Biomonitoring the effects of urban-stream waters on the health status of pale chub (Zacco platypus): A comparative analysis of biological indexes and biomarker levels.

The objective of this study was to biomonitor the effects of potential environmental pollutants in urban-stream waters, on fish health. Pale chub (Zacco platypus), a dominant species in the Korea urban stream waters, was chosen and biomonitoring indicators for the different spatial characteristics were tailored in an urban watershed. Biological responses including biotic-somatic index as well as gonadal development phase and plasma steroids levels, and the biochemical responses, ethoxyresorufin-o-deethylase (EROD) and acetylcholinesterase (AChE) activities, were measured. No significant difference was observed in the length-weight relationship between the up-stream waters and the down-stream waters. However, changes in the gonad-somatic index (GSI) levels, plasma 17ß-estradiol (E2) levels, and mature oocyte frequencies in the female fish collected during the spawning season were observed in the down-stream waters at each monitoring site. Moreover, intersex condition (testis-ova) in the male fish in down-stream waters was recorded, even if it was just one fish. Although no significant difference was observed in the EROD and AChE activities between the up-stream waters and the down-stream waters, changes in the reproductive biomarker levels, including the GSI levels, plasma E2 levels, and gonadal maturation, lead to variable biomonitoring endpoints between the spatial different sites. These results imply that exposure to the down-stream waters can cause reproductive impairment in wild Z. platypus, individual variability in the biological responses further indicate the reproductive health was affected more by the down-stream waters than the up-stream waters. The finding from this study can provide the biomonitoring endpoint on the wild fish health in urban watershed that is crucial to the early risk assessment of its biological impacts. More multi-biomarkers studies reflecting the variation in the biological organization of wild fish and, therefore, the effects of urban-stream waters in the fish health are warranted.

Towards a multi-bioassay-based index for toxicity assessment of fluvial waters.

Despite their proven reliability for revealing 'acceptable' degrees of toxicity in waste- and reclaimed waters, bioassays are rarely used to assess the toxicity of hazardous contaminants present in natural waters. In this study, we used organisms from different trophic levels to assess the toxicity of water samples collected from four different South Korean rivers. The main objective was to develop a multi-descriptor index of toxicity for undiluted river water. The responses of six test organisms (Aliivibrio fischeri, Pseudokirchneriella subcapitata, Heterocypris incongruens, Moina macrocopa, Danio rerio and Lemna minor) after laboratory exposure to water samples were considered for this index, as well as the frequency of teratologies in diatom assemblages. Each individual test was attributed a toxicity class and score (three levels; no toxicity = 0, low toxicity = 1, confirmed toxicity = 2) based on the organism's response after exposure and a total score was calculated. The proposed index also considers the number of test organisms that received the highest toxicity score (value = 2). An overall toxicity category was then attributed to the water sample based on those two metrics: A = no toxicity, B = slight toxicity, C = moderate toxicity; D = toxicity and E = high toxicity. The susceptibility of the test organisms varied greatly and the sensitivity of their response also differed among bioassays. The combined responses of organisms from different trophic levels and with different life strategies provided multi-level diagnostic information about the intensity and the nature of contamination.

Transcriptome and computational approaches highlighting the molecular regulation of Zacco platypus induced by mesocosm exposure to common disinfectant chlorine.

Chlorine (Cl2) is a disinfectant often used in swimming pools and water treatment facilities. However, it is released into aquatic ecosystems, where it may harm non-targeted organisms. Here, we performed a mesocosm experiment exposing Zacco platypus (Z. platypus) to biocide Cl2 for 30 days (30 d) at two days' time points 15 days (15 d) and 30 d samples were collected. Here, Z. platypus was exposed to a sublethal concentration (0.1 mg/L) of Cl2, and comparative transcriptomics analyses were performed to determine their response mechanisms at the molecular level. According to RNA sequencing of the whole-body transcriptome, 860 and 1189 differentially expressed genes (DEGs) were identified from the 15 d and 30 d responses to Cl2, respectively. After enrichment analysis of GO (Gene Ontology) functions and KEGG (Kyoto Encyclopedia of Genes and Genomes) pathways, identified DEGs were demonstrated to be associated with a variety of functions, including "ion binding and transmembrane transporters". Cl2 also induced oxidative stress in Z. platypus by increasing the levels of reactive oxygen species (ROS) while decreasing the catalase (CAT) content and the levels of solute carrier family 22 member 11 (slc22a11), Caspase-8 (casp-8), inducible nitric oxide synthase (NOS2), cytosolic phospholipase A2 gamma (PLA2G4). However, Z. platypus still allows recovery during stress suspension by increasing the expression levels of solute carrier family proteins. The GO and KEGG annotation results revealed that the expression of DEGs were related to the detoxification process, immune response, and antioxidant mechanism. Additionally, protein-protein interaction networks (PPI) and cytoHubba analyses identified sixteen hub genes and their interaction. These findings elucidate the regulation of various DEGs and signaling pathways in response to Cl2 exposure, which will improve our knowledge and laid foundation for further investigation of the toxicity of Cl2 to Z. platypus.

Comparison of the Estimation Methods from Acute to Chronic Biotic Ligand Model-Based Predicted No-Effect Concentrations for Nickel in Freshwater Species.

Biotic ligand models (BLMs) and the sensitivities of indigenous species are used to assess the environmental risk considering the bioavailability of metals, such as nickel. However, the BLM-based acute-to-chronic ratio (ACR) is required if the predicted no-effect concentration (PNEC) cannot be derived from the chronic species sensitivity distribution (SSD). The applicability of the ACR approach for estimating BLM-based PNEC for nickel from acute toxicity data was evaluated in the present study. The BLM-based acute SSD for nickel was built using the sensitivities of 21 indigenous species and different taxon-specific BLMs for each taxonomic group. To predict the acute sensitivity of invertebrates, the chronic crustacean nickel BLM with pH effect term, which can account for nickel toxicity at high pH levels, was used. This was used instead of the existing acute BLM for crustacean, which has too narrow a pH range to cover the pH dependency of toxicity. The final BLM-based ACR of nickel, determined within a factor of 1.53 from the species-specific acute and chronic sensitivities of the six species, was more reliable than the typical ACR estimated within a factor of 1.84. A linear relationship (r2 = 0.95) was observed between the PNECs using BLM-based ACR and the PNECs derived from the BLM-based chronic SSD of the European Union Risk Assessment Reports. In conclusion, the BLM-based PNEC for nickel could be derived using the ACR approach, unlike when copper BLM was applied. The BLM-based ACR for nickel is the first result calculated by directly comparing acute and chronic species sensitivities, and will contribute to the application of BLM-based risk assessment in broader ecoregions. Environ Toxicol Chem 2023;42:914-927. © 2023 SETAC.

Transformation Products of Emerging Pollutants Explored Using Non-Target Screening: Perspective in the Transformation Pathway and Toxicity Mechanism-A Review.

The scientific community has increasingly focused on forming transformation products (TPs) from environmental organic pollutants. However, there is still a lot of discussion over how these TPs are generated and how harmful they are to living terrestrial or aquatic organisms. Potential transformation pathways, TP toxicity, and their mechanisms require more investigation. Non-target screening (NTS) via high-resolution mass spectrometry (HRMS) in model organisms to identify TPs and the formation mechanism on various organisms is the focus of this review. Furthermore, uptake, accumulation process, and potential toxicity with their detrimental consequences are summarized in various organisms. Finally, challenges and future research initiatives, such as performing NTS in a model organism, characterizing and quantifying TPs, and evaluating future toxicity studies on TPs, are also included in this review.

Derivation of acute copper biotic ligand model-based predicted no-effect concentrations and acute-chronic ratio.

The copper biotic ligand model (BLM) can quantitatively describe the bioavailability depending on various environmental factors and has been used to derive the predicted no-effect concentrations (PNECs). The commonly employed acute BLM tool, HydroQual, which applies the biotic ligand constants of fathead minnow in the same model structure for all taxonomic groups, estimates lower acute copper toxicity values compared to the chronic copper PNECs of the European Union Risk Assessment Reports (EU-RAR), which are based on taxon-specific model structures and biotic ligand constants for vertebrates, invertebrates, and algae. In this study, the full-BLM approach was applied using an appropriate acute BLM for each taxonomic group to derive acute HC5s (fifth percentile value in the species sensitivity distribution [SSD]) and an acute-chronic ratio for copper. Two acute BLMs for vertebrates and invertebrates were used and validated against site waters using the new method to estimate the intrinsic sensitivity for each species across different environmental conditions. To derive acute copper full BLM-based HC5s in Korean freshwater, acute toxicity tests were performed with 10 indigenous species, which were used to build the acute BLM-based SSD at each site. The final estimated acute full-BLM HC5s were higher than the EU-RAR chronic PNECs within the BLM calibration range. Furthermore, a linear relationship was observed between the acute full-BLM HC5s and the EU-RAR chronic PNECs. This linear regression function was suggested as an acute to chronic transformation function that can be applied to calculate chronic PNEC values. In conclusion, if the chronic ecotoxicity database of indigenous aquatic organisms for copper is lacking, it may be more efficient to derive chronic PNECs using an acute-chronic ratio after deriving BLM-based acute copper SSDs for indigenous species within representative taxonomic groups. This study provides a scientific foundation for the derivation of water quality criteria for copper in freshwater.

Ecological Effects of Benzyl Chloride on Different Korean Aquatic Indigenous Species Using an Artificial Stream Mesocosm Simulating a Chemical Spill.

In this study, an artificial stream mesocosm consisting of a head tank, faster-flowing riffle section, gravel section, pool section, lower-run section, and tail tank was installed to simulate a chemical spill in a river. The responses of freshwater periphyton algae, crustacea (Moina macrocopa), freshwater worm (Limnodrilus hoffmeisteri), benthic midge (Glyptotendipes tokunagai), and fish (Zacco platypus and Aphyocypris chinensis) were observed after exposure to benzyl chloride (classified as an accident preparedness substance, APS) at concentrations of 1, 2, and 4 µL/L for 22.5 h. Higher concentrations increased the inhibition (photosynthetic efficiency decrease) of periphyton algae and the mortality of M. macrocopa, whereas the reproduction of the female cladoceran decreased in the 4 µL/L treatment. Mortality of fish did not occur or was lower (≤20%) at all concentrations; however, toxic symptoms were observed for some time after chemical exposure termination and later, symptoms receded. G. tokunagai mortality increased at all concentrations except the control after seven days, and no significant toxic effects were observed in L. hoffmeisteri. The hazardous concentration of benzyl chloride was calculated as 94 µg/L. This study showed the different sensitivities of each species to benzyl chloride. The findings can assist in environmental risk assessment of APSs after chemical spills to protect Korean aquatic species.

Development of water quality criteria of ammonia for protecting aquatic life in freshwater using species sensitivity distribution method.

Ammonia is deemed one of the most important pollutants in the freshwater environment because of its highly toxic nature and ubiquity in surface water. This study thus aims to derive the criteria for ammonia in freshwater to protect aquatic life because there are no water quality criteria for ammonia in Korea. Short-term lethal tests were conducted to perform the species sensitivity distribution (SSD) method. This method is widely used in ecological risk assessment to determine the chemical concentrations to protect aquatic species. Based on the species sensitivity distribution method using Korean indigenous aquatic biota, the hazardous concentration for 5% of biological species (HC5) value calculated in this study was 44mg/L as total ammonia nitrogen (TAN). The value of the assessment factor was set at 2. Consequently, the criteria for ammonia were derived as 22mg/L at pH7 and 20°C. When the derived value was applied to the monitoring data nationwide, 0.51%, 0.09%, 0.18%, 0.20%, and 0.35% of the monitoring sites in Han River, Nakdong River, Geum River, Youngsan River, and lakes throughout the nation, respectively, exceeded this criteria. The Ministry of Environment in Korea has been considering introducing water quality standard of ammonia for protecting aquatic life. Therefore, our results can provide the basis for introducing the ammonia standard in Korea.

Stressor identification and health assessment of fish exposed to wastewater effluents in Miho Stream, South Korea.

This study evaluated the effects of an industrial wastewater treatment plant (IWTP) and a municipal wastewater treatment plant (MWTP) effluents on a variety of bioindicators ranging from biochemical, organism, and population-level responses in pale chub (Zacco platypus) and fish community structure. The Index of Biotic Integrity (IBI) indicated that the site upstream of these wastewater treatment plant discharges is in fair-good condition and downstream of the plant is in poor condition. The EROD (ethoxyresorufin-O-deethylase) activity, condition factor, and liver somatic index were significantly increased at the downstream site compared to those of the upstream site. The most significant change observed in pale chub population in the downstream site of the Miho Stream, relative to the upstream population, was the total absence of an younger age group. Stressors impacting the downstream site were identified as mostly organic or nutrient enrichment and habitat degradation associated with wastewater treatment plants. The results of causal analysis suggest that the primary causes affecting fish population in the downstream site are through both size-selective mortality caused by ammonia toxicity and recruitment failure caused by habitat degradation and reproduction problem due to an IWTP and MWTP effluents.

A new approach of Integrated Health Responses (IHR(s)) modeling for ecological risk/health assessments of an urban stream.

The objective of this study was to evaluate the ecological health of an urban stream using Integrated Health Responses (IHRs). Water chemistry analysis, habitat health, and ecotoxicity tests were conducted in the stream along with analyses of molecular/biochemical, physiological biomarkers, and population-level responses in indicator species. Chemical stresses, measured as nutrient levels, ionic content and organic matter concentrations were significantly greater (p<0.01) at the downstream than the reference site (RF). The habitat health was largely impacted in the downstream reaches and had a negative relation with the land-use pattern of % urban area. Comet assay, 7-ethoxyresorufin-O-deethylase (EROD), acetylcholinesterase (AChE), and vitellogenin (VTG) were evaluated for low-level biomarker responses on DNA/physiological conditions of target species. The multi-metric fish model (Mm-F) was used to test the community-level response in relation to chemical and physical habitat stresses. The impaired responses of separate biomarker and bioindicator at the downstream sites occurred at all organizations from molecular/biochemical level to community level. Using all biomarkers/bioindicators, the star-plot model of IHRs was developed and then the integrative health/risk assessments were conducted in the urban stream. The reduced values of IHRs occurred in the downstream sites and the impacts were attributed to effluents from wastewater treatment plants (WTPs) and industrial complex. Ecological health impairments, thus, were evident in the urban reach, and reflected the long-term community responses as well as short-term responses of molecular biomarkers. The degradation of the urban stream was mainly due to a combined effect of chemical pollution and physical habitat modifications.

Assessing effects of stress across levels of biological organization using an aquatic ecosystem health index.

Multimetric approaches typically consider only one level of biological organization to assess the effects of environmental stressors on the health of aquatic ecosystems. The present study applied integrative star-plot analysis to evaluate effects of stressors over several levels of biological organization ranging from the sub-organism to the community level at study sites subjected to different levels of contaminant stress. An aquatic ecosystem health index (AEHI), based on the sum of all the star-plot areas over these levels of biological organization, was developed to reflect an integrative and holistic assessment of stressors on ecosystem health. Star-plot areas ranged from 1.96 at a reference site to 0.79-1.08 at sites located at increasing distances downstream from a pulp mill discharge. The values of the AEHI were positively correlated with the index of biotic integrity (IBI) scores (Pearson's r=0.824). The AEHI can be used to evaluate the overall health status of aquatic systems, to identify those levels of biological organization that have the greatest response to environmental stressors, and to help identify possible causes of observed effects. Results from this study suggest that the AEHI approach can avoid false-negative responses that can occur from assessing effects at only one level of biological organization.