Chapter 1. Effect of OPU session periods on the efficiency of in vitro
embryo production in elite Korean native cow
Up-to-date artificial insemination (AI) using frozen sperm consider as the most widely using technology for improvement of Korean Native Cow (Hanwoo) embryo production. However...
Chapter 1. Effect of OPU session periods on the efficiency of in vitro
embryo production in elite Korean native cow
Up-to-date artificial insemination (AI) using frozen sperm consider as the most widely using technology for improvement of Korean Native Cow (Hanwoo) embryo production. However, it is time consuming, required at least 15~20 years to make more than 6 generations, and their offspring number is limited. To overcome such limitations, superovulation and in vitro fertilization have been developed. For superovulation, the number of produced embryos are not enough for commercialization and donor cows need rest period. This led to use of slaughterhouse ovary for in vitro fertilization, but it is impossible to repeat the collection from the same individual and it only can improve the genetic merits of offspring for one generation. Production of embryos using Ovum Pick-Up (OPU) technique, where oocytes can be repeatedly collected from living elite donor, might overcome these limitations. In this study, we investigated the possibility of using OPU technique from donors at different age and different session periods for mass-embryo-production. Oocytes were collected from 26 donor cows twice per week, 3 - 4 months per year, between 2013 and 2016. Results showed that, the average number of embryo produced in first year used donor was significantly higher than that in second year used donor (3.89 ± 2.85 vs 3.29 ± 2.70), however, there was no significant difference between third year used donor (3.51 ± 3.32) and other groups. Taken together, our data showed that repeated using of donor up to three years is possible for in vitro embryo mass-production. Moreover, OPU can be used as suitable embryo producing technique for livestock breed improvement.
Chapter 2. Effect of serum and serum free media on the developmental
competence of OPU derived bovine IVP embryo
Embryos produced with serum show alterations in their ultrastructure, impaired compaction, abnormal blastulation, aberrant mRNA expression profiles and large calf syndrome with greater incidences of stillbirths and deaths after birth. The aim of the present study was to describe in vitro embryo production by analyzing embryo production, fetal production and pregnancy rate in serum-free medium. The OPU-IVP data used in this study from 2016. Sixteen Korean native cows (Hanwoo) approximately, which belonged to the Institute of Agriculture and Life Science, Gyeongsang National University, were used. Experiment group is 2group. Donor is equivalent. Serum group for 2016 march to July and non-serum group for 2016 September to December. The recovered cumulus-oocyte complexes were morphologically classified into four grades based on the number of cumulus cells layers and homogeneity of the cytoplasm. Oocytes number for session was greater in serum groups than those in non-serum groups (29.61±0.63 vs. 15.6±0.62; P 0.05), but in grade 3+4 mean between greater in CR1aa group than that in non-serum group (7.31±7.64 vs.5.60±6.29; P < 0.05). Embryo cleaved competence rate in Non-serum groups was higher than those in serum (62.1% vs. 58.3; P < 0.05; Table 2). But, blastocyst developmental rate in serum was no difference than that in Non-serum (33.1% vs. 43.5%; P < 0.05). Embryo transfer for 986 recipients. Pregnancy rate were result of serum was no difference than that in Non-serum (54.6% vs. 56.3%; P < 0.05). In conclusion, the serum-free system we developed to produce in vitro bovine embryos meets the developmental and qualitative requirements for largescale commercial use.
Chapter 3. Analysis of offspring to Korean native cow embryo transfer
from OPU derived
This study is based on the result of slaughter by offspring of OPU derived embryo produce to check efficiency of embyro transfer and their offspring ability, then is able to apply its base data to embryo transfer. Retrieved data of offspring by embryo transfer were collected from 2015 to 2019. The number of posted offspring is 1145, and it's comprised of 983 of bulls and 162 of cow. Average months of slaughter is 29.4 months for bull and 43.0 months for cow. Grades of offspring is 393 (34.32%) for grade 1++, 455 (39.74%) for grade 1+, 186 (16.24%) of grade 1, 76 (6.64%) for grade 2, 27 (2.36%) for grade 3, meat quantity is 312 (27.25%) for grade A, 503 (43.93%) for grade B, 322 (28.12%) for grade C, and 8 (0.7%) for grade D. Meat quality of bull is 366 (37.23%) for grade 1++, 412 (41.91%) for grade 1+, 143 (14.55%) for grade 1, 42 (4.27%) for grade 2, 18 (1.83%) for grade 3, and its quantity is 277 (28.18%) for grade A, 430 (43.74%) for grade B, 274 (27.87%) for grade C, 2 (0.20%). Grade 1++ and 1+ form 79.14% of all. Animal products grading service is organized by carcass weight, backfat thickness, eye muscle area, marbling score, carcass weight of bull is 484.17±60.96, and cow is 386.75±69.96, eye muscle area of bull is 103.09±15.96 and cow is 88.90±17.47, backfat thickness of bull is 13.27±7.49, and cow is 14.50±8.10, and marbling score of bull is 6.71±1.85, cow is 5.17±2.24. Results of bull animal products grading service by years; carcass weight of 2015 is 429.56±82.91, 2016 is 482.99±46.80, 2017 is 462.87±59.76, 2018 is 495.98±55.36, and 2019 is 497.29±63.69, eye muscle area of 2015 is 89.93±15.43, 2016 is 94.32±17.55, 2017 is 102.03±16.72, 2018 is 106.22±14.17, and 2019 is 105.91±14.98, backfat thickness of 2015 is 11.67±5.13, 2016 is 16.36±14.49, 2017 is 13.40±8.24, 2018 is 12.86±4.85, and 2019 is 12.51±4.39. And marbling score of 2015 is 6.11±2.26, 2016 is 6.63±1.94, 2017 is 6.55±1.74, 2018 is 6.82±1.87, and 2019 is 6.83±1.83. Average months of slaughter of 2015 is 26.7, 2016 is 29.8, 2017 is 29.5, 2018 is 30, and 2019 is 28.9. Offspring grades by sperm, which 5 sperms are checked, (also) show differences. Donors also make its offspring grade different. Differences by sperm and donor are checked, and it is in the correlation between each. Compared animal products grading service of first generation and second generation, first generation shows more carcass weight, but second generation shows higher score of backfat thickness, eye muscle area and marbling score. Based on all these results, efficiency of embryo transfer is high, and this study will be help development of Hanwoo industry to be base date.
Chapter 1. Effect of OPU session periods on the efficiency of in vitro
embryo production in elite Korean native cow
Up-to-date artificial insemination (AI) using frozen sperm consider as the most widely using technology for improvement of Korean Native Cow (Hanwoo) embryo production. However, it is time consuming, required at least 15~20 years to make more than 6 generations, and their offspring number is limited. To overcome such limitations, superovulation and in vitro fertilization have been developed. For superovulation, the number of produced embryos are not enough for commercialization and donor cows need rest period. This led to use of slaughterhouse ovary for in vitro fertilization, but it is impossible to repeat the collection from the same individual and it only can improve the genetic merits of offspring for one generation. Production of embryos using Ovum Pick-Up (OPU) technique, where oocytes can be repeatedly collected from living elite donor, might overcome these limitations. In this study, we investigated the possibility of using OPU technique from donors at different age and different session periods for mass-embryo-production. Oocytes were collected from 26 donor cows twice per week, 3 - 4 months per year, between 2013 and 2016. Results showed that, the average number of embryo produced in first year used donor was significantly higher than that in second year used donor (3.89 ± 2.85 vs 3.29 ± 2.70), however, there was no significant difference between third year used donor (3.51 ± 3.32) and other groups. Taken together, our data showed that repeated using of donor up to three years is possible for in vitro embryo mass-production. Moreover, OPU can be used as suitable embryo producing technique for livestock breed improvement.
Chapter 2. Effect of serum and serum free media on the developmental
competence of OPU derived bovine IVP embryo
Embryos produced with serum show alterations in their ultrastructure, impaired compaction, abnormal blastulation, aberrant mRNA expression profiles and large calf syndrome with greater incidences of stillbirths and deaths after birth. The aim of the present study was to describe in vitro embryo production by analyzing embryo production, fetal production and pregnancy rate in serum-free medium. The OPU-IVP data used in this study from 2016. Sixteen Korean native cows (Hanwoo) approximately, which belonged to the Institute of Agriculture and Life Science, Gyeongsang National University, were used. Experiment group is 2group. Donor is equivalent. Serum group for 2016 march to July and non-serum group for 2016 September to December. The recovered cumulus-oocyte complexes were morphologically classified into four grades based on the number of cumulus cells layers and homogeneity of the cytoplasm. Oocytes number for session was greater in serum groups than those in non-serum groups (29.61±0.63 vs. 15.6±0.62; P 0.05), but in grade 3+4 mean between greater in CR1aa group than that in non-serum group (7.31±7.64 vs.5.60±6.29; P < 0.05). Embryo cleaved competence rate in Non-serum groups was higher than those in serum (62.1% vs. 58.3; P < 0.05; Table 2). But, blastocyst developmental rate in serum was no difference than that in Non-serum (33.1% vs. 43.5%; P < 0.05). Embryo transfer for 986 recipients. Pregnancy rate were result of serum was no difference than that in Non-serum (54.6% vs. 56.3%; P < 0.05). In conclusion, the serum-free system we developed to produce in vitro bovine embryos meets the developmental and qualitative requirements for largescale commercial use.
Chapter 3. Analysis of offspring to Korean native cow embryo transfer
from OPU derived
This study is based on the result of slaughter by offspring of OPU derived embryo produce to check efficiency of embyro transfer and their offspring ability, then is able to apply its base data to embryo transfer. Retrieved data of offspring by embryo transfer were collected from 2015 to 2019. The number of posted offspring is 1145, and it's comprised of 983 of bulls and 162 of cow. Average months of slaughter is 29.4 months for bull and 43.0 months for cow. Grades of offspring is 393 (34.32%) for grade 1++, 455 (39.74%) for grade 1+, 186 (16.24%) of grade 1, 76 (6.64%) for grade 2, 27 (2.36%) for grade 3, meat quantity is 312 (27.25%) for grade A, 503 (43.93%) for grade B, 322 (28.12%) for grade C, and 8 (0.7%) for grade D. Meat quality of bull is 366 (37.23%) for grade 1++, 412 (41.91%) for grade 1+, 143 (14.55%) for grade 1, 42 (4.27%) for grade 2, 18 (1.83%) for grade 3, and its quantity is 277 (28.18%) for grade A, 430 (43.74%) for grade B, 274 (27.87%) for grade C, 2 (0.20%). Grade 1++ and 1+ form 79.14% of all. Animal products grading service is organized by carcass weight, backfat thickness, eye muscle area, marbling score, carcass weight of bull is 484.17±60.96, and cow is 386.75±69.96, eye muscle area of bull is 103.09±15.96 and cow is 88.90±17.47, backfat thickness of bull is 13.27±7.49, and cow is 14.50±8.10, and marbling score of bull is 6.71±1.85, cow is 5.17±2.24. Results of bull animal products grading service by years; carcass weight of 2015 is 429.56±82.91, 2016 is 482.99±46.80, 2017 is 462.87±59.76, 2018 is 495.98±55.36, and 2019 is 497.29±63.69, eye muscle area of 2015 is 89.93±15.43, 2016 is 94.32±17.55, 2017 is 102.03±16.72, 2018 is 106.22±14.17, and 2019 is 105.91±14.98, backfat thickness of 2015 is 11.67±5.13, 2016 is 16.36±14.49, 2017 is 13.40±8.24, 2018 is 12.86±4.85, and 2019 is 12.51±4.39. And marbling score of 2015 is 6.11±2.26, 2016 is 6.63±1.94, 2017 is 6.55±1.74, 2018 is 6.82±1.87, and 2019 is 6.83±1.83. Average months of slaughter of 2015 is 26.7, 2016 is 29.8, 2017 is 29.5, 2018 is 30, and 2019 is 28.9. Offspring grades by sperm, which 5 sperms are checked, (also) show differences. Donors also make its offspring grade different. Differences by sperm and donor are checked, and it is in the correlation between each. Compared animal products grading service of first generation and second generation, first generation shows more carcass weight, but second generation shows higher score of backfat thickness, eye muscle area and marbling score. Based on all these results, efficiency of embryo transfer is high, and this study will be help development of Hanwoo industry to be base date.
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