CN111568600B - Method for improving availability of acceptor cattle in cattle embryo transplantation - Google Patents
Method for improving availability of acceptor cattle in cattle embryo transplantation Download PDFInfo
- Publication number
- CN111568600B CN111568600B CN202010467616.5A CN202010467616A CN111568600B CN 111568600 B CN111568600 B CN 111568600B CN 202010467616 A CN202010467616 A CN 202010467616A CN 111568600 B CN111568600 B CN 111568600B
- Authority
- CN
- China
- Prior art keywords
- oestrus
- cattle
- cows
- injection
- cow
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Active
Links
Classifications
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61D—VETERINARY INSTRUMENTS, IMPLEMENTS, TOOLS, OR METHODS
- A61D19/00—Instruments or methods for reproduction or fertilisation
- A61D19/04—Instruments or methods for reproduction or fertilisation for embryo transplantation
-
- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01K—ANIMAL HUSBANDRY; CARE OF BIRDS, FISHES, INSECTS; FISHING; REARING OR BREEDING ANIMALS, NOT OTHERWISE PROVIDED FOR; NEW BREEDS OF ANIMALS
- A01K67/00—Rearing or breeding animals, not otherwise provided for; New breeds of animals
- A01K67/02—Breeding vertebrates
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K38/00—Medicinal preparations containing peptides
- A61K38/16—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
- A61K38/17—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans
- A61K38/22—Hormones
- A61K38/24—Follicle-stimulating hormone [FSH]; Chorionic gonadotropins, e.g. HCG; Luteinising hormone [LH]; Thyroid-stimulating hormone [TSH]
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K47/00—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient
- A61K47/06—Organic compounds, e.g. natural or synthetic hydrocarbons, polyolefins, mineral oil, petrolatum or ozokerite
- A61K47/08—Organic compounds, e.g. natural or synthetic hydrocarbons, polyolefins, mineral oil, petrolatum or ozokerite containing oxygen, e.g. ethers, acetals, ketones, quinones, aldehydes, peroxides
- A61K47/14—Esters of carboxylic acids, e.g. fatty acid monoglycerides, medium-chain triglycerides, parabens or PEG fatty acid esters
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K47/00—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient
- A61K47/06—Organic compounds, e.g. natural or synthetic hydrocarbons, polyolefins, mineral oil, petrolatum or ozokerite
- A61K47/16—Organic compounds, e.g. natural or synthetic hydrocarbons, polyolefins, mineral oil, petrolatum or ozokerite containing nitrogen, e.g. nitro-, nitroso-, azo-compounds, nitriles, cyanates
- A61K47/18—Amines; Amides; Ureas; Quaternary ammonium compounds; Amino acids; Oligopeptides having up to five amino acids
- A61K47/183—Amino acids, e.g. glycine, EDTA or aspartame
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K9/00—Medicinal preparations characterised by special physical form
- A61K9/0012—Galenical forms characterised by the site of application
- A61K9/0019—Injectable compositions; Intramuscular, intravenous, arterial, subcutaneous administration; Compositions to be administered through the skin in an invasive manner
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K9/00—Medicinal preparations characterised by special physical form
- A61K9/14—Particulate form, e.g. powders, Processes for size reducing of pure drugs or the resulting products, Pure drug nanoparticles
- A61K9/19—Particulate form, e.g. powders, Processes for size reducing of pure drugs or the resulting products, Pure drug nanoparticles lyophilised, i.e. freeze-dried, solutions or dispersions
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P15/00—Drugs for genital or sexual disorders; Contraceptives
- A61P15/08—Drugs for genital or sexual disorders; Contraceptives for gonadal disorders or for enhancing fertility, e.g. inducers of ovulation or of spermatogenesis
Abstract
The invention discloses a method for improving the availability of a receptor cow in cow embryo transplantation. The present invention achieves the technical result of one or more aspects of the present invention by injecting luteinizing hormone releasing hormone a3 into a cow. The method comprises the following steps: (1) providing a donor bovine; (2) intramuscular injection of luteinizing hormone releasing hormone a3 in donor cattle; (3) continuously observing the oestrus and ovulation conditions of the cows within 3d from the time of injecting the medicaments, recording the oestrus starting time and the oestrus ending time of the cows, observing the oestrus starting time and the oestrus ending time once every morning and evening, determining the oestrus time, the oestrus program and the ovulation condition by combining rectal examination according to the appearance oestrus symptoms of the cows, determining the oestrus appearing within 3d after injecting the medicaments as oestrus synchronization, and carrying out artificial insemination on the cows in oestrus and ovulation at the right time according to a freeze-matching technical operation rule, wherein semen is the frozen semen of the cows of the same donor; (4) and (5) determining whether the cow is pregnant according to the rectal examination result 90 days after mating, and feeding the cow in the pregnancy period according to a conventional method.
Description
Technical Field
The invention belongs to the field of livestock raising, relates to a cattle breeding technology, and particularly relates to a method for improving the availability of a receptor cattle in cattle embryo transplantation. The present invention achieves the technical effect of one or more aspects of the present invention by injecting luteinizing hormone releasing hormone A3, LHRH-A3, into cows.
Background
At present, when a recipient cow is selected in production, the recipient cow has two schemes of natural estrus or synchronous estrus by injecting medicaments, and the latter scheme plays a very important role in improving the production efficiency.
Estrus synchronization refers to the use of certain hormone preparations to artificially control and regulate the progress of the estrus cycle of a group of dam animals so that they will have a concentrated estrus within a predetermined time. Oestrogen inhibiting agents such as pregnenones. The control method includes prostaglandin method and progestogen method. Generally, estrus synchronization refers to a technique of regulating the estrus cycle of a dam using a hormone preparation to allow it to estrus in a relatively concentrated time. Commonly used estrus-synchronization drugs, one type is agents that inhibit estrus, such as pregnenones, during their treatment, increase progesterone levels in the dams at follicular stage, inhibit the maturation of the follicles, and stop natural estrus, actually prolonging the estrus cycle. After a certain period of time, the drug treatment was stopped at the same time, so that progesterone levels were reduced and follicles in the treated dams began to grow rapidly and appeared at the same time.
Estrus synchronization is a new technology for controlling estrus. The research of all countries in the world is started from the 60 s, and the research also shows the power of the research on breeding and improving cattle in all countries along with the deep research. Therefore, China also starts research in seventy-eight years, and a plurality of units carry out tests and obtain certain results. A group of cows is treated by medicinal hormones, the natural oestrus cycle of the group of cows is consciously adjusted and changed, the dispersed oestrus of each cow is adjusted to a certain time, the whole group of cows centrally and uniformly oestrus, centralized breeding is realized, and the production efficiency can be greatly improved.
The importance of estrus synchronization is manifold. For example, 1, estrus, mating, pregnancy and delivery of a herd can be adjusted to be performed simultaneously within a certain time, so that the advantages that calves are born simultaneously, management and cultivation in the lactation period can be unified, scientific feeding management of the calves and the grown-up calves can be intensively done on a large scale, and the defects that a plurality of devices are utilized and the work of personnel is uneven throughout the year are overcome; 2. the frozen semen and the artificial insemination are technical works, and the efficiency and the working quality can be improved by concentrating the strength. The timing insemination can be carried out without catching estrus for identification; 3. meanwhile, oestrus, superovulation and embryo transplantation are several important links of a new propagation technology, but only by the way, the embryo transplantation can be realized only when a donor cow and a receptor cow oestrate simultaneously and genitals are in the same physiological state.
The simultaneous estrus is based on the physiological mechanism of animals as a theoretical basis. The shape and function of the ovary play an important role in the reproductive physiology of cows, and the ovary passes through two stages, namely a follicular stage and a luteal stage in the oestrus cycle of cows. The two phases are alternated and appear repeatedly to form an estrus cycle. Control of the luteal phase of both phases is critical to the control of the estrus cycle. Each cow in the cow group is in different stages of the estrus cycle, and the control of estrus is to control the life of corpus luteum in the corpus luteum through hormone or drug treatment, so that the estrus cycles of all cows are adjusted to the same stage, and the purpose of synchronous estrus is achieved. There are two main approaches to luteal control, one is the administration of progestins and the other is the administration of prostaglandins to interrupt the natural estrous cycle of the cow. The effect of using the progestogen progesterone and analogues thereof is that the continuous use of the progestogen in the blood maintains a certain level, inhibits the growth and development and the estrus of follicles on ovaries, and is always in the artificial corpus luteum stage, sometimes the corpus luteum on the ovaries has disappeared, which means that the endogenous hormone level is reduced, but as the exogenous hormone is still acting, the cows will not estre, which is equivalent to prolonging the estrus cycle and postponing the estrus. The second approach is to use prostaglandins such as prostaglandin F2. And the like. Aims to dissolve corpus luteum, reduce the level of progesterone, stop the luteal phase, promote the release of pituitary gonadotropin and ensure the estrus to come. Therefore, the estrus cycle is shortened and the estrus comes in advance.
There are several references to methods of improving reproductive performance in animals using luteinizing hormone releasing hormone A3. For example, CN101711513A (application No. 200910272961.7) discloses a method for increasing the ovulation number of sows and an injection thereof, belonging to the technical field of agricultural biological products. The method is characterized in that PG (sodium chlorophrostate) and PMSG (serum gonadotropin) are injected simultaneously before estrus, or a mixture of PG and PMSG is injected, HCG (chorionic gonadotropin) or LHRH-A3 (luteinizing hormone releasing hormone A3) is injected during estrus, the effect of promoting the ovulation number of young sows or multiparous sows to reach 2-3 times of the normal egg discharge amount is achieved, the reproductive performance of the sows is improved, the number of sows is obviously increased, and the method is stable in technology and obvious in effect; does not induce ovarian cyst, and can achieve the same superovulation effect on replacement gilts by using the method at any time before sexual maturity.
CN103083110A (application No. 201210578058.5) discloses a method for improving the estrus rate and pregnancy rate of embryo transplantation recipient cattle at the same period, vitamin ADE10 ml is injected into the recipient cattle with or without cycle, a progestogen vaginal suppository is placed for 9-12 days, 1000 units of progestogen are injected into the pregnant horse in intramuscular mode 1 day before suppository injection, and 0.5 mg of chloroprostenol is injected into the pregnant horse in suppository injection. Taking out and oestrus 2-3 days after the embolism, injecting 2000 units of chorionic hormone or 25 micrograms of No. 3 chorionic hormone into the cattle, injecting an anti-inflammatory drug into the uterus of a receptor cattle which has oestrous and has slight metritis after treatment, purifying the uterus on the same day, and injecting 10 milliliters of vitamin ADE into the uterus, so that the intrauterine environment of the receptor cattle is more favorable for pregnancy. Embryo transfer is carried out on the 7 th to 7.5 th day of estrus, and the recipient cattle with insufficient corpus luteum firmness at the time of transfer are injected with 100 units of progesterone or 25 micrograms of luteinizing hormone. This estrus regimen is believed to have an estrus greater than 90%.
CN107372349A (application No. 201710791995.1) relates to a cross breeding method of a cool red bull and a jungle bull, wherein the jungle red bull is taken as a male parent and the cool red bull is taken as a female parent; after the heat of the cold red cow is confirmed, the cold red cow is frozen with the frozen semen of the bull, vitamin ADE is injected before freezing, and luteinizing hormone releasing hormone A3 is injected during freezing; performing concentrated feed supplement regulation and control according to body conditions in the perinatal period of the cold red cow, and supplementing calcium and magnesium preparations; the calf is separated from cow immediately after birth, and the calf diarrhea prevention bag is continuously taken for one week after the calf is fed with clean colostrum. The bred plain red bull and the bred cattle interspecific hybrid F1 generation have strong stress resistance, good fattening production performance and strong intramuscular fat deposition capability, and provide scientific basis for further improvement, reasonable utilization of resources and superior commercial value of the hybrid F1 generation of local fine cattle in China.
CN109964878A (application No. 201910384234.3) relates to a cross breeding method of Xinjiang Kazakh and Angus cattle, wherein the Angus cattle are used as male parents and the Xinjiang Kazakh are used as female parents; after the estrus of the cow of the Kazakh cattle in Xinjiang is confirmed, the cow is frozen with frozen semen of the bull of the Angus cattle, vitamin ADE is injected before freezing, and luteinizing hormone releasing hormone A3 is injected during freezing; carrying out concentrated feed supplement regulation and control according to body conditions and supplementing vitamin A and calcium-magnesium preparations in the perinatal period of the Xinjiang Kazakh cow; the calf is separated from cow immediately after birth, and the calf diarrhea prevention bag is continuously taken for one week after the calf is fed with clean colostrum. The hybrid between Xinjiang Kazakh cattle and Angus cattle bred by the method has strong stress resistance of F1 generation, has good fattening production performance, and provides scientific basis for further improvement of local improved Xinjiang Kazakh cattle in China, reasonable utilization of resources and commercial value of hybrid F1 generation superior medium-grade beef.
However, new methods for increasing estrus, ovulation, and conception in animals such as cattle are still desired in the art.
Disclosure of Invention
The object of the present invention is to provide a new method for increasing oestrus, ovulation and conception in animals such as cattle. It has been surprisingly found that the injection of luteinizing hormone releasing hormone a3 to achieve estrus synchronization in cattle and thus ovulation and conception in test animals provides unexpected results and the present invention has been completed based on such findings.
The invention makes the cow realize synchronous estrus by injecting the luteinizing hormone releasing hormone A3 into the cow, so as to ensure that the test animal ovulates and receives the fetus, and essentially, the invention is a method for improving the availability of the receptor cow in the cow embryo transplantation.
To this end, the present invention provides in a first aspect a method for increasing the availability of a recipient cow in a cow embryo transfer, comprising the steps of:
(1) providing a donor bovine: the donor cattle are nonpregnant and non-estrualized multiparous cows, and the anogenital tract diseases are confirmed through rectal examination; during the whole test period, grazing is mainly performed in the daytime, and forage is supplemented at night;
(2) administration: intramuscular injection of luteinizing hormone releasing hormone a3 in donor cattle;
(3) estrus and insemination: continuously observing the oestrus and ovulation conditions of the cows within 1-3 d from 0d on the day of medicine injection, recording the oestrus time and the oestrus ending time of the cows, observing the oestrus time and the oestrus ending time once every morning and evening for 2-3 h each time, determining the oestrus time, the oestrus program and the ovulation condition according to the appearance oestrus symptoms of the cows, determining oestrus synchronization according to rectal examination, carrying out artificial insemination on the oestrus and ovulation cows according to a freeze-matching technical operation rule, wherein the semen is the frozen semen of the same donor cow;
(4) and (3) pregnancy judgment: and (5) determining whether the cow is pregnant according to the rectal examination result 90 days after mating, calculating the estrus conception rate, and feeding the cow in the pregnancy according to a conventional method.
The method according to the first aspect of the present invention, wherein the donor bovine is selected from: and cattle, yellow cattle, cold red cattle, Kazakh cattle, and Angus cattle.
The method according to the first aspect of the present invention, wherein the donor cattle are cattle that are 4 months or more postpartum.
The method according to the first aspect of the present invention, wherein the donor cattle are cattle aged 3 to 8 years old.
The method according to the first aspect of the present invention, wherein the dose of the luteinizing hormone-releasing hormone A3 is 1 to 20 μ g/100kg body weight.
The method according to the first aspect of the present invention, wherein the dose of the luteinizing hormone-releasing hormone A3 is 2-10 μ g/100kg body weight.
The method according to the first aspect of the invention, wherein the dose of luteinizing hormone-releasing hormone A3 is 5 μ g/100kg body weight.
The method according to the first aspect of the invention, wherein said luteinizing hormone-releasing hormone a3 is administered by injection in a composition thereof.
The method according to the first aspect of the present invention wherein the composition comprises luteinizing hormone releasing hormone a3 and arginine hydrochloride, triethyl glycerol.
The method according to the first aspect of the present invention, wherein said composition comprises luteinizing hormone releasing hormone a3 and arginine hydrochloride, triethyl glycerol in a weight ratio of 5 μ g: 10-200 mg: 1-20 mg.
The method according to the first aspect of the present invention, wherein said composition comprises luteinizing hormone-releasing hormone a3 and arginine hydrochloride, triethyl glycerol in a weight ratio of 5 μ g: 20-100 mg: 2-10 mg.
The method according to the first aspect of the present invention, wherein said composition comprises luteinizing hormone-releasing hormone a3 and arginine hydrochloride, triethyl glycerol in a weight ratio of 5 μ g: 50 mg: 5 mg.
The method according to the first aspect of the present invention, wherein said composition is administered by injection after dissolution in water.
The method according to the first aspect of the present invention, wherein the composition is a freeze-dried powder injection.
The process according to the first aspect of the present invention, wherein the composition is prepared according to a process comprising the steps of: dissolving luteinizing hormone releasing hormone A3, arginine hydrochloride and triethyl glycerol in appropriate amount of water for injection (such as LHRH-A3 concentration of 1-20 μ g/ml, such as 2-10 μ g/ml, such as 5 μ g/ml), filtering with 0.22 μm microporous membrane for sterilization, packaging into penicillin bottles, and freeze drying.
The method according to the first aspect of the present invention, wherein the composition is administered by injection, diluted with water for injection to a concentration of LHRH-A3 of 1 to 20 μ g/ml, such as 2 to 10 μ g/ml, such as 5 μ g/ml, and administered by injection in a prescribed dose.
Further, the second aspect of the present invention provides a composition comprising luteinizing hormone releasing hormone a3, arginine hydrochloride, and triethyl glycerol.
The composition according to the second aspect of the invention comprises the luteinizing hormone releasing hormone A3, arginine hydrochloride and triethyl glycerol in a weight ratio of 5 mug: 10-200 mg: 1-20 mg.
The composition according to the second aspect of the invention comprises the luteinizing hormone releasing hormone A3, arginine hydrochloride and triethyl glycerol in a weight ratio of 5 mug: 20-100 mg: 2-10 mg.
The composition according to the second aspect of the invention comprises the luteinizing hormone releasing hormone A3, arginine hydrochloride and triethyl glycerol in a weight ratio of 5 mug: 50 mg: 5 mg.
The composition according to the second aspect of the present invention is administered by injection after dissolution in water.
The composition according to the second aspect of the present invention is a freeze-dried powder injection.
A composition according to the second aspect of the present invention, which is prepared according to a process comprising the steps of: dissolving luteinizing hormone releasing hormone A3, arginine hydrochloride and triethyl glycerol in appropriate amount of water for injection (such as LHRH-A3 concentration of 1-20 μ g/ml, such as 2-10 μ g/ml, such as 5 μ g/ml), filtering with 0.22 μm microporous membrane for sterilization, packaging into penicillin bottles, and freeze drying.
The composition according to the second aspect of the present invention is diluted with water for injection to a concentration of LHRH-A3 of 1 to 20. mu.g/ml, for example 2 to 10. mu.g/ml, for example 5. mu.g/ml, and is administered by injection in a prescribed dose.
Further, the third aspect of the present invention provides the use of luteinizing hormone releasing hormone A3 in the preparation of a pharmaceutical composition for increasing the availability of a recipient cow in bovine embryo transfer.
A method according to the third aspect of the invention, wherein the composition is as described in any embodiment of the second aspect of the invention.
The method according to the third aspect of the invention, wherein the method for increasing the availability of recipient cattle in a bovine embryo transfer is as described in any one of the embodiments of the first aspect of the invention.
Any technical feature possessed by any one aspect of the invention or any embodiment of that aspect is equally applicable to any other embodiment or any embodiment of any other aspect, so long as they are not mutually inconsistent, although appropriate modifications to the respective features may be made as necessary when applicable to each other. Various aspects and features of the disclosure are described further below.
All documents cited herein are incorporated by reference in their entirety and to the extent such documents do not conform to the meaning of the present invention, the present invention shall control. Further, the various terms and phrases used herein have the ordinary meaning as is known to those skilled in the art, and even though such terms and phrases are intended to be described or explained in greater detail herein, reference is made to the term and phrase as being inconsistent with the known meaning and meaning as is accorded to such meaning throughout this disclosure.
In the present invention, the luteinizing hormone-releasing hormone a3, english name: luteinizing Hormone Releasing Hormone A3, also called ovulation promoting NO. 3, ovulation promoting NO. three, ovulation promoting Hormone NO. three, polyembryon, etc., abbreviated as LHRH-A3 in English.
Luteinizing hormone releasing hormone A3(LHRH-A3) as a hormone medicine can promote the release of Luteinizing Hormone (LH) and Follicle Stimulating Hormone (FSH) from anterior pituitary of animals to promote ovarian follicle maturation and ovulation, so that the pituitary can be immediately released as synthesized hormone, and hormone synthesis can be stimulated. Used for treating cow ovulation retardation, ovary quiescence, corpus luteum persistence, ovarian cyst and early pregnancy diagnosis: can also be used for inducing ovulation in fish. For male animals, spermatogenesis can be promoted.
Luteinizing hormone releasing hormone a3 is commonly used in the field of animal husbandry: 1. increasing the number born. 2. Improve the estrus conception rate. 3. Treating follicular maturity difference, ovulation delay, and follicular cyst. 4. Enhance the effect of superovulation. 5. Strengthen the function of corpus luteum. 6. Can be used for inducing ovulation in fish.
The application of the luteinizing hormone releasing hormone A3 in the breeding of cattle is mainly as follows:
promoting the restoration of ovarian function, shortening calving interval, and carrying out intramuscular injection of 5-15 microgram within 8-18 days after delivery for 1 time every day for 3 days;
and (3) the tire receiving rate is improved: a. when in mating, 25 mu g is injected intramuscularly, which can obviously improve the estrus conception rate; b. intramuscular injection of 25 mu g for about 40 days after delivery, 0.4-0.6 mg of alprostadil sodium (carbendazim) for 6 days and 20 mu g for 1 day, which can obviously improve the conception rate within 60 days after delivery;
treating follicular dysplasia and ovulation retardation: 25-30 mu g of intramuscular injection is carried out during or 2 hours before the hybridization;
treatment of multiple follicular development: intramuscular injection of 25 μ g per day for 3 days;
treating ovarian cyst: intramuscular injection of 25 μ g per day for 5 consecutive days, and 100mg of progesterone per day, 3 months after pregnancy should be administered multiple times to prevent recurrence.
The application of the luteinizing hormone releasing hormone A3 in pig breeding mainly comprises the following steps:
the conception rate and the litter size of the sows in the estrus are improved: the sows are injected with 25-50 mu g of intramuscular in the afternoon of the 2 nd day of estrus, and are bred for 1 time in the morning and afternoon of the 3 rd day, and the average litter size of the sows is increased by 1.5-2;
the boar has weak libido, and 25 mu g of boar is taken every day for 3-5 days continuously.
The application of luteinizing hormone releasing hormone A3 in sheep reproduction mainly comprises the following steps:
after the ewes normally estre, 5-10 mu g of the goat feed is injected into the ewes by muscle injection, and after 4-6 hours, hybridization is carried out (if the ewes do not estre, 0.5-1 ml of three-in-one hormone can be used for promoting estrus first), so that the twins rate of the goats is obviously increased, and the number of the sheep born is increased.
The application of the luteinizing hormone releasing hormone A3 in other domestic animals is mainly as follows:
increasing conception rate and litter size, and intramuscular injection at the same time of or several hours before mating: deer 15 microgram/head, rabbit 0.2-0.5 microgram/head, dog fox 15 microgram/head, and hide animal 5 microgram/head.
The application of the luteinizing hormone releasing hormone A3 in aquatic products mainly comprises the following steps: ovulation promotion: intraperitoneal injection is carried out, wherein each fish is administrated once, the amount of grass carp is 2-5 mu g, and the amount of silver carp and bighead carp is 3-5 mu g.
The invention has surprisingly found that the effects of estrus, ovulation and conception of animals in the same period can be significantly improved by using the method of the invention.
Detailed Description
The present invention will be further described by the following examples, however, the scope of the present invention is not limited to the following examples. It will be understood by those skilled in the art that various changes and modifications may be made to the invention without departing from the spirit and scope of the invention. The present invention has been described generally and/or specifically with respect to materials used in testing and testing methods. Although many materials and methods of operation are known in the art for the purposes of this invention, the invention is nevertheless described herein in as detail as possible. Unless otherwise specified, the technical means used in the examples are conventional means well known to those skilled in the art, and the raw materials used are commercially available products.
Example 1: test of estrus synchronization
The estrus synchronization effect test was performed as follows.
(1) Providing a donor bovine: the method comprises the following steps of (1) confirming the diseases of the genital tract by rectal examination of south-yang yellow cattle, which are born for more than 4 months, aged 3-8 years, barren and unhappy multiparous cows; during the whole test period, grazing is mainly performed in the daytime, and forage is supplemented at night;
(2) administration: intramuscular injection of luteinizing hormone-releasing hormone A3(LHRH-A3) to donor cattle at a dose of 5 μ g/100kg body weight;
(3) estrus and insemination: continuously observing the oestrus and ovulation conditions of the cows within 1-3 d from 0d on the day of medicine injection, recording the oestrus time and the oestrus ending time of the cows, observing the oestrus once in the morning and at night every day for 2-3 h each time, determining the oestrus time, the oestrus program and the ovulation condition according to the external oestrus symptoms of the cows, determining oestrus synchronization according to rectal examination, performing artificial insemination on the oestrus and ovulation cows according to a freeze-matching technical operation rule at a proper time, wherein the semen is the frozen semen of the Nanyang yellow cattle;
(4) and (3) pregnancy judgment: and (5) determining whether the cow is pregnant or not according to the rectal examination result 90d after the mating, calculating the estrus conception rate, and feeding the cow in the pregnancy according to a conventional method.
In this example, the preparation of the drugs used in step (2): dissolving commercially available raw material medicine (> 99%) of luteinizing hormone releasing hormone A3 with water for injection (5 μ g/ml), filtering with 0.22 μm microporous membrane for sterilization, subpackaging in penicillin bottles, and freeze-drying for use; when in use, the medicine is diluted to 5 mu g/ml by water for injection and is injected according to the regulated dosage.
The results of some embodiments of the present application are as follows:
note: in the table, ovulatory animals are counted on the basis of estrus in the same period, estrus pregnancies are counted on the basis of ovulatory animals, and total pregnancies are counted on the basis of the total number of test animals. It has been surprisingly found that the injection of both arginine hydrochloride and triethyl glycerol together with the injection of luteinizing hormone-releasing hormone a3 according to the present invention can significantly increase conception rates.
Example 2: test of estrus synchronization
The estrus synchronization effect test was performed as follows.
(1) Providing a donor bovine: the method comprises the following steps of (1) confirming the diseases of the genital tract by rectal examination of south-yang yellow cattle, which are born for more than 4 months, aged 3-8 years, barren and unhappy multiparous cows; during the whole test period, grazing is mainly performed in the daytime, and forage is supplemented at night;
(2) administration: intramuscular injection of a composition comprising luteinizing hormone releasing hormone A3(LHRH-A3) to donor cattle at a dose of 5 μ g per 100kg body weight calculated as LHRH-A3;
(3) estrus and insemination: continuously observing the oestrus and ovulation conditions of the cows within 1-3 d from 0d on the day of medicine injection, recording the oestrus time and the oestrus ending time of the cows, observing the oestrus once in the morning and at night every day for 2-3 h each time, determining the oestrus time, the oestrus program and the ovulation condition according to the external oestrus symptoms of the cows, determining oestrus synchronization according to rectal examination, performing artificial insemination on the oestrus and ovulation cows according to a freeze-matching technical operation rule at a proper time, wherein the semen is the frozen semen of the Nanyang yellow cattle;
(4) and (3) pregnancy judgment: and (5) determining whether the cow is pregnant according to the rectal examination result 90 days after mating, calculating the estrus conception rate, and feeding the cow in the pregnancy according to a conventional method.
The composition administered in step (2) of this example contains LHRH-a3, arginine hydrochloride, and triethyl glycerol in a weight ratio of 5 μ g: 50 mg: 5mg, the preparation method of the composition comprises the following steps: dissolving commercially available bulk drug of luteinizing hormone releasing hormone A3 (> 99%) and arginine hydrochloride and triethyl glycerolate in proportion with water for injection (to LHRH-A3 concentration of 5 μ g/ml), filtering with 0.22 μm microporous membrane for sterilization, subpackaging in penicillin bottles, and freeze-drying for use; when in use, the injection is diluted by water for injection until the concentration of LHRH-A3 is 5 mug/ml, and the injection is administered according to the regulated dose.
Example 3: test of estrus synchronization
The estrus synchronization effect test was performed as follows.
(1) Providing a donor bovine: the method comprises the following steps of (1) confirming the diseases of the genital tract by rectal examination of south-yang yellow cattle, which are born for more than 4 months, aged 3-8 years, barren and unhappy multiparous cows; during the whole test period, grazing is mainly performed in the daytime, and forage is supplemented at night;
(2) administration: intramuscular injection of a composition comprising luteinizing hormone releasing hormone A3(LHRH-A3) to donor cattle at a dose of 5 μ g per 100kg body weight calculated as LHRH-A3;
(3) estrus and insemination: continuously observing the oestrus and ovulation conditions of the cows within 1-3 d from 0d on the day of medicine injection, recording the oestrus time and the oestrus ending time of the cows, observing the oestrus once in the morning and at night every day for 2-3 h each time, determining the oestrus time, the oestrus program and the ovulation condition according to the external oestrus symptoms of the cows, determining oestrus synchronization according to rectal examination, performing artificial insemination on the oestrus and ovulation cows according to a freeze-matching technical operation rule at a proper time, wherein the semen is the frozen semen of the Nanyang yellow cattle;
(4) and (3) pregnancy judgment: and (5) determining whether the cow is pregnant according to the rectal examination result 90 days after mating, calculating the estrus conception rate, and feeding the cow in the pregnancy according to a conventional method.
The composition administered in step (2) of this example contains LHRH-a3 and arginine hydrochloride in a weight ratio of 5 μ g: 50mg, the preparation method of the composition comprises the following steps: dissolving commercially available bulk drug (> 99%) of luteinizing hormone releasing hormone A3 and arginine hydrochloride in water for injection according to a certain proportion (the concentration of LHRH-A3 is 5 mu g/ml), filtering and sterilizing by using a 0.22 mu m microporous filter membrane, subpackaging into a penicillin bottle, and freeze-drying for later use; when in use, the injection is diluted by water for injection until the concentration of LHRH-A3 is 5 mug/ml, and the injection is administered according to the regulated dose.
Example 4: test of estrus synchronization
The estrus synchronization effect test was performed as follows.
(1) Providing a donor bovine: the method comprises the following steps of (1) confirming the diseases of the genital tract by rectal examination of south-yang yellow cattle, which are born for more than 4 months, aged 3-8 years, barren and unhappy multiparous cows; during the whole test period, grazing is mainly performed in the daytime, and forage is supplemented at night;
(2) administration: intramuscular injection of a composition comprising luteinizing hormone releasing hormone A3(LHRH-A3) to donor cattle at a dose of 5 μ g per 100kg body weight calculated as LHRH-A3;
(3) estrus and insemination: continuously observing the oestrus and ovulation conditions of the cows within 1-3 d from 0d on the day of medicine injection, recording the oestrus time and the oestrus ending time of the cows, observing the oestrus once in the morning and at night every day for 2-3 h each time, determining the oestrus time, the oestrus program and the ovulation condition according to the external oestrus symptoms of the cows, determining oestrus synchronization according to rectal examination, performing artificial insemination on the oestrus and ovulation cows according to a freeze-matching technical operation rule at a proper time, wherein the semen is the frozen semen of the Nanyang yellow cattle;
(4) and (3) pregnancy judgment: and (5) determining whether the cow is pregnant according to the rectal examination result 90 days after mating, calculating the estrus conception rate, and feeding the cow in the pregnancy according to a conventional method.
The composition administered in step (2) of this example contained LHRH-a3 in a weight ratio of 5 μ g to triethyl glycerol: 5mg, the preparation method of the composition comprises the following steps: dissolving commercially available bulk drug (> 99%) of luteinizing hormone releasing hormone A3 and triethyl glycerolate in proportion with water for injection (to LHRH-A3 concentration of 5 μ g/ml), filtering with 0.22 μm microporous membrane for sterilization, packaging into penicillin bottles, and freeze drying; when in use, the injection is diluted by water for injection until the concentration of LHRH-A3 is 5 mug/ml, and the injection is administered according to the regulated dose.
Example 5:test of estrus synchronization
The estrus synchronization effect test was performed on 41 Japanese pure blood and cattle according to the method of example 2, and the results showed that: the estrus count is 38 (the synchronization rate is 92.7%), the ovulation count is 35 (the ovulation rate is 92.1%), the conception count is 31 (the estrus conception rate is 88.6%), and the total conception rate is 75.6%.
Although the invention has been described in detail hereinabove with respect to a general description and specific embodiments thereof, it will be apparent to those skilled in the art that modifications or improvements may be made thereto based on the invention. Accordingly, such modifications and improvements are intended to be within the scope of the invention as claimed.
Claims (13)
1. The method for improving the availability of the receptor cattle in the cattle embryo transplantation comprises the following steps:
(1) cattle 4 months or more post-partum were provided as donor cattle: the donor cattle are nonpregnant and non-estrualized multiparous cows, and the anogenital tract diseases are confirmed through rectal examination; during the whole test period, grazing is mainly performed in the daytime, and forage is supplemented at night;
(2) administration: intramuscular injection of a composition comprising luteinizing hormone-releasing hormone a3 and arginine hydrochloride and triethyl glycerolate in a weight ratio of 5 μ g: 20-100 mg: 2-10 mg of luteinizing hormone releasing hormone A3, wherein the dosage of the luteinizing hormone releasing hormone A3 is 2-10 mug/100 kg of body weight;
(3) estrus and insemination: continuously observing the oestrus and ovulation conditions of the cows within 1-3 d from 0d on the day of medicine injection, recording the oestrus time and the oestrus ending time of the cows, observing the oestrus time and the oestrus ending time once every morning and evening for 2-3 h each time, determining the oestrus time, the oestrus program and the ovulation condition according to the appearance oestrus symptoms of the cows, determining oestrus synchronization according to rectal examination, carrying out artificial insemination on the oestrus and ovulation cows according to a freeze-matching technical operation rule, wherein the semen is the frozen semen of the same donor cow;
(4) and (3) pregnancy judgment: and (5) determining whether the cow is pregnant according to the rectal examination result 90 days after mating, calculating the estrus conception rate, and feeding the cow in the pregnancy according to a conventional method.
2. The method of claim 1, wherein the donor bovine is selected from the group consisting of: and cattle, yellow cattle, cold red cattle, Kazakh cattle, and Angus cattle.
3. The method of claim 1, wherein the donor cattle are cattle aged 3-8 years.
4. The method of claim 1, wherein the dose of luteinizing hormone releasing hormone a3 is 5 μ g/100kg body weight.
5. The method according to claim 1, wherein the weight ratio of luteinizing hormone releasing hormone a3, arginine hydrochloride, and triethyl glycerol in the composition is 5 μ g: 50 mg: 5 mg.
6. The method of claim 1, wherein the composition is administered by injection after dissolution in water.
7. The method of claim 1, wherein the composition is a lyophilized powder injection.
8. The method of claim 1, wherein the composition is prepared according to a process comprising the steps of: dissolving luteinizing hormone releasing hormone A3, arginine hydrochloride and triethyl glycerol in a proper amount of water for injection according to a proportion until the concentration of LHRH-A3 is 2-10 mug/ml, filtering and sterilizing by using a 0.22 mug microporous filter membrane, subpackaging in a penicillin bottle, and freeze-drying to obtain the luteinizing hormone releasing hormone.
9. The method according to claim 1, wherein the composition is administered by injection, diluted with water for injection to an LHRH-A3 concentration of 5 μ g/ml, and administered by injection at a prescribed dose.
10. A composition for use in a method for increasing the availability of a recipient cow in a bovine embryo transfer as claimed in any one of claims 1 to 9, wherein the composition comprises luteinizing hormone releasing hormone a3, arginine hydrochloride, and triethyl glycerol in a weight ratio of 5 μ g: 20-100 mg: 2-10 mg.
11. The composition according to claim 10, wherein the weight ratio of the luteinizing hormone releasing hormone a3, arginine hydrochloride, and triethyl glycerol is 5 μ g: 50 mg: 5 mg.
12. The composition of claim 10, which is a lyophilized powder for injection prepared according to a method comprising the steps of: dissolving luteinizing hormone releasing hormone A3, arginine hydrochloride and triethyl glycerol in a proper amount of water for injection according to a proportion until the concentration of LHRH-A3 is 2-10 mug/ml, filtering and sterilizing by using a 0.22 mug microporous filter membrane, subpackaging in a penicillin bottle, and freeze-drying to obtain the luteinizing hormone releasing hormone.
13. Use of luteinizing hormone releasing hormone a3 for the manufacture of a pharmaceutical composition for increasing the availability of recipient cattle in bovine embryo transfer, said composition being as claimed in any one of claims 10 to 12.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202010467616.5A CN111568600B (en) | 2020-05-28 | 2020-05-28 | Method for improving availability of acceptor cattle in cattle embryo transplantation |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202010467616.5A CN111568600B (en) | 2020-05-28 | 2020-05-28 | Method for improving availability of acceptor cattle in cattle embryo transplantation |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN111568600A CN111568600A (en) | 2020-08-25 |
| CN111568600B true CN111568600B (en) | 2022-05-10 |
Family
ID=72125542
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN202010467616.5A Active CN111568600B (en) | 2020-05-28 | 2020-05-28 | Method for improving availability of acceptor cattle in cattle embryo transplantation |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN111568600B (en) |
Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101066046A (en) * | 2007-01-19 | 2007-11-07 | 浙江省淡水水产研究所 | Curled-lip mandarin fish and spot and spotted mandarin fish hybridizing propagation process |
| CN105342723A (en) * | 2015-12-03 | 2016-02-24 | 陈兴 | Method for increasing conception rate of artificial insemination of milk buffaloes |
| CN105638568A (en) * | 2016-03-03 | 2016-06-08 | 曲靖创亿农牧有限公司 | Beef cattle breeding method |
| CN106614369A (en) * | 2016-12-29 | 2017-05-10 | 刘桂 | Breeding method for promoting pig breeding |
| CN106719312A (en) * | 2015-11-25 | 2017-05-31 | 陈兴 | A kind of method that dairy buffalo cultivates artificial insemination |
-
2020
- 2020-05-28 CN CN202010467616.5A patent/CN111568600B/en active Active
Patent Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101066046A (en) * | 2007-01-19 | 2007-11-07 | 浙江省淡水水产研究所 | Curled-lip mandarin fish and spot and spotted mandarin fish hybridizing propagation process |
| CN106719312A (en) * | 2015-11-25 | 2017-05-31 | 陈兴 | A kind of method that dairy buffalo cultivates artificial insemination |
| CN105342723A (en) * | 2015-12-03 | 2016-02-24 | 陈兴 | Method for increasing conception rate of artificial insemination of milk buffaloes |
| CN105638568A (en) * | 2016-03-03 | 2016-06-08 | 曲靖创亿农牧有限公司 | Beef cattle breeding method |
| CN106614369A (en) * | 2016-12-29 | 2017-05-10 | 刘桂 | Breeding method for promoting pig breeding |
Non-Patent Citations (1)
| Title |
|---|
| 浅谈生殖激素在奶牛繁殖中的应用;刘文国;《中国奶牛》;19970910(第05期);第31-32页 * |
Also Published As
| Publication number | Publication date |
|---|---|
| CN111568600A (en) | 2020-08-25 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| US9763962B2 (en) | Estrus synchronization preparations and effective CIDR-less protocols | |
| Boland et al. | Alternative gonadotrophins for superovulation in cattle | |
| Rathbone et al. | Recent advances in bovine reproductive endocrinology and physiology and their impact on drug delivery system design for the control of the estrous cycle in cattle | |
| Islam | Synchronization of estrus in cattle: a review. | |
| Zimbelman | Determination of the Minimal Effective Dose of 6 a-Methyl-17 a-Acetoxyprogesterone for Control of the Estrual Cycle of Cattle | |
| Ryan et al. | Pregnancy rates in dairy cows following the administration of a GnRH analogue at the time of artificial insemination or at mid-cycle post insemination | |
| CN111588834B (en) | Method for improving bovine embryo transplantation estrus synchronization effect and used medicine | |
| CN111568600B (en) | Method for improving availability of acceptor cattle in cattle embryo transplantation | |
| Lopez-Gatius | Ovarian response to prostaglandin F2α in lactating dairy cows: A clinical update | |
| Chu | The influence of the thyroid on pregnancy and parturition in the rabbit | |
| Pressing | Pharmacologic control of swine reproduction | |
| Çizmeci et al. | Superovulation in cows: a review. | |
| Day | Induced termination of the calving season in a large dairy herd | |
| Larson et al. | The bovine estrous cycle and synchronization of estrus | |
| RU2750208C1 (en) | Method for obtaining embryos from heifers-donors of holstein breed | |
| Johnson et al. | Gonadotropin induced multiple births in beef cows following estrus synchronization | |
| Nergiz et al. | The Effect of PMSG Treatment on Pregnancy Rates in Synchronized Heifers using Norgestomet Ear Implants | |
| RU2808263C1 (en) | Method of tightening timing of introducing heifers into reproduction | |
| Gordon | The use of progesterone and serum gonadotrophin (PMS) in the control of fertility in sheep. II. Studies in the extra-seasonal production of lambs | |
| CN117298256A (en) | Gonadotrophin composition for improving reproduction performance of yaks and application method thereof | |
| Ola et al. | Oestrus synchronisation with progestagen injectables in West African Dwarf does. | |
| Beal | Estrous synchronization of cyclic and anestrous cows with Syncro-Mate-B® | |
| Al-Sharari et al. | COMPARISON BETWEEN SHORT-AND LONG-TERM OF ESTRUS SYNCHRONIZATION WITH DIFFERENT DOSES OF PMSG IN NOEMI EWES. | |
| Norwood | Factors affecting postpartum regression of the bovine uterus | |
| Oxender et al. | Some potential uses of prostaglandins in domestic animals |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| PB01 | Publication | ||
| PB01 | Publication | ||
| SE01 | Entry into force of request for substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| CB02 | Change of applicant information | ||
| CB02 | Change of applicant information |
Address after: 300457 building a, Alexandria, No.3 Haitong street, Binhai New Area Development Zone, Tianjin Boya company Applicant after: Tianjin Limu Biotechnology Co.,Ltd. Address before: 300457 building a, Alexandria, No.3 Haitong street, Binhai New Area Development Zone, Tianjin Boya company Applicant before: TIANJIN BOYU LIMU TECHNOLOGY CO.,LTD. |
|
| GR01 | Patent grant | ||
| GR01 | Patent grant |