• Journal of Environmental Science International
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• v.15 no.10
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• pp.961-966
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• 2006

An algal assay procedure using an indigenous phytoplankton assemblage was tested to estimate the propagation of red tide phytoplankton species and determine the optimal time interval at which to measure growth yield in eutrophic marine waters where red tides frequently occur. Various red tide phytoplankton species were propagated on a large scale by adding nitrogen or phosphorous. This procedure was useful for estimating the limiting nutrient, elucidating the mechanisms underlying red tides, and determining the levels of increases in organic matter in eutrophic coastal waters. The algal assay using indigenous C. polykrikoides showed that this species did not always propagate, apparently because of very low concentrations of trigger elements that are necessary for its growth, rather than as a result of other environmental characteristics, e.g., water temperature or stress from sampling. In the winter, when water temperatures are lower than in spring, summer, or autumn, maximum propagation and the limiting nutrient could be estimated by measuring phytoplankton biomass at 2 - 3-day intervals. However, in the other seasons, when water temperatures are higher, phytoplankton biomass should be measured at 2-day intervals. In particular, daily monitoring will be required to determine precise growth yields in warm seasons.

• Ocean and Polar Research
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• v.31 no.4
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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.

• The Sea:JOURNAL OF THE KOREAN SOCIETY OF OCEANOGRAPHY
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• v.19 no.4
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• pp.223-231
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• 2014

In order to prevent the spread of non-indigenous aquatic species through the ballast water in commercial ships, International Maritime Organization (IMO) adopted in 2004 the International Convention for Control and Management of Ship's Ballast Water and Sediments. The Convention mandates treatment of ballast water for most transoceanic voyages and its confirmation of treatment is made with plankton live/dead assay. Fluorescein diacetate assay (FDA), which produces bright green light for live phytoplankton, has been a de facto standard method to determine the survival of marine plankton, but its staining efficacy has been in dispute. In the present study, we examined the limitation of FDA, and compared its efficacy with Neutral red (NR) staining, another promising assay and widely used especially for zooplankton mortality. For all phytoplankton species studied in the present study, except Ditylum brightwellii, the staining efficiency was <50% with FDA. The green FDA fluorescence interfered with phytoplankton autofluorescence in most samples. In contrast, NR assay stained over 90% of both phytoplankton and zooplankton species tested in this study. FDA assay also showed that green FDA fluorescence rapidly faded when phytoplankton cells were exposed to microscope light. Both FDA and NR assay were negative on formalin-killed individuals of both phytoplankton and zooplankton species. Our results suggest that NR assay is more effective for determining the survival of marine plankton and can be applied to test the efficacy of ballast water treatment.

• Ocean and Polar Research
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• v.28 no.1
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• pp.57-65
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• 2006

Various marine plankton were observed in the ballast water of vessels entering Incheon and Busan harbors. The ballast water of which age ranged from 2 to 54 days originated from the coastal waters of New Zealand, Taiwan, Singapore, Japan, Hong Kong and Pakistan, and from the Pacific Ocean. The total number of marine plankton taxa in 9 ballast tanks of different ships was 170: 90 phytoplankton, 24 protozoa and 56 zooplankton. The most diverse taxonomic groups were diatoms in phytoplankton, ciliates in protozoa and copepods in zooplankton. Classifying the specimens by size, above 50% of the number species of phytoplankton belonged to the size range between 50 and 150 Un. Protozoa and metazooplankton were found frequently in the size range between 50 and $120{\mu}m$ and 500 and $1,000{\mu}m$, respectively. The relationship between the species number and the age of ballast water was not significant. This is because of difference of filtration amounts derived from discordance of collecting samples. Among plankton observed in ballast water, some harmful algae and non-indigenous aquatic species were identified. Therefore, we need to investigate whether these species can inhabit in Korean coastal waters in further study.

• Journal of Korean Society on Water Environment
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• v.26 no.4
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• pp.614-621
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• 2010

A indigenous freshwater bivalve Unio douglasiae was introduced to compare the differences in the efficacy of algal bloom control and the appearances of mussel-mediated adverse effects between two different reservoirs such as mesotrophic (Shingu r.) and hypertrophic (Seokmun r.). We constructed the study mesocosm in the shore of each reservoir, stocked the mussel at density of $30indiv./m^3$ for 7 days, and measured daily the phytoplankton density and water quality. In mesotrophic reservoir, even though approximately 38% of suspended solids and chlorophyll-a was reduced by stocked bivalves for the first 3 days, algal density, ammonia and soluble reactive phosphorus gradually increased with increasing mussel death. In hypertrophic reservoir, mussels strongly inhibited suspended solids and chlorophyll-a by the termination of study with no increase of mussel death and nutrient, especially ammonia concentration. In both reservoirs, a strong selectivity showed mussels preferred to diatom rather than cyanobacteria and green algae without algal density and nutrient level. Our results indicate that an introduction of freshwater bivalve U. douglasiae is more strategic to improve water quality of hypertrophic than mesotrophic reservoir, but many preliminary studies on the treatment method and the selection of target water system are required.

• 한국해양학회지
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• v.28 no.1
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• pp.52-68
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• 1993

Effects of iron and/or chelator addition on primary production in the equatorial Upwelling system were studied during the TOGA(Tropical oceans and Global Atmosphere) and EPOCS (Equatorial Pacific ocean Climate Studies) cruises in June and November-December of 1989. Changes in the phytoplankton biomass and the degree of iron stress were estimated using the changes in vivo fluorescence before and after the addition of DCMU, which is an inhibitor of photosynthetic electron transposer system. Nitrate uptake was measured using /SUP 45/N labeled KNO$_3$ to estimate the new production. When samples were taken from the Upwelling area where nitrate concentration was higher than 5 uM, there were significant differences between the control and cheated iron treatments in vivo fluorescence and in nitrate uptake capacity. However, CFC (Cellular fluorescence capacity) did not show any significant difference between the control and treatments until nutrient limitation becomes severse and cells become shifted-down. Outside of the Upwelling area where surface nitrate concentration was low (below 0.5 uM), there was no significant difference between the control and treatments in vivo fluorescence and CFC. It is evident that primary and new production in the equatorial Pacific Upwelling region are limited by the availability of iron. However, the physiology of phytoplankton indigenous to this region does not appear to be iron stressed judging from CFC values.

• Korean Journal of Ecology and Environment
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• v.44 no.1
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• pp.31-41
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• 2011

To optimize the natural chemical agents against nuisance phytoplankton, we examined algal removal activity (ABA) of Plant-Mineral Composite (PMC), which already developed by our teams (Kim et al., 2010), on various conditions. The PMC are consisted of extracted-mixtures with indigenous plants (Camellia sinensis, Quercusacutissima and Castanea crenata) and minerals (Loess, Quartz porphyry, and natural zeolite), and characterized by coagulation and floating of low-density suspended solids. A simple extraction process was adopted, such as drying and grinding of raw material, water-extraction by high temperature-sonication and filtering. All tests were performed in 3 L plastic chambers varying conditions; six different concentrations ($0{\sim}1.0\;mL\;L^{-1}$), six light intensities ($8{\sim}1,400\;{\mu}mol\;m^{-2}s^{-1}$), three temperatures ($10{\sim}30^{\circ}C$), four pHs (7~10), five water depths (10~50 cm), and three different waters dominated by cyanobacteria, diatom, and green algae, respectively. Results indicate that the highest ABA of PMC was seen at $0.05\;mL\;L^{-1}$ in treatment concentrations, where showed a reduction of more than 80% of control phytoplankton biomass, while $1,400\;{\mu}mol\;m^{-2}s^{-1}$ in light intensity (>90%), $20{\sim}30^{\circ}C$ temperature (>60%), 7~9 in pH (>90%), below 50 cm in water depth (>90%), and cyanobacterial dominating waters (>80%), respectively. Over the test, ABA of PMC were more obvious on the algal biomass (chlorophyll-${\alpha}$) than suspended solids, suggesting a selectivity of PMC to particle size or natures. These results suggest that PMC agents can play an important role as natural agents to remove the nuisant algal aggregates or seston of eutrophic lake, where occur cyanobacterial bloom in a shallow shore of lake during warm season.

• Korean Journal of Fisheries and Aquatic Sciences
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• v.15 no.1
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• pp.1-25
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• 1982

Limnological study on the physico-chemical properties and biological characteristics of the Lake Ok-Jeong was made from May 1980 to August 1981. For the planktonic organisms in the lake, species composition, seasonal change and diurnal vertical distribution based on the monthly plankton samples were investigated in conjunction with the physico-chemical properties of the body of water in the lake. Analysis of temperature revealed that there were three distinctive periods in terms of vertical mixing of the water column. During the winter season (November-March) the vertical column was completely mixed, and no temperature gradient was observed. In February temperature of the whole column from the surface to the bottom was $3.5^{\circ}C$, which was the minimum value. With seasonal warming in spring, surface water forms thermoclines at the depth of 0-10 m from April to June. In summer (July-October) the surface mixing layer was deepened to form a strong thermocline at the depth of 15-25 m. At this time surface water reached up to $28.2^{\circ}C$ in August, accompanied by a significant increase in the temperature of bottom layer. Maximum bottom temperature was $r5^{\circ}C$ which occurred in September, thus showing that this lake keeps a significant turbulence Aehgh the hypolimnial layer. As autumn cooling proceeded summer stratification was destroyed from the end of October resulting in vertical mixing. In surface layer seasonal changes of pH were within the range from 6.8 in January to 9.0 in guutuost. Thighest value observed in August was mainly due to the photosynthetic activity of the phytoplankton. In the surface layer DO was always saturated throughout the year. Particularly in winter (January-April) the surface water was oversaturated (Max. 15.2 ppm in March). Vertical variation of DO was not remarkable, and bottom water was fairly well oxygenated. Transparency was closely related to the phytoplankton bloom. The highest value (4.6 m) was recorded in February when the primary production was low. During summer transparency decreased hand the lowest value (0.9 m) was recorded in August. It is mainly due to the dense blooming of gnabaena spiroides var. crassa in the surface layer. A. The amount of inorganic matters (Ca, Mg, Fe) reveals that Lake Ok-Jeong is classified as a soft-water lake. The amount of Cl, $NO_3-N$ and COD in 1981 was slightly higher than those in 1980. Heavy metals (Zn, Cu, Pb, Cd and Hg) were not detectable throughout the study period. During the study period 107 species of planktonic organisms representing 72 genera were identified. They include 12 species of Cyanophyta, 19 species of Bacillariophyta, 23 species of Chlorophyta, 14 species of Protozoa, 29 species of Rotifera, 4 species of Cladocera and 6 species of Copepoda. Bimodal blooming of phytoplankton was observed. A large blooming ($1,504\times10^3\;cells/l$ in October) was observed from July to October; a small blooming was present ($236\times10^3\;cells/l$ in February) from January to April. The dominant phytoplankton species include Melosira granulata, Anabaena spiroides, Asterionella gracillima and Microcystis aeruginota, which were classified into three seasonal groups : summer group, winter group and the whole year group. The sumner group includes Melosira granulate and Anabaena spiroides ; the winter group includes Asterionella gracillima and Synedra acus, S. ulna: the whole year group includes Microtystis aeruginosa and Ankistrodesmus falcatus. It is noted that M. granulate tends to aggregate in the bottom layer from January to August. The dominant zooplankters were Thermocpclops taihokuensis, Difflugia corona, Bosmina longirostris, Bosminopsis deitersi, Keratelle quadrata and Asplanchna priodonta. A single peak of zooplankton growth was observed and maximum zooplankton occurrence was present in July. Diurnal vertical migration was revealed by Microcystis aeruginosa, M. incerta, Anabaena spiroides, Melosira granulata, and Bosmina longirostris. Of these, M. granulata descends to the bottom and forms aggregation after sunset. B. longirostris shows fairly typical nocturnal migration. They ascends to the surface after sunset and disperse in the whole water column during night. Foully one species of fish representing 31 genera were collected. Of these 13 species including Pseudoperilnmpus uyekii and Coreoleuciscus splendidus were indigenous species of Korean inland waters. The indicator species of water quality determination include Microcystis aeruginosa, Melosira granulata, Asterionelta gracillima, Brachionus calyciflorus, Filinia longiseta, Conochiloides natans, Asplanchna priodonta, Difflugia corona, Eudorina elegans, Ceratium hirundinella, Bosmina longirostris, Bosminopsis deitersi, Heliodiaptomus kikuchii and Thermocyclops taihokuensis. These species have been known the indicator groups which are commonly found in the eutrophic lakes. Based on these planktonic indicators Lake Ok-Jeong can be classified into an eutrophic lake.

• Korean Journal of Ecology and Environment
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• v.43 no.3
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• pp.437-450
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• 2010

Ecological influences of indigenous freshwater bivalve Unio douglasiae on the water quality and epilithic diatom community was examined with artificial stream (AST), which constructed in a laboratory daily receiving the eutrophic lake water. For the colonization of new periphyton community, forty commercial slide glasses were deposited as a substrate into the lowest part of each AST. Prior to 1 week, the AST was operated to induce the freely-colonization of the algal community in the absence of mussels. After the mussels was introduced at 435 indiv. $m^{-2}$ between step 1 and step 5, the passed water and substrates were daily collected to analysis the change of water quality and lotic and lentic algae abundance for 10 days. Compared to the control, turbidity (60.0% of control), suspended solids (62.5%), and chlorophyll-$\alpha$ (72.2%) in mussel-passed waters were decreased significantly, while a strong increase of ammonia (up to 800% of control) was companied with the decrease of dissolved oxygen (19.5% of control) and total phosphorus (23.9%), respectively. On average, the concentrations of suspended solids (67.0% of control) and chlorophyll-$\alpha$ (89.4%) in mussel-treated substrates were remarkably increased, however algal abundance in its water simultaneously decreased. These results indicate that incidentally or purposely mussel introductions can decrease organic matter of the stream and increase transparency of stream water, however, mussel-mediated nutrient and pseudofeces release may stimulate the adverse growth of periphyton or phytoplankton community in the lower stream or reservoir.

• The Sea:JOURNAL OF THE KOREAN SOCIETY OF OCEANOGRAPHY
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• v.6 no.3
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• pp.201-210
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• 2001

Salinity, DIN, DIP, DIN/DIP and indigenous algal assay were determined to estimate the limiting nutrient for phytoplankton growth in Gwangyang Bay, South Sea of Korea. Seawater samples were collected at surface and bot-tom water in 4 November 1999 (dry season) and 2 September 2000 (after heavy rain). In 4 November 1999, the salinity, DIN, DIP and DIN/DIP were 29.92 psu, 13.59 ${\mu}M$, 3.41 ${\mu}M$ and 4.14 respectively. In 2 September 2000, These values were 24.62 psu, 27.77 ${\mu}M$, 2.82 ${\mu}M$ and 9.79 respectively. The DIN and DIP concentrations in this study were higher than Deukryang, Yeoja and Gamak Bay, South Sea of Korea. Especially, DIP concentration was 8 times high compared to Deutryang, Yeoja and Gamak Bay. The main sources of nitrogen seem to be freshwater runoff from Somjin River and industrial wastewater. But, the main sources of phosphorus seem to be industrial wastewater around Gwangyang Bay. The limiting nutrient was nitrogen at all station in 4 November 1999. The limiting nutrient was also nitrogen in 2 September 2000 in spite of heavy rain observed because of relatively much volume of phosphorus sup-plied from point sources than nitrogen. In case of below 20 psu in salinity by heavy rain, the limiting nutrient willbe shift from nitrogen to phosphorus at some area of Somjin River estuary. But the limiting nutrient will be never shift to phosphorus throughout Gwangyang Bay, eastern coast of Yeoja and Dolsan because of much volume of phosphorus runoff from point source in coastal area of Gwangyang Bay.