Pharmaceutical composition for promoting estrus in dogs
A composition and drug technology, applied in the directions of drug combination, medical formula, vertebrate antigen components, etc., can solve the problems of low conception rate, inability to effectively induce estrus and ovulation in female dogs, and unsatisfactory effect.
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Embodiment 1
[0016] Embodiment 1: The immune effect of inhibin gene vaccine to dog
[0017] (1) Source of inhibin gene vaccine and its preparation method
[0018] The inhibin gene vaccine C501 (pVAX-S / INH-tpa) with attenuated Salmonella as the carrier was obtained from the Animal Reproduction Laboratory of the School of Animal Science and Technology of Huazhong Agricultural University. For related preparation methods, please refer to the literature: Yu Yaoyao et al., 2017). The preparation method is as follows: the correctly identified vaccine strain C501 (pVAX-S / INH-tpa) was added to 250ml LB liquid medium for expanded cultivation, and cultured with shaking at 200r / min at 37°C for 14h. Use the plasmid endotoxin removal kit to extract a large number of plasmids, measure the concentration of the plasmid, perform enzyme digestion and sequencing verification. According to needs, the obtained plasmid was diluted to different concentrations with normal saline, and stored at -20°C after aliquot...
Embodiment 2
[0037] Example 2: Inhibin Gene Vaccine Combined with PMSG on Canine Steroid Hormone Secretion
[0038] (1) The steps of preparation of vaccine, immunization and grouping of animals, and blood collection are the same as in Example 1.
[0039] (2) Determination of estrogen and progesterone:
[0040] The levels of estradiol and progesterone in canine blood were detected by double-antibody sandwich ELISA. Specific steps are as follows:
[0041] 1) Take out the ELISA plate, set a blank control well without adding any liquid; set up two wells for each standard point in turn, and add 50 μL of the corresponding standard product to each well; directly add 50 μL of the sample to be tested in each of the remaining detection wells.
[0042] 2) Add 50 μL of enzyme conjugate to each well (except for blank control wells), and then add 50 μL of antibody in the same order, mix thoroughly, attach self-adhesive to seal, and incubate at 37°C for 1 hour.
[0043] 3) Wash the plate manually and ...
Embodiment 3
[0049] Example 3: Effects of inhibin gene vaccine combined with PMSG on dog estrus and conception
[0050] Record the date of the first immunization according to the group, and closely observe each dog every day. When the vulva of the bitch is observed to be red, swollen, and dripping blood, it is judged to be in estrus. After 1 week of estrus, they were allowed to mate naturally. Record the date of estrus, breeding date, litter date and litter size. The results are shown in Table 3. According to Table 3, it can be seen that the estrus rate of bitches in T2 and T4 groups is the highest, reaching 66.7%, while the estrus rate of female dogs in the inhibin gene vaccine alone immunization group (T5) is the lowest, suggesting that inhibin gene vaccine and PMSG cooperate It can significantly increase the estrus rate of female dogs.
[0051] Table 3 Effects of different doses of inhibin gene vaccine combined with PMSG on estrus and conception in dogs
[0052]
[0053] Judging ...
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