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.
This study was carried out to improve of effective culture system on development of IVM/IVF/IVC bovine embryos. The cumulus-oocyte-complexes (COCs) collected from Korean cattle ovaries harvested at a local abattoir were matured in 50 ${mu}ell$ of TCM199 supplemented with 10% fetal bovine serum (FBS) and hormones (35 $\mu\textrm{g}$/$m\ell$ FSH, 10 $\mu\textrm{g}$/$m\ell$ LH, 1$\mu\textrm{g}$/$m\ell$ estradiol 17 $\beta$ under paraffin oil at 39$^{\circ}C$ in a humidified atmosphere of 5% $CO_2$in air. At 24 hrs after culture, matured oocytes were fertilized in vitro for 22~24 hrs with motile semen in which obtained by centrifugation of a frozen thawed semen on Percoll-density gradients (45% vs. 90%) at 500 g for 20 min. The presumptive zygotes were divided into three experimental groups. Single egg (Group 1), 25 (Group 2) or 50 eggs (Group 3) were cultured on cumulus cell in 50 ${mu}ell$ TCM199 supplement with 10% FBS for 6~9 days after fertilization. In vitro developmental rates into the blastocysts in the groups 2 and 3 were significantly (P<0.05) higher than those of group 1 (37,27 vs. 6%, respectively). Cell number of blastocysts obtained in groups 2 and 3 at day 8 were significantly (P${mu}ell$) resulted in higher developmental competence and cell number of bovine blastocysts produced in vitro than those the culture of single embryos with cumulus cells.
This study was performed to investigate the reproductive characteristics of the cloned Hanwoo bulls produced by SCNT. The semen ejaculated from the cloned bulls (C-38 and C-39) and normal Hanwoo bull was properly measured the volume, the number of sperm, and the viability of frozen-thawed sperm. The sperm activity was analyzed using computer assisted sperm analysis (CASA). To analyze fertilizing ability of the cloned bulls, in vitro fertilization and artificial insemination were performed using the frozen-thawed semen. There were no differences in semen volume, sperm concentration, and the viability of frozen-thawed sperm between cloned bulls and normal bull. The difference was statistically significant in total motility, curvilinear velocity (VCL), straight-line velocity (VSL), and average-path velocity (VAP) of both cloned bulls compared to those of normal Hanwoo bull, respectively (p<0.05). The cleavage and blastocyst development rate were not different between the groups. five cloned cows were artificially inseminated using the frozen-thawed semen of C-38, two of them became pregnant. Two second generation calves (one male and one female) were produced. Based on these results, the cloned Hanwoo bulls showed normal reproductive abilities of semen parameters and sperm activity to their comparators and produced cloned calves, although there are some individual differences on the parameters.
Data from Hanwoo steers and bull calves were analyzed to see the phenotypic and genetic relationships between carcass traits from four different covariance models. Four models fit test station and test period as fixed effect of contemporary group and sire as random effect assuming paternal half-sib relationships among animals. Each model fits one of linear covariate (s) of different slaughter end points-age at slaughter in the first order, age at slaughter in the first and second order, slaughter weight or back fat thickness at 12-13th rib of cold carcass. Age at slaughter in its second order was not significant. Age at slaughter accounted for signifi- cant amount of genetic variances and covariances of carcass traits. Heritability estimates of back fat thickness, rib eye area, carcass weight, marbling score and dressing percentage were 0.34, 0.22, 0.24, 0.42 and 0.18, respectively at constant age basis. The genetic correlation between carcass weight and the other variables were all positive and low to high in magnitude. Genetic correlations between back fat thickness and rib eye area and between marbling score and dressing percentage were low but negative. Variance and covariance structure between these traits were shifted to a great extent when these variables were regressed on slaughter weight or on back fat thickness. These two covariates counteracted to each other but they adjusted each carcass variable or their interrelationship according to differential growth of body components, bone, muscle and fat. Slaughter weight tended to decrease genetic variances and covariances of carcass weight and between component traits and back fat thickness tended to increase those of rib eye area and between rib eye area and carcass weight.
Adiponectin is adipocyte complement-related protein which is highly specialized to play important roles in metabolic and honnonal processes. This protein, called GBP-28, AdipoQ, and Acrp30, is encoded by the adipose most abundant gene transcript 1 (APM1) which locates on human chromosome 3q27 and mouse chromosome 16. In order to determine chromosomal localization of the porcine APM1, we carried out PCR analysis using somatic cell hybrid panel as well as porcine whole genome radiation hybrid (RH) panel. The result showed that the porcine APM1 located on chromosome 13q41 or 13q46-49. These locations were further investigated with the two point analysis of RH panel, revealed the most significant linked marker (LOD score 20.29) being SIAT1 (8 cRs away), where the fat-related QTL located. From the SSCP analysis of APM1 using 8 pig breeds, two distinct SSCP types were detected from K~ native and Korean wild pigs. The determined sequences in Korean native and Korean wild pigs showed that two nucleotide positions (T672C and C705G) were substituted. The primary sequence of the porcine APM1 has 79 to 87% identity with those of human, mouse, and bovine APM1. The domain structures of the porcine APM1 such as signal sequence, hypervariable region, collagenous region. and globular domain are also similar to those of mammalian genes.
Geochemical approaches on the two recovered piston cores were performed to understand the characteristics of organic matters and the influence of the sea level variation of the East Sea in the Hupo Basin since the Holocene. The analyzing results on organic components (TOC and TN), and isotopic compositions of organic matters showed the variation to core locations and sampling depths. In core 08HZP-01, their values were gradually changed with depth from 4 mbsf to seafloor. However, rapid variation was observed at the boundary of 4.71 mbsf (meter below seafloor) in core 08HZP-03. Based on TOC/TN, $\delta^{13}C_{org}$ and $\delta^{15}N_{org}$, the origin of organic matters in the Hupo Basin can be divided into three groups; 1) predominant marine algae, 2) $C_3$ land plant, and 3) mixture of $C_3$ land plant and marine/freshwater algae. It is likely that the vertical and spatial variations of organic and isotopic compositions reflect the shifts in sedimentary environment (including sediment transportation) by ocean currents and sea-level changes and others during the Holocene period.
Seasonal variations of dominant algae species and the effects of these algae on coagulation and filteration of water treatment were investigated at Chilseo water treatment plant in downstream of Nakdong river from January in 1995 to Desember of 1998. The water quality of Nakdong river was found to be a hyper eutrophic state during the investigation periods. In the measurement, Chlorophyll-a contents ranged $20.7{\sim}180.9{\mu}g/l$ and total nitrogen contents(T-N) and total phosphorus contents(T-P) exceeded more than 3.4mg/l and 0.1mg/l, respectively. The changes in dominant algae species was in the order of Stepanodiscus sp., Asterionella sp., Melosira sp., Microcystis sp. and Synedra sp. from spring to winter. Microcystis sp. especially, was blooming during summer and Synedra sp. and Stepanodiscus sp. during winter. Although most diatomous algae appeared in the water treatment process caused filter clogging and reduced efficiency of coagulation and sedimentation, Synedra sp. and Stepanodiscus sp were revealed as the main trouble algae. Malfunction of water treatment process caused by Synedra sp. and Stepanodiscus sp. started at the algae concentrations of 800cells/ml and 1,820cells/ml, respectively. When chlorophyll-a content was $18.9{\mu}g/l$, the optimum amounts of coagulant were found to be 40mg/l of Alum and 16mg/l of PACS. Under condition of chlorophyll-a content of $154.1{\mu}g/l$, addition of Alum at the level of 75mg/l and PACS at the level of 35mg/l showed the lowest turibidity. The result indicates that increased amounts of the coagulants should be added for a better water treatment as chlorophyll-a contents increased. Addition of Alum at the amount of 60mg/l and 30mg/l of PACS removed Stepanodiscus sp. algae at the rate of 85% and 83%, respectively. In case of Synedra sp., 50mg/l of Alum and 25mg/l of PACS showed removal rates of 79% and 81%, respectively. Synedra sp. algae at the standing crops of 1,500cells/ml started filter clogging and a filtering process was completely inhibited after 8 hours. At this situation the filter clogging by Synedra sp. algae occurred at the depth of 5cm from the top anthracite layer. On the other, other algae did filter clogging at the depth of 10cm.
Because the household garbage had high water contents, it is difficult to compost it without an additive. Therefore, this study was performed to investigate possibility of using the waste newspapers as a humidity conditioner for the household garbage composting. The maximum temperature was $66.0^{\circ}C$ in spring, $69.2^{\circ}C$ in summer, $60.9^{\circ}C$ in fall and $56.0^{\circ}C$ in winter for composting periods. The seasonal pH value reached around 8.5 after 1 week and then repeated fluctuation at the narrow range in spring and fall, while it was stabilized at the range of $8{\sim}9$ after increasing to 8.5 after 1 week in winter. The water content was reduced little in winter, while decreased significantly in the other seasons. The water content after 8 weeks was 22.2% in spring, 47.6% in summer, 25.5% in fall and 72.5% in winter. The mass was reduced rapidly during the first week of each season, but it did not show much decrease. The volume reduced after 8 weeks to 59%, 32%, 27%, and 34% in spring, summer, fall and winter respectively. Organic matter content decreased over the four seasons. Nitrogen contents were in the range of 0.7% to 2.2% during the four seasons. The contents of inorganic compounds based on dry matyter were in the range of $0.94{\sim}2.59%\;P_2O_5$, $1.23{\sim}1.87%\;CaO$, $0.37{\sim}0.46%\;MgO$, $0.55{\sim}1.98%\;K_2O$. Concentration of heavy metals(Hg, Cd, Pb, Cu, Cr, Zn, As) based on dry matter were less than the limiting value of the by-product compost.
A model of pond system is developed for treatment and recycling of excreta from twenty-five adult dairy cattle. It is composed of wastewater treatment ponds and small fish ponds. Those are three facultative ponds in series; primary-secondary-tertiary pond and these are designed to rear carps without feeding. A pit is constructed at the bottom of primary pond for efficient sludge sedimentation and effective methane fermentation. It is contrived to block into it the penetration of oxygen dissolved in the upper layer of pond water. The excreta from the cattle housed in stalls are diluted by water used for clearing them. The washed excreta flow into the pit. The average yearly $BOD_5$ concentration of influent is 398.7mg/l. That of the effluent from primary, secondary and tertiary pond of the system is 49.18, 27.9, and 19.8.mg/l respectively. Approximate 88, 93, and 95 % of BOD5 are removed in each pond. The mean yearly SS concentration of influent is 360.5 mg/l That of the effluent from each pond is 53.4, 45.7, and32.7mg/l respectively. Approximate 86, 88, and 91% of SS are removed in each pond. The $BOD_5$ concentration of secondary and tertiary pond can satisfy 30mg/l secondary treatment standard. The SS concentration of effluent from tertiary pond, however, is slightly greater than the standard, which results from activities of carps growing in the pond. The average yearly total nitrogen concentration of influent is 206.8mg/l and that of the effluent from each pond is 48.6, 30.8, and 21.0mg/l respectively. Approximate 74, 88, and 90% of total nitrogen are removed in each pond. The mean yearly total phosphorous concentration of influent is 20.7mg/l and that of the effluent from each pond is 5.3, 3.2, and 2.1mg/l respectively. Approximate 97, 98, and 99% of total phosphorous are removed in each pond. The high removal of nitrogen and phosphorous results from active growth of algae in the upper layer of pond water. Important pond design parameters for southern part of Korea -- areal loading of BOD5, liquid depth, hydraulic detention time, free board, and pond arrangement -- are taken up.
To understand the relationship between water velocity in a fishway and fish morphology, the fineness ratio of fish, expressed as the standard length to the body depth, was measured. The fineness ratios of fish groups belonging to Cyprinidae, Acheilognathinae, Centropomidae, and Centrachidae were relatively low. The FRs of other groups, excluding eel-shaped types of fish, were over 4.5, indicating streamlined structures. The fineness ratios of Korean fish were classified into three different types: type I was Bitterling-Carp group that favored a slow-flowing pool habitat(FRs ranged from 2.1 to 3.3), type II was the Chub group representing streamlined types (FRs FRs ranged from 3.7 to 5.2), and type III was the Smelt-Barbel fish group that preferred riffle-run habitats with high velocity (FRs is over 5.2). Fish abundance analysis of fish using the fishway during the experimental periods showed a relatively high abundance of both type II and III compared to type I. The FRs of the fish passing through fishway(velocity 0.5-1.0 m s-1) ranged from 4.5 to 5.0, indicating that the fish using the fishway were mainly the streamlined type. As one of the standard fishways in Korea, the flow rate of the ice harbor type ranged from 0.2 to 2.6 m s-1. The FR values of the fish groups using the fishway ranged from 4.3 to 5.0. In contrast, the flow rate measured in an artificial channel type of fishway (same as a natural type of fishway) ranged from 0.1 to 1.9 m s-1 and the FR values for the fish groups using a natural type of fishway ranged from 3.3 to 5.3. The low FR values in natural fish are considered to be due to differences in the flow rates between the two types of fishways.
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