• Journal of Korean Society on Water Environment
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• v.34 no.2
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• pp.210-215
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• 2018

It is difficult to count colonial cyanobacteria Microcystis cells since the thickness of colonies is constrained by amorphous mucilage, making it impossible to estimate the number of cells. Disaggregation of Microcystis colonies into single cell is needed to improve the accuracy and precision of cell density estimation of naturally collected samples. Uultrasonic treatment method is commonly used owing to the simplicity and immediacy of the procedure. However, amplitude, frequency, and duration of ultrasonic treatment also cause cell loss during the experiment. Optimal ultrasonic treatment has not been standardized yet. Therefore, the objective of this study was to investigate optimal ultrasonic treatment by analyzing cell density and colony numbers. We collected colonial Microcystis from Changnyeong-Haman weir area in Nakdong River during harmful algal boom period from September to October in 2017. Ultrasonic treatment method was applied to disrupt colonies into single cells to enumerate cell density. Among treatment conditions, results from continuously treated for 100 seconds were found to be the optimum to reduce colonies to a suspension of single cell without cell losses under high and low density of Microcystis cells. Lugol iodine fixed cells followed by sonication showed less negative impact of cell damage within the optimal treatment time (100 seconds). Furthermore, disaggregated cells treated by sonication enables microscopic observation more easily since gas vacuoles were collapsed to facilitate sedimentation of cells under the counting chamber for quantitative enumeration of buoyant Microcystis cells.

• Journal of Microbiology
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• v.44 no.5
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• pp.562-565
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• 2006

The cell counting of colonial Microcystis spp. is a rather difficult and error-prone proposition, as this genus forms irregularly-shaped and irregularly-sized colonies, which are packed with cells. Thus, in order to facilitate a cell count, four methods of dividing the colonies into single cells were compared, including vortexing, sonication, $TiO_2$ treatment, and boiling. As a result, the boiling method was determined to generate the greatest number of single cells from a colony, and all colonies were found to have divided completely after only 6 min of treatment. Furthermore, no significant cell destruction, which might alter the actual cell density, was detected in conjunction with the boiling method (P=0.158). In order to compute the cell number more simply, the relationship between the colony size and the cell number was determined, via the boiling method. The colony volume, rather than the area or diameter was correlated more closely with the cell number ($r^2=0.727$), thereby suggesting that the cell numbers of colonial Microcystis sp. can also be estimated effectively from their volumes.

• Proceedings of the Zoological Society Korea Conference
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• 1998.10b
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• pp.189.2-189
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• 1998

No Abstract, See Full Text

• Journal of Microbiology
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• v.40 no.4
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• pp.335-339
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• 2002

Partial sequences of 16S rRNA gene of five chroococcalian blue-green algal strains, Aphanothece nidulans KCTC AG10041, Aphanothece naegelii KCTC AG10042, Microcystis aeruginosa KCTC AG10159, Microcystis ichthyoblabe KCTC AG10160, and Microcystis viridis KCTC AG10198, which were isolated from water from the Geumgang River, were determined and were inferred their phylogenetic and taxonomic positions among taxa of order Chroococcales. Most taxa of Chroococcales whose partial 16S rRNA gene sequences were aligned in this study, are clustered with other related taxa. Aphanothece nidulans KCTC AG10041 and Aphanothece naegelii KCTC AG10042 made a cluster with other European species of these genera, which supported 100% of the bootstrap trees with a very high sequence similarity (97.4-99.4%) in this study. Three strains, Microcystis aeruginosa KCTC AG10159, M. ichthyoblabe KCTC AG10160, and M. viridis KCTC AG10198, formed a cluster with other Microcystis spp. supported 100 % of the bootstrap trees with a similarity of 97.0-99.9% except for two strains. However, this phylogentic tree made no resolution among the species of Microcystis spp. The topology of the tree reconfirmed the taxonomic status of three species of Microcystis, identified in this study based on the morphology, as three colonial types of Microcystis aeruginosa com. nov. Otsuka et al. (1999c). The genera of chroococcalian cyanophytes are heterogeneously clustered in these sequence analyses. We suggest that more molecular studies on the genera of Chroococcales with reference strains, widely collected from restricted geographic or environmental ranges, get accurate taxonomic or phylogenetic determinations.

• Korean Journal of Ecology and Environment
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• v.42 no.1
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• pp.75-84
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• 2009

The effect of various SNPs (silver nanoparticles) on the growth of Microcystis aeruginosa was investigated in laboratory and field experiment. Four SNPs, namely JS47N, JS47N-K2, JS47N/3-1 and JS47N/3-2 were used to this study. The Ag size, concentration and color of these solutions were about $20{\sim}40nm$, $200mg\;L^-1$ and brown, respectively. At 0.01 and $0.1mg\;L^-1$, SNPs inhibited the growth of unicellular M. aeruginosa by 99.4% and 99.9%, respectively. However, SNPs of $1mg\;L^-1$ inhibited the growth of colonial M. aeruginosa by 98.5%, whereas the other three concentrations (0.001, 0.01 and $0.1mg\;L^-1$) had little inhibitory effect. In experimental enclosures from eutrophic lake, cyanobacteria including M. aeruginosa were found to be more sensitive to the SNPs than green algae and diatoms. In conclusion, our study indicates that SNPs has a selective cyanocidal potential when used to M. aeruginosa. We believe that future studies need to test on various other organisms, and determine minimum concentration for field application.

• Korean Journal of Environmental Biology
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• v.31 no.2
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• pp.113-120
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• 2013

This study aimed to investigate the effects of ultrasonic wave on colonial changes of Microcystis sp. causing water-bloom. The effects of frequency change and exposure time of ultrasonic wave on Microcystis sp. were also perceived by measuring the concentration of chlorophyll-a. The concentrations of chlorophyll-a showed a decreasing tendency in all experimental groups exposed to ultrasonic waves, especially the highest decrease at 70 kHz. Before exposed to ultrasonic wave, single cells of Microcystis sp. were observed in a form of colonies which were densely accumulated onto their mucilage layer. However, after one hour exposure to 70 kHz ultrasonic wave, the bonds of colonies were weakened and single cells were separated from the colonies. Moreover, the single cells of the colony were destroyed with the 5 hour exposure time at 70 kHz. As a result, we proposed that the frequency strength and the exposure time of ultrasonic wave might have effects in decrease of biomass of blue-green algae by reducing the concentration of chlorophyll-a, thereby changing and destroying forms of Microcystis sp. colonies.

• Korean Journal of Ecology and Environment
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• v.38 no.3 s.113
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• pp.420-428
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• 2005

Effects of three biological control agents such as Xanthobacter autotrophycus, Tanichthys albonubes and Oryzias latipes on the morphology and growth of cyanobacterium Microcystis aeruginosa were studied. The experiments were consisted of six treatments of living organism (LO) and culture filtered water of three organisms (CFW). Three LOs effectively decreased the density of M. aeruginosa, and then cyanobacteria hardy showed in the microscopic field after 5 days of cultivation. All LO and CFW agents induced the colonial formation of cyanobacterium M. aeruginosa, although there were little differences in colony formation according to the kinds, density and type of treatment. In particular, the higher density treatment of fish CFW induced effectively the colony formation of cyanobacteria, compared to the bacterial LO and CFW. Thus, the application of bio agents to control the cyanobacterial bloom is needed to the further study to diminish the adverse effects such as the enhancement of colony formation towards on the new bloom against the aquatic ecosystem.

• Korean Journal of Ecology and Environment
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• v.41 no.spc
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• pp.77-85
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• 2008

Grazing rates (GR) and pseudofaeces production (PFP) of native snail, Chinese mystery snail (Cipangopaludina chinensis malleata Reeve) on natural colonial morphs of Microcystis aeruginosa was measured. C. chinensis was collected from the upstream of the Geum River (Boryeong, Korea), where they co-habituated with Unio douglasiae and Lanceolaria acrorhyncha. The experiments were performed to evaluate the GR and PFP at different conditions such as; incubation time (1, 3, 5, 7, 9 and 11 hr), body size (3 to 6.1 cm, n=28), snail density (0.5, 1, 1.5 and 2.0 ind. $L^{-1}$) and prey concentration (168.3, 336.7, 505.0 and $673.0{\mu}g\;Chl-{\alpha}L^{-1}$). All experiments were triplicated, and conducted in transparent acrylic vessel (3L in volume). Regarding feeding time, a highest GR (0.538L $gAFDW^{-1}h^{-1}$) and PFP $(7.18mgAFDW^{-1})$ appeared at 1hr and 7hr after snail stocking, respectively. Interestingly, the snail, smaller than 4.5cm in body size, showed a wide range of GR ($-4.173{\sim}1.087L\;gAFDW^{-1}h^{-1}$) for the initial period (1 and 4hrs of stocking), compared to those greater than 4.5cm, which showed a stable FR, higher than 0.5L $gAFDW^{-1}h^{-1}$. Upon density effect, the density of 1.5 ind. $L^{-1}$ induced the most effective inhibition on Microcystis biomass with highest PFP. On the prey concentration, highest GR (0.897L $gAFDW^{-1}h^{-1}$) and PFP (3.67 mg $gAFDW^{-1}h^{-1}$) were induced at the level of $168.3{\mu}g\;Chl-{\alpha}L^{-1}$ and $673{\mu}g\;Chl-{\alpha}L^{-1}$, respectively. GR and PFP of this freshwater snail on the cyanobacterial bloom (M. aeruginosa) varied with the feeding conditions, and they were comparatively high for a short period of time less than 7hrs regardless of the stocking condition. Our results suggest that this freshwater snail has a potential to control cyanobacterial bloom when provided with suitable condition.

• Korean Journal of Ecology and Environment
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• v.43 no.2
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• pp.221-231
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• 2010

A simple extraction method was applied to control four selected cyanobacteria, solitary (SMA) and colonial Microcystis aeruginosa (CMA), and green algae, Scenedesmus quadricauda and Chlorella vulgaris using a domestic plant and mineral. Three kinds of concentrations (1, 5, and 10 mg $L^{-1}$) of three fresh plants Camellia sinensis, Quercus acutissima, and Castanea crenata, three minerals loess, quartz porphyry, and natural zeolite, and plant-mineral composite, totally seven materials were prepared with the simple extraction processes: drying and grinding of material, water-extraction by high temperature-sonication and filtering. Cyanobacteria SMA and CMA (over 60% of control) were effectively inhibited with the low concentration (1 mg $L^{-1}$) of plants Q. acutissima and C. crenata and natural zeolite, while green alga S. quadricauda (below 50% of control) also retarded in growth. Low concentrations (1 mg $L^{-1}$) of C. sinensis effectively increased the growth of C. vulgaris, while loess also induced the algal growth of S. quadricauda. Therefore, our results indicate that crude extract of domestic plants, Q. acutissima and C. crenata can be also useful to control the cyanobacterial bloom in eutrophic lake, whereas C. sinensis and loess may be a good growth factor or useful media for the algal mass culture.

• Korean Journal of Environmental Biology
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• v.20 no.1
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• pp.91-99
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• 2002

With physico-chemical environmental factors such as temperature, DO, pH, conductivity and nutrients, size fractionated of phytoplankton standing crops and chlorophyll a concentrations $(>8\mu{m},\;3-8\mu{m},\;<3\mu{m})$ and their relationships were investigated at each station of Kyungan Stream and Togyo Reservoir from April to November in 1997. The two sampling sites showed different nutrient status: Kyungan Stream was eutrophic, while Togyo Reservoir was mesotrophic. Large sizes of phytoplankton and chl. a were higher in Kyungan Stream, opposite to those of Togyo Reservoir; Standing crops of phytoplankton $(>8\mu{m)$ and chl. a $(3-8\mu{m)}$ were high in Kyung-an Stream, while phytoplankton $(3-8\mu{m)}$and chi. a $(<3\mu{m)$ were abundant in Togyo Reservoir. These results imply that phytoplankton community in the highly eutrophicated water mainly comprised the large filamentous and/or colonial algae, such as Microcystis spp. and Aphanizomenon flos-aquae, which easily enriched by nutrients loading.