A combined preparation for improving the conception rate of bovine embryo transplantation and its preparation method and application

Abstract

The present invention relates to the field of animal husbandry and veterinary technology, and specifically to a combined preparation for improving the conception rate of bovine embryo transplantation, and a preparation method and application thereof. The combined preparation comprises preparation A and preparation B; the active ingredients of preparation A are sodium selenite and vitamin E; the active ingredients of preparation B are chorionic gonadotropin and luteinizing hormone releasing hormone A3. By intramuscularly injecting preparation A with active ingredients of sodium selenite and vitamin E into the recipient bovine for embryo transplantation at 8 and 11 months of age, and intramuscularly injecting preparation B with active ingredients of chorionic gonadotropin and luteinizing hormone releasing hormone A3 into the recipient bovine for embryo transplantation on the day of embryo transplantation, the conception rate of bovine embryo transplantation can be improved. When preparation A and preparation B are used together at the same time, the transplantation conception rate can be significantly improved.

Description

Technical Field

[0001] The invention relates to the technical field of animal husbandry and veterinary medicine, and in particular to a combined preparation for improving the conception rate of bovine embryo transplantation, and a preparation method and application thereof. Background Art

[0002] The information disclosed in this background technology section is only intended to enhance the understanding of the overall background of the invention, and should not necessarily be regarded as an admission or any form of suggestion that the information constitutes the prior art already known to a person skilled in the art.

[0003] The cattle industry has experienced two major revolutions. Artificial insemination, especially the popularization of frozen (sex-controlled) semen, is a major revolution in cattle breeding technology, which has played an important role in promoting the development of the cattle industry around the world. Embryo transplantation technology is another revolution in the cattle industry after artificial insemination. Compared with artificial insemination, embryo transplantation technology can fully tap the reproductive potential of excellent female and excellent male animals, shorten the breeding cycle, accelerate genetic progress, reduce the cost of introduction, increase breeding benefits, and reduce the occurrence of infectious diseases.

[0004] Embryo Transfer is the process of removing the early embryo formed after mating of a good-breed female livestock from the uterus or transplanting the embryo obtained through in vitro culture to the corresponding part of the reproductive organ of another (or several) recipient female livestock of the same species that is consistent with the embryo's development stage, so that it can continue to develop, complete the entire growth and development process, and then produce excellent offspring. This is the process commonly known as "pregnancy through borrowing the womb."

[0005] The primary factor limiting the application of bovine in vitro embryo transfer technology is the low embryo transfer conception rate. Therefore, how to further improve the bovine embryo transfer conception rate has become the key to promoting the development of embryo transfer technology. Summary of the invention

[0006] In order to overcome the above problems, the present invention provides a combined preparation for improving the conception rate of bovine embryo transplantation and a preparation method and application thereof.

[0007] The first aspect of the present invention provides a combined preparation for improving the conception rate of bovine embryo transplantation, the combined preparation comprising preparation A and preparation B; the active ingredients of preparation A are sodium selenite and vitamin E; the active ingredients of preparation B are chorionic gonadotropin (HCG) and luteinizing hormone releasing hormone A3 (LHRH-A3).

[0008] Selenium is an essential trace element for animals. It has the functions of anti-oxidation, enhancing immunity, participating in hormone metabolism, and antagonizing toxic metal elements. It plays an important role in the growth and development of animals, especially in the reproductive function of female livestock. Giving appropriate amounts of selenium to female livestock that are deficient in selenium can improve their fertility, the weight and volume of their reproductive organs, and the survival rate and birth weight of young livestock. Vitamin E is an antioxidant that is closely related to the reproductive function of animals. Adding vitamin E can improve the reproductive function of female livestock and reduce the morbidity and mortality of young livestock. Vitamin E and selenium have a certain synergistic effect.

[0009] Chorionic gonadotropin can promote follicle maturation and ovulation, promote corpus luteum formation, stimulate the production and secretion of progesterone, and play an important role in promoting early embryonic development and pregnancy establishment. Luteinizing hormone-releasing hormone A3 can promote the release of luteinizing hormone and follicle-stimulating hormone from the anterior pituitary of animals, which has the effect of promoting ovulation, improving fertilization rate and embryo survival rate.

[0010] In one or more embodiments, both the preparation A and the preparation B are injections.

[0011] Furthermore, the concentration of sodium selenite in the preparation A is 5×10 -4 ~1.5×10 -3 g / mL, and the concentration of vitamin E was 4×10 -2 ~6×10 -2 g / mL;

[0012] Preferably, the concentration of sodium selenite in the preparation A is 1×10 -3 g / mL, and the concentration of vitamin E was 5×10 -2 g / mL.

[0013] Furthermore, the concentration of chorionic gonadotropin in the preparation B is 800-1200 IU / mL, and the concentration of luteinizing hormone-releasing hormone A3 is 10-15 μg / mL;

[0014] Preferably, the concentration of chorionic gonadotropin in the preparation B is 1000 IU / mL, and the concentration of luteinizing hormone releasing hormone A3 is 12.5 μg / mL.

[0015] Furthermore, the auxiliary material of the preparation A and the preparation B is water for injection.

[0016] The second aspect of the present invention provides a method for preparing the above-mentioned combined preparation for improving the conception rate of bovine embryo transplantation, the method comprising:

[0017] Add sodium selenite and vitamin E into water for injection and dissolve them to the required concentration to obtain preparation A;

[0018] Add chorionic gonadotropin and luteinizing hormone releasing hormone A3 into water for injection and dissolve to the required concentration to obtain preparation B.

[0019] The third aspect of the present invention provides the use of the above-mentioned combined preparation for improving the conception rate of bovine embryo transplantation in improving the conception rate of bovine embryo transplantation.

[0020] In one or more embodiments, the embryo transplant recipient cattle are injected intramuscularly with the formulation A at 8 months and 11 months of age, respectively; the embryo transplant recipient cattle are injected intramuscularly with the formulation B on the day of embryo transplant.

[0021] Preferably, the recipient cattle for embryo transplantation are injected intramuscularly with 5 mL of the preparation A at 8 months and 11 months of age, respectively; the recipient cattle for embryo transplantation are injected intramuscularly with 2 mL of the preparation B on the day of embryo transplantation.

[0022] The beneficial effects of the present invention are:

[0023] In the present invention, the embryo transplantation conception rate of cattle can be improved by intramuscularly injecting preparation A, whose effective ingredients are sodium selenite and vitamin E, into the recipient cattle of embryo transplantation at 8 and 11 months of age, and intramuscularly injecting preparation B, whose effective ingredients are chorionic gonadotropin and luteinizing hormone releasing hormone A3, into the recipient cattle of embryo transplantation on the day of embryo transplantation. When preparation A and preparation B are used together, the transplantation conception rate can be significantly improved. DETAILED DESCRIPTION

[0024] It should be noted that the following detailed descriptions are exemplary and are intended to provide further explanation of the present invention. Unless otherwise specified, all technical and scientific terms used in the present invention have the same meanings as those commonly understood by those skilled in the art to which the present invention belongs.

[0025] It should be noted that the terms used herein are only for describing specific embodiments and are not intended to limit exemplary embodiments according to the present invention. As used herein, unless the context clearly indicates otherwise, the singular form is also intended to include the plural form. In addition, it should be understood that when the terms "comprising" and / or "including" are used in this specification, it indicates the presence of features, steps, operations, devices, components and / or combinations thereof.

[0026] The dosage and time mechanism of preparation A and preparation B are adopted in the present invention:

[0027] Cows with a height of ≥135cm, a weight of ≥380kg and above 13 months old can be used for embryo transplantation. Therefore, the embryo transplantation recipient cow must be processed at least before 13 months old. At the same time, because selenium is stored in the liver, multiple continuous injections of sodium selenite can easily cause body poisoning. Therefore, sodium selenite is used in a single injection. After sodium selenite is injected into the body, it reaches its peak in 0.5 hours, and its metabolism rate in the body is slow. Therefore, the injection interval of sodium selenite is 2 to 3 months.

[0028] If preparation B is injected on the day of transplantation, the progesterone content in the blood can reach a high level when the embryo implants, and at the same time maintain a high level of progesterone for a longer period of time, which is beneficial to embryo implantation and subsequent embryo development and prevents miscarriage.

[0029] Preparation of a combined preparation for improving the conception rate of bovine embryo transfer:

[0030] Preparation A: Add the commercially available raw material (>99%) sodium selenite and vitamin E into water for injection. After dissolution, the concentration of sodium selenite is 1×10 -3 g / mL, and the concentration of vitamin E was 5×10 -2 g / mL.

[0031] Preparation B: Add the commercially available raw materials chorionic gonadotropin and luteinizing hormone releasing hormone A3 into water for injection. After dissolution, the concentration of chorionic gonadotropin is 1000 IU / mL, and the concentration of luteinizing hormone releasing hormone A3 is 12.5 μg / mL.

[0032] Conception rate statistics: 30 days after embryo transfer, the recipient cows were tested for pregnancy using B-ultrasound and the embryo transfer conception rate was calculated.

[0033] Example 1 (Determination of the dosage of active ingredient in preparation A)

[0034] When the embryo transplant recipient cows were 11 months old, different effective contents of composition A were injected intramuscularly. The effective contents injected were shown in Table 1. The conception rates were shown in Table 2.

[0035] Table 1 Composition A with different effective contents for intramuscular injection

[0036]

[0037] Table 2 Conception rate

[0038]

[0039] Note: Different superscript letters indicate significant differences (P<0.05)

[0040] As can be seen from Table 2, sodium selenite and vitamin E can improve the conception rate after transplantation when the recipient cows with transplanted bovine embryos are treated at 11 months of age, among which Examples 1 to 5 have the highest conception rate. Therefore, the amount of the active ingredient of the injection composition A can be determined, that is, 5 mL of the injection of the preparation A, in which the concentration of sodium selenite is 1×10 -3 g / mL, and the concentration of vitamin E was 5×10 -2 g / mL.

[0041] Example 2 (Determination of the Optimal Use Time of Preparation A)

[0042] The recipient cows transplanted with bovine embryos were injected intramuscularly with 5 mL of preparation A at 6.5 and 11 months of age, 8 and 11 months of age, and 9.5 and 11 months of age. The concentration of sodium selenite was 1×10 -3 g / mL, and the concentration of vitamin E was 5×10 -2 The injection conditions are shown in Table 3, Table 4 and Table 5, and the conception rates are shown in Table 6, Table 7 and Table 8.

[0043] Table 3 Injection status of recipient cows transplanted with bovine embryos at 6.5 months and 11 months of age

[0044]

[0045]

[0046] Table 4 Injection status of recipient cows transplanted with bovine embryos at 8 and 11 months of age

[0047]

[0048] Table 5 Injection status of recipient cows transplanted with bovine embryos at 9.5 and 11 months of age

[0049]

[0050] Table 6 Conception rate

[0051]

[0052] Note: Different superscript letters indicate significant differences (P<0.05)

[0053] Table 7 Conception rate

[0054]

[0055] Note: Different superscript letters indicate significant differences (P<0.05)

[0056] Table 8 Conception rate

[0057]

[0058] Note: Different superscript letters indicate significant differences (P<0.05)

[0059] As can be seen from Tables 6 to 8, when the recipient cows for transplanted bovine embryos were injected with Preparation A at 8 and 11 months of age, the conception rate after transplantation was significantly improved. Therefore, 8 and 11 months of age are regarded as the best time to use Preparation A.

[0060] Example 3 (Determination of the dosage of active ingredient in preparation B)

[0061] On the day of transplantation, after the embryo transplantation, different effective contents of composition B were injected intramuscularly. The effective contents injected are shown in Table 9, and the conception rates are shown in Table 10.

[0062] Table 9 Composition B with different effective contents for intramuscular injection

[0063]

[0064] Table 10 Conception rate

[0065]

[0066] Note: Different superscript letters indicate significant differences (P<0.05)

[0067] As can be seen from Table 10, the use of HCG and LHRH-A3 to treat recipient cows during embryo transplantation can improve the conception rate after transplantation, among which Example 3-8 has the highest conception rate. Therefore, the amount of the active ingredient of the injection composition B can be determined, that is, 2 mL of preparation B is injected, in which the concentration of chorionic gonadotropin is 1000 IU / mL and the concentration of luteinizing hormone releasing hormone A3 is 12.5 μg / mL.

[0068] Example 4 (Determination of the Optimal Use Time of Preparation B)

[0069] On the day of estrus of the recipient cow and the day of embryo transfer, the recipient cow was injected intramuscularly with 2 mL of preparation B, in which the concentration of chorionic gonadotropin was 1000 IU / mL and the concentration of luteinizing hormone releasing hormone A3 was 12.5 μg / mL. The injection conditions are shown in Table 11, and the conception rate is shown in Table 12.

[0070] Table 11 Injection status of recipient cows on the day of estrus of donor cows and the day of embryo transfer

[0071]

[0072] Table 12 Conception rate

[0073]

[0074] Note: Different superscript letters indicate significant differences (P<0.05)

[0075] It can be seen from Table 12 that the optimal dose of Preparation B injected into the recipient cattle muscle on the day of estrus and the day of embryo transplantation can improve the conception rate after transplantation. At the same time, it was found that Example 4-2 had the highest conception rate, indicating that injecting the optimal dose of Preparation B on the day of embryo transplantation is the best time to use Preparation B.

[0076] Example 5

[0077] The optimal injection doses and injection times of Preparation A and Preparation B determined by Examples 1 to 4 are as follows. In this example, the optimal injection volume and injection time of Preparation A are used, that is, 5 mL of Preparation A is injected, wherein the concentration of sodium selenite is 1×10 -3 g / mL, and the concentration of vitamin E was 5×10 -2 g / mL; 8 months and 11 months of age are the best time to use preparation A. The optimal injection volume and injection time of preparation B is to inject 2mL of preparation B, in which the concentration of chorionic gonadotropin is 1000IU / mL and the concentration of luteinizing hormone-releasing hormone A3 is 12.5μg / mL; the injection on the day of embryo transfer is the best time to use preparation B.

[0078] The recipient cows were treated as shown in Table 13, wherein a Preparation A group was set up using the optimal injection volume and injection time of Preparation A, and a Preparation B group was set up using the optimal injection volume and injection time of Preparation B. The conception rates are shown in Table 14.

[0079] Table 13 Treatment of recipient cattle

[0080]

[0081] Table 14 Conception rate

[0082]

[0083] Note: Different superscript letters indicate significant differences (P<0.05)

[0084] As shown in Table 13, Preparation Group A and Preparation Group B can improve the conception rate. When Preparation Group A and Preparation Group B are treated together, the effect is best and can significantly improve the transplantation conception rate.

[0085] The above description is only a preferred embodiment of the present invention and is not intended to limit the present invention. For those skilled in the art, the present invention may have various modifications and variations. Any modification, equivalent replacement, improvement, etc. made within the spirit and principle of the present invention shall be included in the protection scope of the present invention.

Claims

1. The application of a combined preparation for improving the conception rate of bovine embryo transplantation in improving the conception rate of bovine embryo transplantation is characterized in that: The recipient cattle for embryo transplantation are injected intramuscularly with the preparation A at 8 months and 11 months of age respectively; the recipient cattle for embryo transplantation are injected intramuscularly with the preparation B on the day of embryo transplantation; The combined preparation for improving the conception rate of bovine embryo transplantation comprises preparation A and preparation B; the active ingredients of preparation A are sodium selenite and vitamin E; the active ingredients of preparation B are chorionic gonadotropin and luteinizing hormone releasing hormone A3.

2. The use according to claim 1, characterized in that The preparations A and B are both injections.

3. The use according to claim 2, characterized in that The concentration of sodium selenite in the preparation A is 5×10 -4 ~1.5×10 -3 g / mL, and the concentration of vitamin E was 4×10 -2 ~6×10 -2 g / mL; The concentration of chorionic gonadotropin in the preparation B is 800-1200 IU / mL, and the concentration of luteinizing hormone releasing hormone A3 is 10-15 μg / mL.

4. The use according to claim 3, characterized in that The concentration of sodium selenite in the preparation A is 1×10 -3 g / mL, and the concentration of vitamin E was 5×10 -2 g / mL; The concentration of chorionic gonadotropin in the preparation B is 1000 IU / mL, and the concentration of luteinizing hormone releasing hormone A3 is 12.5 μg / mL.

5. The use according to claim 2, characterized in that The auxiliary materials of the preparation A and the preparation B are water for injection.

6. The use according to claim 1, characterized in that Add sodium selenite and vitamin E into water for injection and dissolve them to the required concentration to obtain preparation A; Add chorionic gonadotropin and luteinizing hormone releasing hormone A3 into water for injection and dissolve to the required concentration to obtain preparation B.

7. The use according to claim 1, characterized in that The recipient cattle for embryo transplantation were injected intramuscularly with 5 mL of the preparation A at 8 and 11 months of age, respectively; the recipient cattle for embryo transplantation were injected intramuscularly with 2 mL of the preparation B on the day of embryo transplantation.

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