CN115153947A - Method for superovulation of Argwang sheep - Google Patents
Method for superovulation of Argwang sheep Download PDFInfo
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- CN115153947A CN115153947A CN202210810428.7A CN202210810428A CN115153947A CN 115153947 A CN115153947 A CN 115153947A CN 202210810428 A CN202210810428 A CN 202210810428A CN 115153947 A CN115153947 A CN 115153947A
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- 241001494479 Pecora Species 0.000 title claims abstract description 44
- 238000000034 method Methods 0.000 title claims abstract description 28
- 206010042573 Superovulation Diseases 0.000 title claims abstract description 20
- 102000012673 Follicle Stimulating Hormone Human genes 0.000 claims abstract description 34
- 108010079345 Follicle Stimulating Hormone Proteins 0.000 claims abstract description 34
- 229940028334 follicle stimulating hormone Drugs 0.000 claims abstract description 26
- 210000001161 mammalian embryo Anatomy 0.000 claims abstract description 24
- RJKFOVLPORLFTN-LEKSSAKUSA-N Progesterone Chemical compound C1CC2=CC(=O)CC[C@]2(C)[C@@H]2[C@@H]1[C@@H]1CC[C@H](C(=O)C)[C@@]1(C)CC2 RJKFOVLPORLFTN-LEKSSAKUSA-N 0.000 claims abstract description 23
- 244000125300 Argania sideroxylon Species 0.000 claims abstract description 17
- 238000002347 injection Methods 0.000 claims abstract description 11
- 239000007924 injection Substances 0.000 claims abstract description 11
- 239000000186 progesterone Substances 0.000 claims abstract description 11
- 229960003387 progesterone Drugs 0.000 claims abstract description 11
- 239000000829 suppository Substances 0.000 claims abstract description 10
- 238000011010 flushing procedure Methods 0.000 claims abstract description 9
- 230000013011 mating Effects 0.000 claims abstract description 9
- 239000011782 vitamin Substances 0.000 claims abstract description 7
- 229940088594 vitamin Drugs 0.000 claims abstract description 7
- 229930003231 vitamin Natural products 0.000 claims abstract description 7
- 235000013343 vitamin Nutrition 0.000 claims abstract description 7
- 150000003722 vitamin derivatives Chemical class 0.000 claims abstract description 7
- 230000016087 ovulation Effects 0.000 claims abstract description 5
- 210000001215 vagina Anatomy 0.000 claims abstract description 5
- 238000011084 recovery Methods 0.000 claims description 10
- 238000005406 washing Methods 0.000 claims description 7
- AXXJUGQWKHTJBS-OALUTQOASA-N ClC(=CCCCCC[C@H]1CCC[C@@H]1CCCCCCCC)O Chemical compound ClC(=CCCCCC[C@H]1CCC[C@@H]1CCCCCCCC)O AXXJUGQWKHTJBS-OALUTQOASA-N 0.000 claims description 5
- 230000003444 anaesthetic effect Effects 0.000 claims description 4
- 238000010255 intramuscular injection Methods 0.000 abstract description 4
- 239000007927 intramuscular injection Substances 0.000 abstract description 4
- 230000003631 expected effect Effects 0.000 abstract description 3
- 239000000203 mixture Substances 0.000 abstract description 3
- 230000006698 induction Effects 0.000 abstract 1
- 239000007788 liquid Substances 0.000 description 6
- 238000005516 engineering process Methods 0.000 description 3
- 238000004519 manufacturing process Methods 0.000 description 3
- 206010002091 Anaesthesia Diseases 0.000 description 2
- 230000037005 anaesthesia Effects 0.000 description 2
- 210000004246 corpus luteum Anatomy 0.000 description 2
- 244000144972 livestock Species 0.000 description 2
- 210000003101 oviduct Anatomy 0.000 description 2
- 102000006771 Gonadotropins Human genes 0.000 description 1
- 108010086677 Gonadotropins Proteins 0.000 description 1
- 241000282849 Ruminantia Species 0.000 description 1
- QVGXLLKOCUKJST-UHFFFAOYSA-N atomic oxygen Chemical compound [O] QVGXLLKOCUKJST-UHFFFAOYSA-N 0.000 description 1
- 210000002459 blastocyst Anatomy 0.000 description 1
- 230000008859 change Effects 0.000 description 1
- 230000007547 defect Effects 0.000 description 1
- 238000011161 development Methods 0.000 description 1
- 238000010586 diagram Methods 0.000 description 1
- 238000007599 discharging Methods 0.000 description 1
- 230000005059 dormancy Effects 0.000 description 1
- 210000002257 embryonic structure Anatomy 0.000 description 1
- 230000007613 environmental effect Effects 0.000 description 1
- 230000003203 everyday effect Effects 0.000 description 1
- 230000004720 fertilization Effects 0.000 description 1
- 230000008014 freezing Effects 0.000 description 1
- 238000007710 freezing Methods 0.000 description 1
- 239000002622 gonadotropin Substances 0.000 description 1
- 238000000338 in vitro Methods 0.000 description 1
- 230000001939 inductive effect Effects 0.000 description 1
- 230000000968 intestinal effect Effects 0.000 description 1
- 238000007443 liposuction Methods 0.000 description 1
- 230000007774 longterm Effects 0.000 description 1
- 239000008267 milk Substances 0.000 description 1
- 210000004080 milk Anatomy 0.000 description 1
- 230000004048 modification Effects 0.000 description 1
- 238000012986 modification Methods 0.000 description 1
- 210000000472 morula Anatomy 0.000 description 1
- 210000003205 muscle Anatomy 0.000 description 1
- 210000002394 ovarian follicle Anatomy 0.000 description 1
- 229910052760 oxygen Inorganic materials 0.000 description 1
- 239000001301 oxygen Substances 0.000 description 1
- 230000008569 process Effects 0.000 description 1
- 238000011160 research Methods 0.000 description 1
- 239000000243 solution Substances 0.000 description 1
- 238000012360 testing method Methods 0.000 description 1
- 238000002054 transplantation Methods 0.000 description 1
- 230000035899 viability Effects 0.000 description 1
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- 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
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- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Veterinary Medicine (AREA)
- Transplantation (AREA)
- Reproductive Health (AREA)
- Engineering & Computer Science (AREA)
- Wood Science & Technology (AREA)
- Zoology (AREA)
- Animal Behavior & Ethology (AREA)
- General Health & Medical Sciences (AREA)
- Public Health (AREA)
- Medicines That Contain Protein Lipid Enzymes And Other Medicines (AREA)
Abstract
The invention provides a method for superovulation of Arwann sheep, which comprises the following steps: placing progesterone suppository in vagina of donor Argomasum alfa sheep, performing intramuscular injection of vitamin ADE mixture at 1d and 7d, performing intramuscular injection of follicle-stimulating hormone by a gradual decrement method at 12h interval every morning and afternoon from 11d, and continuously injecting for 4d; injecting the cloroxen while injecting the follicle-stimulating hormone of the 7 th needle, and taking out the progesterone suppository when injecting the follicle-stimulating hormone of the 8 th needle; injection of LHRH-A3 for ovulation induction at 15d, and pilot mating at 15d and 16 d; after the mating, donors were fasted for 48 hours at the 19 th afternoon of the schedule and surgical embryo flushing was performed at 21 d. The invention successfully applies the superovulation method to the argan sheep for the first time and achieves the expected effect of average level of other sheep varieties in China.
Description
Technical Field
The invention belongs to the field of embryos, and particularly relates to a method for superovulation of Argomorpha sheep.
Background
The superovulation technology is a method for inducing multiple ovarian follicles to develop and discharging multiple ova with fertilization capability by applying exogenous gonadotropin, is called superovulation for short, is combined with an embryo transplantation technology, can greatly improve the production and propagation capability of excellent breeders, and is widely applied to production and scientific research.
The Arwana sheep is a special livestock species of Qinghai-Tibet plateau and is one of precious local livestock species resources in China. The argan sheep has the characteristics of suitability for living in environmental regions with high altitude, low temperature, large day and night temperature difference, short growth period of pasture, low oxygen pressure and the like. As a result of long-term selection of the special ecological environment, the argan sheep has extremely strong viability and is resistant to rough cold. The argan sheep is produced in a high-cold grassland without modern industrial pollution and is a genuine 'green food'. Meanwhile, the sheep raising industry is also an economic foundation and a supporting column industry of a high and cold pasturing area, and the continuous development of the production of the sheep raising industry has very important significance for improving the living standard of herdsmen.
At present, no super-exclusion technology for the Arwana sheep exists; therefore, it is important to provide a method for superovulation in argan sheep.
Disclosure of Invention
The technical problem to be solved by the invention is to provide a method for superovulation in Argomasa sheep aiming at the defects of the prior art, the method successfully applies the superovulation method to the Algomasa sheep in Tibet Changdu for the first time, and the expected effect of the average level in China is achieved.
In order to solve the technical problems, the invention adopts the technical scheme that: a method for superovulation of Argomorpha sheep, comprising the steps of:
placing a progesterone suppository in the vagina of a donor argan sheep aged 3-5 years, wherein the date for placing the progesterone suppository is set as 1d; at 7d, vitamin ADE cocktail was injected intramuscularly; from 11d, injecting the follicle-stimulating hormone intramuscularly by a gradual decrement method at an interval of 12 hours every morning and afternoon, and continuously injecting for 4d; injecting 0.1mg of chloroprostenol simultaneously with the injection of the 7 th needle of follitropin, and removing the progesterone suppository when the 8 th needle of follitropin is injected; at day 15d, the LHRH-A3 of 25 μ Ag is injected for ovulation promotion, and a trial mating is performed on days 15 and 16; following mating, fasting was performed for 48h at 19d and surgical embryo flushing was performed at 21 d.
Preferably, the dosage of the vitamin ADE mixture is 3ml; the Arwann sheep is Tibet Changdu Arwann sheep.
Preferably, the weight of the donor argan sheep is more than or equal to 50kg, high-dose follicle-stimulating hormone is adopted, the total dosage of the high-dose follicle-stimulating hormone is 500IU, the high-dose follicle-stimulating hormone is diluted to 50IU/mL, and the injection doses are respectively 2.3 mL/time, 1.2 mL/time, 1.0 mL/time, 0.5 mL/time and 0.5 mL/time;
the weight of the donor argan sheep is 40-50 kg, low-dose follicle-stimulating hormone is adopted, the total dosage of the low-dose follicle-stimulating hormone is 375IU, the low-dose follicle-stimulating hormone is diluted to 50IU/mL, and the injection doses are 1.8 mL/time, 1.0 mL/time, 0.6 mL/time, 0.3 mL/time and 0.3 mL/time respectively.
Preferably, the dosage of the anesthesia for surgical embryo flushing is 0.5 ml-0.6 ml.
Preferably, the embryo recovery of the surgical embryo washing adopts a uterine horn recovery method.
Compared with the prior art, the invention has the following advantages:
the invention successfully applies the superovulation method to the Argomorpha sheep for the first time and achieves the expected effect of average level in China.
The present invention will be described in further detail with reference to the accompanying drawings and examples.
Drawings
FIG. 1 is a schematic flow diagram of the superarranging method of the present invention.
Detailed Description
The method for superovulation of argan sheep comprises the following steps:
placing progesterone suppository (CIDR) in vagina of donor Argomasus sheep aged from 3 to 5 years, wherein the date of placing progesterone suppository is set as 1d; at 7d, 3ml of vitamin ADE mixture is injected into the muscle; starting from 11d, injecting 7 am every day, and injecting Follicle Stimulating Hormone (FSH) intramuscularly by a gradual decrement method at an interval of 12h in the morning and afternoon, wherein 4d are continuously injected and 8 needles are injected in total; injecting 0.1mg of chloroprostenol (PG) simultaneously with injection of Follicle Stimulating Hormone (FSH) at the 7 th needle, and removing the progesterone plug at the time of injection of Follicle Stimulating Hormone (FSH) at the 8 th needle; at day 15d, the LHRH-A3 of 25 μ Ag is injected for ovulation promotion, and a trial mating is performed on days 15 and 16; after the mating, fasting is carried out for 48h in the afternoon of the 19d, and surgical embryo flushing is carried out at the 21 d; the donor Argomashes are Tibet Changdu Argomashes;
the weight of the Donor Argomashes argan sheep is more than or equal to 50kg, high-dose follicle-stimulating hormone is adopted, the total dosage of the high-dose follicle-stimulating hormone is 500IU, the high-dose follicle-stimulating hormone is diluted to 50IU/mL, and the injection doses are respectively 2.3 mL/time, 1.2 mL/time, 1.0 mL/time, 0.5 mL/time and 0.5 mL/time;
the weight of the donor argan sheep is 40-50 kg, low-dose follicle-stimulating hormone is adopted, the total dosage of the low-dose follicle-stimulating hormone is 375IU, the low-dose follicle-stimulating hormone is diluted to 50IU/mL, and the injection doses are 1.8 mL/time, 1.0 mL/time, 0.6 mL/time, 0.3 mL/time and 0.3 mL/time respectively;
in the embodiment, the dosage of the anesthesia for embryo flushing in the operation is 0.5 ml-0.6 ml; the weight of the donor Arwana sheep is lower than 50kg, and 0.5ml of anesthetic is used for neck intramuscular injection; the weight of the donor Arwana sheep is higher than 50kg, the dosage of the anesthetic is adjusted to 0.6ml, and the administration mode is neck intramuscular injection; the donor sheep is anesthetized by adopting a ruminant anesthetic commonly used in China, namely 'fast dormancy new' and is provided with 'revival' at the same time for emergency treatment when the sheep is overanesthetized;
in this example, the vitamin ADE cocktail was purchased from beijing & lttm & gt, specification 5ml by 10 per box; the follitropin is purchased from Ningbo Sansheng, specification 500IU 10 bottles/boxes; the chloroprostenol is purchased from Shanghai Ministry, and the specification is 2 mg/branch by 10 branches/box; the LHRH-A3 promotes ovulation and is purchased from Ningbo three-born, and the specification is 25ug x 10 pieces/box;
in the embodiment, the embryo recovery of the surgical embryo washing adopts a uterine horn recovery method to recover the embryo at the 6 th day after the mating, the embryo at the stage should develop to a compact morula or early blastocyst stage, the embryo at the stage normally enters the uterine horn, and the recovery operation is relatively easy; in addition, the adaptability of the embryo in the period to the in vitro operation is strong, and the embryo is convenient to carry out the freezing storage operation; embryo washing is to inject embryo washing liquid from the uterine horn tip to the direction of the uterine tube joint part by using a venous indwelling needle, the recovery of embryo washing liquid is to insert a uterine cavity in the middle of the uterine horn by using a three-cavity catheter (8 FR) special for children, and the tip of the catheter points to the uterine tube joint part; injecting liquid into the closed ball in the catheter to expand and block the uterine horn cavity, so as to prevent the flushing liquid from flowing to one side of the vagina of the uterine horn; a plurality of test results show that the uterine horn of the argan sheep has high brittleness, the inner wall of the uterine cavity is easily torn in the process of expanding the sealed ball, then a porous milk duct (or a fat-reducing special liposuction needle) with a rubber tube for connection is directly used for recovering the embryo flushing liquid, and meanwhile, the uterine cavity is sealed at the vaginal side of the uterine cavity inlet of the recovery tube by using intestinal forceps with the rubber tube, so that the embryo flushing liquid is ensured to completely return to the collection triangular bottle through the recovery tube and not to flow back to the uterine horn at the vaginal end.
After the method for superovulation is implemented, the average number of corpus luteum of the Xizang Changdu Argwang sheep is 10, and exceeds the average number of corpus luteum of the domestic superovulation method by 7-8.
The above description is only for the preferred embodiment of the present invention, and is not intended to limit the present invention in any way. Any simple modification, change and equivalent changes of the above embodiments according to the technical essence of the invention are still within the protection scope of the technical solution of the invention.
Claims (5)
1. A method for superovulation of Argwang sheep is characterized by comprising the following steps:
placing a progesterone suppository in the vagina of a donor argan sheep aged 3-5 years, wherein the date for placing the progesterone suppository is set as 1d; at 7d, vitamin ADE cocktail was injected intramuscularly; from 11d, injecting the follicle-stimulating hormone intramuscularly by a gradual decrement method at an interval of 12 hours every morning and afternoon, and continuously injecting for 4d; injecting 0.1mg of chloroprostenol simultaneously with the injection of the 7 th needle of follitropin, and removing the progesterone suppository when the 8 th needle of follitropin is injected; at day 15d, the LHRH-A3 of 25 μ Ag is injected for ovulation promotion, and a trial mating is performed on days 15 and 16; following mating, fasting was performed for 48h at 19d and surgical embryo flushing was performed at 21 d.
2. The method for superovulation in argan sheep as claimed in claim 1, wherein said vitamin ADE cocktail is administered in an amount of 3ml; the donor Arwana sheep is Tibet Changdu Arwana sheep.
3. The method for superovulation in Argomasa sheep as claimed in claim 1, wherein said donor Argomasa sheep weighs more than or equal to 50kg, and high dose of follitropin is used, wherein the total dosage of said high dose of follitropin is 500IU, and the high dose of follitropin is diluted to 50IU/mL, and the injection dose is 2.3 mL/time, 1.2 mL/time, 1.0 mL/time, 0.5 mL/time and 0.5 mL/time respectively;
the weight of the donor argan sheep is 40-50 kg, low-dose follicle-stimulating hormone is adopted, the total dosage of the low-dose follicle-stimulating hormone is 375IU, the low-dose follicle-stimulating hormone is diluted to 50IU/mL, and the injection doses are 1.8 mL/time, 1.0 mL/time, 0.6 mL/time, 0.3 mL/time and 0.3 mL/time respectively.
4. The method for superovulation in argan sheep as claimed in claim 1, wherein said surgical embryo washing is performed in an anesthetic dose of 0.5ml to 0.6ml.
5. The method for superovulation in an argan sheep as claimed in claim 1, wherein said embryo recovery from surgical embryo washing is uterine horn recovery.
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| Application Number | Priority Date | Filing Date | Title |
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| CN202210810428.7A CN115153947A (en) | 2022-07-11 | 2022-07-11 | Method for superovulation of Argwang sheep |
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| CN202210810428.7A CN115153947A (en) | 2022-07-11 | 2022-07-11 | Method for superovulation of Argwang sheep |
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Citations (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102068322A (en) * | 2011-02-22 | 2011-05-25 | 安徽农业大学 | Superovulation method for goats |
| CN104688380A (en) * | 2015-02-16 | 2015-06-10 | 中国农业大学 | Method for improving superovulating quantity and quality of sheep |
| CN107372349A (en) * | 2017-09-05 | 2017-11-24 | 灵台县现代肉牛产业（园）有限公司 | The red ox in Pingliang with and ox cross breeding method |
| CN108207797A (en) * | 2018-01-16 | 2018-06-29 | 广西大学 | A kind of efficient propagation method of goat one-child prolificacy |
| CN110151354A (en) * | 2018-01-12 | 2019-08-23 | 塔里木大学 | One kind making the oestrus increased amount of method of Sheep Ovary advantage ovarian follicle |
| WO2020061612A1 (en) * | 2018-09-24 | 2020-04-02 | Sync Innovations Pty Ltd | Method for enhancing assisted breeding techniques |
-
2022
- 2022-07-11 CN CN202210810428.7A patent/CN115153947A/en active Pending
Patent Citations (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102068322A (en) * | 2011-02-22 | 2011-05-25 | 安徽农业大学 | Superovulation method for goats |
| CN104688380A (en) * | 2015-02-16 | 2015-06-10 | 中国农业大学 | Method for improving superovulating quantity and quality of sheep |
| CN107372349A (en) * | 2017-09-05 | 2017-11-24 | 灵台县现代肉牛产业（园）有限公司 | The red ox in Pingliang with and ox cross breeding method |
| CN110151354A (en) * | 2018-01-12 | 2019-08-23 | 塔里木大学 | One kind making the oestrus increased amount of method of Sheep Ovary advantage ovarian follicle |
| CN108207797A (en) * | 2018-01-16 | 2018-06-29 | 广西大学 | A kind of efficient propagation method of goat one-child prolificacy |
| WO2020061612A1 (en) * | 2018-09-24 | 2020-04-02 | Sync Innovations Pty Ltd | Method for enhancing assisted breeding techniques |
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