• Journal of The Korean Society of Grassland and Forage Science
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• v.12 no.2
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• pp.104-110
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• 1992

This experiment was carried out to determine the effects of nitrogen(N) fertilization levels(0. 120, 240 and 360 kg/ha/y) on seasonal nitrate nitrogen($NO_3$-N) concentration and water soluble carbohydrate (WSC) content of grasses grown under shading condition. The plants were sampled on 7. 14, 21, 28 and 35 days after harvesting in spring(May), summer(July) and autumn(September), 1988. respectively. Shading degree was controlled artificially ca. 45-50%, and each annual level of N was distributed 5 times equally. The $NO_3$-N concentration of grasses was very low(0.064 %) in spring, high in summer(O.l98 %) and autumn(0.234%), and NO,-N was slightly decreased with growing of grasses. Also the NO,-N concentration of grasses was increased significantly with increasing level of N. In spring, the $NO_3$-N of grasses was very safety(less than 0.1 %), regardless of plant height and N level. In summer and autumn, however, plant heights of below 40 cm(summer) and 44 cm(autumn), and N levels of over 210 kg(summer) and 140 kg(autumn) were over toxic level to animals, respectively. The WSC content of grasses was slightly decreased with grass growth. and significantly decreased with high level of N. From the above results, it is suggested that 200 kg/ha of N in this experiment is effective for forage quality. Also a little more amount of N(50-70kgltimes) in spring. and low level of N(20-30 kgltimes) in summer and autumn season may be desirable on woodland posture.

• Journal of Embryo Transfer
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• v.9 no.1
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• pp.1-6
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• 1994

This experiment was investigated the effect of presence of granulosa cells from follicles of different size on bovine oocyte maturation, cleavage and development to late stage. The nuclear and cytoplasmic maturation of oocytes in the IVM-IVF system are critical for subsequent embryo development. Granulosa cells when the co-cultured with oocytes may interact with cumulus-oocytes complexes and influence the development competence of the oocytes. Granulosa cells from medium (2~6 mm) and large(>1O mm) size follicles were recovered by aspiration, washed 3 times by centrifugation at 500 x g for 5 min. and used for co-culture at a concentration of 2~3 x 106 cells/mi. The oocytes were matured in vitro (IVM) for 24 hrs. in TCM-199 supplemented with 35 $\mu$g/ml FSH, 10 $\mu$g/ml LH, 1 $\mu$g/ml estradiol-17$\beta$ and granulosa cells at 39$^{\circ}C$ under 5% $CO_2$ in air. They were fertilized in vitro (IVF) by epididymal spermatozoa treated with heparin for 24 hrs., and then the zygotes were co-cultured in vitro (I VC) with bovine oviductal epithelial cells for 7 to 9 days. The assessment of maturation revealed that Grade J oocytes showed significantly(P

• Environmental Analysis Health and Toxicology
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• v.25 no.3
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• pp.241-251
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• 2010

Crucian carp (Carassius auratus) has been used as the sentinel species for POPs (Persistent Organic Pollutants) monitoring in aquatic environment. However, there is little information for dioxin toxicity and especially, early life stage toxicity in crucian carp have been never carried out. In this study, we investigated several toxic effects for 2,3,7,8-tetrachlorodibenzo-p-dioxin (2,3,7,8-TCDD) in fertilized egg obtained by natural fertilization from crucian carp. The embryos at 3 h post-fertilization (hpf) were treated with 0.039, 0.156, 0.625, and 2.5 (${\mu}g/L$) TCDD by waterborne exposure for 60 minutes and changed with fresh water 2 times per day. Fertilized eggs started hatching at 51 hpf and TCDD exposed embryo showed decrease of hatching rate in a dose-dependent manner at 75 hpf. Pericardial edema was continuously observed in larvae exposed to TCDD from hatching start time (51 hpf), followed by the onset of mortality. In addition, AhR-related gene, CYP1A was clearly increased by TCDD in a dose dependent manner. These results indicated that fertilized eggs obtained from crucian carp have the TCDD related gene regulation and a distinct TCDD developmental toxicity syndrome by TCDD exposure. Therefore, we suggested that early life stage test in crucian carp could be used as test methods on dioxins toxicity.

• Development and Reproduction
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• v.25 no.4
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• pp.213-223
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• 2021

Controlled ovarian hyperstimulation (COH) is routinely used in the in vitro fertilization and embryo transfer (IVF-ET) cycles to increase the number of retrieved mature oocytes. However, the relationship between repeated COH and ovarian function is still controversial. Therefore, we investigated whether repeated ovarian stimulation affects ovarian aging and function, including follicular development, autophagy, and apoptosis in follicles. Ovarian hyperstimulation in mice was induced by intraperitoneal injection with pregnant mare serum gonadotropin (PMSG) and human chorionic gonadotropin (hCG). Mice subjected to ovarian stimulation once were used as a control group and 10 times as an experimental group. Repeated injections with PMSG and hCG significantly reduced the number of primary follicles compared to a single injection. The number of secondary and antral follicles increased slightly, while the number of corpus luteum increased significantly with repeated injections. On the other hand, repeated injections did not affect apoptosis in follicles associated with follicular atresia. The expression of autophagy-related genes Atg5, Atg12, LC3B, and Beclin1, cell proliferation-related genes mTOR, apoptosis-related genes Fas, and FasL was not significantly different between the two groups. In addition, the expression of the aging-related genes Dnmt1, Dnmt3a, and AMH were also not significantly different. In this study, we demonstrated that repeated ovarian stimulation in mice affects follicular development, but not autophagy, apoptosis, aging in ovary. These results suggest that repetition of COH in the IVF-ET cycle may not result in ovarian aging, such as a decrease in ovarian reserve in adult women.

• Korean Journal of Agricultural Science
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• v.49 no.1
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• pp.103-112
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• 2022

Fipronil is a popular insecticide used in both agricultural and domestic fields. Factors that affect sperm and eggs have a direct influence on reproductive outcomes. This study was undertaken to assess the effect of varying concentrations (10 - 200 μM) of fipronil and incubation times (30 min and 2 hrs) on boar spermatozoa. Spermatozoa were evaluated for motility, motion kinematics, viability, chromatin stability, and for the generation of intracellular reactive oxygen species (ROS) and the results were compared to those from corresponding controls. The findings revealed a significant, dose-dependent reduction in sperm motility in all fipronil treatment groups at 30 min of incubation (p < 0.05). A similar dose-dependent reduction in sperm motility was observed subsequent to fipronil exposure for 2 hrs of incubation (p < 0.05). Groups treated with fipronil showed a gradual reduction in motion kinematics (p < 0.05). Moreover, a significantly higher percentage of dead sperm was observed at 200 μM fipronil, as compared to the highest live percentage obtained in controls (p < 0.05). Evaluating the sperm chromatin integrity revealed a significantly higher percentage of damaged chromatin in spermatozoa incubated with 200 μM of fipronil. Moreover, ROS production was significantly higher in fipronil-exposed sperm (p < 0.05). In conclusion, boar spermatozoa incubated with fipronil showed decreased levels of sperm motility and viability, weaker chromatin integrity, and increased levels of intracellular ROS generation, all of which indicate that exposure to fipronil potentially impairs the fertilization competence of boar spermatozoa.

• Korean Journal of Agricultural Science
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• v.51 no.2
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• pp.109-121
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• 2024

Mancozeb is a manganese and zinc-containing fungicide that belongs to the ethylene bisdithiocarbamate group and produces ethylene thiourea (ETU) after biotransformation or environmental degradation, which has toxicological hazard owing to its known antithyroid properties. Although mancozeb leads to negative changes in fertility capacity, the effects of ETU are less known. Therefore, this study examined the alteration of fertilization competence in boar spermatozoa exposed to ETU. The sperm motility, motion kinematics, viability, acrosome integrity, chromatin stability, and intracellular reactive oxygen species (ROS) production of sperm subjected to various ETU concentrations (10, 50, 100, and 200 µM) were evaluated after two different incubation times (30 min and 2 hrs). In addition, the relative mRNA expression of the sperm functional proteins was analyzed after exposure to ETU. A dose-dependent motility reduction was observed in sperm exposed to ETU during both incubation periods compared to the controls. The motion kinematics were reduced significantly in sperm incubated with ETU. Higher percentages of viable sperm were observed in the controls, while such viability was decreased significantly in sperm with 10 - 200 µM ETU. The acrosome integrity was particularly damaged on sperm incubated with 10 - 200 µM ETU for 30 min. Higher intracellular ROS levels were produced in sperm exposed to 200 µM ETU. In addition, lower relative levels of AKAP3, AKAP4, ODF2, and ZPBP2 expression were observed in sperm exposed to ETU compared to the controls. Mancozeb and ETU could adversely affect the reproductive functions of mammals. Hence, the effects of ETU on the reproductive system should be examined further.

• Journal of the Korean Society of Tobacco Science
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• v.1 no.1
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• pp.14-20
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• 1979

This experiment was carried out to increase productivity of low productive fields (now producing less than 180kg per 10a ) at the Eumseong Tobacco Experiment Station in 1976- 1977. We could produce 220kg or more of yields (Hicks ) per 10a by increasing plant density to 2778 plants/10a and applying 20% more fertilizer than the recommended amount.

• Journal of Veterinary Clinics
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• v.17 no.2
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• pp.420-430
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• 2000

These studies were preformed to investigate the freezing conditions to achieve good post-thaw viability of spend and the practical methods of artificial insemination frozen canine semen. Semen were collected from nine male dogs which had been proved to be fertile in the past and the semen were treated for freezing procedure. Post-thaw motility and viability of canine sperm were evaluated to investigate individual tolerance of freezing, difference among freezing extenders, dif-ference among freezing equipments and freezing conditions, difference between fast and slow cooling rate, difference according to different glycerol concentration, effect of seeding on post-thaw viability, difference according to cutting part of straw, difference according to thawing temperatures, and dif-ference according to media added to thawed semen. Thawed semen for insemination were added with equal volnme of canine capacitation medium (CCM) and the volume of semen and the number per insemination were adjusted as 2-3 ml and $20-30 {\times}10^7,$ respectively. The semen were inseminated in vagina using balloon catheter and en17ryos were cellected from 9 to 11 days after the second Al to d determine fertilization.

• Journal of Embryo Transfer
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• v.9 no.1
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• pp.73-83
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• 1994

The objective of this study was to develop an effective in vitro production system capable of obtaining more porcine embryos from immature oocytes. These experiments were thus conducted to examine the effect of oocytes type and maturation time on the in vitro maturation(IVM) and fertilization(IVF) of oocytes and the in vitro development (IVD)of IVF embryos. 1. The degree of oocyte maturation based on cumulus expansion index(GEI) did not differ for A- and B-typed oocytes but the index of oocyte type C was lower(P<0.05) than that of other oocyte types. 2. When the oocytes of type A and B were matured for 36, 42 and 48hrs, the GEl was not different between the 36- and 42-h maturation but the GEl after 48hrs was greatly lower(P<0.05) than that of other maturation times. 3. The highest cleavage rate(48.6%) of IVF oocytes was obtained from A typed oocytes and 42-h maturation but the developmental potential based on cleavage index was the highest when B-typed oocytes were matured for 42hrs.

• Journal of Environmental Science International
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• v.23 no.5
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• pp.771-780
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• 2014

Tetragonia tetragonides is a medicinal plant native to ocean sand soil of southern provinces and has significant effects on the prevention and curing of gastroenteric disorders. Despite of its popularity, supply of the plant has never met the level of demand because of the absence of an adequate culturing method. The present study, thereby, was conducted for classifying the plants with geographically different characteristics, studying growth habits, developing a new culturing method and establishing a large scale propagation system of selected superior individual plants. The study was also aimed for revealing optimum conditions for seed treatment, fertilization, and efficient culturing system and thereby, for utilizing the plant as a new income source for rural communities. The seed was elongated with size of 2.6 mm (width) ${\times}$ 1.8 mm (length). No difference in seed size was observed depending on different inhabitate. Each flower produced about 4.5~4.8 seeds. Germination rate was high for seeds matured for 40 days after fertilization, but deceased to 50% for seeds matured only for 20 or 30 days. Seed dormancy lasted 6 months and seed storage at humid $5^{\circ}C$ facilitated germination. Mechanical obstruct of seed germination was due to seed coat and removal of seed coat enhanced the germination rate. Optimum temp. for seed storage was $5^{\circ}C$, and high germination rate was maintained for 350 days. However, for stratification condition or at room temperature, germination was significantly reduced as storage time increased Optimum treatment of plant growth regulators was soaking in $GA_3$ 250 mg/L for 1 hr. The priming treatment with 50 mM $Ca(NO_3)_2$ at $20^{\circ}C$ for two days improved the seed germination with 10% compared to non-treated control. The treatment of 20% NaOCl for 3 hr. improved the seed germination rate up to 10% and 1 day ahead.