• Proceedings of the Korea Society of Environmental Biology Conference
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• 2004.12a
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• pp.124-126
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• 2004

Molinate is thiocarbamate herbicide used primarily in rice production. Chondrogenesis is a multistep process that is essential for endocondral bone formation. The transcription factor Sox9 has an essential role during the sequential steps of chondrocyte differentiation. Bombina orientalis is one of the most common amphibians in the world and comprises a large proportion of their total number. We examined the embryotoxic and survival effects of molinate at various concentration in B. orientalis embryos. The survival rates of embryos at 312h post fertilization treated with molinate was decreased with concentration dependent manner. Also, developmental malformations appeared by molinate in B. orientalis embryos. the expression levels of Sox9 mRNA was examined by RT-PCR. In our result showed that Sox9 expression was found to be increased in malformed tadpole compared to normal tadpole. These results suggested that molinate was detrimental for survival and development of B. orientalis embryo.

• Journal of Ecology and Environment
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• v.30 no.3
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• pp.225-229
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• 2007

The mating systems of natural populations of Potentilla freyniana in Korea were determined using allozyme analysis. The result suggests that P. freyniana is outcrossing as well as employing vegetative reproduction by stolon (self-fertilization rate, s < 0.5). The values of the inbreeding coefficient of eight populations in Korea varied from 0.244 to 0.331, with an average value of 0.274. For eight natural populations, multi-locus estimates of outcrossing (tm) was 0.603 across 13 polymorphic loci, with individual population values ranging from 0.530 to 0.652. The relatively low outcrossing rates of some populations could be attributed to extensive vegetative reproduction by stolon and the isolation of flowering mature plants. Although P. freyniana usually propagated by asexually-produced ramets, I could not rule out the possibility that sexual reproduction occurred at a low rate because each ramet may produce terminal flowers. Although heterozygote excess was observed in some natural populations, most populations exhibited varying degrees of inbreeding and a heterozygote deficit.

• Journal of Aquaculture
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• v.10 no.4
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• pp.395-402
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• 1997

The study has been conducted in order to understand the inheritance of body color in the wild type zebrafish (zebra danio), Danio rerio, and its golden mutant (golden danio). The body color was also studied to determine the effect of golden coloration on the survival rate of zebrafish eggs and larvae up to 15 days after fertilization. Reciprocal monohybrid crosses between the wild and the golden type of zebrafish indicated that golden coloration was controlled by a single gene which had two alleles. Transmission of these alleles from parents to their progenies followed the principles of dominance and segregation based on Mendelian inheritance. Similar results from the reciprocal crosses implied that a locus for golden coloration was located on an autosomal chromosome. On the other hand, average survival rates from four different types of mating between, and within, zebra and golden danio suggested that golden coloration seemed to be associated with the survival rate of zebrafish, especially in its early embryonic stage. This indicated that homozygous recessive golden mutation was likely to weaken the golden danio's chance of survival.

• Asian-Australasian Journal of Animal Sciences
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• v.15 no.4
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• pp.485-493
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• 2002

Different factors may affect the sensitivity of porcine oocytes during cryopreservation. The effect of two methods (cooling and vitrification), four cryoprotectants [glycerol (GLY), 1, 2-propanediol (PROH), dimethyl sulfoxide (DMSO) or ethylene glycol (EG)] and two vitrification media (1 M sucrose (SUC)+8 M EG; 8 M EG) on the developmental capacity of porcine oocytes at the germinal vesicle (GV) stage or after IVM at the metaphase II (M II) stage were examined. Survival was assessed by FDA staining, maturation and cleavage following IVF and IVC. A toxicity test for different cryoprotectants (GLY, PROH, DMSO, EG) was conducted at room temperature before cooling. GV and M II-oocytes were equilibrated stepwise in 1.5 M cryoprotectant and diluted out in sucrose. The survival rate of GV-oocytes in the GLY group was significantly lower (82%, p<0.01) than that of the other group (92 to 95%). The EG group achieved a significantly higher maturation rate (84%, p<0.05) but a lower cleavage rate (34%, p<0.01) than the DMSO group and the controls. For M II-oocytes, the survival rates for all groups were 95 to 99% and the cleavage rate of the GLY group was lower than the PROH-group (21 vs 43%, p<0.01). After cooling to $10^{\circ}C$, the survival rates of GV-oocytes in the cryoprotectant groups were 34 to 51%, however, the maturation rates of these oocytes were low (1%) and none developed after IVF. For M II-oocytes, the EG group showed a significantly higher survival rate than those of the other cryoprotectant groups (40% vs 23-26%, p<0.05) and the cleavage rates of PROH, DMSO and EG group reached only 1 to 2%. For a toxicity test of different vitrification media, GV and M II-oocytes were equilibrated stepwise in 100% 8 M EG (group 1) and 1 M SUC + 8 M EG (group 2) or equilibrated in sucrose and then in 8 M EG (SUC+8 M EG, group 3). For GV-oocytes, the survival, maturation and cleavage rates of Group 1 were significantly lower than those in group 2, 3 and control group (p<0.05). For M II-oocytes, there were no differences in survival, maturation and cleavage rates between groups. After vitrification, the survival rates of GV and M II-oocytes in group 2 and 3 were similarly low (4-9%) and none of them matured nor cleaved after in vitro maturation, fertilization and culture. In conclusion, porcine GV and M II-oocytes do not seem to be damaged by a variety of cryoprotectants tested, but will succumb to a temperature decrease to $10^{\circ}C$ or to the process of vitrification, regardless of the cryoprotectant used.

• Clinical and Experimental Reproductive Medicine
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• v.19 no.2
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• pp.125-131
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• 1992

The possible effect of human follicular fluid(hFF) on the growth and development of fertilized oocytes and embryos is important because the fallopian tubes are exposed to FF after follicular rupture and the processes of fertilization and embryo cleavage occur inside the fallopian tubes. Previously, it was suggested that human FF might adversely affect on the development of early mouse embryos. In order to investigate the effect of hFF on the development of embryos, early mouse embryos were cultured in media containing various protein sources as bovine serum albumin(BSA), fetal cord serum(FCS) and FF. And we evaluated the development of early mouse embryos in terms of the morphology, cleavage rate, and cell count of blastcysts. There were no significant differences in the morula and blstocyst formation rates of 2-cell mouse embryos cultured in the media containg three different protein sources and three different concentrations of FF. The blastocyst formation rate of 1-cell mouse embryo cultured in FF group was significantly higher than that cultured in BSA group(P<0.05). The morula and blastocyst formation rates of 2-cell mouse embryos of the group cultured in the media containing FF were comparable with those of other two groups, in addition, the cell count of blastocysts of FF group in the 2-cell embryo culture was higher than those of BSA group and HCS group(P<0.01), and this finding was also noted in 1-cell embryo culture. There was no difference in the morula and blastocyst formation rates of the 2-cell mouse embryos cultured in the media containing different concentrations of FF. These results suggest that mature human follicular fluid has no inhibitory activity on the development of early mouse embryos even in high concentration and may be a good protein source which is positively associated with the development of mouse embryos in vitro especially in 1 cell embryo culture.

• Clinical and Experimental Reproductive Medicine
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• v.30 no.1
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• pp.57-64
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• 2003

Objective : We investigate the effects of multiple attempts of embryo transfer because of retained embryos in the catheter and of contaminated mucus on the transferred catheter. Materials and Methods: We respectively analysed data between November 1998 and August 2002 from 305 patients of 369 cycles who underwent IVF-ET. Of these patients, 47 patients of 50 cycles (Group 2) were required multiple trial of embryo transfer. They were compared with an age-matched control groups (Group 1) with female factor infertility. Pearson's $?^2$ and Fisher's tests were used to compare proportions between discrete variables. Noncategorical data were compared using t-test. Statistical significance was set at p<0.05. Results: Embryos were significantly more likely to be retained when catheter was contaminated with mucus (Group 1: 22.4%; Group 2: 44.0%). The clinical pregnancy rates, however, for the contaminated mucus or not, were 46.8%, 43.5% respectively. There was no significant difference clinical pregnancy rate between those who had all their embryos transferred at the first attempt (45.4%) and those who required more than one attempt (48.0%). Conclusions: Contaminated mucus in the catheter is associated with failed embryo transferred at the first attempt. Embryo transfers, however, that are repeated attempts do not adversely affect pregnancy rates following IVF-ET.

• Clinical and Experimental Reproductive Medicine
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• v.21 no.1
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• pp.69-76
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• 1994

The survival and pregnancy rates were compared between non-frozen embryos and cryopreserved embryos at either pronucleate or 2-4 cell stages using the freezing and thawing techniques being identical in both groups were compared with fresh embryos. 496 embryos were frozen with 1, 2-propanediol and sucrose and 117 2-4 cell stages embryos had been thawed and 79.6 and 66.0% of them respectively were survival. Clinical pregnancy rate was 19.2% for embryos frozen at the pronucleate stage and 19.0% for embryos frozen at the 2-4 cell stages while the pregnancy rate of non-frozen embryos was 21.3%. There were no significant difference in the survival and pregnancy rates of embryos frozen at pronucleate and 2-4 cell stages. The current cumulative pregnancy rate per retrieval in all cycles with frozen zygotes is 35.4 %, consid~ erably higher than observed in single transfers of embryos without cryopreservation(21.3%); predicted pregnancy rate after transfer of all frozen embryos is 43.3 %. It is concluded that firstly, the survival and pregnancy rate of cryopreserved embryos at pronucleate or 2-4 cell stages are very similar to those from their fresh embryos and non-frozen embryos and secondly, cryopreservation substantially enhances pregnancy attainment from in vitro fertilization.

• Korean Journal of Animal Reproduction
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• v.26 no.4
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• pp.311-320
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• 2002

The objective of this study was to assess the developmental potential in vitro produced embryos frozen-thawed with the various containers, and also examined expression and localization of heat shock protein 70 at these embryos. For the vitrification, 2-cell, 8-cell and blastocyst stage embryos produced by in vitro fertilization (IVF) and nuclear transfer (NT) were exposed the ethylene glycol 5.5 M freezing solution (EC 5.5) for 30 sec, loaded on each containers such EM grid, straw and cryo-loop, and then immediately plunged into liquid nitrogen. Thawed embryos were serially diluted in sucrose solution, each for 1 min. and cultured in CRI-aa medium. Survival rates of the vitrification production were assessed by re-expanded, hatched blastocysts. There were no differences in the survival rates of IVF using EM grid and cryo-loop. However, survival rates by straw were relatively lower than other containers. The use of cryo-loop resulted in only survival of nuclear transferred embryos (43.7%). Also, there embryos after IVF or NT were analysed by semi-quantitive reverse transcription-polymerase chain reaction (RT- PCR) methods for hsp 70 mRNA expression. Results revealed the expression of hsp 70 mRNh were higher thawed embryos than control embryos. Immunocytochemistry used to localize the hsp 70 protein in embryos. Two and 8-cell embryos derived under control condition was evenly distributed in the cytoplasm but appeared as aggregates in some frozen-thawed embryos. However, in the control, blastocysts displayed aggregate signal while Hsp70 in frozen-thawed blastocysts appeared to be more uniform In distribution. Therefore, this result suggests that the exploiting Hsp 70 in the early embryos may be role for protection of stress condition for increase viability of embryos within IVF, NT and there frozen-thawed embryos.

• Journal of the Korean Society of Environmental Restoration Technology
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• v.18 no.4
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• pp.1-14
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• 2015

A manipulation of zoysiagrass's lateral stems and soil-media; used for slope revegetation, is expected to facilitate the production of high-quality grass. To study the influences on the growth of zoysiagrass depending upon various soil-media conditions, two different types of soil are used. The results obtained - through investigation of its cover rates, leaf color and number - are summarized as follows. In mountain regions soil, there are no significant differences in growth and development of grass in treatments: zoysiagrass's lateral stems treatments with 1cm, 2cm, and 3cm soil-media and treatment with only seeding. Zoysiagrass, in most of the treatments, show about the same growth rates, and at the end, fair visual quality. Zoysiagrass's lateral stems treatments with 2cm, 3cm show slightly better growth, however, thickness of soil-media need not be more than 1cm to obtain an expected quality of lawn. In decomposed granite soil, there appears statistical significance in growth of the grass in treatments: zoysiagrass's lateral stems treatments with 1cm, 2cm, and 3cm soil-media and treatment with only seeding. The thicker the soil-media, the better the growth of grass, and that in treatment with seeding-only shows poor quality in general. And therefore, it is efficacious to plant in 3cm soil-media when quick formation of lawn is necessary; however, using 2cm soil-media is ultimately the most cost-efficient way of formation. But, when time allows - that is, more than three months - 1cm soil-media in decomposed granite soil is reasonable to formate just as effective lawn. And so when performing seeding, additional covering, fertilization or increasing the quantity of seed must be considered.

• Journal of Embryo Transfer
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• v.14 no.2
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• pp.113-119
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• 1999

This study was performed to improve the development of the in vitro fertilized bovine embryos by the condition of in vitro maturation. COCs were matured in TCM 199 supplemented with 0.1% PVA, 10ng/ml EGF, Hormones (5$\mu\textrm{g}$/ml FSH, 10 IU hCG, 1 $\mu\textrm{g}$/ml estradiol 17-$\beta$) or granulsa cell+Hormones atmosphere 39$^{\circ}C$, 5% CO2, 95% air for 24hrs. Matured oocytes were fertilized with frozen-thawed semen capacitated with 5mM caffein in BO medium for 20 hrs. IVF embryos were cultured in TCM 199 containing with hormones(same as matured medium), 10% FBS and co-culture with bovine oviduct epitherial cells. Maturation rates of COCs were showed 73.8%, 78.5%, 83.2% and 87.6% respectively, and were significant differences between PVA, EGF, and Hormones, GC+Hormones(p<0.05). The cleavage rates of IVF embryos were revealed 72.5%, 78.4%, 82.3% and 84.2% and showed same tendency as maturation rates(p<0.05). The blastocysts matured by above maturation condition and cultured for 7~10 days after fertilization had 34.4, 43.6, 52.3 and 59.3 cells had no differences among the treatments. These results suggest that high molecules as a substitutes of serum and growth factor may induce nuclear resumption of COCs but we need more study to produce transferable IVF blastocysts by use of that agents.