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Detection of Salmonella spp. in Seafood via Desalinized DNA Extraction Method and Pre-culture (전배양과 탈염과정을 포함하는 DNA 추출법을 이용한 분자생물학적 방법으로 수산물 중 오염된 Salmonella spp.의 검출)

  • Ye-Jun Song;Kyung-Jin Cho;Eun-Ik Son;Du-Min Jo;Young-Mog Kim;Seul-Ki Park
    • Journal of Food Hygiene and Safety
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    • v.38 no.3
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    • pp.123-130
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    • 2023
  • Salmonella spp. are prevalent foodborne pathogens that are infective at relatively low concentrations, thus posing a serious health threat, especially to young children and the elderly. In several countries, the management and regulation of Salmonella spp. in food, including seafood, adhere to a negative detection standard. The risk of infection is particularly high when seafood is consumed raw, which underscores the importance of timely detection of pathogenic microorganisms, such as Salmonella. Accordingly, this study aimed to develop a combined pre-treatment and detection method that includes pre-culture and DNA extraction in order to detect five species of Salmonella at concentrations below 10 CFU/mL in seafood. The effectiveness of the proposed method was assessed in terms of the composition of the enrichment (pre-culture) medium, minimum incubation time, and minimum cell concentration for pathogen detection. Furthermore, a practical DNA extraction method capable of effectively handling high salt conditions was tested and found to be successful. Through polymerase chain reaction, Salmonella spp. Were detected and positively identified in shellfish samples at cell concentrations below 10 CFU/g. Thus, the proposed method, combining sample pre-treatment and cell culture with DNA extraction, was shown to be an effective strategy for detecting low cellular concentrations of harmful bacteria. The proposed methodology is suitable as an economical and practical in situ pre-treatment for effective detection of Salmonella spp. in seafood.

Effects of Seed Specific Gravity by Germination and Seeding Emergence in Proso millet (Panicum miliaceum L.) (기장의 염수선 종자 정선법에 따른 발아 및 출현율 특성)

  • Jung, Ki-Youl;Choi, Young-Dae;Chun, Hyen-Chung;Lee, Sang-Hun;Shim, Sang-In;Jeon, Seung-Ho
    • Journal of the Korean Society of International Agriculture
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    • v.31 no.3
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    • pp.278-285
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    • 2019
  • The experiment was implemented to gain basic data for stable proso millet (Panicum miliaceum L.) production through germination viability and establishment uniformity based on seed refining under seed selection with specific gravity of proso millet seed. Seeds of three proso millet cultivars were carefully selected by adjusting NaCl into 9 levels (1.000, 1.005, 1.010, 1.015, 1.020, 1.025, 1.030, 1.035, 1.04 g·L-1), and their standard germination rate, seeding quality and field emergence rate were investigated. Proso millets showed heavier thousand seed weight as specific gravity high. In standard germination, all of the 3 cultivars of selected seeding showed at least 91% germination rate. With respect to the 1.040 g·L-1 selection by specific gravity, 'Hwanggeum' had 98.9%, 'Ibaekchal', 99.4%, and 'Hwangsilchal', 98.9%, recording the highest. Seeding quality was investigated and it was found that, compared with the non-selected seeds, selected seeds had longer plant heights and averaged 2.0, 2.2 and 1.9 cm, respectably. In terms of leaf length, only 'Hwanggeum' and 'Ibaekchal' showed distinctive differences. No significance was found in stem diameter and SPAD. The seeds selected using specific gravity all had longer or heavier average root length, root weight and stem weight. Compared with non-selected seeds, the specific gravity of 1.040 g·L-1 had longer root lengths of 2.9, 3.0 and 2.9 cm. 1.040 g·L-1 was also found to have the heaviest root weight, and stem weight. The field emergence viability investigation found significance solely in the mean emergence time (MET) by the cultivar. High significance was found in percent emergence (PE) and emergence rate index (ERI) in the specific gravity for the selection of seeds. All of the three cultivars showed higher PE and ERI as specific gravity selection high. The 1.040 g·L-1 specific gravity selection had the highest PE of 90.7, 85.3 and 92.8%. For productivity increase based on improved proso millet germination rate and enhanced establishment uniformity, it seems favorable to employ 1.040 g·L-1 specific gravity selection using salt solution.

Lithium Distribution in Thermal Groundwater: A Study on Li Geochemistry in South Korean Deep Groundwater Environment (온천수 내 리튬 분포: 국내 심부 지하수환경의 리튬 지화학 연구)

  • Hyunsoo Seo;Jeong-Hwan Lee;SunJu Park;Junseop Oh;Jaehoon Choi;Jong-Tae Lee;Seong-Taek Yun
    • Economic and Environmental Geology
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    • v.56 no.6
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    • pp.729-744
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    • 2023
  • The value of lithium has significantly increased due to the rising demand for electric cars and batteries. Lithium is primarily found in pegmatites, hydrothermally altered tuffaceous clays, and continental brines. Globally, groundwater-fed salt lakes and oil field brines are attracting attention as major sources of lithium in continental brines, accounting for about 70% of global lithium production. Recently, deep groundwater, especially geothermal water, is also studied for a potential source of lithium. Lithium concentrations in deep groundwater can increase through substantial water-rock reaction and mixing with brines. For the exploration of lithim in deep groundwater, it is important to understand its origin and behavior. Therefore, based on a nationwide preliminary study on the hydrogeochemical characteristics and evolution of thermal groundwater in South Korea, this study aims to investigate the distribution of lithium in the deep groundwater environment and understand the geochemical factors that affect its concentration. A total of 555 thermal groundwater samples were classified into five hydrochemical types showing distinct hydrogeochemical evolution. To investigate the enrichment mechanism, samples (n = 56) with lithium concentrations exceeding the 90th percentile (0.94 mg/L) were studied in detail. Lithium concentrations varied depending upon the type, with Na(Ca)-Cl type being the highest, followed by Ca(Na)-SO4 type and low-pH Ca(Na)-HCO3 type. In the Ca(Na)-Cl type, lithium enrichment is due to reverse cation exchange due to seawater intrusion. The enrichment of dissolved lithium in the Ca(Na)-SO4 type groundwater occurring in Cretaceous volcanic sedimentary basins is related to the occurrence of hydrothermally altered clay minerals and volcanic activities, while enriched lithium in the low-pH Ca(Na)-HCO3 type groundwater is due to enhanced weathering of basement rocks by ascending deep CO2. This reconnaissance geochemical study provides valuable insights into hydrogeochemical evolution and economic lithium exploration in deep geologic environments.

Characterization of the Behavior of Naturally Occurring Radioactive Elements in the Groundwater within the Chiaksan Gneiss Complex : Focusing on the Mineralogical Interpretation of Artificial Weathering Experiments (치악산 편마암 지질의 지하수 내 자연 방사성 원소의 거동 특성 연구: 인공풍화 실험을 통한 광물학적 해석)

  • Woo-Chun Lee;Sang-Woo Lee;Hyeong-Gyu Kim;Do-Hwan Jeong;Moon-Su Kim;Hyun-Koo Kim;Soon-Oh Kim
    • Korean Journal of Mineralogy and Petrology
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    • v.36 no.4
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    • pp.289-302
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    • 2023
  • The study area was Gangnim-myeon, Hoengseong-gun, Gangwon-do, composed of the Chiaksan gneiss complex, and it was revealed that the concentrations of uranium (U) and thorium (Th) within the groundwater of the study area exceeded their water quality standards. Hence, artificial weathering experiments were conducted to elucidate mineralogically the mechanisms of their leaching using drilling cores obtained from the corresponding groundwater aquifers. First of all, the mineralogical compositions of core samples were observed, and the results indicated that the content of clinochlore, a member of the chlorite group of minerals that can form through low- and intermediate-temperature metamorphisms, was relatively higher. In addition, the Th concentration was measured ten times higher than that of U. The results of artificial weathering experiments suggested that the Th concentrations gradually increased through the dissolution of radioactive-element-bearing minerals up to the first day, and then they tended to decrease. It could be attributed to the fact that Th was leached with the dissolution of thorite, which might be a secondary mineral, and then dissolved Th was re-precipitated as the various forms of salt, such as sulfate. Even though the U content was lower than that of Th in the core samples, the U concentration was one hundred times higher than that of Th after the weathering experiments. It is likely caused by the gradual dissolution and desorption of U included in intensively weathered thorite or adsorbed as a form of UO22+ on the mineral surface. In addition, the leaching tendency of U and Th was positively correlated with the bicarbonate concentration. However, the concentrations between U and Th in groundwater exhibited a relatively lower correlation, which might result from the fact that they occurred from different sources, as aforementioned. Among various kinetic models, the parabolic diffusion and pseudo-second-order kinetic models were confirmed to best fit the dissolution kinetics of both elements. The period that would be taken for the U concentration to exceed its drinking-water standard was inferred using the regressed parameters of the best-fitted models, and the duration of 29.4 years was predicted in the neutral-pH aquifers with relatively higher concentrations of HCO3, indicating that U could be relatively quickly leached out into groundwater.

The acute toxicity and efficacy evaluation against Aeromonas salmonicida of aquatic drugs oxolinic acid, neomycin-oxytetracycline, and florfenicol in guppy (Poecilia reticulata) (구피(Poecilia reticulata)에서 수산용의약품 oxolinic acid, neomycin-oxytetracycline, florfenicol의 급성독성 및 Aeromonas salmonicida에 대한 약효 평가)

  • Jun Sung Bae;Chae Won Lee;Chan Yeong Yang;Eun Ha Jeong;Areum Kim;Young-Sik Chae;Jung-Jin Park;Kwan Ha Park
    • Journal of fish pathology
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    • v.36 no.2
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    • pp.293-302
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    • 2023
  • This study evaluated the acute toxicity induced by oxolinic acid (OA), neomycin-oxytetracycline combination (NEO-OTC) and florfenicol (FF) administered to guppy (Poecilia reticulata) by comparing standard formulations with commercial preparations (COOaqua curinpotion, COOaqua terafinpotion and COOaqua profenpotion, respectively) for ornamental fish at concentrations of 2-4%. NEO-OTC exhibited the highest acute toxicity in guppy, with no difference observed between the pstandard formulation and commercial preparation (LC50 = 126.08 mg/L and 112.44 mg/L, respectively). OA acute toxicity was assessed in the form of sodium salt, with an LC50 of 504.61 mg/L for the standard formulation and a slightly increased toxicity of 316.11 mg/L for the commercial preparation. In contrast, no mortality was observed during the 96-hour exposure period with the standard formulation in the form of oxolinic acid. The acute toxicity of FF was measured to be above 1,000 mg/L for the standard formulation; however, the commercial preparation significantly increased to 158.53 mg/L. These results indicate that toxicity can significantly increase in commercial formulations, especially those with low levels of active ingredients. This is presumed to be attributed to the organic solvents or solubilizing agents present in the commercial preparations, which may enhance toxicity. Additionally, guppy infected with Aeromonas salmonicida were effectively protected against mortality by administering OA, NEO-OTC and FF at concentrations of 50 mg/L, 100 mg/L and 15 mg/L, respectively, for 2 hours and at half the dose for 24 hours. This result indicates that liquid formulations containing low concentrations of antibiotics may partially increase toxicity, but there is no problem in effectively treating diseases in ornamental fish.

Characterization of typical Aeromonas salmonicida isolated from Sea-Chum Salmon (Oncorhynchus keta) (해수에 순치된 첨연어(Oncorhynchus keta)에서 분리된 정형 에로모나스 살모니시다(Aeromonas salmonicida)에 대한 특성 분석)

  • Jongwon Lim;Sungjae Ko;Youngjun Park;Do-il Ahn;Suhee Hong
    • Journal of fish pathology
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    • v.36 no.2
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    • pp.263-275
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    • 2023
  • Chum salmon (Oncorhynchus keta) is a species which returns to Korea for spawning and was produced as seed production at the Fisheries Resources Agency located in Uljin-gun, Gyeongsangbuk-do to preserve the species. However, farmed chum salmon showed symptoms of bacterial infection. Therefore, in this study, bacteria were isolated to identify the causative agent from chum salmon in October 2021. The isolated bacteria were identified based on the sequences of 16S rDNA, rpoD (RNA polymerase sigma factor σ70), and vapA (A-layer) genes. Also, salinity-growth curve, biochemical characterization, antibiotic susceptibility test, and pathogenicity analysis were performed in four strains. As a result, four isolated strains were identified as Aeromonas salmonicida subsp. salmonicida. Additionally, the bacterial strains showed a decrease in growth as the salt concentration increased in the medium. All of the isolated strains exhibited γ-hemolysis, and the same biochemical properties. In the antimicrobial susceptibility test, all strains showed an inhibition zone of 40 to 44 mm for oxolinic acid, flumequine, and florfenicol. Pathogenic factors were assessed by RT-PCR at the mRNA level, and found that the four strains expresses the outer membrane ring of T3SS (ascV), inner membrane ring of T3SS (ascC), vapA, enterotoxin (act), and lipase (lip) genes which are well known to significantly contribute to the pathogenicity of A. salmonicida. The results of this study can be used as basic data to prevent A. salmonicida subsp. salmonicida occurring in sea-chum salmon in the future.

Effect of Fractions From Stachys sieboldii Miq. Root on Antioxidant, Anti-inflammation and Smad Signaling (초석잠 뿌리 분획물의 항산화 및 항염증 효과와 smad 신호 전달에 미치는 효과)

  • Jung Woo Lee;Myungwon Choi;Sun Young Lim
    • Journal of Life Science
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    • v.34 no.4
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    • pp.245-253
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    • 2024
  • We investigated to analyze total flavonoid content and fatty acid composition of Stachys sieboldii Miq root. In order to determine antioxidant and anti-inflammatory effects of fractions from S. sieboldii Miq. root, we conducted 1.1-Diphenyl-2-picryhydrazyl (DPPH) and 2,2'-Azino-bis (3-ethylbenothiazoline-6-sulfonic acid) diammonium salt radical cation (ABTS) assays for antioxidant and measured nitric oxide (NO) production induced by lipopolysaccharide (LPS) in RAW 264.7 cells. In addition, we examined an inhibitory effect of fractions from S. sieboldii Miq. root on smad signaling induced by transforming growth factor (TGF) β. Among the fractions, n-butanol (n-BuOH) fraction showed the highest flavonoid content (16.67 mg/g), followed by n-Hexane, water and 85% aqueous methanol (85% aq. MeOH) fractions. The fatty acid composition of S. sieboldii Miq. root was in the following order : n-6 fatty acids (54.3%) > n-3 fatty acids (21.2%) > saturated fatty acids (19.7%) > n-9 fatty acids (3.6%). As a result of the antioxidant efficacy, the DPPH and ABTS assays showed that n-BuOH fraction had higher scavenging activity compared to other fractions. Inhibitory effect on NO production showed that all fractions decreased LPS-induced NO production, indicating an anti-inflammatory activity of S. sieboldii Miq. root. 85% aq. MeOH and water fractions showed a higher efficacy in inhibiting transforming growth factor (TGF) β induced smad signaling. From the results, it suggests that food processing products using S. sieboldii Miq. root will be developed as a functional food for promoting health.

Study on Forage Cropping System Adapted to Soil Characteristics in Reclaimed Tidal Land (간척지 토양특성에 알맞은 사료작물 작부체계 연구)

  • Yang, Chang-Hyu;Lee, Jang-Hee;Kim, Sun;Jeong, Jae-Hyeok;Baek, Nam-Hyun;Choi, Weon-Young;Lee, Sang-Bok;Kim, Young-Doo;Kim, Si-Ju;Lee, Gyeong-Bo
    • Korean Journal of Soil Science and Fertilizer
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    • v.45 no.3
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    • pp.385-392
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    • 2012
  • This study was conducted to find out the optimum cropping system for the stable production of forage crops in the newly reclaimed land located at Gwanghwal and Gyehwa region of Saemangum reclaimed tide land from October, 2009 to October, 2011. Whole crop barley (WCB), Rye, Italyan-ryegrass (IRG) as winter crops and Corn, Sorghum${\times}$sudangrass hybrid (SSH) as summer crops were cultivated. Soil chemical properties, nutrient uptake, feed value, growth and yield were examinated. The testing soil was showed saline alkali soil where the contents of organic matter, available phosphate and exchangeable calcium were very low, while exchangeable sodium and magnesium were higher. Changes of soil salinity during the growing season of forage crops were less than 0.2%, and the growth of forage crops was not affected by salt injury. Standing rates of winter crops were higher in the order of Rye, WCB, and IRG, while the dry matter yield of winter crops was higher in the order of IRG, Rye and WCB. The highest crude protein (CP) content was recorded in IRG, and total digestive nutrients (TDN) contents were increased in the order of WCB, IRG, and Rye. The TDN content was higher in corn, whereas other feed value was higher in SSH. The content of mineral nutrients on stem, leaf and grain in IRG, Corn were high. After experiment pH was lowed, contents of exchangeable magnesium, sodium and organic matter were decreased while contents of total nitrogen, available phosphate and exchangeable potassium, calcium were increased. Winer crops and summer crops after continually cultivating in cropping system, fresh matter yield increased, compared to WCB-Corn (74,740 kg $ha^{-1}$), IRG-SSH 10%, IRG-Corn 7%, Rye-SSH 6%, Rye-Corn and WCB-SSH 3%. Dry matter yield increased, compared to WCB-Corn (20,280 kg $ha^{-1}$), IRG-SSH 7%, Rye-SSH 6%, IRG-Corn/Rye-Corn/WCB-SSH 3%. The TDN yield increased, compared to WCB-Corn (13,830 kg $ha^{-1}$), IRG-SSH 2%, WCB-SSH and IRG-Corn 1%. Therefore, we suggest that the crop combination of IRG-SSH and WCB-SSH would be preferred for silage stable production.

Studies on the analysis of phytin by the Chelatometric method (Chelate 법(法)에 의(依)한 Phytin 분석(分析)에 관(關)한 연구(硏究))

  • Shin, Jai-Doo
    • Applied Biological Chemistry
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    • v.10
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    • pp.1-13
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    • 1968
  • Phytin is a salt(mainly calcium and magnesium) of phytic acid and its purity and molecular formula can be determined by assaying the contents of phosporus, calcium and magnesium in phytin. In order to devise a new method for the quantitative analysis of the three elements in phytin, the chelatometric method was developed as follows: 1) As the pretreatment for phytin analysis, it was ashfied st $550{\sim}600^{\circ}C$ in the presence of concentrated nitric acid. This dry process is more accurate than the wet process. 2) Phosphorus, calcium and megnesium were analyzed by the conventional and the new method described here, for the phytin sample decomposed by the dry process. The ashfied phytin solution in hydrochloric acid was partitioned into cation and anion fractions by means of a ration exchange resin. A portion of the ration fraction was adjusted to pH 7.0, followed by readjustment to pH 10 and titrated with standard EDTA solution using the BT [Eriochrome black T] indicator to obtain the combined value of calcium and magnesium. Another portion of the ration fraction was made to pH 7.0, and a small volume of standard EDTA solution was added to it. pH was adjusted to $12{\sim}13$ with 8 N KOH and it was titrate by a standard EDTA solution in the presence of N-N[2-Hydroxy-1-(2-hydroxy-4-sulfo-1-naphytate)-3-naphthoic acid] diluted powder indicator in order to obtain the calcium content. Magnesium content was calculated from the difference between the two values. From the anion fraction the magnesium ammonium phosphate precipitate was obtained. The precipitate was dissolved in hydrochloric acid, and a standard EDTA solution was added to it. The solution was adjusted to pH 7.0 and then readjusted to pH 10.0 by a buffer solution and titrated with a standard magnesium sulfate solution in the presence of BT indicator to obtain the phosphorus content. The analytical data for phosphorus, calcium and magnesium were 98.9%, 97.1% and 99.1% respectively, in reference to the theoretical values for the formula $C_6H_6O_{24}P_6Mg_4CaNa_2{\cdot}5H_2O$. Statical analysis indicated a good coincidence of the theoretical and experimental values. On the other hand, the observed values for the three elements by the conventional method were 92.4%, 86.8% and 93.8%, respectively, revealing a remarkable difference from the theoretical. 3) When sodium phytate was admixed with starch and subjected to the analysis of phosphorus, calcium and magnesium by the chelatometric method, their recovery was almost 100% 4) In order to confirm the accuracy of this method, phytic acid was reacted with calcium chloride and magnesium chloride in the molar ratio of phytic: calcium chloride: magnesium chloride=1 : 5 : 20 to obtain sodium phytate containing one calcium atom and four magnesium atoms per molecule of sodium phytate. The analytical data for phosporus, calcium and magnesium were coincident with those as determine d by the aforementioned method. The new method employing the dry process, ion exchange resin and chelatometric assay of phosphorus, calcium and magnesium is considered accurate and rapid for the determination of phytin.

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Soil amendment for turfgrass vegetation of the Incheon International Airport runway side on the Yeongjong reclaimed land (인천국제공항 착륙대 잔디 식재 지반 조성을 위한 영종도 매립 토양 개량)

  • Yoo, Sun-Ho;Jeong, Yeong-Sang;Joo, Young-Kyu;Choi, Byung-Kwon;Wu, Heun-Young;Lee, Tae-Young
    • Korean Journal of Soil Science and Fertilizer
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    • v.35 no.2
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    • pp.93-104
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    • 2002
  • A field survey and experiment was conducted from 1996 to 1998 to develop rational technology for turfgrass vegetation of runway side of Incheon International Airport on the reclaimed tidal land in Young-Jong Island. Backfill of the experimental site was finished on August 1995. The experimental site was 8 ha located in the middle of the construction place for the main parking lot in front of the terminal building construction. The experimental field was drained by main open ditch, and divided three main plots, no subsurface tile drain, subsurface tile drain spacing with 22.5m, and with 45 m, respectively. The 17 sub plots were designed to test the effect of soil covering with red earth loam by 5 cm and 20 cm depth, application of chemical compound fertilizers and livestock manures, dressing of artifical soils and hydrophylic soil conditioners. The tested turfgrasses were three transplanting indigenous turfgrasses, Zoysia koreana, Zoysia sinica and Zoysia japonica, and two hydroseeding mixed exotic turgrasses, cool type I(tall fescue 30%, kentucky blue grass 40%, perenial ryegrass 30%), and cool type II(tall fescue 40%, perenial ryegrass 20%, fine fescue 20%, alkaligrass 20%). The soil backfilled with dredged seasand was sand textured with high salt concentration and low fertility. The soil showed high pH, low organic matter and low available phophate contents. The percolation rate was fast with high hydraulic conductivity. Desalinization was fast after installation of the main open drainage system. No subsurface tile drainage effect was found showing little difference in turfgrass growth. The covering and visual growth of turfgrasses were the best in the 20-cm soil covering with compound fertilizer treatment. The covering and visual growth of turfgrasses were satisfactory in the 5 cm soil covering with compound fertilizer treatment and with livestock manure treatments. The hydrophillic soil conditioner treatments were effective but expensive at present. The coverage and visual quality of turfgrasses were good for Zoysia koreana and Zoysia japonica. The coverages of turfgrasses by the hydroseeding with the mixed exotic turfgrasses were less than transplanting of native turfgrasses. In conclusion, for the runway side vegetation purposes, the subsurface tile drainage might not necessary as main open ditch drainage be sufficient due to fast percolation rate of the backfilled dredged seasand. The 5 cm soil covering with red earth might be sufficient for the runway side, but the 20 cm soil covering might be necessary for the runway side where high density of turfgrass coverage was necessary to protect from the airplance air blow.