CN113940340A - Chicken sperm diluent and preparation method and application thereof - Google Patents
Chicken sperm diluent and preparation method and application thereof Download PDFInfo
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- CN113940340A CN113940340A CN202111263564.0A CN202111263564A CN113940340A CN 113940340 A CN113940340 A CN 113940340A CN 202111263564 A CN202111263564 A CN 202111263564A CN 113940340 A CN113940340 A CN 113940340A
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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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- 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
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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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Abstract
The invention relates to the technical field of poultry breeding, and particularly discloses a chicken sperm diluent which is an aqueous solution and comprises the following components in mass concentration: 7-9 g/L of sodium chloride, 3.5-4 g/L of monopotassium phosphate, 13.5-15 g/L of sodium citrate dihydrate, 14-16 g/L of glucose, 0.01-0.03 g/L of curcumin and 0.03-0.05 g/L of mannitol. The chicken sperm diluent can provide nutrient substances for chicken sperm, maintain the balance of osmotic pressure inside and outside the sperm, keep the sperm activity and improve the fertilization rate; has oxidation resistance, can prolong the survival time of sperms, is beneficial to the short-term preservation and the transportation of the chicken semen at different places, and improves the utilization rate of the chicken semen.
Description
Technical Field
The invention belongs to the technical field of poultry breeding, and particularly relates to a chicken sperm diluent as well as a preparation method and application thereof.
Background
With the development of modern poultry industry, the scale and intensification degree of poultry breeding are higher and higher, and in order to improve the fertility rate, accelerate the process of fine breedings, improve the utilization rate of excellent breeding hens, reduce the feeding amount of breeding cocks and the like, the artificial insemination technology of the breeding hens becomes the most main breeding mode in modern breeding and production. The chicken semen is a highly concentrated sperm solution, has small volume and large density, directly performs artificial insemination on undiluted chicken semen, and has low fertilization rate because the undiluted chicken semen is volatile and inactive; secondly, resources are wasted, and the artificial insemination cost is increased. The chicken essence liquid is diluted, so that the volume of the chicken essence liquid can be enlarged, the number of inseminated hens is increased, the sperm activity can be maintained, the fertilization capability is improved, the semen of excellent breeders can be utilized to the maximum extent, the feeding cost can be saved, the economic benefit can be improved, and the chicken essence liquid has a leading effect on the breeding of chicken flocks. The sperm diluent provides energy for the sperm, keeps acid-base balance and maintains the balance of the osmotic pressure inside and outside the sperm. Therefore, the quality of the sperm diluent is directly related to the sperm motility, indirectly influences the fertilization rate of the poultry eggs, and finally concerns the economic benefit of the farm.
Patent No. cn201410621015.x provides a chicken semen diluent, which consists of magnesium chloride hexahydrate, potassium citrate, sodium acetate trihydrate, fructose, sodium glutamate, multivitamins, cerium chloride and a solvent, improves the sperm motility and obviously improves the utilization rate of breeder cocks, but the preservation time is short, the sperm motility is reduced to 10% after being preserved for 3 days, the sperm motility is low and the aberration rate is high after being preserved, and the sperm produced by cocks can be utilized and scientifically researched in a cross-time, cross-region, multiple modes and the like. Sperm are highly sensitive to reactive oxygen species because their plasma membranes are rich in polyunsaturated fatty acids, which are the main targets of oxidation, and their cytoplasm is largely confined to the midgut, with little antioxidant mechanism to provide adequate protection against oxidative damage, which in turn can be associated with loss of viability, low fertilization rates, or embryo developmental disorders, which is one of the major causes of reduced sperm motility during semen preservation. Therefore, the chicken essence diluent is developed, the sperm activity is maintained, the oxidative damage of the sperm is reduced, the preservation time of the chicken essence is prolonged, and the preservation and transportation of the chicken sperm are facilitated, so that the utilization rate of the chicken essence and the use value in the field of practical production and scientific research are improved.
Disclosure of Invention
The invention aims to provide a chicken sperm diluent, a preparation method and application thereof, which can improve the vitality of artificial insemination chicken sperm and prolong the preservation time of the chicken sperm.
In order to achieve the purpose, the invention provides a chicken sperm diluent, which is an aqueous solution and comprises the following components in mass concentration: 7-9 g/L of sodium chloride, 3.5-4 g/L of monopotassium phosphate, 13.5-15 g/L of sodium citrate dihydrate, 14-16 g/L of glucose, 0.01-0.03 g/L of curcumin and 0.03-0.05 g/L of mannitol.
Preferably, in the chicken sperm dilution, the chicken sperm dilution comprises the following components by mass: 8g/L of sodium chloride, 3.8g/L of monopotassium phosphate, 14g/L of sodium citrate dihydrate, 15g/L of glucose, 0.02g/L of curcumin and 0.04g/L of mannitol.
Preferably, the diluted chicken essence is stored in a refrigerator at 4 ℃ for later use.
The preparation method of the chicken essence diluent comprises the following steps: under the aseptic condition, adding 7-9 g of sodium chloride, 3.5-4 g of monopotassium phosphate, 13.5-15 g of sodium citrate dihydrate and 14-16 g of glucose into 900ml of sterilized distilled water, fully dissolving and uniformly mixing, adjusting the pH to 7.5-8, adding 0.01-0.03 g of curcumin and 0.03-0.05 g of mannitol, uniformly mixing, and then adding sterilized distilled water to a constant volume of 1L to obtain the chicken sperm diluent.
The application of the chicken sperm diluent comprises the steps of preheating the chicken sperm diluent to 36-38 ℃ in a water bath, mixing and diluting collected fresh chicken sperm and the sperm diluent according to the volume ratio of 1:1, shaking up gently and fully to obtain diluted chicken sperm, and carrying out artificial insemination or storage on the diluted chicken sperm.
Preferably, in the application of the chicken sperm diluent, the breeding hens are subjected to artificial insemination, and the input amount of the diluted chicken sperm is 0.02-0.03 ml.
Preferably, in the application of the chicken sperm diluent, the diluted chicken sperm is placed into a freezing storage tube, and is placed at 10 ℃ for 0.5-1 h, and then is placed at 4 ℃ for storage. The sperms are placed at 10 ℃ for a period of time and then placed at 4 ℃ for storage, so that the temperature of the sperms is relatively slowly reduced, and the sperms can be prevented from being stimulated by severe temperature change to generate stress effect to cause sperm inactivation.
Preferably, in the application of the chicken sperm diluent, the preserved diluted chicken sperm liquid is put into a water bath to be preheated to 36-38 ℃, and then is used for artificial insemination of the hens.
Compared with the prior art, the invention has the following beneficial effects:
1. the chicken sperm diluent can provide nutrient substances for chicken sperm, maintain the balance of osmotic pressure inside and outside the sperm, keep the sperm activity and improve the fertilization rate; has oxidation resistance, can prolong the survival time of sperms, is beneficial to short-term preservation and remote transportation of chicken semen, is beneficial to application in cross-time, cross-region and the like, and improves the utilization rate of the chicken semen.
2. According to the chicken sperm diluent, curcumin and mannitol have an anti-oxidation effect, so that oxidative stress damage of chicken sperm in the preservation process is reduced, and the preservation quality of the chicken sperm is improved; curcumin and mannitol are mutually cooperated, and the physiological activity of chicken sperms is effectively maintained in the preservation process of chicken semen by regulating and controlling the concentrations of the curcumin and the mannitol in a diluent, so that the preservation time of the chicken semen is prolonged.
3. The chicken essence diluent disclosed by the invention is few in components, simple to prepare, easy to obtain raw materials, low in cost, convenient to use, easy to popularize and suitable for large-scale poultry farms.
Drawings
Fig. 1 shows fertilization rates of 10 batches of breeding hens in application example 1 of the present invention after artificial insemination.
Detailed Description
The following detailed description of specific embodiments of the invention is provided, but it should be understood that the scope of the invention is not limited to the specific embodiments.
Example 1
A chicken sperm diluent is an aqueous solution and comprises the following components in mass concentration: 8g/L of sodium chloride, 3.8g/L of monopotassium phosphate, 14g/L of sodium citrate dihydrate, 15g/L of glucose, 0.02g/L of curcumin and 0.04g/L of mannitol.
The embodiment also provides a preparation method of the chicken sperm diluent, which comprises the following steps: under the aseptic condition, adding 8g of sodium chloride, 3.8g of monopotassium phosphate, 14g of sodium citrate dihydrate and 15g of glucose into 900ml of sterilized distilled water, fully dissolving and uniformly mixing, adjusting the pH to 7.5-8, adding 0.02g of curcumin and 0.04g of mannitol, uniformly mixing, adding sterilized distilled water, and fixing the volume to 1L to obtain the chicken sperm diluent.
Example 2
A chicken sperm diluent is an aqueous solution and comprises the following components in mass concentration: 8g/L of sodium chloride, 3.8g/L of monopotassium phosphate, 14g/L of sodium citrate dihydrate, 15g/L of glucose, 0.02g/L of curcumin and 0.05g/L of mannitol.
The procedure and parameters for the preparation of the chicken sperm diluent of this example were the same as in example 1, except that the contents of the components were different.
Example 3
A chicken sperm diluent is an aqueous solution and comprises the following components in mass concentration: 8g/L of sodium chloride, 3.8g/L of monopotassium phosphate, 14g/L of sodium citrate dihydrate, 15g/L of glucose, 0.02g/L of curcumin and 0.03g/L of mannitol.
The procedure and parameters for the preparation of the chicken sperm diluent of this example were the same as in example 1, except that the contents of the components were different.
Example 4
A chicken sperm diluent is an aqueous solution and comprises the following components in mass concentration: 8g/L of sodium chloride, 3.8g/L of monopotassium phosphate, 14g/L of sodium citrate dihydrate, 15g/L of glucose, 0.01g/L of curcumin and 0.05g/L of mannitol.
The procedure and parameters for the preparation of the chicken sperm diluent of this example were the same as in example 1, except that the contents of the components were different.
Example 5
A chicken sperm diluent is an aqueous solution and comprises the following components in mass concentration: 8g/L of sodium chloride, 3.8g/L of monopotassium phosphate, 14g/L of sodium citrate dihydrate, 15g/L of glucose, 0.03g/L of curcumin and 0.03g/L of mannitol.
The procedure and parameters for the preparation of the chicken sperm diluent of this example were the same as in example 1, except that the contents of the components were different.
Comparative example 1
A chicken sperm diluent is an aqueous solution and comprises the following components in mass concentration: 8g/L of sodium chloride, 3.8g/L of monopotassium phosphate, 14g/L of sodium citrate dihydrate and 15g/L of glucose.
The procedure and parameters of the preparation of the chicken sperm diluent of this comparative example were the same as those of example 1, except that the contents of the respective components were different.
Comparative example 2
A chicken sperm diluent is an aqueous solution and comprises the following components in mass concentration: 8g/L of sodium chloride, 3.8g/L of potassium dihydrogen phosphate, 14g/L of sodium citrate dihydrate, 15g/L of glucose and 0.06g/L of curcumin.
The procedure and parameters of the preparation of the chicken sperm diluent of this comparative example were the same as those of example 1, except that the contents of the respective components were different.
Comparative example 3
A chicken sperm diluent is an aqueous solution and comprises the following components in mass concentration: 8g/L of sodium chloride, 3.8g/L of potassium dihydrogen phosphate, 14g/L of sodium citrate dihydrate, 15g/L of glucose and 0.06g/L of mannitol.
The procedure and parameters of the preparation of the chicken sperm diluent of this comparative example were the same as those of example 1, except that the contents of the respective components were different.
Application example 1 for artificial insemination of breeding hens
Preparing the chicken sperm diluent in the example 1, preheating the chicken sperm diluent in a water bath to 37 ℃, mixing the collected fresh semen with the sperm diluent in the example 1 according to a ratio of 1:1, shaking up gently and fully, then carrying out artificial insemination on breeding hens, carrying out artificial insemination on 8000 breeding hens in 10 batches, adopting a cloaca eversion insemination method, wherein the input amount of the diluted chicken sperm is 0.025ml, and investigating the fertilization rate of eggs after insemination.
Fig. 1 shows the fertilization rates of 10 batches of breeding hens after artificial insemination, and it can be seen from the figure that the fertilization rates of 10 batches are all higher than 93%, which indicates that the sperm activity and fertilization rate after dilution are high.
Application example 2 Chicken essence liquid preserving and survival
Respectively preparing the chicken sperm diluent of the examples 1-5 and the comparative examples 1-3, preheating the chicken sperm diluent to 37 ℃ in a water bath, putting the collected fresh chicken sperm into a freezing storage tube, slowly adding the chicken sperm diluent with the volume ratio of the chicken sperm to the sperm diluent being 1:1, gently shaking and uniformly mixing to obtain the diluted chicken sperm, placing the freezing storage tube at 10 ℃ for 0.5h, and then storing at 4 ℃. Sperm motility was checked every 24 h. Specific results are shown in table 1.
TABLE 1 Chicken sperm motility Rate of diluted Chicken sperm
As can be seen from Table 1, the sperm survival rate of the example group is significantly higher than that of the comparative example group, the sperm motility rate of the example group is more than 60% after the example group is stored at low temperature for 4 days, and the sperm motility rate of the example 1 group is the highest in the whole storage process and is significantly higher than that of the group without curcumin and mannitol and the group with curcumin or mannitol alone.
The foregoing descriptions of specific exemplary embodiments of the present invention have been presented for purposes of illustration and description. It is not intended to limit the invention to the precise form disclosed, and obviously many modifications and variations are possible in light of the above teaching. The exemplary embodiments were chosen and described in order to explain certain principles of the invention and its practical application to enable one skilled in the art to make and use various exemplary embodiments of the invention and various alternatives and modifications as are suited to the particular use contemplated. It is intended that the scope of the invention be defined by the claims and their equivalents.
Claims (8)
1. The chicken sperm diluent is an aqueous solution, and comprises the following components in mass concentration: 7-9 g/L of sodium chloride, 3.5-4 g/L of monopotassium phosphate, 13.5-15 g/L of sodium citrate dihydrate, 14-16 g/L of glucose, 0.01-0.03 g/L of curcumin and 0.03-0.05 g/L of mannitol.
2. The chicken sperm dilution of claim 1, comprising the following components in mass concentration: 8g/L of sodium chloride, 3.8g/L of monopotassium phosphate, 14g/L of sodium citrate dihydrate, 15g/L of glucose, 0.02g/L of curcumin and 0.04g/L of mannitol.
3. The chicken essence dilution according to claim 1, wherein the chicken essence dilution is stored in a 4 ℃ refrigerator for later use.
4. A method of preparing a dilution of chicken essence according to claim 1 or 2, comprising the steps of: under the aseptic condition, adding 7-9 g of sodium chloride, 3.5-4 g of monopotassium phosphate, 13.5-15 g of sodium citrate dihydrate and 14-16 g of glucose into 900ml of sterilized distilled water, fully dissolving and uniformly mixing, adjusting the pH to 7.5-8, adding 0.01-0.03 g of curcumin and 0.03-0.05 g of mannitol, uniformly mixing, and then adding sterilized distilled water to a constant volume of 1L to obtain the chicken sperm diluent.
5. The application of the chicken essence diluent as claimed in claim 1, wherein the chicken essence diluent is preheated to 36-38 ℃ in water bath, collected fresh chicken essence and the sperm diluent are mixed and diluted according to the volume ratio of 1:1, the mixture is shaken up gently to obtain diluted chicken essence, and the diluted chicken essence is subjected to artificial insemination or preservation of breeding hens.
6. The use of the chicken essence diluent according to claim 1, wherein the diluted chicken essence is used for artificial insemination, and the input amount of the diluted chicken essence is 0.02-0.03 ml.
7. The use of a dilution of chicken essence according to claim 6, wherein the diluted chicken essence is preserved: placing the diluted chicken essence liquid into a freezing storage tube, placing the tube at 10 ℃ for 0.5-1 h, and then storing the tube at 4 ℃.
8. The application of the chicken essence diluent according to claim 7, wherein the preserved diluted chicken essence is placed in a water bath to be preheated to 36-38 ℃ and then used for artificial insemination of breeding hens.
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Citations (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102812950A (en) * | 2012-09-07 | 2012-12-12 | 江苏省家禽科学研究所 | Chicken semen diluent as well as preparation method and application thereof |
| CN104396941A (en) * | 2014-11-07 | 2015-03-11 | 张英芹 | Chicken sperm diluting liquid and preparation method thereof |
| KR20150101498A (en) * | 2014-02-26 | 2015-09-04 | 가천대학교 산학협력단 | Composition for cryopreservating boer sperm comprising astaxanthin or curcumin |
| CN108026544A (en) * | 2015-07-17 | 2018-05-11 | 医疗法人圣光医疗财团 | The storage method of NT cells and storage system |
| WO2019174560A1 (en) * | 2018-03-14 | 2019-09-19 | 必康生技（香港）有限公司 | Composition for purification of biofluids |
| CN111567516A (en) * | 2020-06-29 | 2020-08-25 | 扬州大学 | Semen diluent for improving fertilization rate of hatching eggs in later period of egg laying and application thereof |
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- 2021-10-28 CN CN202111263564.0A patent/CN113940340B/en active Active
Patent Citations (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102812950A (en) * | 2012-09-07 | 2012-12-12 | 江苏省家禽科学研究所 | Chicken semen diluent as well as preparation method and application thereof |
| KR20150101498A (en) * | 2014-02-26 | 2015-09-04 | 가천대학교 산학협력단 | Composition for cryopreservating boer sperm comprising astaxanthin or curcumin |
| CN104396941A (en) * | 2014-11-07 | 2015-03-11 | 张英芹 | Chicken sperm diluting liquid and preparation method thereof |
| CN108026544A (en) * | 2015-07-17 | 2018-05-11 | 医疗法人圣光医疗财团 | The storage method of NT cells and storage system |
| WO2019174560A1 (en) * | 2018-03-14 | 2019-09-19 | 必康生技（香港）有限公司 | Composition for purification of biofluids |
| CN111567516A (en) * | 2020-06-29 | 2020-08-25 | 扬州大学 | Semen diluent for improving fertilization rate of hatching eggs in later period of egg laying and application thereof |
Non-Patent Citations (1)
| Title |
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| MAROEI MILLAN T.ET AL.: ""Effects of different levels of curcumin antioxidant in beltsville modified diluent on rooster sperm quality after freeze thawing process"", 《ANIMAL SCIENCE RESEARCHES》 * |
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