Kim, I.-D.;Ahn, M.-H.;Hur, T.-Y.;Hong, M.-P.;Seok, H.-B.
Journal of Embryo Transfer
/
v.19
no.2
/
pp.155-163
/
2004
The aims of this study are 1) to test oocytes and embryos collected from in-vitro to achieving the valuable protocol by culturing, vitrifying and thawing of oocytes/embryos, and 2) to transfer them to recipient, and finally have resulted in pregnancies from recipient females after surgical or nonsurgical transfer. In vitro maturation and fertilization were performed according to Funahashi et al (1994). Glucose-free NCSU 23 supplemented with 5 mM sodium pyruvate, 0.5 mM sodium lactate and 4 mg/ml bovine serum albumin for 2 days at $39^{\circ}C$, and 10% fetal bovine serum albumin was added to the culture medium thereafter. Embryos were treated with 7.5 ${\mu}g/ml$ cytochalasin-B for 30 min, centrifuged at 13,000 rpm for 13 min and then exposed sequentially to an ethylene glycol(EG) vitrification solution, aspirated into OPS, and plunged/thawed into/from liquid nitrogen. In vivo embryos were surgically collected from three dornors after AI for control group. Forty-nine embryos were washed 3 times in mPBS + 10% FBS, followed treatments : cultured, centrifuged, vitrified, recovered and transferred to recipients as in vitro prepared embryos. Three recipients were transferred individually with 100, 100 frozen embryos derived from abattoir and 34 fresh embryos by surgically, and another three recipients were transferred individually with 150, 150 frozen embryos and 100 fresh embryos by nonsurgically, respectively. all recipient sows exhibited delayed returns to estrus. To our knowledge, theses results suggest that required an improved techniques, more vigorous embryos preparation and substitute to gilt with cleaner uterous condition.
Rainfall factor. R, and soil factor, K were estimated to use the Revised Universal Soil Loss Equation (RUSLE) to predict the amount of soil erosion from a land on slope in Kangwon-do, Korea. The average of R factor was 405 with a range from 251 to 601. The R factor differed among regions. The R factor at Taegwalryung, in the highland region, was 409 and those at Inje and Hongchon, in the mid mountainous regions, ranged from 310 to 493. The R factors at Wonju and Chuncheon, in the plain regions, ranged from 505 to 601. The R factors at Sokcho, Kangnung and Samchok, in the east coastal region, which ranged from 251 to 368, were lowee than those in the western part of the Taebaeg Mountains. The R factor during the winter including the effect of winter freezing and thawing was 12 to 30% of the annual average value in the east coastal and highland regions, while that in the western part of Taebaeg Mountains was lower than 7%. The average of K factor in the surface soil was 0.21 with a range from 0.06 to 0.42. The K factors of Odae and Weoljeong serieses were the lowest, while that of Imog was the highest. The average of K factor in the subsoil was 0.28 with a range from 0.07 to 0.45. The K factor of the subsoil was 1.3 times higher than that of top soil. The average of K factor in he soil including the effect of the gravel covering and percolation was 0.18 with a range from 0.03 to 0.33. In contrast. the K factor excluding the effect of the gravel covering was lower than this. The average of K factor in the frozen subsoil was 0.33, which was 1.6 times higher than that of the non frozen subsoil.
The purpose of this research is to complement the existing researches on landfill bottom liners behavior during the periods of freeze and thaw. Landfill-related researches have been typically focused on small-scale soil samples that are often compacted under conditions different from those used in the field. Although these tests have been invaluable in clarifying the problem of freeze and thaw, extending the results of such experimental studies to prototype landfills are questionable. In this investigation, the author utilized a large scale laboratory simulation allowing inclusion of the field depth of the cover systems, layered soil profiles, rainfall simulation, a cold climate and boundary conditions similar to those encountered in the landfill. The soil materials were stabilized soils (mixed clays, cements, and minerals) instead of clays. The bottom liners are made up of drainage layer (30 cm), stabilized layer (75 cm), and leach collection layer (60 cm). The stabilized layers are made up of supporting layer (45 cm) and low permeable layer (30 cm) - consisting of $P_A\; and\; P_B$ layer. As a results, depths of penetration increased by about 2~5 more centimeters at rainfall simulated designs than those at no rainfall simulated designs (that is design 3, design 5 and design 7) - it increased by about 20mm/day in the bottom liners and frost heaves also increased it by a few millimeters. Also, a few cracks appeared partly. According to these results, we can surmise that the compacted stabilized soil is more reliable than the compacted clay liners for construction of the landfill liners.
This experiment was carried out to evaluate the sanitary quality of commercially frozen sea foods. One hundred and sixteen samples in six different items from several refrigeration plant in Busan city were examined from March to December in 1974. In addition, the changes in bacterial density through the process from thawing, round or semifilleted frozen alaska pollack to the finishing as frozen fillet blocks were observed. To evaluate the sanitary quality, sanitary indicative bacteria such as total coliform, fecal coliform, fecal streptococci and enterococci as well as plate counts were determined. From the results, the median value of fecal coliform MPN was 20 per 100 grams of the samples and that of enterococci was 790. The median value of plate counts was $2.2\times10^4$ per gram. The plate counts were not correlated with the number of sanitary indicative bacteria. The results suggest that enterococci could be used advantageously in preference to coliform organisms as indicative bacteria for the evaluation of sanitary quality of frozen sea foods. The plate counts at $20^{\circ}C$ of the samples were 14 times higher than that at $35^{\circ}C$. Geometric mean of total coliform MPN was 310 and that of enterococci was 143. Bacterial density was reduced by fleering. Morethan 50 percent for total coliform MPN and $35^{\circ}C$ plate counts, and about 35 percent for enterococci MPN and $20^{\circ}C$ plate counts were reduced under the contact freezing unit which was generally operated at $-40^{\circ}C$. About fifty-five percent of the samples were negative in fecal coliform test and 10 percent of those were exceeded $1.0\times10^5$ per gram in $35^{\circ}C$ plate counts.
Cryopreservation induces sublethal damage to the spermatozoa, which leads to their reduced fertile life. This study was designed to determine effect of glycerol and ethylene glycol as cryoprotectant in extender on improve the freezability of Jeju horse semen. The semen was cryopreserved with glucose-EDTA extender containing each 5% glycerol, 5% ethylene glycol, 8% glycerol or 8% ethylene glycol, respectively. Post-thawed sperm were evaluated motility, viability, Membrane integrity and acrosome integrity. Post-thawed sperm motility were not significantly differences among treatments. However, sperm viability were significantly higher (p<0.05) in 8% glycerol ($39.85%{\pm}11.41$) than in 5% glycerol treatment ($18.08%{\pm}1.61$). In membrane integrity, swelling sperm ratio was significantly higher (p<0.05) in 8% glycerol ($34.12%{\pm}11.02$) than other treatments. In the percentage of capacitated sperm assessed by CTC staining, F pattern was significantly higher in 8% ethylene glycol than 5% glycerol and 5% ethylene glycol (p<0.05). B pattern ratio was significantly increased in 5% ethylene glycol compared with 8% glcerol and 8% ethylene glycol (p<0.05). Moreover, 8% ethylene glycol treatment was significantly decreased AR pattern ratio compared with other treatments (p<0.05). It is concluded that treatment of 8% glycerol was improved the sperm viability and 8% ethylene glycol was improved the sperm ascrosome integrity after thawing. However, they were not significantly difference between 8% glycerol and 8% ethylene glycol on post-thawed sperm viability. Therefore, 8% ethylene glycol was more effective sperm cryoprotectant than 8% glycerol in Jeju Horse.
Kim, M.K.;Baik, C.S.;Uhm, S.J.;Kim, E.Y.;Yoon, S.H.;Park, S.P.;Chung, K.S.;Lim, J.H.
Clinical and Experimental Reproductive Medicine
/
v.23
no.3
/
pp.379-384
/
1996
This study was undertaken to investigate the sister chromatid exchange (SCE) frequency and embryonic development after exposure to cryoprotectants and vitrification of mouse zygotes. Mouse IVF zygotes were cryopreserved by vitrification using vitrification solution, EFS40 (40% ethylene glycol, 30% Ficoll and 0.3 M sucrose in phosphate buffer saline containing 10% FBS). After mouse zygotes were exposed to EFS40 at $25^{\circ}C$ for 30 sec., they were immediately plunged into $LN_2$ or cultured for cryoprotectant toxicity test without freezing. The results obtained in these experiments were summarized as follows: After thawing, survival rates to the 2-cell stage of zygotes exposed to or vitrified in EFS 40 (98.5%, 95.2%) were not significantly difference compared with that of control (100%). However, the developmental rates upto blastocyst and hatching blastocyst in vitrified groups (66.7, 50.0%) were lower than those of control (93.9, 81.8%) or exposed group (94.0, 78.8%) (p<0.05). When the influence of vitrification and exposure to cryoprotectant on the in vitro development of mouse zygotes was assessed by the SCE frequency, the SCE frequency in exposed ($20.2{\pm}2.1$) to or vitrified embryos ($21.4{\pm}3.2$) was higher than that in control embryos ($16.8{\pm}1.5$). These results suggest that the frequency of SCE was increased after cryoprotectant exposure or Vitrification although developmental rates of zygotes upto blastocysts and /or hatching blastocysts were not afected by cryoprotectant.
Global warming has made the polar regions more accessible, leading to increased demand for the construction of new resource-development plants in oil-rich permafrost regions. The selection of locations of resource-development plants in permafrost regions should consider the surface displacement resulting from thawing and freezing of the active layer of permafrost. However, few studies have considered surface displacement in the selection of optimal locations of resource-development plants in permafrost region. In this study, Analytic Hierarchy Process (AHP) analysis using a range of geospatial information variables was performed to select optimal locations for the construction of oil-sands development plants in the permafrost region of southern Athabasca, Alberta, Canada, including consideration of surface displacement. The surface displacement velocity was estimated by applying the Small BAseline Subset Interferometric Synthetic Aperture Radar technique to time-series Advanced Land Observing Satellite Phased Array L-band Synthetic Aperture Radar images acquired from February 2007 to March 2011. ERA5 reanalysis data were used to generate geospatial data for air temperature, surface temperature, and soil temperature averaged for the period 2000~2010. Geospatial data for roads and railways provided by Statistics Canada and land cover maps distributed by the North American Commission for Environmental Cooperation were also used in the AHP analysis. The suitability of sites analyzed using land cover, surface displacement, and road accessibility as the three most important geospatial factors was validated using the locations of oil-sand plants built since 2010. The sensitivity of surface displacement to the determination of location suitability was found to be very high. We confirm that surface displacement should be considered in the selection of optimal locations for the construction of new resource-development plants in permafrost regions.
The physiological and biochemical role of potassium for upland crops according to recent research reports and the nutritional status of potassium in Korea were reviewed. Since physical and chemical characteristics of potassium ion are different from those of sodium, potassium can not completely be replaced by sodium and replacement must be limited to minimum possible functional area. Specific roles of potassium seem to keep fine structure of biological membranes such as thylacoid membrane of chloroplast in the most efficient form and to be allosteric effector and conformation controller of various enzymes principally in carbohydrate and protein metabolism. Potassium is essential to improve the efficiency of phoro- and oxidative- phosphorylation and involve deeply in all energy required metabolisms especially synthesis of organic matter and their translocation. Potassium has many important, physiological functions such as maintenance of osmotic pressure and optimum hydration of cell colloids, consequently uptake and translocation of water resulting in higher water use efficiency and of better subcellular environment for various physiological and biochemical activities. Potassium affects uptake and translocation of mineral nutrients and quality of products. potassium itself in products may become a quality criteria due to potassium essentiality for human beings. Potassium uptake is greatly decreased by low temperature and controlled by unknown feed back mechanism of potassium in plants. Thus the luxury absorption should be reconsidered. Total potassium content of upland soil in Korea is about 3% but the exchangeable one is about 0.3 me/100g soil. All upland crops require much potassium probably due to freezing and cold weather and also due to wet damage and drought caused by uneven rainfall pattern. In barley, potassium should be high at just before freezing and just after thawing and move into grain from heading for higher yield. Use efficiency of potassium was 27% for barley and 58% in old uplands, 46% in newly opened hilly lands for soybean. Soybean plant showed potassium deficiency symptom in various fields especially in newly opened hilly lands. Potassium criteria for normal growth appear 2% $K_2O$ and 1.0 K/(Ca+Mg) (content ratio) at flower bud initiation stage for soybean. Potassium requirement in plant was high in carrot, egg plant, chinese cabbage, red pepper, raddish and tomato. Potassium content in leaves was significantly correlated with yield in chinese cabbage. Sweet potato. greatly absorbed potassium subsequently affected potassium nutrition of the following crop. In the case of potassium deficiency, root showed the greatest difference in potassium content from that of normal indicating that deficiency damages root first. Potatoes and corn showed much higher potassium content in comparison with calcium and magnesium. Forage crops from ranges showed relatively high potassium content which was significantly and positively correlated with nitrogen, phosphorus and calcium content. Percentage of orchards (apple, pear, peach, grape, and orange) insufficient in potassium ranged from 16 to 25. The leaves and soils from the good apple and pear orchards showed higher potassium content than those from the poor ones. Critical ratio of $K_2O/(CaO+MgO)$ in mulberry leaves to escape from winter death of branch tip was 0.95. In the multiple croping system, exchangeable potassium in soils after one crop was affected by the previous crops and potassium uptake seemed to be related with soil organic matter providing soil moisture and aeration. Thus, the long term and quantitative investigation of various forms of potassium including total one are needed in relation to soil, weather and croping system. Potassium uptake and efficiency may be increased by topdressing, deep placement, slow-releasing or granular fertilizer application with the consideration of rainfall pattern. In all researches for nutritional explanation including potassium of crop yield reasonable and practicable nutritional indices will most easily be obtained through multifactor analysis.
In a basic attempt to develop the prophylactic and therapeutic measures on intestinal giantcystic disease of the Israel carp, C), prinks carpio nudum, the effects of physical and chemical factors on viability or survival of the spores of Thelchcnellus kiteuei were checked in vitro by means of extrusion test on the polar filament. When the fresh spores suspended with 0.45% and 0.9% scdium chloride solution and distilled water were laid at $5^{\circ}C$ and $28^{\circ}C$ for short terms, the extrusion rates increased until the 3rd day, meanwhile when son;e of them were suspended with Tyrode's solution at $-70^{\circ}C$ the rates increased gradually until the 8th day. Viabilities of the spores suspended with 0.9% saline and added antibiotics to the suspension at $5^{\circ}C$ for long terms lasted for 997 days and 1, 256 days (presumed values) at maximum, respectively. The spores suspended with distilled water at $28^{\circ}C$ for long terms survived 152.4 days, but the spores suspended with Tyrode's solution at $-70^{\circ}C$ for long terms showed almost the same viable pattern as early freezing stages up to 780 days. The spores suspended with Tyrode's solution, frozen at $-70^{\circ}C$ and thawed at $5^{\circ}C$, showed the highest rate of extrusion of the polar filament. In the case of frozen spores, the extrusion rates during heating tend to become higher in accordance with the increase of frozen period, and the critical points of 180 day-frozen spores to be killed were generally 78.5 hr. at $60^{\circ}C$, 23.4 hr. at $70^{\circ}C$, 189.1 min. at $80^{\circ}C$ or 10.5 min. at $90^{\circ}C$. The longer the spores were frozen, the more time was needed for the death of spores after thawing; 20 days-17.4 days, 100 days-33.2 days, and 400 days-37.8 days. The longer the spores were frozen, the more time was needed for the death of spores at a conventional when they were dried air drying condition, 540 days-23.5 days, 160 days-21.0 days, and 20 days-14.4 days. On the other hand, the longer the spores were frozen, the more spores were dead rapidly when they were irradiated with 10W UV-ray; 100 days-26.0 hr, 300 days-21.9 hr, and 540 days-13.9 hr. The time needed for killing 200 days-frozen spores by various disinfectants at 1, 000 ppd was 5.2 min. by calcium oxide, 10.4 min. by potassium permanganate, 27.8 min. by malachite green and 14.3 hr. by formalin. Transient inhibitory effects of the extrusion of the polar filament were observed by various antiprotozoal and antifungal agents in the descending order of ketoconazole. metronidasole and dapsone. The above results presume that full drying, followed by spraying CaO and maintaining sunny condition for a few days on the concrete bottoms of knish farm may be an effective method for the prevention of intestinal giant.cystic disease.
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