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.

Fish require movement for feeding, spawning, growth, and evading predators, making swimming ability a critical factor in their survival. Swimming speed in fish are generally classified into sustained, prolonged, and burst speed. This study assessed the swimming capabilities of native benthic freshwater fish and provided data to inform the design and construction of fishways. Three fish species were examined: Pseudobagrus fulvidraco (family Bagridae), Tridentiger brevispinis and Rhinogobius brunneus (family Gobiidae). Measurements of total length, standard length, body weight, and body height were collected for each species, and swimming ability was evaluated using a swimming tunnel apparatus. Swimming ability was determined by measuring burst speed to calculate the critical swimming speed (Ucrit) and the relative swimming speed (TL s-1). The mean critical swimming speeds were 0.73±0.15 m s-1 for P. fulvidraco, 0.75±0.1 m s-1 for T. brevispinis, and 1.34±0.12 m s-1 for R. brunneus. Additionally, a significant correlation was found between swimming ability and total length, indicating that swimming capabilities may vary based on body type and behavior. These findings contribute essential insights into the swimming abilities of native freshwater fish, offering valuable data for improving fishway design. Further research is suggested to evaluate the swimming capabilities of a broader range of freshwater fish species to enhance fishway efficiency.

Freshwater environments are rich ecosystems that support diverse microbial communities, including members of the phylum Actinomycetota critical for nutrient cycling, organic matter decomposition, and water quality maintenance. Actinomycetota known to produce numerous bioactive secondary metabolites are valuable in biotechnology, medicine, and agriculture. Despite their significance, the diversity and distribution of Actinomycetota in freshwater habitats, especially in the Republic of Korea, are underexplored. This study aimed to report the isolation and characterization of 22 previously unrecorded bacterial species of Actinomycetota from various freshwater environments in Korea. Using standard dilution plating techniques on six different culture media, 22 bacterial strains were isolated, incubated, and characterized based on colony and cellular morphologies, Gram staining, and biochemical properties. Genomic DNA was extracted and the 16S rRNA gene was sequenced to determine species identity using the EzBioCloud service with a cutoff of 98.7% sequence similarity for classification as unreported species. These strains were phylogenetically diverse, belonging to two classes, ten orders, and eighteen genera. This study enhances our understanding of bacterial diversity in freshwater ecosystems and underscores the importance of exploring microbial diversity in underexplored habitats, potentially leading to discovery of novel bioactive compounds. Findings of this study contribute valuable insights into ecological roles and biotechnological potential of Actinomycetota in freshwater environments.

Diatoms, which are reported to have over 18,000 species worldwide and approximately 2,400 species in Korea, can be found in various environments including freshwater, seawater, and wetlands. They are particularly valuable for understanding global environmental changes throughout history due to their ability to maintain their shape for extended periods of time. Instead of collecting floating diatoms using nets, low-layer substrates such as gravel and leaves, as well as sedimentary surface layers, were gathered in order to identify attached diatom species. This is because attached diatoms demonstrate higher species diversity compared to floating diatoms. In this study, seven previously unrecorded diatoms were discovered in various domestic freshwater environments. Two species were found in reservoirs (Eunotia yanomami, Gomphonella pseudookunoi), two in parasitic cones (Eunotia carverenensis, Luticola minor), two in rivers (Cavinula maculata and Prestauroneis integra), and one (Surirella brebissonii var. kuetzingii) in a lagoon. The shapes, structures, and morphological characteristics of each diatom were identified using electron microscopy.

Zooplankton biomass is essential for understanding the quantitative structure of lake food webs and for the functional assessment of biotic interactions. In this study, we aimed to propose a biomass (dry weight) estimation method using the body length of cyclopoid copepods. These copepods play an important role as omnivores in lake zooplankton communities and contribute significantly to biomass. We validated several previously proposed estimation equations against direct measurements and compared the suitability of prosomal length versus total length of copepods to suggest a more appropriate estimation equation. After comparing the regression analysis results of various candidate equations with the actual values measured on a microbalance-using the coefficient of variation, mean absolute error, and coefficient of determination-it was determined that the Total Length-DW exponential regression equation [W=0.7775×e2.0183L; W (μg), L (mm)] could be used to calculate biomass with higher accuracy. However, considering practical issues such as the morphological similarity between species and genera of copepods and the limitations of classifying copepodid stages, we derived a general regression equation for the pooled copepod community rather than a species-specific regression equation.

In ecosystems within limited resources, interspecific competition is inevitable, often leading to the competitive exclusion of inferior species. This study aims to provide foundational information for the conservation and restoration management of Microphysogobio rapidus by evaluating species’ ecological response to biological factors within its habitat. To understand this relationship, we collected food web organisms from site where M. rapidus coexist with Microphysogobio yaluensis, a specie ecologically similar to M. rapidus, and evaluated the trophic levels (TL), isotopic niche space (INS), and the overlap of INS among fishes within the habitat using stable isotope analysis. Our analysis revealed that the M. rapidus exhibited a higher TL than M. yaluensis, with TL of 2.6 and 2.4, respectively. M. yaluensis exhibited a broad INS, significantly influencing the feeding characteristics of most fish. Conversely, M. rapidus showed a narrow INS and asymmetric feeding relationships with other species, in habitats with high competition levels. This feeding characteristics of M. rapidus indicate that the increase in competitors sharing the similar resources lead to a decrease in available resources and, consequently, is expected to result in a decrease in their density.

Captive breeding and reintroduction are crucial strategies for conserving endangered species populations. However, fish raised in predator-free environments, show a lack of recognition of predationrelated stimuli such as chemical and visual signals. It is critical to recognize chemical signals from injured conspecifics, also known as alarm signals, and the order or shape of predators to indicate the spread of predation risk in the habitat. We conducted a laboratory experiment to determine and adjust the optimal exposure period to induce appropriate anti-predator behavior response to different types of stimuli (Chemical, Visual and Chemical+Visual) for the endangered species Microphysogobio rapidus. Our results demonstrate that predator avoidance behavior varies depending on the types of stimuli and the duration of predation risk exposure. First, the results showed captive-breed M. rapidus show lack of response against conspecific alarm signal (Chemical cue) before the predation risk exposure period and tend to increase response over predation risk exposure time. Second, response to predator (visual cue) tend to peak at 48 hours cumulative exposure, but show dramatic decrease after 72 hours cumulative exposure. Finally, response to the mixed cue (Chemical+visual) tend to peak prior to the predation risk exposure period and show reduced response during subsequent exposure periods. This experiment confirms the lack of responsiveness to conspecific alarm signals in captive-bred M. rapidus and the need for an optimal nature behavior enhancement program prior to release of endangered species. Furthermore, responsiveness to predator visual signal peak at 48 hours cumulative exposure, suggest an optimal predation risk exposure period of up to 48 hours. Key words: predator cognition, captive breeding, chemical signal, visual signal, endangered

To determine the cause of the population decline in Gobiobotia naktongensis, substrate preference and burying behaviour were investigated in this study. In general, the species was shown to prefer a substrate size of 1 mm or less, depending on the flow. In addition, the burying depth varied according to the size of the fish and increased with a decrease in water temperature. Our findings showed that the main cause of the population reduction was the physical changes in the substrate structure due to the dams or barrages construction. Notably, the accumulation of silt and mud in the substrate upon the formation of an upstream lentic water region for structural construction and bed armouring caused by scouring and reduced downstream inflow of fine sediment were deterministic in the fish habitat changes, causing problems in burying. As sand substrate structure is critical for the survival and inhabitation of psammophilous species, efficient strategies should be developed with proper habitat management to reduce the anthropogenic damage

큰징거미새우(Macrobrachium rosenbergii)는 양식 시설 내에서 체색 불량 및 갑각 약화와 같은 문제로 경제적 손실을 겪고 있다. 이 종은 동물성 원료 기반의 사료로 양식되지만, 야생에서는 식물체 비중이 높은 detritus를 주로 섭취한다. 새우가 야생에서 섭취한 식물체는 기본 영양소 뿐만 아니라 체색의 재료인 카로티노이드의 공급원이기도 하다. 개나리(Forsythia koreana)는 우리나라에 널리 분포하는 꽃나무로 잎에는 황색 당근에 버금가는 양의 카로티노이드가 함유 되어 있다. 본 연구에서는 큰징거미새우에게 개나리 잎을 공급하여 체색 및 건강도에 미치는 효과를 조사하였다. 실험사료는 「배합사료 100%(대조구), CON」, 「배합사료 80%+개나리 잎 분말 20%, FP」, 「배합사료 80%+가공하지 않은 개나리 잎 20%, FL」의 세 가지였으며, 각 조건 의 사료를 평균 체중 1.1 ± 0.2 g의 어린 새우들에게 10주간 공급하였다. 실험 결과, 체색의 경 우, CON의 새우들은 일관적으로 투명한 상아색을 띠었으나, FP와 FL 새우의 경우 사육일의 경과에 따라 푸른색을 거쳐 암갈색으로 변화하였다. 생존과 성장은 CON과 FP 또는 FL 사이에 유의한 차이가 없었다. 간췌장을 조직학적으로 비교한 결과, hepatopancreatic tubule의 구성 세포 중 B cell의 vacuole 크기가 CON에 비해 FP과 FL에서 훨씬 컸다. B cell의 vacuole은 영 양소의 흡수 및 소화의 역할을 하며, 개나리 잎의 공급이 큰징거미새우의 건강에 긍정적으로 작용했을 가능성을 시사한다. 이상의 결과는 큰징거미새우의 양식에 개나리 잎을 활용하면 성장 저해 없이 체색의 개선 및 건강도의 향상을 기대할 수 있음을 보여준다.

남강댐 하류역에 위치한 사천만, 진주만, 강진만에서 가화천 방류에 따른 담수 배제의 분배 특성을 평가하기 위하여 입자추적 수치실험을 실험하였다. 가화천을 통해 3개의 방류조건(무방류, 강우시 방류, 홍수시 방류)에서 입자 1000개를 투여하여 노량수도, 대방수 도, 창선해협으로 빠져나가는 입자의 수를 비교하였다. 가화천을 통한 방류량이 늘어날수록 노량수도로의 입자 분배율이 증가하고, 대방 수도로의 분배율이 감소하는 것을 확인할 수 있었다. 즉, 평소에는 가화천 하류에 위치한 물질의 약 95%가 대방수도를 통해 빠져나가다 가, 강우에 의해 남강댐의 방류량이 증가하면서 노량수도로의 분배율이 증가하며, 홍수시에는 노량수도로의 입자 분배율이 45.5% 까지 증가하는 것을 확인할 수 있었다.

This study investigated unrecorded freshwater bacterial species in Korea. Water and sediment samples were collected from the Nakdong River basin from 2020-2022. Bacterial isolates obtained through the conventional culture method with commercial media were subjected to 16S rRNA gene sequencing to identify unrecorded bacterial species. Results of 16S rRNA gene sequencing of the bacterial isolates revealed that a total of 44 bacterial isolates shared 16S rRNA gene sequence similarities of more than 98.65%, with validly published bacterial species not reported in Korea yet. These isolates were phylogenetically assigned to 4 phyla, 7 classes, 21 orders, 33 families, and 42 genera. A total of 2, 6, 12, and 24 species belonged to phyla Bacillota, Bacteroidota, Actinomycetota, and Pseudomonadota, respectively. Here, we provide details of these 44 unrecorded bacterial species, including Gram staining, colony and cellular morphologies, biochemical properties, and phylogenetic position.