• Title/Summary/Keyword: High RPM

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The effects GeongshinhaeGihwan 1(GGT1) has on the hGHTg (human growth hormone transgenic) obese male rats' blood-antiobestic index (형질전환 비만모델 수컷 hGHTg rats에서 경신해지환(輕身解脂丸)이 혈중 항비만지표에 미치는 영향)

  • Jung, Yang-Sam;Tsung, Pei-Chin;Choi, Seung-Bae;Kim, Gyeong -Cheol;Shin, Soon-Shik
    • Herbal Formula Science
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    • v.13 no.2
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    • pp.1-16
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    • 2005
  • Objectives: To find out the effects GGTl, an antiobestic drug widely used in clinics, has on the blood-antiobestic index and the toxicity index using the data from the hGHTg obese male rats. We looked closely into both of the two indices because GGTl antiobestic effect can happen not only by pharmacological action, but also by its toxicity. Also, we verified the difference in effect between GGTl and reductil (sibutramine), which has been approved by the FDA of the United States. Methods: After performing the experiments for 8 weeks on the hGHTg obese male rats divided into three groups: the control group, the GGTl group, and the reductil (RD) group, we anesthetized the rats with Diethyl ether and took a 3ml blood sample from the heart. Then, after coagulating the blood in room temperature by using the plasma separator, we centrifuged it for 25 minutes in 3,000rpm using the high-speed refrigerated centrifuge. We kept the separated plasma in a deep freezer at $-80^{\circ}C$, and repeatedly measured the antiobestic index and the toxicity index twice using the hematology biochemistry analyzer. Also, in order to judge the indirect toxicity index, we separated liver from kidney and observed them. Results: When we looked at the results of the analysis of covariance on the measuring elements related to the antiobestic index (TC, HDL, LDL, TG, and GLU), there was no significant difference among the groups in all measuring elements. Also, the results of the analysis of covariance on the two roups (RD group and GGTl group) showed that the p-values had no significant difference under the level of significance 0.05. When we looked at the result of the analysis of covariance on the measuring elements related to the toxicity index (GOT, GPT, GGT, CREA, UA, ALB, and TP), we could see that the p-values in GPT, ALB, and TP have a significant difference among the groups. Also, the results of the analysis of covariance about the measuring elements related to the toxicity index on both groups, RD group and GGTl group, showed no significant difference in the p-values of all of the measuring elements in the two groups, RD and GGTl group. Conclusions: In conclusion, through this experiment, the safety of GGTl has been approved, and although the verification on its medical effect has not been clearly approved, when we consider the fact that it belongs to the same group as reductil, an antiobestic drug approved by the FDA of the United States, we could indirectly verify that GGTl has an antiobestic effect. We believe that when doing a sample design for a future experiment, it needs to be performed on a greater sample size based on the power analysis that needs to be performed primarily in experiments, and a more accurate verification is needed through more systematic experiment plans.

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Characterization of Pretreatment for Barley straw by Alkaline Solutions (염기 용매를 이용한 보릿짚의 전처리 특성)

  • Kim, Kyoung-Seob;Kim, Jun Seok
    • Korean Chemical Engineering Research
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    • v.50 no.1
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    • pp.18-24
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    • 2012
  • Lignocellulose is difficult to hydrolyze due to the presence of lignin and the technology developed for cellulose fermentation to ethanol is not yet economically viable. However, recent advances in the extremely new field of biotechnology for the ethanol production are making it possible to use of Agriculture residual biomass, e.q., Barley straw, because of their several superior aspects as Agriculture residual biomass; low lignin, high contents of carbohydrates. Barley straw consists of 39.78% cellulose (glucose), 22.56% hemicelluloses and 19.27% lignin. Pretreatment of barley straw using NaOH pretreatment solutions concentration with 2%, temperature $85^{\circ}C$ and reaction times 1 hr were investigates. $NH_4OH$ pretreatment condition was solutions concentration with 15%, temperature $60^{\circ}C$, and reaction times 24hr were investigates. Furthermore, enzymatic saccharification using cellulose at $50^{\circ}C$, pH 4.8, 180 rpm for conversion of cellulose contained in barley straw to monomeric sugar. The pretreatment of barley straw using NaOH and $NH_4OH$ can significantly improve enzymatic saccharification of barley straw by extract more lignin and increasing its accessibility to hydrolytic enzymes. The result showed NaOH pretreatment extracted yield of lignin was 24.15%. $NH_4OH$ pretreatment extracted yield of lignin was 29.09%. Shaccharification of barley straw pretreatment by NaOH for 72hr and pH 4.8 result in maximum glucose concentration 15.39g/L (58.40%) and by $NH_4OH$ for 72hr and pH 4.8 result in maximum glucose concentration 16.01g/L (64.78%).

Improving Corsican pine somatic embryo maturation: comparison of somatic and zygotic embryo morphology and germination

  • Wtpsk, Senarath;Shaw, D.S.;Lee, Kui-Jae;Lee, Wang-Hyu
    • Proceedings of the Plant Resources Society of Korea Conference
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    • 2003.04a
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    • pp.61-62
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    • 2003
  • Clonal propagation of high-value forest trees through somatic embryogenesis (SE) has the potential to rapidly capture the benefits of breeding or genetic engineering programs and to improve raw material uniformity and quality. A major barrier to the commercialization of this technology is the low quality of the resulting embryos. Several factors limit commercialization of SE for Corsican pine, including low initiation rates, low culture survival, culture decline causing low or no embryo production, and inability of somatic embryos to fully mature, resulting in low germination and reduced vigour of somatic seedlings. The objective was to develop a Corsican pine maturation medium that would produce cotyledonary embryos capable of germination. Treatments were arranged in a completely randomized design. Data were analyzed by analysis of variance, and significant differences between treatments determined by multiple range test at P=0.05. Corsican pine (Pinus nigra var. maritima) cultures were initiated on modified !P6 medium. Modifications of the same media were used for culture multiplication and maintenance. Embryogenic cultures were maintained on the same medium semi solidified with 2.5 g/l Gelrite. A maturation medium, capable of promoting the development of Corsican pine somatic embryos that can germinate, is a combination of iP6 modified salts, 2% maltose, 13% polyethylene glycol (PEG), 5 mg!l abscisic acid (ABA), and 2.5 g/l Gelrite. After initiation and once enough tissue developed they were grown in liquid medium. Embryogenic cell suspensions were established by adding 0.951.05 g of 10- to 14-day-old semisolid-grown embryogenic tissue to 9 ml of liquid maintenance media in a 250ml Erlenmeyer flask. Cultures were then incubated in the dark at 2022$^{\circ}$C and rotated at 120 rpm. After 2.53 months on maturation medium, somatic embryos were selected that exhibited normal embryo shape. Ten embryos were placed horizontally on 20 ml of either germination medium ($\frac{2}{1}$strength Murashige and Skoog (1962) salts with 2.5 g/l activated charcoal) or same medium with copper sulphate adjusted to 0.25 mg/1 to compensate for copper adsorption by activated carbon. 2% and 4% maltose was substituted by 7.5% and 13% PEG respectively to improve the yield of the embryos. Substitution of' maltose with PEG was clearly beneficial to embryo development. When 2% of the maltose was replaced with 7.5% PEG, many embryos developed to large bullet-shaped embryos. At latter stages of development most embryos callused and stopped development. A few short, barrel-shaped cotyledonary embryos formed that were covered by callus on the sides and base. When 4% of the maltose was removed and substituted with 13% PEG, the embryos developed further, emerging from the callus and increasing yield slightly. Microscopic examination of the cultures showed differing morphologies, varying from mostly single cells or clumps to well-formed somatic embryos that resembled early zygotic embryos only liquid cultures with organized early-stag. A procedure for converting and acclimating germinants to growth in soil and greenhouse conditions is also tested. Seedling conversion and growth were highly related to the quality of the germinant at the time of planting. Germinants with larger shoots, longer, straighter hypocotyls and longer roots performed best. When mature zygotic embryos germinate the root emerges, before or coincident with the shoot. In contrast, somatic embryos germinate in reverse sequence, with the cotyledons greening first, then shoot emergence and then, much later, if at all, the appearance of the root. Somatic seedlings, produced from the maturation medium, showed 100% survival when planted in a field setting. Somatic seedlings showed normal yearly growth relative to standard seedlings from natural seed.

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Comparison of Antibacterial and Antioxidant Activities of Mushroom Mycelium Culture Extracts Cultivated in the Citrus Extracts (감귤농축액 첨가배지에서 배양한 버섯균사체 추출물의 항균활성 및 항산화활성 비교)

  • Kim Man-Chul;Kim Min-Joo;Kim Taeg;Park Guen-Tae;Son Hong-Joo;Kim Gi-Young;Choi Woo-Bong;Oh Duck-Chul;Heo Moon-Soo
    • KSBB Journal
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    • v.21 no.1 s.96
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    • pp.72-78
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    • 2006
  • This study was carried out to investigate the antimicrobial and antioxidative effects of mycelium cultural extract from mushroom. Mushroom mycelium was grown in a defined synthetic liquid medium and citrus extracts, and the culture extracts were examined for antioxidant and antibacterial activity. Myceliums of Phellinus linteus, Cordyceps militaris, Coriolus versicolor, Sparassic crispa, Agaricus blazei, lnonotus obliquus, Lentinus edodes, Hericium erinacium, Gonoderma lucidium in 10% citrus extract supplemented medium and synthesis medium were incubated in a shaking incubator (120 rpm, $24{\sim}30^{\circ}C$ ) for $7{\sim}15$ days. The antimicrobial activity of the culture fluid of mushroom mycelium grown in submerged liquid culture was tested against 12 microorganisms which were fish pathogens and common bacterial species. The culture extracts showed high activity against Vibrio sp. and had poor effect on Streptocouus sp., S. parauberis, S. iniae. The culture extracts obtained from the synthetic medium showed $30{\sim}93%$ of the 1,1-diphenyl-2-picrylhydrazyl radical scavenger activity, the culture extracts obtained from the citrus extracts medium exhibited antioxidant activity up to 55%.

Production Medium Optimization for Monascus Biomass Containing High Content of Monacolin-K by Using Soybean Flour Substrates (기능성 원료를 기질로 이용하는 Monacolin-K 고함유 모나스커스 균주의 생산배지 최적화)

  • Lee, Sun-Kyu;Chun, Gie-Taek;Jeong, Yong-Seob
    • KSBB Journal
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    • v.23 no.6
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    • pp.463-469
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    • 2008
  • During the last decade, monacolin-K biosynthesized by fermentation of red yeast rice (Monascus strains) was proved to have an efficient cholesterol lowering capability, leading to rapid increase in the market demand for the functional red yeast rice. In this study, the production medium composition and components were optimized on a shake flask scale for monacolin-K production by Monascus pilosus (KCCM 60160). The effect of three different soybean flours on the monacolin-K production were studied in order to replace the nitrogen sources of basic production medium (yeast extract, malt extract and beef extract). Among the several experiments, the production medium with dietary soybean flour to replace a half of yeast extract was very good for monacolin-K production. Plackett-Burman experimental design was used to determine the key factors which are critical to produce the biological products in the fermentation. According to the result of Plackett-Burman experimental design, a second order response surface design was applied using yeast extract, beef extract and $(NH_4)_2SO_4$ as factors. Applying this model, the optimum concentration of the three variables was obtained. The maximum monacolin-K production (369.6 mg/L) predicted by model agrees well with the experimental value (418 mg/L) obtained from the experimental verification at the optimal medium. The yield of monacolin-K was increased by 67% as compared to that obtained with basic production medium in shake flasks.

Method Development for Determination of Chlorogenic Acid and Arbutin Contents in Fruits by UHPLC-MS/MS (UHPLC-MS/MS를 이용한 과일류 중 클로로젠산 및 알부틴 동시분석법 개발)

  • Choi, Young-Ju;Jeon, Jong-Sup;Kim, Woon-Ho;Jung, You-Jung;Ryu, Ji-Eun;Choi, Jong-Chul;Chae, Kyung-Suk;Lee, Jin-Hee;Do, Young-Sook;Park, Young-Bae;Yoon, Mi-Hye
    • Journal of Food Hygiene and Safety
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    • v.34 no.5
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    • pp.413-420
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    • 2019
  • In this study, a sample preparation method and a simultaneous determination method by ultra-high performance liquid chromatography coupled with tandem mass spectrometry for 9 isomers of chlorogenic acid and arbutin in fruits were developed. The samples were extracted using 90% methanol (pH 3.0), with the solutions being shaken and then sonicated for 10 min each. After centrifugation at 4,000 rpm for 10 min, the extraction was concentrated under a vacuum at $40^{\circ}C$ using a vacuum evaporator. The residue was dissolved in 5 mL of 5% methanol and filtered through a $0.45{\mu}m$ membrane before UHPLC-MS/MS analysis. The separations were performed on a C18 column with gradient elution of water (containing 0.1% formic acid) and methanol (containing 0.1% formic acid). The specificity, linearity, limit of detection, limit of quantification, accuracy, and precision of the proposed methods were also evaluated.

Quantitative Elemental Analysis in Soils by using Laser Induced Breakdown Spectroscopy(LIBS) (레이저유도붕괴분광법을 활용한 토양의 정량분석)

  • Zhang, Yong-Seon;Lee, Gye-Jun;Lee, Jeong-Tae;Hwang, Seon-Woong;Jin, Yong-Ik;Park, Chan-Won;Moon, Yong-Hee
    • Korean Journal of Soil Science and Fertilizer
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    • v.42 no.5
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    • pp.399-407
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    • 2009
  • Laser induced breakdown spectroscopy(LIBS) is an simple analysis method for directly quantifying many kinds of soil micro-elements on site using a small size of laser without pre-treatment at any property of materials(solid, liquid and gas). The purpose of this study were to find an optimum condition of the LIBS measurement including wavelengths for quantifying soil elements, to relate spectral properties to the concentration of soil elements using LIBS as a simultaneous un-breakdown quantitative analysis technology, which can be applied for the safety assessment of agricultural products and precision agriculture, and to compare the results with a standardized chemical analysis method. Soil samples classified as fine-silty, mixed, thermic Typic Hapludalf(Memphis series) from grassland and uplands in Tennessee, USA were collected, crushed, and prepared for further analysis or LIBS measurement. The samples were measured using LIBS ranged from 200 to 600 nm(0.03 nm interval) with a Nd:YAG laser at 532 nm, with a beam energy of 25 mJ per pulse, a pulse width of 5 ns, and a repetition rate of 10 Hz. The optimum wavelength(${\lambda}nm$) of LIBS for estimating soil and plant elements were 308.2 nm for Al, 428.3 nm for Ca, 247.8 nm for T-C, 438.3 nm for Fe, 766.5 nm for K, 85.2 nm for Mg, 330.2 nm for Na, 213.6 nm for P, 180.7 nm for S, 288.2 nm for Si, and 351.9 nm for Ti, respectively. Coefficients of determination($r^2$) of calibration curve using standard reference soil samples for each element from LIBS measurement were ranged from 0.863 to 0.977. In comparison with ICP-AES(Inductively coupled plasma atomic emission spectroscopy) measurement, measurement error in terms of relative standard error were calculated. Silicon dioxide(SiO2) concentration estimated from two methods showed good agreement with -3.5% of relative standard error. The relative standard errors for the other elements were high. It implies that the prediction accuracy is low which might be caused by matrix effect such as particle size and constituent of soils. It is necessary to enhance the measurement and prediction accuracy of LIBS by improving pretreatment process, standard reference soil samples, and measurement method for a reliable quantification method.

The Studies on the Physiological Active Substances of Mugwort Components for the Utilization to the Foods of Animal Husbandry (축산식품에 이용하기 위한 쑥 성분중의 생리활성에 관한 연구)

  • Lee, Chi-Ho
    • Proceedings of the Korean Society for Food Science of Animal Resources Conference
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    • 1998.05a
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    • pp.37-54
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    • 1998
  • This study was conducted to investigate the effects of mugwort extracts on the blood ethanol concentration, liver function and low level of cadmuim(Cd) in rats. The effects of mugwort extracts on the blood ethanol concentration was studied in Sprague-Dawley rats (10 weeks old) administered p.o. with 25% ethanol (5g/1kg body weight) and then injected with mugwort extracts (at the 2% levels of daily feed consumption compared with the concentration of catechins level in mugwort extracts) in caudal vein. SD rats were divided into five groups : control group (CON-E, only ethanol and 0.85% saline sol'n treated instead of each extracts), water extracts of mugwort treated to the control (MDW-E), ethanol extracts of mugwort treated to the control (POH-E). And then rat plasma of each time (0hr, 1hr, 2hr, 3hr) was investigated ethanol concentration by gas chromatography. Another rats were measured at the time of 0 and 5hr for the test of GOD(Glutamic Oxaloacetic Transaminase) and GPT(Glutamic Pyruvic Transaminase). Components of each extracts were analyzed by using high performance liquid chromatography. The effects of mugwort extracts on the liver function were studied in culture of rat hepatocyte composed of three groups : Control group and two groups treated with each extracts (1% & 2% MDW, 1% & 2% MOH). Condition of rat hepatocytes cultured for 36hr at $37^{\circ}C$(5% $CO_2$ incubator), number of cells, GOT and GPT activity were investigated. The results obtained were summarized as follows ; 1. Catechins level of mugwort extracts was $8{\sim}10mg/100g(MDW)$, $3{\sim}4mg/100g(MOH)$ 2. The contents of (-)-Epigallocatechin was high in MDW 3. The effects of mugwort extracts on the blood ethanol concentration were as follows; 1) The order in ethanol degradation efficiency was MDW-E > MOH-E > CON-E. 2) Ethanol concentration significantly decreased (p<0.05) in MDW-E and MOH-E. 4. The effects of mugwort extracts on the liver function were as follows; (rat hepatocytes cultured for 36hr at $37^{\circ}C$) 1) Cells condition of MDW-L was better than other groups. 2) The order in number of cells (rat hepatocytes) was 2% MDW-L >1% MDW-L >1% MOH-L > Con-L > 2% MOH-L 5. Cd treatment increased concentrations of hepatic GSH level, and decreased GOT activity in plasma. Therefore, this results suggest that the effects of mugwort extracts may an important rols in degradation ethanol and recovery liver function in body. Also, Mugwort extracts may modify the toxicities of Cd in Cd-treated rats and play an important roles in preventing the liver from various toxicants including Cd in Cd treated rats.

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An Analysis of the Imported Consumer Goods Distribution Sector of Korea: From a Vertical Structure Viewpoint (수입소비재(輸入消費財) 유통구조(流通構造)의 효율화(效率化) 방안(方案))

  • Nam, Il-chong
    • KDI Journal of Economic Policy
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    • v.13 no.1
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    • pp.3-33
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    • 1991
  • Since the early 1980's, the Korean government has gradually been widening the Korean market to foreign consumer goods. This, combined with the increased purchasing power of the Korean consumers resulting from the continued economic growth of the country, has sparked a spectacular influx of foreign consumer goods into Korea, ranging from BMW's to chopsticks. Import of foreign consumer goods amounted to more than 6 billion dollars in 1989 and is continuing to grow at a rapid rate. The increased import of foreign consumer goods doubtlessly improved the overall welfare of the Korean consumers by providing them with a wider range of options to choose from, by lowering the prices of some of the consumer goods domestically produced, and also by forcing the producers of some Korean goods to face competition with better foreign goods, thus giving them an incentive to raise the quality of their products. However, it is agreed by most economists that this increase in general welfare has been much smaller than what they had expected at the outset. Consumer prices of most imported consumer goods are easily double the import price, and in some cases, more than treble the import prices. Further, there has not been a noticeable drop in the prices of domestically produced consumer goods. Much of the blame has been attributed to the distribution sector of Korea. The objective of this paper is to analyze the imported consumer goods distribution sector of Korea, focusing on the possible sources of the poor performance of that sector, and to make policy suggestions that could potentially increase the welfare. This paper differs from all the previous research by others on this subject in that it analyzes the imported consumer goods distribution sector of Korea as a vertical structure. The distribution sector of an imported consumer good is a vertical structure since it consists of an international market, an import stage, and domestic wholesale and retail markets, in that order vertically. Our study naturally includes the analysis of the vertical restraints as well as the analysis of the industrial organization of each horizontal stage in the vertical structure. Each horizontal component of the imported consumer goods distribution sector is basically a monopolistically competitive market differentiated by characteristics of goods and by the locations and the services of firms. Further, restrictive dealership and resale price maintenance are found to be widely in use. Our main findings are the follwing; First, most consumer goods are imported monopolistically or oligopolistically through restrictive dealership contracts between foreign producers and domestic importers. Such restrictive dealership gives importers market power in the domestic market and explains many of the large discrepancies betwen the consumer prices and the import prices of many goods. Korean anti - trust law does not cover the issues arising from the market power of an importer resulting from a restrictive dealership contract. Second, some major producers of Korean goods are also importers of foreign goods that are substitutes of their products. The import of substitutes by major domestic producers is anti - competitive because it tends to raise the prices of both domestic goods and foreign goods, and also because it reduces the incentive of the domestic producers to raise the quality of their products. Third, wholesalers and retailers widely use resale price maintenance as a price fixing mechanism, and while this is against the anti- trust law, it seldom gets noticed. Fourth, the high level of rents of real estate for commercial use works as an entry barrier to the distribution sector and results in reduced competition by the firms in that sector. Finally, there are information problems. Consumers have inferior information to firms about the quality of a foreign consumer good that they have not tried before. Such information asymmetry often enables firms to raise prices. In addition, information asymmetry between importers frequently delays the import of cheaper substitutes. In order to alleviate the problems indentified above, we suggest the following policy changes. The government should strengthen the anti - trust law and its enforcement to regulate restrictive import contracts, import of competing goods by major domestic producers, and RPM by wholesalers and retailers that is aimed at price fixing. In addition, the government should loosen its tight real estate policy to encourage investment in the distribution sector. Finally, we suggest that the import price revelation policy that has been in use for some items since 1990 be expanded to most imported consumer goods that are introduced for the first time to give consumer better information and be used only for the period of time needed to inform sufficient number of consumers.

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Studies on Genetics and Breeding in Rainbow Trout(Oncorhynchus mykiss) VII. Fertilization of Fresh Egg with Co-Preserved Sperm and Ultrastructural Changes (무지개 송어의 유전 육종학적 연구 VII. 동결보존시킨 정자와 신선한 난모세포의 수정 및 미세구조적 변화)

  • PARK Hong-Yang;YOON Jong-Man
    • Korean Journal of Fisheries and Aquatic Sciences
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    • v.25 no.2
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    • pp.79-92
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    • 1992
  • This study was carried out to develop new techniques useful for cryopreservation, thawing and artificial insemination, and ultrastructural changes of cryopreserved spermatozoa in rainbow trout(Oncorhynchus mykiss) . Two extenders, such as Tyrode solution and Whittingham's $T_6$ solution, were used to preserve rainbow trout sperm in refrigerator $(-20,\;-40\;and\;-70^{\circ}C)$ or liquid nitrogen $%(-196^{\circ})$. Hand-stripped semen was diluted to 1:16 with two extenders, an then the semen were frozen after mixing semen and each extender containing 1M or 1.5M DMSO solution to 1:1. After 60 days cryopreserved semen was thawed in a $13^{\circ}$ water bath, and subsequently centrifugated. After centrifugation at 1,000 rpm for 5 min thawed semen was washed with extenders, and then fertilized with fresh eggs. The results obtained in these experiments were summarized as follows: After cryopreservation, over 75% of spermatozoa were appeared motile and the survival rate was high. Following cryopreservation by the addition of cryoprotectant such as DMSO, methanol and glycerol, the fertilization rate of the thawed spermatozoa appeared over $99\%$ compared with the control having $99\%$ of fertilization rate. There was no difference between the control and experimental groups such as $(-20^{\circ}C\;-40^{\circ}C\;and\;-70^{\circ}C)$ and $-196^{\circ}$ in fertilization rate. Following cryopreservation at $-196^{\circ}$ by the addition of 1M DMSO of cryoprotectant, each fertilization rate following 24 hours and hatching rate following 24 days showed $96\%$ and $8\%$ by the addition of BSA, but showed $98\%\;and\;10%$ by no addition of BSA. Following 2 months of cryopreservation by the addition of 1M DMSO of cryoprotectant, there were $10%$ of hatching rate at $-196^{\circ}\;and\;10\%\;and\;35\%,\;respectively,\;at\;-40^{\circ}C\;and\;-70^{\circ}C$. Following 2 months of cryopreservation by the addition of 1M methanol of cryoprotectant, there were $22\%$ of fertilization rate at $-20^{\circ}C,\;and\;28\%,\;at\;-70^{\circ}C$ Following 2 months of cryopreservation by the addition of 1M glycerol of cryoprotectant, there were $22\%$ of fertilization rate at $-20^{\circ}C$, and $33\%,\;at\;-70^{\circ}C$. pollowing 2 months of cryopreservation by the addition of 1.5M DMSO of cryoprotectant, there were $27\%$ of fertilization rate at $-20^{\circ}C,\;an\;36\%\;and \;35\%,\;respectively,\;at\;-40^{\circ}C\;and\;-70^{\circ}C$. Following 2 months of cryopreservation by the addition of 1.5M glycerol of cryoprotectant, there were $34\% \;of\;fertilization\;rate\;at\;-20^{\circ}C, \;and\;31\%\;and\;31\%,\;respectively,\;at \;-40^{\circ}C\;and\;-70^{\circ}$. Following 2 months of cryopreservation by the addition of 1.5M methanol of cryoprotectant, there were $28\%$ of fertilization rate at $-20^{\circ}C,\;and\;29\%\;and\;28\%,\;respectively,\;at\;-40^{\circ}C\;and\;-70^{\circ}C.$ From 10 days and 15 days following fertilization at $13^{\circ}C\;and\;10^{\circ}C$, respectively, the mortality rate of fertilized ova was markedly increased. The middle piece of spermatozoa had two set of central doublets, nine set of outer coarse fibres, and mitochondrial sheath. Spermatozoa went through morphological changes during storage, e.g. winding of flagella, detachment of the nuclear envelope and the plasma membrane from the nucleus of the sperm head. There were $1\%$ abnormal spermatozoa in fresh sperm and about $15\%$ during storage.

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