• Environmental Engineering Research
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• v.24 no.3
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• pp.397-403
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• 2019

Microcystis sp. is one of the most common harmful cyanobacteria that release toxic substances. Counting algal cells is often used for effective control of harmful algal blooms. However, Microcystis sp. is commonly observed as a colony, so counting individual cells is challenging, as it requires significant time and labor. It is urgent to develop an accurate, simple, and rapid method for counting algal cells for regulatory purposes, estimating the status of blooms, and practicing proper management of water resources. The flow cytometer and microscope (FlowCAM), which is a dynamic imaging particle analyzer, can provide a promising alternative for rapid and simple cell counting. However, there is no accurate method for counting individual cells within a Microcystis colony. Furthermore, cell counting based on two-dimensional images may yield inaccurate results and underestimate the number of algal cells in a colony. In this study, a three-dimensional cell counting approach using a novel model algorithm was developed for counting individual cells in a Microcystis colony using a FlowCAM. The developed model algorithm showed satisfactory performance for Microcystis sp. cell counting in water samples collected from two rivers, and can be used for algal management in fresh water systems.

• ALGAE
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• v.19 no.2
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• pp.149-151
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• 2004

Freshwater algae make up a very important portion of the autotrophic component of the aquatic food web. Therefore, the study of freshwater algal structure and biomass is central to aquatic ecosystem studies. Due to variations in cell shape and size for each species (or taxon) and survey site, cell abundance (or cell numbers per chosen volume) often leads to misrepresentation of the true importance of some species because of the great differences in size of various algae. Thus, it is necessary to investigate the freshwater algal species of a site in order to calculate the cell volume. Although direct cell counting, species volume measurement, as well as biomass calculation are time-consuming and requiring specialists in taxonomy.

• ALGAE
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• v.37 no.4
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• pp.281-291
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• 2022

Quantifying the abundance of Chattonella species is necessary to effectively manage the threats from ichthyotoxic raphidophytes, which can cause large-scale mortality of aquacultured fish in temperate waters. The identification and cell counting of Chattonella species have been conducted primarily on living cells without fixation by light microscopy because routine fixatives do not retain their morphological features. Species belonging to the Chattonella marina complex, including C. marina and C. marina var. ovata, had high genetic similarities and the lack of clear morphological delimitations between the species. To estimate the abundance of C. marina complex in marine plankton samples, we developed a protocol based on the droplet digital polymerase chain reaction (ddPCR) assay, with C. marina complex-specific primers targeting the internal transcribed spacer (ITS) region of the rDNA. Cell abundance of the C. marina complex can be determined using the ITS copy number per cell, ranging from 25 ± 1 for C. marina to 112 ± 7 for C. marina var. ovata. There were no significant differences in ITS copies estimated by the ddPCR assay between environmental DNA samples from various localities spiked with the same number of cells of culture strains. This approach can be employed to improve the monitoring efficiency of various marine protists and to support the implementation of management for harmful algal blooms, which are difficult to analyze using microscopy alone.

• 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.

• Korean Journal of Ecology and Environment
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• v.47 no.4
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• pp.350-357
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• 2014

Harmful Algal Bloom Alert System (HABAS) for drinking water supply is require to fast and accurate count as system monitoring of cyanobacterium occurrence and inducing a response action. We measured correlation between colony size and cell number including genus Anabaena, Aphanizomenon, Microcystis, Oscillatoria which are targeted at HABAS, deducted from standard formula, and suggested calculation method from colony size to the number of cell. We collected cyanobacteria samples at Han River (Paldang reservoir), Nakdong River (Dalseong weir, Changnyeonghaman weir) and Geum River (Gobok reservoir) from August to October, 2013. Also, we studied correlation between colony size and cell number, and calculated regression equation. As a result of correlation of harmful cyanobacteria by genus, Anabaena spp. and Aphanizomenon spp. having trichome showed high correlation coefficients more than 0.93 and Microcystis spp. having colony showed correlation coefficient of 0.76. As a result of correlation of harmful cyanobacteria by species, Anabaena crassa, Aphanizomenon flos-aquae, A. issatschenkoi, Oscillatoria curviceps, O. mougeotii having trichome showed high correlation coefficients from 0.89 to 0.96, and Microcystis aeruginosa, M. wessenbergii, M. viridis having colony showed correlation coefficients from 0.76 to 0.88. Compared with other genus Microcystis relatively showed low correlation because even species and colony size are the same, cell density and cell size are different from Microcystis strains. In this study, using calculated regression might be fast and simple method of cell counting. From now on, we need to secure additional samples, and make a decision to study about other species.