본 연구는 양식산 참돔과 넙치를 Cochlodinium polykrikoides 적조에 노출시켜 노출 시간에 따 른 생존율, 호흡수, 혈중 스트레스 지표 및 조직학적 변화를 비교 조사하였다. 대조구는 자연해 수를 사용하였고, 실험구는 C. polykrikoides 밀도를 5,500±200 cells/ml로 설정하였다. 그 결과, 참돔은 적조 노출 1시간 이내, 넙치는 적조 노출 5시간 이내 전량 폐사하였다. 생리학적 반응 을 분석한 결과, 참돔은 적조 노출 후 혈중 Glucose 농도가 감소하였으며, 혈중 GOT 및 GPT 농도는 증가하였고, 혈중 SOD 농도는 감소하고, CAT 및 GPx 농도는 증가하는 경향을 보였다. 넙치는 적조 노출 후 혈중 Cortisol 및 GOT, GPT 농도가 증가하였고, 혈중 Glucose 농도는 적조 노출 1시간째 증가한 이후 감소하였으며, 혈중 SOD, CAT, GPx 농도는 노출 1시간째 감소한 이후 증가하였다. 조직학적 분석 결과, 참돔과 넙치의 아가미에 구조적인 손상이 발생하였다. 결론적으로 5,500 cells/ml 밀도의 C. polykrikoides 적조 노출은 양식산 참돔과 넙치에게 산화 적 스트레스로 작용하여 체내 항산화 방어 기작을 활성화하고, 간과 아가미의 손상을 발생시 키는 것으로 나타났다.

In this study, the marine medaka Oryzias javanicus was exposed to two concentrations of non-toxin-producing red tide dinoflagellate C. polykrikoides (1,000 and 2,000 cells ml-1) for 96 h, and the time-course biochemical responses of antioxidant and immunity parameters were analyzed in the liver tissue. Significant ichthyotoxicity with increasing cell concentrations of C. polykrikoides and exposure period was observed for 96 h. Opercular respiratory rate was lowered in marine medaka exposed to 2,000 cells ml-1 of C. polykrikoides. Intracellular malondialdehyde (MDA) content significantly elevated in response to both cell concentrations. In the case of glutathione (GSH) content, the levels were significantly elevated by 1,000 cells ml-1 of C. polykrikoides, but the contents significantly depleted upon exposure to 2,000 cells ml-1 of C. polykrikoides. Similarly, enzymatic activities of catalase (CAT) and superoxide dismutase (SOD) were increased by 1,000 cells ml-1 of C. polykrikoides, whereas their activities were lowered by 2,000 cells ml-1 of C. polykrikoides. Analysis of the two immunity parameters, alternative complement pathway and lysozyme, showed significantly lowered activities in 2,000 cells ml-1 of C. polykrikoides-exposed liver tissue. These biochemical effects of C. polykrikoides on marine medaka would be helpful for understanding its acute effects in marine fish.

적조가 처음 시작되는 해역을 조기에 파악하기 위하여 Quantitative real-time PCR (qPCR)을 경남해역 적조현장에 활용하였다. 2019년 경남해역을 대상으로 Cochlodinium polykrikoides를 qPCR로 정량분석한 결과, 6월 초에 저밀도로(0.0015~0.0058 cells mL-1) 검출되기 시작하여 8월 중순에는 최대 0.163 cells mL-1 밀도로 증가하였고, 주로 남해도 주변에서 높게 검출되었다. 8월 말에는 현미경 검경으로 남해도 주변에서 높게 출현함이 확인되었고(최대 24 cells mL-1), 9월 2일에는 남해도에서 적조주의보가 발령되었고(최대 200 cells mL-1), 9월 11일에는 최대 12,000 cells mL-1까지 남해도 해역에서 발생하였다. 위 결과는 극미량의 C. polykrikoides이 적조발생 전에 남해도에서 검출 되었고 이후 같은 해역에서 적조가 발생되었음을 보여준다. 이는 qPCR이 극미량의 C. polykrikoides을 조기검출하는데 유용한 방법임을 보여준다.

Cochlodinium polykrikoides is a harmful algae that in Korea, has caused red tide in the southern and eastern coastal water of Korea from summer to early autumn since 1990, triggering fishing damage each year. To reduce and prevent this damage, loess is generally sprayed in the field conditions, but in recent years, the red tide coverage is becoming extensive, and thus there is a need to secure the removal materials for the long-term ocean environment changes. This study sought to confirm the use of kaolin with low trace metal content and the organism-extracted chitooligosaccharide as a C. polykrikoides removal material. The kaolin's removal efficiency by its concentration, using C. polykrikoides cultures, was measured, showing that there was no change to pH, and that, within 30 minutes, the removal effect stood at 21~62%. The test of chitooligosaccharide by its concentration showed that, with the pH change range (5.58~8.88), the removal effect was low at 9~30%. But, the successive injection of the mixture of kaolin and chitooligosaccharide into the cultures revealed that, while the kaolin concentration was irrelevant to the removal efficiency, upon the supply of over 2mL chitooligosaccharide, the removal efficiency increased to 62?91% with normal pH values (7.01~7.34) being measured. To ensure easy onsite spray of the kaolin and chitooligosaccharide mixture, its dry crashed substance was tested, revealing that, when the ratio of kaolin : chitooligosaccharide was 1:10, the removal efficiency was higher than 85%, and that there is a need for boosting cost-efficiency considering the easiness of spraying, drying time and the supply amount.

For the last few decades, especially during summer Korean aquaculture industry has been huge economic loss due to massive blooms caused by Cochlodinium polykrikoides. Moreover, high dispersion speed along the coast and high cell densities (above 10,000 cells ml−1) of C. polykrikoides resuling in red tides for a longer duration (at least for 30 days). C. polykrikoides can be used as a prolific source of extracellular sulfated polysaccharides. Sulfated polysaccharides have showed strong antiviral properties against influenza virus. However, little has been investigated about industrial application of sulfated polysaccharides as a high valuable byproduct from C. polykrikoides. Current commercial microalgal biomass production systems are costly and require advanced instrumental and cultural facilities. From economic point of natural blooms of C. polykrikoides, therefore it is likely to be utilized as a cost-effective way of microalgal biomass production for commercial applications.

Cochlodinium polykrikoides 적조는 매년 여름철에 남해안을 중심으로 발생하였으나, 2009년에는 적조가 전혀 발생되지 않았다. 본 연구는 남해안 여수부터 완도해역을 대상으로 조사정점 13개를 정하여 2007년부터 2009년까지 여름철 수질환경 및 식물플랑크톤 군집 동태를 적조가 발생된 연도(2007년과 2009년)와 적조가 발생되지 않은 연도(2009)년) 자료를 상호 비교하였다. 2009년 6월부터 남해안 집중강우로 인하여 7월의 평균수온이 20℃ 이하의 저수온과 8월에 염분이 31psu 이하의 저염분, pH가 8월에 8.4 정도를 보여 적조발생 년도에 비하여 해양환경의 큰 변화를 가져왔다. 또한 DIN 농도는 적조발생 년도에 비하여 약 2배 이상 높은 값을 보였고, N:P 및 Si:N 비율에서도 적조가 발생된 년도에 비하여 Redfield ratio가 16 이상을 나타내어 완전히 정반대의 양상을 보였다. 조사기간 동안의 대부분의 우점종으로는Chaetoceros spp., Skeletonema costatusm, Thalassiosira spp.와 같은 규조류가 대부분 출현되었고, 와편모조류로는 Ceratium spp. 등으로 나타났다. 특히 2009년 8월경에는 C. polykrikoides 대신에 Gonyaulax polygramma 종으로 적조를 형성하기도 하였다. 따라서 2009년 6월부터 집중강우 및 일조량 부족으로 인한 C. polykrikoides 대수증식기 시점에 수온, 염분 등과 같은 해양환경의 급격한 변화로 인하여 적조형성이 지연됨과 아울러 G. polygramma와 Chaetoceros 등과 같은 규조류의 세력에 밀려 2009년에는 C. polykrikoides 적조가 발생하지 않은 것으로 판단된다.

Ground seawater quality was investigated, and the algal growth potential (AGP) tests were performed along the Tongyeong coast to examine the inflow of materials, which is needed for the red tide species Cochlodinium polykrikoides to grow. The study was conducted to determine the mechanism for C. polykrikoidesred tides. Water temperature, salinity, pH, and dissolved oxygen (DO) ranged from 16.05 to 20.74°C, 18.20 to 32.11 psu, 6.00 to 7.61, and 3.41 to 7.91 mg/L (41.4– 96.1%), respectively. No seasonal variation was observed in water temperature. The salinity, pH, and DO saturations at most stations were lower than those of coastal seawater at Tongyeong. The NH4 +-N, NO2+3 --N, PO4 3--P concentrations ranged from 0.43 to 16.00μM, 1.50 to 132.38μM, and 1.30 to 6.29μM, respectively; the values at some stations were much higher than observed in Tongyeong coast seawater. Using seawater from station B, C. polykrikoides grew consistently, with a high growth rate, similar to the red tide in nature. This seawater appeared to contain materials needed by C. polykrikoides to grow. Therefore, C. polykrikoides red tides seem to occur wherever the ground sea water contains materials that are needed for its growth.

Harmful dinoflagellate Cochlodinium polykrikoides blooms have been frequently occurred in coastal areas of the East Sea since 1995. We compared the oceanic conditions in years 1995, 2001 and 2003 when the C. polykrikoides bloom was strong, and in years 1998 and 2004 when the C. polykrikoides bloom was not appeared. We studied temporal and spatial variation of upwelling and geostrophic currents on the western channel of Korean Strait, an entrance of the East Sea. The period and occurrence area of C. polykrikoides bloom was depended on variation of upwelling in summer. In the distributions of geostrophic current, southward current was dominant near the coast in August, 1998 and 2000. Whereas northward current was dominant near and off the coast in August, 1995 and 2003 which the C. polykrikoides bloom was strong. When compared dominant phytoplankton of the coastal areas in each year, Kuroshio indicator species Proboscia alata and Chaetoceros affine were dominant, respectively, in 2001 and 2003 at every stations. However, the dominant species was variable at each coastal area in 1998 and 2000. In 2003, the abundance of Sagitta elegans which is known as the cold water indicator was low, but the abundance of S. enflata, warm water indicator, was very high in Gangneung compared to Sokcho. It seemed that the distribution of S. elegans is restricted by strong warm water current. In conclusion, it was estimated that the distribution of C. polykrikoides bloom in the coastal area of the East Sea was closely related with the strength of East Korea Warm Current and upwelling.

Eco-physiological research and the control of Cochlodinium polykrikoides was carried out to elucidate eco-physiological characteristics of red tide organism through culture experiment depending on the condition of photon irradiance. Oxygen consumption of C. polykrikoides was high with a value of 1.12mg/L/hr in the dark compared with that of 0.13mg/L/hr at 100μEm-2s-1. DO values in a circular chamber with the lapse of time in seawater containing C. polykrikoides were declined in the dark period. DO values of seawater containing C. polykrikoides in the dark were declined from 7.01mg/L to 2.65mg/L in 30cm depth and from 7.01mg/L to 6.63mg/L in 5cm depth depending on the depth of circular culture vessel. Olive flounder, Paralichthys olivaceus and file fish, Stephanolepis cirrhifer exposed to Cochlodinium showed the separation of the lamella epithelium from gill filament, which disrupted the respiratory process at the gill level.

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℃, 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 × 104 cells/㎖) cultures, 100% of C. polykrikoides cells was destroyed within 15 hours. The released algicides were heat-tolerant to 100℃ and stable in pH 6.0～10.0. These results suggest that Brachybacterium sp. SY-97 is potentially useful for controlling outbreaks of C. polykrikoides.

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 381T. Therefore, this bacterium was designated Micrococcus luteus SY-13. The optimal culture conditions of the algicidal bacterium was 25℃, 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 × 104 cells/㎖) 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℃ 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∼180℃ 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.