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.
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.
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.
In stream ecosystem assessment, RIVPACS, which makes a simple but clear evaluation based on macroinvertebrate community, is widely used. In this study, a preliminary study was conducted to develop a RIVPACS-type model suitable for Korean streams nationwide. Reference streams were classified into two types (upstream and downstream), and a prediction model for macroinvertebrates was developed based on each family. A model for upstream was divided into 7 (train): 3 (test), and that for downstream was made using a leave-one-out method. Variables for the models were selected by non-metric multidimensional scaling, and seven variables were chosen, including elevation, slope, annual average temperature, stream width, forest ratio in land use, riffle ratio in hydrological characteristics, and boulder ratio in substrate composition. Stream order classified 3,224 sites as upstream and downstream, and community compositions of sites were predicted. The prediction was conducted for 30 macroinvertebrate families. Expected (E) and observed fauna (O) were compared using an ASPT biotic index, which is computed by dividing the BMWPK score into the number of families in a community. EQR values (i.e. O/E) for ASPT were used to assess stream condition. Lastly, we compared EQR to BMI, an index that is commonly used in the assessment. In the results, the average observed ASPT was 4.82 (±2.04 SD) and the expected one was 6.30 (±0.79 SD), and the expected ASPT was higher than the observed one. In the comparison between EQR and BMI index, EQR generally showed a higher value than the BMI index.
The objectives of this study were to analyze the fish community structures and distribution characteristics of fisheries resources in the Osip Stream and Wangpi Stream, fishery resources protection areas. For the study, we conducted fish samplings four times in the two streams from April to October 2019. In Osip Stream and Wangpi Stream, we sampled 31 species and 29 species and the dominant species were Zacco platypus (28.6%) and Squalidus multimaculatus (41.7%), respectively. According to the life type of fish species, primary freshwater species were collected 18 species (58.1%) and 21 species (72.4%), and migration fishes were sampled 4 species (12.9%) and 6 species (20.7%) in Osip Stream and Wangpi Stream, respectively. Fisheries resources species showed high relative abundance in June, and the fish biomass was highest in October. As a results of comparing the previous studies with this study, both rivers showed high biodiversity and were found to maintain stable populations of fishes. According to analysis of tolerance guilds, the proportion of tolerant species, based on the number of individuals, was composed of 14.4% and 1.1% in Osip Stream and Wangpi Stream, respectively. This very low ratio of tolerant species confirmed that both streams have very good physiochemical environment conditions. From the above results, it was judged that the fishery resource protection areas of Osip Stream and Wangpi Stream have high ecological function and preservation value. For sustainable use and management of fishery resource protection areas of Osip Stream and Wangpi Stream, it is suggested that biological disturbance management, fish discharge projects considering environmental capacity, efforts to improve the habitat environment, and establishing a fishery resource protection areas management system were necessary as considerations.
Recently, Korean government has introduced Multi Metric Indices (MMI) using various biocommunity information for aquatic ecosystem monitoring and ecosystem health assessment at the national level. MMI is a key tool in national ecosystem health assessment programs. The MMI consists of indices that respond to different target environmental factors, including environmental disturbance (e.g. nutrients, hydrological and hydraulic situation of site etc.). We used zooplankton community information collected from Korean lakes to estimate the availability of candidate zooplankton MMI indices that can be used to assess lake ecosystem health. First, we modified the candidate indices proposed by the U.S. EPA to suit Korean conditions. The modified indices were subjected to individual index suitability analysis, correlation analysis with environmental variables, and redundancy analysis among indices, and 19 indices were finally selected. Taxonomic diversity was suggested to be an important indicator for all three taxonomic groups (cladoceran, copepod, rotifer), on the other hand, the indices using biomass for large cladocerans and copepods, while the indices using abundance were suggested for small cladocerans and rotifers.
This study compares the abundance and community structure of zooplankton organisms from pelagic regions, and considers particularly the trophic levels vs. zooplankton abundances and biomass. Zooplankton samples were collected three times from May to November 2022, at 30 temperate lakes and reservoirs, which belong to four different river basins. The total zooplankton abundance, biomass and species index were showed considerable spatial variation. The spatial pattern of rotifer abundance was similar to that of total zooplankton abundance, while there were not showed similar patterns of zooplankton biomass (μg L-1) in lentic ecosystems. The rotifer strongly dominated the zooplankton assemblage in smaller lentic system than that of larger. A total of 130 species of zooplankton were identified (83 rotifers, 34 cladocerans and 13 copepods). The total average of zooplankton abundance and biomass were 213.7±342.3 Ind. L-1 (n=129) and 1382.8±1850.4 μg L-1, respectively. Total and average of zooplankton abundance were usually dominated by the rotifers (>56.9%), while those of zooplankton biomass were dominated by the cladocerans and copepods (>73.6%) in lentic ecosystems. Considering the Trophic State Index (TSI), the factors of zooplankton abundance and biomass were included in between meso- and eutrophic states (27 lakes, 90% of all). The mean abundance and biomass of zooplankton in eutrophic systems were higher than that of meso- and hypertrophic systems. From this result, we suggest that management strategy for the lentic ecosystem water environment has to be focused more on small-sized lakes and reservoirs, in terms of zooplankton assemblages.
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.
The predator-prey interaction in freshwater ecosystems is a crucial area in the ecological study field and one of example to find such interaction is to investigate stomach contents. However, traditional method through visual inspection often induce misidentification, as it depends critically on intactness of physically visible data. In this study, we utilized Next-Generations Sequencing (NGS) technology to test the applicability stomach content analysis and overcome such limitation. NGS was applied to analyze the stomach contents of the Hemibarbus labeo, Tachysurus fulvidraco, and Plecoglossus altivelis collected in the lower part of Nakdong River. As a result, T. fulvidraco had a higher number of Animalia operational taxonomic units (OTUs) intake rate than H. labeo. At the same time, P. altivelis had higher number of Plantae OTUs intake rate than T. fulvidraco and higher Protozoa OTUs intake rate than H. labeo respectively. Therefore, NGS technology application enable to overcome traditional method’s limitation and discover hidden interspecific interaction which can further be used in appropriate habitat assessment.