Integrated approach for the isotope trophic position of black-tailed gull (Larus crassirostris) eggs over a decade: Combining stable isotopes of amino acids and fatty acids composition.

Recently, compound-specific isotope analysis (CSIA) using the amino acid nitrogen stable isotope ratio (δ15NAAs) has been widely used for accurate estimation of trophic position (TP). In addition, a quantitative fatty acid signature analysis (QFASA) offers insights into diet sources. In this study, we used these techniques to estimate the TP for seabirds that rely on diverse food sources across multiple ecosystems. This allows for the proper combination of factors used in TP calculation which are different for each ecosystem. The approach involved the application of a multi-mixing trophic discrimination factor (TDF) and mixing ß which is a Δδ15N between trophic and source amino acid of primary producer. Since the black-tailed gulls (BTGs) are income-breeding seabirds, which rely on energy sources obtained around their breeding sites, they and their eggs could be useful bioindicators for environmental monitoring. However, the ecological properties of BTGs such as habitats, diets, and TP are not well known due to their large migration range for wintering or breeding and their feeding habits on both aquatic and terrestrial prey. In this study, the eggs were used for estimating TP and for predicting TP of mother birds to overcome difficulties such as capturing birds and collecting non-invasive tissue samples. Eggs, sampled over a decade from three Korean islands, showed spatial differences in diet origin. Considering both the food chain and physiology of BTG, the TP of eggs was estimated to be 3.3-4.0. Notably, the TP was significantly higher at site H (3.8 ± 0.1) than at site B (3.5 ± 0.2), which indicated a higher contribution of marine diet as confirmed by QFASA. Using a reproductive shift of δ15NAAs, the TP of the mother birds was predicted to be 3.6-4.3, positioning them as the top predator in the food web. The advanced integration of multiple approaches provides valuable insights into bird ecology.

New insight into biomagnification factor of mercury based on food web structure using stable isotopes of amino acids.

Although many attempts have been carried out to elaborate trophic magnification factor (TMF) and biomagnification factor (BMF), such as normalizing the concentration of pollutants and averaging diet sources, the uncertainty of the indexes still need to be improved to assess the bioaccumulation of pollutants. This study first suggests an improved BMF (i.e., BMF') applied to mercury bioaccumulation in freshwater fish from four sites before and after rainfall. The diet source and TP of each fish were identified using nitrogen stable isotope of amino acids (δ15NAAs) combined with bulk carbon stable isotope (δ13C). The BMF' was calculated normalizing with TP and diet contributions derived from MixSIAR. The BMF' values (1.3-27.2 and 1.2-27.8), which are representative of the entire food web, were generally higher than TMF (1.5-13.9 and 1.5-14.5) for both total mercury and methyl mercury, respectively. The BMF' implying actual mercury transfer pathway is more reliable index than relatively underestimated TMF for risk assessment. The ecological approach for BMF calculations provides novel insight into the behavior and trophic transfer of pollutants like mercury.

Microplastics disrupt energy metabolism in the brackish water flea Diaphanosoma celebensis.

Energy metabolism is crucial for normal biological processes, such as growth, development, and reproduction. Microplastics disrupt energy homeostasis by modulating the digestive capacity and contents of energy reserves to overcome stress. This study investigated the modulation of digestive enzyme activity and energy reserves in the brackish water flea Diaphanosoma celebensis exposed to polystyrene (PS) beads (0.05-, 0.5-, 6-µm) for 48 h, and examined transcriptional changes in digestive enzyme-coding genes and AMP-activated protein kinase (AMPK) signaling pathway genes. PS particle size differentially modulated digestive enzyme activity, energy molecule content (glycogen, protein, and lipids), and metabolism-related gene expression. In particular, the 0.5-µm PS had the most significant effect on digestive enzyme activity. In contrast, the 0.05-µm PS caused significant metabolic disorder following a decrease in total energy budget (Ea). These findings suggest that PS beads can modulate energy metabolism through different modes depending on the bead size.

Effect of heavy metals on the energy metabolism in the brackish water flea Diaphanosoma celebensis.

Heavy metals such as lead (Pb), cadmium (Cd), and arsenic (As) are of great concern in aquatic ecosystems because of their global distribution, persistence, and biomagnification via the food web. They can induce the expression of cellular protective systems (e.g., detoxification enzymes and antioxidant enzymes) to protect organisms from oxidative stress, which is a high-energy-consuming process. Thus, energy reserves (e.g., glycogen, lipids, and proteins) are utilized to maintain metabolic homeostasis. Although a few studies have suggested that heavy metal stress can modulate the metabolic cycle in crustaceans, information on changes in energy metabolism under metal pollution remains lacking in planktonic crustaceans. In the present study, the activity of digestive enzymes (amylase, trypsin, and lipase) and the contents of energy storage molecules (glycogen, lipid, and protein) were examined in the brackish water flea Diaphanosoma celebensis exposed to Cd, Pb, and As for 48 h. Transcriptional modulation of the three AMP-activated protein kinase (AMPK) and metabolic pathway-related genes was further investigated. Amylase activity was highly increased in all heavy metal-exposed groups, whereas trypsin activity was reduced in Cd- and As-exposed groups. While glycogen content was increased in all exposed groups in a concentration-dependent manner, lipid content was reduced at higher concentrations of heavy metals. The expression of AMPKs and metabolic pathway-related genes was distinct among heavy metals. In particular, Cd activated the transcription of AMPK-, glucose/lipid metabolism-, and protein synthesis-related genes. Our findings indicate that Cd can disrupt energy metabolism, and may be a potent metabolic toxicant in D. celebensis. This study provides insights into the molecular mode of action of heavy metal pollution on the energy metabolism in planktonic crustaceans.

Stable Isotope Analysis of Residual Pesticides via High Performance Liquid Chromatography and Elemental Analyzer-Isotope Ratio Mass Spectrometry.

To broaden the range of measurable pesticides for stable isotope analysis (SIA), we tested whether SIA of the anthranilic diamides cyantraniliprole (CYN) and chlorantraniliprole (CHL) can be achieved under elemental analyzer/isotope ratio mass spectrometry with compound purification in high-performance liquid chromatography (HPLC). Using this method, carbon isotope compositions were measured in pesticide residues extracted from plants (lettuce) grown indoors in potting soil that were treated with 500 mg/kg CHL and 250 mg/kg CYN and were followed up for 45 days. Our results show that the CYN and CHL standard materials did not have significant isotope differences before and after clean-up processing in HPLC. Further, when applied to the CYN product and CHL product in soil, stable isotope differences between the soil and plant were observed at <1.0‱ throughout the incubation period. There was a slight increase in the variability of pesticide isotope ratio detected with longer-term incubation (CHL, on average 1.5‱). Overall, we measured the carbon isotope ratio of target pesticides from HPLC fraction as the purification and pre-concentration step for environmental and biological samples. Such negligible isotopic differences in pesticide residues in soils and plants 45 days after application confirmed the potential of CSIA to quantify pesticide behavior in environments.

Amino acid nitrogen and carbon isotope data: Potential and implications for ecological studies.

Explaining food web dynamics, stability, and functioning depend substantially on understanding of feeding relations within a community. Bulk stable isotope ratios (SIRs) in natural abundance are well-established tools to express direct and indirect feeding relations as continuous variables across time and space. Along with bulk SIRs, the SIRs of individual amino acids (AAs) are now emerging as a promising and complementary method to characterize the flow and transformation of resources across a diversity of organisms, from microbial domains to macroscopic consumers. This significant AA-SIR capacity is based on empirical evidence that a consumer's SIR, specific to an individual AA, reflects its diet SIR coupled with a certain degree of isotopic differences between the consumer and its diet. However, many empirical ecologists are still unfamiliar with the scope of applicability and the interpretative power of AA-SIR. To fill these knowledge gaps, we here describe a comprehensive approach to both carbon and nitrogen AA-SIR assessment focusing on two key topics: pattern in AA-isotope composition across spatial and temporal scales, and a certain variability of AA-specific isotope differences between the diet and the consumer. On this basis we review the versatile applicability of AA-SIR to improve our understanding of physiological processes as well as food web functioning, allowing us to reconstruct dominant basal dietary sources and trace their trophic transfers at the specimen and community levels. Given the insightful and opportunities of AA-SIR, we suggest future applications for the dual use of carbon and nitrogen AA-SIR to study more realistic food web structures and robust consumer niches, which are often very difficult to explain in nature.

Stepwise Approach for Tracing the Geographical Origins of the Manila Clam Ruditapes philippinarum Using Dual-Element Isotopes and Carbon Isotopes of Fatty Acids.

While there are many studies that have reported methods for tracing the geographical origin of seafoods, most of them have focused on identifying parameters that can be used effectively and not the direct application of these methods. In this study, we attempted to differentiate the geographical origins of the Manila clam R. philippinarum collected from different sites in Korea, the Democratic People's Republic of Korea, and China using a combination of analyses based on dual-element isotopes, fatty acids (FAs), and compound-specific isotopic analysis of FAs. We hypothesized that a stepwise application of new parameters to unclassified samples could achieve this objective by integrating new information while reducing time and labor. The FA profiles and compound-specific carbon isotopic values of FAs were found to enhance the discrimination power of determining the geographic origin up to 100%. Our findings demonstrate the advantageousness of using several parameters simultaneously over the conventional method of employing individual analytical methods when identifying geographic origins of the Manila clam, which could have implications for tracing the origins of different shellfish species or other food products as well.

Environmental fate and trophic transfer of synthetic musk compounds and siloxanes in Geum River, Korea: Compound-specific nitrogen isotope analysis of amino acids for accurate trophic position estimation.

Despite the extensive usage of synthetic musk compounds (SMCs) and siloxanes in various personal care products (PCPs), trophic magnification of such chemicals in aquatic environments remains unexplored. In June and September 2020, eleven SMCs and nineteen siloxanes were measured in water, sediments, and biota. Samples were collected from two sites where levels were expected to be influenced by the distance from the wastewater treatment plant (WWTP) in the Geum River, Republic of Korea, were expected. High concentrations of SMCs and siloxanes entered through WWTP were measured in water, sediment, and biota at the both sites and both seasons. The δ15N of amino acids provided a high-resolution food web and accurate trophic position (TP), which is an important factor for determining the trophic magnification factor (TMF). Among 24 TMFs, 19 of them were <1, ranging 0.7-0.8 for 1,3,4,6,7,8­hexahydro­4,6,6,7,8,8­hexamethyl­cyclopenta­Î³­2­benzopyran (HHCB), 0.6-0.8 for 6-Acetyl-1,1,2,4,4,7-hexamethyltetralin (AHTN), 0.7-0.8 for 4-tert-Butyl-3,5-dinitro-2,6-dimethylacetophenone (MK), 0.7-0.9 for octamethylcyclotetrasiloxane (D4), 0.1-0.4 for decamethylcyclopentasiloxane (D5), and 0.04-0.8 for dodecamethylcyclohexasiloxane (D6), and the remaining ones including HHCB, AHTN, MK, and D4 showed values close to 1 or slightly higher (TMF range: 1.0-2.3) indicating no or a little trophic magnification. The TMFs of these compounds were constant across sites and seasons. The TMF values of PCPs might be affected by species specificity and food web structure rather than by chemical properties such as log Kow, which describes a wide range of TMF values in various environments. This study presents valuable implications for assessing risk and managing environmental fate and trophic transfer of SMCs and siloxanes in freshwater environments.

Molecular evidence for suppression of swimming behavior and reproduction in the estuarine rotifer Brachionus koreanus in response to COVID-19 disinfectants.

The increased use of disinfectants due to the spread of the novel coronavirus infection (e.g. COVID-19) has caused burden in the environment but knowledge on its ecotoxicological impact on the estuary environment is limited. Here we report in vivo and molecular endpoints that we used to assess the effects of chloroxylenol (PCMX) and benzalkonium chloride (BAC), which are ingredients in liquid handwash, dish soap products, and sanitizers used by consumers and healthcare workers on the estuarine rotifer Brachionus koreanus. PCMX and BAC significantly affected the life table parameters of B. koreanus. These chemicals modulated the activities of antioxidant enzymes such as superoxide dismutase and catalase and increased reactive oxygen species even at low concentrations. Also, PCMX and BAC caused alterations in the swimming speed and rotation rate of B. koreanus. Furthermore, an RNA-seq-based ingenuity pathway analysis showed that PCMX affected several signaling pathways, allowing us to predict that a low concentration of PCMX will have deleterious effects on B. koreanus. The neurotoxic and mitochondrial dysfunction event scenario induced by PCMX reflects the underlying molecular mechanisms by which PCMX produces outcomes deleterious to aquatic organisms.

Validation of reference genes for quantitative real-time PCR in chemical exposed and at different age's brackish water flea Diaphanosoma celebensis.

Real-time quantitative reverse transcription polymerase chain reaction (qRT-PCR), a primary approach for evaluating gene expression, requires an appropriate normalization strategy to confirm relative gene expression levels by comparison, and rule out variations that might occur in analytical procedures. The best option is to use a reference gene whose expression level is stable across various experimental conditions to compare the mRNA levels of a target gene. However, there is limited information on how the reference gene is differentially expressed at different ages (growth) in small invertebrates with notable changes such as molting. In this study, expression profiles of nine candidate reference genes from the brackish water flea, Diaphanosoma celebensis, were evaluated under diverse exposure to toxicants and according to growth. As a result, four different algorithms showed similar stabilities of genes for chemical exposures in the case of limited conditions using the same developmental stage (H2A was stable, whereas Act was fairly unstable in adults), while the results according to age showed a significantly different pattern in suite of candidate reference genes. This affected the results of genes EcRA and GST, which are involved in development and detoxification mechanisms, respectively. Our finding is the first step towards establishing a standardized real-time qRT-PCR analysis of this environmentally important invertebrate that has potential for aquatic ecotoxicology, particularly in estuarine environments.

Application of Compound-Specific Isotope Analysis in Environmental Forensic and Strategic Management Avenue for Pesticide Residues.

Unintended pesticide pollution in soil, crops, and adjacent environments has caused several issues for both pesticide users and consumers. For users, pesticides utilized should provide higher yield and lower persistence while considering both the environment and agricultural products. Most people are concerned that agricultural products expose humans to pesticides accumulating in vegetation. Thus, many countries have guidelines for assessing and managing pesticide pollution, for farming in diverse environments, as all life forms in soil are untargeted to these pesticides. The stable isotope approach has been a useful technique to find the source of organic matter in studies relating to aquatic ecology and environmental sciences since the 1980s. In this study, we discuss commonly used analytical methods using liquid and gas chromatography coupled with isotopic ratio mass spectrometry, as well as the advanced compound-specific isotope analysis (CSIA). CSIA applications are discussed for tracing organic pollutants and understanding chemical reactions (mechanisms) in natural environments. It shows great applicability for the issues on unintended pesticide pollution in several environments with the progress history of isotope application in agricultural and environmental studies. We also suggest future study directions based on the forensic applications of stable isotope analysis to trace pesticides in the environment and crops.

Integrated approach for quantitative estimation of particulate organic carbon sources in a complex river system.

Despite receiving a considerable amount of attention in the past, quantitative and systematic estimation of the source contributions for different organic carbons (OCs) in complex river systems is still challenging. In this study, we tested an integrated framework using field data of bulk elements and lipid biomarkers and hydrological modeling (hydrological simulation program FORTRAN, HSPF) for the quantitative estimation of OC loads along different land-use types of a watershed (Geumho River watershed in South Korea). Based on the specific source assignments identified from the lipid biomarker patterns in particulate organic carbon (POC) such as short/long chains of alkanes, fatty acids and alcohols, and coprostanol/cholesterol, spatial variations of the diagnostic lipids could be used as an indicator to discriminate between the contributions of natural (algae, bacteria, and terrestrial plants) and anthropogenic sources (fecal). Based on the integration of HSPF modeling, it was also found that various POC loads might be partially controlled by different water discharges within watersheds. With the increase in POC fluxes, the increase in fecal loads was also noticed, as reflected by the predominant lipid (especially coprostanol normalized by water discharges). As a straightforward approach, we developed a set of indices including fecal index-1, ratios of coprostanol, fatty acids, and alkanes, which strengthened the sensitivity for fecal contamination. Compared with the conventional HSPF results, the variations of these proposed indices were more influenced by the broadened watershed extents with increasing downstream distance, which provided a more accurate estimation of the quantitative contributions of POC loadings in the complex river system.

A Multi-Elements Isotope Approach to Assess the Geographic Provenance of Manila Clams (Ruditapes philippinarum) via Recombining Appropriate Elements.

The increasing global consumption of seafood has led to increased trade among nations, accompanied by mislabeling and fraudulent practices that have rendered authentication crucial. The multi-isotope ratio analysis is considered as applicable tool for evaluating geographical authentications but requires information and experience to select target elements such as isotopes, through a distinction method based on differences in habitat and physiology due to origin. The present study examined recombination conditions of multi-elements that facilitated geographically distinct classifications of the clams to sort out appropriate elements. Briefly, linear discriminant analysis (LDA) analysis was performed according to several combinations of five stable isotopes (carbon (δ13C), nitrogen (δ15N), oxygen (δ18O), hydrogen (δD), and sulfur (δ34S)) and two radiogenic elements (strontium (87Sr/86Sr) and neodymium (143Nd/144Nd)), and the geographical classification results of the Manila clam Ruditapes philippinarum from Democratic People's Republic of Korea (DPR Korea), Korea and China were compared. In conclusion, linear discriminant analysis (LDA) with at least four elements (C, N, O, and S) including S revealed a remarkable cluster distribution of the clams. These findings expanded the application of systematic multi-elements analyses, including stable and radiogenic isotopes, to trace the origins of R. philippinarum collected from the Korea, China, and DPR Korea.

Combined effects of heavy metals (Cd, As, and Pb): Comparative study using conceptual models and the antioxidant responses in the brackish water flea.

The combined effect of toxic inducers has emerged as a challenging topic, particularly due to their inconsistent impacts on the environment. Using toxic unit (TU) based on LC50 value, we investigated the 48 h acute toxicities of the following combinations: Cd + As, Cd + Pb, As + Pb, and Cd + As + Pb, and binary and ternary combined effects were interpreted using concentration addition (CA) and independent action (IA) model. The molecular effects of these combinations were further examined on the basis of gene expression (four GST and two SOD isoforms) and antioxidant enzymes activity (SOD and GST). The CA-predicted LC50 was similar to the observed results, indicating that the CA model is more applicable for evaluating the combined effects of the metal mixtures. Synergistic effects (ΣTULC50 < 0.8) were observed for the mixtures As + Pb and Cd + Pb, while additive effects (0.8 < ΣTULC50 < 1.2) were observed for the mixtures Cd + As + Pb and Cd + As. No antagonistic effects were observed in this study. Molecular biomarkers for oxidative stress caused by metals, as well as traditional endpoints such as lethality, have shown a clear response in assessing the toxicity of binary and ternary mixtures. This study opens up a new avenue for the use of biomarkers to assess the combined effects of metals in aquatic environments.

Oxidative stress responses in brackish water flea exposed to microcystin-LR and algal bloom waters from Nakdong River, Republic of Korea.

Microcystis blooms and the impact of their toxins, particularly microcystin (MC), in coastal ecosystems is an emerging threat, but the species-specific effects of MC and the potential for bioconcentration are not fully understood. We exposed the brackish water flea, Diaphanosoma celebensis, to MC-LR, which showed antioxidant responses measured at the molecular to enzyme levels but no acute toxicity. We extended our experimental investigation to measure the released MC and its uptake by D. celebensis exposed to river water. In a short-term exposure (48 h) experiment, D. celebensis exposed to water from an algal bloom (approximately 2 µg L-1 MC) assimilated more than 50 pg MC per individual. The significant increase of MCs suggests the potential for the species to accumulate MCs. The dose-dependent increase in the antioxidant response observed in the mRNA levels also showed that D. celebensis exposed to diluted algal bloom waters were affected by toxins from cyanobacteria.

Trophic transfer of persistent toxic substances through a coastal food web in Ulsan Bay, South Korea: Application of compound-specific isotope analysis of nitrogen in amino acids.

Trophic magnification factor (TMF) of persistent toxic substances (PTSs: Hg, PCBs, PAHs, and styrene oligomers (SOs)) in a coastal food web (12 fish and four invertebrates) was determined in Ulsan Bay, South Korea. The nitrogen stable isotope ratios (δ15N) of amino acids [δ15NGlu-Phe based on glutamic acid (δ15NGlu) and phenylalanine (δ15NPhe)] were used to estimate the trophic position (TPGlu-Phe) of organisms. The TPGlu-Phe of organisms ranged from 1.64 to 3.69, which was lower than TP estimated by δ15N of bulk particulate organic matter (TPBulk: 2.46-4.21). Mercury and CB 138, 153, 187, and 180 were biomagnified through the whole food web (TMF > 1), while other PTSs, such as PAHs and SOs were not (biodilution of SOs firstly reported). In particular, the trophic transfer of PTSs was pronounced in the resident fish (e.g., rock bream, sea perch, Korean rockfish). Of note, CB 99, 101, 118, and 183 were additionally found to be biomagnifying PTSs in these species. Thus, fish residency appears to represent an important factor in determining the TMF of PTSs in the coastal environment. Overall, δ15NGlu-Phe provided accurate TPs of organisms and could be applied to determine the trophic transfer of PTSs in coastal food webs.

Cloning and molecular characterization of estrogen-related receptor (ERR) and vitellogenin genes in the brackish water flea Diaphanosoma celebensis exposed to bisphenol A and its structural analogues.

Although it has previously been shown that bisphenol (BP) analogues may interfere with the normal hormonal regulation by acting as endocrine disrupting chemicals (EDCs), little information is available on effects of BP analogues in invertebrates, particularly on cladocerans. In the present study, we identified estrogen-related receptors (EER), vitellogenin (VTG), and VTG receptor (VtgR) from the brackish water flea Diaphanosoma celebensis, and examined the effects of BPA and the substitutes, BPF and BPS, in different sublethal concentrations. Gene expression varied with time well matched with brooding, suggesting that DcEER, DcVTG, and DcVtgR play a role in reproduction in D. celebensis. qRT-PCR analysis showed that BPA and its substitutes differently modulated mRNA expressions of DcEER, DcVTG, and DcVtgR, indicating that these compounds adversely affect the normal reproduction-related pathway. This study facilitates better understanding of the molecular mode of action of BP analogues on the reproductive system of D. celebensis.

Variability of trophic magnification factors as an effect of estimated trophic position: Application of compound-specific nitrogen isotope analysis of amino acids.

The trophic magnification of persistent organic pollutants (POPs), which is the relationship between POP concentration and the trophic position (TPs) of an organism, is considered an important factor for prioritizing chemicals of concern in the environment. Organismal TPs are typically based on nitrogen isotope ratios of bulk tissue (δ15Nbulk). In this study, nitrogen isotope ratios of amino acids (δ15NAAs), a more precise approach for TP estimation (TPAAs), was applied and compared with estimations of TP based on δ15Nbulk (TPbulk) in marine organisms living in Masan Bay, South Korea. Compound-specific isotope analysis of the amino acids (CSIA-AAs) in fish samples allows us to calculate robust TPs by correcting the variation in baseline isotope values with use of the δ15Nbulk technique. In a benthic food chain, this approach reveals more significant magnification trends for POPs [polychlorinated biphenyls (PCBs), polybrominated diphenyl ethers (PBDEs), and organochlorine pesticides (OCPs)] than those revealed by analysis of the relationship between TPbulk and POPs. The trophic magnification factors (TMF) associated with TPAAs were significant for some POPs, especially pp'-DDD and chlordane. The results presented in this study suggest that TP calculations based on δ15NAAs are an effective tool for characterizing trophic magnification trends related to the fates of various pollutants.

Carbon and nitrogen stable isotope signatures linked to anthropogenic toxic substances pollution in a highly industrialized area of South Korea.

Sources of organic matter in a polluted coastal embayment (Lake Sihwa) indicated that δ13C and δ15N of suspended particulate matter (SPM) and sediments could be distinguished by land-use type and surrounding activities. Specifically, low δ15N occurred in inland creeks near industrial complex, where severe contamination by persistent toxic substances (PTSs) is evidenced. To identify the sources of 15N-depleted organic matter and PTSs, the SPM and/or sediments were collected along ~8 km of the Singil Creek and in stormwater drainage pipes that carried pollutants to the creeks from upland areas. Results indicated that stormwater originating in industrial areas was the main source of low δ15N and elevated PTSs, which appeared to flow into stormwater drains with rainwater and eventually into the creeks at stormwater outfalls. To improve the water quality of inland creeks, it will be necessary to reduce the amount of untreated stormwater entering stormwater drainage system from industrial complex.

Environmental assessment of contaminated marine sediments treated with solidification agents: Directions for improving environmental assessment guidelines.

Treatment of dredged materials is a critical issue, since management and disposal of these products requires considerable investment of monetary resources, time, and space. The high concentration of pollutants in dredged materials, along with high water content and many fine particles make recycling these materials particularly difficult. In order to solve this problem, solidification/stabilization has been considered as a potentially viable solution for recycling dredged marine sediments. However, there are currently no guidelines that address potential biological and environmental impacts. To evaluate the stability of treated materials and their biological impacts, dredged marine sediments, which were polluted with heavy metals, were treated by solidification/stabilization using two different solidifying agents. To assess potential impacts, toxicity characteristic leaching procedures (TCLP, USEPA) and a bioassay (with the rotifer, Brachionus sp.) were performed with treated materials. In a TCLP test, we found that treatment with a solidification agent decreased the leaching concentration of heavy metals from sediment compared to the control. The rotifer bioassay showed no change in the survival rate during 24 h of exposure to both agents. However, survival differed between the two agents after 48 h of exposure. Screening physiological status using gene expression, showed that oxidative stress genes were significantly altered. These results suggest that more studies are needed to provide guidelines for deciding the usability of treated materials created by the solidification or stabilization of dredged materials.