在來山羊의 發情週期와 發情持續期間을 조사하고자 정관 절제 시술한 시정모 산양을 이용하여 90두의 암산양으로부터 3년간에 걸쳐 총 148회의 방정주기를 관찰하였다. 총 481회의 발정주기의 평균기간은 18.1${\pm}$0.5이었고, 11일 이하의 短發情週期가 143회(30%), 12일에서 16일의 中發情週期가 15회(3%), 17일에서 24일의 正常發情週期가 268회(56%) 및 25일 이상의 長發情週期가 55회(11%)로 나타났다. 短發情週期의 발생율은 자연발정후(27%)보다 유기발정후(15.2${\pm}$0.8)일에 유의적(P<0.01)으로 짧았다. 短發情週期에 있어서는 143회중 6일의 기간을 가지는 빈도가 92회로 가장 많았다. 발정지속 시간의 평균은 34.0${\pm}$0.5시간이었는데, 발정지속시간에 있어서는 자연발정과 유기발정간에 유의적인 차이가 없었다. 이상의 결과로 在來山羊의 發情週期는 自然發情後에 있어서 보다 誘起發情間에 차이가 없음을 알 수 있다.
To study the conditions to enhance success of embryo transfer in the dog, 20 mixed-breed bitches were used for the experiment along with 4 male dogs for mating. The bitches were paired according to synchronism of natural estrus, or the counterpart as donor or recipient was treated with gonadotropin as FSH (follicular stimulating hormone) or PMSG (pregnant mare serum gonadotropin) for induction of estrus to be synchronized with estrus of the other bitch in natural estrus. Embryo recovery was performed in two ways for comparison, either by flushing each uterine horn after ovariohysterectomy or by flushing each horn in the state of non-ovariohysterectomy. In addition, the result of pregnancy according to the embryo stage and the repeatability of the experimental animals as donor or recipient were also investigated. FSH or PMSG was administered to the bitches which had passed over 4 months from last estrus, resulting in estrus-positive in 3 dogs of 6 FSH-treated dogs (50.0%), and in 5 dogs of 9 PMSG-treated dogs (55.6%), determined by proestrus signs and vaginal smear test. Estrus-positive bitches induced with gonadotropin were used as donor or recipient resulting in one embryo-recovered bitch as donor and one offspring-delivered bitch as recipient in 5 PMSG-treated dogs, whereas no result was obtained from 3 FSH-treated dogs. The rate of embryo recovery to be compared with number of corpus luteum was 68.2% in ovariohysterectomized dogs and 55.2% in non-ovariohysterectomized dogs, respectively. The number of dogs from which embryo was collected were 4 dogs of 6 ovariohysterectomized dogs (66.7%) and 6 dogs of 7 non-ovariohysterectomized dogs (85.7%), respectively. The result of parturition was obtained from one dog of 5 estrus-induced recipients, whereas no result was obtained from 3 natural-estrus recipients. The only dog which delivered a male puppy had been transferred 3 morulae and 2 blastocysts. Of 6 repeat-used bitches in canine embryo transfer, 3 dogs showed repeatability either as donor or recipient. These results indicated that inducing estrus of a dog with gonadotropin is feasible in canine embryo transfer to be synchronized with that of a natural-estrus dog, that embryo recovery is also possible in non-hysterectomized dogs, that the estrus-induced dog is also usable as recipient to result in parturition, and that repeat-use of a bitch as donor or(and) recipient is possible in canine embryo transfer.
Estrus was induced and the therapeutic effect was examined with 25mg of PGF2$\alpha$ intramuscular injection and 3mg of PGF2$\alpha$ intraovarian injection to Holstein and Simmental cows which were raised in the large size stockfarms and the dairy farms, and were diagnosed to anestrus. The results obtained were summarized as follows: 1. The estrus inducing effect observed in the cows treated by PGF2$\alpha$ was 83.3% with 25/30 heads in the intramuscular injection group, and 86.7% with 26/30 heads in the intraovarian injection group. 2. Average duration from PGF2$\alpha$ administration to the onest of estrus was 2.7 day in the intramuscular injection group, and 2.6 days in the intraovarian injection group. 3. In status of estrus in the cows treated by PGF2$\alpha$, vigorous estrus was 12.0%, normal estrus 60.0% and silent estrus was 28.0% in the intramuscular injection group, and vigorous estrus was 15.4%, normal estrus 61.5%, and silent estrus 23.1% in the intraovarian injection group. 4. Conception rate in the cows induced to estrus was 64.0% in the intramuscular injection group and 65.4% in the intraovarian injection group.
Growth hormone (GH) is obligatory for growth and development. But, there is controversy on the GH effect about reproductive processes of sexual differentiation, pubertal maturation, gonadal steroidogenesis, gametogenesis and ovulation. This study was conducted to investigate the effect of GH on estrus, ovulation and embryo implantation. The results obtained were as follows. GH stimulated to increase estrus rate (p<0.05), pregnancy rate (p<0.05), and total fetus number in mice treated for superovulation. Also, the correlation between GH and steroids, E2 and P4, at peri-estrus stage/ peri-ovulation stage/ peri-implantation stage of the superovulation-induced mice was examined. Consequently, GH co-injected with PMSG especially increased P4 level (p<0.05) at peri-estrus stage of superovulationinduced mice. In conclusion, GH co-treatment in superovulation system boosted the rate of estrus, pregnancy and total fetus by increasing progesterone level at peri-estrus stage of superovulation-induced mice.
Considerable attention has been focused on the cryopreservation of semen and estrus induction in dog, as consequence of poor productivity caused by long anestrus period, in order to enhance the productivity of youngs and to preserve the breeds. The objectives of this study were to evaluate semen quality after cryopreservation and to evaluate the Pregnancy rate after insemination (AI). Fifty infertilie dogs (age 2∼3 years) were selected for the study and divided into three different estrus induction treatment groups. Group 1: dogs (n=15) were given clomifene (0.1 mg/kg) orally for five days at 12 hr intervals. Group 2: dogs (n=15) were given bromocriptine (50 $\mu\textrm{g}$/kg) orally for five days at 12 hr intervals, followed by single injection intravenously of 500 IU GnRH (Group 3, n=20) when pro-estrus occurred. The rates of pregnancy in estrus inducted dogs mated naturally compared to those inseminated artificially with ejaculated fresh semen and frozen-thawed semen. Estrus detection was performed using the method of vaginal smear and confirmed by the plasma progesterone assay. The ejaculated semen to freeze was exposed to a mixture of Tris extender with cryoprotectant (Trisma, 81 mM: TES, 209 mM: citric acid, 6 mM; glucose, 5 mM; glycerol, 8%) and cryopreserved gradually by slow-cooling at 17 cm above the surface of liquid nitrogen (LN$_2$) for 23 min. The motility of frozen-thawed spermatozoa was assessed by phase-contrast microscopy. To assess their viability and acrosome content, spermatozoa were stained with a vital stain and Fluorescence conjugated lectin Pisum Savitum Agglutinin (FITC/PAS), respectively. Pregnancy was confirmed by ultrasonograpy on day 25, 35 and 55 post insemination. The use of fresh semen, the pregnancy rates were observed 66.6, 66.6, 75.0 and 83.3% in natural estrus, clomifene induced, bromocriptine induced and a combination of GnRH and bromocriptine, respectively. The use of frozen-thawed semen, the pregnancy rates were observed 66.6, 33.3, 50.0 and 60.0% in natural estrus, clomifene induced, bromocriptine induced and a combination of GnRH and bromocriptine, respectively. No difference was observed in the number of offspring produced among natural estrus and treated groups inseminated with fresh or frozen-thawed semen. In conclusion, the pregnancy rate of dogs treated with a combination of GnRH and bromocriptine was more effective than use of clomifene or bromocriptine only. In addition, frozen-thawed semen can be used successfully far artificial insemination in dog.
It has been well known that estrogens stimulate the uterine contractility and progestins inhibit it. Then, one may expect that the uterine contractility and sensitivities to oxytocin (OT) and prostaglandin $F_{2{\alpha}}\;(PGF_{2{\alpha}})$ would be different among the estrus cycle. These hypotheses were tested using the mature female rat. Spontaneous isometric contractions of isolated uterine strips $(1{\times}0.3\;cm)$ from cyclic rats in various stages of the estrus cycle, bilateral ovarectomized rats and hypophysectomized rats were recorded in absence or presence with $estradiol-17{\beta}\;(E_2)$, progesterone $(P_4)$, OT and $PGF_{2{\alpha}}$. The results were summarized as follows: 1) The spontaneous uterine contractile force was the highest in the estrus rat and the lowest in the ovarectomized or the hypophysectomized rat. In the proestrus rat, the contractile frequency was the lowest (2.7 beats/10 min) and the contractile duration was the longest (70 sec). In the other groups, there were no any differencies in frequency (9 beats/10 min) and in duration (30 sec). 2) OT and $PGF_{2{\alpha}}$ stimulated the uterine contractility in all groups tested except in the hypophysectomized rat in which OT failed to stimulate the uterine contraction. $PGF_{2{\alpha}}$ was more effective in stimulating the uterine contraction than OT in all groups tested except in the estrus rat. OT-induced contraction was the highest in the estrus rat and $PGF_{2{\alpha}}-induced$ contraction was the lowest in the hypophysectomized rat. 3) Uterine contractilities were not changed by the in vitro treatments of $E_2$ or $P_4$ under the influence of endogenous steroids, however, $E_2$ and $P_4$ stimulated the uterine contraction in the ovarectomized rat in which endogenous steroids were almost abolished. 4) Increased uterine contraction by the treatment of OT was suppressed by in vitro $E_2$ or $P_4$ in the estrus rat, while it was potentiated by the $P_4$ in the proestrus rat. In other groups, exogenous $E_2$ or $P_4$ did not affect the OT-induced uterine contraction. 5) $PGF_{2{\alpha}}-induced$ uterine contraction was suppressed in the ovarectomized rat by $E_2$ and $P_4$, in the diestrus and proestrus rats by $P_4$ and in the hypophysectomized rat by $E_2$. In other groups, exogenous $E_2$ or $P_4$ was ineffective in altering the $PGF_{2{\alpha}}-induced$ uterine contraction. According to the above results, it may conclude that the mechanisms of the different uterine contractility and the different uterine sensitivity to OT or $PGF_{2{\alpha}}$ according to the estrus cycle are not explicable with only the serum concentrations of steroids, OT and $PGF_{2{\alpha}}$ but also other unknown factors.
The present study was undertaken to investigate the effect of stimulation of follicular development with eCG on the peripheral levels of inhibin and FSH in Murrah buffaloes. Estrus was synchronized in five normally cycling females by insertion of Crestar (Intervet, Boxmeer, Holland) implants for nine days. Estradiol valerate was administered i.m. on the day of implant insertion. On the 10th day of the induced estrous cycle a single dose of 3000 IU eCG (Folligon, Intervet, Boxmeer, Holland) was given, followed by treatment with 25 mg of $PGF_2$ alpha (Lutalyse, Upjohn, Belgium) 48 h later. Blood samples were obtained during the induced estrus, on cycle day 10 (luteal phase), at the superovulatory estrus (43 h after PGF) and during the periovulatory period (64 h after PGF). Ultrasonography was done daily to monitor follicular development. Plasma concentrations of inhibin and FSH were determined by specific radioimmunoassays. Differences between $mean{\pm}SEM$ values of different phases of the cycle were compared by ANOVA. The mean number of small (2-5 mm), medium (6-9 mm) and large (>10 mm) follicles observed two days after eCG treatment and on the day of superovulatory estrus was $2.8{\pm}0.31$, $5.2{\pm}0.30$ and $1.4{\pm}0.09$ and $1.9{\pm}0.21$, $2.8{\pm}0.40$ and $5.0{\pm}0.83$, respectively. The mean number of ovulations was $3.6{\pm}0.37$ and the mean number of unovulated follicles was $6.1{\pm}0.47$. Most of the follicles >10 mm in diameter had ovulated (72%). The mean ${\pm}SEM $ of plasma inhibin concentration $(2584.15{\pm}17.92pg/ml)$ during the superovulatory estrus was significantly higher $(p{\leq}0.05)$ than during the induced estrus $(749.87{\pm}17.29pg/ml)$, the luteal phase $(1099.54{\pm}24.98pg/ml)$ and periovulatory period $(1682.71{\pm}29.88pg/ml)$, respectively. $Mean{\pm}SEM$ plasma FSH concentration during the induced estrus $(10.35{\pm}0.41ng/ml)$ was not different from that during the superovulatory estrus $(8.52{\pm}0.39ng/ml)$, but was significantly higher $(p{\leq}0.05)$ than during the luteal phase $(2.81{\pm}0.42ng/ml)$ and periovulatory period $(5.7{\pm}0.28ng/ml)$. These data indicate that treatment with eCG in buffaloes for inducing superovulation results in a significant elevation in plasma inhibin levels and a decrease in plasma FSH levels during the superovulatory estrus. Thus, we suggest that the elevated plasma inhibin coming from fully developed follicles continued for a long time which results in inhibition of FSH leading to poor ovulation in the remaining follicles, which may be the cause of suboptimal superovulatory response.
This study was performed to investigate the patterns of progesterone secretion after induction of estrus in premature, metestrous and anestrous bitches. A total of 22 bitches were used. Of them 18 bitches were treated with hormone to induced estrus and 4 bitches were untreated and served as controls. Estrus was induced with $PGF_{2{\alpha}}$, estrone, estradiol-$17{\beta}$, PMSG and HCG(Treatment A), and with PMSG and HCG(Treatment B). Blood samples were collected via the cephalic vein at 2 to 5 days interval. Blood samples were centrifuged (1,200g, 10min.) within 30 minutes after collection and plasma was stored at $-20^{\circ}C$ until analyzed for the progesterone concentrations. Plamsa progesterone concentrations were measured by radioimmunoassay. The results of estrous induction were determined by estrous signs, ovarian response, egg recovery and progesterone patterns. The results obtained were as follows; 1. All bitches in treatment A showed estrous signs, however the ovarian response and egg recovery were not detectable and the levels of progesterone were nearly same as before. 2. In the treatment B, premature and metestrous bitches showed only estrous signs, however 5 of 7 anestrous bitches (71.4%) showed estrous signs, ovarian response and changes of progesterone levels. In conclusion, clinical estrous behavior can be induced during any phase of the estrous cycle, but ovulation should be induced only if induction occur approximately 4 months or more after the previous estrus.
The aim of the present recent study was to compare the protein patterns in the vaginal mucus of Hanwoo cattles during spontaneous and CIDR induced-estrus. Ten cattles, who had been observed in estrus, received no treatment and served as the group of cattles with normal spontaneous estrus. Thirteen cattles in the CIDR received an CIDR insert on day 14 were removed and cattles were injected GnRH on day 15. Vaginal mucus samples were collected from all cattles at the same time the single AI in cattles with spontaneous estrus and the AI in cattles with induced estrus. Spontaneous and CIDR-induced estrus vaginal mucus samples were analyzed on two different array surfaces: cation-exchange (CM10), anion-exchange (Q10). In addition, using the NaCl solution by which the proteins combined after washing are 0.5, 1 and 2 M, it was fractionated and a protein was collected successively. The results are summarized as follows: 1) Ionic surfaces chemistries (Q10 and CM10) gave the best results in terms of detectable protein peaks, with more than 100 protein peaks in the two fractions and under each condition. 2) Protein mass spectrometer using 11 different proteins in protein identification of 7 were able to determine the protein. List of identified proteins as follows; Ribosome-binding protein 1, GRIP 1-associated protein 1, Katanin p60 ATPase-containing subunit A-like 1, Protein FAM44A, DUF729 domain-containing protein 1, Prolactin precursor, Dihydrofolate erductase. Conclusively, on the basis of this study, protein expression in the vaginal mucus could be used as an indicator for time of estrus manifestation in order to increase conception rates by applying AI at an optional time.
Estrus was induced in 13 anestrus Korea Jin-do bitches by intramuscular injection of pregnant mare serum gonadotropin (PMSG) in a dose of 500 lU once daily for ten consecutive days, followed by an additional single intraveneous injection of 1,000 lU of human chorionic gonadotropin (hCG) on the tenth day. Day-changes of vaginal epithelial cells during the hormone treatment were investigated in each experimental bitches and compared with the those of spontaneous estrus bitches. The first days of vulval bleeding and male acceptance after PMSG treatment were on Day 6.0$\pm$ 1.5 (mean$\pm$ SD) and Day 9.0$\pm$ 1.9, respectively. And in all of 13 bitches, vulval swelling and perineal reflex were shown. The mean durations of proestrus and estrus were 2.9$\pm$ 1.4 (mean$\pm$ SD, range ; 1-6) and 11.5: 1.7 (range ; 8-14) days, respectively, that is, duration of proestrus was significantly shorter than that of the spontneous estrous bitches but duration of estrus was longer than that of the spontaneous estrous bitches. Characteristic features of vaginal cytology during the estrous cycle were the high proportions of large intermediate cell, superficial cell, anuclear cell and erythrocyte in proestrus, superficial cell and anuclear cell in estrus and parabasal cell, small intermediate, large intermediate cell, and leukocyte in diestrus, respectively. The comification index (Cl) was significantly high proportion in proestrus and estrus, when Day 0 was timed from the first day of male acceptance, the Cl was first increased above 80% on Day 0 and maintained above 80% until Day 0 to Day 5 during 6 days and showed a peak on Day 2. Also it was maintained above 90% until Day 2 to Day 3 during 2 days. These results indicated that all 13 ekperimental bitches showed positive estrus detection by the estrus behavior and vaginal smear test after treated with PMSG and hCC. It suggested that vaginal cytology was used to estimate the optimal mating and ovulation time, in consideration of the day when the Cl was maintained above 80% in estrus-induced Korea Jin-do bitches.
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