• Clinical and Experimental Reproductive Medicine
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• v.11 no.2
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• pp.69-76
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• 1984

In order to obtain high proportion of Y sperm the semen was laid over 6%,10% and 20% bovine serum albumin. Separated sperm were stained with quinacrine mustard in order to see F - body which could be seen in human Y sperm. But we could'nt find F-body in the bull sperm. So sperm were compared with size of sperm head. As a result of observation separated sperm was small in size of length and width of sperm head as compared with control sperm. So it was found that the proportion of Y sperm showed a marked increase in separated layer. Then the higher albumin density was, the higher the proportion of Y sperm which had smaller head and faster motility was. But the higher albumin density was, the lower the recovery rate of sperm was. So it was hard to separate Y sperm in oligospermia.

• Korean Journal of Animal Reproduction
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• v.5 no.2
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• pp.35-42
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• 1981

A total 2,488 ejaculates during 4 years from 80 Korean native bulls over the age from 3 to 12 years of bull herds of Artificial Breeding Center, National Livestock Federatives Cooperation were collected and analyzed to study the effects of collection year, age of bulls, month of years, collection interval and ejaculation frequency per day on semen volume, sperm concentration, total sperm per ejaculate and sperm motility. 1. Semen volume, sperm concentration, total sperm per ejaculate and sperm motility index for the all ejaculates using least squares procedure averaged 5.73ml, 9.133${\times}$108/ml, 52.527${\times}$108 and 62.81, respectively. 2. Semen volume varied significantly with collection year, collection interval and ejaculation frequency per day(P<0.01), but effects of age of bulls and month of years on semen volume were not significant. Eight to 15 days collection interval showed the highest volume, and 1st ejaculate contained 5.6% more volume than 2nd ejaculate. 3. There were significant differences among collection years, months of years and ejeculation frequencies per day except collection intervals in sperm concentration per ml(P<0.05, P<0.01). Six to 8-year-old bulls was the highest concentration. Higher sperm concentration per ml was in April to July and lower month was October and December. Sperm concentration in 2nd ejaculate was higher than in 1st ejaculate. 4. Total sperm per ejaculate affected by all environmental factors studied(P<0.05, P<0.01). Age of bulls, collection interval and ejaculation frequency per day showed the highest total sperm was 6 to 8-year-old bulls, 8 to 15 days interval and 1st ejaculate, respectively. Higher total sperm per ejaculate was in April to July and lower total sperm was in September to December. 5. In sperm motility, there were significant differences among collection years, ages of bulls and collection intervals except months of years and ejaculation frequencies (P<0.01). Higher sperm motility was in 6 to 12-year-old bulls and in 5 to 7 days collection interval.

• Asian-Australasian Journal of Animal Sciences
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• v.20 no.1
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• pp.12-18
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• 2007

The objective of the study was to compare ranking of Sahiwal bulls selected on the basis of highest lactation milk yield of their dams with their estimated breeding values (EBVs) using an animal model. Data on 23,761 lactation milk yield records of 5,936 cows from five main Livestock Experiment Stations in Punjab province of Pakistan (1964-2004) were used for the study. At present the young A.I bulls are required to be from A-category bull-dams. Dams were categorized as A, B, C and D if they had highest lactation milk yield of ${\geq}$2,700, 2,250-2,699, 1,800-2,249 and <1,800 litres, respectively. The EBVs for lactation milk yield were estimated for all the animals using an individual animal model having fixed effect of herd-year and season of calving and random effect of animal. Fixed effect of parity and random effect of permanent environment were incorporated when multiple lactation were used. There were 396 young bulls used for semen collection and A.I during 1973-2004. However, progeny with lactation yields recorded, were available only for 91 bulls and dams could be traced for only 63 bulls. Overall lactation milk yield averaged 1,440.8 kg. Milk yield was 10% heritable with repeatability of 39%. Ranking bulls on highest lactation milk yield of their dams, the in-vogue criteria of selecting bulls, had a rank correlation of 0.167 (p<0.190) with ranking based on EBVs from animal model analysis. Bulls' EBVs for all lactations had rank correlation of 0.716 (p<0.001) with EBVs based on first lactation milk yield and 0.766 (p<0.001) with average EBVs of dam and sire (pedigree index). Ranking of bulls on highest lactation yield of their dams has no association with their ranking based on animal model evaluation. Young Sahiwal bulls should be selected on the basis of pedigree index instead of highest lactation yield of dams. This can help improve the genetic potential of the breed accruing to conservation and development efforts.

• Asian-Australasian Journal of Animal Sciences
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• v.31 no.7
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• pp.951-960
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• 2018

Hanwoo cattle are an important food source in Korea and their supply can have a major impact on meat availability for Korean consumers. The Hanwoo population was 1.8 million head in 2005 and gradually increased to 2.6 million in 2015. Per capita beef consumption has also increased, to 11.6 kg per year in 2015, and is expected to continue to increase. Because intramuscular fat percentage is a critical contributor to meat quality, Hanwoo cattle are fed a high-energy corn-based diet for long fattening periods. Long fed diet causes significant alterations in fat percentage in the loin muscle and other areas of the carcass. However, these long feeding periods increase feeding costs and beef prices. Recently, there has been increased Korean consumer demand for lean beef which has less fat, but is tender and priced more reasonably. These consumer demands on the Korean beef industry are driving differing beef production systems and also changes to the beef grading methodology. Korean government has made a significant investment to select bulls with favorable production traits using progeny testing. Progeny tested bull semen has been disseminated to all Hanwoo farmers. A beef traceability system has been employed for all cattle breeds in Korea since 2009. Hanwoo cattle are ear-marked with a 12-digit identification number from birth to slaughter. This number allows traceability of the management history of individual cattle, and also provides information to consumers. Traceability including management information such as herd, farm, year of birth, and carcass data can determine estimated breeding values of Hanwoo. For a sustainable Hanwoo industry, research scientists in Korea have attempted to develop feeds for efficient fattening periods and precision feeding systems based on genetic information for Hanwoo cattle. These initiatives aim to Korean consumer demands for beef and provide more precision management in beef production in Korea.

• Korean Journal of Animal Reproduction
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• v.26 no.3
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• pp.275-289
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• 2002

The objective of this study was to develop an in vitro assessment of sperm fertilizing capacity of bulls and investigate the factors influencing sperm function and characteristics of frozen-thawed bovine spermatozoa. in vitro fertilization (IVF), the evaluation of motility and normal morphology, HOST (hypoosmotic swelling test), Ca-ionophore induced acrosome reaction, luminol and lucigenin-dependent chemiluminescence for the measurement of reactive oxygen species (ROS), the measurement of malondialdehyde formation for the analysis of lipid peroxidation (LPO), and the evaluation of DNA fragmentation using the method of 747-mediated nick end labelling (TUNEL) by flow cytometry were performed in frozen-thawed bovine spermatozoa. Correlations between the rates of fertilization, blastocyst formation after IVF and the values of respective assays were investigated. 1. IVF rate and blastocyst formation rate averaged 64.4% and 34.3% for spermatozoa from high -fertility bull group and averaged 18.5% and 6.2% for spermatozoa from low-fertility bull group, respectively. There were significantly different between two bull groups. Sperm motility and percentage acrosome reaction averaged 79.0% and 66.2% for spermatozoa from high-fertility bull group and averaged 40.7% and 22.9% for spermatozoa from low-fertility bull group, respectivitely. There were not different between two bull groups. 2. Luminol depenent chemiluminescence, LPO and DNA fragementation averaged 6.4, 2.0 nmol and 2.6% from spermatozoa from high-fertility bull group and averaged 6.5, 3.1 nmol and 7.4% for spermatozoa from low-fertility bull group, respectively. There were significantly different between two bull groups. There was no significant difference in lucigenin dependent chemiluminescence between two bull groups. 3. Fertilization rate was positively correlated with motility and the rate of Ca-ionophore induced acrosome reaction, but negatively correlated with the frequency of luminol-dependent chemiluminescence, the rate of LPO, and the percentage of sperm with DNA fragmentation. There was no correlation between fertilization rate and the percentage of swollen spermatozoa, normal morphology, and the frequency of lucigenin-dependent chemiluminescence. 4. Blastocyst formation rate was positively correlated with the rate of Ca-ionophore induced acrosome reaction, but negatively correlated with the frequency of luminol-dependent chemiluminescence, the rate of LPO, and the percentage of sperm with DNA fragmentation. There was no correlation between blastocyst formation rate and motility, the percentage of swollen spermatozoa, normal morphology, and the frequency of lucigenin-dependent chemiluminescence. In conclusion, these data suggest that ROS significantly impact semen quality. The assays of this study may provide a basis fur improving in vitro assessment of sperm fertilizing capacity.

• Reproductive and Developmental Biology
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• v.29 no.3
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• pp.141-144
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• 2005

This experiment was to investigate whether the mitochondria function assessment can be used for the prediction of sperm fertility through examining the correlation between mitochondria fluoromicroscopic frequency of frozen-thawed eight Hanwoo bull semen using rhodamine123 (R123) and in vitro embryo development following fertilization. Individual sperm were stained in 5 ${\mu}g/mL$ R123-added calcium-free Sp-TALP for 30 min at 0 h, 6 h, 12 h and 24 h after thawing and examined their mid-piece under an epifluorescence microscope using 495 nm excitation filter (x1,000). Three replications were taken, and at least 300 sperm per individual were examined. When semen samples were separated into two groups (good and poor) by sperm motility and fluorescent frequencies at just after thawing, average fluorescent frequencies were remarkably reduced as time going (0 h; $53.29{\~}72.94\%$, 6 h; $21.40{\~}58.90\%$, 12 h; $8.26{\~}25.93\%$, 24 h; $1.00{\~}13.78\%$, irrespective of selected group, and there were no differences at 6 h or 12 h after thawing between selected groups but indicated significant difference at 24 h after thawing (p<0.05). In vitro fertilization rates in good and poor groups ranging $70.8{\~}77.8\%$ and $52.1{\~}84.5\%$, respectively, were not significantly different. However, in vitro development rates of the same groups ranging $25.7{\~}40.0\%$ and $12.9{\~}1.8\%$, respectively, were significant different (p<0.05). These results demonstrate that mitochondria fluoromicroscopic assessment of frozen-thawed bovine sperm may be used as a criterion to select more fertile sperm.

• Korean Journal of Veterinary Research
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• v.40 no.1
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• pp.187-195
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• 2000

The ejaculates from 67 HanWoo prove bull, bred in Livestock Improvement Main center of NLCF, were used to determine the correlation between the sperm motional characteristics and the pregnancy rate of artificial insemination(AI). The motional characteristics of sperm were analysed by Computer-assisted sperm analyser(CASA), thereafter inseminated equally 1,256 heads of cow regarding to parity, age, and live weight. There were no significant difference(p>0.05) in the pregnancy rate according to year from 1996 to 1998, but the LIN, ALH, STR, BCF, MAD and WOB of sperm in the year 1997, were highest pregnancy rate, were higher than those of sperm in the year 1998, were lowest pregnancy rate(p<0.05). The semen had no significant effect on pregnancy rate according to season(p>0.05). However spring, had a little higher pregnancy rate than that of autumn, were higher than autumn in VSL, VAP, LIN, ALH, BCF, MAD and WOB, but in DNM. The pregnancy rates of spring in the year 1996 and 1997 were higher than that of autumn in the year 1998(p<0.05). The spring in the year 1997, highest in pregnancy rate, were higher than the autumn in the year 1998 in VSL, VAP, LIN, STR, BCF, MAD and WOB, but in DNM(p<0.05). There were no the motion characteristic of sperm that was significant correlate with pregnancy rate of AI as the semen were analysed before artificial insemination and those, had some degree characteristics in motility, viability and abnormality, were used to AI. However there were a tendency that the higher the VSL, VAP, ALH, LIN, STR, BCF, MAD and WOB and the lower the DNM were, the higher the pregnancy rate of AI were.

• Proceedings of the Korean Society of Developmental Biology Conference
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• 2001.10a
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• pp.37-43
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• 2001

1. About fifty thousand of cattle embryos were transferred and 16000 ET-calves were born in 1999. Eighty percents of embryos were collected from Japanese Black beef donors and transferred to dairy Holstein heifers and cows. Since 1985, we have achieved in bovine in vitro fertilization using immature oocytes collected from ovaries of slaughterhouse. Now over 8000 embryos fertilized by Japanese Black bull, as Kitaguni 7~8 or Mitsufuku, famousbulls as high marbling score of progeny tests were sold to dairy farmers and transferred to their dairy cattle every year. 2. Embryo splitting for identical twins is demonstrated an useful tool to supply a bull for semen collection and a steer for beef performance test. According to the data of Dr. Hashiyada(2001), 296 pairs of split-half embryos were transferred to recipients and 98 gave births of 112 calves (23 pairs of identical twins and 66 singletons). 3. A blastomere-nuclear-transferred cloned calf was born in 1990 by a joint research with Drs. Tsunoda, National Institute of Animal Industry (NIAI) and Ushijima, Chiba Prefectural Farm Animal Center. The fruits of this technology were applied to the production of a calf from a cell of long-term-cultured inner cell mass (1988, Itoh et al, ZEN-NOH Central Research Institute for Feed and Livestock) and a cloned calf from three-successive-cloning (1997, Tsunoda et al.). According to the survey of MAFF of Japan, over 500 calves were born until this year and a glaf of them were already brought to the market for beef. 4. After the report of "Dolly", in February 1997, the first somatic cell clone female calves were born in July 1998 as the fruits of the joint research organized by Dr. Tsunoda in Kinki University (Kato et al, 2000). The male calves were born in August and September 1998 by the collaboration with NIAI and Kagoshima Prefecture. Then 244 calves, four pigs and a kid of goat were now born in 36 institutes of Japan. 5. Somatic cell cloning in farm animal production will bring us as effective reproductive method of elite-dairy- cows, super-cows and excellent bulls. The effect of making copy farm animal is also related to the reservation of genetic resources and re-creation of a male bull from a castrated steer of excellent marbling beef. Cloning of genetically modified animals is most promising to making pig organs transplant to people and providing protein drugs in milk of pig, goat and cattle. 6. Farm animal cloning is one of the most dreamful technologies of 21th century. It is necessary to develop this technology more efficient and stable as realistic technology of the farm animal production. We are making researches related to the best condition of donor cells for high productivity of cloning, genetic analysis of cloned animals, growth and performance abilities of clone cattle and pathological and genetical analysis of high rates of abortion and stillbirth of clone calves (about 30% of periparutum mortality). 7. It is requested in the report of Ministry of Health, labor and Welfare to make clear that carbon-copy cattle(somatic cell clone cattle) are safe and heathy for a commercial market since the somatic cell cloning is a completely new technology. Fattened beef steers (well-proved normal growth) and milking cows(shown a good fertility) are now provided for the assessment of food safety.

• Proceedings of the KSAR Conference
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• 2001.10a
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• pp.37-43
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• 2001

1. About fifty thousand of cattle embryos were transferred and 16000 ET-calves were born in 1999. Eighty percents of embryos were collected from Japanese Black beef donors and transferred to dairy Holstein heifers and cows. Since 1985, we have achieved in bovine in vitro fertilization using immature oocytes Collected from ovaries of slaughterhouse. Now over 8000 embryos fertilized by Japanese Black bull, as Kitaguni 7 -8 or Mitsufuku, famousbulls as high marbling score of progeny tests were sold to dairy farmers and transferred to their dairy cattle every year. 2. Embryo splitting for identical twins is demonstrated an useful tool to supply a bull for semen collection and a steer for beef performance test. According to the data of Dr.Hashiyada (2001), 296 pairs of split-half-embryos were transferred to recipients and 98 gave births of 112 calves (23 pairs of identical twins and 66 singletons). 3. A blastomere-nuclear-transferred cloned calf was born in 1990 by a joint research with Drs.Tsunoda, National Institute of Animal Industry (NIAI) and Ushijima, Chiba Prefectural Farm Animal Center. The fruits of this technology were applied to the production of a calf from a cell of long-term-cultured inner cell mass (1998, Itoh et al, ZEN-NOH Central Research Institute for Feed and Livestock) and a cloned calf from three-successive-cloning (1997, Tsunoda et al.). According to the survey of MAFF of Japan, over 500 calves were born until this year and a half of them were already brought to the market for beef. 4. After the report of "Dolly", in February 1997, the first somatic cell clone female calves were born in July 1998 as the fruits of the joint research organized by Dr. Tsunoda in Kinki University (Kato et al, 2000). The male calves were born in August and September 1998 by the collaboration with NIAI and Kagoshima Prefecture. Then 244 calves, four pigs and a kid of goat were now born in 36 institutes of Japan. 5. Somatic cell cloning in farm animal production will bring us an effective reproductive method of elite-dairy- cows, super-cows and excellent bulls. The effect of making copy farm animal is also related to the reservation of genetic resources and re-creation of a male bull from a castrated steer of excellent marbling beef. Cloning of genetically modified animals is most promising to making pig organs transplant to people and providing protein drugs in milk of pig, goat and cattle.

• Journal of Veterinary Clinics
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• v.7 no.2
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• pp.517-519
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• 1990

In vitro maturation and fertilization of oocytes collected from slaughtered bovine ovaries were investigated. Immature bovine extrafollicular oocytes were cultured for 24 hrs. in TCM 199 supplemented with fetal calf serum in a humidified CO$_2$ incubator. Fertilization in vitro was performed using frozen-awed bull semen which was treated by Ca Ionophore A23187. Fourty percentage of oocytes cultured had matured to the metaphase II ; There were-no effects of the concentration of fetal calf serum and of the addition of HEPES on the maturation rate. The mean proportions of in vitro fertilized eggs and of cleaved eggs were 23.1％ and 14.4％, respectively.