• Clinical and Experimental Reproductive Medicine
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• v.24 no.2
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• pp.153-165
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• 1997

In male reproducible health, fertility and IVF (in-vitro fertilization), semen analysis has been most important. Semen analysis can be divided into concentration, motional and morphological analysis of sperm. The existing method which was developed earlier to analyze semen concentrated on the sperm motility analysis. To provide more useful and precise solutions for clinical problems such as infertility, semen analysis must include sperm morphological analysis. But the traditional tools for semen analysis are subjective, imprecise, inaccurate, difficult to standardize, and difficult to reproduce. Therefore, with the help of development of microcomputers and image processing techniques, we developed a new sperm morphology analyzer to overcome these problems. In this study the agreement on percent normal morphology was studied between different observers and a computerized sperm morphology analyzer on a slide-by-slide basis using strict criteria. Slides from 30 different patients from the SNUH andrology laboratory were selected randomly. Microscopic fields and sperm cells were chosen randomly and percent normal morphology was recorded. The ability of sperm morphology analyzer to repeat the same reading for normal and abnormal cells was studied. The results showed that there was no significant bias between two experienced observers. The limits of agreement were 4.1%${\sim}$-3.8%. The Pearson correlation coefficient between readers was 0.79. Between the manual and sperm morphology analyzer, the same findings were reported. In this experiments the slides were stained by two different methods, PAP and Diff-Quik staining methods. The limits of agreement were 7.2%${\sim}$-5.7% and 6.0%${\sim}$-6.3%, respectively. The Pearson correlation coefficients ware 0.76 and 0.91, respectively. The limits of agreement was tighter below 20% normal forms. In the experiments of repeatability, 52 cells stained by PAP and Diff-Quik staining methods were analyzed three times in succession. Estimating pairwise agreement, the kappa statistic for the pairs were 0.76, 0.81, 0.86, and 0.75, 0.88, 0.88 respectively. In this study it was shown that there was good agreement between manual and computerized assessment of normal and abnormal cells. The repeatability and agreement per slide of computerized sperm morphology analyzer was excellent. The computer's ability to classify normal morphology per slide is promising. Based on results obtained, this system can be of clinical value both in andrology laboratories and IVF units.

• Asian-Australasian Journal of Animal Sciences
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• v.32 no.7
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• pp.956-965
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• 2019

Objective: The main goal of this study was to provide a morphological indicator that could be used to select high-quality oocytes of appropriate meiotic and developmental capabilities in pig. The higher quality of immature oocytes, the higher success rates of in vitro maturation (IVM) and in vitro fertilization (IVF). Thus, prior to the IVM culture, it is important to characterize oocytes morphologically and biochemically in order to assess their quality. Two of the largest indicators of oocyte quality are the presence of cumulus cells and status of chromatin. To investigate the effects of porcine oocyte chromatin configurations on the developmental capacity of blastocysts, we assessed oocyte chromatin status according to follicle size and measured the developmental potency of blastocysts. Methods: To sort by follicle size, we divided the oocytes into three groups (less than 1 mm, 1 to 3 mm, and more than 3 mm in diameter). To assess chromatin configuration, the oocytes were assessed for their stages (surrounded nucleolus [SN] germinal vesicle [GV], non-surrounded nucleolus [NSN] GV, GV breakdown, metaphase I [MI], pro-metaphase II [proMII], and metaphase II [MII]) at different maturation times (22, 44, and 66 h). To assess the development rate, oocytes of each follicle size were subjected to parthenogenetic activation for further development. Finally, GV oocytes were grouped by their chromatin configuration (SN, SN/NSN, and NSN) and their global transcriptional levels were measured. Results: SN GV oocytes were more suitable for IVF than NSN GV oocytes. Moreover, oocytes collected from the larger follicles had a greater distribution of SN GV oocytes and a higher developmental capacity during IVM, reaching MII more quickly and developing more often to blastocysts. Conclusion: Porcine oocytes with high-level meiotic and developmental capacity were identified by analyzing the relationship between follicle size and chromatin configuration. The porcine oocytes from large follicles had a significantly higher SN status in which the transcription level was low and could be better in the degree of meiotic progression and developmental capacity.

• Korean Journal of Veterinary Service
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• v.44 no.4
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• pp.227-237
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• 2021

Artificial insemination of Korean native cattle (KNC) is the predominant method for breed improvement. However, industrialization of embryo production and transfer is necessary to utilize the genetic potential of KNC. The aim of this study was to examine associations between KNC donor cows and ovum pick-up (OPU) conditions, in-vivo oocyte recovery, and embryo development. Oocyte recovery and blastocyst development rates were higher at 50 and 60 mmHg OPU vacuum pressure than at 40 mmHg, which was, however, not significant. Regarding follicle growth, injection of 500 ㎍ GnRH 36 hours before OPU significantly increased the number of OPU oocytes from an average of 4.6 to 7.6 (P<0.05); no significant difference in embryo development rates was observed. Significant differences were observed in the numbers of OPU oocytes, embryo development rates, and transplantable blastocysts per individual among nine KNC donors (P<0.05). Furthermore, although there was no difference in OPU oocyte recovery intervals in approximately 2~8 weeks, the number of recovered oocytes significantly decreased at the 12-week interval (P<0.05); there was no difference in embryo development rates. The number of oocytes and embryonic development rates only tended to decrease until the seventh OPU session, but decreased significantly until the eighth session (P<0.05). The average pregnancy rate after transfer of OPU-derived in-vitro embryos into recipient cows was 41.8%. To improve the efficiency of OPU egg recovery and in-vitro embryo production, considering KNC donor characteristics, vacuum pressure of 60 mmHg, GnRH pretreatment to induce follicle growth, and effective OPU egg recovery up to seven times at intervals of 2~4 weeks appears to be most suitable. This study may facilitate the industrialization of KNC embryo production and transfer using high-quality cows.

• Journal of the Korea Organic Resources Recycling Association
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• v.30 no.1
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• pp.5-11
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• 2022

This research evaluated the long-term application effects of different soil amendments on yield, dissolved organic carbon, nitrogen and soil organic carbon stock in rice paddy. The experiment consisted of four different fertilizations; Inorganic fertilization (NPK), NPK+Lime (NPKL), NPK+Silicate (NPKS), NPK+Compost (NPKC). There was no significant difference in rice yield between the treatment groups in 1995, but the rice yields in the NPKL and NPKC treatments in 2019 increased by 4.3% and 14.3% compared to NPK. In terms of soil properties, the pH of NPKS(6.7) and NPKL(6.4) in 2019 increased the most compared to the soil pH before experiment(5.2). The organic matter(OM) content from NPKC treatment increased upto 34 and 27 g kg^-1 in year of 1995 and 2019, respectively, compared to before the test. In NPKS and NPKL treatment, labile carbon and nitrogen content, used as a soil quality indicator, increased by 1.1-1.9 times over the control. From these result, it is suggested that type and application rate of soil amendment should be determined based on the soil analysis before cultivation for sustainable agricultural environment and productivity.

• Journal of Practical Agriculture & Fisheries Research
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• v.20 no.2
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• pp.105-114
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• 2018

The aim of this study was to evaluate the change of phenolic compounds after phosphite treatment on Cheorwon onion. Onion is a perennial plant belonging to the lily family. It is native to Persia of Southwest Asia. It is widely cultivated in the temperate regions of the world. Onion is a good name for the 'Okchong' to drop blood cholesterol and cardiovascular blood flow to increase the prevention of adult diseases. Cheorwon area is inland, but it has high continental climate due to its high altitude. Therefore it is said that the onion cultivated in this region has higher sugar content and higher taste than onion grown in the southern region. Phosphorus components are particularly important ingredients for promoting muscle development. However, if the phosphoric acid content of the soil part is maintained to a large extent until the harvest, the competition of the nutrients tends to cause decay of the root part. Therefore, it is important to improve the quality and shelf life of onion by inducing nutrient balance by applying foliar fertilization method on the reducing phosphorus at harvest time. In this study, acidity was controlled by diluting phosphorous acid(H₃PO₃) and potassium hydroxide(KOH), followed by leaf surface treatment with phosphite on onion. In this study, the concentration of phosphite was diluted to 500, 1,000, 1,500ppm and sprayed three times over the onion leaves in May 2018 using an atomizer and harvested at the end of June, and the phenolic compounds were analyzed by HPLC. As a result, the content of quercetin, one of the important substances in onion, was phosphite 500ppm(179.70㎍/g), 1,000(150.27), 1,500(105.95). The contents of caffeic acid, p-coumaric acid, ferulic acid, rutin, kaempferol, and sugar content were higher in the treatments than in the control. Therefore, the phosphite does not have a great influence on the growth, but it may play a role as a method of achieving balance with nitrogen in the rainy season by supplying the role of the material catalyst and the water soluble phosphoric acid and the potassium in the influence of the material change.

• Korean Journal of Ichthyology
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• v.36 no.3
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• pp.209-219
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• 2024

A survey was conducted in 2011 to investigate the reproductive ecology and spawning behavior of the natural monumental and endangered species, Cobitis choii. C. choii exhibits sexual dimorphism with the development of lamina circularis on the second pectoral fin ray in males at one year old, facilitating easy differentiation between males and females. A total of 1,288 females and 881 males were collected from January to December, resulting in a sex ratio of 1 : 0.68. The spawning season was estimated to be from late June to early July (spawning peak from late June, with water temperatures ranging from 22 to 25℃), coinciding with a sharp decrease in the condition index. Gonadal maturity was observed with maturity indices of 17.5±2.11% for females and 2.9±0.46% for males, with fecundity of 2,444±838 eggs and egg size of 0.75±0.05 mm. Spawning behavior was induced by hormone injection (Human Chorionic Gonadotropin, HCG) in laboratory conditions. Experimental results revealed that males and females paired in a 1 : 1 ratio before spawning. Males pressed the abdomen of females using their lamina circularis to induce spawning, followed by immediate fertilization. Spawning behavior was repeated 15 to 25 (average 20) times per pair of males and females, and 80 to 120 eggs were spawned per time, and the total amount of spawned eggs were 2,500±250 on average.

• Asian Journal of Turfgrass Science
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• v.22 no.1
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• pp.65-74
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• 2008

Traffic management is becoming an important issue in turfgrass practise on golf course. The objective of this study was to investigate the combined impact of different traffic times and types of fertilization. Traffic treatment was applied in morning (AM), noon, and afternoon (PM). Fertilizers used include faster-release fertilizer (21-17-17) and slow-release fertilizers (12-6-18, 11-3-22, 20-3-20, 10-3-10). Experiment was conducted from Oct. 1 to Nov. 30, 2005 on a nursery putting green of Incheon Grand Golf Club. The growth and quality of creeping bentgrass (Agrostis polustris cv. 'Seaside II') were evaluated on visual leaf color, leaf texture, shoot density, and root length. The measurement at 20 days after treatment, turfgrass color and leaf texture showed the best result on 10-15-10 + Noon-traffic plot. Turf quality and traffic tolerance were not different at Am and Pm traffic treatment. However, traffic stress in early morning and late evening caused the most severe damage to the turfgrass. Shoot density was the highest in 10-15-10 + Noon-traffic treatment but root length was not different among treatments after 30 day measurement. Among the fertilizers, slow release fertilizer resulted in higher turf quality and traffic tolerance than fast release fertilizer, however, shoot density did not showed a significantly different. For the fast recuperation of turfgrass from traffic injury in the early winter, it is recommended to avoid early morning and late evening traffic such as cup replacement and other maintenance practise. It is also recommended to delay the first tee-up time and ending early for last tee time during cold weather season.

• Korean Journal of Agricultural Science
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• v.8 no.2
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• pp.224-230
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• 1981

Threshing forces of seven varieties, that is, three Japonica type varieties-Irri 348, Irri 345, and Milyang 15, and four Indica type varieties-Milyang 23, Irri 342, Suweon 294 and Suweon 287-, which have been cultivated in the standard fertilization field being at Chungnam Provincial Office of Rural Development, Youseong, Chungnam-do, they were measured every other day in the period from September 28 to October 20 which were included the proper harves ting time. Also the threshing forces were checked in three-dimensional directions. Relationships between threshing forces and moisture contents of the unthreshed rice were examined in the laboratory as dried in the natural condition. The results of this study were as follows; 1. The mean threshing force of Japonica type varieties was about 1969r. and that of Indica type varieties was about 113gr. when external force was loaded in the direction parallel to the grain. 2. The threshing force bent to the grain was about 9.8through 28.2gr. equal to 7 through 21 percent to the threshing force parallel to the grain in Indica type varieties meanwhile about 59.8 through 115.0gr equal to 33 through 50 percent in Japonica type varieties. 3. Under the warmer temperature than $0^{\circ}C$, the change of threshing forces was not great in this experimental period on the harvesting dates. 4. The threshing force decreased s lightly as moisture content decreased in natural drying under the shade after cutting. 5. The threshing force of the lower portion was a little biger than that of the upper portion in an ear.

• 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.

• Journal of Embryo Transfer
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• v.14 no.1
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• pp.17-22
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• 1999

To investigate the maturational and development competece of porcine oocytes of different diameter groups, oocytes were obtained by aspiration from slaughterdhouse ovaries. After washing three times in NCSU23 medium, each cumulus-oocyte complex was transferred into a $8{mu}ell$ drop of the maturation medium (one oocyte per drop) under paraffin oil. The diameter without zona pellucida of oocytes was measured with micor-calibrator (Mikrometer, E. Leitz) on a screen connected to a VCR on an inverted microscope $(200\times)$. After being measured, the oocytes were divided into 6 groups according to their diameter size : <105, 105 to < 110, 110 to < 115, 115 to < 120, 120 to < 125 and > $125{\mu}{\textrm}{m}$, and in vitro maturation (IVM), fertillzation (IVF) and production (IVP) of oocytes / embryo was performed. The rates of in vitro maturation on oocytes in the greater 105 ${\mu}{\textrm}{m}$ size groups(91.8~100%) were significantly (P<0.05) higher than in the < 105 ${\mu}{\textrm}{m}$ group(66.7%). The rates of sperm penetration were significantly (P<0.05) low in < $105{\mu}{\textrm}{m}$ group (50.0%) than others groups (81.6~85.5%). But the plyspermic fertilization rate was significantly (P<0.05) higher in < $110{\mu}{\textrm}{m}$ oocytes groups than in the $110\leq{\mu}{\textrm}{m}$ size groups. The rates of cleavage and development to blastocysts rose as oocytes diameter increased, however, while oocytes over $120{\mu}{\textrm}{m}$ in diameter failed to develop to blastocysts. There results suggest that porcine oocytes have acquired full meiotic competece at a diameter of $105{\mu}{\textrm}{m}$ but not yet attended full development competence to blastocyst and that oocytes have acquired full development competence at a diameter of $110{\mu}{\textrm}{m}$.