• Korean Journal of Fisheries and Aquatic Sciences
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• v.34 no.5
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• pp.445-456
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• 2001

In the 1990s, Cochlodinium polykikoides red tide has been annually occurred in the southern coast of Korea and caused the mass damage to the fisheries with a huge amount of economic loss. The present study was done to establish the biological foundation for the elucidation of the mechanism of C. polykikoiaes red tide. The growth response of C. polykikoides to physico-chemical factors such as temperature, salinity, pH, and light intensity were examined using axenic cultures to evaluate the relative importance of these factors on the dynamics of natural populations, It was found that the highest growth conditions were $25^{\circ}C,\;40\%_{\circ}$, pH 7.5, and 7,500 lux, respectively. The tolerable salinity range of growth was relatively wide at an optimum temperature and was reduced to a much narrower range at a sub-optimum temperature. These findings indicate that C. polykikoides is an eurythermal and euryhaline organism. The organism demanded higher light intensity and oceanic pH narrow in its growth. C. polykikoides utilize inorganic nutrients, such as nitrate and ammonium as N, and phosphate as P. The nutritional requirements of C. polykikoides were $40{\mu}M$ for nitrate, $50{\mu}M$ for ammonium, and $5{\mu}M$ for phosphate. The half saturation constant (Ks) for growth was $2.10{\mu}M$ for nitrate, $1.03{\mu}M$ for ammonium, and $0.57{\mu}M$ for phosphate. These values were comparatively smaller than those of other dinoflagellates reported previously. We confirmed that the organism is characterized as an eutrophic species. However, ammonium Ks value is smaller than that of other eutrophic species, This result indicates that C. polykikoides red tide may outbreak in the waters which eutrophication is in progress rather than eutrophicated waters. C. polykikoides preferred ammonium better than nitrate as a nitrogen source when in a growth stage, Therefore, our results indicate that ammonium is more important nutrient on the growth of the organism in comparison with other inorganic nutrients and C. polykikoides red tide is related with the increased ammonium concentration in the coastal waters.

• Korean Journal of Fisheries and Aquatic Sciences
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• v.35 no.6
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• pp.568-577
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• 2002

The spreading Cocuoainim polykikoides bloom in the southern coastal waters of Korea was simulated using numerical model including the physical processes of water flow and the chemical processes of increasing cell of C. polykikoides by uptake of dissolved nutrients. The circulation of sea water was simulated by two dimensional tide model reflecting the main four tidal components of $M_2,\;S_2,\;K_1,\;O_1$, and permanent current was driven by inflow/outflow across open boundaries. According to the result of model which tidal and permanent current were reflected simultaneously, eastward flows entering the southern waters from the western waters of Korea are dominant but westward flows are weak relatively. These result suggest that it is difficult for initial C. polykikoides bloom generated in the coastal waters of Goheung to move to the western coast of Korea through Jeju Strait. For spreading model of C. poiyhikoides, the range of generating distribution and the generating time of C. polykikoides bloom in coastal area are similar to those of observation data in the field. Wind is the most important factor in moving and distribution of red tide. Permanent current flowing eastward is also considered to be important factor and tidal current was a little influenced.

• Korean Journal of Fisheries and Aquatic Sciences
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• v.31 no.5
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• pp.767-773
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• 1998

In this study, we have investigated the distributions and killing effects of marine bacteria that tend to kill the red tide microalgae, C. polykikoides in the area of Masan bay from June to October, 1996. To summarize, C. polykikoides killing bacteria were detected at $10^2$ to $10^3$ cells/ml of seawater samples during the survey period, and the bloom was observed in September by containing $4.8\times10^3$cells/ml. It appears however that the number of these bacteria is decreased ($2.0\times10^2$cells/ml) in October, A total of 110 strains were isolated from seawater samples and seawater filtrate (pore size, 0.8 $\mu$m)-containing mixed culture of C. polykikoides in which the mixed culture was grown in f/2 medium. As results we have successfully isolated Micrococcus sp. LG-1 which decreased to less than 10cells/ml within 6days and 5days sfter inoculation of Micrococcus sp. LG-1 into the la9 and logarithmic growth phases of C. polykrikoides respectively. Therefore, it appears that inoculation of Micrococcus sp. LG-1 against the logarithmic C. polykrikoides is more effective than the lag growth phase, (n addition, the killing effects were increased in accordance with bacterial cell densities inoculated in a dose dependent manner. Especially, the filtrate of kitling bacterium culture (nore size, 0.2 $\mu$m) revealed a dramatic effect in which C. polykrikoides were decreased to less than 10 cells/mf of culture within 1 hr, 1,5 hrs, 1,5 hrs, 3.5 hrs. and 5,5 hrs after inoculations of the culture filtrate with concentration of $30\%,\;20\%,\;10\%,\;5\%$ and $2.5\%$, respectively. Moreover Micrococcus sp. LG-1 showed a selective specificity against C. polykrikoides and any other killing effects of Micrococcus sp. LG-1 were not observed against Alexandrium tamarense, Prorocentrum micans, Scrippsiella trochoidea. ana Gymnodinium sanguineum.

• Korean Journal of Fisheries and Aquatic Sciences
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• v.34 no.5
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• pp.457-464
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• 2001

Algal growth potential (AGP) assay using Cochlodinium polykrikoides was conducted in Saryang Island coast where C. polykrikoides red tide occurred annually from July to October 1998. The effects of macro- and micro-nutrients on the growth of C. polykrikoides were specifically evaluated by the algal assay method. Two different types of growth response of C. polykikoides for the addition of nutrients were clearly obseued. For both before and after C. polykrikoides occurrence, the growth of C. polykikoides was significantly stimulated by the addition of either nitrate or ammonium of $50{\mu]M$ with phosphate of $5{\mu}M$. The addition of a single nutrient had no clear effect on the growth of C. polyhikoides and the addition of trace metals, vitamins, and EDTA etc. did not stimulate the algal growth, also. This result indicates that both N and P potentially limited the growth of C. polyhikoides in this period. However, during a bloom of C. polyhikoides, the growth was unlikely to be stimulated by the addition of both macro- and micro-nutrients. At that time the nutrient concentration of Saryang Island coast was $24.33{\mu}M$ for ammonium, $1.61{\mu}M$ for phosphate, and $0.58{\mu}M$ for nitrate, respectively. The concentrations of nutrients increased, on average, 8.2-fold for ammonium and 4.8-fold for phosphate, decreased 3.3-fold for nitrate compared to both before and after the red tide. This result shows that the growth of C. polykikoides was not limited by the nutrients during the bloom in September. Therefore, our results suggest that the C. polykrikoides red tide may outbreak especially when the water is fertilized due to the increased N and P.

• Korean Journal of Fisheries and Aquatic Sciences
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• v.33 no.4
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• pp.331-338
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• 2000

The algicidal effects of marine bacteria were investigated and a strain, which had the strongest algicidal activity against the harmful dinoflagellate, Cochiodinim polykrikoides was selected. The bacterium was isolated in seawater during the period of blooming of C. polykrikoides in Masan Bay. This algicidal bacterium was identified as Micrococcus sp. LG-5 by means of morphological and biochemical tests. The optimal culture conditions of Micrococcus sp, LG-5 were $25^{\circ}C,\;pH 7.0\;and\;3.0{\%}$ NaCl concentration. The algicidal activity of Micrococcus sp. LG-5 was significantly increased to maximum value in the late of logarithmic phase of cell cuture. In addition, the culture filtrate ($pore size,\;0.1{\mu}m$) of Microcoocus sp. LG-5 showed strong algicidal effects. The cell numbers of C. polykikoides were decreased from $1.2{\times}10^4 cells/ml\;to\;less\;than\;2{\times}10^3\;cells/ml$ within 3, 6, 30 hours at the concentrations of culture filtrate $10{\%},\;5{\%}\;and\;1{\%}$, respectively. These results indicated that the algicidal effect was mediated by certain substances released from Microooccus sp. LG-5.

• Korean Journal of Fisheries and Aquatic Sciences
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• v.34 no.5
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• pp.536-544
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• 2001

Three harmful algal bloom species with similar morphology, Cochlodinium polykrikoides, Gyodinium impudicum and Gymodinium catenatum have damaged to aquatic animals or human health by either making massive blooms or intoxication of shellfishes in a food chain. Eco-physiological and hydrodynamic studies on the harmful algae offer useful informations in the understanding their bloom mechanism by giving promising data for the prediction and modelling of harmful algal blooms event. Thus, we studied the abundance of these species in the coastal area of South Sea of Korea and their effects of temperature, salinity, irradiance and nutrient on the growth for the isolates. The timing for initial appearance of the three species around the coastal area of Namhaedo, Narodo and Wando was between Bate July and late August in 1999 when water temperature ranged from $22.8^{\circ}C\;to\;26.5^{\circ}C$ Vegetative cells of C. polykrikoides and G. impudicum were abundant until late September when water temperature had been dropped to less than $23^{\circ}C$. By contrast, vegetative cell of G. catenatum disappeared before early September, showing shorter period of abundance than the other two species in the South Sea. Both G. impudicum and G. catenatum revealed comparatively low density with a maximal cell density of 3,460 cells/L and 440 cells/L, respectively without making any bloom, while C. polykrikoides made massive blooms with a maximal cell density more than $40\times10^6$cells/L, The three species showed a better growth at the relatively higher water temperature ranging from 22 to $28^{\circ}C$ with their maximal growth rate at $25^{\circ}C$ in culture, which almost corresponded with the water temperature during the outbreak of C. polykrikoides in the coastal area of South Sea. Also, they all showed a relatively higher growth at the salinity from 30 to $35\%$. Specially, G. impudicum showed the euryhalic characteristics among the species, On the other hand, growth rate of G. catenatum decreased sharply with the increase of water temperature at the experimental ranges more than $35\%$. The higher of light intensities showed the better growth rates for the three species, Moreover, C. polykrikoides and G. impudirum continued their exponential growth even at 7,500 lux, the highest level of light intensity in the experiment, Therefore, It is assumed that C. polykrikoides has a physiological capability to adapt and utilize higher irradiance resulting in the higher growth rate without any photo inhibition response at the sea surface where there is usually strong irradiance during its blooming season. Although C. poiykikoides and G. impudicum continued their linear growth with the increase of nitrate ($NO_3^-$) and ammonium ($NH_4^-$) concentrations at less than the $40{\mu}M$, they didn't show any significant differences in growth rates with the increase of nitrate and ammonium concentrations at more than $40{\mu}M$, signifying that the nitrogen critical point for the growth of the two species stands between 13.5 and $40{\mu}M$. Also, even though both of the two species continued their linear growth with the increase of phosphate ($PO_4^{2-}$) concentrations at less than the $4.05{\mu}M$, there were no any significant differences in growth rates with the increase of phosphate concentrations at more than $4.05{\mu}M$, signifying that the phosphate critical point for the growth of the two species stands between 1.35 and $4.05{\mu}M$. On the other hand, C. polykrikoides has made blooms at the oligotrophic environment near Narodo and Namhaedo where the concentration of DIN and DIP are less than 1.2 and $0.3{\mu}M$, respectively. We attributed this phenomenon to its own ecological characteristics of diel vertical migration through which C. polykrikoides could uptake enough nutrients from the deep sea water near bottom during the night time irrespective of the lower nutrient pools in the surface water.

• Korean Journal of Fisheries and Aquatic Sciences
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• v.33 no.4
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• pp.339-347
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• 2000

An algicidal bacterium, Micrococcus sp. LG-5 against the harmful dinoflagellate, Cochlodinium polykrikoides was isolated. The optimal conditions for the highest algicidal activity of bacterial culture filtrate showed in the range of $20{\~}30^{\circ}C$, at pH 7.0 and $3.0{\%}$ of NaCl concentration. In addition, $IC_(50)(mean of 50{\%} inhibitory concentration)$ of the culture filtrate against C. polykrikoides after incubation of 5 days was $0.482{\%}$. To investigate heat and pH stability of the culture filtrate of Micrococcus sp. LG-5, the culture filtrate ($pore size, 0.1 {\mu}m$) was heated to $121^{\circ}C for 15 min$ and adjusted pH from 2.0 to 10.0. There were no significant changes in algicidal activity by heat treatment and the pH change between pH from 5.0 to 10.0. The algicidal substances produced from Micrococcus sp. LG-5 were mainly detected in the fraction of $10,000{\~}1,000$ MWCO (molecular weight cut-off). The culture filtrate of Micrococous sp. LG-5 showed antimicrobial activity against Enterococcus faecalis, Escheiichia coli, Uebsiella pneunioniae and Vibrio altinolyticus, but did not show against Pseudomonas aeminosa, P. Buorescens, Salmonella typhi, Staphylococcus aureus, V. cholerae and V parahaemolyicus. The culture filtrate of Micrococcus sp. LG-5 was examined against 16 phytoplankton species and showed the algicidal activity against Ajexandzium tuarense, Eutreptiella Drnnastin, Gymnodinium catenatum, G. mikimotoi, G. sanguineum, eyodinium impuaicum, Heterocapsa triquetra, Heterosipa akashiwo, Prorocentrum micans and Pyraminonas sp.. However no algicidal effects of Micrococcus sp. LG-5 were observed against Chlamydomonas sp., Cylindrotheoa closterium, P. mininum, P. triestimum, Pseudonieschia sp. and Sczipuiella trochoidea. On the other hand, algicidal activity on the tested marinelivefood was not detected except for Isochrysis galbana. In addition, physiological responses of cultured olive flounder (Paralichthys oliraceus) exposed to $1 and 10{\%}$ of the culture filtrate of Micrococcus sp. LG-5 were measured. There were no clear changes in AST, GGT, creatinine, urea, total cholesterol, total protein, albumine, $Mg^(+2), Ca^(+2), Na^+, K^+, and Cl^-$. These results indicate that olive flounders were not affected when they were exposed to the culture filtrate of Micrococcus sp. LG-5.