• Journal of Embryo Transfer
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• v.20 no.3
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• pp.303-309
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• 2005

This study was conducted to examine the effects of OA on metaphase of meiosis II and the mitochondrial activity of cytoplasm in bovine cumulus oocytes complexes(COCs) during in vitro maturation. Hanwoo COCs were collected from the slaughterhouse cow ovaries and matured in TCM199 supplemented with $0.1\%$ PVA, 0.2 uM, 2 uM, 20 uM OA for the maturation rate of OA concentration. For the maturation effects between OA and cycloheximide(CX), COCs were matured in TCM199 with 25 ug/mL CX, 25 ug/mL CX (6 hrs culture) plus 2 uM OA or 2 uM OA only at a atmosphere $5\%\;CO_2,\;95\%$ air $39^{\circ}C$ for 6, 12, 24 hrs. To evaluate the nuclear types of matured COCs, cumulus cells were removedby $0.5\%$ hyaluronidase sol. and oocytes were fixed in 1:3 acetic acid ethyl alcohol for 30 sec. and then stained with $0.1\%$ basic Fuchsin sol. For the detection of fluoriscent intensity (FI) of matures oocytes, cumulus cells were removed same as performed above and were stained with 20 nM mite tracker for 20 min. at $39^{\circ}C$. Mitochondrial activity of FI in matured oocytes was imaged by laser conforcal microscopy (Fluoview, Olympus, Japan) and were measured scanned face on 5 um from median to endpoint of oocytes. Statical analysis of nuclear types observed the three replicates was carried out with ANOVA and Fisher's protected least significant difference test using the STATVIEW program. FI of matures oocytes was compared the multiples of the least intensity among the measured oocytes. Maturing in TCM199 supplemented with $0.1\%$ PVA, 0.2 uM, 2 uM, 20 uM OA, metaphase B were showed 72.0, 50.0, 70.0, $68.8\%$, respectively and there were different significant(p<0.05). In the case of treatment with OA and CX, metaphase were $73.8\%,\;8.2\%,\;45.5\%,\;73.7\%$ in $0.1\%$ PVA-TCM199, 25 ug/mL CX, 25 ug/mL CX plus OA or 2uM OA only, respeclively. FI was revealed the increasing tendency during the process of maturation. Whereas FI in CX was decreased about 3 times compared to the other treatments of 6 hrs maturation. We conclude that OA regulates bovine COCs maturation and induces the mitochondrial activity during the process of maturation.

• Journal of Embryo Transfer
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• v.29 no.3
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• pp.265-271
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• 2014

Implementation of smart embryo technologies in cattle e.g. ovum pick-up followed by in vitro embryo production (OPU-IVP). Seasonal variation is important factor for follicular growth, oocytes quality, quantity and developmental competence. Therefore the aim of present study was carried out to investigated whether the seasons (hot and cool) effect on follicular development, oocyte recovery and subsequent embryo development. Follicular oocytes were aspirated from Korean native cows (Hanwoo) by the ovum pick-up (OPU) method, which was performed 24 times during two different seasons, the hot (July to September) and cool (October to December), from OPU donors. The recovered oocytes were classified according to morphological categories and used for in vitro embryo production (IVEP). The mean number of total follicles was significantly higher (p<0.05) during the hot season ($18.32{\pm}2.26$) compared to cool season ($15.41{\pm}3.34$). Furthermore, seasons did not significantly effect on the number of oocytes recovered (hot season: 41.16% vs. cool season: 46.14%). However, the average number of Grade A oocytes was significantly greater during hot ($1.75{\pm}1.86$) season compared to the cool season ($1.00{\pm}1.46$), but there was no significant difference of other grades oocytes. The cleavage rate (hot: 66.67% vs. cool: 63.3%) and embryo development (hot: 58.95% vs. cool: 56.97%) did not differ significantly between the seasons. In conclusion, the results of present study suggest that the season (hot and cool) does not have effects on the oocyte recovery and embryo developmental competence of in vitro cultured embryos.

• Journal of Animal Science and Technology
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• v.45 no.5
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• pp.689-694
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• 2003

Weight records of Hanwoo cows from birth to 36 months of age collected in Daekwanryeong branch, National Livestock Research Institute(NLRI) were fitted to Gompertz, von Bertalanffy and Logistic functions. For the growth curve parameters fitted on individual records using Gompertz model, the mean estimates of mature weight(A), growth ratio(b) and growth rate(k) were 383.42 ${\pm}$ 97.29kg, 2.374 ${\pm}$ 0.340 and 0.0037 ${\pm}$ 0.0012, respectively, and mean estimates of body weight, age and daily gain rate at inflection were 141.05 ${\pm}$ 35.79kg, 255.63 ${\pm}$ 109.09 day and 0.500 ${\pm}$ 0.123kg, respectively. For von BertalanfTy model, the mean estimates of A, b and k were 410.47 ${\pm}$ 117.98kg, 0.575${\pm}$0.057 and 0.003 ${\pm}$ 0.001, and mean estimates of body weight, age and daily gain at inflection were 121.62 ${\pm}$ 34.94kg, 211.02 ${\pm}$ 105.53 and 0.504 ${\pm}$ O.l24kg. For Logistic model, the mean estimates of A, b and k were 347.64 ${\pm}$ 97.29kg, 6.73 ${\pm}$ 0.34 and 0.006 ${\pm}$ 0.0018, and mean estimates of body weight, age and daily gain at inflection were 173.82 ${\pm}$ 37.25kg, 324.47 ${\pm}$ 126.85 and 0.508 ${\pm}$ 0.131kg. Coefficients of variation for the A, b and k parameter estimates were 25.3%, 14.3% and 32.4%, respectively, for Gompertz model, 28.70/0, 9.9% and 33.3% for von Bertalanffy model, and 27.9°/0, 5.0% and 30.0% for Logistic model.

• Korean Journal of Organic Agriculture
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• v.21 no.4
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• pp.629-646
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• 2013

This study was conducted to investigate effects of terpenoids-rich plant extracts (TRPE) on the in vitro ruminal fermentation characteristics and methane production. The ruminal fluid was collected from a cannulated Hanwoo cow fed concentrate and timothy in the ratio of 6 to 4. The TRPE as Mint (Mentha arvensis var. piperascens), Pine (Pinus densiflora), Japan cedar (Cryptomeria japonica), Sichuan pepper (Zanthoxylum piperitum), Hinoki cypress (Chamaecyparis obtuse) and Japanese black pine (Pinus thunbergii) were used in this study. The 15 mL of mixture, contains McDougall buffer and rumen fluid in the ratio of 2 to 1. The mixture was dispensed anaerobically 50 mL serum bottles and it is contained 0.3 g timothy substrate and 5% TRPE. The bottles were incubated at $39^{\circ}C$ for 3, 6, 9, 12, 24, 48 and 72 hours. The pH value decrease by increased incubation times and the pH values at all times were significantly (p<0.05) higher in treatments than in control. The digestibility of dry matter at 3 hours was significantly (p<0.05) higher in mint treatment than in control. Productions of total gas and carbon dioxide at before 12 hours was significantly lower (p<0.05) in treatments than in control. The methane production at 24 hours was significantly (p<0.05) lower in treatments than in control. The concentrations of acetic acid and propionic acid at 24 hours were significantly higher (p<0.05) in mint and pine treatments than in control. In conclusion, the terpenoid-rich plant extracts were shown to decreased methane emission and without adversely affected ruminal fermentation. Therefore, the terpenoid-rich plant extracts as mint and pine were shown to decreased methane production and it has potential possibility for ruminal fermentations.

• Korean Journal of Organic Agriculture
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• v.25 no.4
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• pp.901-916
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• 2017

This study was conducted to evaluate natural plant extracts for methane gas reduction in ruminants. Rumen fluid was collected from cannulated Hanwoo cow ($450{\pm}30kg$) consuming 400 g/kg concentrate and 600 g/kg timothy. The 15 ml of mixture comparing McDougall's buffer and rumen fluid in the ratio 2 to 1, was dispensed anaerobically into 50 ml serum bottles. Rumen fluid contents were collected and in vitro fermentation prepared control (timothy, 300 mg), ginseng, balloon flower, yucca plant, camellia, tea plant and ogapi extracts were added at the level of 5% against 300 mg of timothy as a substrate (v/w) and incubated for 3, 6, 9, 12, 24, 48, and 72 h. In vitro pH values range 6.55~7.41, this range include rumen titration. The dry matter digestibility was not differ between all treatments and control. Total gas emission was significantly higher (p<0.05) in ginseng and balloon flower treatments on 24 h than in control. Carbon dioxide emission was not differ all treatments on 9 h than in control and significantly higher (p<0.05) yucca plant, camellia and tea plant treatments on 12 h than control. Methane emission was not differ all treatments on 6 h than in control. The rumen microbial growth rate was significantly higher (p<0.05) in ginseng, balloon flower on 12 h and significantly higher (p<0.05) in ginseng, yucca plant, tea plant and ogapi treatments on 24 h than in control. Total VFA was significantly higher (p<0.05) in tea plant and ogapi treatments on 12 h than in control and significantly higher (p<0.05) in ginseng, balloon flower treatments on 48 h than in control. Acetic acid was significantly lower (p<0.05) in ginseng and balloon flower treatments on 24 h than in control. Propionic acid was significantly higher (p<0.05) in ginseng and balloon flower treatments on 48 h than in control. As a results, sixth natural plant extracts had no significant effect dry matter digestibility and negative on rumen fermentation, but not effect methane reduction.

• Journal of Animal Environmental Science
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• v.18 no.1
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• pp.45-56
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• 2012

This study was conducted to give basic information of the animal manure management by manure units determination for recycling farming in Cheorwon-gun. Manure units (MU) are used in the permitting, registration, and the environmental process because they allow equal standards for all animals based on manure nutrient production. An MU is calculated by multiplying the number of animals by manure unit factor for the specific type of animal. The manure unit factor for MU determination was determined by dividing amounts of manure N produced 80 kg N/year. Conversion to manure units is a procedure used to determine nutrient pollution equivalents among the different animal types. In this study, the manure unit factor based on nitrogen in Hanwoo, dairy cow, pig were 0.36, 0.8 0.105, respectively. The analysis of manure unit per ha shows that the N loading by MU is quite different by region. The nitrogen loading of manure unit (MU) per ha of cultivated land was the highest in the Galmal-eup on province with 2.4 MU/ha, which is higher than the appropriate level. The Seo-myeon province came next with 1.92 MU/ha. To be utilized as a valid program to build the recycling farming system, diverse measures shall be mapped out to properly determine manure units, evaluate N-loading and to properly manage their nutrient balance of each region.

• Journal of Embryo Transfer
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• v.20 no.2
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• pp.169-176
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• 2005

Loop-mediated isothermal amplification (LAMP) is novel DNA amplification methods that amplifies a target sequence specifically under isothemal condition. The present study was to assess the in vitro viability afier biopsy and sexing rate of different types of embryo biopsied. In vivo compact morulae and blastocyst embryos were obtained from Korean Native Cow (KNC) superovulated with FSH (Antorin, R-10) on 7 Day after artificial insemination. in vitro compact morulae and blastocyst embryos were obtained with KNC or Holsteins that were gained on 6, 7 or 8 day after in vitro fertilization(IVF) with frozen semen. Biopsy of bovine embryo was carried out in a $80{\mu}l$ drop with $Ca^{2+}-Mg^{2+}$ free D-PBS and the viability of biopsied embryos were evaluated in IVMD (IFP, Japan) medium at 12 hrs culture time. The sex ratio of biopsied Hanwoo embryos were male vs. female of $43.5\%\;vs.\;56.5\%$ in vivo and $33.9\%\;vs.\;49.2\%$ in vitro respectively, and male rate of biopsied Holstein embryos were significantly higher than female $(70.8\;vs.\;29.2\%)$. and indefinite rate of in vitro embryos was $16.9\%$ and in vivo was not. The degeneration rate of biopsied embryo, in vitro embryos were significantly higher than in vivo $(13.2\%\;vs,\;0.0\%,\;p<0.05)$. The survivability of in vivo embryo were between biopsied following punching method was significantly (P<0.05) higher than bisection method produced embryos $(100\%\;vs.\;83.3\%)$ and in vitro had no difference. However, the degeneration rate of biopsied embryo by bisection method was significantly higher than punching methods between in vivo and in vitro $(16.7\;vs.\;22.6\%,\;respectively,\;p<0.05)$. In conclusion, these results indicate that punching method was optimal and survivability after embryo biopsy was useful for reducing the damage caused by the embryo biopsy procedure for LAMP-based embryo sexing.

• Journal of agriculture & life science
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• v.50 no.2
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• pp.95-105
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• 2016

This study was conducted to evaluate the effects of nitrate-rich plants extracts on the in vitro rumen fermentation characteristics and rumen methane production. The extracts of nitrate-rich plants, as potato, carrot, chinese cabbage, lettuce and spinach were used in this study. The ruminal fluid was collected from a cannulated Hanwoo cow fed concentrate and timothy in the ratio of 6 to 4. The 20mL of mixture, comparing McDougall's buffer and rumen fluid in the ratio 2 to 1, was dispensed anaerobically 50mL serum bottles containing 0.3g of timothy substrate and extracts of nitrogen-rich plants. The serum bottles were incubated 39℃ for 9, 12, 24, 48 hours. The pH value was decreased by increased incubation times and normal range to 6.31 to 6.96. The dry matter digestibility was significantly(p<0.05) lower in chinese cabbage than in control at 9h incubation time. Ammonia concentration was significantly(p<0.05) lower in potato, chinese cabbage, lettuce than in control and the rumen microbial growth rate was significantly(p<0.05) higher in carrot than in control at 24h incubation time. The concentrations of acetate and propionate was significantly(p<0.05) lower in treatment than in control. The concentration of butyrate was showed a different pattern depending on treatments. Total gas emissions was significantly(p<0.05) lower in chinese cabbage, lettuce, spinach than in control at 12h, 24h incubation time. Methane production was significantly(p<0.05) lower in potato, chinese cabbage, spinach than in control, carbon dioxide production was significantly(p<0.05) lower in treatment than in control. In conclusion, supplementation of the nitrate-rich plant extracts in ruminal fermentation in vitro resulted in decreasing the methane production without adversely affecting the fermentation characteristics. Particularly the chinese cabbage extract was regard as a potential candidate for reducing the methane emission in ruminants.