Freshwater jellyfish, a type of jellyfish exclusively found in freshwater, has a limited number of species but is found globally. However, their ecology and causes of occurrence are largely unknown. Therefore, understanding the distribution of polyps, which produce the larvae of freshwater jellyfish, can provide important data for comprehending their ecology. This study aims to explore the COI gene of freshwater jellyfish using environmental DNA from the microbial film in the Miho River system. Among the 12 survey points in the Miho River watershed, genetic material of freshwater jellyfish was detected in 8 points, mainly located upstream near reservoirs. These genetic materials were identified as genes of the well-known freshwater jellyfish species, Craspedacusta sowerbii. Notably, the C. sowerbii genes found in the Miho River watershed survey points were closely related to a species previously discovered in Italy. Consequently, utilizing environmental DNA to explore the genetic traces of freshwater jellyfish enables rapid screening of areas with a high likelihood of freshwater jellyfish occurrence. This approach is deemed to provide crucial information for understanding the distribution and ecology of freshwater jellyfish in Korea.

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.

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.

The Bayesian algorithm model is a model algorithm that calculates probabilities based on input data and is mainly used for complex disasters, water quality management, the ecological structure between living things or living-non-living factors. In this study, we analyzed the main factors affected Korean Estuary Trophic Diatom Index (KETDI) change based on the Bayesian network analysis using the diatom community and physicochemical factors in the domestic estuarine aquatic ecosystem. For Bayesian analysis, estuarine diatom habitat data and estuarine aquatic diatom health (2008~2019) data were used. Data were classified into habitat, physical, chemical, and biological factors. Each data was input to the Bayesian network model (GeNIE model) and performed estuary aquatic network analysis along with the nationwide and each coast. From 2008 to 2019, a total of 625 taxa of diatoms were identified, consisting of 2 orders, 5 suborders, 18 families, 141 genera, 595 species, 29 varieties, and 1 species. Nitzschia inconspicua had the highest cumulative cell density, followed by Nitzschia palea, Pseudostaurosira elliptica and Achnanthidium minutissimum. As a result of analyzing the ecological network of diatom health assessment in the estuary ecosystem using the Bayesian network model, the biological factor was the most sensitive factor influencing the health assessment score was. In contrast, the habitat and physicochemical factors had relatively low sensitivity. The most sensitive taxa of diatoms to the assessment of estuarine aquatic health were Nitzschia inconspicua, N. fonticola, Achnanthes convergens, and Pseudostaurosira elliptica. In addition, the ratio of industrial area and cattle shed near the habitat was sensitively linked to the health assessment. The major taxa sensitive to diatom health evaluation differed according to coast. Bayesian network analysis was useful to identify major variables including diatom taxa affecting aquatic health even in complex ecological structures such as estuary ecosystems. In addition, it is possible to identify the restoration target accurately when restoring the consequently damaged estuary aquatic ecosystem.

환경유전자 (eDNA)는 다양한 환경 (수중, 토양, 대기)에 존재하는 생물체로부터 유래된 유전물질을 의미한다. eDNA는 높은 민감도, 짧은 조사시간 등 많은 장점들이 존재하며 이로 인해 생물 모니터링 및 유해생물과 멸종위기 생물을 탐색하는 분야에 다양하게 활용되고 있다. 이러한 eDNA 를 채집하기 위해서는 대상생물 및 대상유전자뿐만 아니라 현장 여과방법 및 eDNA 보존방법과 같이 매우 다양한 항목들을 고려해야 한다. 특히 환경에서 eDNA를 채집하는 방법은 eDNA 농도와 직결되는 항목으로서 적절한 채집방법을 사용하여 eDNA를 채집할 때 정확한 분석결과를 얻을 수 있다. 또한 현장에서 채집한 eDNA를 보존하고 추출하는 과정에서도 정확한 방법을 사용하였을 때 현장에 분포하는 eDNA의 농도를 정확하게 파악할 수 있다. 특히 eDNA 연구를 시작하는 연구자들에게 eDNA 분야는 초기 진입 장벽이 매우 높은 기술로서 이를 위한 기초 자료가 매우 절실하다. 본 연구에서는 본 연구는 eDNA가 수생태계를 연구하기 위한 도구로서 보다 널리 이용되며, eDNA를 이용하기 시작하는 연구자들에게 도움을 주고자 수생태계에서 eDNA를 채집하고 및 운반하는 방법과 실험실에서 eDNA를 추출하는 방법을 소개하고, 보다 간편하고 효율적인 eDNA 채집 도구와 방법을 제시하였다.

이취미 물질인 2-MIB를 합성하는 원인종에 대한 정보는 담수생태계에서 이와 관련된 환경 및 경제적 문제를 해결하는 데 필수적이다. 본 연구는 북한강 수계에서 출현하는 Pseudanabaena strain을 분리·배양하고, 16S rDNA 염기서열을 이용하여 종 수준의 동정과 2-MIB 합성 유전자 탐색을 통해 이취미 물질 발생 잠재성을 분석하였다. 북한강 본류 지역 (삼봉리, 조암면, 의암호 지역)에서 분리한 Pseudanabaena strain은 총 11개로서 NHUA201911과 NHPD201909 strain을 제외하고 단일세포의 크기는 서로 유사하였다. 그러나 16S rDNA 계통분석을 통한 유전자 염기서 열의 유연관계를 분석한 결과, 분리된 strain들은 총 5개 종 (P. cinerea, P. yagii, P. mucicola, P. galeata, P. redekei)으 로 분류되었다 (40~55% 유사도). 2-MIB를 합성하는 mibC 유전자는 P. cinerea 07 strain (NHUA202007-07)와 P. yagii (NHUA202007-08), P. redekei (NHUA201911)에서만 발견 되었으며, 가스크로마토그래피 분석에 따라 실질적인 2-MIB 합성은 P. cinerea와 P. redekei 종에서 확인되었다. 본 연구 결과는 분자생물학적 수준에서 북한강 수역에서 발생하는 Pseudanabaena속 남조류의 다양도에 대한 증거를 제공하는 연구로서 북한강 수계에서 2-MIB 생산 원인종에 대한 중요한 정보를 제공한다.

Diet and temperature are the two most critical environmental factors affecting life-history traits in insects, but the combined effects of these factors have been rarely investigated. In this study, various life-history traits were recorded from adult and larval Drosophila melanogaster fed on one of eight synthetic diets differing in protein:carbohydrate ratio (P:C=1:16, 1:8, 1:4, 1:2, 1:1, 2:1, 4:1, or 8:1) under one of six ambient temperatures (13, 18, 23, 28, 31, or 33oC). The patterns of adult and larval life-history traits expressed across 48 diet-by-temperature combinations were visualized using thin-plate spline technique and the presence of any significant linear, quadratic, and correlational effects of diet and temperature on trait expressions was analyzed using a second-order polynomial multiple regression. Life-history traits exhibited qualitatively different responses to variations in both diet and temperature, with the maximal expression of each trait being achieved at a completely divergent region of the diet-temperature fitness landscape. In adult females, for example, lifespan was maximized at P:C 1:16 under 13oC, but fecundity was maximized at P:C 4:1 under 28oC. These results provide empirical support for the emerging notion that environmental factors, such as diet and temperature, can mediate life-history trade-offs in insects.

Diets influence lifespan and reproduction in insects, but little is known how temperature modulates the impacts of diet on these two key fitness components. Here we examined the interactive effects of temperature and nutrient balance on lifespan and egg production rate in Drosophila melanogaster. Newly emerged adult D. melanogaster were allowed to feed ad libitum on one of eight chemically defined diets differing in P:C ratio (1:16, 1:8, 1:4, 1:2, 1:1, 2:1, 4:1, or 8:1) under one of six ambient temperatures (13, 18, 23, 28, 31, or 33℃). For both males and females, lifespan was longest for D. melanogaster fed on P:C 1:16 diet at 13℃ and shortened as both temperature and P:C ratio increased. As indicated by a significant temperature-by-diet interaction for lifespan, the diet effects on lifespan were more pronounced at lower temperatures than at higher temperatures. Egg production rate was maximized on P:C 4:1 diet at 28℃.

Temperature can affect the ability of insects to tolerate prolonged period of food deprivation through altering the amountof energy storage, the speed of energy expenditure, or the threshold energy storage for survival. In this study, we examinedthe mechanistic basis of the temperature-dependence of starvation resistance in Drosophila melanogaster. Starvation resistancedecreased as the temperature experienced during starvation rose from 18 to 28 ̊C. This warming-mediated decrease instarvation resistance was due to accelerated energy expenditure. However, the threshold energy storage for survival wasnot affected by starvation temperature. Exposure to warm temperatures during feeding led D. melanogaster to accumulatemore energy reserves and thus to become more starvation resistant. This study highlights the important role played bytemperature in shaping the phenotypic responses of insects to starvation.

Euiam and Paldang Reservoirs have often been facing water quality problems, such as eutrophication, algal blooms and off-flavors by treated wastewater effluent (TWE) in the North-Han and the Han River basins, but little is examined on the direct biological effect of TWE. This study tested algal growth potential (AGP) of four TWEs discharged into Euiam and Paldang Reservoirs to evaluate water fertility in September 2014 and March and September 2015. Test alga was used Anabaena circinalis isolated from Paldang Reservoir. Mean concentration of T-N and T-P in TWEs was 3,956.7 μg N L-1 and 50.8 μg P L-1, and the proportion of NO3-N and PO4-P to the total fraction was 72.1% and 40.8%, respectively. Both N and P were high in TWEs, but much higher N than P concentration indicates strong P-limitation. As a consequence, the maximum AGP was determined by PO4-P concentration (r=0.998, p<0.01). Mean AGP value was 15.4 mg dw L-1 among four effluents indicating its eutrophic condition. Due to the establishment of tertiary (advanced T-P) treatment method in the studied plants recently, P concentration was significantly decreased in TWEs compared to the years prior to 2012. However, P concentration seems to be still high enough to cause eutrophication and algal blooms. Therefore, wastewater treatment to P-free level needs to be considered if effluents are directly discharged into the drinking water resources.

This study explored spatiotemporal variability of water quality in correspondence with hydrometeorological factors in the four stations of Euiam Reservoir located in the upstream region of the North-Han River from May 2012 to December 2015. Seasonal effect was apparent in the variation of water temperature, DO, electric conductivity and TSS during the study period. Stratification in the water column was observed in the near dam site every year and vanished between August and October. Increase of nitrogen nutrients was observed when inflowing discharge was low, while phosphorus increase was distinct both during the early season with increase of inflowing discharge and the period of severe draught persistent. Duration persisting high concentration of Chl-a (>25 mg m-3: the eutrophic status criterion, OECD, 1982) was 1~2 months of the whole year in 2014~2015, while it was almost 4 months in 2013. Water quality of Euiam Reservoir appeared to be affected basically by geomorphology and source of pollutants, such as longitudinally linked instream islands and Aggregate Island, inflowing urban stream, and wastewater treatment plant discharge. While inflowing discharge from the dams upstream and outflow pattern causing water level change seem to largely govern the variability of water quality in this particular system. In the process of spatiotemporal water quality change, factors related to climate (e.g. flood, typhoon, abruptly high rainfall, scorching heat of summer), hydrology (amount of flow and water level) might be attributed to water pulse, dilution, backflow, uptake, and sedimentation. This study showed that change of water quality in Euiam Reservoir was very dynamic and suggested that its effect could be delivered to downstream (Cheongpyeong and Paldang Reservoirs) through year-round discharge for hydropower generation.

This study explored spatiotemporal variability of water quality in correspondence with hydrometeorological factors in the five stations of Paldang Reservoir located in the Han River during 4 years from May 2012 to December 2015. Variability of basic water quality factors were largely related with seasonal fluctuations of hydrology. Temperature stratification occurred in the deep dam station, and prolonged hypoxia was observed during the draught year. Nitrogen nutrients were increased with decreasing inflow in which changing pattern of NH4 reversed to NO3 by the effect of treated wastewater effluent. Phosphorus increase was manifest during the period of high inflow or severe drought. Chl-a variation was reversely related with both flow change and AGP (algal growth potential) variations. Our study demonstrated that water quality variability in Paldang Reservoir was largely attributed to both natural and operational changes of inflow and outflow (including water intake) based on major pollution source of the treated wastewater (total amount of 472×103 m3 d-1) entering to the water system from watershed. In the process of water quality variability, meteorological (e.g., flood, typhoon, abnormal rainfall, scorching heat of summer) and hydrological factors (inflow and discharge) were likely to work dynamically with nutrients pulse, dilution, absorption, concentration and sedimentation. We underline comprehensive limnological study related to hydro-meteorolology to understand short- and long-term water quality variability in river-type large reservoir and suggest the necessity of P-free wastewater treatment for the effective measure of reducing pollution level of Paldang drinking water resource.