• Ocean and Polar Research
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• 제31권4호
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• pp.349-357
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• 2009

Ballast water is widely recognized as a serious environmental problem due to the risk of introducing non-indigenous aquatic species. In this study we aimed to investigate measures which can minimize the transfer of aquatic organisms from ballast water. Securing more reliable technologies to determine the viability of aquatic organisms is an important initiative in ballast water management systems. To evaluate the viability of marine phytoplankton, we designed the staining methods of fluorescein diacetate (FDA) and Calcein-AM assay on each target species belonging to different groups, such as bacillariphyceae, dinophyceae, raphidophyceae, chrysophyceae, haptophyceae and chlorophyceae. The FDA method, which is based on measurements of cell esterase activity using a fluorimetric stain, was the best dye for determining live cells of almost all phytoplankton species, except several diatoms tested in this study. On the other hand, although fluorescence of Calcein-AM was very clear for a comparatively longer time, green fluorescence per cell volume was lacking in most of the tested species. According to the Flow CAM method, which is a continuous imaging technique designed to characterize particles, green fluorescence values of stained cells by FDA were significantly higher than those of Calcein-AM treatments and control, implying that the Flow CAM using FDA assay could be adapted as an important tool for distinguishing living cells from dead cells. Our results suggest that the FDA and Calcein-AM methods can be adapted for use on phytoplankton, though species-specific characters are greatly different from one organism to another.

• 미생물학회지
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• 제24권2호
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• pp.147-153
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• 1986

나균(M. leprae)을 비롯한 항산균(mycobacterium)의 시험관내 생리력 또는 생명력평가법(in vitro assessment of physiological potential or viability)으로 형광분광측정법(fluorospectrophotometry)를 고안하였다. 균액을 비형광성의 fluorescein diacetate(FDA)로 처리, 균체의 생대사능에 의해 FDA로부터 유리된 체내 fluorescein 량을 Aminco-Bowman 형광분광기로 측정함으로 균체의 생리능을 fluorounit로 표기해 보았다. Fluorounit로 표기된 균체의 생명력을 균배양의 광학밀도(optical density, colony forming unit, 체내 ATP 량 intracellular ATP content)등으로 표기되는 항산균의 생명력고 비교 검토함으로 형광분광법에 의한 시험관내 항산균의 생명력 평가법의 적합성을 살펴보았다. 형광분광측정에 의한 생명도의 평가는 객관성을 띤 기기정량법으로 조작이 간단하고 신속하게 결과를 얻을 수 있어 시험관내 배양의 속도가 완만한 균이나(slow growing mycobacteria, I.e.M. lerpae, Listeriae sp)등의 생명력의 상대적 평가에 활용될 수 있다고 사료된다.

• Weed & Turfgrass Science
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• 제3권4호
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• pp.354-361
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• 2014

This study was conducted to investigate whether several composted liquid manures (CLMs) are useful for biological control of large patch on zoysiagrass and investigate the chemical and biological factors to suppress large patch in soil treated with CLMs. The CLMs were produced at 4 different facilities for livestock excretion treatments located in Korea. Field experiments were carried out at 5 golf courses located near each facility. CLM and Chemical fertilizer (CF: water soluble fertilizer, 20-20-20) were applied four and three times with N at $12g\;m^{-2}$ per year, respectively. There was significant increase of concentration of K, Na, and Cu of soil treated with CLM compared to CF treatment. Among experimental plots, CN and GG2 plot sites were shown significant higher effect of biological control 80% and 50% respectively against large patch disease. The number of bacteria, Actinomycetes, and fungi in soil at these sites significantly increased and fluorescein diacetate hydrolytic activity was enhanced, while the soil was treated with CLM. The results of this study demonstrated that CLM application has effect on soil to suppress large patch and reduce the use of fungicide in environment-friendly turf management.

• 원예과학기술지
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• 제17권3호
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• pp.337-340
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• 1999

장기 저장한 사과 화분의 활력을 측정하기 위하여 기본배지와 fluorescein diacetate(FDA)를 첨가한 배지를 slide glass 위에 얇게 도포한 후 기내발아율을 조사한 결과, FDA를 첨가한 배지에서는 화분 발아가 저하되었다. 따라서 FDA 용액을 이용한 형광염색반응법으로 사과 화분의 활력을 측정할 수 있는지 구명하기 위하여 10%의 sucrose 용액을 slide glass 위에 한방울 떨어뜨린 후 0.002%의 FDA를 첨가한 곳에 화분을 치상한 결과, 형광반응이 일어나는 치상 후 30분 경에 활력이 있는 화분은 자외선하에서 명확히 형광을 나타냈다. 기내발아율과 형광반응률을 측정한 결과, 39개월 장기 저장한 사과의 화분도 활력이 감소하지 않는 것으로 나타났다. 같은 방법으로 조사한 품종별 화분의 활력은 '후지' 와 '천추'가 가장 높게 나타났으며 '쓰가루'는 중간 정도, '조나골드'는 낮게 나타났다. 형광염색반응법에 의한 형광반응률은 대체로 기본배지에서의 발아율과 비슷하게 나타났으나 3개월간 저장한 '천추'의 화분은 형광반응률이 발아율보다 다소 높게 나타났다. 형광반응률과 발아율은 높은 정의 상관을 보여 형광염색 반응법에 의해 장기 저장 사과 화분의 활력을 편리하고 정확하게 측정할 수 있었다.

• 한국해양학회지:바다
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• 제19권4호
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• pp.223-231
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• 2014

선박평형수를 통한 외래종 확산을 방지하기 위해 국제해사기구(IMO)에서는 2004년 선박평형수관리협약을 채택하였다. 이 협약에 따르면 앞으로 대부분의 선박들은 선박평형수 처리시스템을 통해서 해양 생물을 사멸 또는 제거시킨 후 배출해야 하며, 이는 플랑크톤의 생사판별방법을 통하여 이루어진다. 본 연구에서는 국제적으로 선박평형수 처리후 생사판별법으로 널리 사용되고 있는 fluorescein diacetate assay (FDA) 염색방법의 제한성과 이의 대안으로 Neutral red (NR) 염색방법사용 가능성을 살펴보고자 하였다. FDA 염색법은 대부분의 플랑크톤 염색에 되어서 현재 가장 널리 사용되는 방법임에도 불구하고 본 연구에서는 Ditylum brightwellii 을 제외한 모든 식물플랑크톤 대해서 낮은 염색 효율(전체 평균 염색 효율 <50%)을 보였으며, 식물플랑크톤이 갖는 고유한 형광(적색)에 간섭을 많이 받는 것으로 나타났다. 또한 FDA는 형광파장에 노출된 후 빠르게 색이 바래지는 경향도 관찰되었다. 반면에 NR은 조사된 모든 동 식물플랑크톤 개체수에 대해서 90% 이상의 높은 염색 효율을 보였다. 두 염색법 모두 포르말린을 이용해서 사멸시킨 동 식물플랑크톤에 대해서는 염색이 되지 않았다. 본 연구결과를 통해서 NR 염색법을 이용한 동 식물플랑크톤 생사판별은 매우 효율적이라고 판단된다.

• 미생물학회지
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• 제32권2호
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• pp.163-171
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• 1994

경유를 sandy loam 토양에서 농도별로 투여하여 토양미생물군집에 미치는 영향을 조사하였으며 또한 경우가 첨가된 토양에서 경유의 신속한 제거를 위해 실시한 bioremediation에 의한 영향을 측정하였다. 경유는 토양에서 16주 후 약 50% 내외의 잔류량을 나타내며 bioremediation 처리시 제거율은 60~95% 정도가 증가하여 16주 후 약 8~20 범위의 잔류량을 나타내었다. 토양에 경유가 첨가되었을 때에는 세균직접계수, 진균류의 균사 길이, 호기성 종속영양세균과 탄화수소 분해세균의 수가 최고 10 내지 100배 정도 증가하였다. Bioremediation 처리시에는 그 증가가 더욱 두드러져 각종 미생물 개체수 측정치가 최고 100배 내지 1000배까지 증가하였다. 경유가 fluorescein diacetate 가수분해 활성에 미치는 영향은 bioremediation 처리를 하지 않은 토양에서는 뚜렷한 증감의 경향이 없었으나 bioremediation 처리를 한 토양에서는 10배 내외의 활성의 증가를 보였으며 이러한 양상은 soil dehydrogenase 활성에서도 유사하게 나타났다.

• Clinical and Experimental Reproductive Medicine
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• 제19권1호
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• pp.1-8
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• 1992

A fluorescence microscopy technique using flurescein diacetateCFDA) as a substract has been tested for the evaluation of the viability of early mouse embryos. Embryos were incubated in T6 containing FDA concentrations of 2.5 to $50{\mu}g/ml$ for 1 to 5min. Embryos were then examined by reflected light fluorescence using a KP 490 and 520 barrier filter in a Nicon Diaphot microscopy. The results were as follow. 1. The rate of fluorescein accumulation increased on the concentration on FDA from $2.5{\times}10^{-6}M$ to $20{\times}10^{-6}M$ 2. The rate at which intracellular fluorescein was lost from embryos was depended on the temperature at which are stored. 3. Embryos with 3 min exposure to FDA have the most intensity of fluorescence. 4. Exposure of 2 cell embryos to FDA ($2.5-5{\mu}g/ml$) for 1 min did not alter their ability to delope normally in vitro.

• Natural Product Sciences
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• 제7권3호
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• pp.72-75
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• 2001

The antioxidant activity of roasted defatted perilla (Perilla frutescens) seed was determined by measuring its radical scavenging effect on 1,1-diphenyl-2- picrylhydrazyl (DPPH) radicals, inhibitory activity on total reactive oxygen species generation in kidney homogenates using 2',7'-dichlorodihydro-fluorescein diacetate, and scavenging effect on authentic peroxynitrites. The methanolic extract of roasted defatted perilla seed showed strong scavenging activity in both DPPH and peroxynitrite radicals, and thus fractionated with several solvents. The antioxidant activity potential of the individual fraction was in the order of ethyl acetate>n-butanol>dichloromethane>water>n-hexane fraction. The ethyl acetate soluble fraction exhibiting strong antioxidant activity was further purified by repeated silica gel and Sephadex LH-20 column chromatography. Luteolin was isolated as one of the active principles from the ethyl acetate fraction, together with the inactive chrysoeriol and apigenin.

• 한국지하수토양환경학회:학술대회논문집
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• 한국지하수토양환경학회 2000년도 추계학술대회
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• pp.196-201
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• 2000

The influence of adsorption on cadmium toxicity to soil microorganisms in smectite-rich soils and sediments was quantified as a function of solution and sorbent characteristics. Adsorption and surface complexation experiments were conducted to infer Cd sorption mechanisms to a reference smectite and three fractions of a Veritsol soil, and to elucidate the effects of the surface complexation on Cd bioavailability and toxicity in soils and sediments. Cadmium adsorption isotherms conformed to the Langmuir adsorption model, with adsorptive capacities of the different samples dependent on their characteristics. Equilibrium geochemical modeling (MINTEQA2) was used to predict the speciation of Cd in the soil suspensions using Langmuir and Triple Layer surface complexation models. The influence of adsorption and surface complexation on cadmium toxicity to soil microorganisms was assessed indirectly through the relative change in microbial hydrolysis of fluorescein diacetate (FDA) as a function of total Cd concentration and sorbent characteristics. Adsorption decreased the toxicity of Cd to soil microorganisms. Inner-sphere complexation is more effective than outer-sphere complexation in reducing the bioavailability and toxicity of heavy metals in soils and sediments.

• Environmental Engineering Research
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• 제22권2호
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• pp.224-229
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• 2017

Zinc oxide nanoparticles (ZnO NPs) have been used as additives in a variety of consumer products. While these particles may enter the environment, only a limited number of studies have investigated the effects of ZnO NPs on soil exoenzymes. Here, we investigate the long-term effects of ZnO NPs at concentrations of 50 and 500 mg/kg on the activities of six soil exoenzymes in planted soils: Dehydrogenase, fluorescein diacetate (FDA) hydrolase, urease, acid phosphatase, arylsulfatase, and ${\beta}-glucosidase$. Significant effects were observed at one or more time points for all enzymes except for FDA hydrolase. These effects included both decreases and increases in enzyme activity. Our results suggest that ZnO NP treatments of 50 and 500 mg/kg can adversely affect soil enzymes, particularly acid phosphatase and urease, and thus, these data may have implications for phosphorous and nitrogen cycles in the soil.