• Journal of Embryo Transfer
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• v.29 no.3
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• pp.283-287
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• 2014

Sexed semen is commonly used for the production of calves of the desired gender. Gender selection is important in animal production industries. For example, female cattle are required for the dairy industry while males are preferred in the beef cattle industry. The present study was to assess the in vivo embryo production efficiency using the semen separated according to sex during superovulation in Hanwoo. Seventy Hanwoo donor cows were flushed on day 7 of estrus cycle with same FSH and artificial insemination by the same technicians. Embryos were recovered on 7 days after the third insemination by flushing the uterus with embryo collection medium. KPN semen straws used artificial insemination contained 20 million sperm (total number 60 million per donor). Sex-sorted semen straws contained 4 million sperm (total number 12 million per donor). The results obtained were as follows: No differences were observed in the efficiency of superovulation rates on KPN semen 87%, and sexed semen 100%, respectively. The mean numbers of total embryos are each $12.58{\pm}8.31$ and $13.25{\pm}7.86$. The mean numbers of transferable embryos, sexed semen were significantly lower than KPN semen ($3.75{\pm}1.98$ vs. $8.23{\pm}6.07$, P<0.05). The rates of unfertilized embryos from superovulation using sexed semen were significantly higher than KPN semen (50% vs. 15%, P<0.05). The rate of degenerated 2-cell embryos from sexed and KPN semen was 60.87% and 11.11%, respectively (p<0.05). In conclusion, these results indicate that superovulation using sexed semen was useful, but efficient embryo production was important to reducing the damage caused by the Flowcytometer-based sperm sorting procedure.

• 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.

• Journal of Embryo Transfer
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• v.32 no.3
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• pp.171-176
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• 2017

This study was conducted to investigate the breeding status of farms to improve the production efficiency of Hanwoo calf. The study was conducted on 45 farms divided into two groups. This study was conducted to investigate the breeding size and breeding area of Hanwoo cows. The average age at first delivery of Hanwoo was 28.7 months. The number of artificial insemination per pregnancy was $1.45{\pm}0.32$, and the number of artificial insemination days after birth was 119.8 days. Conception rates were $75.2{\pm}16.93%$ for small farms and $70.6{\pm}17.46%$ for medium sized farms and $71.4{\pm}11.03%$ for large farms. When we looked at farming methods, 'the farmers using estrus observation aids' had 10.42% higher calf production rate than the 'unused farmers'. The farms vaccinated with IBR and BVDV for breeding cattle showed a 4.41% decrease in abortion, stillbirth and mortality. According to farming conditions, conception rate and delivery rate improved by 3.47% and 18.29%, respectively, when grazing and exercising were performed. Observation, immunization and grazing were found to be important indicators for improving calf production efficiency in Hanwoo farm. This study can be used as a research data to improve the reproductive rate of farmhouse sites through the survey on the breeding status of Hanwoo farmers.

• Journal of Embryo Transfer
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• v.24 no.1
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• pp.47-55
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• 2009

This study was performed to investigate the possibility of repeated superovulation treatment at interval from 27 days to 41 days in Hanwoo cattle and to compare with superovulation effect between doses of FSH 200 mg and FSH 400 mg. Different doses of FSH (200 mg or 400 mg) were injected at Day 8 after controlled internal drug release (CIDR) treatment for superovulation of Hanwoo donors following CIDR treatment (Day 8 after the estrus). Superovulation was repeated four times for one donor and number of corpus luteum (CL), number of embryos, number of transferable embryos and pregnancy rate after embryo transfer (ET) were investigated. 5 cows were used for each FSH treatment (10 cows in total). Average number of CL were $10.16{\pm}3.85$ and $11.56{\pm}2.35$ for the donors treated with FSH 200mg and FSH 400mg, respectively. Average number of embryos collected were $8.85{\pm}4.05$ and $8.30{\pm}1.73$ for the donors treated with FSH 200 mg and FSH 400 mg, respectively. Average number of transferable embryos were $5.48{\pm}2.45$ and $4.58{\pm}2.23$ for the donors treated with FSH 200 mg and FSH 400 mg, respectively. The pregnancy rate following ET with embryos collected from 200 mg FSH treated donors and 400 mg FSH treated donors were 61.9% and 53.8% respectively. The numbers of embryos tended to be decreased as the numbers of repeat of superovulation were elapsed. These results indicated that superovulation treatment by about a month to Hanwoo donors is usable and 200 mg of FSH is preferable for simple FSH treatment following CIDR treatment.

• Korean Journal of Animal Reproduction
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• v.10 no.1
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• pp.49-57
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• 1986

Studies were conducted to investigate the relationship between hormone leveles and nutritional levels for improving performance of Cheju native cattle. In June 1984 a trial was initiated using 8 Cheju native calves after weaning, fed at two su, pp.ementary feeding levels (NRC 100% and 70%). The body weight, breedng performence, change in progesterone level during pregnancy and estrus cycle were evaluated. Mean body weight at 6 months of age was 155kg when fed 100% NRC ration but it was only 137kg when heifers received the 70% NRC ration. At 10, 15 and 20 months of age the body weight was 66, 160 and 115kg, respectively, showing that heifers fed the standard ration gained weight rapidly (P<0.01). Average size of the lefe ovary in the standard group was 2.1${\times}$1.6cm and right ovary was 2.6${\times}$1.8cm. However in the restricted feeding group the ovaries were found to be smaller. Diameter of graffian follicles showed a similar tendency to ovarian size in the two groups. The first oestrus in the standard feeding group a, pp.ared at 14.6 months when body weight was 265kg. Age at first calving was on average 28.9 months at a body weight of 436kg. On the other hand when heifers were fed the restricted ration the first oestrus a, pp.ared at 23.0 months at a body weight of 250kg. Average age at first calving was 38.9 months which was 10 months later than the average in the standard feeding group (P<0.01). In standard feeding group the progesterone level was 2.0ng/ml at two weeks after pregnancy and gradully increased up to 4 weeks and peaked at 18 weeks. This peak (6.4-6.5ng/ml) was maintained up to 24 weeks when progesterone level decreased until it reached 2.1ng/ml at the end of pregnancy. In the restricted group progesterone level up to 16 weeks followed a similar pattern to the standard group but there was a tendency in the restricted group to have lower progesterone levels(P<0.01). The standard and srstricted groups showed similar patterns of progesterone concentration during the oestrus cylce. There were no statistically significant differences in progesterone levels between standard and restricted groups but there was variation between induvidual animals.

• Journal of Embryo Transfer
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• v.15 no.3
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• pp.255-261
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• 2000

The objective of this study was to improve the efficiency of bovine embryo transfer by transferring of Hanwoo embryos into Hanwoo or Holstein recipients. The cryopreserved or fresh in vitro produced(IVP) embryos were transferred into uterine horn contralaterally or ipsilaterally to the corpus luteum. The recipients were inseminated by artificially on the next day of estrus. The pregnancy was diagnosed by rectal palpation at 60∼90 days after transfer of the embryos. The pregnancy rate by transfer of one or two embryos was 78%(7/9) and 74%(31/42), respectively. The pregnancy rates according to the grade of corpus lutea of recipients was 75% (20/27) and 82.0%(18/22) at the grade of A and B, respectively. Ten(67.0%) of 15 Holstein recipients transferred with IVP Hanwoo embryos and 5(42.0%) of 12 Holstein recipients transferred with frozen IVP Hanwoo embryos were pregnant. The single and twin calving ratio in Hanwoos was 77.0%(10/13) and 23.0%(3.13) in the recipients transferred with IVP embryos and 64.0%(7/10) and 27.0%(3/10) in the recipients transferred with frozen IVP embryos, respectively. Twenty-four pregnant cows following transfer of IVP embryos, 21(88.0%) calved the normal calves, and 2(8.3%) aborted. When the frozen IVP embryos were transferred, 16 pregnant cows calved 14(88.0%) normal calves and 2(13.0%) aborted. In conclusion, when one or two IVP bovine embryos were transferred into recipients, the A and B grade of corpus luteum resulted in high pregnancy rates. For the production of twin calves, transfer of the IVP or frozen IVP embryos could be suitable.

• Proceedings of the KSAR Conference
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• 2001.03a
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• pp.7-7
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• 2001

Cryopreservation by vitrification of Hanwoo blastocysts derived from nuclear transfer with Hanwoo adult ear cell was compared with that of in vitro fertilized blastocysts. For vitrification, day 7 or day 8 blastocysts were serially exposed in glycerol (G) or/and ethylene glycol (EG) mixtures (10% G for 5 min, 10% G plus 20% EG for 5 min, and 25% G plus 25% EG for 30 sec) which was diluted in 10% FBS added D-PBS. Thawing of straw was carried out in air for 10 sec and then in water bath of $25^{\circ}C$ for 20 sec. The contents of the straw (0.2 ml) was expelled into a culture dish contained with 0.5M sucrose and 10% FBS(S-DPBS) by cutting the cotton plug and then the recovered embryos were put into fresh 0.25M and 0.125M S-DPBS for 30 sec, respectively, The embryos were transferred into D-PBS with 10% FBS for 5 min. and were cultured in a 10u1 droplet of co-culture environment (cumulus cell monolayer + 10% added CRl medium) for 24 h. In the result, survival rates were 88.9% and 85.4% for nuclear transfer embryos and in vitro fertilized embryos, respectively. After transfer of vitrified-thawed blastocysts produced from nuclear transfer, 4 of 5 total recipients did not return to the subsequent estrus cycle at 30 days. It is concluded that the Hanwoo blastocysts derived from nuclear transfer can be successfully cryopreserved using simple and efficient vitrification method.

• Asian-Australasian Journal of Animal Sciences
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• v.15 no.12
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• pp.1714-1718
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• 2002

Twelve prepubertal Karan Fries heifers (15 months, $167.7{\pm}13.5kg$) were divided into two equal groups. Group 1 was fed as per NRC requirements and group 2 was fed 20% more protein than group 1 heifers. The experimental feeding was continued until the onset of puberty in both the groups. Blood samples were collected at fortnightly intervals and analyzed for amino acids using HPLC. Group 1 and 2 heifers required $178.6{\pm}33.8$ and $152.8{\pm}33.2$ days of experimental feeding to exhibit first estrus resulting in total age at puberty as $639.4{\pm}27.3$ and $618.6{\pm}24.6$ days in the two groups respectively. The concentration of total amino acids averaged 4.40 and 4.89 mmol/l and those of non-essential amino acids (NEAA) was 2.32 and 2.49 mmol/l in groups 1 and 2, respectively. The concentration of plasma essential amino acids i.e. histidine, threonine, valine, methionine, isoleucine, leucine and phenylalanine were higher (p<0.01) in group 2 than group 1. Plasma concentration of large neutral amino acids (LNAA) was significantly higher in group 2 (1.28 mmol/l) than in group 1 (1.12 mmol/l). Increased levels of leucine, isoleucine and valine are implicated in increased follicular growth and development in prepubertal heifers and resulted in a 26 day earlier attainment of puberty by 26 days in an experimental period of six months in group 2 heifers. Increased concentrations of aspartate and tyrosine in group 2 heifers might be associated with the release of GnRH from the hypothalamus influencing LH release from anterior pituitary in such animals. It is therefore evident that increased availability of certain amino acids in heifers fed high protein diet might have led to early onset of puberty.

• Korean Journal of Animal Reproduction
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• v.10 no.1
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• pp.58-65
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• 1986

The results from a survery on the reproduction of a, pp.oximately 500 dairy cows raised in four different farming areas including Cheju city, Jochun, Hallim and Sung-up are summarized as follows; 1. At the occurrence of first estrus, the average age of the heifers was 15.1 months (330 Kg/B.W.), and at first conception 18.0 months (354Kg/B.W.). There was no regional differences. 2. The average interval of the estrous cycle was 20.4 days. And the repetition of esturs after unsuccessful insemination occurred on an average of 1.6 times, especially 2.0 times in the herds of the Jochun district. 3. The ages at the first calving were similar to the data recorded for the domestic mainland's herds at 27.1 months. But some regional differences were recognized within the four districts: i.e. 28.6 (Cheju city), 27.4(Jochun), 27.1(Hallim) and 25.3 months (Sung-up). There was no significant differences in sex ratio of newborns at 50.4%(♂) and 49.6%(♀). 4. The intervals of esturs recurrence and the following conception after calving were 80.0 and 95.8 days respectively. In the followsing conception intervals after cavling, 90.6-97.4 days were average for the herds of Cheju city and Jochun districts, 62.4-69.7 days for those of Hallim and Sung-up districts. 5. The average gestation period was 279.8 days, and was almost identical in the four districts. 6. The calving intervals was 377.7 days in average, and it was shorter than the data recorded for the domestic mainland's herds. Also it was reduce gradually according to the calving frequency.

• Journal of Embryo Transfer
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• v.12 no.1
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• pp.67-74
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• 1997

This study was carried out to increase the viability of bovine frozen4hawed in vitro produced (IVP) embryos and pregnancy rate by direct transfer method. Cumulus-oocyte complexes were aspirated from excised Hanwoo ovaries and matured in TGM 199 for 20~22 hours at 38.5$^{\circ}C$ in 2% $CO_2$ in air. Matured oocytes were fertilized with capacitated sperm for 6 hours and then co-cultured with cumulus cells for 9 days. 63% of the oocytes cultured was deaved and 29% out of them developed into blastocysts. Good or excellent grade of blastocysts on D 7 or 8 were frozen with 1.8M ethylene glycol as a cryoprotectant for direct transfer. Frozen embryos were thawed at 2$0^{\circ}C$ water for 10 sec following 4~5 second in air. For the survival assay of frozen4hawed lVP blastocysts, they were cultured in TCM 199 supplemented with 100$\mu$M $\beta$-mercaptoethanol and 20% FCS for 72 hours. The percentage of embryos developed to re-expanded or hatched after 72 hours culture was 95. 5 and 77.3%, respectively. When frozen-thawed Ivp embryos were transferred to 43 synchronized recipients by direct transfer method, eighteen recipients (41.8%) was pregnant. The highest pregnant was in naturafly synchronized recipients (71.4%), but induced estrus by using PRID(29.2%) and PGF$_2$$\alpha$(20.0%) was showed lower pregnancy rate. The pregnancy rate was higher in day 7 blastocysts(56.0%) than day 8 blastocysts(22.2%). (Key words: in vitro produced, blastocyst, frozen-thawed, direct transfer)