阅读说明：本技术 一种山羊羔羊超数***方法 (Superovulation method for goat lambs ) 是由 吴海青 李玉荣 马跃军 于 2019-11-18 设计创作，主要内容包括：一种山羊羔羊超数排卵方法,包括：对选取的4周龄-8周龄的雌性羔羊注射总量为80IU～120IU FSH,且均分四次注射入选取的雌性羔羊体内,其中,任意相邻两次注射间隔12小时；在注射完设定总量的FSH后,间隔12小时后取卵；采用上述方法每只雌性羔羊可提供100枚以上的卵母细胞；获得的大量卵母细胞可制备大量的体外胚胎,结合胚胎移植缩短山羊育种周期,也可用于山羊卵母细胞体外培养、体外受精和胚胎体外发育等机理研究。(A method for superovulation of goat lambs, comprising: injecting 80IU-120IU FSH into the selected female lambs of 4-8 weeks old in total amount, and injecting the FSH into the selected female lambs four times in a uniform way, wherein the interval between any two adjacent injections is 12 hours; after injecting FSH with the set total amount, taking eggs after 12 hours; by adopting the method, more than 100 oocytes can be provided for each female lamb; the obtained large amount of oocytes can be used for preparing a large amount of in-vitro embryos, shortening the breeding period of goats by combining embryo transfer, and can also be used for mechanism researches such as in-vitro culture, in-vitro fertilization and embryo in-vitro development of the goat oocytes.)

1. A goat lamb superovulation method is characterized by comprising the following steps:

selecting 4-8 weeks old female lambs;

injecting 20 IU-30 IU FSH to the selected female lambs at intervals of 12 hours for 4 times;

ova were removed 12 hours apart after the fourth injection of the FSH.

2. The method of superovulation in a goat lamb of claim 1, wherein said selected female lamb is 6 weeks old and free of congenital ovarian disease.

3. The method of superovulation in a goat lamb as claimed in claim 2, wherein said FSH is injected in a total amount of 80IU-120 IU.

4. The method of superovulation in goat lambs, as claimed in any one of claims 1 to 3, wherein said FSH is injected in an amount which decreases or increases in weight depending on said selected magnetic lambs.

The technical field is as follows:

the invention relates to the technical field of animal husbandry breeding, in particular to a superovulation method for goat lambs.

Background art:

the low reproduction rate of the goats is one of the important factors restricting the development of the cashmere goat industry, and brings great difficulty to the breeding of the goats. Since the goats are single-fetus animals and ovulate 1 goat each time, the service life is 5-7 years, 4-6 lambs are produced per ewe in the whole life on average, and the breeding is relatively slow; the number of embryos obtained by superovulation technology of goat bred sheep and adult sheep is limited, and 8 effective embryos are obtained on average, so that excellent ewes cannot be fully utilized, the breeding benefit is reduced, and the breeding process is restricted; in addition, the goat oocyte is not easy to obtain from animals such as mice, rats, rabbits, pigs and the like, so that the research on the developmental mechanism of the goat embryo is slow.

The principle of lamb superovulation is that according to the special physiological characteristics that ovaries of young ewes (1-2 months old) are particularly sensitive to reproductive hormones and ovarian follicles are rarely blocked, exogenous gonadotropin is utilized to strengthen the physiological function of the ovaries, and a large number of ovarian follicles are stimulated to mature and ovulate in an estrus. Naturally, the cavitary follicles on the ovary of the fetus begin to appear at about 135 days of gestation of the ewe, the number of follicles at birth is obviously increased, then the number of follicles continues to increase until the peak is reached at 1-2 months of gestation after birth, and then the number of follicles begins to decrease with the coming of puberty. The development number of follicles on ovaries of lambs of 1-2 months after birth is not influenced by the birth season and the weight, and the phenomenon of dominant follicles on ovaries of adult lambs does not exist, so that superovulation on the lambs is a new way for efficiently producing oocytes.

A large number of scholars study the superovulation of sheep lambs to obtain a more ideal effect, Kelly et al used a similar superovulation method on 9-week-old merino lambs, and the best treatment group obtained 162.3 + -23.2 oocytes per lamb. Ledda et al (1999) hormone-treated 30-40 day old lambs: the suppository is buried in the lamb for 6 days, and 120IU GnRH and 400IUPMSG are respectively injected into the lamb once 36 hours before the suppository is removed. Eggs are collected after 24h of embolus withdrawal, and 86.2 +/-7.9 oocytes are obtained on average from each lamb. Ptak et al placed progesterone suppository 8d on 1-month-old lamb, injected 2.7mg FSH twice every morning and evening on days 5, 6 and 7, and withdrawn the suppository on day 8 to collect about 29 oocytes. But related goat lambs have fewer over-volley studies.

The superovulation of the goat lambs can fully utilize oocytes on ovaries of the ewes, and the ovum resources of high-quality dams are utilized to the maximum extent; meanwhile, the problems of insufficient embryo sources and high cost required by goat embryo transplantation are solved, and the commercial application of goat embryo production is promoted to a great extent; the lamb superovulation is combined with the in vitro fertilization technology, so that the breeding time of each generation is shortened by 2 to 3 years, if 3 generations are taken as a breeding period, the time is shortened by 6 to 9 years at least, and the genetic improvement process of goats is accelerated; in addition, the superovulation technology of the goat lambs can also be used for exploring the regulation and control mechanism of the generation of ova and the development of follicles of the goats, and other biological frontier technologies such as goat cloning, transgenic technology, establishment of embryonic stem cells and an ovum library and the like are realized.

The invention content is as follows:

in view of the above, the invention provides a goat lamb superovulation method, which aims to solve the problems in the prior art and provide technical support for shortening breeding time and enriching goat ovum resources.

In order to achieve the purpose, the invention provides the following scheme: the invention provides a goat lamb superovulation method which comprises the following steps:

selecting 4-8 weeks old female lambs;

injecting 20 IU-30 IU FSH to the selected female lambs at intervals of 12 hours for 4 times;

ova were removed 12 hours apart after the fourth injection of the FSH.

Preferably, the selected female lambs are 6 weeks of age and free of congenital ovarian disease.

Preferably, the total amount of FSH injected is 80IU-120 IU.

Preferably, the injection amount of the FSH is decreased or increased according to the body weight of the selected magnetic lamb.

The method can provide more than 100 oocytes for each female lamb; the obtained large amount of oocytes can be used for preparing a large amount of in-vitro embryos, shortening the breeding period of goats by combining embryo transfer, and can also be used for mechanism researches such as in-vitro culture, in-vitro fertilization and embryo in-vitro development of the goat oocytes.

Description of the drawings:

FIG. 1 is a schematic diagram of the method for superovulation FSH treatment of goat lambs provided by the present invention.

FIG. 2 is a diagram showing superovulation effects of goat lambs provided by the present invention.

FIG. 3 is a schematic diagram of the oocyte retrieval method provided by the present invention.

The specific implementation mode is as follows:

the technical solutions in the embodiments of the present invention will be clearly and completely described below with reference to the contents of the present invention, and it is obvious that the described embodiments are only a part of the embodiments of the present invention, and not all of the embodiments. All other embodiments, which can be derived by a person skilled in the art from the embodiments given herein without making any creative effort, shall fall within the protection scope of the present invention.

The invention aims to provide a superovulation method for goat lambs, which aims to solve the problems of long breeding period and insufficient goat egg resources of the goats.

In order to make the aforementioned objects, features and advantages of the present invention comprehensible, embodiments accompanied with figures are described in further detail below.

Referring to fig. 1 to 3 together, the present invention provides a superovulation method for a goat lamb, comprising the steps of:

1. superovulated goat lamb selection

Selecting female lambs which are strong in physique, disease-free and congenital ovarian dysfunction-free and are 4-8 weeks old, and the best lambs are about 6 weeks old; if the breeding is to be carried out, the genetic file must be complete.

2. FSH treatment method

Before FSH is injected into selected lambs, the lambs are diluted by high-fold normal saline as much as possible, so that the accurate dosage of the FSH is ensured.

Injecting the total amount of FSH to the selected female lambs at 80IU-120IU, and injecting the total amount of FSH to the selected female lambs four times, wherein the interval between any two adjacent injections is 12 h;

FIG. 1 is a schematic diagram of the goat lamb superovulation FSH treatment method, wherein the total amount of FSH injected into each lamb is 80IU-120IU, 4 times of continuous injection are carried out, 20 IU-30 IU are injected each time, the interval of each time is 12h, femoral intramuscular injection is carried out, and slow pushing is carried out.

3. Oocyte collection

48 hours after FSH is injected for the first time, the ovary volume is greatly increased due to a large number of follicles in the ovary after the lamb superovulation, and oocyte living body egg collection is preferably carried out by utilizing midline laparotomy;

with continued reference to fig. 2-3, selected female lambs are first treated for general anesthesia in an amount recommended in the specification. The egg taking process is strictly operated aseptically. Follicular fluid was aspirated using a 10ml disposable sterile syringe with # 18 needle until the follicular fluid was aspirated. Passing the oocyte-containing oocyte, follicular fluid and oocyte preservation fluid through a sterile filter screen of 100 μm, washing with the oocyte preservation fluid for 3-5 times, and picking up the oocyte under a microscope for subsequent work.

According to statistics, the average number of follicles in ovaries at both sides of 100 goat lambs treated by the method is 103 +/-8.3, and P is less than 0.01. Since the number of collected oocytes is related to the proficiency of the operation steps of egg aspiration and the like of an operator, the number of the obtained oocytes is increased along with the improvement of the proficiency of the egg aspiration operator.

The principle and the implementation mode of the invention are explained by applying a specific example, and the description of the embodiment is only used for helping to understand the method and the core idea of the invention; meanwhile, for a person skilled in the art, according to the idea of the present invention, the specific embodiments and the application range may be changed. In view of the above, the present disclosure should not be construed as limiting the invention.