• Korean Journal of Soil Science and Fertilizer
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• v.19 no.1
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• pp.56-62
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• 1986

Fertilization response on production and quality of flue-cured tobacco as to different level of nitrogen, phosphorus, and potassium were studied in a field experiment. The results were summarized as follows: 1. Growth and yield of flue-cured tobacco were significantly better in higher nitrogen fertilization levels, regardless of soil fertility, but the negative correlation was recognized between the quality of leaves and the amounts of nitrogen application. While, both fertilizers of phosphorous and potassium did not have should little effect on the tobacco yield and quality. 2. The optimum ratio of N, P, and K fertilizer applications were decided by the appearance of the proper yield and the best quality of tobacco leaves. The proportion of N:P:K was 2:1:4. 3. The single effect (Complete plot minus Non-fertilized plot) of N, P, and K on yield and quality of cured leaf was greatly affected by nitrogen, but the combined effect (Nutrient deficiency plot minus Non-fertilzier plot) of that were only slightly affected by P and K.

• Journal of Embryo Transfer
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• v.27 no.1
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• pp.29-35
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• 2012

The purpose of this study was to examine the effect of growing stages of the Korean Native Striped Bull (KNSB) on the freezability and fertility of frozen-thawed semen. First, we investigated the total motility (TM) and progressive motility (PM) according to the diluent used for semen freezing. Second, we examined the effect of the age of KNSB on semen volume, TM and PM of fresh and frozen-thawed semen. Third, we examined the effect of frozen semen from the different age of KNSB on the $in-vitro$ fertilization rate, and the artificial insemination pregnancy rate. The diluents used in this experiment were Triladyl$^{(R)}$ and Tris-egg yolk extender (EYE). Semen was collected from 5 KNSB in the growing stage (15 months) and 5 adult KNSB (36 months). When Triladyl or Tris-EYE extender was used for semen freezing, there was no difference of the mean TM and the mean PM. However, the mean TM was significantly higher in Bull No. 1885 than Bull No. 4283 ($p$ <0.05). The mean volume of semen collected from the 15-month-old bulls (2.3 ml) was significantly lower ($p$ <0.05) than that from the 36-month-old bulls (5.0 ml). The mean semen concentration was similar for the 15-month-old ($2.1{\times}10^9$ spermatozoa/ml) and 36-month-old ($1.8{\times}10^9$ spermatozoa/ml) bulls. For the 15-month-old and 36-month-old bulls, the mean TM of fresh semen were 93.7% and 88.3%, respectively, and the mean PM were 97.0% and 88.3%, respectively; the 15-month-old bulls showed a particularly high PM ($p$ <0.05). For the 15-month-old and 36-month-old bulls, the mean TM (56.0% and 58.0%, respectively) and the mean PM (64.0% and 70.7%, respectively) of frozen-thawed semen did not differ. The development rates of embryos after $in-vitro$ fertilization and the pregnancy rate after artificial insemination using frozen-thawed semen did not differ according to the bull's age. In summary, semen volume differed according to the bull's age, but semen concentration and survival rate, the $in-vitro$ fertilization rate, and the pregnancy rate did not differ according to the stripe bull's age. Accordingly, semen from bulls in the growing stage can be collected and frozen for the preservation and multiplication of rare livestock.

• Clinical and Experimental Reproductive Medicine
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• v.27 no.3
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• pp.261-265
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• 2000

Objective: This study was performed to evaluate the influence of maternal age on embryo quality and the frequency of multiple pregnancy in IVF-ET program. Method: 86 conventional IVF-ET cycles were divided into three groups according to the age by 5 year (group A: 26-30, group B: 31-35, group C: 36-40 yrs). The in vitro fertilization and development outcome (fertilization, cleavage and high quality embryo rate) and the pregnancy outcome (pregnancy, implantation, G-sac/high quality embryo and multiple pregnancy rate) were examined. And then, these results were compared among the groups. Results: The rates of fertilization (62.7, 68.5 and 65.4%, respectively) and cleavage (95.6, 97.6 and 98.0%, respectively) were not different among the groups. And the high quality embryo (HQE) rate also was not different among the groups (61.8, 62.9 and 62.8%, respectively). The pregnancy rate of group C (23.3%) was significantly lower than that of group A (41.2%) and B (48.7%). And the implantation rate was significantly decreased with advance in maternal age (group A; 17.3%, B; 12.6% and C; 6.0%). The G-sac/high quality embryo rate was significantly higher in group A (70.8%) when compared to group B (32.2%) and C (40.0%). On the other hand, the multiple pregnancy rate was significantly lower in group C (14.3%) when compared to group A (71.4%) and B (36.8%). Conclusion: The pregnancy rate was significantly decreased over 35 years. The G-sac/HQE and multiple pregnancy rate were significantly high below 31 years. Thus, these results suggest that the number of high quality embryo transferred should be limited by the age and another criteria for embryo quality evaluation were required for single embryo transfer.

• Korean Journal of Animal Reproduction
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• v.26 no.2
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• pp.173-182
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• 2002

The mechanisms underlying of the visual assessment and resulting in optimum embryonic development following in vitro maturation, fertilization, and culture are unclear, It was known that in vitro produced embryos show more frequent occurrence of fragmentation, which resulted in poor developmental potential and decreased implantation rate. The objective of this study was to investigate the apoptotic rates in bovine blastocyst derived from in vitro fertilization (IVF) and nuclear transfer (NT). In addition, the expression levels of Bcl-2 and Bax gene were investigated in the blastocyst to confirm their potential roles in the regulation of apoptosis during preimplantation embryonic development. Analysis of apoptosis was carried out by using terminal deoxynucleotidyl transferase mediate dUTP nick end labeling (TUNEL) method. The levels of Bcl-2 and Bax gene in the blastocyst derived from IVF and NT were determined by RT-PCR. The proportion of TUNEL positive signal in blastocyst derived from NT was significantly higher than that in blastocyst derived from IVF (p<0.001). Bcl-2 expression level of blastocyst derived from IVF was higher than that of blstocyst derived from NT. However, high expression level of Bax was observed in the blastocyst derived from NT. These results indicates that apoptosis is more responsible for fiagmentation in bovine blastocyst derived from NT than IVF. These results suggested that the increase of developmental failure followed by NT could be caused by nuclear fragmentation as apoptosis.

• Korean Journal of Soil Science and Fertilizer
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• v.48 no.6
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• pp.641-647
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• 2015

Monitoring of soil fertility by long-term application of fertilizers is necessary to improve the fertility of soil and the productivity of crop. The objective of this study was conducted to investigate the changes of exchangeable Mg by continuous application of fertilizers from 1969 to 2014. The treatments were no fertilization (No fert.) and fertilization (NPK, NPK+C, NPK+S, and NPK+CS). The concentration of exchangeable Mg in No fert., NPK+C, and NPK+S treatments tended to increase from 1965 to 1975, but decrease gradually from 1976 to 1987, and increase again after 1988. Based on these, the changes of exchangeable Mg were divided into period I ('69 ~'75), period II ('76~'87), and period III ('88~'14). Especially, exchangeable Mg decreased in the period II. This was presumed that a significant amount of Mg from topsoil were leached into subsoil by break of plow pan and some of subsoil was incorporated into topsoil according to change of plowing depth by replacement of tillage machinery. It could be possible that exchangeable Mg in NPK, NPK+S, and NPK+CS was accumulated in the depth of 15~20 cm. For the period III, exchangeable Mg in No fert., NPK, NPK+C, NPK+S, and NPK+CS treatments increased at rates of 0.013, 0.018, 0.015, 0.023, and $0.024cmolckg^{-1}{\cdot}yr^{-1}$ respectively. Exchangeable Mg level in NPK+S was lower than the other treatments in the period I and period II, but higher than in the period III. This result was attributed to replacement of silicate fertilizer type from wollastonite (Mg 0.3%) to silicate fertilizer (Mg 3%). Also, exchangeable Mg level of No fert. treatment increased, which showed that Mg concentration of irrigated water had the greatest impact on Mg accumulation of soil. Recently, Mg level of irrigated water tended to increase, indicating that Mg concentration of water will affect greatly the concentration of exchangeable Mg of soil in the future. Like these, the changes of exchangeable Mg were greatly influenced by agricultural environment such as plowing depth, plow pan, content of fertilizer, and quality of irrigated water. Considering these agricultural environment, the proper management of soil is needed for the improvement of soil fertility and crop productivity.

• Journal of Marine Life Science
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• v.6 no.2
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• pp.80-87
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• 2021

In this study, the toxic effects of PFOA and PFOS potassium salt on Mesocentrotus nudus using 10 min-fertilization rate and 48 h-normal embryogenesis were confirmed through the calculation of toxicity values such as Non-observed effective concentration, Low-observed effective concentration, and 50% of effective concentration. The case of 10 min-fertilization rate and 48 h-normal embryogenesis showed the concentration-dependent reduction pattern when exposed to PFOA and PFOS potassium salt, in tested concentration, respectively. The EC50 values of 10 min-fertilization rates for PFOA and PFOS potassium salt were 1346.43 mg/l and 536.18 mg/l, respectively, and the EC50 values of 48 h-normal embryogenesis were 42.67 mg/l and 17.81 mg/l, respectively. Both toxicity test methods showed high toxicity sensitivity to PFOS potassium salt. Recent studies have shown that the concentration of PFOA and PFOS in the marine environment has continuously decreased, and it is not enough to show acute toxicity to sea urchin. However, PFOA and PFOS have a very long half-life and can accumulate throughout the life of marine life, so it is still observed at a high concentration in shellfish. Therefore, a study on chronic toxicity through the whole-life cycle of marine organisms in coastal environments should be needed.

• Korean Journal of Environmental Agriculture
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• v.26 no.4
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• pp.306-312
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• 2007

To investigate the effect of N fertigation on the growth, yield, and water and nitrogen use efficiencies during tomato cultivation, seedlings were transplanted in a sandy loam soil under plastic film house condition. 0, 88, 132, 176, $220\;kg\;ha^{-1}$ N rates, which correspond to 0 (NF0), 40 (NF40), 60 (NF60), 80 (NF80), 100% (NF100) N level of soil test-based N fertilization, were injected weekly through drip irrigation system for 15 weeks in N fertigation system, and the control (conventional N treatment) was installed for comparison. Herein, nitrogen was applied by top-dressing with 60% as a basal and 40% as additional fertilizer. There was little different in stem diameter growth among N fertigation treatments, but plant height and dry matter increased with increasing N fertigation rates as well as in N conventional treatment. Tomato yield was increased with increasing the number of marketable fruits in N fertigation treatments, and the fruit yield was maximized in NF 80 treatment ($176\;kg\;ha^{-1}$ N supply or $96.6\;mg\;L^{-1}$ N injection). Dry matter productivity and nitrogen uptake amount were significantly increased with increasing N fertigation rates. The ratio of fruits to the dry weight of whole plant was decreased with increasing N fertigation rates, but this ratio was $2.6{\sim}5.3%$ higher in N fertigation treatments than in the control. In addition, the ratios of nitrogen distributed toward fruits in N fertigation treatments were $3.7{\sim}21.7%$ higher than that of control. The apparent N recovery percentages showed significantly higher values as $71.8{\sim}102.3%$ in N fertigation treatments, compared to 45% in N conventional treatment. Water use efficiency was significantly increased by fertigation system with the maximum $361\;kg/ha\;cm^{-1}$ in NF 80, which is comparable to $324\;kg/ha\;cm^{-1}$ of the conventional treatment. Conclusively, N fertigation system was effective on increasing tomato productivity and nutrient efficiency as well as 20% reduction of N fertilization level.

• Journal of Embryo Transfer
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• v.29 no.1
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• pp.13-19
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• 2014

This study was carried out to study the survival rate of thawed Hanwoo embryos frozen by the slow-rate freezing or the cryotop vitrification method. Hanwoo cumulus-oocyte complexes were recovered from ovaries at a slaughter house, matured for 20~22 hours, fertilized with Hanwoo semen for 5~6 hours, and cultured for 7~9 days in $38.5^{\circ}C$, 5% $CO_2$ incubator. For freezing, Day 7~9 blastocysts were collected. Embryos for the slow-rate freezing were equilibrated in 1.8 M ethylene glycol (EG) with Dulbecco's phosphate-buffered saline (D-PBS). Programmable cell freezer was precooled down to $-7^{\circ}C$, and the straw was seeded during 8 minutes-holding time, and was cooled to $-35^{\circ}C$ at the cooling rate of $0.3^{\circ}C/min$, and then was plunged and stored in liquid nitrogen. Embryos for the cryotop vitrification were treated in TCM199 with 0.5 M sucrose, 16% EG, 16% dimethylsulfoxide (DMSO). Embryos were then loaded individually onto cryotop and plunged directly into liquid nitrogen. The survival rates of embryos frozen by these two freezing methods were evaluated at 12 to 24h post-thawing. The survival rates of frozen/thawed Hanwoo embryos by the cryotop vitrification method ($56.86{\pm}26.53%$) were slightly higher than those by the slow-rate freezing method ($55.07{\pm}26.43%$) with no significant difference. Using the cryotop vitrification and the slow-rate freezing of Hanwoo blastocysts on Day 7 following in-vitro fertilization (IVF) treatment, the survival rates of frozen/thawed Hanwoo embryos were $72.65{\pm}18.3%$ and $79.06{\pm}17.8%$, respectively. The survival rates by the cryotop vitrification were higher than those by the slow-rate freezing on both Day 8 and 9 with significantly higher survival rate on Day 9 (p<0.05). Using the cryotop vitrification and the slow-rate freezing of Hanwoo embryos to compare between three different blastocyst stages, the survival rates of the blastocyst stage embryos were $66.22{\pm}18.8%$ and $45.76{\pm}12.8%$, respectively with higher survival rate by the vitrification method (p<0.05). And the survival rate of expanded blastocysts was higher than those of early blastocysts and blastocysts in two freezing methods with significantly higher survival rate by the slow-rate freezing method (p<0.05).

• Korean Journal of Animal Reproduction
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• v.11 no.1
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• pp.73-84
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• 1987

This experiment was carried out to investigate the effects of hormone addition(FSH, HCG, estrogen and progesterone) and composition (BSA and FCS) of mKRB on the in vitro maturation and fertilizability of follicular oocytes of the Korean native cattle. The ovaries were removed at a slaughterhouse, returned to laboratory in a thermostat (30-35$^{\circ}C$) within 4 hr, and collected by aspirating normal follicles which had diameters of 1 to 6 mm. The oocytes with cumulus cells were cultured for 8, 16, 24 and 30 hr in a modified KRB solution containing BSA or FCS and hormones. The in vitro matured oocytes in mKRB containing FCS, FSH and steroids were transferred in the rabbit uterus for examination of their in vivo fertilizability with bovine sperm preincubated 4 to 6 hr in the rabbit uterus. 1. The mean number of oocytes collected per cattle was 6.5 from 1-3mm follicles, 1.3 from 4-6mm follicles, and total was 7.7. 2. The meiotic division at 16hr-cuture in the oocytes from 1-3mm follicles was slightly stimulated by the addition of FSH in mKRB + BSA solution compared with the control. At 30hr-culture, their maturation rates(%Met II) were also increased by FSH of 1 $\mu\textrm{g}$/ml(38.4%) and 5$\mu\textrm{g}$/ml(35.7%) as compared with the control (21.4%). The maturation rate at 30hr-culture in the oocytes from 4-6mm follicles was 53.8% and 57.1% by the FSH addition of 1$\mu\textrm{g}$/ml and 5$\mu\textrm{g}$/ml, respectively. These rates were similar with the control(57.1%), but higher than those of oocytes from 1-3mm follicles. 3. The meiotic division at 16hr-culture in the oocytes from 1-3mm follicles was stimulated by the HCG addition of 1IU/ml and 5IU/ml. However, the maturation rate at 30hr-culture was greatly decreased by the HCG addtion (26.6% and 13.3%) compared with the control(53.3%) and these rates (30.8%) in the oocytes from 4-6mm follicles were also lower than that fo the control(58.3%). 4. Low maturation rate (37.5%) of the oocytes cultured in mKRB containing BSA and 5IU/ml HCG was increased (55.0%) when 15% FCS with HCG was added to mKRB instead of BSA. 5. When 16hr-cultured oocytes in mKRB containing BSA and gonadotropins (5$\mu\textrm{g}$/ml FSH and 5IU/ml HCG) were transferred in the medium without gonadotropins and recultured for 16hr, the maturation rate of HCG-treated oocytes was greatly improved. 6. The maturation rates of oocytes were greatly affected by steroids. The combined addition of FCS+FSH+estrogen or +progesterone to mKRB increased the maturation rate compared with the combination of BSA+FSH or FCS+FSH in mKRB. 7. The fertilization rate, presence of pronuclei, was increased by the combination of FCS+FSH+p in mKRB as compared with that (5.6%) of BSA+FSH and the rates of FCS+FSH+steroids ranged from 12.5 to 17.6%.

• Journal of Embryo Transfer
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• v.16 no.3
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• pp.163-172
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• 2001

The present study was carried out to examine the effects of $O_2$concentrations and culture media on in vitro maturation and embryo development of porcine follicular oocytes. The results were summarized as fellows : 1. The rates of GVBD and nuclear maturation in NCSU-23 and TCM-199 media with 10% PFF under the conditions of 5% and/or 20% $O_2$concentrations were not different among the each treatment groups(P>0.05). 2. The rates of polyspermy and mean numbers of penetrated sperm were significantly lower in NCSU-23 medium than in TCM-199 medium (P>0.05). However, the rates of polyspermy and mean numbers of penetrated sperm were not different between 5% and 20% $O_2$concentrations. 3. The rates of blastocyst formation at day 7 after in vitro fertilization were significantly higher in NCSU-23 medium under the condition of 20% $O_2$concentration than in TCM-l99 medium under the condition of 5% or 20% $O_2$concentrations. However, the rates of blastocyst formation and total cell numbers in blastocysts were not different between 5% and 20% $O_2$concentrations. In conclusion, the use of NCSU-23 medium under the condition of 20% $O_2$concentration was beneficial for porcine oocyte maturation and in vitro development.