• 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.

• 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.

• Proceedings of the Korean Environmental Sciences Society Conference
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• 2008.11a
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• pp.293-296
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• 2008

적조발생 해역인 울진 연안의 해수에서 분리한 100여종의 해양미생물 중 8 균주가 C. polykrikoides에 대해 살조활성을 나타내었으며, 이 중 살조활성이 가장 우수한 Brachybacterium sp. SY-97을 선별하였다. Brachybacterium sp. SY-97의 최적 배양조건은 30$^{\circ}C$, pH 7.0, 2.0% NaCl 농도였다. Brachybacterium sp. SY-97은 C. polykrikoides를 살조시킨 후 생성되는 EOM을 이용하여 성장하며, 유도기를 거쳐 대수증식기에 살조물질을 활발히 생산하는 것으로 판단된다. 0.2 $\mu$m의 Cell Culture Insert를 삽입한 2조 배양계를 이용하여 Brachybacterium sp. SY-97의 살조 유형을 조사한 결과, Brachybacterium sp. SY-97은 0.2 $\mu$m filter에 의해 C. polykrikoides와 격리된 상태에서도 C. polykrikoides를 살조시켜 직접 공격형이 아니라 세포외로 물질을 분비하여 살조시키는 살조인자 분비형으로 밝혀졌다. Brachybacterium sp. SY-97의 배양여과액의 첨가 농도별 살조활성을 측정한 결과, 15%의 경우 C. polykrikoides의 개체수는 급격히 감소하여 12시간 후에 100% 살조되었고, 10%의 경우 15시간 후에 모두 살조되었고, 5%의 경우 18시간 후에 control에 비해 90% 이상 살조 되었다.

• Korean Journal of Fisheries and Aquatic Sciences
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• v.32 no.4
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• pp.452-457
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• 1999

Algicidal bacteria were isolated from the mud of south coastal sea of Korean peninsula and screened to evaluate algicidal activity on Cochlodinium polykrikoides. The optimum condition for the development of highest algicidal activity was determined, The optimum sodium chloride concentration for algicidal activity of isolated algicidal bacteria was $3\%$. The optimum temperature, pH and culture time for the highest algicidal activity were $30^{\circ}C$, pH 7 and above 24 hr, respectively. The algicidal activity were significantly increased at the stationary phase of the cell growth and continuously increased up to maximum at the death phase, probably due to the release of algicidal substance by cell Iysis. The effect of zinc ion addition on algicidal activity, was observed and indicated that the substance requires zinc for its activity. The candidate for the algicidal substance may be $\alpha$-mannosidase.

• Ecology and Resilient Infrastructure
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• v.3 no.1
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• pp.22-34
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• 2016

We developed a biologically-derived substance naphthoquinone (NQ) derivate for the eco-safe mitigation of harmful cyanobacteria blooms such as Microcystis and Dolichospermum. NQ was reacted with various substituents ($R_n$) to produce different NQ derivatives. We tested a total of 92 algicidal compounds based on the algicidal activity of Microcystis and Dolichospermum. 22 compounds of NQ were selected as candidates (algicidal activity >80% at $1{\mu}M$). Among them, NQ 40 compound showed the highest algicidal activity of 99.6% and 100% at the optimal concentration of $1{\mu}M$ on Microcystis and Dolichospermum, respectively. No algicidal effects of NQ 40 ($1{\mu}M$) were observed against non-target algae such as Stephanodiscus, Cyclotella and Peridinium. According to the results of acute eco-toxicity assessment, the $EC_{50}$ values of NQ 40 compound for Selenastrum capricornutum and Daphnia magna were 3.2 and $14.5{\mu}M$, respectively, and the $LC_{50}$ for Danio rerio was $15.7{\mu}M$. In addition, for D. magna chronic eco-toxicity assessment, no toxicity toward survival, growth and reproduction was observed. Therefore, we suggested the NQ 40 ($1{\mu}M$) compound as an alternative eco-safe algicidal substance to effectively mitigate harmful cyanobacteria blooms.

• Korean Journal of Environmental Biology
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• v.38 no.1
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• pp.114-126
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• 2020

Bloom-forming toxic cyanobacteria Microcystis spp. are common in the summer season in temperate freshwater ecosystems. Often, it leads to the degradation of water quality and affects the quality of drinking water. In a previous study, NQ (naphthoquinone) compounds were shown to be effective, selective, and ecologically safe algicides for Microcystis spp. blooms. To analyze the superiority of developed NQ derivatives, we conducted a microcosm experiment using clay, which is frequently used in South Korea. Similar to previous studies, the NQ 40 and NQ 2-0 compounds showed high algicidal activities of 99.9% and 99.6%, respectively, on Microcystis spp. at low concentrations (≥1 μM) and enhanced phytoplankton species diversity. However, when treated with clay, a temporary algicidal effect was seen at the beginning of the experiment that gradually increased at the end. In addition, treatment with the NQ compounds did not affect either the abiotic or biological factors, and similar trends were observed with the control. These results showed that the NQ 2-0 compound was more effective, with no ecosystem disturbance, and more economical than the currently used clay. These results suggest that NQ 2-0 compound could be a selective, economically and ecologically safe algicide to mitigate harmful cyanobacterial blooms in the field.

• Korean Journal of Environmental Agriculture
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• v.31 no.2
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• pp.175-184
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• 2012

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. METHODS AND RESULTS: Among of four algicidal bacteria isolated in this study, NH-12 isolate was the strongest algicidal activity against A. catenella. NH-12 isolate was identified on the basis of biochemical characteristics and analysis of 16S rRNA gene sequences. The isolate showed 97.67% homology with Pseudoalteromonas prydzensis ACAM $620^T$ (U85855), and was designated Pseudoalteromonas sp. NH-12. The optimal culture conditions of this isolate were $25^{\circ}C$, initial pH 8.0, and 3.0% (w/v) NaCl concentration. The algicidal activity of NH-12 was significantly increased to maximum value in the late of logarithmic phase of bacterial culture. As a result of 'cell culture insert' experiment, NH-12 is assumed to produce secondary metabolites, as an indirect attacker. When 10% culture filtrate of NH-12 was applied to A. catenella, over 99% of algal cells were destroyed within 24 h. In addition, the killing effects were increased in dose and time dependent manners. CONCLUSION(S): Taken together, our results suggest that Pseudoalteromonas sp. NH-12 could be a candidate for controlling of toxic algal blooms.

• Ecology and Resilient Infrastructure
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• v.7 no.1
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• pp.72-81
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• 2020

Blooms of the small centric diatom Stephanodiscus is quite occasional in winter season in temperate freshwater ecosystems. Often, it leads to degradation of water quality and affects quality of supplied drinking water. In previous studies, naphthoquinone (NQ) compounds have been shown to be effective and selective for controlling winter bloom species Stephanodiscus hantzschii. We conducted a 5 ton scale mesocosm experiment to investigate the effects of NQ on native Stephanodiscus sp. collected from Nakdonggang River in water. After treatment with NQ 4-6 compound (0.2 μM), the cell density of Stephanodiscus sp. was rapidly reduced from 5 × 10³ cells mL^-1 to 0.2 × 10³ cells mL^-1 for 10 days. Additionally, NQ 4-6 compound did not affect physicochemical factors (water temperature, dissolved oxygen, pH, conductivity, nutrients) and biological factors (bacteria, heterotrophic nanoflagellates, zooplankton). Therefore, these findings suggest that the NQ 4-6 compound has potential as an alternative algicidal substances to effectively mitigate natural Stephanodiscus sp. blooms, and the application of NQ 4-6 compound will restore the healthy aquatic ecosystems.

• Korean Journal of Environmental Biology
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• v.36 no.3
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• pp.359-369
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• 2018

Harmful algal blooms (HABs) are a serious problem for public health and fisheries industries, thus there exists a need to investigate the possible ways for effective control of HABs. In the present study, we investigated the algicidal effects of a newly developed GreenTD against the HABs (Chattonella marina, Heterosigma akashiwo, Cochlodinium polykriokides, and Heterocapsa circularisquama) and non-HABs (Chaetoceros simplex, Skeletonema sp. and Tetraselmis sp.), which is focused on the different population density and concentration gradients of algicidal substances. The time series viability of target alga was assessed based on the activity of Chl. a photosynthetic efficiency in terms of $F_v/F_m$, and in vivo fluorescence (FSU). Effective control of Raphidophyta, C. marina and H. akashiwo was achieved at a GreenTD concentration of $0.5{\mu}gL^{-1}$ and $0.2{\mu}gL^{-1}$, respectively, and regrowth of both the species was not observed even after 14 days. The inhibitory ratio of the dinoflagellate, C. polykriokides was more than 80% at $0.2{\mu}gL^{-1}$ of GreenTD. H. circularisquama was constantly affected in the presence of $0.2{\mu}gL^{-1}$ of GreenTD in the high- and low-population density experimental groups. On the other hand, diatoms, C. simplex, and Skeletonema sp. were not significantly affected even in the presence of $0.2{\mu}gL^{-1}$ of GreenTD and exhibited re-growth activity with the passage of incubation time. In particular, green alga Tetraselmis sp. remained unaffected even in the presence of the highest concentration of GreenTD ($1.0{\mu}gL^{-1}$), implying that non-HABs were not greatly influenced by the algicidal substances. As a result, the algicidal activity of GreenTD on the harmful and nonharmful algae was as follows: raphidophyte>dinoflagellates>diatoms>green alga. Consequently, our results indicate that inoculation of GreenTD substances into natural blooms at a threshold concentration ($0.2{\mu}gL^{-1}$) can maximize the algicidal activity against HABs species. If we consider the dilution and diffusion rate in the field application, it is hypothesized that GreenTD will demonstrate economic efficiency, thus leading to effective control against the target HABs in the closed bay.

• Korean Journal of Ecology and Environment
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• v.37 no.2 s.107
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• pp.241-247
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• 2004

Isolation and identification of alga-lytic bacteria were carried out. Fifteen isolates of alga-lytic bacteria were screened by the double layer method using A. cylindrica NIES-19 as a sole nutrient and four isolates among them were compared with their alga-lytic activity. The isolate AK-05 exhibiting the highest alga-lytic activity was identified as Acidovorax temperans base on its 16S rDNA sequence. The culture supernatant of the isolate AK-05 was reliable for the alga-lytic. Alga-lytic activity assays of culture supernatant revealed that the major substances for alga-lytic activity were non-proteins and heat stable. The highest alga-Iytic activity was practical under alkaline conditions and at 25${\sim}$$30^{\circ}C$. It is indicating an advantage for the application of water blooms by cyanobacteria in eutrophic lakes where the pH is generally in alkaline region.