGD-2000 mixer, wherein the mixing time is 20-30min, and the rotating speed is 10-20r/min;
and S4, after subpackaging, sampling and detecting.
CN202111494237.6A 2021-12-08 2021-12-08 Pig semen dilution powder and processing technology thereof Pending CN114097769A (en)

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Application Number Priority Date Filing Date Title
CN202111494237.6A CN114097769A (en) 2021-12-08 2021-12-08 Pig semen dilution powder and processing technology thereof

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Citations (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
JP2007224004A (en) * 2006-02-24 2007-09-06 Yasushi Kuwabara Diluting and preserving agent of semen for artificial insemination of ultrasmall pig
WO2010005260A2 (en) * 2008-07-10 2010-01-14 주식회사 노아바이오텍 Diluent for long-term preservation of liquid boar sperm
CN102687715A (en) * 2011-03-23 2012-09-26 北京浩邦猪人工授精服务有限责任公司 Cold-water instant pig seminal fluid dilution powder formula and manufacturing process and application method
CN104068012A (en) * 2014-06-17 2014-10-01 宜川欣源生态养殖专业合作社 Formula of cold water instant type pig seminal fluid diluting powder as well as production process and application method

Patent Citations (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
JP2007224004A (en) * 2006-02-24 2007-09-06 Yasushi Kuwabara Diluting and preserving agent of semen for artificial insemination of ultrasmall pig
WO2010005260A2 (en) * 2008-07-10 2010-01-14 주식회사 노아바이오텍 Diluent for long-term preservation of liquid boar sperm
CN102687715A (en) * 2011-03-23 2012-09-26 北京浩邦猪人工授精服务有限责任公司 Cold-water instant pig seminal fluid dilution powder formula and manufacturing process and application method
CN104068012A (en) * 2014-06-17 2014-10-01 宜川欣源生态养殖专业合作社 Formula of cold water instant type pig seminal fluid diluting powder as well as production process and application method

Non-Patent Citations (3)

* Cited by examiner, † Cited by third party
Title
K.F.WEITZE: "猪精液常温保存液", 《国外畜牧科技》 *
赵书广: "《中国养猪大成》", 31 March 2001, 中国农业出版社 *
鲜红等: "猪精液常温保存研究进展", 《黑龙江动物繁殖》 *

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