• Reproductive and Developmental Biology
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• 제34권3호
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• pp.201-205
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• 2010

The objective of this study was investigate the superovulation treatment and to relate concentrations of blood urea nitrogen(BUN) in Hanwoo donors. Thirty six, at random stages of the estrous cycle, received a CIDR. 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 On the 3th administration of FSH, CIDR was withdrawn and 25 mg $PGF_2a$ was administered.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 or 8 days after the 1st insemination. Cows with BUN <10, 11~18 and ${\geq}$19 mg/dl had return of estrus of 34.6, 30.5 and 30.4 days respectively. Return of estrus after superovulation treatment was not significantly lower for cows with blood urea nitrogen (BUN) above 10 mg/dl than for cows with BUN below 10 mg/dl. Cows with BUN <10, 11~18 and ${\geq}$19 mg/dl had number of transferable embryos of $3.2{\pm}1.2$, $5.4{\pm}1.9$ and $4.1{\pm}2.1$ respectively.

• Reproductive and Developmental Biology
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• 제34권3호
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• pp.181-184
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• 2010

Osmolarity of culture media is one of the most important factors affecting in vitro development. This study was conducted to investigate the DNA methylation status of Pre-1 and satellite sequence in pig nuclear transfer (pNT) embryos produced under different osmolarity culture conditions. Control group of pNT embryos was cultured in PZM-3 for six days. Other two treatment groups of pNT embryos were cultured in modified PZM-3 with 138 mM NaG or 0.05M sucrose (mPZM-3, 320 mOsmol) for two days, and then cultured in PZM-3 (270 mOsmol) for four days. Previous our studies have reported that pNT embryos cultured in both hypertonic media showed significantly higher blastocyst formation rate than that of control. The DNA methylation status of the satellite sequences in blastocyst was characterized using bisulfite-sequencing technology. The satellite region had a similar methylation pattern of in vivo blastocyst among two culture groups excepting the control group. Each level of methylation is that the satellite DNA moderately methylated (43.10% of PZM-3; 56.12% of NaCl; 55.06% of sucrose; 60.00% of in vivo embryos). As a result of the sequence of PRE-1, CpG methylation pattern was similar to three groups, including in vivo group. In case of the satellite DNA region, the osmolarity conditions were affected CpG DNA methylation status while PRE-1 sequence was not affected CpG DNA methylation in pNT blastocyst stage. These results indicate that the modification of osmolarity in a culture media may influence to spatially change of DNA methylation of repetitive sequence for pNT embryo development.

• 한국동물위생학회지
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• 제31권4호
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• pp.497-504
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• 2008

A cross-sectional study was undertaken to report prevalence and to identify risk factors of subclinical mastitis of dairy cattle in Sylhet district of Bangladesh. Among 325 dairy farms of the district 12 farms(3.7%) were selected conveniently for this study. All the dairy cows of the 12 farms were selected for sample collection. Fresh milk samples from each of the selected dairy cows were collected aseptically in separate sterilized test tube as RF, RH, LF and LH quarter of the udder. Rapid modified White Side Test(WST) was used to detect subclinical mastitis(SCM). Results of WST and data derived from filled in questionnaire were entered in Microsoft Excel 2003 and transferred to $STATA^{(R)}$, version 8.0/Intercooled(Stata Corporation, Texas, USA, 2003). The overall prevalence of SCM and its distribution in different categories of variables in cow and their exact binomial 95% confidence intervals were calculated in $STATA^{(R)}$. Simple bivariable associations among independent variables were investigated by $x^2$ test in $STATA^{(R)}$. Multiple logistic regression analysis with backward elimination method was used to identify risk factors of SCM. To identify significant variation in quarter SCM, linear regression analysis was performed after arcsine transformation of the data. The overall prevalence of SCM found in this study is 54%. Dairy cows with teat lesions had significantly increased SCM(OR=12342, P value=0.000, 95% CI=762, 199798) than others without teat lesions. The Holstein Friesian X Jersey X Sahiwal breed has significantly decreased(OR=0.18, p=0.03, 95% CI 0.04, 0.85) SCM than other breeds. The prevalence of SCM found in this study is in agreement with others. The injury in the teat increases the probability of getting infected with microbes and thereby mastitis. If the prevalence of teat lesion can be decreased the probability of subclinical mastitis will also be decreased. The negatively associated Holstein Friesian X Jersey X Sahiwall breed may help in planning mastitis control program if this finding can be validated by a more powerful case-control or cohort study design.

• 한국발생생물학회지:발생과생식
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• 제8권1호
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• pp.35-42
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• 2004

미성숙의 Germinal Vesicle(GV 단계에서 성숙한 Metaphase II(MII) 단계가 되는 난자성숙 과정은 핵과 세포질의 성숙을 통해 이루어지며, 이를 통해 수정과 배 발달을 할 수 있는 능력을 갖게 된다. GV 난자는 prophase I 단계에 arrest 되어 있다가 meiosis 과정을 거쳐 성숙한 MII로 되는데 이를 조절하는 기작에 대해서는 거의 알려져 있지 않다. 따라서 본 연구는 미성숙 난자와 성숙 난자간의 유전자 발현의 차이를 동정함으로써 난자성숙에 관여하는 유전인자를 밝히고자 하였다. GV와 MII 난자에서 mRNA를 정제한 후 ACP System을 이용하여 두 그룹간의 유전자 발현 차이를 분석하여 양적으로 서로 다르게 발현하거나 한쪽에서만 특이적으로 발현하는 유전자를 cloning하여 Sequencing과 BLAST search를 통해 분석하였다. ACP 1번부터 20번까지를 사용하여 32개의 유전자를 찾았으며 이중 26개가 기능적으로 알려진 유전자였다. Pscd2를 포함한 4개의 유전자는 GV에 특이적으로 발현하였고, PKD2와 CSN3를 포함하는 10개의 유전자는 GV에서 더 높게 발현하였으며 Diva를 포함하는 12개의 유전자는 MII에서 더 높게 발현하였다. 본 연구를 통해 분석된 모든 유전자는 난자에서의 발현은 보고되지 않은 것으로 ACP System을 통해 최초로 동정되었으며 특히 PKD-CSN Signaling pathway가 난자에서 발현함을 알 수 있었다. 본 연구는 난자 성숙 과정에서 서로 다르게 발현하는 유전인자를 성공적으로 동정하였으며 향후 이들의 기능을 연구함으로써 난자성숙 조절기전을 연구하는데 기여할 것으로 사료된다.

• Reproductive and Developmental Biology
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• 제35권3호
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• pp.349-353
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• 2011

The objective of this study was to investigate the relationship between estrous expression, body condition score (BCS), blood urea nitrogen (BUN) and number of transferable embryos for the purpose of improving reproductive performance in blood of Hanwoo donors. Sixty, at random stages of the estrous cycle, received a CIDR. 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 3th administration of FSH, CIDR was withdrawn and 25 mg PGF2 ${\alpha}$ 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 1nd insemination. Embryos were recovered 7 or 8 days after the 1st insemination. The estrous inducement rate and estrous expression rate were significantly lower for cows with BCS below 2.25 than for cows with BCS above 2.25. There was 50.0% of rate of mounting in cows with BCS below 2.25 whereas the rate of mounting was markedly increased in cows with BCS above 2.25 (94.1% and 89.5% for BCS 2.25~2.75 and BCS above 2.75 cows, respectively). Cows with BCS <2.25, 2.25~2.75 and ${\geq}$2.75 had number of transferable embryos of $4.5{\pm}0.7$, $5.9{\pm}1.8$ and $5.6{\pm}2.3$ respectively.

• Reproductive and Developmental Biology
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• 제35권3호
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• pp.335-339
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• 2011

The objective of this study was to investigate the relationship between concentration of urea nitrogen, glucose, cholesterol and number of transferable embryos for the purpose of improving reproductive performance in blood of Hanwoo donors. Fifty five, at random stages of the estrous cycle, received a CIDR. 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 3th 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 1nd insemination. Embryos were recovered 7 or 8 days after the 1st insemination. Cows with BUN < 10, 11~18 and ${\geq}$19 mg/dl had number of transferable embryos of $4.32{\pm}1.3$, $5.8{\pm}1.8$ and $4.7{\pm}2.1$ respectively. The mean numbers of total ova from < 10 and 10${\leq}$ of corpora lutea(CL) was 8.9 and 14.3, respectively. The number of transferable embryos differed between < 10${\leq}$ CL was 4.8 and 5.6, respectively.

• 한국응용곤충학회지
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• 제53권4호
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• pp.381-389
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• 2014

매실의 낙과 피해를 일으키는 복숭아씨살이좀벌은 경기도, 충청남북도, 전라남북도 지역의 매실과 살구, 그리고 중국산 복숭아 종자에서 확인되었다. 복숭아씨살이좀벌은 땅에 떨어진 핵과류의 핵 속에서 유충 상태로 월동하고, 노숙유충은 4월 중순까지, 번데기는 3월 하순에서 4월 하순까지 관찰되었으며, 성충은 4월 하순~5월 상순 사이에 90% 이상이 우화하는 것으로 조사되었다. 암컷 성충은 핵이 경화되기 이전 씨앗이 액상일 때 과실 씨앗의 외피 바로 밑에 산란하였으며, 산란관의 길이가 5 mm를 넘지 않아 산란이 가능한 매실의 크기는 직경이 2 cm를 넘지 않았다. 하나의 과실에 최대 5개의 알이 관찰되었으며, 알 기간은 2일을 넘지 않았다. 부화한 유충은 먼저 고형화되는 배로 이동하여 이를 섭식하면서 성장하는데 이 과정에서 유충들 사이의 경쟁으로 인해 한 마리의 유충만 생존하였다. 6월 상순경 피해를 받은 과실의 대부분이 부패증상을 보이며 낙과하였다. 땅에 떨어진 과실의 핵 속에서 이듬해 봄까지 노숙유충으로 월동하였다.

• Journal of Plant Biotechnology
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• 제34권4호
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• pp.375-380
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• 2007

국내 옥수수 순계주에서 Agrobacterium 공동배양으로 CP4 5-Enol-pyruvylshikimate-3-phosphate synthase (CP4 EPSPS) 유전자가 도입된 제초제저항성식물체를 개발하였다. 5개의 순계주 (HW1, KL103, HW3, HW4, HW7)의 미숙배를 Ubiquitin promoter-CP4 EPSPS 유전자와 CaMV35S promoter-nptII 유전자가 발현되도록 제조된 pCAMBIA2300 벡터를 C58C1 Agrobacterium에 형질전환하여 공동 배양하였다. 항생제로 paromomycin이 첨가된 배지에서 선발된 옥수수 형질전환체를 PCR, RT-PCR 및 Northern 분석을 통하여 유전자의 도입과 발현을 확인하였다. 또한 형질전환 식물체의 glyphosate 처리에 따른 shikimate 축적반응을 확인하였다. Paromomycin 저항성 캘러스 형성빈도는 옥수수 각 순계주 HW1, KL103, HW3, HW4, HW7에서 각각 0.37%, 0.03%, 2.20%, 2.37%, 0.81%로 나타났으며, PCR분석을 통하여 최종적으로 2개의 옥수수 순계주 (HW3, HW4)의 paromomycin 저항성 캘러스로부터 분화된 식물체에서 확인하였다. 이러한 형질전환체중에서 RT-PCR 및 Nothern blot 분석을 통하여 CP4 EPSPS 유전자가 발현되는 2개의 계통 (M266, M104) 을 선발하였고, shikimate 축적반응을 통하여 glyphosate에 대한 저항성을 갖는 계통 (M266)을 최종적으로 선발하였다. 이러한 결과는 국내 옥수수 순계주에서 제초제저항성을 갖는 옥수수 형질 전환체를 개발할 수 있음을 시사한다.

• 식물병연구
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• 제14권3호
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• pp.176-181
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• 2008

본 연구는 키다리병에 심하게 감염된 종자를 온탕침법과 prochloraz 유제로 종자소독 시 그 효과가 떨어지는 원인을 규명하기 위해 시행하였다. 키다리병이 발병한 포장에서 수확한 종자의 경우 병원균이 벼 종자의 배와 배유 안쪽까지 감염되어 있었다. $60^{\circ}C$에서 10분 동안 온탕 소독한 종자의 경우 종피 표면의 균사는 파괴되어 비정상적인 반면 배 주피 안쪽에 존재하는 균사는 정상적인 형태로 존재하였다. 포자는 종피 표면에서 비정상적인 포자와 정상적인 포자를 모두 확인하였다. prochloraz 유제로 $30^{\circ}C$ 24시간 침지소독 한 경우 종피의 균사는 완전히 파괴되었으나 포자는 정상과 비정상 포자 모두 확인할 수 있었다. 이러한 결과를 통해 온탕소독의 경우 종피 같은 물리적 한계로 인해 고열이 병원균에 전달되지 못하고, prochloraz의 경우 병원균 포자를 완전히 사멸시키기 못하기 때문에 종자소독 효과가 떨어짐을 알 수 있었다.

• 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.