Spatiotemporal variation of sterols in sediment as markers of primary production and ocean sewage dumping in the southwestern East Sea (Japan Sea).

The sedimentation of organic carbon in the Ulleung Basin, in the southwestern East Sea (Japan Sea) was investigated using radiocarbon and sterols. The accumulation rates of organic carbon and the contents of brassicasterol and dinosterol were higher on the slope than in the central basin, reflecting the surface water productivity, whereas cholesterol showed similar or higher contents in the central basin. The coprostanol concentration in surface sediments reflected the dispersion of sewage dumped in this region. The vertical distribution showed that the coprostanol concentration was the highest in the top 5-cm layer near the Korea Strait, close to one of the two dumping sites. A high coprostanol concentration was also found near the coast further north, where the content peaked at ∼10 cm depth. The vertical distribution of coprostanol helped to estimate the sediment accumulation rate at sites where radiocarbon gradient was too small or the values were too variable.

Trophic assessment of red seabream (Pagrus major) as a possible bioindicator of human activities in the South Sea of Korea using stable C and N isotopes.

To assess the impact of sand mining on resource utilization by the red seabream (Pagrus major) and the trophic structure of fish assemblages two years after mining activities, we compared stable isotope ratios (δ13C and δ15N) and isotopic niches between aggregated mining and control sites in April and August 2022. Our results showed no spatial differences in the δ13C and δ15N values of red seabream between the sand mining and control sites, suggesting that the mining did not affect their dietary resources. Furthermore, the considerable overlap among fish consumers suggested that the fish food web in mining areas has trophic functions similar to those in natural habitats after mining activities. Overall, our study enhances our understanding of ecosystem conservation and the ecological-based management of coastal areas.

Fibrinous pleuritis associated with Streptococcus canis in a leopard cat (Prionailurus bengalensis euptilurus).

Leopard cat (Prionailurus bengalensis euptilurus) is a small wild cat assessed as an endangered wildlife in Korea. There have been very few reports of their diseases. Herein, we describe fibrinous pleuritis caused by Streptococcus canis infection with excessive pleural effusion, hydropericardium, mild ascites, and liver fibrosis in a leopard cat. S. canis is a commensal microflora in domestic cats and often affects the upper respiratory tract inducing chronic and severe respiratory diseases. However, there is no literature regarding the S. canis in leopard cats. Therefore, we first report fibrinous pleuritis associated with an S. canis infection in a leopard cat.

Effects of microfiber and bead microplastic exposure in the goldfish Carassius auratus: Bioaccumulation, antioxidant responses, and cell damage.

We confirmed antioxidant-related gene expression, bioaccumulation, and cell damage following exposure to various microplastics in vivo and in vitro in the goldfish Carassius auratus. Exposure of C. auratus to a 500 µm fiber-type microplastic environment (MF; 10 and 100 fibers/L) and two sizes (0.2 and 1.0 µm) of beads (MB; 10 and 100 beads/L) for 120 h increased superoxide dismutase (SOD) mRNA expression in the liver until 24 h followed by a decrease. Whereas, catalase (CAT) mRNA expression increased from 12 h to the end of the in vivo experiment. In vitro experiments were conducted with diluted microfibers (1 and 5 fibers/L) and microbeads (1 and 5 beads/L) using cultured liver cells. The results of SOD and CAT mRNA expression analysis conducted in vitro showed a tendency similar to those of experiments conducted in vivo. The H2O2 level increased in the high-concentration experimental groups compared with that in the low-concentration groups of 0.2-µm beads. In addition, the H2O2 level increased in both MF and MB groups from 12 h of exposure. Alanine aminotransferase (ALT) and aspartate aminotransferase (AST) levels in plasma were used as indicators of liver damage in fish. The ALT and AST levels increased up to 120 h after exposure. Caspase-3 (casp-3) mRNA expression was higher in the MB group than in the MF group. We visually confirmed liver casp-3 mRNA signals using in situ hybridization. The degree of DNA damage in the MF and MB high-concentration groups increased with the exposure time. The tail length and percent of DNA in the tail of the MB group were significantly higher than those of the MF group, confirming that DNA damage was greater in the MB group. Both fiber- and bead-type microplastics induced oxidative stress in C. auratus, but the bead-type induced greater stress than the fiber-type.

Micro-polystyrene plastic and benzo[α]pyrene exposure affects the endocrine system and causes physiological stress in Carassius auratus.

Microplastics, owing to their hydrophobic properties and the various chemicals used in their production, can act as carriers of persistent organic pollutants, such as polycyclic aromatic hydrocarbons (PAHs). In this study, we exposed the goldfish Carassius auratus to benzo[α]pyrene (BaP, 10 µg/L), a representative PAH, and micro-polystyrene plastic (MP; 10 and 100 beads/L), of size 1.0 µm, as a single or complex environmental stressor, and evaluated the stress response and the resulting DNA damage. The expression of CRH and ACTH mRNA in the pituitary gland and hypothalamus, of the hypothalamus-pituitary-interrenal (HPI) axis, increased significantly after 6 h of exposure. Plasma cortisol levels showed a similar trend to the expression of stress-regulating genes along the HPI axis, and a significant increase was observed in the combined exposure groups (BaP + LMP [low-concentration MP] and BaP + HMP [high-concentration MP]) compared to those in the single exposure group. H2O2 concentration and CYP1A1 and MT mRNA expression levels in the liver were significantly higher in the combined exposure groups compared with in the single exposure groups. In situ hybridization revealed a similar pattern of MT mRNA expression, and many signals were observed in the BaP + HMP group. Furthermore, the BaP + HMP group showed more DNA damage, and the degree of DNA damage increased with exposure time for all experimental groups, except for the control group. Therefore, exposure to BaP and MP alone can induce stress in goldfish; however, when a combination of both substances is provided, their synergistic effect leads to increased stress and DNA damage. MP was confirmed to be a more serious stress-inducing factor in goldfish than BaP, based on the expression levels of stress-regulating genes along the HPI axis.

Coupling and decoupling of marine stramenopiles and cyanobacteria in eutrophic coastal waters of Korea.

The spatiotemporal distribution of MASTs (MArine STramenopiles), mostly affiliated with heterotrophic protists, and their interactions with Synechococcales were investigated in an anthropogenically polluted bay of the East Sea using 18S rRNA and 16S rRNA gene sequences. The bay was characterized by strong stratification between the surface and bottom layers and cold and nutrient-rich water intrusion in summer, whereas the bay water was well mixed in winter. MAST-3, MAST-6, MAST-7, and MAST-9 were the major MAST clades, whereas the dominance of MAST-9 declined from >80 % in summer to <10 % in winter and the diversity of MAST communities increased in winter. Co-occurrence network analysis via the sparse partial least squares revealed that MAST-3 had a Synechococcales-specific interaction during the study periods but prey-specific interactions with other MAST clades were not detected. Temperature and salinity markedly influenced the relative abundance of major MAST clades. The relative abundance of MAST-3 increased at temperatures above 20 °C and salinities above 33 ‰, however, the abundance of MAST-9 decreased under the same conditions. Analysis of the metabolic functions of cyanobacteria using the FAPROTAX (Functional Annotation of Prokaryotic Taxa) indicated that the response of photosynthetic cyanobacteria to NH4+ and PO43- was significant in summer but these functions were not tightly coupled with the abundance of Synechococcales. Similarly, strong associations of MAST-3 with high temperature/salinity and Synechococcales were indicative of coupled cascading during bottom-up processes. However, other major MAST clades were likely decoupled with Synechococcales and environmental conditions in which cyanobacteria can thrive. Therefore, our results demonstrated that MAST communities can couple or decouple with environmental variables and potential prey depending on MAST clades. Collectively, our findings provide novel insights into the role of MAST communities in microbial food webs in eutrophic coastal waters.

Estuarine dam water discharge enhances summertime primary productivity near the southwestern Korean coast.

The gate opening of estuarine dams discharge a large amount of freshwater into coastal zones during the summer monsoon in northeast Asia. We investigated seasonal and spatial variations in primary productivity (PP) and community structures of phytoplankton on a downstream macrotidal flat and examined the dam discharge effects. Our harmonic analysis of the PP revealed a clear seasonality with a unimodal peak in summer, possibly driven by high ammonium loading through internal recycling. External nitrate supply from the dam-water discharge promoted PP during July-August, generating conspicuous blooms near the discharging site. Phytoplankton community was characterized by a predominance of diatoms all year round and seasonal dominances from dinoflagellates, cryptophytes, and prasinophytes in spring to chlorophytes in summer, reflecting spatiotemporal patterns in ammonium and nitrate supply. Our findings provide new insights into the summer phytoplankton bloom linked to monsoonal rainfall in the shallow coastal seas along the Northeast Asian coast.

Machine learning-guided prediction of potential engineering targets for microbial production of lycopene.

The process of designing streamlined workflows for developing microbial strains using classical methods from vast amounts of biological big data has reached its limits. With the continuous increase in the amount of biological big data, data-driven machine learning approaches are being used to overcome the limits of classical approaches for strain development. Here, machine learning-guided engineering of Deinococcus radiodurans R1 for high-yield production of lycopene was demonstrated. The multilayer perceptron models were first trained using the mRNA expression levels of the key genes along with lycopene titers and yields obtained from 17 strains. Then, the potential overexpression targets from 2,047 possible combinations were predicted by the multilayer perceptron combined with a genetic algorithm. Through the machine learning-aided fine-tuning of the predicted genes, the final-engineered LY04 strain resulted in an 8-fold increase in the lycopene production, up to 1.25 g/L from glycerol, and a 6-fold increase in the lycopene yield.

Dominance of the scleractinian coral Alveopora japonica in the barren subtidal hard bottom of high-latitude Jeju Island off the south coast of Korea assessed by high-resolution underwater images.

Coastal benthic communities in temperate regions have been influenced by climate change, including increasing sea-surface temperature. Nevertheless, scleractinian coral Alveopora japonica Eguchi, 1968, is thriving in shallow subtidal hard bottoms around Jeju Island, off the southern coast of Korea. The presence of this corals has negatively impacted subtidal kelp populations in Jeju Island. However, there is no study to document how the presence or absence of this coral relates to other benthic communities. This study investigated the benthos in three shallow subtidal sites (Shinheung (SH), Bukchon (BC), and Seongsan (SS)) in northern Jeju using underwater photography. Macro-benthic organisms appearing on a 1 × 20 m line transect installed at depths of 5, 10, and 15 m at each site were analyzed. Results showed that of the three sites investigated, A. japonica colonies were most abundant at BC, accounting for 45.9% and 72.8% of the total transect area at 10 m and 15 m, respectively. At SS, A. japonica occupied 15.3% of the total area at 15 m and less than 1% at 5 m and 10 m. The same at SH accounted for 10% of the total area at 5 m, and less than 1% at 10 m and 15 m. Dead and bleached colonies accounted for 1.2-11.5% and 1.8-5.7%, respectively, at 5, 10, and 15 m at three sites. At SS, canopy-forming brown algae Ecklonia cava and Sargassum spp. accounted for 20.2 and 24.3% of the total transect area, respectively, at 5 m depth. In contrast, the percent cover of E. cava and Sargassum spp. at SH and BC ranged from 0.1 to 1.8%, respectively. Moreover, non-geniculate coralline algae dominated the subtidal substrate at SH, ranging between 60.2 and 69% at 15 and 10 m. The low cover of A. japonica in SS (at 5 m) coincided with a high percent cover of canopy-forming brown algae. However, canopy-forming brown algae were rare at all depths at SH and BC and were dominated instead by coralline algae and the scleractinian corals. This study, by utilizing a non-destructive method, provides a baseline qualitative and quantitative information for understanding the site and depth-dependent distribution of A. japonica and algal populations, which is important to understand climate change related changes in benthic communities in Jeju and elsewhere.

Reduced forms of nitrogen control the spatial distribution of phytoplankton communities: The functional winner, dinoflagellates in an anthropogenically polluted estuary.

The effects of reduced forms of nitrogen (NH4+ and dissolved organic nitrogen (DON)) on the spatial distribution of diatoms and dinoflagellates in an estuarine-coastal water continuum were investigated from 2015 to 2019. The proportion of non-DIN in total nitrogen was utilized as an indicator of DON along with direct measurements of DON. While NO3- originated from Seomjin River, the abundant NH4+ and DON occurred from Gwangyang Bay through Namhae. Diatoms were mostly confined to the upper estuarine system and dinoflagellates dominated in the regions with high levels of NH4+ and DON. Generalized additive models also presented the different responses of diatoms and dinoflagellates to increases in NH4+ and DON. Thus, our results highlight that diatoms dominate in NO3--replete water with full access to the source and dinoflagellates take over the ecologically open niche in an anthropogenically polluted estuary with full access to reduced forms of nitrogen.

Phylogenetic diversity and spatiotemporal dynamics of bacterial and microeukaryotic plankton communities in Gwangyang Bay of the Korean Peninsula.

Nutrient dynamics function globally, flowing from rivers to the ocean (estuarine-coastal zone), and are vulnerable to climate change. Microbial habitats can be affected by marine nutrient dynamics and may provide a clue to predict microbial responses to environmental heterogeneity in estuarine-coastal zones. We surveyed surface seawater in Gwangyang Bay, a semi-enclosed estuary in Korea, from 2016 to 2018 using a metabarcoding approach with prokaryotic 16S and eukaryotic 18S rRNA genes. Bacterial and microeukaryotic communities in these waters showed distinct local communities in response to environmental heterogeneity and community transition at spatiotemporal scales in the estuarine-coastal zone. The relative abundance of prokaryotic and eukaryotic operational taxonomic units suggested a microbial trophic interaction in the Gwangyang Bay waters. We found that the community assembly process in prokaryotic communities was primarily influenced by biological interaction (immigration-emigration), whereas that in eukaryotic communities was more affected by environmental stress (habitat specificity) rather than by biotic factors. Our findings in the Gwangyang Bay waters may provide information on underlying (biotic or abiotic) factors of the assembly process in microbial communities in the estuarine-coastal zone.

Variability in blue carbon storage related to biogeochemical factors in seagrass meadows off the coast of the Korean peninsula.

Coastal vegetated habitats such as mangroves, salt marshes, and seagrasses, referred to as blue carbon ecosystems, play an important role in climate change mitigation by an effective CO2 capture from atmosphere and water columns and long-term organic carbon (Corg) storage in sediments. Although seagrass meadows are considered intense carbon sinks, information on regional variability in seagrass blue carbon stock and factors influencing its capacity still remain sparse. In the present study, seagrass blue carbon storage by measuring Corg stocks in sediments and living seagrass biomass, and carbon accumulation rates (CARs) in seagrass meadows were estimated along the Korean coast. Factors affecting variability in Corg stocks were also analyzed using partial least squares (PLS) regression and principal component analysis (PCA). Projected Corg stocks in sediment, extrapolated to a depth 1 m, exhibited substantial variability among sites, ranging from 49.91 to 125.71 Mg C ha-1. The majority of Corg (96-99%) was stored in sediments, whereas the contribution of living biomass was minor. PLS regression and PCA indicated that Corg stocks in seagrass meadows are strongly associated with sediment characteristics such as dry bulk density and water and mud content. Among seagrass traits, above- to below-ground biomass ratio was significantly related to the quantity of Corg stocks in seagrass meadows. Because of the high spatial variability in Corg stocks and CARs, local and regional differences in seagrass blue carbon storage should be considered to accurately assess the climate change mitigation potential of seagrass ecosystems.

Trophic Enrichment Factors of Carbon and Nitrogen Isotopic Ratios (Δ13C and Δ15N) in Four Marine Ciliates.

Understanding the magnitude and causes of isotopic fractionation between organisms and their dietary resources is crucial for gaining knowledge on stable isotope ecology. However, little is known regarding the diet-tissue fractionation values of marine ciliates, which play a critical role in the reconstruction of microbial food webs. In the present study, we conducted experiments on two benthic (Pseudokeronopsis pararubra and Protocruzia labiata) and two pelagic (Strombidium sulcatum and Uronemella filificum) marine ciliates, where they were fed with isotopically constant foods (Chaetoceros calcitrans and Isochrysis galbana) under laboratory culture conditions to determine their carbon and nitrogen isotopic fractionation values (Δ13C and Δ15N). The stable isotope values (δ13C and δ15N) of ciliates for all experiments rapidly increased after the initial feeding, with half-lives ranging from 6.1 to 23.0h for δ13C and from 3.1 to 24.9h for δ15N. The Δ13C and Δ15N for all ciliates represented significantly positive enrichments, with overall mean fractionations of 0.6±0.2 and 1.2±0.4, respectively. Irrespective of the dietary type, both Δ13C and Δ15N were very similar for the same ciliate species. These results suggest that Δ13C and Δ15N for marine ciliates are similar to those found in common marine organisms with very little food-dependent variation. Overall, quantifying the specific isotopic fractionation of marine ciliates is expected to provide fundamental information on the trophic transfer of carbon, nitrogen, and energy flow through the microbial pathway in marine ecosystems.

Immune-metabolic receptor GPR84 surrogate and endogenous agonists, 6-OAU and lauric acid, alter Brucella abortus 544 infection in both in vitro and in vivo systems.

Brucella abortus, one of the most important members of the genus Brucella responsible for human disease, is an intracellular pathogen capable of avoiding or interfering components of the host immune responses that are critical for its virulence. GPR84, on the other hand, is a seven-transmembrane GPCR involved in the inflammatory response and its induced expression was associated with B. abortus infection of RAW264.7 cells. Here we examined the effects of the reported GPR84 surrogate and endogenous agonists, namely 6-n-octylaminouracil (6-OAU) and lauric acid (LU), respectively in the progression of B. abortus infection in a cell and mouse models. The in vitro studies revealed the LU had bactericidal effect against Brucella starting at 24 h post-incubation. Adhesion of Brucella to RAW264.7 cells was attenuated in both 6-OAU and LU treatments. Brucella uptake was observed to be inhibited in a dose and time-dependent manner in 6-OAU but only at the highest non-cytotoxic concentration in LU-treated cells. However, survival of Brucella within the cells was reduced only in LU-treated cells. We also investigated the possible inhibitory effects of the agonist in other Gram-negative bacterium, Salmonella Typhimurium and we found that both adhesion and uptake were inhibited in 6-OAU treatment and only the intracellular survival for LU treatment. Furthermore, 6-OAU treatment reduced ERK phosphorylation and MCP-1 secretion during Brucella infection as well as reduced MALT1 protein expression and ROS production in cells without infection. LU treatment attenuated ERK and JNK phosphorylation, MCP-1 secretion and NO accumulation but increased ROS production during infection, and similar pattern with MALT1 protein expression. The in vivo studies showed that both treatments via oral route augmented resistance to Brucella infection but more pronounced with 6-AOU as observed with reduced bacterial proliferation in spleens and livers. At 7 d post-treatment and 14 d post-infection, 6-OAU-treated mice displayed reduced IFN-γ serum level. At 7 d post-infection, high serum level of MCP-1 was observed in both treatments with the addition of TNF-α in LU group. IL-6 was increased in both treatments at 14 d post-infection with higher TNF-α, MCP-1 and IL-10 in LU group. Taken together, 6-OAU and LU are potential candidates representing pharmaceutical strategy against brucellosis and possibly other intracellular pathogens or inflammatory diseases.

Trophic structures of artificial reef communities off the southern coast of the Korean peninsula as determined using stable isotope analyses.

The deployment of artificial reefs (ARs) has become a popular technique for creating new hard-bottom habitats, and for enhancing biodiversity and resource abundance for fisheries. We compared colonizing faunal assemblages and reef-associated food-web structures between ARs and nearby natural reefs (NRs) off the Korean coast using stable isotope techniques. Reef communities showed high compositional disparities in colonizing assemblages. Distinct δ13C and δ15N ranges of functional groups could be used to distinguish pelagic from benthic trophic pathways in the reef food web. The isotopic niches of entire faunal assemblages, as well as individual functional groups, overlapped between NRs and ARs, resulting in equivalency for the isotopic functional indices. Mixing model estimates for carnivorous invertebrates and fish suggested strong trophic links to reef-associated faunal prey at both reef types. Finally, these results highlight a convergence in trophic structure between ARs and NRs in accordance with functional diversity in the colonized faunal assemblages.

Formyl peptide receptor 2 (FPR2) antagonism is a potential target for the prevention of Brucella abortus 544 infection.

Here, we explore the potential role of formyl peptide receptor 2 (FPR2) during Brucella abortus infection. FPR2 manipulation affected B. abortus internalization but not its growth within macrophages. During the activation of FPR2 induced by its agonist AGP-8694, a high level of Brucella uptake was accompanied by an increase in ERK phosphorylation, while intracellular survival at 24 h postincubation was observed to be associated with slightly reduced nitrite accumulation but augmented superoxide anion production. Attenuated secretion of IL-6 and IL-10 were observed 48 h postincubation in the bone marrow-derived macrophages (BMDMs) treated with the FPR2 antagonist WRW4. An opposite pattern of bacterial uptake was observed upon treatment with the FPR2 antagonist, but no significant changes in the activation of MAPKs or the production of nitrite or superoxide anion were observed. Interestingly, AGP-8694 treatment of mice did not lead to differences in spleen or liver weight but slightly enhanced bacterial proliferation was observed in the spleen. Although the weights of the spleen or liver did not differ, WRW4 treatment led to reduced bacterial proliferation in the spleen. Furthermore, FPR2 antagonist treatment was associated with high serum levels of the proinflammatory cytokines IL-12, TNF-α, IFN-γ and MCP-1, while the production of TNF-α was inhibited in AGP-8694-treated mice. IL-6 and IL-10 levels were slightly increased in AGP-8694-treated mice at 24 h postinfection. Our findings demonstrated the contribution of FPR2 via manipulating this receptor using its reported agonist AGP-8694 and antagonist WRW4 in both in vitro and in vivo systems. Although activation of the receptor did not consistently induced Brucella infection, FPR2 inhibition may be a promising strategy to treat brucellosis in animals which encourages further investigation.

Shifting resource utilization of the lacustrine shrimp Palaemon paucidens in temperate coastal lagoons of Korea as revealed by stable isotopes.

The spatial and seasonal variations in resource use of the lacustrine shrimp Palaemon paucidens were investigated in three different Korean lagoon systems in June and October 2018 by measuring their carbon and nitrogen stable isotope ratios. P. paucidens had much higher δ13C values at the permanently open lagoon (PL) as compared to the intermittently open lagoons (ILs), revealing a disparity in resource utilization. Isotopic niches of the shrimp were relatively wider at the PL than at the ILs, suggesting a greater diversity of carbon pathways in the PL system. These results indicate that the degree of water exchange with the sea, associated with lagoon geomorphology, may be a major factor influencing resource availability for P. paucidens. Our findings suggest that the duration and degree of inlet opening may affect dietary variation at the population level, and may be one of the key components of sustainable management for coastal lagoon ecosystems.

Identifying patterns in the multitrophic community and food-web structure of a low-turbidity temperate estuarine bay.

Food web dynamics outline the ecosystem processes that regulate community structure. Challenges in the approaches used to capture topological descriptions of food webs arise due to the difficulties in collecting extensive empirical data with temporal and spatial variations in community structure and predator-prey interactions. Here, we use a Kohonen self-organizing map algorithm (as a measure of community pattern) and stable isotope-mixing models (as a measure of trophic interaction) to identify food web patterns across a low-turbidity water channel of a temperate estuarine-coastal continuum. We find a spatial difference in the patterns of community compositions between the estuarine and deep-bay channels and a seasonal difference in the plankton pattern but less in the macrobenthos and nekton communities. Dietary mixing models of co-occurring dominant taxa reveal site-specific but unchanging food web topologies and the prominent role of phytoplankton in the trophic base of pelagic and prevalent-detrital benthic pathways. Our approach provides realistic frameworks for linking key nodes from producers to predators in trophic networks.

Year-to-year variation in phytoplankton biomass in an anthropogenically polluted and complex estuary: A novel paradigm for river discharge influence.

We examined the effects of nutrient availability and turbidity on phytoplankton biomass over 9 years in Gwanyang Bay, Korea, which is an anthropogenically polluted and complex estuary. While dredging and reclamation shaped geochemical features, river discharge with low-turbidity water and sewage treatment plants contributed to nutrient loading. The replete levels of nutrients and short water-residence time suggest the inapplicability of the washout theory, whereas the presence of NH4+ suppressed the growth of phytoplankton. A reduction in the river discharge caused a concomitant decline in the loading and dilution of suspended particles. All these features led to an increase in SPM, light limitation, and NH4+ concentration. GLM estimates revealed negative effects of NH4+ and SPM on chlorophyll a over 9 years while SEM verified synergistic effects of NH4+ and SPM compared with positive effects of NO2 + NO3-. Our findings provide new insights into phytoplankton bloom dynamics in Gwangyang Bay.