• Journal of Embryo Transfer
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• v.27 no.1
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• pp.15-20
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• 2012

The objective of this study was to investigate the relationship between body weight, body condition score (BCS), blood urea nitrogen (BUN), glucose, cholesterol and number of transferable embryos for the purpose of improving reproductive performance in Hanwoo donors. Seventy five cows, at random stages of the estrous cycle, received a CIDR together with injection of 1mg estradiol benzoate and 50 mg progesterone, and gonadotropin treatment begann. Four days later, the animals were superovulated with a total of 28AU FSH (Antorin, 2AU = 1 ml) administered twice daily in constant doses over 4 days. On the 3rd administration of FSH, CIDR was withdrawn and 25 mg $PGF_2{\alpha}$ was administered. Cows were artificially inseminated twice after estrous detection at 12 hr intervals. The cows received $100{\mu}g$ GnRH at the time of 1st insemination. Embryos were recovered 7 days after the 1st insemination. In conclusion, cows with body weight < 400, 400~450 and > 450kg had number of transferable embryos of $4.2{\pm}1.7$, $6.1{\pm}2.7$ and $4.8{\pm}2.6$, cows with BCS <2.25, 2.25~2.75 and ${\geq}2.75$ had number of transferable embryos of $4.6{\pm}1.6$, $5.7{\pm}2.4$ and $5.1{\pm}2.7$ respectively. These data indicate that a body weight and BCS for superovulation of CIDR-treated Korean native cows does not affect the embryo yield.

• Journal of Embryo Transfer
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• v.25 no.3
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• pp.141-144
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• 2010

In vivo embryo produced from Hanwoo donor cows were collected and transferred to Hanwoo recipients. Cows, at random stages of the estrous cycle, received Progesterone Releasing Intravaginal Device (CIDR-plus, InterAg, New Zealand) together with injection of 1 mg estradiol benzoate and 50 mg progesterone, and gonadotropin treatment began 4 day later. For superovulation, a total of 28 mg FSH was intramuscularly injected twice a day in the way of decreasing doses 4 day (5, 5, 4, 4, 3, 3, 2 and 2 mg). Twenty one Hanwoo donor cows were flushed on day 7 of estrus cycle with same FSH and artificial insemination by the same technicians. Embryos were recovered 7 days after the second insemination by flushing the uterus with Embryo Collection Medium. The results obtained were as follows: The rates of transferable embryos were 50.3%, and 78 fresh embryos at morulae and blastocysts stage were transferred into Hanwoo recipients on day 7 of estrus cycle. The pregnancy rates were first embryo transfer 55.6%, 2nd 62.9% and 3rd 57.9%, respectively. In conclusion, These results suggest that CIDR-based superovulation protocol may be effectively used for production of superior Hanwoo embryos. Also, since it seems the condition of recipient cows greatly affect pregnancy rate, it is very important to evaluate recipient for effective cattle production.

• Asian-Australasian Journal of Animal Sciences
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• v.13 no.9
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• pp.1205-1209
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• 2000

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.

• Journal of Embryo Transfer
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• v.27 no.3
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• pp.127-131
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• 2012

This study was performed to investigate the effects of parity and season on the embryo production in superovulated Hanwoo cows. Superovulation was performed from 1 to 8 times by repeated superovulation treatment of Hanwoo cows (n = 22). Irrespective of estrous cycle, donor cows were received a CIDR, progesterone (50 mg) and estradiol benzoate (2.5 mg). After 4.5 days, the donor cows were superovulated with total 28AU FSH (Antorin R-10) administrated twice daily in a decreasing dose for 4 days. On $6^{th}$ and $7^{th}$ of FSH injection, 2.5 mg and 15 mg $PGF_2{\alpha}$ were injected i.m, respectively. CIDR was removed at the $7^{th}$ FSH injection. The donor cows received $200{\mu}g$GnRH at 48 hrs after $1^{st}$ $PGF_2{\alpha}$ injection. The donor cows were artificially inseminated three times after estrous detection at 12 hr intervals and embryos were recovered 7 days after estrous detection. The mean number of total ova, transferrable embryos, degenerated embryos and unfertilized oocytes were $11.6{\pm}7.9$, $5.5{\pm}4.4$, $3.0{\pm}3.3$ and $2.6{\pm}4.1$ per donor cows, respectively. A higher number of total ova were recovered in parity 3~5 ($14.3{\pm}1.3$) than 1~2 ($8.9{\pm}1.9$, P<0.05). The number of recovered normal embryos is significantly higher in parity 3~5 ($7.3{\pm}0.8$) than that of over 6 ($3.7{\pm}1.5$). Significantly higher number of total ova and normal embryos were recovered in summer ($16.4{\pm}2.3$, $8.1{\pm}1.4$) than in autumn ($10.1{\pm}1.8$, $4.5{\pm}1.1$) and winter ($6.3{\pm}1.8$, $3.3{\pm}1.1$), respectively (P<0.05). Transferable embryos were significantly higher in summer ($7.6{\pm}1.3$) than in winter ($3.0{\pm}1.0$, P< 0.05). The results were showed that parity and season affecting on the production of embryos in superovulated Hanwoo.

• Asian-Australasian Journal of Animal Sciences
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• v.16 no.7
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• pp.1071-1086
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• 2003

Reproductive biotechnologies continue to be developed for genetic improvement of both river and swamp buffalo. Although artificial insemination using frozen semen emerged some decades back, there are still considerable limitations. The major problem appears to be the lack of efficient methods for estrus detection and timely insemination. Controlled breeding experiments in the buffalo had been limited and similar to those applied in cattle. Studies on multiple ovulation and embryo transfer are essentially a replica of those in cattle, however with inherent problems such as lower number of primordial follicles on the buffalo ovary, poor fertility and seasonality of reproduction, lower population of antral follicles at all stages of the estrous cycle, poor endocrine status and a high incidence of deep atresia in ovarian follicles, the response in terms of transferable embryo recovery has remained low with 0.51 to 3.0 per donor and pregnancy rates between 15 to 30%. In vitro production of buffalo embryos is a valid alternative to recovery of embryos by superovulation. This aspect received considerable attention during the past decade, however the proportion of embryos that develops to the blastocyst stage is still around 25-30% and hence the in vitro culture procedures need substantial improvement. Embryo cryopreservation procedures for direct transfer post thaw need to be developed for bubaline embryos. Nuclear transfer and embryo cloning is a technique that has received attention in various species during recent years and can be of immense value in buffaloes as they have a low rate of embryo recoveries by both in vitro and in vivo procedures. Gender pre-selection, genome analysis, gene mapping and gene transfer are a few of the techniques that have been studied to a limited extent during recent years and are likely to be included in future studies on buffaloes. Very recently, reproductive biotechnologies have been applied to feral buffaloes as well, but the results obtained so far are modest. When fully exploited they can play an important role in the preservation of endangered species.

• Journal of Animal Science and Technology
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• v.53 no.3
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• pp.203-209
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• 2011

Lysophosphatidic acid (LPA) is a small lipid molecule that plays an important role through LPA receptors (LPARs) in reproductive processes. Our previous study has shown maximal expression of LPAR3 in the uterine endometrium on day (D) 12 of pregnancy in pigs, the period when conceptus secretes various molecules such as estrogen and interleukin-$1{\beta}$ (IL1B) and initiates implantation. We determined that endometrial expression of LPAR3 was increased by conceptus estrogen in the previous study, but the effect of IL1B on LPAR3 expression has not been determined. Thus, in this study we examined whether LPAR3 expression was also affected by IL1B. Endometrial explant cultures from D12 of the estrous cycle showed that levels of endometrial LPAR3 expression did not changed in response to IL1B. We also investigated LPAR3 expression in the uterine endometrium on D12 and D30 of pregnancy from gilts with conceptuses derived from somatic cell nuclear transfer (SCNT). The expression of LPAR3 mRNA was lower in endometria from gilts with conceptuses resulting from SCNT compared with those from gilts with embryos resulting from natural mating on D12 of pregnancy, but it was not different between them on D30 of pregnancy. Our results indicate that estrogen of conceptus origin is responsible for induction of LPAR3 expression during the peri-implantation period and appropriate LPA signaling is impaired in the uterine endometrium with SCNT-derived conceptuses during the implantation period in pigs.

• Journal of Embryo Transfer
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• v.30 no.4
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• pp.345-350
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• 2015

In the last 10 years, porcine somatic cell nuclear transfer to generate transgenic pig has been performed tremendous development with introduction and knockout of many genes. However, efficiency of porcine somatic cell nuclear transfer is still low and embryo transfer (ET) is one of important step for production efficiency. In porcine ET for production of transgenic cloned pig, we can consider many of points to increase production rates. In respect of seasonality and weather, porcine ET usually is not performed in summer and winter. Cloned transgenic embryos must be transferred into reproductive tracts of recipients where embryos are located after natural fertilization with similar estrous cycle. If cloned embryos with 2~4 cell stage are transferred, they must be transferred into oviducts in periovulatory stage. Number and deposition sites of transferred cloned embryos are important. And we must compare the methods of ET between surgical and non-surgical ones in respect of production efficiency. Sow recipients after natural estrus is most preferred recipients however its cost is must be considered. Here we will review many of current studies about porcine embryo transfer to increase production efficiency of transgenic pigs and strategies for further studies.

• Korean Journal of Veterinary Research
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• v.32 no.2
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• pp.275-280
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• 1992

To investigate the age at puberty and the seasonal breeding in Korean native goats, progesterone concentrations were measured in blood. Blood samples were collected from 8 goats at 10 day intervals from 2 months of age until the first estrus after birth, and then every 5 days for a further estrous cycle and the seasonal breeding. The mean age and weight at puberty were $195{\pm}57$ days($mean{\pm}S.D.$, range : 107~260 days) and $11.1{\pm}0.9kg$(range : 9.8~12.0kg), respectively. The mean age at first pregnancy after birth was $241{\pm}109$ days(range : 107~273 days). The estrus was observed 47.6% from October to December, and was highest in fall(38.1%) and lowest in spring and summer(14.3%). However, the estrus was observed every season. About 67% of total conception occurred form October to January. The parturition occurred 41.7% in spring, 25.0% in summer and winter, and 8.3% in fall, respectively. These results suggest that Korean native goats do not have a breeding season, but the reproductive activity is influenced by the season.

• Journal of Embryo Transfer
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• v.17 no.2
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• pp.123-128
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• 2002

The present study aimed at determining the effective dose of Folltropin, a follicle timulating hormone (FSH), on superovulation in indigenous cows of Bangladesh. Fifteen regularly cycling 5～7 years old dry cows, weighing 200～250 kg with 2.5～3.0 body condition scores (BCS) were divided into three groups (n＝5). Individual groups were superovulated with 100, 200 or 300 mg of Folltropin per animal. The superovulation treatment was initiated at Day 10 or Day 11 of the estrous cycle (Day 0＝day of estrus). Alfaprostol (6 mg) was injected to each cow 72 h after the initiation of superovulation treatment to induce eestrus. After confirming standing estrus, the cows were inseminated 2～3 times, 12 h apart, depending on the duration of estrus. At Day 6 or Day 7, individual horns of the uterus were flushed with 150～200 $m\ell$ of phosphate buffered saline supplemented with BSA (0.2％), penicillin (100 IU／$m\ell$) and streptomycin (100 $\mu\textrm{g}$／$m\ell$) using a two-way foley catheter. The embryos were concentrated, removing the excess medium through an embryo filter, and identified under a stereomicroscope. The identified embryos were collected, washed four times, evaluated and graded as excellent, good, fair or poor. The excellent, good and fair embryos were considered as transferable quality embryos. The mean (range). numbers of embryos collected vs. transferable quality embryos far 100, 200 and 300 mg of Folltropin were 4.5 (1～10) vs. 3.5 (1～8); 2.5 (1～4) vs. 1 (0～2) and 0.0 (0～0) vs. 0.0 (0～0), respectively, Folltropin at a dose of 100 or 200 mg produced suitable ovarian stimulation for superovulation in indigenous zebu cows of Bangladesh. A dose of 300 mg or more Folltropin consistently caused preovulatory corpora lutea formation in the ovaries and resulted in zero embryo recovery.

• Journal of Embryo Transfer
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• v.26 no.2
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• pp.105-109
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• 2011

Ovulation synchronization (ovsynch) has proved to increase the number of insemination in cattle by overcoming the problems of heat detection. The aim of this study was to do ovsynch in water buffaloes where heat detection is a major reproductive problem and to determine the conception rates after timed artificial insemination (TAI). Twenty cyclic buffaloes at ${\geq}$ 60 days postpartum were selected by examining 24 unobserved estrus buffaloes based on milk progesterone assay (progesterone concentration ${\geq}$ 1.0 ng/ml) from the Mymensingh district of Bangladesh. Ovsynch treatment regimen was started irrespective of the stage of estrous cycle. Gonadorelin (500 ${\mu}g$) was injected intramuscularly at Day 0 followed by Alfaprostol (8 mg) at Day 7. A second injection of Gonadorelin was given at Day 9 and TAI was done with frozen semen from Mediterranean buffalo bulls at 16~20 hours of the second Gonadorelin injection. Milk progesterone ELISA at Day 10~12 post AI confirmed ovulation in 16 out of 20 (80%) buffaloes (progesterone concentration ${\geq}$ 1.0 ng/ml). High progesterone concentration (${\geq}$ 1.0 ng/ml) at Day 10~12 and Day 22~24 of AI showed pregnancy in six out of 20 (30%) buffaloes. Pregnancy was further confirmed by ultrasonography at Day 40 in these six buffaloes. In conclusion, ovsynch followed by TAI could be applied in cyclic buffaloes for overcoming the estrus detection problems; however, more studies are needed to increase the conception rate.