Cyanobacteria Pseudanabaena strains are known to produce 2-MIB (odorous material) in freshwater systems, thereby causing problems in water use. However, their physiological responses to environmental factors in relation with 2-MIB production is not well explored. This study was conducted to evaluate the effect of temperature on the growth and 2-MIB production of Pseudanabaena redekei. The experimental cyanobacteria strains were separated from the Uiam Reservoir (North Han River) and cultured in the BG-11 medium. Temperature was set to 10, 15, 20, 25, and 30℃ for the experiment, in the reflection of the seasonal water temperature variation in situ. For each temperature treatment, cyanobacterial biomass (Chl-a) and 2-MIB concentration (intra-cellular and extra-cellular fractions) were measured every 2 days for 18 days. Both maximal growth and total 2-MIB production of P. redekei appeared at 30℃. While intra-cellular 2-MIB contents were similar (26~29 ng L-1) regardless of treated temperatures, extra-cellular 2-MIB concentration was higher only in high temperature conditions (25~30℃), indicating that the extents of 2-MIB biosynthesis and release by P. redekei vary with temperature. The 2-MIB productivity of P. redekei was much higher in low-temperature conditions (10~15℃) than high temperature conditions (25~30℃). This study demonstrated that temperature was a critical factor contributing to 2-MIB biosynthesis and its release in cell growth (r=0.605, p<0.01). These results are important to understand the dynamics of 2-MIB in the field and thereby provide basic information for managing odorous material in drinking water resources.

Cyanobacterial resting cells, such as akinetes, are important seed cells for cyanobacteria’s early development and bloom. Due to their importance, various methods have been attempted to isolate resting cells present in the sediment. Ludox is a solution mainly used for cell separation in marine sediments, but finding an accurate method for use in freshwater is difficult. This study compared the two most commonly used Ludox methods (direct sediment treatment and sediment distilled water suspension treatment). Furthermore, we proposed a highly efficient method for isolating cyanobacterial resting cells and eDNA amplification from freshwater sediments. Most of the resting cells found in the sediment were akinete to the Nostocale and were similar to those of Dolichospermum, Cylindrospermum, and Aphanizomenon. Twenty times more akinetes were found in the conical tube column using the sediment that had no treatment than in the sample treated by suspending the sediment in distilled water. Akinete separated through Ludox were mainly spread over the upper and lower layers in the column rather than concentrated at a specific depth in the column layer. The mibC, Geo, and 16S rDNA genes were successfully amplified using the sediment directly in the sample. However, the amplification products of all genes were not found in the sample in which the sediment was suspended in distilled water. Therefore, 5 g to 10 g of sediment is used without pretreatment when isolating cyanobacterial resting cells from freshwater sediment. Cell isolation and gene amplification efficiency are high when four times the volume of Ludox is added. The Ludox treatment method presented in this study isolates cyanobacterial resting cells in freshwater sediment, and the same efficiency may not appear in other biotas. Therefore, to apply Ludox to the separation of other biotas, it is necessary to conduct a pre-experiment to determine the sediment pretreatment method and the water layer where the target organism exists.

Environmental DNA (eDNA) can exist in both intracellular and extracellular forms in natural ecosystems. When targeting harmful cyanobacteria, extracellular eDNA indicates the presence of traces of cyanobacteria, while intracellular eDNA indicates the potential for cyanobacteria to occur. However, identifying the “actual” potential for harmful cyanobacteria to occur is difficult using the existing sediment eDNA analysis method, which uses silica beads and cannot distinguish between these two forms of eDNA. This study analyzes the applicability of a density gradient centrifugation method (Ludox method) that can selectively analyze intracellular eDNA in sediment to overcome the limitations of conventional sediment eDNA analysis. PCR was used to amplify the extracted eDNA based on the two different methods, and the relative amount of gene amplification was compared using electrophoresis and Image J application. While the conventional bead beating method uses sediment as it is to extract eDNA, it is unknown whether the mic gene amplified from eDNA exists in the cyanobacterial cell or only outside of the cell. However, since the Ludox method concentrates the intracellular eDNA of the sediment through filtration and density gradient, only the mic gene present in the cyanobacteria cells could be amplified. Furthermore, the bead beating method can analyze up to 1 g of sediment at a time, whereas the Ludox method can analyze 5 g to 30 g at a time. This gram of sediments makes it possible to search for even a small amount of mic gene that cannot be searched by conventional bead beating method. In this study, the Ludox method secured sufficient intracellular gene concentration and clearly distinguished intracellular and extracellular eDNA, enabling more accurate and detailed potential analysis. By using the Ludox method for environmental RNA expression and next-generation sequencing (NGS) of harmful cyanobacteria in the sediment, it will be possible to analyze the potential more realistically.

Freshwater and brackish blue-green algae were collected at 43 freshwater and brackish sites (including lakes, ponds, swamps, streams, and rivers and estuaries) throughout South Korea from March 2017 to October 2018, and were identified using light microscopy. A total 223 taxa in freshwater and 230 taxa in brackish waters in 2017 and 274 taxa in fresh and brackish waters in 2018 were identified and among them, 20 taxa were unreported taxa of blue-green algae in Korea; The new recorded taxa were Aphanocapsa marina, Calothrix fusca f. durabilis, Calothrix littoralis, Calothrix parva, Chamaesiphon minimus, Chroococcidiopsis cubana, Chroococcidiopsis fissurarum, Coelosphaerium aerugineum, Dolichospermum mendotae, Eucapsis alpine, Gomphosphaeria cordiformis, Gomphosphaeria natans, Merismopedia danubiana, Lynbya aestuarii var. gaditana, Tolypothrix tenuis, Pseudocapsa maritima, Pseudocapsa sphaerica, Pseudophormidium tenue, Trichodesmus sp. and Woronichinia elorantae.

Targeting Microcystin (MC), which is most abundantly detected in the North-Han River water area, we analyzed the relationship between the MC biosynthesis gene (mcyA gene), cyanobacteria cell density, and MC concentration, derived an RNA-MC conversion formula, and derived the cyanobacteria. The concentration of MC present in cells was predicted. In the North-Han River waters, the mcyA gene was found mainly at downstream sites of the North-Han River after Muk-Hyeon Stream junction, and higher copy numbers were found on average than other sites. In the Uiam Lake waters upstream of the North-Han River, the mcyA gene copy number increased at the Kong-Ji Stream point, and after September, the mcyA gene copy number decreased throughout the North-Han River waters. The expression of the mcyA gene was concentrated in the short period of summer due to the spatio-temporal difference between upstream and downstream water bodies. The mcyA gene expression level was not only highly correlated with MC concentration, but also correlated with the cell density of Microcystis aeruginosa and Dolichospermum circinale, which are known to biosynthesize MC. Six conversion formulas derived based on the RNA-MC relationship showed statistical significance (p<0.05) and exhibited high correlation coefficients (r) of 0.9 or higher. The expression level of MC biosynthesis gene present in eRNA determines the synthesis of cyanotoxin substances in water, quickly quantifies gene activity, and can be fully utilized for early warning of MC development.

Biological phosphorus removal is accomplished by exposing PAO(phosphorus accumulating organisms) to anaerobic-aerobic conversion conditions. In the anaerobic condition, PAO synthesize PHB(polyhydroxybutyrate) and simultaneously hydrolysis of poly-p resulting phosphorus(Pi) release. In aerobic condition, PAO uptake phosphorus(Pi) more than they have released. In this study, cyanobacteria Synechococcus sp., which is known to be able to synthesize PHB like PAO, was exposed to anaerobic-aerobic conversion. If Synechococcus sp. can remove excess phosphorus by the same mechanism as PAO, synergistic effects can occur through photosynthesis. Moreover, Synechococcus sp. is known to be capable of synthesizing PHB using inorganic carbon as well as organic carbon, so even if the available capacity of organic carbon decreases, it was expected to show stable phosphorus removal efficiency. In 6 hours of anaerobic condition, phosphorus release occurred in both inorganic and organic carbon conditions but SPRR(specific phosphorus release rate) of both conditions was 10 mg-P/g-MLSS/day, which was significantly lower than that of PAO. When converting to aerobic conditions, SPUR(specific phosphorus uptake rate) was about 9 mg-P/g-MLSS/day in both conditions, showing a higher uptake rate than the control condition showing SPUR of 6.4 mg-P/g-MLSS/day. But there was no difference in terms of the total amount of removal. According to this study, at least, it seems to be inappropriate to apply Synechococcus sp. to luxury uptake process for phosphorus removal.

유해 남조류 Microcystis 종에 의한 녹조현상은 매년 빈번하게 일어나며, 이로 인한 수자원의 질적 변화와 먹는 물 확보에 문제가 발생하고 있다. Microcystis 종에 의한 피해를 막고자 개발된 naphthoquinone (NQ) 유도체 물질의 장점 분석을 위해 국내에서 빈번히 사용되는 황토를 이용한 microcosm 실험을 하고자 하였다. 그 결과, NQ 40, NQ 2-0 물질은 선행 연구 결과와 동일하게 유해 남조류 Microcystis 종을 선택적으로 99.9%, 99.6% 제어했으며, 식물플랑크톤 종 다양성을 증진시켰다. 그러나, 황토를 처리한 실험구는 실험 초기 일시적인 조류 제어효과를 보인 후 다시 증가하였으며, 유용 조류를 포함한 모든 식물플랑크톤에게 적용되어 다른 조류의 성장은 이루어지지 않았다. 뿐만 아니라, NQ 물질을 처리한 처리구는 비생물학적, 생물학적요인 모두 영향을 미치지 않았으며, 대조구와 유사한 경향이 관찰되었다. 따라서, 최종적으로 개량된 유해 남조류 제어물질 NQ 2-0은 높은 살조효과, 선택적 제어효과, 저독성, 자연분해에 의한 비잔류성 뿐만 아니라, 편의성 및 경제성까지 갖춘 새로운 살조물질로서 현재까지 개발된 살조물질 보다 현장 생태계 적용에 가장 적합한 친환경 녹조제어 물질이라고 판단된다.

본 연구는 2005년부터 2018년까지 14년간 주암호에서 출현한 남조류의 천이 양상을 분석하여, 남조류에 영향을 미친 주요 환경요인과의 관계를 파악하고자 연구를 수행하였다. 특히 2012년 이후 유해남조류 (Harmful algae) 출현량 급감의 원인을 파악하고자 하였다. 조사기간 동안 두 지점의 유해남조류 현존량은 주암댐: 10~24,891 cells mL-1, 신평: 13~26,043 cells mL-1의 범위로 출현하였다. 전반기 시기 두 지점의 평균 현존량은 각각 2,575 cells mL-1, 2,557 cells mL-1로, 이는 후반기 시기 평균 현존량 (42 cells mL- 1, 82 cells mL-1)에 비해 약 60배, 30배의 큰 세포수 차이를 보였다. 남조류가 상대적으로 많이 출현하는 여름 시기 (6~9월)를 대상으로 상관분석을 실시한 결과, 유해남조류는 TN, 방류량, 저수량과 양의 상관관계를 보였고, 전기전도도와는 음의 상관관계를 보였다. t-검정 결과 TN, EC, 체류시간의 유의미한 차이를 확인하였다. 특히 TN은 0.566~1.292 mg L-1의 범위로 평균 0.862 mg L-1로 보였다. 전반기, 후반기 평균 TN은 전반기 0.912 mg L-1, 후반기 0.811 mg L-1로 감소하였다. 또한 유의확률은 다소 떨어지지만, TP의 경우 평균 차이는 없으나 표준편차가 비교적 큰 차이를 보여 변동성이 큼을 알 수 있었다. 강우량이 집중된 여름 (6~9월) 시기의 전반기와 후반기 강우량 감소하는 패턴이 원인인 것으로 판단된다 (전반기 263.3 mm, 후반기 219.9 mm). 즉, 전반기에 비해 강우량은 감소 하였으며, 이로 인해 TP의 변동성이 감소되었다고 판단된다. TP의 변동성의 감소로 인해 전반기에 남조류의 luxury consumption이 가능하여 남조류의 성장률이 높았을 것으로 추측된다. 따라서 주암호 유해남조류 감소 원인은 강우량의 감소로 인한 영양염의 감소 (TN) 및 변동성의 감소 (TP)의 결과로 사료된다.

본 실험에서는 대청호에서 발생한 남조류를 대상으로 SiC(Silicon carbide) 평막의 최적 운전조건을 도출하고자 하였다. 이를 위해 원수 농도에 따른 투과플럭스, 응집제 주입 조건, Air scrubbing 조건, 역세척(Backwashing) 유량 및 시간, 여과 및 역세척 시간, 응집제 종류 및 주입 농도 등에 대해 안정적으로 운전이 가능한 최적 조건을 도출하였다. 특히, 저농도의 응집제 주입에도 음전하를 띄는 조류 입자들과 전기적으로 중화를 일으켜서 생성된 미세 플럭들이 SiC 평막의 막표면에서 투수성을 증가시킨 것으로 사료된다. 이를 통해 도출된 설계인자로 제작한 Pilot Plant를 조류 제거시 적용하고자 한다. 본 연구는 환경부의 “환경정책기반공공기술개발사업”으로 지원받은 과제입니다.

Stable isotope tracers were first applied to evaluate the Microcystis cell assimilation efficiency of bivalves, since the past identification method has been limited to tracking the changes of each chl-a, clearity, and nutrient. The filter-feeders (Sinanodonta woodiana and Unio douglasiae) were assessed under the condition of cyanobacteria (Microcystis aeruginosa) blooms through an in mesocosm experiment using 13C and 15N dual isotope tracers. Chl-a concentration in the treatment mesocosm was dramatically decreased after the beginning of the second day, ranging from 116 to 66 μg L−1. In addition, the incorporated 13C and 15N atom % in the S. woodiana bivalve showed higher values than U. douglasiae bivalves. The results demonstrate that U. douglasiae has less capacity to assimilate toxic cyanobacteria derived from diet. Our results therefore also indicate that S. woodiana can eliminate the toxin more rapidly than U. douglasiae, having a larger detoxification capacity.

Although physiological and ecological characteristics of cyanobacteria have been studied extensively for decades, unknown areas still remain greater than the already known. Recently, the development of omics techniques based on molecular biology has made it possible to view the ecosystem from a new and holistic perspective. The molecular mechanism of toxin production is being widely investigated, by comparative genomics and the transcriptomic studies. Biological interaction between bacteria and cyanobacteria is also explored: how their interactions and genetic biodiversity change depending on seasons and environmental factors, and how these interactions finally affect each component of ecosystem. Bioinformatics techniques have combined with ecoinformatics and omics data, enabling us to understand the underlying complex mechanisms of ecosystems. Particularly omics started to provide a whole picture of biological responses, occurring from all layers of hierarchical processes from DNA to metabolites. The expectation is growing further that algal blooms could be controlled more effectively in the near future. And an important insight for the successful bloom control would come from a novel blueprint drawn by omics studies.

Cyanobacteria Aphanizomenon population is widely distributed in the world, and well known as harmful algae by producing toxins and off-flavor materials, thus belonging to one of the taxa that became more interested in the field of limnoecology. In this study, the frequency, intensity, and duration of Aphanizomenon occurrence were increased with the abnormal drawdown of water level in the winter in Boryeong Reservoir, and the spatial and temporal characteristics of them are compared with each other in the perspective of hydrometeorology (1998 to 2017) and limnology (2010 to 2017). In Korea, Aphanizomenon flourished mainly in high temperature, and the appearance in the low temperature was rare in total five times. The harmful cyanobacteria Aphanizomenon was observed in the low temperature (December to February) in Boryeong Reservoir from 2014, and then reached a maximum value of 2,160 cells mL-1 in January 2017. In addition, the period exceeding 1,000 cells mL-1 at this time was more than 3 months. This was simultaneously associated with abnormal water level fluctuation in the low temperature (<10℃). The large drawdown of water level in the winter season has the potential to promote or amplify the germination and development of harmful algae. Also, subsequent water quality and ecological impacts (e.g., algal toxins and off-flavor substances) need to be considered carefully.

In this study, we investigated the correlation between the number of cyanobacterial species and the water quality factors after installation of Nakdong river weirs. Factor analyses of water quality factors were also performed. The results of correlations showed that there was no significant water quality with the cyanobacterial concentration but the correlation coefficient of nutrients increased from the upstream to the downstream. There was a significant correlation between W·T and pH as important water quality factors for the growth of cyanobacteria. In the downstream region of Nakdong river, algae were more frequently detected than in the upstream region, and the blooms of algae species seem to be influenced by the higher pH and the influx of the phosphorus from the upstream tributaries. Nutrient concentration was higher in the downstream region than in the upstream region due to the effects of tributaries Geumho river and Nam river in Nakdong river. As a result of the factor analyses, nitrogen species were the principal factors in the upper region, and phorsphorus species were the principal factors in the downstream region.

September 2014 and March 2015 at the three sites (PD-1, PD-2 and PD-3) of Lake Paldang. Nitrogen and phosphorus concentrations were higher at PD-2 than at PD-1 and PD-3. TSI (TN) values at the three field sites were in the range of 65~85, which were the level of eutrophic or hypereutrophic state. TSI (TP) also showed high values (49~68), which were the level of mesotrophic or eutrophic state. Akinete germination potential was higher at PD-2 with increased nutrient (nitrogen and phosphorus), and algal growth potential also increased with nutrient enrichment.

To evaluate the relationship between dynamics of cyanobacteria and odorous compounds, a monthly monitoring of water quality and phytoplankton were conducted at the three serial lakes (Lake Ui-am, Lake Cheong-pyeong and Lake Pal-dang) in the North Han River for 11 times from May 2014 to March 2015. In the three serial lakes, phytoplankton communities showed that seasonal changes in Bacillariophyceae- Cyanophyceae-Bacillariophyceae. Anabaena and Pseudanabaena were dominant species in August and September 2014. At the same time the odors (Geosmin, 2-MIB) were also detected with high concentration. Relationship between environmental factors and cyanobacterial abundance showed a significant correlation with Anabaena circinalis and geosmin (r=0.983, p<0.01). In the case of Pseudanabaena limnetica showed a significant correlation of total nitrogen (r=0.685, p<0.01) and NO3-N (r=0.723, p<0.01). In addition, similarly Pseudanabaena limnetica and 2-MIB (r=0.717, p<0.01) was high. The odorous compounds appeared in the North Han River water were considered to be a direct relationship with cyanobacteria.

전세계적으로 2000년대 이후로 남조류에 의한 녹조의 발생이 증가하고 있으며, 이와 관련된 환경 문제가 공중건강을 위협하고 인간 활동을 제한하고 있다. 1970년대 캐나다의 호수를 중심으로 수행된 다년간의 현장 연구를 통하여, 녹조발생의 핵심적인 영양 제한인자로 인이 제시되었고, 인 저감에 대한 수계 관리가 진행되었다. 그러나 2000년대 이후, 대형 담수수계에서 특히 Microcystis에 의한 녹조 현상에서는 인뿐만 아니라 질소가 남조류 녹조 발생에 미치는 영향이 부각되고 있다. 한국의 담수 수계에서도 이와 비슷한 남조류에 의한 위해성 녹조 번성 특징을 갖고 있으므로, 이러한 패러다임의 변화는 국내 담수수계의 영양염류 관리에서 중요한 의미를 갖는다. 최근 국제적인 관련 연구를 통하여, 위해성 남조류 번성을 막기 위해 제안된 방법은 다음과 같다. 1) 질소와 인을 함께 관리하는 전략, 2) 폐수의 수집 및 처리, 3) 호소 유입수의 사전처리, 4) 저니의 준설, 5) 체류시간의 단축, 6) 조류의 효율적 회수법, 7) 조류의 침강 및 응집 등이 제시되고 있다. 추가적으로 남조류의 생태학적 특성에 기반한 지속가능하고 통합적인 담수수계의 녹조 관리기법이 수립되어야 한다. 녹조를 유발하는 남조류는 척결되어야 할 생물체가 아니라, 담수 수계에 필수적인 미생물 구성원이기 때문이다.

국내 상수원을 대상으로 시행하고 있는 조류경보제는 남조류 발생 현황을 취 정수장 등 물관리 기관에 전파하여 대응조치를 유도하는 제도로 신속하고 정확한 남조류 계수를 필요로 한다. 따라서 조류경보제 발령 기준 대상 남조류인 Anabaena, Aphanizomenon, Microcystis, Oscillatoria 속의 군체 크기와 세포수의 상관관계를 조사하고 회귀식을 도출하여 군체 크기로 세포수를 계산할 수 있는 방법을 알아보고자 하였다. 2013년 8월부터 10월까지 남조류가 과다증식한 시기에 한강(팔당호), 낙동강(달성보, 창녕함안보) 및 금강(고복저수지)의 대표지점에서 남조류 시료를 채집하였으며, 조류경보제 발령 기준 대상 남조류 속의 종별 군체 크기와 세포수의 상관 관계를 조사하여 종 및 속별 회귀식을 산정하였다. 남조류의 속별 상관분석 결과는 사상형인 Anabaena와 Aphanizomenon의 r2값이 0.93 이상으로 높은 상관성을 보였으며 구형의 Microcystis는 0.76의 상관계수 값을 나타냈다. 종 별 상관분석 결과 사상형 남조류 Anabaena crassa, Aphanizomenon flos-aquae, A. issatschenkoi, Oscillatoria curviceps, O. mougeotii는 r2값이 0.89~0.96의 범위로 높은 상관성을 나타냈으며, 구형인 Microcystis aeruginosa, M. wessenbergii, M. viridis는 0.76~0.88의 상관계수 값을 나타냈다. 다른 속에 비해 상대적으로 Microcystis의 상관성이 낮게 나타난 이유는 동일한 종, 동일한 크기의 군체라도 Microcystis strain에 따라 점액질 내의 세포 밀집 정도와 세포 크기에 차이가 있기 때문이다. 본 연구 결과 도출한 회귀식을 이용하여 군체 크기 측정값을 세포수로 환산하는 방법이 기존의 세포 계수법과 비교할 때 신속하고 간편할 것으로 보인다. 향후 남조류 종별 더 정확한 회귀식을 도출하기 위해서는 많은 시료수 확보와 더불어 다른 종들에 대한 조사 연구가 진행되어야 할 것이다.

Akinete germination may be a starting point of some akinete-producing cyanobacterial blooms in eutrophic freshwater systems. This study examined germinability of akinete of a cyanobacterium Anabaena circinalis isolated from the sediment of the North Han River (Cheongpyung Dam), Korea, under several environmental factors such as temperature (5∼25℃), light (0∼100μmol photons m-2 s-1), nutrients (nitrogen and phosphorus) and pH (5~12). The high germination rate appeared at high temperature: >55% at 25℃, followed by 15% at 15℃, 10℃ and 10% at 5℃. Low light intensity was favorable for akinete germination. Over 45% of germination occurred at low light intensities (5, 15 and 30μmol photons m-2 s-1), while less than 10% of germination occurred at both 50 and 100μmol photons m-2 s-1. No germination occurred in the dark condition. Akinete germination rate increased with nutrient (phosphorus and nitrogen) enrichment, and nitrogen addition showed greater effect on the germination compared to phosphorus addition. Akinetes germinated well at neutral or slightly alkaline pH condition (pH 7 and pH 8: >55%), but no germination was observed at pH 11~12. The present study demonstrates some favorable ambient conditions of Anabaena circinalis germination, which could provide useful information to study the germination conditions of other Anabaena species or akinete-forming algae and predict its bloom in eutrophic freshwaters.

남조류는 겨울철에 휴면포자의 형태로 퇴적층에서 존재하다가 적합한 환경조건이 되었을 때 다시 영양세포로 전환된다. 이렇게 전환된 영양세포는 성장에 적합한 환경이 되었을 때 대량증식 하게 되어 수체 내에서 우점한다. 이전까지의 국내 대부분의 연구는 수체 내 영양세포의 변동과 환경인자와의 관계 또는 영양세포에 대한 생리생태적 연구부분에 국한되었다. 하지만, 휴면포자에서 영양세포로 진행되는 부분에 대한 연구가 조류대량증식의 궁극적인 기작이 되므로 휴면포자에 대한 연구는 조류의 생활사를 이해하는 데에도 매우 중요하다. 또한 다른 조류들간의 변동 및 천이의 연구를 위해서도 필수적이다.