• The Korean Journal of Malacology
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• v.19 no.2
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• pp.107-110
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• 2003

The effect of broodstock age on the spawning performance of cultured abalone, Haliotis discus hannai, was assessed for five age groups (3, 4, 5, 6, 7 years old). Spawning performance of the five age groups was assessed in terms of the percentage of spawning abalone, eggs per spawning female, fertilization rates, hatch rates and attachment rates per spawning. The percentage of spawning abalone increased along with broodstock age, reached maximum in female groups of 5- and 6-year age (60%), and in male group of 6-year age (100%), subsequently decreased in 7-year age group. The number of eggs produced per spawning also increased with broodstock age, and the maximum was found in 6-year age. Although the number of spawned eggs for the oldest abalone declined, it still spawned much more eggs than 3, 4, and 5 years old females. The fertilization and hatch rates were obviously larger for 4-6 years old group than the younger and older abalones. The maxima were found in 5-year age group (96.0% and 84.5%), and the minimum were appeared in 7-year age group (79.3% and 58.2%). The attachment rate increased with broodstock age, reached maximum in 6-year age group (33.6%), although the increment gradually declined. The attachment rate for 7-year age group rapidly decreased (16.6%). These results suggested that broodstock age affected the spawning performance of H. discus hannai, which peaked between 5 and 6 years old, and broodstock should be bred during this period for hatchery production.

• Clinical and Experimental Reproductive Medicine
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• v.27 no.1
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• pp.23-29
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• 2000

Objective: Zona pellucida (ZP) has been thought to be the barrier of egg to sperm penetration before and after fertilization. The phenomenon of ZP hardening has been considered as a post-fertilization event until now, and it is generally accepted that it is caused by the secretory products of cortical granules released during the cortical reaction. Hardening of ZP could occur "spontaneously" in mammalian oocytes in standard culture conditions, and that it is probably not a consequence of cortical reaction. The purpose of our study was to investigate the effect of human amniotic fluid (HAF) on nuclear maturation (NM) and fertilization ability of mouse immature oocytes. Methods: HAF was obtained from patients undergoing amniocentesis at $16{\sim}20$ weeks of gestation. HAF from five to ten patients was centrifuged and the supernatants was pooled. Cumulusenclosed mouse immature oocytes were incubated in the medium containing HAF, and examined to confirm NM and fertilization. Female ICR mice (about 3 weeks old) were stimulated with 7.5 IU PMSG. Immature oocytes were isolated at $48{\sim}52$ hrs post PMSG injection and cultured in TCM-199 supplemented with 20% HAF for 18 hrs. FBS was used as a control for the examination. Matured oocytes (MII) were fertilized with sperms collected from the epididymis of male mice (over 10 weeks old). Fertilization was in conducted T6 medium containing 15 mg/ml BSA, and confirmed at 6 hrs post-insemination. Fertilization rate was assessed in zona-intact or zona-free oocytes (denuded by trypsin). Evaluation of NM and fertilization was carried out by rapid staining method. ZP hardening was evaluated by incubating cumulus cell-free mature oocytes in 0.001% chymotrypsin at $37^{\circ}C$ for 10 min. Results: There was no significant difference between the effects of HAF (86.6%) and FBS (87.7%) supplements on NM of immature oocytes. When maturation medium was supplemented with HAF, total fertilization rates (7%) were significantly lower (p<0.01) than that of FBS (85.1%). In HAF group, fertilization rate was increased (p<0.01) in zona-free oocytes (7% versus 100%). The resistance of mouse oocyte ZP to digestion by chymotrypsin after maturation in vitro was significantly higher (p<0.01) in HAF group (86.7%) than in FBS (6.7%). To culture oocytes in FBS were very effective in preventing ZP hardening. However cultured oocytes in HAF showed high rate of ZP hardening (p<0.01). Conclusions: These results suggest that HAF can be used as a supplement for the NM of mouse immature oocytes in vitro. However, HAF induces spontaneous hardening of ZP of mouse immaure oocytes during maturation in vitro.

• The Korean Journal of Zoology
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• v.28 no.2
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• pp.71-84
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• 1985

Syntheses of RNA and protein were studied to examine changes in activating stored mRNAs during the early development of a sea urchin, Hemicentrotus pulcherimus. The rates of RNA and protein syntheses are very low in the unferilized eggs but the protein synthesis is activated upon the fertilization, while RNA synthesis remains still inactive at the same stage. These rates increase drastically at the blastula and gastrula stages, although the increases are not exactly in parallel. The protein synthesis was found to be also changing in quality during the early development from the studies by the two-dimensional gel electrophoresis.

• KOREAN JOURNAL OF CROP SCIENCE
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• v.34 no.4
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• pp.341-344
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• 1989

The relationship between the rates of rice grain shattering caused by typhoon and characteristics considered to be related with grain shattering such as heading date, amount of grain production and ethylene evolution of rice plant grown under the different methods of fertilization was analyzed. The grain shattering rates which was traced by the fallen grains after typhoon 'Dinah' occured in August 28 to 29, 1987, in the plots with nitrogen such as nitrogen alone, NP, NK, NPK, NPK + SiO$_2$ and NPK+compost plot etc. was lower than that in without nitrogen plots such as no fertilizer, compost alone, PK, P and K alone etc. and the amount of ethylene evolved from the leaf blades also showed the same trends. The correlation between the grain shattering rate and grain yield was negative but ethylene evolution was positively correlated with grain shattering rate of rice plant.ice plant.

• Korean Journal of Organic Agriculture
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• v.12 no.2
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• pp.209-217
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• 2004

The aim of this experiment was to investigate the effects of applying times and dilution rates of cattle slurry on dry matter yields of orchard grass. Cattle slurry was applied at the rates of average mineral nitrogen fertilizer equivalent to 150 kg/ha/year in 3 cutting frequency. Significantly higher dry matter yields than that of no fertilization (3.04 ton DM/ha) were recorded in the application of diluted cattle slurry(5.38~6.25 ton DM/ha) (p<0.05). Especially, this tendency was shown with higher annual dry matter yields at the partitioned dressing times, such as at the applications for 1st and 2nd growth, 1st and 3rd growth, and 1st, 2nd and 3rd growth respectively. The yields of annual dry matter both at fertilizing phosphorus and potassium, and phosphorus, potassium and nitrogen were higher than no fertilization as 5.41 tons and 8.78 tons per ha respectively. However, with application of diluted cattle slurry, dry matter yield par year (5.84 ton DM/ha) was higher than those of fertilizing phosphorus and potassium. The efficiencies of dry matter production with mineral and cattle slurry nitrogen application (kg DM/kg N) were 225 and 15.8~18.7 kg DM/kg N respectively. Especially, these tendencies were higher in diluted application plots than in no-diluted application plots of cattle slurry.

• The Korean Journal of Ecology
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• v.26 no.2
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• pp.71-74
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• 2003

Effects of nitrogen addition on the growth of Indigofera pseudo-tinctoria (Leguminosae) in the waste landfill site was investigated. Nitrogen fertilization in the nitrogen poor soils of waste landfill may influence the growth of nitrogen fixing plants beneficially or detrimentally. When I. pseudo-tinctoria was fertilized with three different levels of nitrogen, the coverage of plants treated with 46 g N/$m^2$ and 460 g N/$m^2$ was significantly less than that of plants treated with 23 g N/$m^2$. The growth rates of plant height treated with 46 g N/$m^2$ and 460 g N/$m^2$ were significantly less than those of plants treated with 23 g N/$m^2$. The growth rates of plant diameter treated with 46 g N/$m^2$ and 460 g N/$m^2$ were significantly less than those of plants treated with 23 g N/$m^2$. Dry weights of whole plants in control sites were higher than those of all the others nitrogen treatment sites. Nodule numbers were higher in control plants than those of plants in all the other nitrogen treatment sites. It is suggested that nitrogen fertilizer addition over 23 g N/$m^2$ affect the growth of some nitrogen fixing plants, such as I. pseudo-tinctoria, negatively.

• Clinical and Experimental Reproductive Medicine
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• v.41 no.1
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• pp.9-14
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• 2014

Objective: The aim of this study was to evaluate the efficiency of the intracytoplasmic morphologically selected sperm injection (IMSI) technique compared with conventional ICSI and previous ICSI attempts in oligo-astheno-teratozoospermia (OAT) patients. Methods: The sperms were selected under high magnification ($6,600{\times}$) and used to induce fertilization in previous ICSI patients by IMSI. These results were compared with previous conventional ICSI cycles in patients with OAT infertility. Results: These results demonstrated no significant difference in the fertilization rate between IMSI and previous ICSI cycles (67.7% vs. 65.0%). However, the pregnancy and implantation rates with IMSI were significantly higher than those of the ICSI cycles (33.3% vs. 12.5% and 14.6% vs. 5.4%, respectively; p<0.05). The miscarriage rate among pregnant patients (18.2% vs. 37.5%) showed no statistically significant difference between groups. Conclusion: Compared to conventional ICSI, this study found that IMSI increased the IVF-ET success rates in patients with OAT.

• Asian-Australasian Journal of Animal Sciences
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• v.13 no.6
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• pp.739-747
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• 2000

This experiment was conducted to evaluate the influence of dimethyl-sulfoxise (DMSO, 0, 5, 50, 100 and $500{\mu}M$) on the motility and acrosome reaction of the frozen-thawed spermatozoa from 3 different bulls (Bull A, Band C). Also we evaluated the developmental capacity of bovine embryos fertilized in a medium containing DMSO at various concentrations. DMSO had negligible effects on the sperm motility and acrosome reaction in all three bulls. However, the development rates from 2 to 16 cells stage on the 3rd day after insemination with 50, 100 and $500{\mu}M$ DMSO in Bull-B, and up to the blastocyst stage fertilized with 5, 50, 100 and $500{\mu}M$ in Bull-A were significantly higher (p<0.05) than those of control ($0{\mu}M$ DMSO) group from each bull. Furthermore, the rates of blastocysts per cleaved embryos of 5 to $500{\mu}M$ DMSO group in Bull-A and of 5 to $100{\mu}M$ DMSO in Bull-C were also significantly higher (p<0.05) than those for their $0{\mu}M$ groups, respectively. These results indicate that DMSO at micromol level used for in vitro fertilization might stimulate the development of embryos for some bulls.

• Reproductive and Developmental Biology
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• v.29 no.3
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• pp.141-144
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• 2005

This experiment was to investigate whether the mitochondria function assessment can be used for the prediction of sperm fertility through examining the correlation between mitochondria fluoromicroscopic frequency of frozen-thawed eight Hanwoo bull semen using rhodamine123 (R123) and in vitro embryo development following fertilization. Individual sperm were stained in 5 ${\mu}g/mL$ R123-added calcium-free Sp-TALP for 30 min at 0 h, 6 h, 12 h and 24 h after thawing and examined their mid-piece under an epifluorescence microscope using 495 nm excitation filter (x1,000). Three replications were taken, and at least 300 sperm per individual were examined. When semen samples were separated into two groups (good and poor) by sperm motility and fluorescent frequencies at just after thawing, average fluorescent frequencies were remarkably reduced as time going (0 h; $53.29{\~}72.94\%$, 6 h; $21.40{\~}58.90\%$, 12 h; $8.26{\~}25.93\%$, 24 h; $1.00{\~}13.78\%$, irrespective of selected group, and there were no differences at 6 h or 12 h after thawing between selected groups but indicated significant difference at 24 h after thawing (p<0.05). In vitro fertilization rates in good and poor groups ranging $70.8{\~}77.8\%$ and $52.1{\~}84.5\%$, respectively, were not significantly different. However, in vitro development rates of the same groups ranging $25.7{\~}40.0\%$ and $12.9{\~}1.8\%$, respectively, were significant different (p<0.05). These results demonstrate that mitochondria fluoromicroscopic assessment of frozen-thawed bovine sperm may be used as a criterion to select more fertile sperm.

• Journal of Embryo Transfer
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• v.25 no.2
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• pp.97-101
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• 2010

Intracytoplasmic sperm injection (ICSI) is one of the artificial fertilization methods when only a few sperm are available for insemination, and an important tool for the preservation of genetic materials of endangered animal species, especially the male is infertile. Different from other species such as mice and pigs, the conventional ICSI method which uses spiked pipette for injection (Spike-ICSI) is exhibited low success rates in cattle because the bovinesperm head membrane is hard to break during injection procedure. We chose piezo-assisted ICSI (Piezo-ICSI) for the improvement of the injection procedure including sperm head membrane rupture and efficient puncture of the plasma membrane of the oocytes. In this experiment, we compared the efficacy of the bovine ICSI embryo production between the Piezo-ICSI and Spike-ICSI. The second polar body extrusion, pronuclear formation, cleavage and blastocyst formation were evaluated after implementation of two different ICSI techniques. The Piezo-ICSI tended to show comparably higher rates of the second polar body extrusion (41.7%), the pronuclei formation (42.9%) and the two-cell cleavage (41.4%) than Spike-ICSI does (33.3%, 28.6% and 23.5%, respectively) although there is no statistic significance between two groups. In addition, the blastocysts were only obtained from the Piezo-ICSI group (10.3%). Our finding shows that the Piezo-ICSI may be used as an artificial fertilization method in cattle when in vitro fertilization is not applicable.