• 한국발생생물학회:학술대회논문집
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• 한국발생생물학회 2003년도 제3회 국제심포지움 및 학술대회
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• pp.108-108
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• 2003

The purpose of this is to investigate the effects of vitrification in open pulled straws (OPS) on in vitro survival of porcine embryos. Blastocysts were produced by in vitro fertilization of slaughterhouse-derived, in vitro matured oocytes with frozen-thawed boar semen, and subsequent culture on granulosa cell monolayer. After frozen-thawing, embryos were culture in NCSU-23 medium with 5 mM hypotaurine, 4 mg/$m\ell$ BSA and 10 ng/$m\ell$ for 48 hrs to survival tests. When blastocysts were frozen-thawed by OPS methods, the embryos with normal morphology were 32.1, 34.5 and 38.9 % in early blastocyst, blastocyst and expanded blastocyat stages. The rates of partial damaged embryos were significantly (P<0.05) higher in early biastocysts than expanded blastocysts. In another experiment, the embryos frozen by OPS methods were cultured for 48 hrs for survival and developmental rates in vitro. The proportions of embryos hatched were 11.8, 20.2 and 33.3% in embryos frozen-thawed at stages of early blastocyst, blastocyst and expanded embryos. On the other hand, The proportions of embryo with normal morphology after culture were 23.5, 25.0 and 33.3% in embryos frozen-thawed at stages of early blastocyst, blastocyst and expanded embryos. These finding indicate the possible broader application for OPS methods that this procedure described is relatively harmless, that it can be used for blastocysts of different developmental stages.

• 농업과학연구
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• 제37권2호
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• pp.267-270
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• 2010

This study was carried out to manufacture black charcoal board from domestic wood waste by using serum protein adhesive which is natural, environment-friendly and human-friendly. For the preparation of the serum protein adhesive, pig blood from slaughterhouse was centrifuged and serum was separated from corpuscles and concentrated to 30% by dry weight basis. The particle size of charcoal from domestic wood waste for this study was #6-60. Hot pressing schedule was $170^{\circ}C$ and 40kgf/$cm^2$ (1 min)-10kgf/$cm^2$ (2.5 min)-40kgf/$cm^2$ (5 min). The black charcoal board made by the addition of 13% serum protein adhesive on dry weight basis gave 41.76kgf/$cm^2$ of bending strength, 8.12kgf/$cm^2$ of internal bonding strength, and excellent gas adsorption and workability.

• 한국토양비료학회지
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• 제44권6호
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• pp.1245-1251
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• 2011

본 연구는 축산부문에서 주요한 가축종인 돼지의 사육과 정과 도축 가공과정에서 발생하는 양돈 바이오매스의 발생특성을 조사 분석하고, 전과정 평가 기법을 활용하여 물질(퇴 액비) 및 에너지 (바이오가스) 자원화 잠재량을 평가함으로써 지역단위 바이오매스 순환단지 조성을 위한 기초자료를 확립하고자 하였으며, 이를 위해 양돈 바이오매스의 발생 단계를 사양단계와 도축 가공단계로 구분하여 각각의 단계에서 발생하는 양돈바이오매스의 물질 및 에너지 자원화 잠재량을 평가하였다. 사양단계는 성장단계(사육기간, 평균체중)에 따라 자돈 (1~9주, 23.4 kg), 육성돈 1기 (10~15주, 50 kg), 육성돈 2기 (16~21주, 80 kg), 비육돈 (22~26주, 110 kg)의 단계로 분류하고 도축 가공단계에서 발생하는 혈액과 폐내장류, 장내 잔재물로 구분하여 생산량을 산정하여 양돈 바이오매스의 물질 및 에너지 자원 잠재량을 평가한 결과 돼지 1두에서 발생하는 바이오매스의 총량은 542.02 kg로 나타났다. 양돈 바이오매스는 분 $210.68kg\;head^{-1}$, 뇨 $315.78kg\;head^{-1}$가 발생하는 것으로 평가되었으며, 분뇨 발생량은 성장단계별로 자돈 14.2%, 육성돈 1기 19.6%, 육성돈 2기 30.9%, 비육돈 35.2%를 차지하는 것으로 나타났다. 양돈 바이오매스에서 기인하는 매탄 생산 잠재량은 $24.56Nm^3\;head^{-1}$이였으며, 사양 단계에서 기인하는 메탄 생산 잠재량이 92.9%를 차지하는 것으로 나타났다. BMP 시험에 의한 최대 메탄생산량은 $16.58Nm^3\;head^{-1}$로 나타나 매탄 생산 잠재량의 67.5%가 에너지로 전환 가능하였으며, 94.4%가 사양 단계에서 기인하는 것으로 나타났다.

• 한국발생생물학회:학술대회논문집
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• 한국발생생물학회 2001년도 발생공학 국제심포지움 및 학술대회 발표자료집
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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.

• Asian-Australasian Journal of Animal Sciences
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• 제21권10호
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• pp.1404-1410
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• 2008

This study was performed to clarify whether the variation of stress related heat shock protein 70 (HSP70) (GenBank X68213) gene was associated with the nuclear morphological change of in vitro maturation and in vitro capacitation in oocytes of pig ovaries obtained at the slaughterhouse. The nucleic acid substitution of C to G at the 483rd position was found out in HSP70 K1 (290-512) from X68213. The ovaries were categorized into CC, CG, and GG genotypes using polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) (BsiHKA I). After the second in vitro maturation of immature fresh oocytes, the relation of nuclear morphological change in oocytes with the genotype of HSP70 K1 gene was such that the MII ratios of the genotype GG and CG (46.93% and 42.20%, respectively) were significantly higher than that of the CC genotype (10.71%) (p<0.05). With respect to in vitro maturation of frozen-thawed oocytes by an open pulled straw (OPS) method, the percentage of oocytes matured to MII stage of the CG genotype showed a higher trend than CC and GG genotypes. After the in vitro maturation of immature fresh oocytes and frozen-thawed oocytes by the OPS method, the relation of the pronuclei change in oocytes matured in vitro with HSP70 genotype was assessed, and the result showed that the enlarged sperm heads (ESH) of matured fresh oocytes and frozen-thawed oocytes were 80.0% and 60.0% in the CC genotype, respectively. The CC genotype group had a significantly higher rate of ESH than the CG and the GG genotype group (p<0.05). The ratios of polyspermic invasion were not different among HSP70 of the three genotypes. It was considered that the rate of in vitro maturation of fertilized oocytes was expected to differ according to genotype of the stress related gene.

• 한국동물번식학회:학술대회논문집
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• 한국동물번식학회 2001년도 발생공학 국제심포지움 및 학술대회 발표자료집
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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.

• 한국수정란이식학회:학술대회논문집
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• 한국수정란이식학회 2002년도 국제심포지엄
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• pp.69-69
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• 2002

Transgenic animals production tools have been valuable for research and purpose. The current methods of gene transfer, microinjection and nuclear transfer, which are widely used in transgenic animal production, but all most methods has only had limited success in production of larger species. Here, we report the possibility of a sperm-mediated gene transfer method in porcine embryos. Oocytes were collected from ovaries harvested at a local slaughterhouse were matured in 500${mu}ell$ drops of TCM-199 under mineral oil at 38.5$^{\circ}C$ in a humidified atmosphere of 5%CO2 in air. After 42-43h of in vitro maturation oocytes were denuded. for sperm injection into the cytoplasm of the porcine oocytes, sperm suspension in NIM medium are subjected extraction with TritonX-100 before mixing with a green fluorescent gene (GFP). Sperm with Tritonx-100 were prepared by adding TritonX-100 to a final volume of 0.05% in the sperm suspension and mixing by trituration for 60s before two wishes in NIM medium at 2$^{\circ}C$. A(ter wishing, sperm were mixed with TritonX-100 at $25^{\circ}C$ followed by washes at 2$^{\circ}C$. Sperm were resuspended in ice cold NIM to a final volume of 400${mu}ell$ and 2-20ng/${mu}ell$ DNA were triturated on ice for 60s. All microinjection was performed in HEPES-buffered CZB medium at room temperature within 2h. After culture in NCSU-23 for 72h, percent of porcine embryos transfected GFP gene are 20.7%(6/29) in 20ng/${mu}ell$ sperm-DNA mixed group and other groups were 3.7 %(2/54)and 4.7%(3/67). These data suggests that sperm-mediated gene transfer method should be used to the production tool of transgenic pig efficiently.

• 한국발생생물학회:학술대회논문집
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• 한국발생생물학회 2003년도 제3회 국제심포지움 및 학술대회
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• pp.124-124
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• 2003

The successful development of embryos cloned by nuclear transfer (NT)have been dependent on a wide range of known factors including cell cycle of donor and recipient ooplast, oocyte quality, NT procedure and oocyte activation. The present study compared the development of cloned porcine embryos following different activation treatments. Cumulus-oocyte complexes (COCs) were aspirated from 26 mm follicles of slaughterhouse ovaries and cultured for 22 h in NCSU #23 medium supplemented with 10% porcine follicular fluid, 0.57 mM cysteine, 0.5 g/mL LH, 0.5 g/mL FSH and 10 ng/mL EGF. The COCs were further cultured for an additional 22 h in the same medium at $39{\cird}C$ in an atmosphere of 5% $CO_2$ in air, without hormonal supplements. Primary cultures of fibroblasts isolated from a female fetus on day 40 of gestation were established in DMEM + 15% FCS. For nuclear donation, cells at the 5th-6th passage were cultured in DMEM +0.5% FCS for 5 days in order to arrest the cells in G0/Gl. After enucleation, oocytes were reconstructed by transfer of donor cells and fusion with three DC pulses (1.4 KV/cm, 30 sec) in 0.28 M mannitol containing 0.01 mM $CaCl_2$ and 0.01 mM $MgCl_2$. Eggs were then divided into three treatment groups, control (without further treatment, Group 1), eggs cultured in 10 g/ml cycloheximide (CHX) for 5 h (Group 2), and eggs cultured in 1.9 mM 6-dimethylaminopurine (6-DMAP) for 5 h (Group 3). The eggs were then cultured in sets of 30 in 60 I drops of NCSU#23 supplemented with 4mg/ml BSA (essentially fatty acid free) until day 7 at $39{\circ}C$ in a humidified atmosphere of 5% $CO_2$. On day 4 the culture were fed by adding 20 I NCSU #23 supplemented with 10% FBS. Development rates into blastocysts were significantly higher (P<0.05) in Group 3 embryos compared to Group 1 controls ($27.6 \mu 2.7% vs. 20.1 \mu 4.1%$, respectively), but rates did not differ in Group 2 compared to control ($23.8 \mu 5.7%$). Total cell number in Group 3 blastocysts was however significantly higher (P<0.05) than in Groups 1 and 2 ($44.6 \mu 2.4 vs. 19.9 \mu 1.9 and 21.9 \mu 2.1$, respectively). These results suggest that 6-DMAP is more efficient than cycloheximide in the activation of electrically fused NT oocytes during in vitro production of cloned porcine embryos.

• 한국축산식품학회지
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• 제29권2호
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• pp.229-237
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• 2009

본 연구에서는 자돈(6.3 kg) 비육 출하시(110 kg)까지 식이유황의 급여가 돈육 품질특성에 미치는 영향을 조사하였다. 3원교잡종$(Landrace{\times}Yorkshire{\times}Duroc)$ 135두를 각 처리구 마다 45두씩(돈방당 15두)배치하여 158일간 사양시험을 실시하였다. 대조구는 식이유황 무첨가, 처리구 1은 300ppm, 처리구 2는 500ppm 첨가하여 급여하였다. 급여기간이 끝난 후 일괄적으로 함양도축장에서 도축하여 돈육의 등심부위를 wrap으로 함기포장하여 냉장온도$(4^{\circ}C)$에서 8일간 저장하면서 품질특성과 관련된 각 실험항목을 조사하였다. 지방산화는 저장기간 동안 대조구에 비하여 유황 급여 처리구가 낮은 지방산화를 보였으며, 저장기간에 따른 비교에서는 모든 처리구가 저장기간이 경과함에 따라 유의적으로 증가하였다(p<0.05). 관능적 특성 중 근내지방 함량과 전체적인 기호성은 유황 500ppm급여 처리구가 대조구와 유황 300ppm급여 처리구에 비하여 유의적으로 높은 값을 보였다. 저장기간의 경과에 따른 변화에서는 모든 처리구가 유의적인 차이가 없었다. 무기물 중 Na, Mg 및 Ca함량은 대조구에 비하여 유황 급여 수준이 증가할수록 함량이 유의적으로 감소하는 결과를 보였다(p<0.05). Fe, Cu 및 Zn 함량은 대조구에 비하여 유황 급여 처리구가 유의적으로 높은 함량을 보였다(p<0.05). S 함량은 대조구에 비하여 유황 급여수준이 증가할수록 유의적으로 증가하였다(p<0.05). 아미노산 함량은 대조구와 유황 급여 처리구간에 뚜렷한 경향이 없는 것으로 나타났다. 지방산 조성 중 palmitic, stearic과 oleic acid 함량은 대조구에 비하여 유황 급여 처리구가 유의적으로 높은 함량을 보였으며(p<0.05), 반면에 linoleic와 linolenic acid 함량은 유황 급여수준이 증가할수록 유의적으로 감소하였다(p<0.05). 포화지방산 함량은 유황 급여수준이 증가할수록 유의적으로 증가하였으며(p<0.05), 반면에 불포화지방산 함량은 유의적으로 감소하였다(p<0.05). 이상의 결과를 요약하면 식이유황 급여는 돈육 등심의 품질특성에 영향을 미치는 것으로 나타났다.