CN106474133A - A kind of preparation improving milch cow conception rate and its application - Google Patents
A kind of preparation improving milch cow conception rate and its application Download PDFInfo
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- CN106474133A CN106474133A CN201610837114.0A CN201610837114A CN106474133A CN 106474133 A CN106474133 A CN 106474133A CN 201610837114 A CN201610837114 A CN 201610837114A CN 106474133 A CN106474133 A CN 106474133A
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- melatonin
- prostaglandin
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- milch cow
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K31/00—Medicinal preparations containing organic active ingredients
- A61K31/557—Eicosanoids, e.g. leukotrienes or prostaglandins
- A61K31/5575—Eicosanoids, e.g. leukotrienes or prostaglandins having a cyclopentane, e.g. prostaglandin E2, prostaglandin F2-alpha
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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
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- 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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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K31/00—Medicinal preparations containing organic active ingredients
- A61K31/33—Heterocyclic compounds
- A61K31/395—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins
- A61K31/40—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having five-membered rings with one nitrogen as the only ring hetero atom, e.g. sulpiride, succinimide, tolmetin, buflomedil
- A61K31/403—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having five-membered rings with one nitrogen as the only ring hetero atom, e.g. sulpiride, succinimide, tolmetin, buflomedil condensed with carbocyclic rings, e.g. carbazole
- A61K31/404—Indoles, e.g. pindolol
- A61K31/4045—Indole-alkylamines; Amides thereof, e.g. serotonin, melatonin
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- Life Sciences & Earth Sciences (AREA)
- Animal Behavior & Ethology (AREA)
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- Reproductive Health (AREA)
- Medicines Containing Material From Animals Or Micro-Organisms (AREA)
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Abstract
The invention provides a kind of improve the preparation of milch cow conception rate and its application, the effective ingredient of described preparation contains melatonin and prostaglandin.Its using method is to add semen deposition after preparation mixing after semen thawing.The present invention is tested by milch cow and finds, described preparation can improve the conception rate after milk frozen cattle semens semen deposition, can have good market prospect and social effect in livestock culturing field wide popularization and application.
Description
Technical field
The present invention relates to the improving technology field of domestic animal artificial insemination is and in particular to a kind of preparation of raising milch cow conception rate
And preparation method and application.
Background technology
Technology of artificial insemination is very good, the highly developed know-how of an effect in livestock reproduction, in propulsion kind
Improvement, improve and improve the output of live-stock product and quality aspect meaning is very great.The technology of artificial insemination of milch cow is from 20th century 80
Age starts to widely use in China, and subsequent frozen semen gradually instead of fresh essence insemination.Recently as scale, intensive
Popularization, the milk yield of milch cow is increasing year by year, but the reproductive efficiency of milch cow is but declining.After how improving milch cow semen deposition
Conception rate and then the reproductive efficiency of raising milch cow, become the focus of breeding scholar's research.
Prostaglandin (prostaglandin) finds in the seminal fluid be present in people earliest, essentially from seminal vesicle.Hereafter find
From animal body Various Tissues, such as testis, uterus, Placenta Hominiss and ovary etc. are all extractable.Its mainly have dissolving corpus luteum function with
And promoting smooth muscle contraction, this transports in reproductive tract to the wriggling in uterus, afterbirth discharge in puerperal, sperm when heat, ovulation, childbirth
OK, all have a certain impact.At present, prostate F2α(PGF2α) and the like be efficiently used for the estrus synchronization of milch cow, hold
The treatment of the ovarian disease such as corpus luteum, corpus lutein cyst long.(2015, the Comparison of various semen such as Abdussamad
extenders and addition of prostaglandin F2αOn pregnancy rate in cows) research show,
Add 1.5mg/ml PGF in frozen semen2α, conception rate can reach 56%.A kind of patent of invention (side of raising cow conception rate
Method, application number, 201510265568.0) 1 to 2 hour before artificial insemination, suck 0.2mg prostaglandin 2ml, note with deferent duct
It is mapped to cervix uteri crotch, the conception rate of milch cow can be improved.
Melatonin (melatonin) is a kind of neuro-endocrinology hormone mainly secreted by pinuies, and chemical constitution is N-
Acetyl -5-MT 5-methoxytryptamine (N-acetyl-5-methoxytryptamine), it is in the side such as biological clock, reproduction, immunity, antioxidation
Face plays an important role.Meanwhile, also there are some researches show, melatonin can directly participate in growth and the ovulation of follicle, improve cattle ovum female
Cell maturation rate and the developmental rate of embryo.Patent of invention (a kind of preparation of raising milch cow conception rate and preparation method thereof with should
With, application number, 201510085043.9) 10-3～10-1The melatonin of mol/L and concentration are 10-6～10-4The Vitamin E of mol/L
Milch cow conception rate can be effectively improved.
Improve milch cow conception rate method and formulation also have a lot, but the lifting amplitude of milch cow conception rate still have very big
Space.And, a lot of technical operations are complicated, need through professional training, the popularization that have impact on technology to a certain extent should
With.
Content of the invention
It is an object of the invention to provide a kind of preparation improving milch cow conception rate and its application.
Present invention research has been surprisingly found that, under the collective effect with prostaglandin for the melatonin, the conception rate of embryo is entered
The raising of one step, based on this, technical scheme is as follows：
First, a kind of preparation improving domestic animal conception rate of the present invention, described preparation is mixed solution, and its effective ingredient is to take off
Melanocyte and prostaglandin.
Preferably, described melatonin and the weight ratio of prostaglandin are：1:6250-625000, it is further preferred that described melatonin
Weight ratio with prostaglandin is：1:62500.
In described formulation soln, the concentration of described melatonin is preferably 4.8-480ng/ml, more preferably 48ng/
ml.The concentration of described prostaglandin is preferably 2-4mg/ml, more preferably 3mg/ml.
It will be appreciated by those skilled in the art that the concentration of above-mentioned preparation can be concentrated or dilute according to the needs of commodity
Release, when using, preparation can use after accordingly being diluted.In addition, the solvent of formulation soln can be this area can accept
Any can be used for diluting the solvent of sperm, for example conventional spermatozoa diluent.System for convenience of statement melatonin and prostaglandin
It is referred to as MP preparation (Melatonin-Prostaglandin agent) under agent.
Secondly, the present invention provides a kind of method improving domestic animal conception rate, and it is by the melatonin of effective dose and prostaglandin
Collective effect treats pregnant domestic animal.Described effective dose be directed to different domestic animal dosage can difference, for example, can for its dosage of sheep
In 50-200 μ L, can be 100-500 μ L for horse dosage, can be 100-500 μ L for cattle dosage.Side for effect
Formula can be directly injected into, for example by described preparation be injected into intrauterine, Uterus wall or other close to uterus position.When
So, for different plant species and different methods, dosage can be adjusted according to the conception rate of test.A kind of enforcement of the present invention
Mode is the melatonin measuring effect and prostaglandin is mixed after mixed liquor with seminal fluid and carries out artificial insemination.Preferably, described
In mixed liquor, the concentration of melatonin is 2.4-240ng/ml, more preferably 24ng/ml；Preferably, the concentration of described prostaglandin
For 1-2mg/ml, more preferably 1.5mg/ml.
Finally, the present invention provides the application on improving domestic animal conception rate of melatonin and prostaglandin, and described MP
Application on improving domestic animal conception rate for the preparation.
The MP preparation of the raising milch cow conception rate that the present invention provides, is that melatonin is molten in certain proportion with prostaglandin
Solution is in semen diluent, and mixes, with the frozen semen after thawing, the conception rate carrying out improving milch cow after semen deposition.The party
Method is simple to operation, by the raising of conception rate, and then improves the reproductive efficiency of milch cow, reduces cost, increased economy
Benefit, therefore has a good application prospect.
Specific embodiment：
Following examples are used for further illustrating the present invention, but should not be construed as limitation of the present invention.Without departing substantially from this
On the premise of spirit and essence, made for the present invention modify or replace, belong to the scope of the present invention.Ground promise prostatitis
Parathyrine is all purchased from Minitueb purchased from Pfizer, semen diluent, and melatonin is purchased from Sigma.
Embodiment 1 improves MP preparation of milch cow conception rate and preparation method thereof
Raw material：Dinoprostin, melatonin, semen diluent
Preparation method：
1) weigh 8.16mg melatonin, be dissolved in the dehydrated alcohol of 1mL, dilute 1000 times with PBS, become final concentration
8.16ng/ μ L, standby.
2) in 0.5mL EP pipe, it is separately added into the Dinoprostin 102 μ L that concentration is 5mg/ml, semen diluent 67
μ L, melatonin 1 μ L, 170 μ L MP preparation altogether, mix.
In MP preparation, the concentration of melatonin is 48ng/ml, and the concentration of Dinoprostin is 3mg/ml.
All operations all complete in darkroom.
Embodiment 2 improves MP preparation of milch cow conception rate and preparation method thereof
Raw material：Dinoprostin, melatonin, semen diluent
Preparation method：
1) weigh 81.6mg melatonin, be dissolved in the dehydrated alcohol of 1mL, dilute 1000 times with PBS, become final concentration
81.6ng/ μ L, standby.
2) in 0.5mL EP pipe, it is separately added into the Dinoprostin 136 μ L that concentration is 5mg/ml, semen diluent 33
μ L, melatonin 1 μ L, 170 μ L MP preparation altogether, mix.
In MP preparation, the concentration of melatonin is 480ng/ml, and the concentration of Dinoprostin is 4mg/ml.
All operations all complete in darkroom.
Embodiment 3 improves MP preparation of milch cow conception rate and preparation method thereof
Raw material：Dinoprostin, melatonin, semen diluent
Preparation method：
1) weigh 8.16mg melatonin, be dissolved in the dehydrated alcohol of 1mL, dilute 10000 times with PBS, become final concentration
0.816ng/ μ L, standby.
2) in 0.5mL EP pipe, it is separately added into the Dinoprostin 68 μ L that concentration is 5mg/ml, semen diluent 101
μ L, melatonin 1 μ L, 170 μ L MP preparation altogether, mix.
In MP preparation, the concentration of melatonin is 4.8ng/ml, and the concentration of Dinoprostin is 2mg/ml.
All operations all complete in darkroom.
Embodiment 4 semen thawing, is mixed with MP preparation
1) take out milch cow 0.25mL straw frozen semen (freezing essence purchased from the property control of Beijing Milk Cow Center) from liquid nitrogen, be placed in 38 DEG C
Thaw 30 seconds in water-bath；
2) cut off tubule sealing, frozen semen added in the EP pipe added with 170 μ L MP preparations of 38 DEG C of preheatings, mixes,
Make mixed solution.In mixed liquor, the final concentration of 24ng/ml of melatonin, the concentration of Dinoprostin is 1.5mg/ml.Separately
If matched group, melatonin experimental group and Dinoprostin experimental group, matched group be semen diluent, melatonin experimental group and ground
Promise prostaglandin experimental group only contains melatonin and Dinoprostin respectively.Each group operation is same.Specifically it is grouped as follows：
The content of melatonin group and Dinoprostin in each group seminal fluid mixed liquor of table 1
| Group | Melatonin (ng/ml) | Prostaglandin (mg/ml) | Cumulative volume (μ L) |
| Matched group | 0 | 0 | 340 |
| Melatonin group | 24 | 0 | 340 |
| Dinoprostin group | 0 | 1.5 | 340 |
| Melatonin group+Dinoprostin group | 24 | 1.5 | 340 |
3) mixed liquor is all added in the semen thin tube of 0.5mL, loads spermatic gun, prepare semen deposition；
4) selection of semen deposition object
Preferably 15-20 monthly age, body weight reach more than 350 kilograms, estrus cycle normal yearling heifer, because of growing cattle reproduction
Function is vigorous, uterus environment is good, is conducive to being impregnated.
5) determination of Inseminated time
It is strict controlled in preovulatory 3-5 hour semen deposition, after that is, milch cow accepts mounting, 12-14 hour carries out examination per rectum,
It is optimal Inseminated time that follicle has one to touch when breaking.And combine visual inspection, milch cow performance is quiet, and swelling of external genitals starts to disappear,
Vaginal orifice outflow is translucent to lead thread mucus.
6) semen deposition method
Strict sterilization cow pudendum.Using the semen deposition of body of uterus deep, insemination site requires 2-5 at the pull of ovulation Aconitum carmichaeli Debx. palace
Centimeters, so ensure have enough sperms to reach and ovum insemination position.And accomplish " gently insert, push away slowly, delay " during semen deposition, with
Exempt from the uterus of cow to be caused damage.
7) diagnosis of early gestation
Breed latter 28 days, gather blood 5ml, separate serum, using Ai Deshi visual early pregnancy detection kit, check milch cow
Fertilization situation, and statistical result.
Conception rate after each group seminal fluid mixed liquor semen deposition of table 2
| Process | Process milk ox head number | Pregnant head number | Conception rate (%) |
| Matched group | 87 | 42 | 48.28±2.07a |
| Melatonin group | 65 | 34 | 52.31±1.12b |
| Dinoprostin group | 59 | 32 | 54.24±1.35b |
| Melatonin group+Dinoprostin group | 82 | 56 | 68.29±2.60c |
Statistical result showed：Said preparation is remarkably improved the conception rate after milch cow breeding.
Conclusion：Understand that melatonin and Dinoprostin are used in combination and can improve being subject to after milch cow semen deposition by result above
Tire rate.Therefore, this composite reagent can be applicable to the external semen deposition of milch cow.
Claims (10)
1., it is characterised in that described preparation is mixed solution, its effective ingredient is black for taking off for a kind of preparation improving domestic animal conception rate
Element and prostaglandin.
2. preparation according to claim 1 is it is characterised in that described melatonin with the weight ratio of prostaglandin is：1:
6250-625000.
3. preparation according to claim 1 is it is characterised in that described melatonin with the weight ratio of prostaglandin is：1:
62500.
4. preparation according to claim 1 is it is characterised in that the concentration of described melatonin is 4.8-480ng/ml, preferably
For 48ng/ml.
5. preparation according to claim 1 is it is characterised in that the concentration of described prostaglandin is 2-4mg/ml, preferably
3mg/ml.
6. a kind of method improving domestic animal conception rate, the melatonin of effective dose and prostaglandin collective effect are treated pregnant domestic animal by it.
7. method according to claim 6 it is characterised in that mix the melatonin of effective dose and prostaglandin with seminal fluid
Carry out artificial fertilization after making mixed liquor.
8. method according to claim 7 it is characterised in that in described mixed liquor melatonin concentration be 2.4-240ng/
Ml, preferably 24ng/ml, the concentration of described prostaglandin is 1-2mg/ml, preferably 1.5mg/ml.
9. the melatonin and prostaglandin application on improving domestic animal conception rate, the preparation described in any one of claim 1-5 exists
Improve the application on domestic animal conception rate.Described domestic animal is milch cow.
10. the prostaglandin described in claim 1-9 is Dinoprostin.
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Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN109287621A (en) * | 2018-11-16 | 2019-02-01 | 翼城县富华养殖有限公司 | A kind of extender of freezing semen improving milk cow conception rate |
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Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN109287621A (en) * | 2018-11-16 | 2019-02-01 | 翼城县富华养殖有限公司 | A kind of extender of freezing semen improving milk cow conception rate |
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