Exposure to and Biomarker Responses From Legacy and Emerging Contaminants Along Three Drainages in the Milwaukee Estuary, Wisconsin, USA.

Legacy contaminants and contaminants of emerging concern (CECs) were assessed in tree swallow (Tachycineta bicolor) tissue and diet samples from three drainages in the Milwaukee estuary, Wisconsin, USA, to understand exposures and possible biomarker responses. Two remote Wisconsin lakes were assessed for comparative purposes. Bioaccumulative classes of contaminants, such as polybrominated diphenyl ethers and per- and polyfluoroalkyl substances, while at higher concentrations than the reference lakes, did not vary significantly among sites or among the three drainages. Polycyclic aromatic hydrocarbons were assessed in diet and sediment and were from primarily pyrogenic sources. Ten biomarkers were assessed relative to contaminant exposure. Polychlorinated biphenyls (PCBs) were elevated above reference conditions at all Milwaukee sites but did not correlate with any measured biomarker responses. Only one site, Cedarburg, just downstream from a Superfund site, had elevated PCBs compared to other sites in the Milwaukee estuary. Few non-organochlorine insecticides or herbicides were detected in tree swallow liver tissue, except for the atrazine metabolite desethylatrazine. Few pharmaceuticals and personal care products were detected in liver tissue except for N,N-diethyl-meta-toluamide, iopamidol, and two antibiotics. The present study is one of the most comprehensive assessments to date, along with the previously published Maumee River data, on the exposure and effects of a wide variety of CECs in birds. Environ Toxicol Chem 2024;43:856-877. © 2024 SETAC. This article has been contributed to by U.S. Government employees and their work is in the public domain in the USA.

Concentration-dependent effects of lithium on Daphnia magna: Life-history profiles and integrated biomarker response implementation.

The growing use of lithium (Li) in industrial and energy applications and increasing demand worldwide has inevitably resulted in its wide dispersal, representing a significant threat to aquatic systems. Unfortunately, as a ubiquitous emerging contaminant, the comprehensive toxicological information regarding Li at multifarious levels is limited. To diminish this gap, this work was focused to explore Li-induced cascading effects on Daphnia magna as a key species in freshwater ecosystems. Specifically, the organisms were chronically exposed to gradient Li concentrations with emphasis on characterizing life-history traits from individual to population scale, primarily as observed by a markedly concentration-dependent decrease along exposure gradients. In parallel, a robust set of biomarkers relating to energy reserves, antioxidant and biotransformation enzymes, cellular damage, ionoregulation and neurotoxicity were assayed for further understanding potential underlying mechanisms. As a result, biomarker alterations were characterized by significant decreases in energy storage and enzymatic profiles of antioxidant and biotransformation systems, not only triggering an imbalance between reactive oxygen species (ROS) generation and elimination under Li exposure, but compromising the fecundity fitness of phenotypical costs. In contrast, malondialdehyde (MDA) levels were remarkably enhanced as a consequence of inefficient antioxidant and biotransformation capacity leading to lipid peroxidation (LPO). Additionally, Li exerted a dose-dependent biphasic effect on the activities of superoxide dismutase (SOD), Na+,K+-ATPase and acetylcholinesterase (AChE) by interfering with inherent balance. In terms of responsive patterns and dose-effect trends, the integrated biomarker response indices (IBRv2) and star plots were consistent with the differences in biomarker profiles, not only presenting comprehensively biological effects in a visualized form, but signaling the importance of progressive induced changes in an integrative way. Overall, these findings highlighted the need for elucidating Li-produced impacts from a comprehensive perspective, providing valuable insights into better understanding the toxicity of Li in relation to aquatic ecosystem functioning and ecological relevance.

Integrated biomarker responses: a further improvement of IBR and IBRv2 indexes to preserve data variability in statistical analyses.

In biomonitoring and laboratory studies, it is typical to measure a battery of molecular, biochemical, and cytogenetic biomarkers to evaluate the effects of xenobiotics in biota. However, summarizing the results of several biomarkers to inform laypersons and environmental agencies is still a challenge for researchers and environmental specialists. To address this issue, researchers have developed indexes such as the Integrated Biomarker Responses (IBR) and Integrated Biomarker Response version 2 (IBRv2) to summarize all biomarkers responses into a single value. Unfortunately, these indexes do not preserve the original biological variability, which hampers subsequent statistical analyses. In this study, we created new versions of IBR and IBRv2, which preserve individual data variability and can be used in typical statistical analyses. The new Integrated Biomarker Responses individual (IBRi), Integrated Biomarker Responses version 2 individual (IBRv2i) and Weighted Integrated Biomarker Responses version 2 individual (Weighted IBRv2i) indexes correlated with the original IBR and IBRv2 indexes and were able to detect differences among experimental groups in a simulated and case studies. Using the IBRi, IBRv2i, or Weighted IBRv2i indexes is advantageous because they maintain the data variability of the experimental groups and can be analyzed using hypothesis testing statistics like any other parameter. Additionally, this approach can help translate technical scientific terminology into a more accessible language suitable for environmental governmental agencies and decision-makers.

Responses and detoxification mechanisms of earthworm Amynthas hupeiensis to metal contaminated soils of North China.

Metal contamination is widespread, but only a few studies have evaluated the toxicological risks of metals (Cd, Cu, and Pb) in earthworms from farmlands in North China (Hebei province). Amynthas hupeiensis, the dominant species in the study area, was used to determine the responses and detoxification mechanisms of uncontaminated (CK), and low (LM)-, and high (HM)-metal-contaminated soils following 7-, 14-, and 28-days exposure. Metal toxicity in LM and HM soils inhibited the biomass of A. hupeiensis. The concentrations of Cd in A. hupeiensis bodies indicated accumulated Cd appeared to remain steady with prolonged exposure, while Cu/Pb increased significantly with soil levels. Bioaccumulation occurred in the order Cd > Pb > Cu in LM soil, and in the order Cd > Cu ≈ Pb in HM soil, which was attributed to differences in available fractions between LM and HM soils. Physiological levels of biomarkers in A. hupeiensis were determined, including total protein (TP), glutathione (GSH), glutathione peroxidase (GPx), acetylcholinesterase (AChE), and malondialdehyde (MDA). Deviations in GSH, GPx, and AChE were considered to denote sensitive biomarkers using the IBRv2 index. Metabolomics data (1H nuclear magnetic resonance-based) revealed changes in metabolites following 28-days exposure to LM and HM soils. Differences in metabolism in A. hupeiensis following exposure to LM and HM were related to energy metabolism, amino acid biosynthesis, glycerophospholipid metabolism, inositol phosphate metabolism, and glutathione metabolism. Metal stress from LM and HM soils disturbed osmoregulation, resulting in oxidative stress, destruction of cell membranes and inflammation, and altered levels of amino acids required for energy by A. hupeiensis. These findings provide biochemical insights into the physiological and metabolic mechanisms underlying the ability of A. hupeiensis to resist metal stress, and for assessing the environmental risks of metal-contaminated soils in farmland in North China.

Toxicity analysis and biomarker response of Quinalphos Organophosphate Insecticide (QOI) on eco-friendly exotic Eudrilus eugeniae earthworm.

An ever-increasing use of pesticides in agricultural fields has led to a catastrophic decline in crop quality and, ultimately soil fertility. To control various pests, quinalphos is commonly used in India's tea plantations. This study aims to investigate the effects of the Quinalphos organophosphate insecticide on the non-target beneficial organism Eudrilus eugeniae earthworms and the biomarkers that respond to its effects. Earthworm species, especially E. eugeniae, remains as the most trustworthy and well-suited model organism for conducting a wide variety of environmental studies. The median lethal concentration (LC50) was identified as 3.561 µg cm-2 (contact filter paper) and 1.054 mg kg-2 (artificial soil toxicity). The 5% and 10% of LC50 value 3.561 µg cm-2 was exposed to earthworm to analyze the sublethal effects at pre-clitellum, clitellum, and post-clitellum segments. Specific enzymatic activities of neurotransmitter enzyme acetylcholinesterase; antioxidant enzymes such as lipid peroxidase, superoxide dismutase, and catalase; and detoxification enzymes including glutathione S transferase, reduced glutathione, carboxylesterase, and Cytochrome P450 were analyzed. Exposure of E. eugeniae earthworm to subacute exposures of pesticides caused significant alterations in these stress markers in a concentration-dependent manner. Morphological abnormalities like bulginess, coiling, and bleeding were observed after exposure of the insecticide treatments. Histological cellular disintegration, a reduced NRRT time, and an inhibited proteolytic zone were also identified in pesticide-exposed earthworms. Studies demonstrate that the organophosphate insecticide quinalphos causes acute toxicity in E. eugeniae; hence, it is suggested that non-target eco-friendly E. eugeniae earthworms may be at risk if exposed to the excessive concentrations of quinalphos organophosphate insecticide in soil.

New insights into the effects of chlorpyrifos on soil microbes: Carbon and nitrogen cycle related microbes in wheat/maize rotation agricultural field.

Chlorpyrifos, a broad-spectrum organophosphorus insecticide, has been widely detected worldwide and is a potential neurotoxin and endocrine disruptor. Besides, chlorpyrifos has been proven that have a negative effect on soil microbes. In the present study, chlorpyrifos formulation (LORSBAN®, 45% emulsifiable concentrate) was applied in an agricultural field at the recommended dose (R dose, 270.0 and 337.5 g a.i. ha-1 for wheat and maize respectively) and double recommended (DR) dose. Chlorpyrifos residue level and effect on soil microbes related to soil carbon and nitrogen cycle function were analyzed. Results showed that the half-lives of chlorpyrifos in wheat and maize field soil were 7.23-8.23 and 1.45-1.77 d, respectively. Application of chlorpyrifos at even DR dose did not result in unacceptable residual chlorpyrifos, where the final residual chlorpyrifos in wheat/maize (leaf, stem, and grain) was meet the requirement of the maximum residual limit (0.5 mg kg-1 for wheat and 0.05 mg kg-1 for maize) in China. Chlorpyrifos enhanced the activity of ß-glucosidase by increasing the relative abundance of Sphingosinicella and promoted the carbon cycle in wheat field. The changes of cbbLR and cbbLG gene abundance also confirmed that chlorpyrifos could affect the import and export of soil carbon pool. The effect of chlorpyrifos on soil N cycle was determined by changes in the abundance of the bacterial genus Gemmatimonas, which is associated with denitrification. Further analysis of N-cycle functional genes and urease activity showed that chlorpyrifos inhibited nitrogen fixation in wheat field, but promoted nitrogen fixation in maize field. In general, bacterial abundance, urease, and AOA-amoA gene could be early warning markers of chlorpyrifos contamination. The results demonstrated the negative effects of chlorpyrifos on soil microbes especially on soil C and N cycle in actual agricultural field. It provides new insights about chlorpyrifos environmental pollution and its effect on soil ecosystems.

Spatial distribution of benthic toxicity and sediment-bound metals and arsenic in Guangzhou urban waterways: Influence of land use.

The effects of land use on pollutant loads in sediments have been well documented; however, its influence on spatial variations in sediment toxicity remains largely unknown. In the present study, the toxicological effects of 17 sediments collected from Guangzhou waterways were evaluated using two benthic invertebrates (Chironomus dilutus and Hyalella azteca), along with quantification of heavy metals and arsenic in the sediments. The impacts of land-use configuration on sediment toxicity and occurrence of heavy metals and arsenic were analyzed. The sediments presented moderate lethality (<40 %) in the two test species and significantly altered their enzymatic activity, including the activities of oxidative stress biomarkers and acetylcholine esterase. Metals (Ag, Cd, Cr, Cu, Hg, Ni, Pb, and Zn) and arsenic were detected in all the sediments, with total concentrations ranging from 238 to 1019 mg/kg of dry weight. Both the toxicity and chemical results displayed spatially dependent patterns but were related to different land use types. Toxicity was most influenced by agricultural and aquacultural activities, while metal and arsenic pollution was most influenced by urban land areas. The present findings are expected to provide essential knowledge for developing strategies that reduce the chemical pollution and ecotoxicological risk in sediments.

An integrated biomarker assessment of biochemical responses in a freshwater fish species after vanadium pentoxide (V2O5) exposure.

Vanadium (V) is a toxicant becoming increasingly concentrated in freshwater with the potential to affect aquatic organisms. Vanadium pentoxide (V2O5), accumulated in fish, can act as an oxidizing agent and cause oxidative damage. To determine the effects of V2O5 on exposed adult Oreochromis mossambicus, acute exposure experiments were conducted. Bioaccumulation and biomarker analyses were performed on various excised tissues of the exposed fish. As expected, accumulated V concentrations in the gills increased as the exposure concentration increased. Gill tissue accumulated more vanadium than muscle tissue. Metallothionein content increased in the highest concentrations compared to the lower concentrations, therefore showing that metallothionein proteins were attempting to sequester V2O5 in the tissues. Superoxide dismutase (SOD) showed an excitation at lower concentrations and inhibition as the exposure concentrations increased, possibly due to ROS detoxification. Catalase activity decreased from the first exposure concentration to the last concentration; this could have been due to SOD compensation. Protein carbonyl concentration decreased as the concentrations of V2O5 increased, indicating an inhibition of protein oxidation. The IBRv2 comparison revealed the biochemical responses caused by V2O5 more effectively than traditional statistical analysis.

Assessment of the exposure of two pesticides on common carp (Cyprinus carpio Linnaeus, 1758): Are the prolonged biomarker responses adaptive or destructive?

Chlorpyrifos (CPF) and cypermethrin (CYP) are two insecticides that have a proven negative effect on non-target aquatic organisms when they enter the surface waters. However, literature on the comparative effects of these pesticides on important aquaculture fish species, such as common carp (Cyprinus carpio Linnaeus, 1758) is not yet scientifically detailed, especially over the long-term. The idea of conducting a long-term exposure is to find out how the observed biomarkers would change compared to the short-term exposure. In the natural environment, toxicants are not present alone, but in combination. By monitoring the long-term impact of individual substances, the state of aquatic ecosystems exposed to various toxicants could be predicted. Thus, this study aimed to evaluate the toxicity of different concentrations of CYP (0.0002, 0.0003, and 0.0006 µg/L) and CPF (0.03, 0.05, and 0.10 µg/L) in 50-L glass tanks on C. carpio, exposed for 30 days under laboratory conditions. A set of histological and biochemical biomarkers in the gills and liver were applied with the chemical analyses of water and fish organs. Furthermore, the condition and hepatosomatic index were calculated to assess the physiological status of the treated carps. The behavioral responses were also monitored, and the respiration rate was analyzed. The results suggest that CYP had a more prominent effect on the histological structure of fish organs, biochemical responses of anti-oxidant enzymes, behavior, and respiration rate compared to the effect of CPF. In addition, the results also indicate that the liver is more susceptible to chronic and chemically induced cellular stress compared to the gills, with overall destructive changes in the histological biomarkers rather than adaptive. Regardless of the scenario, our results provide novel insights into pesticide exposure and the possible biological impacts on economically important freshwater fish, exposed to lower CYP and CPF concentrations, based on the EU legislation (maximum allowable concentrations, MAC-EQS).

Metabolism toxicity and susceptibility of decabromodiphenyl ether (BDE-209) exposure on BRL cells with insulin resistance.

Type 2 diabetes mellitus (T2DM) is a metabolic disease characterized by insulin resistance (IR) and has attracted worldwide attention due to its high prevalence. As a typical persistent organic pollutant, decabromodiphenyl ether (BDE-209) has been detected in food and human samples, and the concentration trends increase year by year. In addition, it has been proved to have the potential to increase the risk of IR, but it is rarely reported whether it could aggravate IR in T2DM. Therefore, in this study, the IR-BRL (buffalo rat liver cells with IR) model was applied to study the metabolism toxicity and susceptibility of BDE-209. Results showed that BDE-209 could inhibit glucose absorption and increase the levels of serum total cholesterol (TC) and triglyceride (TG), ultimately leading to the disorder of glucolipid metabolism in IR-BRL cells. Besides, it also could cause cell damage by increasing the levels of aspartate transaminase (AST), alanine aminotransferase (ALT), and malondialdehyde (MDA) in cells. Moreover, its potential mechanisms were to: (1) affect the transport of glucose, synthesis of glycogen and fatty acid via IRS-1/GLUT4 and IRS-1/PI3K/AKT/GSK-3ß pathways; (2) impact the proliferation and differentiation by regulating the expression of Mek1/2, Erk1/2, and mTOR proteins and genes. Furthermore, susceptibility analysis showed that there was a significant synergism interaction between IR and BDE-209, which suggested that IR-BRL cells were more susceptible to the metabolism toxicity induced by BDE-209.

Integration assay of bacteriological risks in marine environment using salmonella spp and multimarker response in the bivalve Donax trunculus: Novel biomonitoring approach.

Microbiological contamination is one of the riskiest forms of human contamination in seawater, which threaten the stability of ecosystems and human health. In this study, we study the accumulation of a pathogenic bacteria Salmonella spp; isolated from the marine environment, in the soft tissue of Donax trunculus (Mollusca, Bivalvia), a commonly used as a bioindicators species for aquatic ecosystems monitoring, under laboratory conditions during both exposure and recovery periods. These bacteria were added in seawater at three concentrations previously determined against sentinel specie at three exposure periods (24, 48 and 96 h). In a second series of experiments, exposed specimens were afterward transplanted to clean water to assess the recovery pattern. The mortality rate of bivalves was determined as biomarker of general stress. Our findings suggest that microbiological contamination by Salmonella spp was gradually incorporated into the body of D. trunculus causing a significant induction of enzymatic activity of acetylcholinesterase (AChE), Catalase (CAT), glutathione-S-transferase (GST) and malondialdehyde (MDA) levels, as a function of time and concentration. Exposure to a bacterial concentration of 5.104 bacteria/liter resulted in the mortality of more than 80% of the specimens. This study is to test the pathogenicity of Salmonella strains at concentrations close to those of the marine environment, and their effects on biomarkers, thus deducing the existence of an exponential relationship between bacterial concentrations and enzymatic response. The principal component analysis shows that the four biomarkers had similar variation with bacterial concentrations, while two groups were obtained to change following the exposure time (CAT-GST and AChE-MDA). This study provides new findings on the potential accumulation of pathogenic bacteria associated with neurotoxicity and oxidative stress in the wedge clam Donax trunculus.

Effects of short-term exposure to tralopyril on physiological indexes and endocrine function in turbot (Scophthalmus maximus).

Tralopyril is an emerging marine antifouling agent with potential toxic effects on non-target aquatic organisms. To evaluate the toxicity of tralopyril, to turbot (Scophthalmus maximus), we assessed biomarkers, including oxidative stress, neurotoxicity, and osmotic homeostasis regulation enzymes, after a 7-day exposure to tralopyril (5 µg/L, 15 µg/L, 30 µg/L). Superoxide dismutase activity was significantly decreased at 30 µg/L, and Ca2+-Mg2+-ATPase activity in the gills was significantly increased at 15 µg/L and 30 µg/L. No statistically significant differences in the responses of acetylcholinesterase and nitric oxide were detected. In addition, 15 µg/L and 30 µg/L tralopyril induced hyperthyroidism, reflected by significantly increased of T3 levels. The expression levels of hypothalamus-pituitary-thyroid axis-related genes were also upregulated. The molecular docking results showed that the thyroid system disruption was not caused by competitive binding to the receptor. In addition, the integrated biomarker response index showed that 15 µg/L tralopyril had the greatest effect on turbot. In general, tralopyril caused oxidative damage, affected energy metabolism, and interfered with the endocrine system. These findings could provide reference data for assessing the ecological risk of tralopyril in marine environments.

Short-term exposure of decabromodiphenyl ether in female adult Balb/c mice: Immune toxicity and self-recovery.

As a typical persistent organic pollutant, decabromodiphenyl ether (BDE-209) is associated with various health risks, especially on immune system, which is sensitive to environmental pollutants. In addition, there is a problem of multi-index estimation and lack of comprehensive evaluation in immune toxicity study. In this study, the immunotoxicity of BDE-209 was systematically estimated from the aspects of immunopathology, humoral immunity, cellular immunity and non-specific immunity, etc., and integrated biomarker responses (IBR) combined with principal component analysis was applied to comprehensively evaluate the immunotoxicity of BDE-209 and its self-recovery after discontinuation. Results showed that BDE-209 exposure could cause immunotoxicity. This response seems to depend on (1) atrophying immune organs (thymus and spleen), hepatomegaly accompanied by increasing aspartate aminotransferase and oxidative stress;(2) changing humoral (immunoglobulins) and cellular (lymphocyte proliferation and cytokine secretion) immunity indices; (3) altering related expressions of genes, and further leading to imbalance of Th1/Th2 (Th, helper T cell). Integrated biomarker responses (IBR) companied with principal component analysis selected five biomarkers (mRNA expression of GATA-3, malondialdehyde level in thymus, count of white blood cell, serum IgG and lipopolysaccharide-induced splenic lymphocyte proliferation) to clarify the immunotoxicity induced by BDE-209. Furthermore, IBR combined with factorial analysis revealed that the effect of BDE-209 could be dose-dependently reduced after withdrawal of BDE-209. Overall results suggested that BDE-209 has immunotoxicity on adult Balb/c mice, whereas this immunotoxicity could be reduced by the self-regulation of organisms to some extent.

Evaluating environmental impact of STP effluents on receiving water in Beijing by the joint use of chemical analysis and biomonitoring.

To evaluate the environmental impact of receiving water from the Qinghe River sewage treatment plant (STP) effluents in Beijing, we collected sediments and Bellamya aeruginosa (Up-site, Discharge-site, and Down-site) both in 2017 and 2018 and analyzed the samples via chemical analysis, biological responses and transcriptomics. In two years of data, our biological results showed that AChE activities presented different degrees of influence on B. aeruginosa captured at sampling points of the STP compared to control sites (P < 0.05). Additionally, indicators of the antioxidant system (e.g., SOD, CAT, GST, EROD activity) and MDA content were significantly increased in the whole tissue at the Up-site of the STP. Integration of the assessed biomarkers using the integrated biomarker response (IBR) index ranked the environmental impact at sites as Up-site > Discharge-site > Down-site. In terms of the transcriptome data, B. aeruginosa collected from the Discharge-site of the STP showed greater transcriptomic response than it did from all other sites. KEGG pathway analysis revealed that sewage significantly altered the expression of genes involved in xenobiotics by cytochrome P450, drug metabolism-cytochrome P450, glutathione metabolism, oxidative phosphorylation, citrate (TCA) cycle, glycolysis/gluconeogenesis, apoptotic and Parkinson's disease. The concentrations of 34 organic pollutants (17 PAHs, 10 PAEs, 7 EDCs) were measured. The chemical concentrations of pollutants decreased from Up-site to Down-site and were well correlated with enzyme activity, IBR, and transcriptomic results. Our results demonstrated that the combined use of chemical analysis, biological responses and transcriptome data is necessary to validate the efficacy of a battery of biomarkers chosen to detect environmental stress due to pollution.

Evaluation of joint toxicity of heavy metals and herbicide mixtures in soils to earthworms (Eisenia fetida).

It is widely acknowledged that a simplified and robust approach to evaluating thecombined effects of chemical mixtures is critical for ecological risk assessment (ERA) of contaminated soil. The earthworm (Eisenia fetida) was used as a model to study the combined effects of polymetallic contamination and the herbicide siduron in field soil using a microcosm experiment. The responses of multiple biomarkers, including the activities of catalase (CAT), superoxide dismutase (SOD), glutathione reductase (GR) and acetylcholine esterase (AChE), the concentrations of glycogen, soluble protein (SP), malonaldehyde (MDA), and metallothionein (MT), and the neutral red uptake test (NRU), were investigated. Multivariate analysis, Principal Component Analysis (PCA) and Spearman's Rank Correlations analysis (BVSTEP) revealed that the activities of AChE and CAT and the NRU content were the prognostic biomarkers capturing the minimum data set of all the variables. Internal Cd (tissue Cd) in earthworms was closely related to the health status of worms under combined contamination of heavy metals and siduron. The integrated effect (Emix) calculated based on the activities of AChE and CAT and NRU content using the stress index method had significantly linear regression with internal Cd (p<0.01). Emix(10), Emix(20), and Emix(50) were then calculated, at 1.27, 1.63 and 2.71 mg/kg dry weight, respectively. It could be concluded that a bioassay-based approach incorporating multivariate analysis and internal dose was pragmatic and applicable to evaluating combined effects of chemical mixtures in soils under the guidance of the top-down evaluation concept of combined toxicity.

Metallothionein, hemocyte status and superoxide dismutase/aspartate aminotransferase activity are sensitive biomarkers of cadmium stress in Onchidium reevesii.

Metal pollution in the environment is a serious threat to the biological sustainability of coastal ecosystems. However, our current understanding of the biological effects of metals in these ecosystems is limited. Herein, we investigated the responses of the sea slug Onchidium reevesii to persistent sublethal Cd environmental stress. Dynamic expression was analyzed using various biomarkers. The full-length cDNA of O. reevesii metallothionein (MT) was cloned and consists of 1639 nucleotides encoding a 65 amino acid polypeptide. Phylogenetic analysis showed that Or-MT has conserved Cys residues typical of MTs, including a typical Cys-X-Cys motif, implying that it can function the same as the MT of other shellfish. Expression of Or-MT in response to Cd varied in different tissues, and was highest in gastropod tissues. Thus, regiotemporal expression of MT may be useful for assessing pollution in coastal areas. Cellular immunity (in the hemolymph) and enzyme activity (in the hepatopancreas) were investigated along with hemocyte viability, hemocyte phagocytosis, and superoxide dismutase (SOD) and aspartate aminotransferase (AST) activities. Hemocyte viability was elevated under continuous Cd exposure but hemocyte phagocytosis was decreased. SOD and AST activities in the hepatopancreas fluctuated considerably, and SOD activity was more sensitive. SOD activity was lowest at 4 h and highest at 12 h, while AST activity peaked at 2 h and was lowest at 48 h. Thus, changes in enzyme activity may reveal adaptation to stress. Furthermore, the response patterns of certain enzymes, cellular immunity, and MT expression in O. reevesii could serve as biomarkers of Cd pollution in aquatic environments.

A multilevel response approach reveals the Asian clam Corbicula largillierti as a mirror of aquatic pollution.

The endobenthic bivalves are widely used as a bioindicators since they inhabit the sediment-water interface and are able to accumulate a different kind of contaminants. In the present work, we evaluated wild Corbicula largillierti (Phillippi, 1844) as a bioindicator of water quality in the central region of Argentina. The responses at different levels of the biological organization were used. We measured organochlorine pesticides (OCPs) and polychlorinated biphenyls (PCBs) levels in water and clams tissues. The biomarkers selected were enzymatic activities (Glutathione S-Transferase, Catalase, Acetyl-, Butyryl-cholinesterase, and Carboxylesterase) morphometry of the digestive gland, condition index and morphology of valves. In order to integrate all the responses a multivariate analysis and integrated stress index were applied. Our results showed the presence of contaminants along the studied river and the ability of C. largillierti to bioaccumulate them. All the biomarkers selected varied according to the water quality gradient, although there was no specific correlation with OCPs and PCBs levels. At the most polluted sites, the detoxification and oxidative stress enzymes, the morphometric analysis of the digestive gland and the variation in the morphology of the valves indicated the water quality degradation. The multivariate analyses allowed to discriminate the sites according to the different biomarker responses. The IBR index also showed a variation pattern according to the environmental quality gradient along the basin. According to the responses shown by C. largillierti we suggest this species as an useful bioindicator of aquatic pollution.

Biomarker responses in oysters Crassostrea hongkongensis in relation to metal contamination patterns in the Pearl River Estuary, southern China.

The Pearl River Estuary (PRE) is the third largest estuary in China, where estuarine organisms are under metal stress at various biological levels. Based on the metal concentrations measured in oyster Crassostrea hongkongensis, we documented a change in dominance of metal contamination from Cd, Cr, Cu, Ni and Zn to Ag, Cd, Cu and Zn. In general, metal concentrations were higher in upstream stations and displayed a clear up-downstream gradient. Compared to the historical values, we noted the reductions in Cd, Cr and Ni concentrations, and the changing inputs due to evolving industrial activities were responsible for shaping the metal contamination profile in the PRE region. Along with metal concentrations, a suite of biomarkers was analyzed. Among the metals measured in the oyster tissues, Ag, Cd, Cu, Ni and Zn showed the strongest associations with pro-oxidant and oxidative stress responses (superoxide dismutase, lipid peroxidation and lysosomal membrane destabilization) and detoxification responses (glutathione and metallothionein), suggesting that the present metal contamination still exerts significant amount of stress in biota in the PRE. Metal contamination in estuaries in China is still severe compared to other countries, therefore continuous efforts should be taken to monitor the changing metal profiles with necessary control and remediation measures.

Bioaccumulation and toxic effects of nanoparticulate and ionic silver in Saccostrea glomerata (rock oyster).

The increasing production of Ag nanoparticle (AgNP) containing products has inevitably led to a growing concern about their release into the aquatic environment, along with their potential behaviour, toxicity, and bioaccumulation in marine organisms exposed to NPs released from these products. Hence, this study is focused on the effects of AgNPs in Saccostrea glomerata (rock oyster) in artificial seawater (ASW); evaluating the NP's stability, dissolution, and bioaccumulation rate. AgNPs NM300K (20 ±â€¯5 nm) in concentrations of 12.5 µgL-1 and 125 µgL-1 were used to conduct the experiments, and were compared to a blank and a positive control of 12.5 µgL-1 AgNO3. Dissolution in ASW was measured by ICP-OES and stability was assessed by TEM after 1 h and 3, 5, and 7 days of exposure. Bioaccumulation in gills and digestive glands was measured after 7 days of exposure. The higher concentration of AgNPs induced more aggregation, underwent less dissolution, and showed less bioaccumulation, while the lower concentration showed less aggregation, more dissolution and higher bioaccumulation. Five biomarkers (EROD: ethoxyresorufin-o-deethylase, DNA strand breaks, LPO: lipid peroxidation, GST: glutathione S-transferase and GR: glutathione reductase) were analysed at 0, 3, 5 and 7 days. Significant differences compared to the initial day of exposure (day 0) were reported in DNA strand breaks after 5 and 7 days of exposure, GST, from the third day of exposure, in all the Ag samples, and in some samples for LPO and GR biomarkers, while no significant induction of EROD was observed. A combined effect for each type of treatment and time of exposure was also reported for DNA strand breaks and GST biomarkers measured at the digestive glands. In general, the significant inductions measured showed the following trend: 125 µgL-1 AgNPs >12.5 µgL-1 AgNPs ∼12.5 µgL-1 AgNO3 even though bioaccumulation followed the opposite trend.

Evaluation of combined toxicity of Siduron and cadmium on earthworm (Eisenia fetida) using Biomarker Response Index.

Agrochemicals and heavy metals are widespread contaminants in urban soil and could co-exist as mixture, which could cause unexpected risk to terrestrial organism. To assess the joint effect of herbicide Siduron and Cd, a battery of sub-lethal biomarkers was studied using earthworm ecotoxicological assay. Most selected biomarkers appeared significant but complicated responses with the increasing concentration of contaminants after 28-day exposure. In order to quantify the overall effect of the mixture contaminants, Biomarker Response Index (BRI) was used to integrate the multiple responses. Concentration Addition Index (CAI) and Effect Addition Index (EAI) were introduced to assess types of joint effect. Results showed significantly dose-effect responses between BRI and contaminant exposure concentrations. Integrated toxicity increased obviously under joint treatments of Siduron and Cd compared to their individual treatments. According to CAI, a clear antagonism was observed at relatively lower effects and gradually transformed to slight synergism with an increase of effects, while EAI showed the joint effect of addition at the whole range of effect levels. Thus, compared to the simple analysis of those complicated responses, BRI is an effective method to determine the integrated toxicity of mixture and its combination with joint effect indices (CAI and EAI) provides more worthy risk assessment on toxicity interaction among compounds.