CN111406738A - Chicken essence diluent for artificial oviduct insemination and preparation method thereof - Google Patents
Chicken essence diluent for artificial oviduct insemination and preparation method thereof Download PDFInfo
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- CN111406738A CN111406738A CN202010387371.5A CN202010387371A CN111406738A CN 111406738 A CN111406738 A CN 111406738A CN 202010387371 A CN202010387371 A CN 202010387371A CN 111406738 A CN111406738 A CN 111406738A
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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 technical field of poultry breeding, and discloses a chicken sperm diluent for artificial oviduct insemination and a preparation method thereof, wherein the chicken sperm diluent comprises potassium citrate, fructose, sodium glutamate, potassium dihydrogen phosphate, dipotassium hydrogen phosphate, magnesium chloride hexahydrate, TES, sodium acetate and deionized water, the survival rate of chicken sperm preserved for 2 hours in the sperm diluent is still higher and is more than 50 percent, cock sperm can be preserved at normal temperature by using the sperm diluent, the time of artificial insemination of chickens can be prolonged, and the utilization rate of the cock sperm can be favorably prolonged7～2.1×107In between, insemination and hatchling may be achieved.
Description
Technical Field
The invention belongs to the technical field of poultry breeding, and particularly relates to chicken essence diluent for artificial oviduct insemination and a preparation method thereof.
Background
Artificial insemination techniques are the artificial introduction of sperm into the cervix or uterine cavity of a female animal in order to achieve in vivo fertilization by means other than natural mating, in order to overcome the lack of natural mating capacity or to accomplish the fertility aspects of donated sperm (Magyar, 1991). With the improvement of the quality and living standard of modern people, the supply demand of poultry products is increased year by year, and a plurality of developed countries of the poultry industry explore a new artificial insemination technology in a large scale so as to improve the production efficiency and reduce the insemination cost. With the continuous development of human society and the continuous development and utilization of natural environment, the habitat of birds is continuously contracted, resulting in more and more birds being listed on lists of endangered species (Moorman et al, 2001). Cage culture protection becomes a main breeding means of rare birds, but the hatching egg shell breaking rate of cage culture protection of wild birds is low. Artificial insemination methods for poultry are widely used for the reproduction of non-poultry birds, particularly endangered birds (Sen, 2004).
The semen diluent is essential to the preservation, transportation and insemination of chicken sperms, can obviously reduce the breeding cost of cocks and the utilization rate of sperms, and improves economic benefits. At present, the domestic chicken essence diluent is only Beijing poultry diluent (BJJX). For the differences of the quality and the feeding environment of the researched chickens, the insemination parts of the semen liquid are different, and the developed semen diluent cannot necessarily achieve the expected effect.
Disclosure of Invention
In order to solve the defects and shortcomings of the prior art, the invention provides a chicken essence diluent for artificial oviduct insemination.
The invention also aims to provide a preparation method of the chicken essence diluent for artificial oviduct insemination.
The purpose of the invention is realized by the following technical scheme:
a chicken essence diluent for artificial oviduct insemination comprises potassium citrate, fructose, sodium glutamate, potassium dihydrogen phosphate, dipotassium hydrogen phosphate, magnesium chloride hexahydrate, TES, sodium acetate and sterilized deionized water.
Preferably, each 1L sterilized deionized water contains 5.5-6.5 g of potassium citrate, 0.04-0.06 g of fructose, 7.8-9.6 g of sodium glutamate, 0.6-0.8 g of monopotassium phosphate, 8.7-11 g of dipotassium phosphate, 0.3-0.4 g of magnesium chloride hexahydrate, 2.7-3.3 g of TES and 2.3-2.9 g of sodium acetate.
More preferably, the sterilized deionized water of 1L contains 6g of potassium citrate, 0.05g of fructose, 8.7g of sodium glutamate, 0.65g of monopotassium phosphate, 9.7g of dipotassium phosphate, 0.34g of magnesium chloride hexahydrate, 3g of TES and 2.6g of sodium acetate.
The preparation method of the chicken essence diluent for artificial oviduct insemination comprises the steps of mixing potassium citrate, fructose, sodium glutamate, potassium dihydrogen phosphate, dipotassium hydrogen phosphate, magnesium chloride hexahydrate, TES and sodium acetate, adding deionized water, fully stirring until the mixture is completely dissolved, and fixing the volume to 1L to prepare the semen diluent.
The application of the chicken essence diluent for artificial oviduct insemination is provided.
Preferably, when the artificial oviduct is inseminated, the number of sperms injected each time is (1.8-2.1) × 107And (4) respectively.
Compared with the prior art, the invention has the following beneficial effects:
1. the survival rate of the chicken sperm preserved for 2 hours in the sperm diluent is still higher and is more than 50 percent. The semen diluent can be used for preserving the cock sperms at normal temperature, and is beneficial to prolonging the artificial insemination time of the chickens and the utilization rate of the cock sperms.
2. The sperm diluent is used for laparoscope-mediated artificial oviduct insemination, and the injected sperm quantity is ensured to be 1.8 × 10 in each artificial oviduct insemination7～2.1×107And insemination and hatching can be realized.
Detailed Description
The following examples are presented to further illustrate the present invention and should not be construed as limiting the invention. Unless otherwise specified, the technical means used in the examples are conventional means well known to those skilled in the art.
Example 1
1. Under aseptic conditions, potassium citrate, fructose, sodium glutamate, monopotassium phosphate, dipotassium phosphate, magnesium chloride hexahydrate, TES and sodium acetate are respectively weighed according to the formula in Table 1, 100ml of sterilized distilled water is added into a beaker, the mixture is fully stirred until the mixture is completely dissolved, the mixture is transferred into a 1L volumetric flask, the volume is constant to 1L, the solution is filtered by a 0.45 mu m disposable filter, and a semen diluent is prepared and stored at 4 ℃.
2. The chicken semen diluent comprises potassium citrate (K)3C6H5O7) Fructose (C)6H12O6) Sodium glutamate (C)5H8NNaO4) Potassium dihydrogen phosphate (KH)2PO4) Dipotassium hydrogen phosphate (K)2HPO4) Magnesium chloride hexahydrate (MgCl)2·6H2O)、TES(C6H15O3Si) and sodium acetate (CH)3COONa)。
TABLE 1 ingredient list of diluted semen of cock
3. The storage method comprises the following steps: table 2 shows the effect of semen dilution on chicken semen survival (%) at 37 ℃. The semen diluent is used for preserving the chicken sperms at 37 ℃, and the survival rate of the sperms of the chicken sperms preserved for 2 hours in the semen diluent is still higher and is more than 50 percent as can be seen from the table 2. Therefore, the semen diluent can be used for preserving the cock sperms at normal temperature, and is beneficial to prolonging the artificial insemination time of the chickens and the utilization rate of the cock sperms. When in use, the semen diluent is preheated to 37 ℃ in advance, then is mixed with cock semen for use, and the dosage of the semen diluent can be increased or reduced according to the required semen concentration.
Effect of semen dilution on the survival (%) of chickens at 237 ℃ in Table
4. The application method comprises observing sperm motility count by semen microscopy, adding semen diluent, and diluting to about 2 × 107M L, the number of injected sperms is 1.8 × 10 during each artificial insemination of the oviduct7～ 2.1×107In the meantime. Subjecting hen to be fertilized to wingAfter the lower vein anesthesia, the chicken sperm diluted by the semen diluent is injected into the oviduct of the hen to be fertilized by using a laparoscope for mediation, and then the laying rate and the hatching rate of the hen are observed as shown in table 3. Table 3 shows the fertilization rate of artificial oviduct insemination. As can be seen, the sperm diluent is used for laparoscopy-mediated artificial oviduct insemination, and insemination and hatching can be realized.
The molar concentration ratio of sodium to potassium in the semen diluent of the invention is 1: 2, the content of potassium is far higher than that of chicken semen deposition liquid Na+And K+Molar concentration ratio of 15: 1, i.e. Na is contained in 10ml of seminiferous duct fluid+39.3mg， K+4.3mg。K+Is essential to maintaining sperm volume, semen pH and obtaining and maintaining sperm motility. A small amount of K+Can maintain the normal function and necessary activity of the sperms; in addition, an appropriate amount of K+The potassium citrate in the semen diluent provides a weakly acidic environment for the semen, continuously provides bioavailable potassium ions in a cationic form, can prolong the life of the semen and improve the survival index of the semen, and keeps the sperm aberration rate at a lower level compared with inorganic salts such as sodium chloride, potassium chloride and the like, so that the sperm diluent is used for laparoscopically mediating the artificial oviduct, and the quantity of injected sperms is ensured to be 1.8 × 10 during each artificial oviduct insemination7～2.1×107In between, insemination and hatchling may be achieved.
TABLE 3 fertilization rates for artificial oviduct insemination
Example 2
1. Under the aseptic condition, 5.5g of potassium citrate, 0.04g of fructose, 8.5g of sodium glutamate, 0.6g of monopotassium phosphate, 9.5g of dipotassium phosphate, 0.3g of magnesium chloride hexahydrate, 2.5g of TES and 2.5g of sodium acetate are respectively weighed, 100ml of sterilized distilled water is added into a beaker, the mixture is fully stirred until the mixture is completely dissolved, the beaker is transferred into a volumetric flask of 1L, the volume is fixed to 1L, and then the solution is filtered by a 0.45 mu m disposable filter to prepare a semen diluent which is stored at the temperature of 4 ℃.
2. When in use, the semen diluent is preheated to 37 ℃ in advance, then is mixed with cock semen for use, and the dosage of the semen diluent can be increased or reduced according to the required semen concentration.
Example 3
1. Under aseptic conditions, 6.5g of potassium citrate, 0.06g of fructose, 9.0g of sodium glutamate, 0.7g of monopotassium phosphate, 10g of dipotassium phosphate, 0.4g of magnesium chloride hexahydrate, 3.5g of TES and 2.8g of sodium acetate are weighed according to the formula in Table 1, 100ml of sterilized distilled water is added into a beaker, the mixture is fully stirred until the mixture is completely dissolved, the mixture is transferred into a volumetric flask of 1L, the volume is fixed to 1L, the solution is filtered by a 0.45 mu m disposable filter to prepare a semen diluent, and the semen diluent is stored at 4 ℃.
2. When in use, the semen diluent is preheated to 37 ℃ in advance, then is mixed with cock semen for use, and the dosage of the semen diluent can be increased or reduced according to the required semen concentration.
The above embodiments are preferred embodiments of the present invention, but the present invention is not limited to the above embodiments, and any other changes, modifications, substitutions, combinations and simplifications which do not depart from the spirit and principle of the present invention should be construed as equivalents thereof, and all such changes, modifications, substitutions, combinations and simplifications are intended to be included in the scope of the present invention.
Claims (6)
1. The chicken essence diluent for artificial oviduct insemination is characterized by comprising potassium citrate, fructose, sodium glutamate, monopotassium phosphate, dipotassium phosphate, magnesium chloride hexahydrate, TES, sodium acetate and sterilized deionized water.
2. The chicken essence diluent for artificial insemination of an oviduct of claim 1, wherein each 1L sterilized deionized water contains 5.5-6.5 g of potassium citrate, 0.04-0.06 g of fructose, 7.8-9.6 g of sodium glutamate, 0.6-0.8 g of potassium dihydrogen phosphate, 8.7-11 g of dipotassium hydrogen phosphate, 0.3-0.4 g of magnesium chloride hexahydrate, 2.7-3.3 g of TES, and 2.3-2.9 g of sodium acetate.
3. The chicken essence diluent for artificial insemination of an oviduct according to claim 2, wherein each 1L sterilized deionized water contains 6g of potassium citrate, 0.05g of fructose, 8.7g of sodium glutamate, 0.65g of potassium dihydrogen phosphate, 9.7g of dipotassium hydrogen phosphate, 0.34g of magnesium chloride hexahydrate, 3g of TES and 2.6g of sodium acetate.
4. The method for preparing chicken essence diluent for artificial oviduct insemination according to any one of claims 1 to 3, wherein the method comprises the steps of mixing potassium citrate, fructose, sodium glutamate, potassium dihydrogen phosphate, dipotassium hydrogen phosphate, magnesium chloride hexahydrate, TES and sodium acetate, adding deionized water, fully stirring until the mixture is completely dissolved, and then fixing the volume to 1L to obtain the semen diluent.
5. Use of a dilution of chicken essence according to any one of claims 1 to 3 for artificial oviduct insemination.
6. The use of the chicken essence diluent for artificial oviduct insemination according to claim 5, wherein the number of sperms injected each time is (1.8-2.1) × 107And (4) sperms.
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| CN202010387371.5A CN111406738A (en) | 2020-05-09 | 2020-05-09 | Chicken essence diluent for artificial oviduct insemination and preparation method thereof |
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Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN114375946A (en) * | 2022-01-22 | 2022-04-22 | 上海市农业科学院 | Cryopreservation method of chicken semen |
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| CN104094926A (en) * | 2014-08-05 | 2014-10-15 | 燕海峰 | Chicken semen diluent and preparation and utilization methods thereof |
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2020
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| CN114375946A (en) * | 2022-01-22 | 2022-04-22 | 上海市农业科学院 | Cryopreservation method of chicken semen |
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