• 한국환경과학회지
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• 제17권4호
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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.

• 한국환경농학회지
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• 제34권2호
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• pp.139-148
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• 2015

BACKGROUND: The aim of this study was to isolate and identify algicidal bacterium that tends to kill the toxic dinoflagellate Alexandrium catenella, and to determine the algicidal activity and algicidal range of algicide. METHODS AND RESULTS: Among of algicidal bacteria isolated in this study, NH-3 isolate was the strongest algicidal activity against A. catenella. NH-3 isolate was identified on the basis of biochemical characteristics and analysis of 16S rRNA gene sequences. The NH-3 isolate showed over 99% homology with Arthrobacter oxydans, and was designated as Arthrobacter sp. NH-3. The optimal culture conditions were $25^{\circ}C$, initial pH 7.0, and 2.0% (w/v) NaCl concentration. The algicidal activity of Arthrobacter sp. NH-3 was significantly increased to maximum value in the late of logarithmic phase. Arthrobacter sp. NH-3 showed algicidal activity through indirect attack, which excreted active substance into the culture filtrate. When 10% culture filtrate of NH-3 was applied to A. catenella, 100% of algal cells were destroyed within 30 h. In addition, the algicidal activities were increased in dose and time dependent manners. The pure algicide was isolated from the ethyl acetate extract of the culture filtrate of NH-3 by using silica gel column chromatography and high performance liquid chromatography (HPLC). We investigated the algicidal activity of this algicide on the growth of harmful algal bloom (HAB) species, including A. catenella. As a result, it showed algicidal activity against several HAB species at a concentration of $100{\mu}g/mL$ and had a relatively wide host range. CONCLUSION: Taken together, our results suggest that Arthrobacter sp. NH-3 and its algicide could be a candidate for controlling of toxic and harmful algal blooms.

• 한국수산과학회지
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• 제32권4호
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• pp.452-457
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• 1999

적조 현상이 나타난 남해안 연안의 뻘로부터 Cochlodinium polykrikoides에 대한 살조능을 가지는 살조 세균 KY1을 분리하였다. 분리된 살조 세균 KY1이 살조능을 나타내는 최적 조건은 $30^{\circ}C$, 염농도 $3\%$, pH 5 이상의 중성으로 확인되었으며, 약 8 시간의 배양에서 대수 증식의 최대 성장률을 보였으며, 이후 사멸기 까지 배양 시간 24 시간 경과 후부터 높은 살조능을 나타내었다. 살조능은 대수증식기에서 정지기로 넘어갈 때, 큰 폭으로 증가하였다. 그리고, 사멸기에서는 최대 $100\%$에 달하는 살조능을 나타내었다. 이는 살조 세균이 분해되면서 균체내에 존재하는 살조물질을 분비하는 것으로 간주된다. 또한 배양 상등액의 양을 증가시켰을 때, 1 시간 이내에 $40{\mu}{\ell}$/ml의 농도에서 $100\%$의 살조능을 볼 수 있었다. $Zn^{2+}$의 첨가로 살조능의 현저한 증가를 확인 할 수 있었으며, 이로부터 살조물질은 C. polykrikoides의 세포벽 성분인 mannan을 분해하는 $\alpha$-mannosidase일 가능성을 보였다.

• 한국수산과학회지
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• 제40권3호
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• pp.122-127
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• 2007

Photosensitized extracts of 28 seaweed species were tested in vitro to examine the variation in their algicidal activities against the toxic microalga Cochlodinium polykrikoides. When both seaweed extracts and microalgae were exposed to UV and visible light, methanol extracts of Porphyra yezoensis showed 5.7- and 4.4-fold increasesin light-dependent algicidal activity, respectively. When only seaweed extracts were exposed to both UV (365 nm) and visible light (white lamp) at the same time, methanol extracts of Enteromorpha linza and Carpopeltis affinis showed 3.3- and 3.4-fold increases in algicidal activity, respectively. When UV-photosensitized extracts were left in the dark, the algicidal activity of Ecklonia cava increased 13-fold after 5 h. When visible light-photosensitized extracts were left in the dark, the algicidal activity of Monostroma nitidum increased by 3.3-fold in 1 h.

• Fisheries and Aquatic Sciences
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• 제5권3호
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• pp.150-155
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• 2002

Marine bacteria producing algicidal substance against Cochlodinium polykrikoides was screened and isolated from seawater. Metabolite of marine bacteria Micrococcus sp. LG-5 showed strong algicidal activity against C. polykrikoides. C. polykrikoides was inhibited above $90\%$ at $5\%$ solution of the metabolite within 24 hrs. Algicidal substance from the metabolite was extracted with ethyl acetate, and then purified by PTLC and reversed-phase HPLC. Algicidal activity of purified compound against C. polykrikoides was above $90\%$ at 3.7, 11.0 and 33.0${\mu}g/mL$ concentration after 12, 9 and 3 hrs, respectively. Ninety percent inhibition of other red tides, Gymnodinium sanguineum and Gyrodinium impudicum was observed when treated with 3.7${\mu}g/mL$ of purified compound within a period of 12 hrs. The microscopic view of red tides treated with purified compound showed the deformations such as cell node cuts and swelling of cells.

• 한국수산과학회지
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• 제32권2호
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• pp.160-164
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• 1999

해양 세균을 이용하여 규조류, Chaetoceros spp. 적조의 방제기술 개발을 위한 목적의 일환으로 우리나라 연안에서 분리된 Alteromonas sp. SR-14가 여러 가지 조건하에서 Chaetoceros calcitrans의 증식에 미치는 영향을 조사하였으며 그 결과를 요약하면 다음과 같다. Alteromonas sp. SR-14를 C. calcitrans와 혼합배양하였을 때 조류 증식 저해활성은 $20^{\circ}C$에서는 강하였으나 $25^{\circ}C$ 이상에서는 활성이 없었다. 그리고 C. calcitrans의 초기농도가 약 $10^4$ cells/ml이었을 때 Alteromonas sp. SR-14를 10 cells/ml 이하만 접종하여도 증식을 저해하였다. Alteromonas sp. SR-14는 대수기의 C. calcitrans에 대해서 저해 활성이 강하였고, 무기 영양염의 첨가나 1,300$\~$4,600 lux에서 조도변화 및 200 rpm으로의 진탕 등의 배양조건은 세균의 조류증식 저해활성에 영향을 미치지 않았다.

• 한국수산과학회지
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• 제34권5호
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• pp.430-434
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• 2001

미세조류 증식 저해균 Shewanella sp. SR-14와 연안해역에서 우점종으로 분포하는 세균인 Vibrio alginolyticus가 규조, Chaetoceros calcitrans의 증식에 미치는 영향 그리고 V. ajginolyticus가 Shewanella sp. SR-14의 C. calcitrans 증식저해 활성에 미치는 영향을 혼합배양으로 측정하였다. Shewsnella sp. SR-14와 해수에서 분리한 V. alginolyticus는 미세조류 배양용 무기배지에서 단독 또는 혼합 배양하여도 잘 증식하였으며, C. calcitrans와 혼합배양시 Shewanella sp. SR-14는 조류증식을 저해한 반면 V alginolyticus는 이 조류의 증식을 촉진하였다. 조류가 첨가된 배지에 Shewanella sp. SR-14의 최초 접종균수가 V. alginolyticus와 거의 같거나 1 log 정도 적게 접종되었을 때에는 C. calcitrans에 대한 Shewanella sp. SR-14의 증식 저해활성이 유지되었으나 V. alginolricus의 최초균수가 Shewanella sp. SR-14보다 3 log 이상 많았을 때에는 조류증식 저해활성이 나타나지 않았다.

• 한국수산과학회지
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• 제33권4호
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• pp.331-338
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• 2000

최근 남해안 적조의 주요 원인생물인 C. polykrikoides를 살조 시키는 해양세균을 마산만의 적조발생 해역에서 분리, 동정하고 살조범위를 조사하였다. 분리 균주 중 살조활성이 우수한 Micrococcus sp. LG-5의 성장 특성과 살조효과에 대해 조사하여 해양 미생물을 이용한 자연생태 조화형, 환경 친화적 적조방제 기술 개발의 기초 자료를 제공하고자 연구한 결과를 요약하면 다음과 같다. 살조세균 Micrococcus sp. LG-5의 최적 성장조건은 $25^{\circ}C, NaCl 3.0{\%}, pH 7.0$이었으며, C. polykrikoides ($1.2{\times}10^4 cells/ml$)에 대한 Micrococcus sp. LG-5의 성장단계에 따른 배양여과액의 살조활성은 대수증식기 후기 및 정지기에 강하였다 대수증식기에는 C. polyklikoides를 $100{\%}$, 정지기에는 $82{\~}92{\%}$, 대수증식기 중기에는 $40{\~}80{\%}$, 유도기에는 $5{\%}$ 이하의 살조활성을 보여 Micrococcus sp. LG-5는 유도기를 거쳐 대수증식기에 살조물질을 활발히 생산하는 것으로 밝혀졌다. 또한 $0.4{\mu}m$의 cell culture insert를 삽입한 2조배양계를 이용하여 Micrococcus sp. LG-5의 살조유형을 조사한 결과, Micrococcus sp. LG-5는 $0.4{\mu}m filter$에 의해 C. polykirkoides와 격리된 상태에서도 C. polykirkoides를 살조시켜, 세포외로 물질을 분비하여 살조시키는 살조물질 분비형으로 밝혀졌다. 또한 Micrococcus sp. LG-5의 배양여과액의 최종농도가 $5{\%}, 10{\%}$가 되도록 첨가한 경우, C. polykrikoides의 개체수는 급격히 감소하여 3시간 경과 후에 centrol에 비해 각각 $30{\%}$와 $85{\%}$정도가 살조되었으며, $1{\%}$의 경우, 30시간 후에 $90{\%}$ 이상 살조되었다. 즉, Micrococcus sp. LG-5의 배양여과액의 양에 비례하여 살조효과가 증가하는 경향을 보였으며, 배양여과액 중에 C. polykrikoides를 사멸시키는 살조물질이 존재함을 알 수가 있었다.

• 한국환경과학회지
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• 제18권4호
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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.

• 생태와환경
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• 제49권3호
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• pp.176-186
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• 2016

For several decades, lactic acid bacterium (Lactobacillus graminis: LAB) has been generally recognized as safe. To develop the pan-environmental bio-control agent, algicidal activity of the live LAB cell and its culture filtrate (CF) was examined against Microcystis aeruginosa. LAB cells perfectly lysed M. aeruginosa within 3 days, while the CF had a less effect than the live cells, approximately 78% inhibition of algal growth during a same culture period. The concentration of microcystin in alone culture of M. aeruginosa was $7.1{\mu}gL^{-1}$, but gradually increased and leach $158.5{\mu}gL^{-1}$ on 10 days. However, LAB cells clearly decreased the microcystin by $10.3{\mu}gL^{-1}$ in the same period, approximately 93.5%. CF of LAB showed a strong algicidal activity over 75% between pH 2-7, 91.3% by the treatment of proteinase K, 87.8% by below 3 kDa in particle size, and 75.3% by heat treatment, respectively. Of five solvents, fractions of CF passed through solvents diethyl ether and ethyl acetate showed an obvious algicidal activity in the algal-lawn test. Among 5 fractions purified by silica-gel TLC plate, two spots showed a most strong removal activity on M. aeruginosa. Another analysis of GC indicate that CF contained six representative fatty acids. Even though most of these substance have been known as an anti-algal substance against M. aeruginosa, oleic acid is the most effective. These results suggested that the culture filtrate or specific substances, like a fatty acids, in comparison with live L. graminis can be a successful and eco-friendly agent to control Microcystis bloom.