CN111134084A - Method for optimizing timing insemination effect of replacement gilts - Google Patents
Method for optimizing timing insemination effect of replacement gilts Download PDFInfo
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- CN111134084A CN111134084A CN202010010220.8A CN202010010220A CN111134084A CN 111134084 A CN111134084 A CN 111134084A CN 202010010220 A CN202010010220 A CN 202010010220A CN 111134084 A CN111134084 A CN 111134084A
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- 229960000971 altrenogest Drugs 0.000 claims description 8
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- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01K—ANIMAL HUSBANDRY; AVICULTURE; APICULTURE; PISCICULTURE; FISHING; REARING OR BREEDING ANIMALS, NOT OTHERWISE PROVIDED FOR; NEW BREEDS OF ANIMALS
- A01K67/00—Rearing or breeding animals, not otherwise provided for; New or modified breeds of animals
- A01K67/02—Breeding vertebrates
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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
- A61D19/02—Instruments or methods for reproduction or fertilisation for artificial insemination
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61D—VETERINARY INSTRUMENTS, IMPLEMENTS, TOOLS, OR METHODS
- A61D7/00—Devices or methods for introducing solid, liquid, or gaseous remedies or other materials into or onto the bodies of animals
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Abstract
The invention discloses a method for optimizing the timing insemination effect of replacement gilts, which comprises the steps of continuously feeding replacement gilts of the right age to allergol 18d with the dosage of 20 mg/head/day, injecting 10ml AIS/head into the muscle of the replacement gilts after the allergol is stopped for 42h, injecting 200 mu g GnRH/head into the muscle of the replacement gilts after 80h, performing first artificial insemination after 24h, and performing second artificial insemination after 16 h. The AIS of the invention replaces PMSG in the prior timed insemination procedure, can obviously increase the ovulation number of replacement gilts, reduce the ovarian cyst rate and improve the hormone secretion condition of the replacement gilts, realizes the optimization of the prior timed insemination technology and further improves the reproductive performance of the replacement gilts implementing the FTAI procedure.
Description
Technical Field
The invention relates to the technical field of livestock breeding, in particular to a method for optimizing the timing insemination effect of replacement gilts.
Background
The replacement gilt is used as a storage resource of a pig farm virtuous cycle, and the quality and reproductive capacity of the replacement gilt directly influence the production level and economic benefit of the pig farm. In a large-scale pig farm, replacement gilts are supplemented every year by 20-30% of basic sows. However, the rate of oestrus failure of replacement gilts after sexual maturity or body maturity is 15% to 20%. Therefore, the method improves the oestrus and ovulation of replacement gilts, enables the replacement gilts to successfully enter a production sow group, and furthest excavates the potential of the replacement gilts, and becomes an urgent problem to be solved.
At present, a technology for artificially controlling and adjusting the process of the oestrus cycle of a sow by using a hormone preparation so as to concentrate oestrus, ovulation and hybridization within a preset time is a Fixed Time Artificial Insemination (FTAI) technology, and the technology can effectively solve the problem of low oestrus rate of replacement sows. The sow breeding management technology taking the timing insemination technology as the core gradually becomes the technical basis for realizing batch production management in large-scale pig farms. However, in the existing technical mode, a timing insemination procedure has certain uncertainty in optimizing breeding indexes such as conception and farrowing performance of replacement gilts, and the influence of exogenous hormone treatment on replacement gilts, ovulation, embryonic development and the like still needs to be further researched.
In the current timed insemination procedure, Pregnant Mare Serum Gonadotropin (PMSG) is often adopted as an inducer for superovulation, but because PMSG has a long half-life period, fertilization and embryonic development after ovulation can be interfered, and repeated stimulation of PMSG has risks of causing follicular cyst (or luteinization) and drug resistance of livestock, so that the reproductive performance of sows is negatively influenced. Meanwhile, the duration of the PMSG for promoting the follicular development to mature ovulation and the ovulation effect have strong individual difference, and are also one of the factors for limiting the wide application of the timing insemination technology. Therefore, finding a PMSG alternative that reduces or eliminates these adverse effects may help to achieve an optimization of the effectiveness of existing timed insemination techniques.
Disclosure of Invention
The invention aims to provide a method for optimizing the timing insemination effect of replacement gilts, so as to overcome the defects in the prior art.
The invention adopts the following technical scheme:
a method for optimizing the timing insemination effect of replacement gilts comprises the steps of continuously feeding replacement gilts of the right age to allylpregnensin 18d with the dose of 20 mg/head/day, injecting 10ml AIS/head into muscles of the replacement gilts 42h after feeding of the allylpregnensin, injecting 200 mu g GnRH/head into muscles after 80h, performing first artificial insemination 24h later, and performing second artificial insemination 16h later.
Further, the replacement gilts with the suitable age are replacement gilts with the age of 210 +/-3 days.
The invention has the beneficial effects that:
1. the AIS replaces PMSG in the existing timing insemination procedure, follicle development is more effectively induced, ovulation rate and embryo development potential of replacement gilts are improved, and optimization of the existing timing insemination technology is achieved. According to the method, the AIS is used for replacing PMSG in the existing timing insemination procedure to induce follicular development, and the ovarian state, ovulation rate, embryo development potential and reproductive hormone secretion condition of the replacement gilt after oestrus and mating of the replacement gilt are contrastively analyzed, so that the result shows that the AIS replaces PMSG in the existing timing insemination procedure to remarkably increase the ovulation number of the replacement gilt, reduce the ovarian cyst rate and improve the hormone secretion condition of the replacement gilt, the optimization of the existing timing insemination technology is realized, and the reproductive performance of the replacement gilt implementing the FTAI procedure is improved.
2. After the AIS replaces PMSG in the existing timing insemination procedure, the replacement gilt with ovarian cyst is reduced in number and increased in ovulation number, wherein the ovulation number of the replacement gilt is improved by 58.8%, and the ovarian cyst rate is reduced by 20.8%; and under the condition that no obvious difference is found in the total recovered embryo number, the 6-8cell number is obviously increased, and the embryo development is promoted to a certain extent.
3.E2The level may reflect follicular development. The AIS replaces PMSG in the prior timed insemination procedureReplacement of E in sow serum2Level is markedly elevated, E2The level is improved by 1.36 times in the process of inducing estrus, which shows that the invention has more effective promotion effect on the follicular development of replacement gilts, and the result is consistent with the result that the red body number of the replacement gilts in the improved group is obviously higher than that of the replacement gilts in the control group.
4.P4Plays an important role in establishing and maintaining the pregnancy process. The AIS of the invention replaces P in the serum of the backup sow after PMSG in the prior timed insemination procedure4Level is markedly elevated, P4The content is improved by 1.35 times after 48 hours of mating, and is more beneficial to embryo implantation, development and pregnancy maintenance.
Drawings
FIG. 1 is a flow chart of hormone treatment.
FIG. 2 shows the ovarian status comparison of each group.
FIG. 3 shows the developmental status of recovered embryos.
FIG. 4 is a graph of embryonic DAPI staining.
Detailed Description
The invention is explained in more detail below with reference to exemplary embodiments and the accompanying drawings. The following examples are provided only for illustrating the present invention and are not intended to limit the scope of the present invention.
1 laboratory animal
Randomly selecting 17 Jinhua two WuxDuroc hybrid replacement gilts with the age of 210 (+ -3) days and the weight of about 110kg as experimental animals, and performing induced feeding for 4-5 days by using fruit juice before the experiment is performed so that the experimental animals can adapt to a drug feeder.
2 test grouping and procedure
(1) Grouping ① control group by using the existing FTAI program, and ② improved group by replacing 1000IUPMSG in the existing FTAI program with 10ml of Anti-inhibin serum (AIS).
(2) The process comprises the following steps: altrenogest was fed daily at 3 pm, Day1 of altrenogest was recorded as Day1, followed by 18 consecutive days of altrenogest at 20 mg/head/Day. According to the difference of the injection medicines to the replacement gilts after stopping feeding the altrenogest for 42h, the 17 replacement gilts are randomly divided into: 1) a control group (n-9) administered by intramuscular injection of 1000IUPMSG (available from Sanbo Biotech, Inc., Ningbo) 42h after withdrawal of altrenogest; 2) trial modified group (n-8) 10ml AIS/head (purchased from RIKEN) was injected intramuscularly 42h after withdrawal of altrenogest. After 80h, 200. mu.g GnRH were injected intramuscularly to each test sow. And carrying out first artificial insemination 24h later, and carrying out second artificial insemination 16h later. Finally, the test sows are sacrificed 48h after the second artificial insemination, and venous blood is collected for detecting the contents of estradiol and progesterone in serum; collecting ovaries, oviducts and uteruses, and counting the red body number, ovarian cyst number, ovarian diameter, oviduct length, uterine horn length, recovered embryo number and embryo number of different development stages of each sow; and continuously culturing the collected fertilized eggs, and observing the number of blastula developed by each sow and the blastula rate. The specific experimental procedure is shown in FIG. 1.
3 collecting and detecting samples
(1) Estradiol (E) in serum2) And progesterone (P)4) And (3) content detection: collecting blood (5 ml/head) from anterior vena cava of each group of replacement sows at 4 time points of the final Day of allylogest feeding (Day18), the current Day of intramuscular PMSG/AIS (Day 20), the current Day of intramuscular GnRH (Day23), the current Day of slaughtering and the like; the blood sample is kept at 4 ℃ overnight, centrifuged at 4000r/min at 4 ℃ for 15min, the serum is separated and stored at-20 ℃. The serum sample (1 ml/head) is sent to Hangzhou gold region medical inspection institute, Inc., and all the groups of reproductive hormone E are detected and analyzed2And P4The variation of (2).
(2) And (3) analyzing the ovary state: the size of the ovary on both sides is measured by observing and counting each group of red bodies (the number of ovulation points) and calculating according to the length of the maximum contact surface multiplied by the width.
(3) And (3) embryo recovery: washing oviduct and uterus of each group of replacement gilts in a tube respectively by PBS containing 1% FBS at constant temperature of 37 ℃, recovering early embryos, observing the development states (single cell, two cells, four cells and the like) of the embryos by microscopic examination, and counting the number of the embryos in each development period; the length of the oviduct and uterine horn after embryo retrieval was measured.
(4) Embryo in vitro culture: collecting embryos of each group of the same individual by using a mouth suction tube, uniformly transferring the embryos into an embryo preservation micro-tube, and carrying the embryos back to a laboratory from a slaughterhouse for embryo culture at a constant temperature of 37 ℃.
(5) And (3) counting embryos: when in vitro culture is carried out for 3d, early-stage development blastula in each group are randomly selected for embryo counting, and the main operation steps are as follows: ensuring that the number of the effective blastula in each group is more than 10; and continuously culturing the rest embryos, observing the development state of each group of blastocysts, and counting the development rate of the blastocysts.
4 results of the test
(1) The effect on the ovarian status of replacement gilts after AIS treatment is shown in fig. 2, with significantly more red bodies on the ovaries of the improved group (fig. 2A) than the control group (fig. 2B); the ovaries of both the improved group and the control group had large follicles with a diameter of more than 6mm, but the number of red bodies in the ovaries containing both large follicles and red bodies was larger in the improved group than in the control group. The number of the red bodies, the size of the ovary, the length of the oviduct, the length of the uterine horn and the number of ovarian cysts of each group are counted, and the results are shown in table 1: the red body number of the improved group is 1.59 times of that of the control group, and is obviously higher than that of the control group (p < 0.05). No significant difference was observed in ovarian size and fallopian tube length between the two groups (p > 0.05); the uterine horn length of the improved group was significantly lower than that of the control group (p < 0.05); in addition, 1 ovarian cyst was observed in the modified group sows, while 3 ovarian cysts were observed in the control group sows.
TABLE 1 comparison of Red body number, ovarian size, fallopian tube and uterine horn length
(2) The effect of AIS treatment on ovulation and fertilization in replacement gilts is shown in table 2 and fig. 3, and the results show that there was no statistically significant difference between the average number of embryos retrieved from the sows in the improved group and the average number of embryos retrieved from the control group (p >0.05), but the average number of embryos retrieved from each sow in the improved group was 4-5 more than that in the control group. Meanwhile, both the improved group and the control group had embryos/oocytes in a 1-cell state, and it was confirmed that these 1-cells were unfertilized oocytes through in vitro culture, wherein the proportion of unfertilized ova in the control group was 27.5% higher than the proportion of unfertilized ova in the improved group by 9.17%. In addition, the number of embryos developed to 6-8cell stage in the improved group was also significantly higher than that in the control group (p < 0.01).
TABLE 2 recovery of embryos from groups
(3) The influence of the AIS treatment on the development of replacement gilt embryos is shown in figure 4 and table 3, and the results show that DAPI staining is carried out when the embryos recovered from each group are cultured in vitro to the 3 rd day (figure 4), and then the development conditions of the blastula are counted, so that the blastula development rates of the fertilized embryos of the improved group and the control group are found to be not significantly different (p is greater than 0.05) and are 78.04 +/-4.24% and 77.86 +/-6.39% respectively; the cell numbers of early blastocysts were 45.0. + -. 2.80 and 43.25. + -. 1.55, respectively, and there was no significant difference (p >0.05) (Table 3).
TABLE 3 embryonic development
(4) The influence of the AIS treatment on the reproduction hormone level of replacement gilts is shown in tables 4 and 5, and the detection and analysis of the hormone level in blood of each group at four time points show that the estrogen level of the improved group is remarkably improved by 1.36 times (p is less than 0.001) compared with the control group in the process of inducing estrus; the progesterone content in the serum of the sow prepared by the improved group 48 hours after the hybridization is also obviously improved by 1.35 times (p is less than 0.001).
TABLE 4 serum E of the later sows2Content (wt.)
TABLE 5 serum P of the later sows4Content (wt.)
In the existing timed insemination procedure, PMSG is often used as an inducer for superovulation, but researches show that ovarian cyst can be caused when PMSG is used for superovulation stimulation, normal ovulation time is interfered, embryo development is inhibited, and adverse effects such as drug resistance of livestock are caused. The AIS is used for replacing PMSG in the existing timing insemination procedure, and the result shows that the average red body number of the improved group is obviously higher than that of the control group and has the advantage of being averagely higher than 10, so that the AIS has obvious advantage in actual pig raising production and also reflects that the ovulation effect of replacement gilts can be obviously improved through the AIS. Meanwhile, compared with a control group, the number of embryos in 6-8cell stages in the improved group is obviously increased, and the phenomenon that replacement sows of the improved group generate ovarian cysts is also obviously improved, so that the method effectively promotes the development of the embryos and improves the condition of higher ovarian cyst rate in the conventional timed insemination procedure. In addition, no obvious difference is found in the diameter of the ovary and the length of the fallopian tube between the two groups, which indicates that the AIS replaces PMSG in the existing timed insemination procedure to a certain extent and has no influence on the development of the ovary and the fallopian tube. Compared with the invention, the replacement gilt of the control group has relatively longer uterine horn length; however, while PMSG can increase uterine horn length, studies have shown that PMSG can delay embryonic development, implantation, and lead to prolonged gestational time. Therefore, the present invention has significant advantages over the prior timed insemination procedure.
Research shows that most follicles are finally locked in the follicle development process, and only partial dominant follicles can effectively ovulate; and in the process E2The level may reflect follicular development. The present invention shows that, in comparison to the current timed insemination procedure, the present invention, in use, provides a replacement for E in the blood of sows2The content is obviously higher than that of the control group, which shows that the invention has more effective promotion effect on the follicular development of replacement gilts, and the result is also corresponding to the result that the red body number of the improved group is obviously higher than that of the control group. At the same time, P4Plays an important role in establishing and maintaining the pregnancy process, relatively high concentration of P4Is more beneficial to embryo implantation, fetal development and the like, and compared with the prior insemination procedure, the invention has the advantages that the insemination procedure is carried out to a certain extentCan more obviously improve P in the blood of replacement gilts4Content, which also indicates that the replacement gilt using the present invention has a more favorable gestational environment.
Compared with the existing timed insemination procedure, the method can effectively improve the ovulation number of replacement gilts, reduce the occurrence probability of ovarian cysts and provide a superior hormone internal environment for embryo implantation, development and pregnancy maintenance.
Claims (2)
1. A method for optimizing the timing insemination effect of replacement gilts is characterized in that replacement gilts of the right age are continuously fed with the altrenogest 18d with the dose of 20 mg/head/day, 10ml of AIS/head is injected into the muscle of the replacement gilts after the feeding of the altrenogest is stopped for 42 hours, 200 mu g of GnRH/head is injected into the muscle of the replacement gilts after 80 hours, first artificial insemination is performed after 24 hours, and second artificial insemination is performed after 16 hours.
2. The method of optimizing gilt replacement gilts for timed insemination as recited in claim 1 wherein said gilt replacement gilts are replacement gilts 210 ± 3 days old.
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Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN111713452A (en) * | 2020-07-06 | 2020-09-29 | 浙江省农业科学院 | Method for improving timing insemination efficiency of replacement gilts |
| CN112089503A (en) * | 2020-09-29 | 2020-12-18 | 湖北省农业科学院畜牧兽医研究所 | Method for in vitro cloning of embryo transplantation surrogate pregnant sow |
| WO2023109803A1 (en) | 2021-12-15 | 2023-06-22 | 宁波三生生物科技股份有限公司 | Antibody against inhibin and use thereof |
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| WO2023109803A1 (en) | 2021-12-15 | 2023-06-22 | 宁波三生生物科技股份有限公司 | Antibody against inhibin and use thereof |
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