• Journal of Embryo Transfer
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• v.24 no.1
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• pp.33-37
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• 2009

Multiple ovulation and embryo transfer (MOET) has the potential to increase the rates of genetic improvement in cattle. Thus this study was performed to investigate several factors influencing in vivo embryo production in Holstein cattle under field conditions. The donors were superovulated with Folltropin-V and $PGF_2{\alpha}$ combination method. From Day 10 onward, donors were superovulated by i.m., twice daily, administration of 400mg Folltropin-V given in a series of decreasing doses over a 4-day period: on the first day, 3.5ml; on the second day, 3.0ml; on the third day, 2.0ml; and on the fourth day, 1.5ml (20ml in total, equivalent to 400mg of NIH-FSH-P1). Estrus was induced by i.m. administration of 25mg prostaglandin $F_2{\alpha}$ on the sixth and seventh of FSH treatment. Estrus detection was performed twice daily beginning 24h after the first prostaglandin $F_2{\alpha}$ injection. Donor cows were artificially inseminated 12 and 24 h after first standing estrus with semen from a proven Holstein sire. Embryos used in this study were recovered Day 7.5 of the cycle (Day 0: first standing estrus). From 195 superovulated dairy cows, 2,104 eggs were recovered, of which 1,172 were classified as transferable embryos based on morphological evaluation of quality. The results are summarized as follows: 1. The numbers of recovered and transferable embryos did not significantly differ among the capacity of milk production that were < 10,000kg/305days (group 1), $10,000{\sim}12,000\;kg$/305days (group 2) or > 12,000kg/305 days (group 3) (p>0.05, Table 1). 2. No differences in the numbers of recovered and transferable embryos were found among the donor's postparient days (p>0.05, Table 2). 3. Also, the numbers of recovered and transferable embryos of each superovulation seasons did not significantly differ among the four groups (p>0.05, Table 3).

• Korean Journal of Animal Reproduction
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• v.27 no.3
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• pp.259-268
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• 2003

Lactadherin (formerly known as BA46), a major glycoprotein of the human milk fat globule membrane, is abundant in human breast milk and breast carcinoma cells and is known to prevent symptomatic rotavirus infections. In this study, we tried to transfer the human lactadherin gene to the Chinese Hamster Ovary (CHO) cells using retrovirus vector system and tested inducible expression of the gene under the tetracycline-controllable promoter. At first, tetracycline-mediated inducibility was tested using E.coli LacZ marker gene. NIH3T3 cells co-infected with RevTet-On and RevTRE-LacZ retrovirus vectors showed that the cells responded to doxycycline (a derivative of tetracycline) in a dose-dependent manner, and prominent induction of the lacZ gene expression was observed from 1 $\mu\textrm{g}$/ml of doxycycline concentration. Based on the results of the pilot experiment, inductional expression of the human lactadherin gene was conducted using RevTet-On and RevTRE-Ltd retrovirus vectors. Analysis with the RT-PCR demonstrated successful inductional expression of the lactadherin gene in the Chinese Hamster Ovary (CHO) cells. Considering that constitutive overexpression of the exogenous genes in the target cells causes serious physiological imbalance, the results obtained in this study will be very useful especially in the studies of gene therapy and transgenic animal production.

• Asian-Australasian Journal of Animal Sciences
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• v.24 no.9
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• pp.1318-1328
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• 2011

The tropics is not an ideal location for calf rearing as the high temperatures and humidities introduce many potential disease problems to milk fed calves. In addition, the type of dairy farming (generally poorly resourced small holder farming) and the general lack of awareness of the long term implications of poorly reared stock do not encourage farmers to pay close attention to their calf and heifer rearing systems. Surveys of calf rearing systems in Asia, tropical Africa and South America highlight the high calf and heifer mortalities. A range of 15 to 25% pre-weaning calf mortality is typical on many tropical dairy farms. It is often as high as 50%, indicating very poor calf management. This contrasts with US findings of less than 8% mortality from birth to 6 months while surveys of Australian farmers report only 3% losses. Simple extension programs on farms in Sri Lanka and Kenya have drastically reduced calf mortalities and improved pre-weaning growth rates. Improved management strategies leading to lower calving intervals, higher calving rates, reduced still born and pre-weaned calf mortalities and fewer non pregnant heifers can supply many more dairy herd replacements than currently occurs. Such strategies can increase the number of replacement heifer calves in the herd from 15 to over 35%, thus allowing farmers to increase their herd sizes through natural increases. Simple management procedures such as ensuring adequate intake of good quality colostrum within the first 12 hours of life, housing and good hygiene to minimise disease transfer, providing clean drinking water, developing appropriate feeding protocols to encourage early rumen development and paying closer attention to climate control and animal health can all lead to improved calf vigour and performance. Good record keeping is also important so farmers can more easily identify susceptible calves and quickly treat potential problems.

• Journal of Embryo Transfer
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• v.19 no.1
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• pp.53-59
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• 2004

The environmental impact of nutrient waste from agriculture has become an area of concern as ways to produce more food and of offspring. In dairy cattle, as the genetic capacity for milk production has increased, decreased fertility has become a severe problem by feeding high dietary protein, resulting in high concentration of blood urea nitrogen (BUN). There are numerous reports on BUN which is associated with reduced conception rates and closely related to body condition score (BCS) in lactating cattle, but not in Korean native cattle. This study was therefore performed to investigate the relation of BUN to both BCS and conception rates in Korean cattle. A total of 400 female Korean cattle (2∼5 years) in Daewanryong were used for this experiment. Feeding condition divided into two groups depending on grazing and darn feeding period. In grazing period, the mixture of Timothy, Orchard grass and Tall fescue, and concentrates (2.5 kg/day) was fed whereas hay (6.5 kg/day) and corn silage (20 kg/day) were fed while barn feeding period. Average BUN concentration at grazing and barn feeding were 7.39$\pm$2.65 mg/㎗ and 12.36$\pm$2.92 mg/㎗, respectively. During grazing period, high rates of pregnancy showed at 4∼8 mg/㎗ BUN concentration. In barn feeding period, 66%, of cattle were in pregnant at 8∼l4 mg/㎗ BUN concentration. The BCS for obtaining high rate of pregnancy raged at 2.5∼3.5. However, BUN did not directly relate to BCS in Korean cattle.

• Journal of Embryo Transfer
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• v.32 no.3
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• pp.227-233
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• 2017

Pig has been known to be one of the most feasible animals as a bioreactor to produce pharmaceuticals in milk and as a mediator in xenotransplantation research. Previously, we generated transgenic pigs for both purposes, which were expressing Factor 8, vWF, hTPA, and hEPO in milk, along with expression of MCP at GalT gene locus ($GalT^{-MCP/-MCP}$) as well as expressing MCP at GalT gene loci with CD73 expression ($GalT^{-MCP/+}/CD73$). In this study, we performed comparative analyses of sperm parameters between wild type male (WT) pig and those transgenic males to examine the effects of transgenes integrated into the pigs on motility, morphology, viability, and acrosome integrity of the spermatozoa. Our results showed that the rates of actively motile spermatozoa of WT, Factor 8, vWF, hTPA, hEPO, $GalT^{-MCP/+}/CD73$, and $GalT^{-MCP/-MCP}$ pigs were 85.0%, 83.3%, 82.5%, 83.3%, 82.5%, 77.5%, and 78.7%, respectively. Whereas, the rates of morphologically normal spermatozoa of WT, Factor 8, vWF, hTPA, hEPO, $GalT^{-MCP/+}/CD73$, and $GalT^{-MCP/-MCP}$ pigs were 90.0%, 80.0%, 80.0%, 83.3%, 85.0%, 91.8%, and 80.8%, respectively. In addition, the viability in spermatozoa of WT, Factor 8, vWF, hTPA, hEPO, $GalT^{-MCP/+}/CD73$, and $GalT^{-MCP/-MCP}$ pigs were 93.9%, 82.4%, 89.9%, 83.9%, 87.4%, 92.8%, and 83.6%, respectively. The rates of spermatozoa with normal acrosome integrity in WT, Factor 8, vWF, hTPA, hEPO, $GalT^{-MCP/+}/CD73$, and $GalT^{-MCP/-MCP}$ pigs were 98.1%, 98.6%, 98.6%, 98.7%, 98.1%, 99.5%, and 95.1%, respectively. There were no significant differences in motility, morphology, viability, and acrosome integrity of the spermatozoa among WT, Factor 8, vWF, hTPA, and hEPO, $GalT^{-MCP/+}/CD73$, and $GalT^{-MCP/-MCP}$ pigs. These mean that neither random integration nor targeted integration of the transgene into chromosome of pig effect on characteristics of spermatozoa. Ultimately, the transgenic male pigs subjected in this study could apply to propagate their progenies for production of human therapeutic proteins and advancing the xenotransplantation research.

• Journal of Embryo Transfer
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• v.20 no.3
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• pp.279-287
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• 2005

This study was carried out to investigate resumption of ovarian cyclicity and effect of BCS on estrous response by treatment of $PGF_2a+PGF_2a+CIDR$ program on day 40 postpartum in lactating dairy cow. First $PGF_2a$ was given on day 40 postpartum, second $PGF_2a$ was given 14 days apart to cows not-responded to 1st $PGF_2a$ and then CIDR was inserted for 7 days after 5 days in cows not-responded to 2nd $PGF_2a$. The $42.9\%$ of the cows showed more than 1 ng/mL milk progesterone concentration within 10 to 30 days postpartum. About $19\%$ of the cows exhibited more than 1 ng/mL milk pro-gesterone concentration between 31 to 50 days postpartum. However $38.1\%$ of the cows have not shown more than 1 ng/mL milk progesterone up to 50 days postpartum. Estrous response to the treatment of 1 st $PGF_2a$ and 2nd $PGF_2a$ was $47.5\%$ and $52.4\%$, respectively. Combination of 1 st $PGF_2a$ and 2nd $PGF_2a$ was $75\%$ and combination of 1st $PGF_2a$+2nd $PGF_2a$+CIDR was $87.5\%$. Estrous response to the treatment of $PGF_2a+PGF_2a$ program was $61.5\%$ in cows with less than 2.50 BCS and $81.5\%$ in cows with 2.75${\~}$3.50 BCS. Estrous response to the treatment of CIDR was $40\%$ in cows with less than 2.50 BCS and $80\%$ in cows with 2.75${\~}$3.50 BCS. Estrous response to the treatment of PPC on day 40 postpartum was $76.9\%$ in cows with less than 2.50 BCS and $96.3\%$ in cows with 2.75${\~}$3.50 BCS.

• Journal of Embryo Transfer
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• v.30 no.2
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• pp.73-82
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• 2015

This study was designed to evaluate the possibility of increase through dairy female offspring's ratio by transfer of pre-selected transferrable blastocyst that was produced by pre-selected X-bearing semen with OPU derived oocytes. Elite dairy female cow is demanded strongly compared with male, the so called, farmer wants to produce only an elite female dairy offspring as a candidate female dairy cow for producing milk. In our study, we selected 2 elite dairy bull semen from National Agricultural Cooperative Federation to pre-select X-bearing semen and 5 elite dairy female cows as donor for collecting of OPU derived oocytes. OPU derived embryo production system was carried out an aspiration of immature oocytes from 5 donor cows 2 times per week, total 200 times for 2 to 7 months by an ultrasonographic guided follicular aspiration system and then produced in vitro-produced blastocysts by in vitro maturation, fertilization and culture. Dairy donor semen selected H-319, 320 bull in National Agricultural Cooperative federation was sorted X-bearing semen by flow-cytometer and frozen for using IVF with OPU derived oocytes. Donor cows were selected 5 elite dairy cows from Gyeongju Dairy Cow Community and then disease tests such as 4 kinds of disease before selecting was checked. Oocyte proportion of grade 1 to 3 from total collected oocytes was significantly lower in donor A and B than those in donor C, D and E (82.16 and 70.03% vs. 90.0, 91.78 and 93.57%), respectively (p<0.05). However, number of oocytes per session in donor A, C and E was significantly higher than those in donor B and D ($7.77{\pm}3.26$, $5.85{\pm}2.10$ and $7.03{\pm}2.14$ vs. $4.68{\pm}2.61$ and $5.21{\pm}1.97$ oocytes), but donor A was significantly higher than donor C (p<0.05). Development to blastocyst in donor B, C and E was significantly higher than those in donor A and D (31.0, 25.0 and 25.0% vs. 14.3 and 4.5%), but donor A was not different in donor C and E (p<0.05). Nine out of 10 blastocysts (90.0%) derived from OPU blastocysts were confirmed male embryos that was induced with Y-bearing semen to confirm sex ratio only. Total 96 blastocysts derived from female bearing semen were transferred into synchronized recipients and then confirmed 42 recipients (43.8%) pregnancy rate, 36 offspring (37.5%) and 91.7% female sex ratio (33 female vs. 3 male offspring). Taken together all data, elite dairy female offspring could be produced effectively by in vitro production system between pre-selected x-bearing semen and OPU derived oocytes that would be influential breeder in the breeding of dairy farm to increase effectively elite dairy offspring ratio as well as net income in the dairy farmer.

• Microbiology and Biotechnology Letters
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• v.43 no.3
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• pp.212-218
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• 2015

Fucosyltransferases (FucTs) catalyze fucosyl transfer from guanosine-diphosphate fucose (GDP-β-L-fucose) to acceptor molecules to form fucosyloligosaccharides with α-glycosidic linkages. However, when FucT genes have been expressed in Escherichia coli, most cases have resulted in the production of inclusion bodies. In this study, to overcome this drawback, molecular chaperones were co-expressed with α1,2-fucosyltransferase (FucT2) in E. coli. For this, the pACYC184 vector, having genes for chaperones such as GroEL, GroES, DnaK, DnaJ, and GrpE, were transformed into E. coli BL21 (DE3) star harboring pHFucT2, including the FucT2 gene from Helicobacter pylori 26695. The results from SDS-PAGE showed that 5 chaperones were successfully expressed and the soluble fraction of FucT2 was also increased. HPLC analysis revealed that the coexpression of chaperone proteins resulted in a 5-fold increase in the total activity of fucosyltransferase in E. coli. In conclusion, the FucT2 expression system developed in this study can be used as a useful tool for the synthesis of fucosyloligosaccharides.

• Korean Journal of Veterinary Service
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• v.31 no.4
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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.

• Korean Journal of Breeding Science
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• v.42 no.3
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• pp.226-230
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• 2010

'Gwanghan' (Avena sativa L.), a winter oat for forage use, was developed by the breeding team at the Department of Rice and Winter Cereal Crop, NICS, RDA in 2009. It was derived from a cross between 'Early80' and 'Gwiri23'. A parent, 'Early80', has early heading and high yielding, while Gwiri23 has mid-heading with large-size grain. Subsequent generations followed by the cross were handled in bulk and pedigree selection programs at Suwon and Yeoncheon, respectively. A line, 'SO96016-B-112-YB-12-7', was selected for earliness, cold tolerance, and good agronomic characteristics and designated as a line name of 'Gwiri64'. The line was subsequently evaluated for cold tolerance, earliness, and forage yield in four different locations, Yesan, Iksan, Kimjae, and Jeju, from 2007 to 2009, and finally named as 'Gwanghan'. Over 3 years, the average forage dry matter yield of 'Gwanghan' harvested at milk-ripe stage was $16.4MT\;ha^{-1}$, compared with $15.4MT\;ha^{-1}$ of check cultivar 'Samhan'. 'Gwanghan's' heading date is about 3 days earlier than that of check cultivar, and is adapted primarily for winter cultivation in the mid-southern regions of Korean peninsula.