• Journal of Microbiology and Biotechnology
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• 제22권2호
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• pp.248-255
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• 2012

In order to develop a protocol to quantify cyanobacteria and Microcystis simultaneously, the primers and probe were designed from the conserved regions of 16S rRNA gene sequences of cyanobacteria and Microcystis, respectively. Probe match analysis of the Ribosomal Database Project showed that the primers matched with over 97% of cyanobacterial 16S rRNA genes, indicating these can be used to amplify cyanobacteria specifically. The TaqMan probe, which is located between two primers, matched with 98.2% of sequences in genus GpXI, in which most Microcystis strains are included. The numbers of cyanobacterial genes were estimated with the emission of SYBR Green from the amplicons with two primers, whereas those of Microcystis spp. were measured from the fluorescence of CAL Fluor Gold 540 emitted by exonuclease activity of Taq DNA polymerase in amplification. It is expected that this method enhances the accuracy and reduces the time to count cyanobacteria and potential toxigenic Microcystis spp. in aquatic environmental samples.

• Environmental Engineering Research
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• 제24권3호
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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.

• 환경생물
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• 제31권2호
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• pp.113-120
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• 2013

본 연구는 초음파의 주파수별 노출 시간이 녹조현상을 일으키는 Microcystis sp.의 군체에 미치는 영향을 알아보고자 하였다. 각 조사 지점 내의 녹조 생물은 남조강(Cyanophyceae)인 Microcystis sp.가 전체 조류의 약 99.5%를 차지하였다. 초음파의 노출에 따른 녹조류의 chlorophyll-a 농도를 분석해보면 노출시간이 길어짐에 따라 점차적으로 감소하였으며, 특히 70 kHz의 주파수 영역에서 녹조 생물이 세포분열을 통한 생물량의 증가를 억제하는 데 효과적인 것으로 조사되었다. 초음파에 노출되기 전 Microcystis sp.는 단세포가 점액성 물질에 조밀하게 쌓여 있는 군체의 형태로 관찰되었으나 70 kHz 초음파에 노출된 후 Microcystis sp.의 군체의 결속이 약화되고 세포가 군체에서 떨어져 나가는 것으로 나타났다. 또한 초음파는 동 식물성 플랑크톤의 형태적 변화와 운동성에 영향을 미치지 않아 다른 수중 미생물에는 부정적인 영향을 주지 않는 것으로 확인되었다. 따라서 초음파는 Microcystis sp.의 군체와 형태적 변화를 유발하고 세포를 파괴시켜 녹조현상을 일으키는 조류의 생물량을 감소시키는 데 효과적으로 작용할 것으로 사료된다.

• Journal of Microbiology
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• 제40권4호
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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.

• 한국물환경학회지
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• 제34권2호
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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 and Biotechnology
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• 제13권4호
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• pp.544-551
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• 2003

Interactive effects of UV-B and Cd on growth, pigment, photosynthesis, and lipid peroxidation have been studied in Anabaena, Microcystis, and Nostoc; all the tested cyanobacteria showed differential sensitivity to different dosage of UV-B and Cd alone as well as in combination. Phycocyanin was severely affected by UV-B and Cd by all the strains; the degree of pigment bleaching was most pronounced in Anabaena followed by Microcystis and Nostoc. $UV-B_2+Cd_2$ produced nearly 83, 78, and 65% inhibition of phycocyanin in Anabaena, Microcystis, and Nostoc, respectively. The above treatment also significantly decreased the contents of Chl ${\alpha}$ and carotenoid. Presence of capsule in Microcystis protected the phycocyanin bleaching as compared to decapsulated cells. Laboratory-grown Microcystis revealed about 75 and 80% inhibition, following $UV-B_2+Cd_2$ treatment, respectively. in capsulated and decapsulated cells. Damage caused by Cd was more pronounced than UV-B. Inhibition of photosynthesis did occur in all the test strains, being maximum in Anabaena. PS II was the most sensitive component of the electron transport chain, showing 84, 80, and 70% inhibition in Anabaena, Microcystis, and Nostoc, respectively. As compared to control, significant lipid peroxidation (6.5-fold higher) was observed in Anabaena with $UV-B_2+Cd_2$, $^{14}C-uptake$ was more susceptible to Cd and Uv-B than oxygen-evolution. Approximately 84, 80, and 76% inhibition of $^{14}C-uptake$ was observed in Anabaena, Microcystis, and Nostoc, respectively. Similarly, $UV-B_2+Cd_2$ inhibited APT content of Anabaena by 87%. This ,study suggests that inhibition of carbon fixation was due to decreased ATP content of the test cyanobacteria by UV-B+Cd, where Anabaena was the most sensitive and Nostoc the most tolerant.

• 생태와환경
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• 제49권4호
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• pp.393-399
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• 2016

국내 주요 상수원지 (영천호, 안계호, 가창호)에서 분리한 6종의 Microcystis (M. aeruginosa, M. ichthyoblabe, M. flos-aquae, M. novacekii, M. viridis 및 M. wesenbergii)를 대상으로 microcystin 생성유전자(mcy gene) 함유률 및 생성률을 조사하였다. 조사 결과에서 M. aeruginosa는 80%(55개 균주)가 mcy gene을 함유하고 있었으며, 그 다음으로 M. ichthyoblabe는 45% (10개 균주)의 함유율을 나타내었다. 그 외에 M. flos-aquae와 M. wesenbergii는 각각 11%(4개 균주)와 33% (1개 균주)가 microcystin 생성유전자를 포함하였다. 6종의 Microcystis에서 mcy gene의 함유율은 실제 microcystin 생성율과 거의 일치하였다. 특히, 가장 많은 균주에서 mcy gene이 확인된 M. aeruginosa는 대부분 균주들(75%)이 실제로 microcystin을 생성하는 것으로 조사되었다.

• Journal of Microbiology and Biotechnology
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• 제20권7호
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• pp.1152-1155
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• 2010

In order to establish an axenic (bacteria-free) culture of Microcystis aeruginosa NIER 10039 isolated from a Korean reservoir, the culture was subjected to sequential treatment, including ultrasonication, washing, and addition of antibiotics. Three broad-spectrum antibiotics, namely, kanamycin, ampicillin, and imipenem, were applied separately in that order. Axenicity of the culture was confirmed by cultivation on bacterial media and observation under epifluorescence and scanning electron microscopes. We are the first to establish an axenic culture of a Microcystis strain isolated from Korean reservoirs and can be used in physiological and molecular studies to control toxic Microcystis blooms.

• Environmental Sciences Bulletin of The Korean Environmental Sciences Society
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• 제10권S_4호
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• pp.189-196
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• 2001

The blue green alga, Microcystis aeruginosa, was found to accumulate proline under the stressful concentration of cupric ions. The changes of proline level in Microcystis aeruginosa in response to copper(Cu) have been monitored and the function of the accumulated proline was studied with respect to its effect on Cu uptake. Exposure of Microcystis aeruginosa elevated concentrations of Cu led to accumulation of fee proline depending on the concentrations of the metal in the external medium. The greater the toxicity or accumulation of the metal, the higher the amount of proline in algal cells were found. When proline was exogenously supplied prior to Cu treatment, the absorption of Cu was markedly reduced. When exogenous proline was supplied after Cu treatment, it resulted in a remarkable desorption of the adsorbed Cu immediately after the addition of proline. Pretreatment of Microcystis aeruginosa with proline counteracted with metal-induced lipid peroxidation. The results of the present study showed a protective elect of proline on metal toxicity through inhibition of lipid peroxidation and suggested that the accumulation of proline may be related to the tolerance mechanism for dealing with Cu stress.

• Journal of Microbiology and Biotechnology
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• 제33권12호
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• pp.1615-1624
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• 2023

Microcystis blooms threaten ecosystem function and cause substantial economic losses. Microorganismbased methods, mainly using cyanobactericidal bacteria, are considered one of the most ecologically sound methods to control Microcystis blooms. This study focused on gaining genomic insights into Paucibacter aquatile DH15 that exhibited excellent cyanobactericidal effects against Microcystis. Additionally, a pan-genome analysis of the genus Paucibacter was conducted to enhance our understanding of the ecophysiological significance of this genus. Based on phylogenomic analyses, strain DH15 was classified as a member of the species Paucibacter aquatile. The genome analysis supported that strain DH15 can effectively destroy Microcystis, possibly due to the specific genes involved in the flagellar synthesis, cell wall degradation, and the production of cyanobactericidal compounds. The pan-genome analysis revealed the diversity and adaptability of the genus Paucibacter, highlighting its potential to absorb external genetic elements. Paucibacter species were anticipated to play a vital role in the ecosystem by potentially providing essential nutrients, such as vitamins B₇, B₁₂, and heme, to auxotrophic microbial groups. Overall, our findings contribute to understanding the molecular mechanisms underlying the action of cyanobactericidal bacteria against Microcystis and shed light on the ecological significance of the genus Paucibacter.