CN110495967A - A kind of method of same period ovulation timing semen deposition - Google Patents
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- 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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Abstract
The invention discloses a kind of methods of ovulation timing semen deposition of same period, it includes: the 1) 4d before ox oestrus any one day, are 10 ~ 20mg/50kg Niu Tichong epiphysin to ox subcutaneous doses；2) 0d is to ox intramuscular injection 80 ~ 200 μ g GnRH, 7d to ox 0.1 ~ 0.5mg of intramuscular injection PG；3) after crossing 50 ~ 60h, to 80 ~ 200 μ g GnRH of intramuscular injection of ox, subcutaneously injection dosage is the epiphysin of 10 ~ 20mg/50kg Niu Tichong, and artificial insemination；Pregnancy rate reaches 75% after being inseminated with this method, and MDA concentration reduces in the cow blood of measurement, SOD, GSH-PXIt is increased with CAT concentration；The invention has the advantages that: 1) this method save detection of oestrus semen deposition before；2) superfecundation, gestation can unify the cultivation of lactation period management, the scientific feeding management for doing calf, growing cattle well on a large scale of concentrating one's energy.
Description
Technical field
The invention belongs to technical field of animal husbandry, and in particular to a kind of method of same period ovulation timing semen deposition.
Background technique
China's beef cattle breeding mode, which is based primarily upon, puts in a suitable place to breed and drylot feeding.Based on stocking model is put in a suitable place to breed with mountain area mountainous region, the cultivation
For mode due to factors such as early springs, post-partum estrus breeding rate is lower, and puts in a suitable place to breed on Niu Shan, after estrus of cow, hands over naturally
Match, seriously limits the popularization of beef cattle technology of artificial insemination and the raising of reproductive efficiency.Drylot feeding is main in beef cattle breeding supports
Grow mode.Cow is since long-term stable breeding leads to malnutrition, microelement deficiencies, amount of exercise deficiency etc., so that it is numerous to influence cow
Grow ability.Mainly causes estrus of cow symptom unobvious or only ovulate and out of heat, the follicular development of persistence corpus luteum inhibition and hair
Situations such as feelings.
Currently, beef cattle industries continue propulsive factor transition, casual household accelerates to exit, and large-scale cultivation is gradually formed, with from numerous
The mode of autotrophy has become the important symbol of beef cattle industries structure optimization adjustment.And low reproduction rate is that restriction beef cattle industries are fast
The bottleneck of speed development.Estrus of cow symptom is unobvious, and duration of estrus is long, especially recessive heat, false heat, continuous estrus,
The presence of the abnormal phenomenon such as out of heat causes cow entirety calving rate low, and feeding cost is high, and economic benefit of aquaculture is low.For solution
The certainly drawback in beef cattle production, using same period ovulation timing insemination technique (FTAI) using Exogenous Reproductive to estrus of cow
Ovulation carries out same period regulation, can control using Exogenous Reproductive ox heat and row in desired extent according to certain programs
Ovum, and the Fixed Time Interval that can be identified in no oestrus carries out artificial insemination, and it helps to improve conception rate
With breeding rate, shorten the calving interval, reduces feeding cost investment, improve the production efficiency of cows, promote the fast of beef cattle industries
Speed development.The application of same period ovulation timing insemination technique helps to improve beef cattle reproductive efficiency.Domestic related ox FTAI technology
It studies relatively fewer and is mainly applied on milk cow, rarely applied on beef cattle.There are research and production practices to be proved FTAI skill
Beef cattle reproductive efficiency can be improved in art.The present invention improves the ovulation timing semen deposition of the beef cattle same period by optimizing to Cosynch program
Technical level, promote beef cattle industries fast-developing.
Although same period ovulation-timing insemination technique is able to achieve high efficiency quick breeding, but its efficiency is still to be improved, at present the skill
The pregnancy rate that art obtains after applying in beef cattle is 40%~60% or so.Patent CN105613423A " domestic animal same period ovulation-fixed
When semen deposition method " in disclose domestic animal same period ovulation-timing semen deposition method: 1) in any one day of domestic animal oestrous cycle muscle
Inject GnRH or its analog；2) PG or its analog are injected after 7-8 days；3) GnRH or its analog are injected after 2-3 days again；
It is specifically a kind of same period ovulation-timing semen deposition method of milk cow, is 45.45% using the conception rate that this method reaches,
The technical effect reached does not improve a lot.Therefore need to provide a kind of method that conception rate is high, to guarantee beef cattle breeding effect
Rate.
Summary of the invention
Object of the present invention is to not high to solve cattle-raising production cycle length, the conception rate of existing same period ovulation-timing semen deposition
The problem of, and a kind of method of the same period that conception rate is high ovulation timing semen deposition is provided.
A kind of method of same period ovulation timing semen deposition, it includes:
1) 0d, -4d taking off for 10 ~ 20mg/50kg Niu Tichong to ox subcutaneous doses were used as by the ox oestrous cycle any one day
Melanocyte；
2) 0d is to ox intramuscular injection 80 ~ 200 μ g GnRH, 7d to ox 0.25 ~ 0.5mg of intramuscular injection PG；
3) after 7d crosses 50 ~ 60h, to 80 ~ 200 μ g GnRH of intramuscular injection of ox, subcutaneously injection dosage is 10 ~ 20mg/50kg
The epiphysin of ox weight, and artificial insemination；
The injection dosage of the epiphysin is 10 ~ 15mg/50kg Niu Tichong；
The injection volume of the GnRH is 80 ~ 120 μ g；
The injection volume of PG described in step 2 is 0.25mg；
After 7d described in step 3) crosses 56h, to 80 ~ 200 μ g GnRH of intramuscular injection of ox.
The present invention provides a kind of methods of ovulation timing semen deposition of same period, it includes: 1) in ox oestrous cycle any one day
4d(-4d before (0d)), the epiphysin for being 10 ~ 20mg/50kg Niu Tichong to ox subcutaneous doses；2) 0d is infused to ox muscle
80 ~ 200 μ g GnRH, 7d are penetrated to ox 0.25 ~ 0.5mg of intramuscular injection PG；3) after crossing 50 ~ 60h, the intramuscular injection 80 ~ 200 to ox
μ g GnRH, subcutaneously injection dosage is the epiphysin of 10 ~ 20mg/50kg Niu Tichong, and artificial insemination；After being inseminated with this method
Pregnancy rate reaches 75%, and MDA concentration reduces in the cow blood of measurement, SOD, GSH-PXIt is increased with CAT concentration, CAT activity change
It is not significant；The invention has the advantages that: 1) this method save detection of oestrus semen deposition before；2) superfecundation, gestation can be unified to feed
The management of newborn phase is cultivated, and can concentrate one's energy the scientific feeding management for doing calf, growing cattle well on a large scale.
Detailed description of the invention
Fig. 1 estrus synchronization Fixed Time Insemination program schematic diagram；
Influence of Fig. 2 synchronization of Estrus to black ox cow pregnancy rate；
Fig. 3 processing is to P in the 10th d serum after black ox cow artificial insemination4The influence of concentration；
Fig. 4 processing is to P in the 45th d serum after black ox cow artificial insemination4The influence of concentration；
Fig. 5 influence of the front and back injection MT to MDA concentration in heifer blood for the first time；
Fig. 6 influence of the front and back injection MT to SOD concentration in heifer blood for the first time；
Fig. 7 influence of the front and back injection MT to GSH-Px concentration in heifer blood for the first time；
Fig. 8 influence of the front and back injection MT to CAT concentration in heifer blood for the first time；
Influence before and after second of injection MT of Fig. 9 to MDA concentration in heifer blood；
Influence before and after second of injection MT of Figure 10 to SOD concentration in heifer blood；
Influence before and after second of injection MT of Figure 11 to GSH-Px concentration in heifer blood；
Influence before and after second of injection MT of Figure 12 to CAT concentration in heifer blood.
Specific embodiment
A kind of method of ovulation timing semen deposition of the same period of embodiment 1
A kind of method of same period ovulation timing semen deposition, it includes:
1) the young Simmental for taking health, the growth of livestock moderate (Body Condition Score is between 2.5-4.0), through ultrasound diagnosis when beginning
Ovary basic condition is divided into luteal phase (corpus luteum diameter > 0.3 cm), follicular phase (follicular diameter > 0.3 cm), resting stage (no Huang
Body and ovarian follicle), reproductive organs is normal, without reproductive diseases；
2) oestrous cycle any one day is determined as 0d, from 0d several 4d(-4d forward) subcutaneous injection epiphysin, injection dosage is
12mg/50kg Niu Tichong；
3) 0d is to ox intramuscular injection GnRH(100 μ g), from 0d several 7d(+7d backward) to ox intramuscular injection PG(0.25mg),
To intramuscular injection GnRH(100 μ g of ox) and subcutaneous injection epiphysin (12mg/50kg Niu Tichong) after 56h, and manually award
Essence.
The ovulation timing semen deposition experiment of the 2 beef cattle same period of embodiment
1, experimental animal
The age is chosen in the young west gate tower of the 18 more than monthly age 86 health, the growth of livestock moderate (Body Condition Score is between 2.5-4.0)
That ox, all test ox, through ultrasound diagnosis ovary basic condition, are divided into luteal phase (corpus luteum diameter > 0.3 in on-test
Cm), follicular phase (follicular diameter > 0.3 cm), resting stage (no corpus luteum and ovarian follicle), reproductive organs is normal, without reproductive diseases.
2, it is grouped
86 test ox picked out are randomly divided into 4 groups: control group (CTR, n=25), takes off at an epiphysin group (MT, n=20)
Melanocyte+HCG group (MT+HCG, n=21), twice epiphysin group (2MT, n=20) (Fig. 1)；
1) control group: 0d first time intramuscular injection GnRH(100 μ g), 7d intramuscular injection PG(0.25mg), second after 56h
Intramuscular injection GnRH(100 μ g) and artificial insemination (AI)；
2) an epiphysin group (MT group): epiphysin is subcutaneously injected in -4d, and injection dosage is 12 mg/50kg Niu Tichong；0d muscle
Inject GnRH(100 μ g), 7d intramuscular injection PG(0.25mg), intramuscular injection GnRH(100 μ g after 56h) and artificial insemination
(AI)；
3) epiphysin+HCG group: epiphysin is subcutaneously injected in -4d, and injection dosage is 12 mg/50kg Niu Tichong；0d intramuscular injection
GnRH(100 μ g), 7d intramuscular injection PG(0.25 mg), intramuscular injection GnRH(100 μ g after 56 h) and artificial insemination
(AI), intramuscular injection HCG(1000 unit after artificial insemination 5d)；
4) epiphysin group twice: epiphysin is subcutaneously injected in -4 d, and injection dosage is 12 mg/50kg Niu Tichong；0 d intramuscular injection
GnRH(100 μ g), 7d intramuscular injection PG(0.25mg), intramuscular injection GnRH(100 μ g after 56h) and subcutaneous injection epiphysin
(12mg/50kg Niu Tichong) and artificial insemination (AI)；
Then plus physiological saline the described epiphysin preparation is first to be dissolved with dehydrated alcohol, and, ethyl alcohol and physiological saline ratio are 7 ratios
3, it is configured to the solution of 60mg/ml；
The 0d refers to ox oestrous cycle any one day, and -4d is 4 days before first time (0 day) intramuscular injection GnRH.
3, raising and sample collection
The test ox and other non-test oxen picked out individually divide circle to raise, and every circle raises 5 to 6 oxen.Using fully mixed diet
TMR daily ration is prepared in feeding every time when feeding, concentrate feeding twice respectively at two time points of morning 09:00 and 16:00 in afternoon,
Free water；
10 oxen of every group of random selection are injected before test process, after injection epiphysin (- 4d) for the first time for 24 hours, for the second time respectively
Before epiphysin (7d+56h), after second injection epiphysin (7d+56h) for 24 hours, six times of 10d and 45d after artificial insemination
Point tail vein acquires 10mL blood sample；Blood sample collection in the non-anticoagulant heparin tube of vacuum, place 4 DEG C of refrigerators for 24 hours after its solidification after, make
It is centrifuged 10min with blood centrifugal machine 3000rpm, upper serum is transferred in 1.5mL sterile centrifugation tube with liquid-transfering gun, freezing is protected
It is stored in -20 DEG C of refrigerators, it is spare.
4, artificial insemination
1) preparation before inseminating: 75% alcohol wipe of metal inseminating syringe is sterilized, and every ox of inseminating syringe housing uses disposable
Housing；Insemination person will wear test clothes, and nail cannot be too long, and a Zhi Shouxu of insertion cow rectum wears disposable rectum inspection Check
Gloves (outside suds wetting gloves)；The cow that will receive insemination is fixed, and tail is fixed on side on Shelf for keeing, with new
Geramine solution cleans cow external genital；
2) semen thawing inspection: straw frozen semen is placed in 40 DEG C of thermostatted waters and stands about 20 seconds；It is living with microscope inspection Check sperm
Power, objective table temperature should be maintained at 40 DEG C, and sperm motility can be used 0.3 or more；
3) semen deposition: being inserted into disposable plastic tubule for inseminating syringe push rod, has on one end insertion inseminating syringe push rod of tampon, will be another
End cuts off sealing；The hand of insemination personnel is inserted into the rectum of cow to be inseminated to catch cow uteri neck, and another hand will fill
There is the inseminating syringe of sperm to be inserted into cow uteri neck mouth to be inseminated by cow vagina, is then injected into sperm.
5, cyesiognosis
Rectum gestation detection time 60d after testing cows' breeding is carried out；Be pregnant 60 d cow cornua uteri slightly about not
2 times of Pregnant cows cornua uteri, the cornua uteri of Pregnant cows is almost liquid filled, and has fluctuation, and empty angle only has a semiliquid,
Cornua uteri inter-drain flattens smooth, but can differentiate two corners turnoff, can touch whole uterus；Sometimes the tire of the size such as home mouse can be touched
Youngster, touching fetal membrane can feel there are 3 blood vessels；Cyesiognosis work is assisted to complete by cattle farm technical staff.
As a result as shown in table 1 and Fig. 2, twice epiphysin group heifer pregnancy rate be significantly higher than control group and epiphysin+
HCG group (CTR vs 2MT, P=0.036；MT+HCG vs 2MT, P=0.017), compared with an epiphysin group, difference is not significant
(P=0.053).
6, progesterone and oxidative stress Indexs measure
Test progesterone (P4), malonaldehyde (MDA), superoxide dismutase (SOD), glutathione peroxidase in cow serum
(GSH-PX) it is all made of enzyme-linked immunosorbent assay (ELISA) with catalase (CAT) level and is detected, according to kit
Kit after 4 DEG C of refrigerators taking-ups, is placed room temperature by operation instructions, balances 30 min, and test serum takes from -20 DEG C of refrigerators
Out, it balances to room temperature.20x concentrated cleaning solution in kit is diluted to 1x for future use with distilled water.
1) progesterone level changes
After artificial insemination in 10d and 45d each group cow serum progesterone level variation as shown in Figures 3 and 4, progesterone level twice
Variation tendency is identical；Epiphysin+HCG group progesterone level is significantly higher than control group and an epiphysin group (P < 0.05), and twice
Epiphysin group is compared, and progesterone level difference is not significant (P > 0.05)；
2) MDA and antioxidase change before and after injection epiphysin for the first time
MDA result of variations is as shown in figure 5, before the injection of first time epiphysin, the not significant (P of MDA concentration difference in each group cow blood
>0.05) after, injecting 24 h of epiphysin, the MDA concentration of MT group, MT+HCG group and 2MT group be substantially lower than control group (P<
0.05)；CAT, SOD and GSH-Px concentration variation tendency are as shown in Fig. 6 ~ 8 in each group cow blood, and each group is female before epiphysin is injected
Three kinds of antioxidase content differences are not significant (P > 0.05) in cow's serum, after injecting 24 h of epiphysin, MT group, MT+HCG group and
The SOD and GSH-P of 2MT groupXConcentration is all significantly higher than control group (P < 0.05).It is injected after injection epiphysin with first time for the first time
Preceding comparison each group CAT content is on the rise, but injects preceding not significant (P > 0.05) with each group contrast difference after injection；
3) MDA and antioxidase change before and after second of injection epiphysin
MDA result of variations is as shown in figure 9, MDA concentration and antioxidase change in cow blood before and after second of injection epiphysin
Trend is similar to variation tendency before and after first time injection epiphysin.Before second of epiphysin injection, each group MDA concentration difference is not shown
It writes (P > 0.05), it is on the rise compared with before injection for the first time, but difference is not significant (P > 0.05), second of injection epiphysin
After 24 h, 2MT group MDA concentration is substantially less than MT group, MT+HCG group and CTR group (P < 0.05)；As shown in Figure 10 ~ 12, second
Three kinds of antioxidase content differences are not significant in each group cow serum before secondary epiphysin is injected, under having compared with before first time injection
Drop trend, but difference is not significant (P > 0.05).After injecting 24 h of epiphysin, SOD and GSH-P in cow blood in 2MT groupXIt is dense
Degree is significantly higher than MT group, MT+HCG group and CTR group (P < 0.05).Comparison is each before injecting after second of injection epiphysin with second
CAT concentration is on the rise in group cow blood, but injects preceding not significant (P > 0.05) with each group contrast difference after injection.
Claims (6)
1. a kind of method of same period ovulation timing semen deposition, it includes:
1) ox oestrous cycle any one day is set as 0d, -4d to ox subcutaneous doses be 10 ~ 20mg/50kg Niu Tichong
Epiphysin；
2) 0d is to 80 ~ 200 μ of ox intramuscular injection g/ GnRH, 7d to 0.25 ~ 0.5mg of ox intramuscular injection/head PG；
3) after crossing 50 ~ 60h, to intramuscular injection g/ GnRH of 80 ~ 200 μ of ox, subcutaneously injection dosage is 10 ~ 20mg/50kg
The epiphysin of ox weight, and artificial insemination.
2. a kind of method of same period ovulation timing semen deposition according to claim 1, it is characterised in that: the epiphysin
Injection dosage is 10 ~ 15mg/50kg Niu Tichong.
3. a kind of method of same period ovulation timing semen deposition according to claim 2, it is characterised in that: the note of the GnRH
The amount of penetrating is 80 ~ 120 μ g.
4. a kind of method of same period ovulation timing semen deposition according to claim 3, it is characterised in that: PG described in step 2
Injection volume be 0.25mg.
5. a kind of method of same period ovulation timing semen deposition according to claim 4, it is characterised in that: mistake described in step 3)
After 56h, to the intramuscular injection GnRH of ox.
6. a kind of according to claim 1, method of same period ovulation timing semen deposition described in 2,3,4 or 5, it is characterised in that: described
Epiphysin first dissolved with dehydrated alcohol, then plus physiological saline, ethyl alcohol and physiological saline volume ratio are 7 to 3.
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| CN112370421A (en) * | 2020-10-16 | 2021-02-19 | 中国农业大学 | Composition for improving milk quality and preparation method and application thereof |
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Application publication date: 20191126 |