This study quantitatively analyzed the ecological niche breadth (ENB) of 27 freshwater mollusk species (11,406 occurrence records) in Korea based on macroenvironmental gradients, including climate, topography, and land cover. We combined Levins’ index values (B: niche breadth, BA: equitability) with optimum-tolerance analysis (optimum value and tolerance range, t95-t05) to determine species-specific habitat strategies and environmental responses. The analysis revealed a distinct dichotomy in the habitat strategies of Korean freshwater mollusks, separating cold-water, lowdisturbance types in upstream areas from warm-water, disturbance-tolerant types in downstream areas, mainly structured by thermal and land-cover gradients. Species, such as Semisulcospira libertina, Radix auricularia, and the alien species Physa acuta, were identified as generalists, showing high B values but low BA values, suggesting that they are capable of inhabiting diverse environments but concentrate their presence in certain environmental conditions. Conversely, Koreoleptoxis nodifila and Clithon retropictum were classified as specialists with low B values and intermediate to high BA values, indicating stable and equitable distribution within restricted environmental limits. Urbanization appeared to be an influential limiting factor, showing low BA values for most species, whereas agricultural areas showed higher BA values, suggesting a comparatively broader habitat possibility. Notably, K. nodifila, a critically endangered and endemic species, showed optimal habitat conditions of low temperature (10.65°C), low urbanization (4.82%), and high forest cover (57.34%), confirming its status as a cold-water indicator species adapted to forested upstream systems. The study findings provide a quantitative framework to assess the environmental tolerance, vulnerability, and conservation priorities of Korean freshwater mollusks.

Freshwater ecosystems support biodiversity and provide essential ecosystem services. In Korea, the Water Environment Information System monitors these ecosystems using separate biological and physicochemical indicators. Complex interactions occur among diverse biological taxa and physicochemical conditions. Thus, integrating heterogeneous monitoring data is crucial for accurately assessing ecosystem health. However, differences in data characteristics between the indicators present significant integration challenges. Given the scale and heterogeneity of the monitoring data, advanced analytical techniques are necessary to detect interactions among variables. This study aimed to identify key correlations among biological and physicochemical indicators by clustering similar variables and removing noise using the Hierarchical Density-Based Spatial Clustering of Applications with Noise (HDBSCAN) algorithm, followed by Spearman’s rank correlation coefficient and maximal information coefficient (MIC) analyses. HDBSCAN effectively eliminated noise indicators and grouped biological and physicochemical indicators into clusters based on shared characteristics, thereby enhancing the interpretability of the correlation analysis. Spearman analysis showed strong associations among biological indicators, particularly among species with similar ecological traits. MIC analysis further detected nonlinear associations between ecological assessment indices and specific biological species, which also reflected similar ecological characteristics. These findings are significant in that the comprehensive analysis of existing monitoring data revealed relationships within biological and physicochemical indicators while preserving the original purpose and function of each monitoring network. This study is expected to serve as a foundational resource for freshwater environmental monitoring and the development of effective management strategies.

From 2017 to 2024, we surveyed 43 diverse aquatic habitats in South Korea, leading to the identification of 18 cyanobacterial taxa that are newly recorded for the country, found across eight sites (about 18% of the surveyed locations). These taxa exhibit a wide range of morphological forms, including unicellular, colonial, filamentous, and heterocytous types, and belong to various orders such as Chroococcales, Synechococcales, Nostocales, and Stigonematales. Notably, this study provides a provisional record of Gomphosphaeria aponina in Korea, correcting its previous misidentification as G. natans. We also documented Dolichospermum compactum, a species that has been genetically reclassified. Additionally, we identified species with the potential to cause harmful algal blooms (HABs), such as Microcystis botrys and Gloeotrichia aurantiaca, which are crucial for domestic water quality monitoring. Currently, only 414 cyanobacterial taxa are recorded in Korea, representing less than 8% of the estimated global total of approximately 5,300 species. This significant gap underscores the considerable unrecorded diversity within Korean aquatic ecosystems. These findings substantially enhance the national cyanobacterial checklist and underscore the need for ongoing monitoring in understudied aquatic environments. They also highlight the importance of integrating classical morphological and ecological observations with advanced molecular methods. This polyphasic approach can accurately detect cryptic diversity and support robust ecological assessments. Overall, this comprehensive floristic expansion offers valuable baseline data for biodiversity inventories, ecological monitoring, and the development of microbial resources within Korean aquatic environments.

This study investigated the effects of elevated salinity on the growth, morphology, and biochemical composition of two freshwater microalgae species: Chlorella thermophila (a green alga) and Anabaena variabilis (a cyanobacterium). The goal was to understand their adaptive mechanisms under saline stress and evaluate their potential for biofuel production. These species were chosen for their ecological significance and contrasting cellular structures (unicellular versus filamentous), which provide complementary insights into salinity tolerance among freshwater microalgae. Cultures were maintained in both standard BG11 medium and artificial seawater medium (SWM) under controlled light intensity (200 μmol photons m-2 s-1) and pH (7.5). Over a cultivation period of 10-20 days, we quantified key parameters, including cell size, volume, chlorophyll a, protein, lipid content, and fatty acid profiles, using microscopy (ImageJ), spectrophotometry, FTIR, and GC-MS analysis. Both species showed increased cell volume and lipid accumulation in SWM, with C. thermophila experiencing a dramatic volume increase from 5.83 μm3 to 54.76 μm3) and a 6.8% rise in lipid productivity. Fatty acid profiling identified distinct fatty acids, such as palmitoleic and pantetheic acids, in C. thermophila cultivated in SWM. These specific fatty acids may indicate adaptive strategies for osmoregulation or metabolic shifts toward energy storage under salinity-induced stress. These findings highlight the potential of salinity-driven modulation of microalgal metabolism to enhance biofuel precursors, offering a sustainable approach for biomass production in saline environments with reduced reliance on freshwater.

The genus Pleurastrum is a coccoid green alga comprising 10 species worldwide. Pleurastrum exhibits simple morphology and high polymorphism, which complicates the understanding of its diversity. We examined the morphological and ultrastructural characteristics of Pleurastrum using light, confocal, and transmission electron microscopy. Additionally, we performed phylogenetic analysis based on multigene sequences (nuclear SSU rDNA, 5.8S, internal transcribed spacer (ITS2) region, and plastid rbcL and tufA genes) from Pleurastrum strains to report two previously unrecorded freshwater species (Pleurastrum insigne and Pleurastrum microstigmatum) in Korea. The vegetative cells were predominantly spherical, with a few being ellipsoidal, and each cell contained a chloroplast with one pyrenoid. The sporangia produced several daughter cells, while the biflagellate zoospores were ellipsoidal and motile. Phylogenetic analysis confirmed that P. insigne and P. microstigmatum form well-supported monophyletic clades. Analysis of ITS2 secondary structures revealed similar patterns, with several differences in nucleotide sequences and insertions between the two species. The findings of this study expand the known distribution of Pleurastrum and enhance our understanding of its species diversity in Korea.

Freshwater bivalves contribute to key ecological functions in lake ecosystems, yet their cryptic and benthic lifestyles often hinder detection through conventional surveys. In this study, we applied environmental DNA (eDNA) metabarcoding to assess the diversity and distribution of unionid bivalves in six lakes across Republic of Korea. Water samples were collected from three sampling strategies-Center Surface, Center Mix, and Waterside Surface-and processed using 16S rDNA-targeted primers followed by high-throughput sequencing. A total of four unionid species (Cristaria plicata, Sinanodonta lauta, Unio (Nodularia) douglasiae, and Anodonta woodiana) were detected across 18 sampling points. Notably, eDNA successfully identified unionid presence in all lakes, even where conventional surveys failed to observe individuals. Among the sampling strategies, Center Mix exhibited the highest values for Shannon and Simpson indices as well as ASV richness. Waterside Surface samples generally showed lower diversity and detection frequency. A Venn diagram of ASV occurrences revealed three ASVs shared across all sampling strategies and one unique ASV found only in Center Mix. These results indicate that sampling location significantly affects detection sensitivity and diversity representation in eDNA-based bivalve monitoring. Combined application of Center Mix and Center Surface strategies may enhance both detection efficiency and species diversity coverage in lentic environments.

Environmental DNA (eDNA) analysis has emerged as a powerful tool for biodiversity monitoring due to its efficiency, standardization potential, and cost-effectiveness. We evaluated the applicability of eDNAbased zooplankton monitoring in Korean lakes by comparing three DNA methods-eDNA, iDNA, and eiDNA-with traditional microscopy. Sampling was conducted in six lakes with varying conditions. eDNA was obtained from lake water, iDNA from unpreserved zooplankton incubated in water, and eiDNA from zooplankton incubated in ethanol. DNA metabarcoding detected more taxa than microscopy, but dominant taxa overlapped, mainly Daphnia. While DNA methods showed higher richness, Simpson and Shannon indices were higher in microscopy, reflecting differences in quantification methods. These discrepancies reflect methodological differences in how taxa are quantified and suggest that DNA-based approaches may overrepresent certain groups in richness estimates. In addition, false negatives were observed for several common rotifer species (e.g., Keratella, Polyarthra), likely due to incomplete reference databases and high intraspecific genetic diversity. Conversely, some taxa detected only by DNA-particularly small-bodied or rare crustaceans-may represent false positives relative to microscopy. These findings emphasize the importance of improving reference libraries and interpreting DNA results with caution, while also supporting the utility of DNA-based methods as complementary tools in zooplankton monitoring and national biodiversity assessments.

Environmental DNA (eDNA) has emerged as a promising tool for aquatic biodiversity monitoring, yet its collection in lentic ecosystems remains technically constrained by filtration capacity and field logistics. In this study, we applied a novel eDNA concentration system, QuickConcTM, to evaluate freshwater mussel diversity in lakes, and compared its performance with the conventional GF/F filtration method. Water samples were collected from four reservoirs at surface, mid, bottom, and waterside layers, and processed using both filtration techniques. Metabarcoding of mitochondrial 16S rDNA revealed that QuickConcTM captured a higher average number of amplicon sequence variants (ASVs) and exhibited greater species richness and diversity indices (Shannon and Simpson), although the differences were not statistically significant. QuickConcTM samples showed a greater capacity to detect rare taxa and to recover higher ASV richness in certain cases, suggesting its potential to enhance biodiversity resolution. Species composition remained consistent across methods, with Cristaria plicata and Sinanodonta lauta being dominant in both cases. However, slight spatial variations in species assemblages were observed between center and waterside sampling points, highlighting the influence of habitat heterogeneity on eDNA distribution. Overall, our results demonstrate that the QuickConcTM system offers a practical and efficient alternative to traditional filtration methods for eDNA-based freshwater mussel monitoring, particularly in environments with high suspended solids. The findings underline the need for adaptive sampling strategies that consider both methodological and ecological factors when designing eDNA surveys in lentic ecosystems.

Freshwater environments serve as crucial habitats for diverse microorganisms, playing essential roles alongside plants and animals within the ecosystem. Bacteria in these environments are particularly important for maintaining ecosystem functions. Among them, members of the phylum Pseudomonadota are known to be involved in processes such as nitrogen fixation, ammonia oxidation, and biofilm formation. Pseudomonadota comprises six classes: Acidithiobacillia, Alphaproteobacteria, Betaproteobacteria, Gammaproteobacteria, Hydrogenophilia, and Zetaproteobacteria. This study reports 43 previously unrecorded species belonging to the phylum Pseudomonadota. They were isolated from various freshwater habitats in Republic of Korea. These strains were isolated and cultured using nine different standard media, with classification based on Gram staining, cell morphology, and biochemical characteristics. Species identity was determined using 16S rRNA gene sequencing, with a similarity threshold of 98.7% for designation as unreported species. Phylogenetic analysis revealed that these 43 strains were distributed across three classes, 13 orders, 28 families, and 39 genera. This research emphasizes microbial diversity in freshwater ecosystems and provides valuable insights into ecological roles and potential novel functions of unreported bacterial species in Korea.

As part of the 2024 research initiative, “Investigation and Discovery of Prokaryotes in Freshwater Systems,” samples were collected from diverse freshwater habitats, including both water and soil environments. Approximately 2,000 bacterial strains were isolated as single colonies and identified through 16S rRNA gene sequence analysis. Among these, 38 strains shared ≥98.7% 16S rRNA gene sequence similarities with those of known bacterial species not previously reported in Korea. These strains were thus categorized as newly recorded bacterial species in Korea. These 38 bacterial strains displayed significant phylogenetic diversities, spanning 2 phyla, 4 classes, 15 orders, 24 families, and 34 genera. These unrecorded species were classified into the following classes: Actinomycetia (with genera including Microcella, Conyzicola, Curtobacterium, Leucobacter, Microbacterium, Frigoribacterium, Lysinibacter, Streptomyces, Nonomuraea, Actinocorallia, Ruania, and Actinoplanes), Alphaproteobacteria (Paracoccus, Youngimonas, Loktanella, Corticibacterium, Neorhizobium, Onobrychidicola, Ferranicluibacter, Aureimonas, Asticcacaulis, and Novosphingobium), Betaproteobacteria (Rhodoferax, Rugamonas, and Cupriavidus), and Gammaproteobacteria (Rheinheimera, Shewanella, Kosakonia, Leclercia, Hafnia, Yersinia, Pseudomonas, Lysobacter, and Acinetobacter ). Further characterization included assessment of Gram reaction, colony and cell morphology, biochemical properties, and phylogenetic relationships. This report presents detailed phylogenetic and phenotypic characteristics of these bacterial species.

During an investigation of indigenous prokaryotic species in the Republic of Korea, 17 bacterial strains belonging to the phylum Bacteroidota and 4 strains belonging to the phylum Bacillota were isolated from freshwater environments. These isolated bacterial strains formed a robust phylogenetic clade with type strains of the closest related bacterial species, with 16S rRNA gene sequences sharing similarities of higher than 98.7%. To date, there have been no official report of these 21 isolates in the Republic of Korea. At the genus level, these unreported species were affiliated with Mucilaginibacter and Pedobacter of class Sphingobacteriia, Flavobacterium and Gillisia of class Flavobacteriia, Hymenobacter of class Cytophagia, and Paenibacillus and Planococcus of class Bacilli. These species were further examined by performing Gram staining, analyzing their colonies and cell morphologies, and determining their basic biochemical characteristics and phylogenetic positions. Detailed descriptions of the 21 previously unreported species are provided.

Microalgae are efficient fatty acid producers owing to their high photosynthetic activities. They can act as sources of biofuel, feed, and various bioactive compounds. This study aimed to determine optimal culture conditions, including culture medium, temperature, and light intensity, to enhance the biomass and fatty acid content of the indigenous freshwater microalga, Tetradesmus obliquus. Evaluation using a high-throughput photobioreactor revealed that the optimal culture temperature and light intensity were 25°C and 300 μmol m-2 s-1, respectively. Additionally, we optimized components (N, P, and Mg) of the BG-11 medium to enhance the microalgal biomass. Modified BG-11 medium increased the T. obliquus biomass by 37% compared to the standard BG-11 medium. Subsequently, the culture medium was replaced with N- and P-depleted media to determine the abiotic stress factor that could increase the cellular fatty acid content. Notably, fatty acid content was significantly increased from 8.5% up to 14.6% on day 7 of culture in N-deficient (N-P+ and N-P-) media. Sequential optimization effectively increased the biomass by 83% and fatty acid content by >76% in T. obliquus. Our optimization method can be used to enhance the biomass and fatty acid contents of various other microalgae.

Biodiversity encompasses species diversity, which includes species richness and species evenness. High species diversity is known to contribute to community stability and the potential for maintaining healthy ecosystem functioning. However, the most commonly used species diversity indices have some limitations, as they require species-specific abundance data for each community. In contrast, phylogenetic diversity measures the evolutionary distances between species within a community, reflecting ecological and/or evolutionary divergences and niche differences, without requiring abundance data. This study assessed biodiversity by calculating phylogenetic diversity indices for freshwater fishes (19 species) and aquatic insects (49 species) at three sites within Seoraksan and Odaesan National Parks. The aquatic insects studied belong to the EPT group (Ephemeroptera, Plecoptera, and Trichoptera), which are widely used as bioindicators of aquatic ecosystems. Two mitochondrial DNA genes were used as molecular markers: COI and cyt b for fish, and COI and 16S rRNA for the EPT - group. Overall, Odaesan National Park exhibited higher phylogenetic diversity in both fishes and aquatic insects compared to Seoraksan National Park, although this difference was not statistically significant. The highest phylogenetic and species diversity were observed for fish at the OD2 site (Woljeong District) and for the EPT group at the OD1 (Gyebangsan) and SA2 (Jangsudae) sites. Correlation analysis revealed that phylogenetic diversity indices were more positively associated with species richness than species diversity indices. This study serves as a pilot project for establishing standardized methods for assessing biodiversity in national park habitats using phylogenetic diversity. It also aims to inform various policies, such as the development of biodiversity assessment systems and the prioritization of protected areas within national parks.