RU2823019C1 - Method for superovulation in embryo donor cows - Google Patents

Abstract

FIELD: biotechnology.

SUBSTANCE: invention relates to and represents a method for superovulation in embryo donor cows, including introduction on 10th, 11th and 13th day of the sexual cycle twice intramuscularly of the preparation "ФСГ-супер" in dosage of 6AE, on 12th day of the sexual cycle, prostaglandin F2α is added to the preparation "ФСГ-супер" in dosage of 500 mcg at 8–00 and in dosage of 250 mcg at 20–00, on 14th day of the sexual cycle, at the end of oestrus, the preparation is injected intramuscularly, wherein the preparation is represented by Fermivit-Se in a single dose of 8 ml/head.

EFFECT: invention makes it possible to increase efficiency of superovulation in cattle.

1 cl, 1 tbl, 1 ex

Description

The invention relates to biotechnology and makes it possible to increase the efficiency of superovulation in cattle.

Methods for performing superovulation in cows using gonadotropins are widely known.

A commonly used gonadotropin for superovulation is FFA. To ensure the growth of several follicles and their subsequent ovulation, FFA is injected 4-5 days before the expected heat. This is achieved by starting hormonal treatment of the donor 2-3 days before the natural or induced regression of the corpus luteum. In the first case, the drug is administered during the transition period from the luteal to the follicular phase of the reproductive cycle on days 16-17 and then, when the first signs appear and during the expected heat, 2000 IU hCG is injected intravenously (3).

The advantage of this method is the low cost of FFA and the ability to obtain the drug directly at the enterprise that produces embryos. The disadvantage of this method is the high anaphylactogenicity of FFA. In addition, the European Parliament intends to introduce a ban on the import of pregnant mares blood serum (MAB), which increases fertility in animals (5).

The advantage of this method is the single use of gonadotropin. However, this method requires the additional use of estradiol benzoate and human chorionic gonadotropin. This method is extended over a time frame, which causes an increase in the cost of obtaining embryos due to feeding and maintenance of donors (1).

The advantage of this method is the short period of hormonal treatment of the embryo donor animal. However, the prolonged action of gonadotropins does not allow obtaining a lasting result in the form of superovulation and embryo quality (1).

The advantage of this method is the uniform effect of the drug on the ovary and the production of a large number of embryos suitable for freezing and storage. The disadvantage is the high cost of the drug (4).

There is a known method for inducing superovulation in embryo donor cows with prolongation of the action of pituitary gonadotropins, which includes a single injection of a drug containing pituitary gonadotropins follicle stimulating hormone (FSH) and luteinizing hormone (LH) into donor cows in the luteal phase of the reproductive cycle, followed by administration after 48 hours prostaglandin F2α. In the method, a 0.25-1% solution of novocaine is additionally administered in an amount of 3-15 ml, and this solution of novocaine is administered simultaneously in combination with a drug containing the pituitary gonadotropins FSH and LH, combined with a prolongator, while a biologically degradable polymer is used as a prolongator polyethylene glycol (PEG) with a mass of 6000 to 70000 Da in an amount of 0.5-7 g, and a drug containing pituitary gonadotropins FSH and LH is administered in the total dose recommended by the instructions for this drug for inducing superovulation (RF patent No. 2617042, 2017 .)

The disadvantage of this method, according to many experts, is: “Side effects of novocaine, and the most typical side effects are:

• numbness;

• tingling;

• slight pain at the injection site.

Novocaine is low-toxic, but often causes allergic reactions. Their manifestations may be:

• stomatitis;

• dermatitis;

• Quincke's edema;

• anaphylactic shock.

With increased sensitivity to novocaine, the patient experiences:

• dizziness;

• weakness;

• drop in blood pressure;

• collapse;

• shock.

Other unusual side effects include chest pain, decreased heart rate, drowsiness, restlessness, nausea, vomiting, shaking, seizures" (https://www.colgate.ru/oral-health/anesthesia/what-are-common-novocain -side-effects).

The closest technical solution is the method of performing superovulation using the drug “FSH-super” (see RF patent for invention No. 2469738, class A61D 19/02, 2012), in which to stimulate polyovulation in embryo donor cows, starting from the 9-10th day of the sexual cycle, they are administered the drug “FSH-super” at a dose of 6 units. Armor standard (AE) (120 I.E.) twice a day every 12 hours. On the third day after the start of administration, one of the F2α drugs (estrophan, clatraprostin, etc.) is administered simultaneously with FSH-super, one dose in the morning and in the evening. Detection of signs of heat in cows begins 36-48 hours after the administration of prostaglandin F2α. As the animals come into heat, they are artificially inseminated 2-3 times with an interval of 12 hours. Embryos from donor cows are removed within the time period necessary to obtain embryos of the desired stages (on days 7-8), considering the day of the first insemination of animals as day zero, using instructions for the use of “FSH-super” (2).

The advantage of this method is the low cost of the drugs used and the high yield of embryos.

The disadvantage of this method is its lack of effectiveness due to the lack of nonspecific stimulation of the ovaries and uterus.

The technical result of the invention is to increase the efficiency of superovulation in embryo donor cows and improve their quality due to a nonspecific effect on the ovaries and uterus of the animal.

The technical result is achieved by the fact that in the method of superovulation in embryo donor cows, including the administration of the FSH-super drug twice intramuscularly on the 10th, 11th and 13th days of the sexual cycle at a dosage of 6AE, on the 12th day of the sexual cycle prostaglandin F2α is added to the drug “FSH-super” at a dosage of 6AE at a dosage of 500 mcg at 8-00 and at a dosage of 250 mcg at 20-00, on the 14th day of the sexual cycle, at the end of sexual heat, the drug is administered intramuscularly, characterized in that Fermivit-Se is used as a drug, at a dose of 8 ml/head once.

A comparative analysis of the proposed and used methods of superovulation in embryo donor cows allows us to identify features that distinguish the proposed technical solution from the prototype: increasing the effectiveness of the influence of follicle-stimulating hormone during superovulation, as well as improving the quality of the resulting embryos - is achieved by intramuscular injection of the vitamin and mineral preparation "Fermivit-Se" " The injection of the drug is carried out on the day of artificial insemination in a dose of 7-8 ml per head once.

To date, from the patent and scientific and technical literature available to the authors, the combined use of Fermivit-Se with follicle-stimulating hormone, which ensures an increase in the efficiency of superovulation in embryo donor cows in order to obtain a larger number of viable embryos, is unknown.

Thus, the proposed method of performing superovulation in embryo donor cows meets the “novelty” criterion.

The claimed method has an inventive step, since for the average specialist it does not clearly follow from the state of the art: neither the instructions nor the recommendations for embryo transplantation include the use of vitamin and mineral preparations, in particular Fermivit-Se.

The method is industrially applicable, as it can be used to stimulate superovulation in donor cows.

Fermivit-Se - solution for injection, as active ingredients in 1 ml contains ferric iron (in complex with low molecular weight dextran) - 40 mg, vitamin A - 2500 IU, vitamin D3-500 IU, vitamin E - 2.5 mg, vitamin K - 0.25 mg, vitamin B1 - 2.5 mg, vitamin B2 - 1 mg, vitamin B6 - 0.75 mg, vitamin B12 - 0.0025 mg, nicotinamide - 7.5 mg, pantothenic acid - 5 mg, folic acid - 0.05 mg, biotin - 0.0025 mg, sodium selenite (in terms of selenium) -0.15 mg, lactoalbumin protein hydrolyzate - 0.2 mg, glucose - 15 mg, and nipagin as excipients - 0.5 mg, water for injection - up to 1 ml. Fermivit-Se is prescribed for the treatment and prevention of hypovitaminosis and animal diseases caused by a lack of iron and selenium, as well as for stress, disorders of reproductive function, retardation of young animals in growth and development, during the period of vaccination, deworming, recovery after illnesses (https:// www.vidal.ru/veterinar/fermivit-se-30181).

The method of superovulation in embryo donor cows is carried out as follows: on the 10th, 11th and 13th days of the sexual cycle, the drug “FSH-super” is administered intramuscularly twice at a dosage of 6 AE (at ^8:00 and ^20:00 ). On the 12th day of the sexual cycle, prostaglandin F2α is added to the FSH-super drug at a dosage of 6 AE at a dosage of 500 mcg at ^{8:00 am} and at a dosage of 250 mcg at ^{20:00 am} . On the 14th day of the reproductive cycle, at the end of heat, the drug “Fermivit-Se” is administered intramuscularly at a dose of 8 ml/animal and the animal is inseminated. On the 21st day of the sexual cycle, non-surgical embryo removal is performed. The administered dose of 8 ml/bird of the Fermivit-Se drug is due to the fact that if you take less than 8 ml/bird, the effectiveness will be low, and if the dose of the drug is more than 8 ml/bird, side effects may occur.

An example of a specific implementation of the proposed method

To implement the proposed method of performing superovulation in embryo donor cows, scientific and production experience was carried out.

In the conditions of the educational and experimental farm “Kuban” of the Federal State Budgetary Educational Institution of Higher Education Kuban, Krasnodar Territory, according to the principle of pair-analogues, two groups of cows of 15 heads each were formed: a control (prototype) and an experimental (the claimed method).

The table shows that 0.2 more embryos were obtained from cows from the experimental group than in the control group (according to the prototype). Moreover, the number of embryos suitable for transplantation in the experimental group is 0.3 embryos more than in the control group (according to the prototype). Data are given per animal.

Thus, the proposed method makes it possible to obtain a larger number of viable embryos from donor cows.

Literature

1. Valyushkin K.D., Medvedev G.F. Obstetrics, gynecology and biotechnology of animal reproduction: Textbook, 2nd ed., revised. And additional - Mn.: Urajai, 2001. - P. 320.

2. Instructions for the use of “FSH-super” to stimulate ovarian function in female animals and polyovulation in embryo donor cows. Approved by Deputy Head of the Veterinary Department B.A. Kobzev April 23, 1996 Registration number of the drug PVR-2-1.0/00269.

3. Prokofiev M.I. Regulation of reproduction of farm animals. - L.: Science, 1983. - P. 182.

4. Ernst L.K., Sergeev N.I. Embryo transplantation of farm animals. - M.: Agropromizdat, 1989. - P. 117.

5. https://www.partyfortheanimals.com/ru/eu-parliament-opposes-livestock-medicine-of-horse-blood.

Claims (1)

A method of performing superovulation in embryo donor cows, including the administration of the FSH-super drug twice intramuscularly at a dosage of 6AE on the 10th, 11th and 13th days of the reproductive cycle, at a dosage of 6AE, on the 12th day of the sexual cycle to the FSH-super drug at a dosage of 6AE, prostaglandin F2α is added at a dosage of 500 mcg at 8-00 and at a dosage of 250 mcg at 20-00, on the 14th day of the sexual cycle, at the end of heat, a drug is administered intramuscularly, characterized in that the drug used is “Fermivit- Se", at a dose of 8 ml/animal once.