• Journal of Environmental Science International
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• v.18 no.4
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• pp.435-443
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• 2009

A bacterial strain SY-97 that showed algicidal activity against Cochlodinium polykrikoides was isolated from coastal water of Uljin (eastern coast of Korea) in August, 2005. The isolated strain was identified as Brachybacterium sp. by morphological and biological tests, and analysis of 16S rDNA sequence. The optimal culture conditions for the growth of strain SY-97 were $30^{\circ}C$, initial pH 7.0, and salinity 2.0%. From the result of cell culture insert experiment, Brachybacterium sp. SY-97 is assumed to produce secondary metabolites which have algicidal activity. When 10% culture filtrate of this strain was applied to C. polykrikoides ($1.2{\times}10^4\;cells/m{\ell}$) cultures, 100% of C. polykrikoides cells was destroyed within 15 hours. The released algicides were heat-tolerant to $100^{\circ}C$ and stable in pH $6.0{\sim}10.0$. These results suggest that Brachybacterium sp. SY-97 is potentially useful for controlling outbreaks of C. polykrikoides.

• Journal of the Korean Society for Marine Environment & Energy
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• v.14 no.1
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• pp.19-31
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• 2011

This study has examined the water quality environment of six areas in the mid-southern coastal waters of Korea in order to find the significance between water quality and algal blooms of the area, based on the last 17 years of data offered by the National Fisheries Research and Development Institute. Water temperature in these areas fluctuated with a three to five year of period, and revealed little yearly variations at the surface layer while slowly decreasing at the bottom layer. On the other hand, salinity tended to increase both the surface and bottom layers. Algal blooms had a tendency to decrease in their outbreaks and causative species, with a peak of the middle of 1990s. C. polykrikoides prevailed in the entire areas, and in particular, almost annually appeared in Goheung coastal area since 1995. C. polykrikoides blooms occurred when a mean water temperature was approximately $26^{\circ}C$, and salinities were between 31.00 and 31.50 but exceptionally 28.68 in Yeosu Coast. However, the concentrations of DIN, DIP, TN, TP, including DO, turned out not to be such significant factors for the outbreaks of C. polykrikoides blooms. Therefore, water temperature was judged as the most controlling factor for the outbreak of C. polykrikoides blooms.

• ALGAE
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• v.31 no.4
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• pp.373-378
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• 2016

Cochlodinium polykrikoides is a red-tide forming dinoflagellate that causes significant worldwide impacts on aquaculture industries and the marine ecosystem. There have been extensive studies on managing and preventing C. polykrikoides blooms, but it has been difficult to identify an effective method to control the bloom development. There is also limited genome information on the molecular mechanisms involved in its various ecophysiology and metabolism processes. Thus, comprehensive genome information is required to better understand harmful algal blooms caused by C. polykrikoides. We estimated the C. polykrikoides genome size using flow cytometry, with detection of the fluorescence of DNA stained with propidium iodide (PI). The nuclear genome size of C. polykrikoides was 100.97 Gb, as calculated by comparing its mean fluorescence intensity (MFI) to the MFI of Mus musculus, which is 2.8 Gb. The exceptionally large genome size of C. polykrikoides might indicate its complex physiological and metabolic characteristics. Our optimized protocol for estimating the nuclear genome size of a dinoflagellate using flow cytometry with PI can be applied in studies of other marine organisms.

• Proceedings of KOSOMES biannual meeting
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• 2007.11a
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• pp.187-188
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• 2007

The fluctuations of biochemical and molecular activities III the harmful dinoflagellate, Cochlodinium polykrikoides, depending on water temperatures, were studied. In genomic DNA concentration, a similar value of 0.6 was shown at $12^{\circ}C$ and $15^{\circ}C$, but significantly increasing DNA from $18^{\circ}C$ (p<0.05), with a maximum of 1.8 at $24^{\circ}C$. After$24^{\circ}C$, the DNA significantly decreased to 0.6. Likely, the concentrations of RNA and total protein were at their highest values of 1.7 and 0.07 g $mL^1$ at $24^{\circ}C$, respectively. In contrast to ONA, RNA and total protein began to increase at $15^{\circ}C$. Oxygen availability between lower and higher temperatures was significantly different and increased from $18^{\circ}C$ according to light intensity, regardless of wavelengths (p<0.05). At $24^{\circ}C$, the highest value of the maximum electron transport rate (ETRmax), ranging from 537.9 (Ch 1) to 602.5 mol electrons $g^{-1}$ Ch1 a $s^{-1}$ (Ch 4), was also shown. Nitrate reductase (NR) and ATPase activities were at their highest values of 0.11 mol $NO_2^-g^{-1}$ Ch1 a $h^{-1}$ and 0.78 pmol 100 $mg^{-1}$ $at^2$ $4^{\circ}C$, respectively. When the cells cultured at $15^{\circ}C$, NR and ATPase activities significantly increased compared to $12^{\circ}C$ (p<0.05). In an analysis of CHN, the concentration of C and N also significantly increased (p<0.05). However, at $27^{\circ}C$, most of the molecular and biochemical movements were much lower, compared to $24^{\circ}C$. These results suggest that C. polykrikoides is very sensitive biochemical and molecular activities depending on water temperatures. Possibly, it is desirable to estimate at $18^{\circ}C$ the initiation of the massive blooming development of C. polykrikoides. In nature, it will be very difficult to maintain the massive blooms after $24^{\circ}C$ because of the possibility of significantly decreasing the molecular movement and activity of C. polykrikoides.

• The Sea:JOURNAL OF THE KOREAN SOCIETY OF OCEANOGRAPHY
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• v.21 no.1
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• pp.44-47
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• 2016

Real-time PCR was applied to early warning of red tide. For early warning of red tide at $10cells\;mL^{-1}$, Cochlodinium polykrikoides specific real-time PCR was used as a complement of microscopy that has a lower detection sensitivity. C. polykrikoides appeared extensively in Tongyeong, Namhae waters at low densities in the end of June, and early warning of C. polyrkrikoides blooms was issued on 2 August 2015.

• Fisheries and Aquatic Sciences
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• v.6 no.3
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• pp.160-163
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• 2003

Mortality of several species of fish and shellfish exposed to Harmful Algal Blooms (HABs) caused by Cochlodinium polykrikoides, Heterosigam akashiwo, Alexandrium tamarense, Eutreptiella gymnastica, Heterocapsa triquetra and Prorocentrum micans was studied. When fish were exposed to a cell density of 8,000 cells $mL^{-1}$ in C. polykrikoides, $35\%$ of flatfish and darkbanded rockfish died within 48 hrs. However, jacopever rockfish had mortality of higher than $85\%$. Rock bream, filefish and red sea bream showed $100\%$ mortality within 10 hrs with an exposure cell density of 8,000 cells $mL^{-1}$. The rest of HABs except for C. polykrikoides showed that there was no fish and shellfish death throughout the 48 hrs even in the maximum cell density of 100,000 cells $mL^{-1}$ These results imply that C. polykrikoides can have a serious impact on fish mortality and it is regarded as an ichthyotoxic dinoflagellate. The fish death may be attributed to anoxia caused by a combination of the production of reactive oxygen species (ROS) and polysaccharide from C. polykrikoides during blooms.

• Journal of Environmental Science International
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• v.17 no.4
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• pp.439-449
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• 2008

Algicidal bacterium was isolated from sea water during the declining period of Cochlodinium polykrikoides blooms and this bacterium had a significant algicidal activity against C. polykrikoides. In this study, algicidal bacterium was identified on the basis of biochemical and chemotaxonomic characteristics, and analysis of 16S rDNA sequences. The algicidal bacterium showed 98.6% homology with Micrococcus luteus ATCC $381^T$. Therefore, this bacterium was designated Micrococcus luteus SY-13. The optimal culture conditions of the algicidal bacterium was $25^{\circ}C$, initial pH 8.0, and 3.0% NaCl concentration. M. luteus SY-13 is assumed to produce secondary metabolites which have algicidal activity. When 10% culture filtrate of this strain was applied to C. polykrikoides ($1.0\;{\times}\;10^4\;cells/ml$) cultures, over 98% of C, polykrikoides cells were destroyed within 6 hours. The culture filtrate of M. luteus SY-13 exhibited similar algicidal activity after heat-treatment at $121^{\circ}C$ for 15 min. While algicidal activity remained in filtrates with pH adjusted to 8.0, loss of algicidal activity occurred when the pHs of filtrates were adjusted to over 9.0 or heat-treated at $121{\times}180^{\circ}C$ for 1 hour. M. luteus SY-13 showed significant algicidal activities against C. polykrikoides (98.9%) and a wide algicidal range against various harmful algal bloom (HAB) species. However, there was no algicidal effect on diatom and marine livefood organisms except Isocrysis galbana. These results suggest that M. luteus SY-13 could be a candidate for use in the control of HABs.

• Journal of the Korean Society for Marine Environment & Energy
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• v.18 no.3
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• pp.166-178
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• 2015

We investigated marine environmental characteristics of Goheung coastal areas in August where is known to be the first outbreak site of Cochlodinium polykrikoides (hereafter C. polykrikoides) blooms, based on the oceanographic data observed from 1993 to 2013 around the Korean southern coastal waters including Eastern China Sea by National Fisheries Research and Development Institute (NFRDI). The data of NOAA/NGSST satellite images as well as numerical simulation results by Seo et al. [2013] were also used for analysis. Water temperatures at the surface and bottom layers in Goheung coast, i.e. Narodo, were $25.0^{\circ}C$ and $23.7^{\circ}C$ so that they were higher than $23.8^{\circ}C$ and $19.4^{\circ}C$ in Geoje coast where is a reference site, respectively. In addition, salinities at the surface and bottom layers in Goheung coast were 31.78 psu and 31.98 psu so that they were a little higher than 31.54 psu at the surface but a little lower than 32.79 psu at the bottom in Geoje coast, respectively. That is, the differences in water temperature or salinity between the surface and bottom layers in Goheung coast in August were not large compared to Geoje coast. This suggests that stratification in Goheung coast in August is fairly weak or may not be established. In addition, the concentrations of DIN and DIP at the surface layer were 0.068 mg/L ($4.86{\mu}M$) and 0.015 mg/L ($5.14{\mu}M$) in Goheung coast while 0.072 mg/L ($5.14{\mu}M$) and 0.01 mg/L ($0.32{\mu}M$) in Geoje coast, so they did not indicate a meaningful difference. On the other hand, when C. polykrikoides blooms, water temperature and salinity in August at the station 317-22 ($31.5^{\circ}N$, $124^{\circ}E$) of the East China Sea, where is near the mouth of Yangtze River, were $27.8^{\circ}C$ and 31.61 psu, respectively. Thus, water temperature was much higher whereas salinity was almost similar compared to Goheung coast. Furthermore, concentrations of $NO_3-N$ and $PO_4-P$ in the East China Sea in August were remarkably high compared to Goheung coast. When C. polykrikoides blooms, according to not only the image data of satellites NOAA/NGSST but also numerical experiment results by Seo et al.[2013], the freshwater out of Yangtze River was judged to clearly affect the Korean southern coastal waters. Therefore, the supply of nutrients in terms of Yangtze River may greatly contribute to the outbreak of C. polykrikoides blooms in Goheung coast in summer.

• Journal of Life Science
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• v.15 no.3 s.70
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• pp.468-473
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• 2005

The coastal regions of Yeosu, the South Sea of Korea, has occurred annually the red tide which is caused by potentially ichthyotoxic dinoflagellate C. polykrikoides, with a wide avenue for exchange with oceanic waters and freshwater runoff from Sumjin river. We attempted to examine the variability in response to vertical migration and concentration of extracted DNA/RNA of C. polykrikoides exposed to salinity-stratified waters. The experimental aquarium of the 60 liter was employed to culture C. polykrikoides. One aquarium was supplied with only sea water, the other was consisted of sea water and freshwater. Experiment was conducted for 5 days. In experimental column (mixture of freshwater and sea water), salinity was maintained to 20 at upper and approximately 30 at bottom during the period of this study. The fluctuation with related to dissolved oxygen and pH was similar pattern to both columns. Chlorophyll a was significantly higher value at upper than bottom. During 24h, chlorphyll a on experimental column was extremely high on the top as soon as lighting, compared with control. With elapsed time, the gap between experimental and control columns was a little. In darkness, chlorophyll a was not significantly different between upper and bottom, most cells appeared to randomly distribute on column regardless of water layer. Fluctuation with related to concentration of extracted DNA and RNA based on ratio of absorbance of 260 and 280 nm in experimental column was higher at final day or diel migration than control. These results implied that a large volume of freshwater could be associated with influence of concentration of DNA and RNA, in particular, rapid upward movement caused massive fish kills as soon as sunset.

• The Sea:JOURNAL OF THE KOREAN SOCIETY OF OCEANOGRAPHY
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• v.19 no.4
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• pp.232-242
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• 2014

We investigated the interspecific competition between the harmful dinoflagellate Cochlodinium polykrikoides and diatom Skeletonema sp. based on the utilization and uptake of dissolved organic nutrients. C. polykrikoides and S. costatum were able to grow using dissolved organic nitrogen (DON) and dissolved organic phosphorus (DOP) as well as dissolved inorganic nitrogen (DIN) and dissolved inorganic phosphorus (DIP). This result indicates that the utilization of dissolved organic nutrients may play a role in surviving strategy in the DIN or DIP-limited environments. The half-saturation constants (Ks) of urea and glycerophosphate (glycero-P) calculated from uptake kinetics experiment of C. polykrikoides was lower than those of Skeletonema sp. This result indicates that Skeletonema sp. have higher affinity for dissolved organic nutrients, such as urea and glycero-P, than C. polykrikoides. Although Skeletonema sp. have higher affinity of dissolved organic nutrients, C. polykrikoides could effectively uptake for urea and glycero-P at sub-saturating nutrient concentrations (${\alpha}$ (${\rho}_{max}/Ks$) of C. polykrikoides was higher than Skeletonema sp.. Therefore, C. polykrikoides by utilization and effectively uptake of dissolved organic nutrients under monoculture may have an advantageous position in the interspecific competition with Skeletonema sp. in the low nutrient environments.