• Korean Journal of Animal Reproduction
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• v.22 no.1
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• pp.1-9
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• 1998

This study was carried out to confirm whether the developmental capacity of bovine mature oocytes frozen ultra-rapidly using electron microscopic(EM) grids and EFS30 can be obtained, and whether the cryoprotectants and the freezing method used in this study effect detrimentally to the bovine oocytes by indirect immunocytochemistry. As freezing solution, we used EFS30 which consisted of 30% ethylene glycol, 0.5 M sucrose, 18% ficoll and 10% FBS added in D-PBS. The results obtained in this experiment were summarized as follows: When the effects of cryoprotectant and freezing procedure on the microtuble, micrfilament and chromatin morphology of oocytes were evaluated using indirect immunocytochemistry, the results of freezing as well as exposure group were not different with that of the control oocytes. When the fertilization abnormality after ultrarapid freezing of bovine mature oocytes was examined by Hoechst staining, the rates of total penetration(96.7, 9.0%), normal two pronuclei formation(74.6, 68.9%) and mean number of sperm / oocyte(1.50, 1.44) were not different between control and freezing group. In addition, when the developmental capacity of frozen-thawed of oocytes(85.5%) was survived, 74.5% of them were cleaved and 31.4% of cleaved embryos were developed to blastocyst. These data were similar to those of the control(76.0%, 34.6%) and exposure(74.5%, 33.0%) except survival rates. Also, when the total cell number of blastocysts produced from the each treatment at day after IVF was examined by hoechst staining, there were not different among groups. There results demonstrate that developmental capacity of frozen-thawed bovine mature oocytes can be successfully obtained by ultra-rapid freezing method using EM grid and EFS30 solution.

• Korean Journal of Animal Reproduction
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• v.25 no.2
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• pp.155-162
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• 2001

These experiments were conducted to evaluate the effects of the administration of exogenous PG $F_{2{\alpha}}$ on hormone concentrations and follicular development in early postpartum(pp) Korean native goats. 1. Plasma PG $F_{2{\alpha}}$ concentrations in PG $F_{2{\alpha}}$ treated goals showed a gradual increase from a low on day 2 (GR-1 : 10 day-treatment group: 6.35$\pm$0.5 and GR-2; 4 day-treatment group: 0.22$\pm$0.2 pg/$m\ell$, respectively) to reach a peak of 21.18$\pm$1.6 or 4.21 $\pm$0.4 pg/$m\ell$ on day 4 after treatment of PG $F_{2{\alpha}}$. 2. Plasma PG $F_{2{\alpha}}$ concentrations in GR-1 goats averaged 9.08 $\pm$1.2 pg/$m\ell$ compared with 5.44$\pm$ 1.8 pg/$m\ell$ in GR-2 goats the day before treatment of PG $F_{2{\alpha}}$. Mean PG $F_{2{\alpha}}$ concentrations thereafter remained low during the treatment period but PG $F_{2{\alpha}}$ concentrations did not differ between the two group. 3. Plasma concentrations of estradiol-17 $\beta$ (E,) in PG $F_{2{\alpha}}$ - treated group were decreased gradually until 2 days after PG $F_{2{\alpha}}$ treatment but mean $E_2$ concentrations began to increase on 3 days and were Inaximal on the 4 days after treatment. 4. Plasma lulenizing hormone (LH) concentrations in PG $F_{2{\alpha}}$ - treated goats were slightly higher than in controls but mean LH concentrations did not differ between the two treatment groups. 5. Plasma prolactin (PRL) concentrations were suppressed in both GR-1(10 day-treatment group) and GR-2(4 day-treatment group) goats compared to saline controls but mean PRL concentrations remained lower in PG $F_{2{\alpha}}$ treated animals during post-treatment period. 6. The mean number of small and medium follicles present when PG $F_{2{\alpha}}$ was administrated was similar in all does but the increase in number of large follicles($\geq$4mm) tended to be higher in PG $F_{2{\alpha}}$ treated group than controls. These results suggest that concentrations of PG $F_{2{\alpha}}$ and estradiol-17$\beta$ were positively correlared with follicular diameter. We conclude that PG $F_{2{\alpha}}$ treatment stimulates follicular development similarly in both GR-1 and GR-2 group.

• Korean Journal of Animal Reproduction
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• v.21 no.3
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• pp.293-302
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• 1997

Follicular oocytes of Grade I and II were collected from 2~6 mm ovarian follicles and matured in vitro (IVM) for 24 hrs in TCM-199 su, pp.emented with 35$\mu\textrm{g}$/ml FSH, 10$\mu\textrm{g}$/ml LH, and 1$\mu\textrm{g}$/ml estradiol-17$\beta$ at 39$^{\circ}C$ under 5% CO2 in air. They were fretilized in vitro (IVF) by epididymal spermatozoa capacitated with heparin for 12 hrs. The zygotes were then co-cultured in vitro with bovine oviducted epithelial cells (BOEC) for 7 to 9 days. The optimal time for IVM, the successful enucleation of IVM oocytes by micromanipulation at different oocyte ages after IVM, and the ideal culture system for IVM for effective IVF and in vitro development of IVM-IVF embryos was examined for in vitro production of nuclear recipient oocytes and nuclear donor embryos. To improve the efficiency of nuclear transplantation (NT) of IVF embryo into IVM follicular oocytes, this study evaluated the optimal electric condition and oocytes age for activation of IVM oocytes and in vitro development of NT embryos. In vitro development of NT embryos with preactivation or non-preactivation in enucleation oocytes, cell number of IVN-IVF embryos, and NT embryos wre also examined. The results obtained were as follows; 1. The most suitable enucleation time was at 24 hpm (83.3%) rather than that of 28 hpm(69.6%) and 32 hpm(50.0%). 2. There was no difference among the fusion rates of NT embryos at the voltages of 0.75, 1.0 and 1.5 kV/cm, but the in vitro development rates to morule and blastocyst were significantly (P<0.05) higher at the voltage of 0.75(12.5%) and 1.0kV/cm (12.6%) compared to 1.5kV/cm(0%). 3. No significant difference in activation rates were seen in NT embryos stimulated for 30, 60 and 120 $\mu$sec (71.7, 85.2 and 71.9%, respectively), but the in vitro development rates to morulae and blastocyst were significantly (P<0.05) higher in the oocytes stimulated for 30 $\mu$sec (11.6%) and 60 $\mu$sec(10.7%) than 120 $\mu$sec(0.0%). 4. The fusion rates (71.0 and 87.3%) and the in vitro development rates (9.1 and 12.7%) to morula and blastocyst were seen in the NT embryos stimulated at 28 and 32 hpm under the condition of 1.0 kV/ml, 60 $\mu$sec. However, at 24 hpm the fusion rates were 64.8% and the in vitro development to morula and blastocyst were not seen. 5. The fusion rates between the 8~12, 13~17 and 18~22-cell stage of IVM-IVF embryos were not significantly different. The in vitro development rates of the fused embryos to morula and blastocyst which were received from a blastomere of 8~12, 13~17 and 18~22-cell stages of IVM-IVF embryos were 14.9, 8.3 and 6.5%, respectively. 6. The in vitro development rate of the enucleated recipient oocytes with preactivation (24.2%) to morula and blastocyst was significantly (P<0.05) higher than that of non-preactivation (12.8%). 7. The cell numbers of NT blastocyst and IVM-IVF blastocyst cultured during 7~9 days were 63$\pm$11 and 119$\pm$23, and then their the mean cell cycle number were 5.98 and 6.89, respectively.

• Korean Journal of Animal Reproduction
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• v.25 no.2
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• pp.163-169
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• 2001

OSBA(oocytes-sperm binding assay) is a tool developed for rapid test of optimal condition of IVF medium and protein source by binding ability of mouse sperm and egg. Mouse oocyte-cumulus complexes were prepared by removing of the cumulus cells with 0.1% hyaluronidase. 10$\pm$2 oocytes per 30 ${mu}ell$ medium drop were inseminated with 3 ${mu}ell$ sperm suspension and were cultured f3r 3 hours and 24 hours, respectively. And the oocytes were recovered gently and the No. of sperm bound on oocytes were counted. In the Exp. 1, the ratio of oocytes bound with one sperm at least were 60.2%(50/83), 2%(2/77) and 100%(79/79) in the medium with no protein, FBS(15%, v/v) and BSA(0.4%. w/v), respectively, Fetal bovine serum(FBS) seriously inhibited sperm binding on oocyte, although bovine serum albumin(BSA) promoted the binding ability. The inhibiting effect of FBS was dependent on the concentration of FBS. The sperm binding ability according to oocyte maturity was tested in the Exp. 2. There was no significant difference between Met. II (mature) and Met. I (intermediate mature) oocytes in the number of oocytes bound with sperm and the number of sperm bound on oocytes. Finally, in Exp. 3, two batches of Ham's F10 medium with good and poor quality by OSBA were tested (The ratios of embryos developed from PN 1-cell stage to hatched blastocyst; 25% vs. 70%). In the medium with good quality, sperm binding ability was significantly increased (P ＜ 0.05). The ratio of oocytes bound with one sperm at least was 66% and 90% in the medium with poor and good quality, respectively. Conclusively, It was possible to test IVF medium condition rapidly and easily by OSBA.

• Journal of Environmental Health Sciences
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• v.33 no.2 s.95
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• pp.123-131
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• 2007

Many environmental contaminants including several metals, polycyclic aromatic hydrocarbons, and pharmaceuticals, have been identified to be phototoxic in the water environment. Concerns regarding photo-enhancement of toxicity of several environmental contaminants have been increasing because of the increased level of ultraviolet irradiation on the earth surface. However, there exist arguments that there might be certain defense mechanisms taking place in the aquatic ecosystem, which may include behavioral characteristics or genetic acclimation. This study was conducted to understand the potential responses of aquatic receptors to several phototoxic metals in the real environment, where long-term acclimation of such organisms to low dose UV-B may take place. For this purpose, water flea Daphnia magna was acclimated to environmentally relevant dose of UV-B (12 to $18uW/cm^2$) for >11 generations. The differences in developmental and life history characteristics, and toxicity responses were evaluated. Acclimation did not affect the daphnids' growth, longevity, and reproduction characteristics such as time to first brood, and brood size: After 21 d, survival of D. magna was not influenced by UV-B acclimation. When the number of young per female was compared. the daphnids acclimated for 11 generations tend to produce less number of neonates than the un-acclimated individuals but with no statistical significance (p>0.05). Four metals that were reported to be phototoxic elsewhere were employed in this evaluation, that include As, Cd. Cu, and Ni. UV-B level being applied in acclimation did increase the toxicity of Cd and Cu, significantly (p<0.05). However, the toxicities of As and Ni were not affected by irradiation of UV-B. Phototoxic responses were evaluated between the acclimated and the un-acclimated daphnids. For Cu, UV-B acclimation led to reduction of the photo-induced toxicity $(p\approx0.1)$ in daphnids. Non-acclimated Daphnia were affected by 50% at 4.18 ug/l Cu. but UV-B acclimated individuals exhibited $EC_{50}$ of 5.89 ug/l. With Cd, UV-B acclimation appeared to increase phototoxicity (p>0.05). With As and Ni, UV-B acclimation did not influence photo-induced toxicity. This observation may be in part explained by the type of reactive oxygen species that were generated by each metal. Similar to UV-B light, Cu is known to generate superoxide anion by acting as redox cycling toxicant. This is one of the first studies that employed_laboratory based UV-B acclimated test species for photoenhanced toxicity evaluation.

• Korean journal of applied entomology
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• v.49 no.1
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• pp.61-67
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• 2010

Effect of inoculation level on pathogenicity, development, and propagation of entomopathogenic nematode, Steinernema arenarium was investigated using the last instar of great wax moth, Galleria mellonella. Pathogenicity of S. arenarium was higher with increasing inoculation level representing 82% at the rate of 5 infective juveniles (IJs) while >98% at the rate of >10 IJs. The number of IJs penetrated into the host was 2.7, 5.0, 7.4, and 12.2 at the rate of 5, 10, 20, and 40 IJs, respectively while 24.3 at the rate of 80 IJs and 40.2 at the rate of 160 IJs. Inoculation level did not affect female adult size (4,616 to 6,444 ${\mu}m$) while affected male adult size (1,600 to 1,934 ${\mu}m$). The rate of stunted female adults was 70.2% at the inoculation level of 80 IJs and 63.7% at the inoculation level of 160 IJs. The number of progenies was 20,431, 26,696, 47,943, 50,516, 58,701, and 74,235 at the rate of 5, 10, 20, 40, 80, and 160 IJs, respectively. The body lengths of IJs were different depending on inoculation level ranging from 636 to 1,496 ${\mu}m$.

• Journal of Life Science
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• v.26 no.4
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• pp.484-489
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• 2016

Living organisms can maintain or extend their territories by producing allelochemicals that influence the growth, survival, and reproduction of other organisms. To identify natural biostimulants of positive allelochemicals, we screened 18 common seaweed extracts for enhancement of rhizoid and blade production in a convenient Porphyra suborbiculata monospore assay. By addition of methanolic extract from the most potent green seaweed, Codium fragile, 100% and 50% enhancement doses reflecting the amount of C. fragile extract required to enhance rhizoid formation (in terms of number of spores with rhizoids per total spores tested) were approximately 100 and 50 μg/ml, respectively, in the P. suborbiculata monospore culture. The C. fragile extract quickly enhanced rhizoid formation, rhizoid numbers per rhizoid-holding spore, rhizoid length, blade formation (in number of spores with blade per total spores tested), and blade length from most monospores at early culture days. The extract enhanced rhizoid formation after 2 days of culture significantly, rhizoid numbers per rhizoid-holding spore after 3 days, rhizoid length after 3 days, blade formation after 2 days, and blade length after 1 day, respectively, from most monospores. The allelochemicals that enhanced favorite seaweed species may be efficacious for new seaweed management technologies, including the development of biostimulant agents based on natural products.

• The Korean Journal of Ecology
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• v.27 no.5
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• pp.257-261
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• 2004

Juniperus chinensis var. sargentii Henry is a short and creeping evergreen shrub which reaches about 50㎝ in height and occurs in the northeast Asia and in high mountains over the South Korea. Its distribution is restricted, and the number of individuals are gradually decreasing. This study was conducted to estimate spatial pattern, genetic diversity and spatial genetic structure of J. chinensis var. sargentii. A total of 131 clumps were studied in the study area (40m × 60m). The spatial pattern of this population was random (Aggregation index R=1.031). In spite of the small number and the limited distribution, the level of genetic diversity (Shannon's index 1=0.463) was relatively high as compared with those of other plant species with similar ecological characteristics. ISSR genotypes of all individuals were investigated to find the genetic relationship of clumps and genets. Fifteen clumps were composed to be clones, and a total of 116 unique genotypes were composed by separate genets. Spatial autocorrelation analysis using Tanimoto distance showed that the genetic patch was established within 8m. The effect of clonal reproduction on genetic structure was almost nothing.

• Parasites, Hosts and Diseases
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• v.23 no.1
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• pp.1-6
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• 1985

Five cases infected with Rhabditis sp. were detected in a survey to examine the stool specimens from rural primary school children. A large number of the larvae of Rhabditis sp. detected by the direct cellophane thick smear were cultured by the alter paper method. The eBamination was carried out in April through June, 1980 in Tangjeong-Myon, Ahsan.Gun, and in September 1983 in Sandong-Eup, Yeongwol-Gun. The results obtained in this study were as follows: 1. Prevalence: Out of 925 children, 5(0.54%) children were found to be positive. The number of the detected larvae was 110/0.1gm of feces in case 1, 35 in case 2, 130 in case 3, 86 in case 4 and 62 in case 5. 2. Larvae: The larvae were prepared by means of the direct fecal smear and measured by a micrometer equipped in the microscope. Twelve (12) through 15-day old larvae in culture were $197.1{\mu}{\textrm{m}}$ long in average, and the maximum size of the matured stage larvae was $884.0{\times}25.9{\mu}{\textrm{m}}$. However, the length variation was ranged as 173.0 to $884.0{\mu}{\textrm{m}}$. 3. Adults: The size of clubbed adult female was $1, 357{\mu}{\textrm{m}}(1, 176~1, 419)$ in length and $80{\mu}{\textrm{m}}(79~82)$ in width. Length of buccal cavity was $33{\mu}{\textrm{m}}$. A long cylindrical esophagus($273{\mu}{\textrm{m}}$) of the worms with a valved posterior cardiac bulb and with median bulbar swelling was morphologically indicated. Ditance from mouth to vulva was occupied 58% of body length. Male worm was $1, 006{\mu}{\textrm{m}}(890~1, 148)$ in length and $49{\mu}{\textrm{m}}(48~49)$ wide. Caudal alee of bursa and spicules ($75{\mu}{\textrm{m}}$ in length) were well developed. 4. Eggs: The oval shaped eggs in the female uterus, when cultured, were $66{\times}56{\mu}{\textrm{m}}$ in sixte, and the eggs laid by the adult could not be detected. So, reproduction might be thought to be ovoviviparity. 5. The five cases were re-examined during the period from the 1st to the 3rd week after stool examinations, but Rhabditis sp. were detected again in 4 cases in 1st week. When they were examined in 3rd week, larvae could not be detected. So, it was thought that the infection of Rhabditis sp. to humans was facultative.

• Development and Reproduction
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• v.15 no.2
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• pp.133-141
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• 2011

The marine ornamental industry has become a multi-billion dollar industry these days. As developing, however, this industry has been criticized for the indiscriminate captures and the destruction of the surrounding environment. To circumvent these problems, it is suggested to breed the organisms artificially. While clownfishes Amphiprion sp. and Premnas sp. are the most famous ornamental organisms in the trade, few studies are yet available on the culture and commercial production of these fishes. These studies were performed to investigate the spawning periodicity, behavior and the habits during egg incubation, and to provide the information on the mass hatching system. The spawning periodicity and frequency were different in 4 pairs under the constant condition, temperature, salinity and photoperiod. On the contrary, the male's behaviors for egg incubation are almost same in the all. The egg-fanning activity of the male increased as the developing eggs reaching to the hatching day. Based on the above results, we designed a new artificial hatching system, the rotating type (RT), and compared it with the aeration type (AT) and spray type (ST) that were previously described. RT showed higher hatching rate of 87.3% than AT (74.4%) and ST (60.5%). Also, there were no significant differences in the hatching rate regardless of the number (2, 3, 5) of hatching plates. We suggest RT may accommodate various number of hatching plates and constitute a better hatching system for clownfishes.