• Asian-Australasian Journal of Animal Sciences
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• 제17권4호
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• pp.479-484
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• 2004

The study was conducted to assess the effect of milk production, parity, stage of lactation, season and individual milk components themselves on milk urea nitrogen (MUN) concentration and other milk components of 3,219 Holstein dairy cows in Korean dairy farms. The MUN concentrations in Korean dairy cows were estimated to 16.68$\pm$5.87 mg/dl. Milk yield was negatively correlated with fat and protein contents and somatic cell counts (SCC) in milk (p<0.01). The increasing MUN concentration has positive correlation with yield and fat content. By increasing somatic cell, milk yield was reduced and MUN level was increased. Cows in spring and winter produced more milk over 1.43 and 0.93 kg/day, respectively, than cows in summer (p<0.01). Milk urea nitrogen concentrations of milk produced in summer and fall were significantly lower (p<0.01) than those in spring and winter. Both MUN concentration and somatic cell counts were highest in winter. Milk yield was lower (p<0.01) in the first calving than other calving time and was tended to increase until the fifth parity and then decrease. Milk urea nitrogen and SCC were not related to parity of cows in this study. Milk yield and SCC were positively related to lactation period while MUN concentrations and milk fat and protein contents were negatively influenced by stage of lactation. In the present study, the relationship between MUN and reproduction of dairy cows was also investigated. Cow produced milk in high MUN concentrations (greater than 18 mg/dl) had more open days than cows in MUN concentrations less than 18 mg/dl. However, no significant difference between MUN concentration levels and frequency of artificial insemination was found in this study. It is suggested that although MUN values for nutritional management and measures of production or reproduction are used, non-nutritional factors should be considered.

• 대한수의학회지
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• 제24권1호
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• pp.91-98
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• 1984

A total of 466 foremilk from dairy farms in Chonnam district was examined for the subclinical mastitis over a period of one year, using a method of the electrical conductivities(EC); absolute conductivity(AC) and differential conductivity(DC) and quarter difference value(QD), in relation to the California mastitis test(CMT) and the direct somatic cell count(DSCC). The compatibility and efficiency rating between the EC values and the other screening tests was conducted. Obtained results are summarized as follows. 1. A linear relationship was found between the EC values and the CMT scores and direct somatic cell counts and it was found that electrical conductivity measurements were comparable with other screening tests for diagnosing animals with mastitis. 2. Compatibilities between the EC and CMT were 70.4% in AC, 74.6% in DC and 70.7% in QD, and that of the EC and DSCC were 53.0% in AC, 63.1% in DC and 53.2% in QD. On the other hand, relative efficiency ratings of Postle's equation between EC and CMT were 37.3% in AC, 26.5% in DC and 13.6% in QD, and that of the EC and DSCC were 33.1% in AC, 20.2% in DC and 11.9% in QD. 3. In the foremilk samples collected from damaged quarters determined by EC, the false positive rate wart higher than the false negative rate, and consequently tests of EC produced lower compatibility or efficiency rating scores. These tendencies suggested that any factors other than the mastitic condition influencing the EC values might be existed.

• 한국수정란이식학회지
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• 제22권4호
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• pp.235-243
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• 2007

Our goal was to examine the effects of early denudation on the enucleation efficiency and developmental competence of embryos following somatic cell nuclear transfer (SCNT) and parthenogenetic activation (PA). Oocytes were denuded following 30 h of in vitro maturation (IVM) and then cultured with (D+) or without (D-) their detached cumulus cells for additional $10{\sim}14$ h. Control oocytes were denuded after $40{\sim}44$ h of IVM. The size of the perivitelline space was larger at 40 h of IVM ($11.7{\sim}11.8{\mu}m$) than at 30 h ($8.9{\mu}m;$ p<0.01). The distances between the metaphase II (M II) plates and the polar bodies (PBs) were shorter in D+ ($19.4{\mu}m$) and D- oocytes ($18.9{\mu}m$) than in control oocytes ($25.5{\mu}m;$ p<0.01). Enucleation rates following blind aspiration at 40 h of IVM were higher (p<0.01) in D+ (92%) and D- oocytes (93%) compared to controls (82%). Early denudation did not affect oocyte maturation or the in vitro development of SCNT and PA embryos. When SCNT embryos from D+ oocytes were transferred to four gilts, pregnancy was established in two pigs, and one of them farrowed three live piglets. In conclusion, early denudation of oocytes at 30 h of IVM could improve the enucleation efficiency by maintaining the M II plate and the PB within close proximity and support the in vivo development of SCNT embryos to term.

• Animal Bioscience
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• 제34권4호
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• pp.499-505
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• 2021

Objective: The objective of this study was to evaluate the genetic behavior of a population of Holstein cattle in response to the variation of environmental temperature by analyzing the effects of genotype by environment interaction (GEI) through reaction norms for the somatic cell score (SCS). Methods: Data was collected for 67,206 primiparous cows from the database of the Paraná Holstein Breeders Association in Brazil, with the aim of evaluating the temperature effect, considered as an environmental variable, distinguished under six gradients, with the variation range found being 17℃ to 19.5℃, over the region. A reaction norm model was adopted utilizing the fourth order under the Legendre polynomials, using the mixed models of analysis by the restricted maximum likelihood method by the WOMBAT software. Additionally, the genetic behavior of the 15 most representative bulls was assessed, in response to the changes in the temperature gradient. Results: A mean score of 2.66 and a heritability variation from 0.17 to 0.23 was found in the regional temperature increase. The correlation between the environmental gradients proved to be higher than 0.80. Distinctive genetic behaviors were observed according to the increase in regional temperature, with an observed increase of up to 0.258 in the breeding values of some animals, as well as a reduction in the breeding of up to 0.793, with occasional reclassifications being observed as the temperature increased. Conclusion: Non-relevant GEI for SCS were observed in Holstein cattle herds of southern Brazil. Thus, the inclusion of the temperature effect in the model of genetic evaluation of SCS for the southern Brazilian Holstein breed is not required.

• Journal of Animal Science and Technology
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• 제45권5호
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• pp.739-748
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• 2003

우리나라 젖소 종모우의 국가단위 종축평가방법의 일환으로 이용되고 있는 산유형질 및 체세포지수의 305일 보정에 의한 유전평가 방법론에 대한 대안을 제시하기 위한 사전 연구의 일환으로써 검정일 기록을 이용하여 공변량 함수를 활용한 검정일 임의회귀모형에 대한 효율성을 검정하고 각 산유형질에 대한 비유일수별 유전적 변이를 추정하여 제시하고자 본 연구를 수행하였다. 본 연구에 이용된 자료는 농협중앙회에서 수행하고 있는 젖소 산유능력 검정사업에서 조사된 산유능력 검정기록과 한국종축개량협회에서 조사된 개체별 혈통자료를 이용하여 분석을 실시하였다. 분석에 이용된 자료는 혈통이 알려진 1산차 개체로써 검정일별 축군당 3기록 이상을 갖고 부모가 모두 알려진 개체만을 이용하였으며 한 유기동안에 개체별 7회 이상의 기록을 갖는 개체만을 선별하였다. 또한 자료분석의 효율성을 기하기 위하여 측정자료의 이상치는 제외하였으며 축군들간의 혈연적으로 연결되지 않은 축군 자료는 제외하였다. 분석모형은 단형질 개체모형으로써 검정년도-월-연령의 효과를 고정효과 공변량모형으로 적합시켰고 개체별 상가적 유전효과 및 영구환경효과를 개체내 검정일에 대한 임의 공변량 효과로 적합하여 모형을 설정, 유전분석을 실시하였다. 여기서 사용된 공변량은 검정일에 대하여 1차항 및 2차항을 포함하는 다항 공변량 함수를 활용하였다. 분석결과 유량에 대한 검정일별 유전력은 0.13~0.29으로 추정되었으며 검정일별 변이가 큰 것으로 추정되었다. 반면에 유지방율에 대한 유전력은 0.09~0.11으로 비교적 낮게 추정되었으며 유단백율에 대한 유전력은 0.12~0.19으로 추정되었다. 무지고형분율에 대한 유전력은 0.17~0.23으로 추정되었으며 체세포지수에 대한 유전력은 0.02~0.04으로 아주 낮게 추정되었다. 유량에 대한 유전적 변이가 비유일별로 다소 크게 추정되어 비유일에 따른 잔차 변이의 이질성(Heterogeneosity)이 클 것으로 사료되었으며 이러한 효과를 고려한 모형개발에 대한 연구가 필요할 것으로 사료되었으며 또한 다산차 기록을 활용한 분석 등에 대한 연구가 필요할 것으로 사료되었다. 국가단위 종축평가에 적합하기 위한 모형제시를 위해서는 본 분석에서 활용한 모형을 확장하여 분석된 종모우별 유전능력과 305d 사전 보정에 의한 평가에서의 순위와의 비교 연구가 선행되어야 할 것으로 사료되었다.

• 한국수정란이식학회지
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• 제31권3호
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• pp.179-183
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• 2016

Even though klotho deficiency in mice exhibits multiple aging-like phenotypes, studies using large animal models such as pigs, which have many similarities to humans, have been limited due to the absence of cell lines or animal models. The objective of this study was to generate homozygous klotho knockout porcine cell lines and cloned embryos. A CRISPR sgRNA specific for the klotho gene was designed and sgRNA (targeting exon 3 of klotho) and Cas9 RNPs were transfected into porcine fibroblasts. The transfected fibroblasts were then used for single cell colony formation and 9 single cell-derived colonies were established. In a T7 endonuclease I mutation assay, 5 colonies (#3, #4, #5, #7 and #9) were confirmed as mutated. These 5 colonies were subsequently analyzed by deep sequencing for determination of homozygous mutated colonies and 4 (#3, #4, #5 and #9) from 5 colonies contained homozygous modifications. Somatic cell nuclear transfer was performed to generate homozygous klotho knockout cloned embryos by using one homozygous mutation colony (#9); the cleavage and blastocyst formation rates were 72.0% and 8.3%, respectively. Two cloned embryos derived from a homozygous klotho knockout cell line (#9) were subjected to deep sequencing and they showed the same mutation pattern as the donor cell line. In conclusion, we produced homozygous klotho knockout porcine embryos cloned from genome-edited porcine fibroblasts.

• 한국수정란이식학회:학술대회논문집
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• 한국수정란이식학회 2002년도 국제심포지엄
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• pp.73-73
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• 2002

Nuclear transfer (NT) techniques have advanced in the last years, and cloned animals have been produced by using somatic cells in several species including pig. However, it is difficult that the nuclear transfer porcine embryos development to blastocyst stage overcoming the cell block in vitro. Abnormal segregation of chromosomes in nuclear transferred embryos on genome activation stage bring about embryo degeneration, abnormal blastocyst, delayed and low embryo development. Thus, we are evaluated that the correlations of the frequency of embryo developmental rates and chromosome aberration in NT and In viかo fertilization (IVF) derived embryo. We are used for ear-skin-fibroblast cell in NT. If only karyotyping of embryonic cells are chromosomally abnormal, they may difficultly remain undetected. Then, we evaluate the chromosome aberrations, fluorescent in situ hybridization (FISH) with porcine chromosome 1 submetacentric specific DNA probe were excuted. In normal diploid cell nucleus, two hybridization signal was detected. In contrast, abnormal cell figured one or three over signals. The developmental rates of NT and IVF embryos were 55% vs 63%, 32% vs 33% and 13% vs 17% in 2 cell, 8 cell and blastocyst, respectively. When looking at the types of chromosome aberration, the detection of aneuploidy at Day 3 on the embryo culture. The percentage of chromosome aneuploidy of NT and IVF at 4-cell stage 40.0%, 31.3%, respectively. This result indicate that chromosomal abnormalities are associated with low developmental rate in porcine NT embryo. It is also suggest that abnormal porcine embryos produced by NT associated with lower implantation rate, increase abortion rate and production of abnormal fetuses.

• Reproductive and Developmental Biology
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• 제34권3호
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• pp.127-134
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• 2010

This study was to investigate the effect of flavonoid treatment on in vitro development of bovine somatic cell nuclear transfer (SCNT) embryos, and their pregnancy and delivery rate after embryo transfer into recipient. In experiment 1, to optimize the flavonoid concentration, parthenogenetic day 2 ($\geq$ 2-cell) embryos were cultured in 0 (control), 1, 10 and $20\;{\mu}M$ flavonoid for 6 days. In the results, in vitro development rate was the highest in $10\;{\mu}M$ flavonoid group (57.1%) among treatment groups (control, 49.5%; $1\;{\mu}M$, 54.2%; $20\;{\mu}M$, 37.5%), and numbers of total and ICM cells were significantly (p<0.05) higher in $10\;{\mu}M$ flavonoid group than other groups. We found that $10\;{\mu}M$ flavonoid treatment can significantly (p<0.05) decrease the apoptotic index and derive high expression of anti-oxidant, anti-apoptotic, cell growth and development marker genes such as Mn-SOD, Survivin, Bax inhibitor, Glut-5, In-tau, compared to control group. In experiment 2, to produce the cloned Jeju Black Cattle, beef quality index grade 1 bull somatic cells were transferred into enucleated bovine MII oocytes and reconstructed embryos were cultured in $10\;{\mu}M$ flavonoid added medium. When the in vitro produced day 7 or 8 SCNT blastocysts were transferred into a number of recipients, $10\;{\mu}M$ flavonoid treatment group presented higher pregnancy rate (10.2%, 6/59) than control group (5.9%, 2/34). Total three cloned Jeju Black calves were born. Also, two cloned calves in $10\;{\mu}M$ flavonoid group were born and both were all healthy at present, while the one cloned calf born in control group was dead one month after birth. In addition, when the result of short tandem repeat marker analysis of each cloned calf was investigated, microsatellite loci of 11 numbers matched genotype between donor cell and cloned calf tissue. These results demonstrated that the flavonoid addition in culture medium may have beneficial effects on in vitro and in vivo developmental capacity of SCNT embryos and pregnancy rate.

• 생명과학회지
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• 제30권4호
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• pp.394-401
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• 2020

배아줄기세포는 만능세포이기 때문에 동물에게 주입되면 종양으로 발달할 수도 있다. 따라서 연구자들은 종양 형성으로부터 비교적 자유로운 성체세포로부터 세포 특이적 줄기세포(성체줄기세포)를 확보하는데 관심을 두고 있다. 성체줄기세포는 제한적으로 세포분열을 할 수 있고 지정된 특정 세포로만 발달할 수 있다. 포유동물에서 각 조직의 세포들은 자연적 생리조건하에서는 역분화 혹은 교차분화에 의해 성체줄기세포로 전환되지 않는다. 따라서 일본 연구자들에 의하여 2006년 성체세포의 리프로그램에 의한 유도만능줄기세포(iPSCs) 기술이 소개되어 성체줄기세포 연구의 새로운 장을 열었다. 비록 연구현장에서 iPSCs 기술이 폭 넓게 이용되지만, 리프로그램의 안정성뿐만 아니라 유전체에 외래유전자의 도입 등의 문제점도 있다. Reversine은 iPSCs 보다 2년 앞서 발견된 작은 화학적 합성 분자인 퓨린 유사체이다. Reversine은 분화된 세포를 리프로그램에 의한 역분화를 유도하여 다능성 줄기세포로 전환시킬 수 있으며, 적절한 분화조건하에서 다른 세포로 교차분화를 유도할 수도 있다. 따라서 reversine은 iPSCs가 가지고 있는 문제점을 극복하고 화학적인 방법을 이용하여 성체세포를 다능성 줄기세포로 전환시킬 수 있는 물질로 활용될 수 있다. Reversine이 백색지방세포를 갈색지방형세포(beige cell)로 전변시켜 열발산에 의한 에너지소비를 촉진함을 제시하여 항비만인자로서 그 가능성을 열어 놓았다. Reversine은 세포 역분화의 기능적 역할 이외에 항암 인자로서 또 다른 기능들이 보고되고 있어 앞으로 여러 분야에서 그 이용성이 기대되는 물질이다.

• 한국동물생명공학회지
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• 제35권3호
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• pp.258-264
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• 2020

This study investigated the effect of variation in the number of somatic-cell-cloned embryos and their developmental stage at transfer on pregnancy, as well as the influence of the estrus status of recipient pigs on in vivo development of cloned porcine embryos after embryo transfer. For somatic cell nuclear transfer (SCNT), fibroblast cells were obtained from a male porcine fetus. Recipient oocytes were collected from prepubertal gilts at a local abattoir and then cultured. After SCNT, reconstructed embryos of different numbers and developmental stages were transferred into recipient pigs. The developmental stage of the cloned embryos and the number of transferred embryos per surrogate showed no significant differences in terms of the resulting cloning efficiency. However, the pregnancy rate improved gradually as the number of transferred cloned embryos was increased from 100-150 or 151-200 to 201-300 per recipient. In pre-, peri-, and post-ovulation stages, pregnancy rates of 28.6%, 41.8%, and 67.6% and 16, 52, and 74 offspring were recorded, respectively. The number of cloned embryos and estrus status of the recipient pig at the time of transfer of the cloned embryo affect the efficiency of pig production; therefore, these variables should be particularly considered in order to increase the efficiency of somatic cell pig cloning.