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

• 원예과학기술지
• /
• 제17권3호
• /
• pp.337-340
• /
• 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
• /
• 제19권4호
• /
• pp.223-231
• /
• 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
• /
• 제24권2호
• /
• pp.147-153
• /
• 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.

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

• 한국가축번식학회지
• /
• 제21권3호
• /
• pp.211-217
• /
• 1997

This experiment was carried out to determine the possibility of using FDA on the selection of viable bovine immature oocytes. The results obtained in these experiments were summarized as follows; 1. The rates of nuclear maturation (metaphase II) of bovine oocytes according to FDA concentration were 92.5% (37/40) in the control and those rates in the dilution on FDA to 1:400,000, 1:800,000 and 1:1,600,000 were 74.4% (67/90), 80.3% (38/46) and 71.1% (32/45), respectively. 2. The fertilization rate ( 2-cell) in the control was 72.2% (39/54) and those rates in the dilution of FDA to 1:400,000, 1:800,000 and 1:1,600,000 were 28.8% (23/80), 66.7% (32/48) and 62.2% (28/45), respectively. In these results, a, pp.opriate concentration of FDA to bovine immature oocytes was 1:800,000. 3. Effect of FDA treatment to the blastocyst development of bovine oocytes was indicated that control was 22.2% (18/81) and FDA treatment groups which were classified to strong, partial and weak were 21.4% (36/168), 14.5% (9/62) and 0% (0/15), respectively. This result suggested that in vitro development to the blastocyst was severely reduced except strong group according to the FDA fluorescent level (P<0.05) and that using FDA is possible to select of bovine oocytes.

• 한국수정란이식학회지
• /
• 제10권3호
• /
• pp.193-202
• /
• 1995

In order to improve the cryopreservatory techniques of livestock embryos, the quick freezing method which is directly plunged in liquid nitrogen via prefreezing procedure without freezing machine was carried out for mouse embryos treated with permeable and nonpermeable cryoprotectants. The viability of frozen-thawed embryos were evaluated by FDA vital dye test. The results obtained was summaried as follows: 1. A total of 720 embryos were recovered from frozen embryos for viability test. Evalution of the fluorescein diacetate(FDA) vital dye test with mice embryos were resulted of 2.3 total mean score - evaluted in orderly higher mean grade of P3 453 (63%), P2 133(18%), P1 51(7%) and P0 83(12%). 2. An all-round evalution of these combination, the highest viability was showed in 3M ethylene glycol + 0. 25M trehalose treated with the copper prefreezing. 3. Effects of permeable and nonpermeable cryoprotectants combination were evaluated by means FDA score. 3M ethylene glycol + 0.25M trehalose showed the highest survival rates of 2.8 mean FDA score. 4. Effects of permeable cryoprotectants were evaluated by mean FDA score but the results were not significantly different each other. 5. In evalution of the nonpermeable cryoprotectants, 0. 25M trehalose obtalned higher mean FDA score than of 0.25M sucrose and it was significantly different(P<0.05). 6. There was no significantly difference between copper and stainless-steel in prefreezing procedures.

• 한국토양비료학회지
• /
• 제42권4호
• /
• pp.307-316
• /
• 2009

토양 개량을 위해 시용하는 유기물은 종류에 따라서 토양미생물 활성과 다양성에 미치는 영향을 다르며 그 효과는 계절적인 영향을 크게 받는다. 도시 가로수 폐기물 퇴비, 가금류 분뇨, 귀리와 레드 클로버의 피복작물이 토양 효소활성과 토양 미생물의 생리적 군락(CLPP) 특성에 미치는 영향을 미국 미주리 주의 사양토에서 조사하였다. 이들 토양 조사 항목들의 계절적 변화 패턴을 조사하기 위해서 2년간 봄부터 가을까지 토양을 매년 5회 채취하였다. 나무 폐기물 퇴비는 시용 3달 후부터 탈수소효소의 활성을 증가시키기 시작하였다. fluorescein diacetate (FDA) 수화도는 첫 해의 9월부터 증가하기 시작했으나 그 이후 변화가 심하였다. 탈수수효소의 활성은 FDA 소화도에 비하여 토양 유기물의 량이나 특성에 더 직접적으로 반응하였다. 반면에 FDA 수화도나 CLPP는 일반적으로 유기물의 구성 성분에 반응하였고, 효소활성과 CLPP 모두 계절에 따른 변화가 심하였다. 계절에 따른 변화는 유기물과 토양 수분함량의 차이에 기인한 것으로 보였다. 도시 가로수 폐기물 퇴비는 일반적인 토양 미생물 활성을 증가시키는데 효과적이었고 녹비는 토양 미생물 군락의 다양성을 변화시키는데 효과적이었다. 그리고 토양 미생물 활성과 다양성은 계절적 변화가 심하고 그 정도는 시용하는 유기물의 종류에 따라서 차이가 있으므로 토양의 미생물 특성을 조사할 때에는 작물의 재배기간 동안 여러 번 실시할 필요가 있다.

• 한국가축번식학회지
• /
• 제18권1호
• /
• pp.55-62
• /
• 1994

This study was carried out to test the effects of fluorescein diacetate(FDA 0${mu}ell$/ml, 0.5${mu}ell$/ml, 5${mu}ell$/ml, 10${mu}ell$/ml or 0${mu}ell$/ml, 5${mu}ell$/ml in PBS) treated before culture on the survival of vitrified mouse morulae in vitrification solution(20% glycerol＋glycerol＋10% ethylene glycol＋30% ficoll＋10% sucrose). The results were summarized as follows; 1. The survival rate of FDA-tested fresh mouse morulae after 24 hours culture was over 96%((P4.8) in the control or treatment groups with various levels of FDA. Because the rate of mouse morulae which developed to hatched blastocysts was higher with the various levels of FDA treatment(67%) than control(50%), it was considered that toxicity of FDA did not affect the survival of mouse morulae. 2. When mouse morulae were FDA-tested in FDA 0(control), 0.5, 5, and 10${mu}ell$/ml treatment after vitrification, the development rate to expanded blastocyst were 66, 82, 64 and 76%, and FDA scores were P4.2(84%), P4.7(94%), P4.2(84%) and P3.9(78%), respectively. There were no significant differences between control and FDA treatments, but there were significant difference between 0.5${mu}ell$/ml)94%) and 10${mu}ell$/ml(78%) treatment(P<0.01). 3. The survival rates of cultured mouse morulae according to FDA-scores(P0=non-fluorescence; P1~P5=according to their fluorescence) after vitrification were P5;92%, P4;67%, P3;42% and P2.P0;0%, respectively. 4. Implantation rates of morulae stage embryos cultured into early blastocysts and implanted into uterine hornes vitrification were 14 and 11% embryos treated control(0${mu}ell$/ml) and FDA 5${mu}ell$/ml and the normal fetus development was 2% embryos for both treatments. Results of this percent study indicated that toxicity of FDA does not affect not only the survival of fresh and vitrified mouse morulae but also the development rate and implantation of fetus after transfer as well. The development rates of mouse morulae with the FDA score of P5, P4 and P3 were 92, 67 and 42%, respectively, it was considered that FDA-test was fit for the judge of survival.

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