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Microplastics are widespread pollutants in the environment and are considered a global pollution problem. Microplastics mostly originate from larger plastics and due to environmental conditions are undergoing constant fragmentation processes. It is important to understand the fragmentation pathways, since they play a key role in the fate of the particles, and also directly influence toxicity. Amphipods are potential inducers of plastic debris fragmentation. Here, Hyalella azteca was exposed to different concentrations (540, 2700, 5400 items/L) of 24.5 µm polystyrene microplastics (PS-MP) for 7 days. After exposure, oxidative stress, particle size reduction, and mortality were checked. No significant mortality was seen in any of the treatments, although changes were recorded in all enzymatic biomarkers analyzed. It was observed that throughout the ingestion and egestion of PS-MP by H. azteca, particles underwent intense fragmentation, presenting a final size up to 25.3% smaller than the initial size. The fragmentation over time (24, 72, 120, 168 h) was verified and the results showed a constant reduction in average particle size indicating that H. azteca are able to induce PS-MP fragmentation. This process may facilitate bioaccumulation and trophic particle transfer., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2023 Elsevier B.V. All rights reserved.)

Omics approaches are continuously providing new clues on the mechanisms of action of contaminants in species of environmental relevance, contributing to the emergence of molecular ecotoxicology. Co-expression network approaches represent a suitable methodological framework for studying the rich content of omics datasets. This study aimed to find evidence of key pathways and proteins related to the testicular toxicity in the sentinel crustacean species Gammarus fossarum exposed to endocrine disruptors using a weighted protein co-expression network analysis. From a shotgun proteomics dataset of male gonads of G. fossarum organisms exposed to cadmium (Cd), pyriproxyfen (Pyr) and methoxyfenozide (Met) in laboratory conditions, four distinct modules were identified as significantly correlated to contaminants' exposure. Protein set enrichment analysis identified modules involved in cytoskeleton organization and oxidative stress response associated with the Cd exposure. The module associated with Pyr exposure was associated with endoplasmic reticulum stress (ER) response, and the module correlated with Met exposure was characterized by a significant proportion of amphipod-restricted proteins whose functions are still not characterized. Our results show that co-expression networks are efficient and adapted tools to identify new potential mode of actions from environmental sentinel species, such as G. fossarum, using a proteogenomic approach, even without an annotated genome., (Copyright © 2021. Published by Elsevier B.V.)

Toxicokinetic (TK) models refer to the process of contaminant bioaccumulation as a balance between rate of uptake from different sources (e.g., water or food), and rate of elimination via different processes such as excretion, growth and/or biotransformation. Biotransformation can considerably modify the fate of chemicals in an organism, especially their bioavailability, residence time, and toxicity. Invertebrate models generally neglect this process as they assume a low metabolic activity. However, some species such as Gammarus sp. amphipods are able to metabolize a vast range of organic compounds. Some recent TK models include biotransformation, but they prove limited for estimating related parameters by giving negative values and/or large uncertainties for biotransformation rate(s). Here we propose a generic TK model accounting for biotransformation using a Bayesian framework for simultaneously estimating the parameters. We illustrated the added value of our method by fitting this generic TK model to 22 published datasets of several benthic invertebrate species exposed to different chemicals. All parameters are estimated simultaneously for all datasets and showed narrow estimates. Furthermore, the median model predictions and their 95% credibility intervals showed that the model confidently fitted the data. In most cases the uncertainties around biotransformation rate(s) were reduced in comparison to the original studies. From a methodology standpoint, this paper reflects that Bayesian inference has real added value for simultaneously estimating all TK parameters for parent chemicals and their metabolite(s) based on all available data, while accounting for different types of data and the correlation between parameters. Bayesian inference was able to overcome the limits of previous methods, since no parameters were fixed and no irrelevant negative values were obtained. Moreover, the 95% credibility intervals around model predictions, which are core uncertainties for Environmental Risk Assessment, were easily acquired., (Copyright © 2021 Elsevier Ltd. All rights reserved.)

Antibiotics may constitute a risk for aquatic detritivorous macroinvertebrates (i.e., shredders) via waterborne and dietary antibiotic exposure. In addition, antibiotics can alter the food quality for shredders mediated by shifts in leaf-associated decomposer (i.e., aquatic fungi and bacteria) communities. However, little is known about the relative importance of the waterborne and dietary effect pathway. Therefore, we followed a tiered testing approach aimed at assessing the relative importance of these effect pathways. We employed the antibiotic ciprofloxacin (CIP) and the shredder Gammarus fossarum as model stressor and test species, respectively. In a first step, we assessed the short-term waterborne toxicity of CIP using survival and leaf consumption of G. fossarum as response variables. Alterations in the leaf-associated decomposer community, which may be reflected by their palatability, were assessed using food choice assays. Finally, we conducted a 2 × 2-factorial experiment over 24 days assessing the pathways individually and combined using energy processing (i.e., leaf consumption and feces production), growth and energy storage (i.e., neutral lipid fatty acids) as variables. Short term waterborne exposure indicated low toxicity with LC 50 and EC 50 values of 13.6 and 6.4 mg CIP/L, respectively. At the same time, shredders did not prefer any leaf material during the food choice assay. However, the fungal community was significantly affected in the highest CIP-treatments (0.5 and 2.5 mg/L) suggesting an altered food quality for shredders. This assumption is supported by the results of the long-term assay. At 0.5 mg CIP/L, gammarids' leaf consumption, growth and energy storage were increased when subjected via the dietary pathway, which was linked to changes in the leaf-associated microbial community. Our data highlight the importance of dietary effect pathways for effects on shredders, potentially impacting energy dynamics in detritus-based stream ecosystems., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2020 Elsevier B.V. All rights reserved.)

Aquatic media are ultimate recipients of various contaminants including pesticides pervasively applied in agrosystems. Characterizing the ecotoxicity of pesticides and their mixtures to aquatic wildlife at field-realistic levels is thus crucial for environmental risk assessment. This study aims at assessing the effects of two current-use insecticides, imidacloprid and chlorpyrifos, on Gammarus fossarum using multi-level biomarkers. In microcosms, gammarids were exposed for 72 h to insecticides tested individually or in mixture at 0.01, 0.1 and 1 μg/L of each chemical. Multi-metric responses were assessed at the individual level (behavioural traits: locomotion, respiration and amplexus formation) and the cellular level (enzymes involved in growth, moulting, digestion and cell stress). The results showed insecticide-elicited behavioural and biochemical responses from the lowest concentration of 0.01 μg/L. Overall, single exposures stimulated behavioural traits and inhibited enzymatic activities, highlighting subtle impacts at different organizational levels but these were not dose related. For binary mixtures, antagonistic effects (i.e. less-than-additive) on biomarkers were mainly observed when compared with single exposures. Multi-variable analyses indicated the complementarity of behavioural and biochemical biomarkers in identifying sublethal biological alterations and dose-dependent multiple action sites of insecticides. Besides, the mortality observed only for the mixture at 1 μg/L demonstrated a high lethal potential of insecticides in a simple binary combination. To conclude, this study demonstrates disturbances in individual performances and cellular impairments occurring at environmentally realistic exposure levels in a non-target wild species. Since the sublethal effects, such as those identified with this multi-biomarker approach, could lead to long-term alterations in population dynamics of agricultural areas, they constitute promising early endpoints for risk assessment of insecticides., (Copyright © 2019 Elsevier B.V. All rights reserved.)

High-throughput proteomics can be performed on animal sentinels for discovering key molecular biomarkers signing the physiological response and adaptation of organisms. Ecotoxicoproteomics is today amenable by means of proteogenomics to small arthropods such as Gammarids which are well known sentinels of aquatic environments. Here, we analysed two regional Gammarus pulex populations to characterize the potential proteome divergence induced in one site by natural bioavailable mono-metallic contamination (cadmium) compared to a non-contaminated site. Two RNAseq-derived protein sequence databases were established previously on male and female individuals sampled from the reference site. Here, individual proteomes were acquired on 10 male and 10 female paired organisms sampled from each site. Proteins involved in protein lipidation, carbohydrate metabolism, proteolysis, innate immunity, oxidative stress response and lipid transport were found more abundant in animals exposed to cadmium, while hemocyanins were found in lower abundance. The intrapopulation proteome variability of long-term exposed G. pulex was inflated relatively to the non-contaminated population. These results show that, while remaining a challenge for such organisms with not yet sequenced genomes, taking into account intrapopulation variability is important to better define the molecular players induced by toxic stress in a comparative field proteomics approach., (Copyright © 2019 Elsevier B.V. All rights reserved.)

The amount of pollutants and nutrients entering rivers via point sources is increasing along with human population and activity. Although wastewater treatment plants (WWTPs) greatly reduce pollutant loads into the environment, excess nutrient loading is a problem in many streams. Using a Community and Ecosystem Function (CEF) approach, we quantified the effects of WWTP effluent on the performance of microbes and detritivores associated to organic matter decomposition, a key ecosystem process. We measured organic matter breakdown rates, respiration rates and exo-enzymatic activities of aquatic microbes. We also measured food consumption and growth rates and RNA to body-mass ratios (RNA:BM) of a dominant amphipod Echinogammarus berilloni. We predicted responses to follow a subsidy-stress pattern and differences between treatments to increase over time. To examine temporal effects of effluent, we performed a laboratory microcosm experiment under a range of effluent concentrations (0, 20, 40, 60, 80 and 100%), taking samples over time (days 8, 15 and 30; 4 and 10 replicates to assess microbe and detritivore performance respectively, per treatment and day). This experiment was combined with a field in situ Before-After Control-Impact Paired (BACIP) experiment whereby we added WWTP effluent poured (10 L s -1 during 20-40 min every 2 h) into a stream and collected microbial and detritivore samples at days 8 and 15 (5 and 15 replicates to assess the microbe and detritivore performance respectively, per period, reach and sampling day). Responses were clearer in the laboratory experiment, where the effluent caused a general subsidy response. Field measures did not show any significant response, probably because of the high dilution of the effluent in stream water (average of 1.6%). None of the measured variables in any of the experiments followed the predicted subsidy-stress response. Microbial breakdown, respiration rates, exo-enzymatic activities and invertebrate RNA:BM increased with effluent concentrations. Differences in microbial respiration and exo-enzymatic activities among effluent treatments increased with incubation time, whereas microbial breakdown rates and RNA:BM were consistent over time. At the end of the laboratory experiment, microbial respiration rates increased 156% and RN:BM 115% at 100% effluent concentration. Detritivore consumption and growth rates increased asymptotically, and both responses increased with by incubation time. Our results indicate that WWTP effluent stimulates microbial activities and alters detritivore performance, and stream water dilution may mitigate these effects., (Copyright © 2018 The Authors. Published by Elsevier B.V. All rights reserved.)

Kinetic parameters (uptake from solution and elimination rate constants) of Cu, Ni and Pb bioaccumulation were determined from two Gammarus pulex and three Gammarus fossrum wild populations collected from reference sites throughout France in order to assess the inter-species and the natural inter-population variability of metal bioaccumulation kinetics in that sentinel organism. For that, each population was independently exposed for seven days to either 2.5μgL -1 Cu (39.3nM), 40μgL -1 Ni (681nM) or 10μgL -1 Pb (48.3nM) in laboratory controlled conditions, and then placed in unexposed microcosms for a 7-day depuration period. In the same way, the possible influence of metal exposure history on subsequent metal bioaccumulation kinetics was addressed by collecting wild gammarids from three populations inhabiting stations contaminated either by Cd, Pb or both Pb and Ni (named pre-exposed thereafter). In these pre-exposed organisms, assessment of any changes in metal bioaccumulation kinetics was achieved by comparison with the natural variability of kinetic parameters defined from reference populations. Results showed that in all studied populations (reference and pre-exposed) no significant Cu bioaccumulation was observed at the exposure concentration of 2.5μgL -1 . Concerning the reference populations, no significant differences in Ni and Pb bioaccumulation kinetics between the two species (G. pulex and G. fossarum) was observed allowing us to consider all the five reference populations to determine the inter-population natural variability, which was found to be relatively low (kinetic parameters determined for each population remained within a factor of 2 of the minimum and maximum values). Organisms from the population exhibiting a Pb exposure history presented reduced Ni uptake and elimination rate constants, whereas no influence on Ni kinetic parameters was observed in organisms from the population exhibiting an exposure history to both Ni and Pb. Furthermore Pb bioaccumulation kinetics were unaffected whatever the condition of pre-exposure in natural environment. Finally, these results highlight the complexity of confounding factors, such as metal exposure history, that influence metal bioaccumulation processes and showed that pre-exposure to one metal can cause changes in the bioaccumulation kinetics of other metals. These results also address the question of the underlying mechanisms developed by organisms to cope with metal contamination., (Copyright © 2017 Elsevier B.V. All rights reserved.)

Recently, a protein sequence database was built specifically for the sentinel non-model species Gammarus fossarum using a proteogenomics approach. A quantitative multiplexed targeted proteomics assay (using Selected Reaction Monitoring mass spectrometry) was then developed for a fast and simultaneous quantification of dozens of biomarker peptides specific of this freshwater sentinel crustacean species. In order to assess the relevance of this breakthrough methodology in ecotoxicology, the response patterns of a panel of 26 peptides reporting for 20 proteins from the Gammarus fossarum proteome with putative key functional roles (homeostasis, osmoregulation, nutrition, reproduction, molting,…) were recorded through male and female reproductive cycles and after exposure to environmental concentrations of cadmium and lead in laboratory-controlled conditions. Based on these results, we validated the implication of annotated vtg-like peptides in the oogenesis process, and the implication of Na + /K + ATPase proteins in the molt cycle of organisms. Upon metal (cadmium and lead) contamination, peptides belonging to proteins annotated as involved in antioxidant and detoxification functions, immunity and molting were significantly down-regulated. Overall, this multiplex assay allowed gaining relevant insights upon disruption of different main functions in the sentinel species Gammarus fossarum. This breakthrough methodology in ecotoxicology offers a valid and high throughput alternative to currently used protocols, paving the way for future practical applications of proteogenomics-derived protein biomarkers in chemical risk assessment and environmental monitoring., (Copyright © 2017 Elsevier B.V. All rights reserved.)

For in situ remediation of groundwater contaminated by halogenated hydrocarbons Carbo-Iron ® , a composite of microscale activated carbon and nano Fe 0 , was developed. Against the background of intended release of Carbo-Iron into the environment in concentrations in the g/L-range, potential ecotoxicological consequences were evaluated in the present study. The nano Fei 0 in Carbo-Iron acts as reducing agent and is oxidized in aqueous systems by chlorinated solvents, groundwater constituents (e.g. dissolved oxygen) and anaerobic corrosion. As Carbo-Iron is generally oxidized rapidly after application into the environment, the oxidized state is environmentally most relevant, and Carbo-Iron was used in its oxidized form in the ecotoxicological tests. The amphipod Hyalella azteca was selected as a surrogate test species for functionally important groundwater crustaceans. Effects of Carbo-Iron on H. azteca were determined in a 10-d acute test, a 7-d feeding activity test and a 42-d chronic test. Additionally, a 56-d life cycle test was performed with a modified design to further evaluate effects of Carbo-Iron on adult H. azteca and their offspring. The size of Carbo-Iron particles in stock and test suspensions was determined via dynamic light scattering. Potential uptake of particles into test organisms was investigated using transmission and scanning electron microscopy. At the termination of the feeding and acute toxicity test (i.e. after 7 and 10 d of exposure, respectively), Carbo-Iron had a significant effect on the weight, length and feeding rate of H. azteca at the highest test concentration of 100mg/L. While an uptake of Carbo-Iron into the gut was observed, no passage into the surrounding tissue was detected. In both chronic tests, the number of offspring was the most sensitive endpoint and significant effects were recorded at concentrations ≥50mg/L (42-d experiment) and ≥12.5mg/L (56-d experiment). Parental exposure to oxidized Carbo-Iron significantly exacerbated the acute effects of the nanocomposite on the subsequent generation of H. azteca by a factor >10. The present study indicates risks for groundwater species at concentrations in the mg/L range. Carbo-Iron may exceed these effect concentrations in treated aquifers, but the presence of the pollutant has most likely impaired the quality of this habitat already. The benefit of remediation has to be regarded against the risk of ecological consequences with special consideration of the observed increasing sensitivity of juvenile H. azteca., (Copyright © 2016 Elsevier B.V. All rights reserved.)

Cerium nanoparticles (nCeO2) are widely used in everyday products, as fuel and paint additives. Meanwhile, very few studies on nCeO2 sublethal effects on aquatic organisms are available. We tried to fill this knowledge gap by investigating short-term effects of nCeO2 at environmentally realistic concentrations on two freshwater invertebrates; the amphipod Gammarus roeseli and the bivalve Dreissena polymorpha, using an integrated multibiomarker approach to detect early adverse effects of nCeO2 on organism biology. Differences in the behaviour of the organisms and of nanoparticles in the water column led to differential nCeO2 bioaccumulations, G. roeseli accumulating more cerium than D. polymorpha. Exposure to nCeO2 led to decreases in the size of the lysosomal system, catalase activity and lipoperoxidation in mussel digestive glands that could result from nCeO2 antioxidant properties, but also negatively impacted haemolymph ion concentrations. At the same time, no strong adverse effects of nCeO2 could be observed on G. roeseli. Further experiments will be necessary to confirm the absence of severe nCeO2 adverse effects in long-term environmentally realistic conditions., (Copyright © 2014 Elsevier B.V. All rights reserved.)

Characterizing the exposure routes of an organism and its ability to regulate accumulated contaminants is a crucial step toward developing a biomonitor. To date, very little data are available on the bioaccumulation kinetics of PBDEs in freshwater biota. This study aims at investigating the potential use of a litter-degrader widely distributed in European freshwaters, Gammarus pulex, as an indicator of exposure to PBDEs. In aquatic microcosms, gammarids were exposed to a mixture of brominated congeners (BDE-28, 47, 66, 85, 99, 100, 153, 154 and 183) to assess their ability to bioconcentrate PBDEs. Results show that all tested congeners are highly internalized by G. pulex and uptake rates of PBDEs are closely related to their partition coefficients (Kow). The determination of the elimination rate of BDE-47, the congener most readily accumulated by gammarids, indicated that metabolism and excretion of this congener are low in G. pulex, which argues in favor of its use as a quantitative biomonitor. Finally, bioaccumulation experiments were performed using contaminated leaves to determine the relative importance of dietary uptake in the contamination of gammarids. Even though water is the preeminent exposure route, a significant uptake of BDE-47 through food was observed (27%). We propose a biodynamic model that takes into account both exposure routes to describe BDE-47 bioaccumulation. This study supports the use of this ubiquitous amphipod as an early warning monitor of the bioavailable contamination of freshwaters by PBDEs., (Copyright © 2014 Elsevier B.V. All rights reserved.)

Next generation sequencing using Roche's 454 pyrosequencing platform can be used to generate genomic information for non-model organisms, although there are bioinformatic challenges associated with these studies. These challenges are compounded by a lack of a standardized protocol to either assemble data or to evaluate the quality of a de novo transcriptome. This study presents an assembly of the control and toxicant responsive transcriptome of Melita plumulosa, an Australian amphipod commonly used in ecotoxicological studies. RNA was harvested from control amphipods, juvenile amphipods, and from amphipods exposed to either metal or diesel contaminated sediments. This RNA was used as the basis for a 454 based transcriptome sequencing effort. Sequencing generated 1.3 million reads from control, juvenile, metal-exposed and diesel-exposed amphipods. Different read filtering and assembly protocols were evaluated to generate an assembly that (i) had an optimal number of contigs; (ii) had long contigs; (iii) contained a suitable representation of conserved genes; and (iv) had long ortholog alignment lengths relative to the length of each contig. A final assembly, generated using fixed-length trimming based on the sequence quality scores, followed by assembly using the MIRA algorithm, produced the best results. The 26,625 contigs generated via this approach were annotated using Blast2GO, and the differential expression between treatments and control was determined by mapping with BWA followed by DESeq. Although the mapping generated low coverage, many differentially expressed contigs, including some with known developmental or toxicological function, were identified. This study demonstrated that 454 pyrosequencing is an effective means of generating reference transcriptome information for organisms, such as the amphipod M. plumulosa, that have no genomic information available in databases or in closely related sequenced species. It also demonstrated how optimization of read filtering protocols and assembly approaches changes the utility of results obtained from next generation sequencing studies, and establishes criteria to determine the quality of a de novo assembly in species lacking a reference genome. This new transcriptomic knowledge provides the genomic foundation for the creation of microarray and qPCR assays, serving as a reference transcriptome in future RNAseq studies, and allowing both the biology and ecotoxicology of this organism to be better understood. This approach will allow genomics-based methodology to be applied to a wider range of environmentally relevant species., (Crown Copyright © 2013. Published by Elsevier B.V. All rights reserved.)

The expression of the yolk protein vitellogenin (Vtg) has been used as a biomarker of feminisation in multiple fish species throughout the world. Since the late 1990s, researchers have attempted to develop similar biomarkers to address whether reproductive endocrine disruption also occurs in the males of invertebrate groups such as the Crustacea. To date, the vast majority of studies investigating Vtg induction in male Crustacea have resulted in negative or inconclusive results, leading researchers to question the utility of Vtg expression as a biomarker in this taxon. This study measured the expression of Vtg genes in two intersex phenotypes (termed internal and external) found in the male amphipod, Echinogammarus marinus, and compared them with those of normal males and females. Males presenting the external intersex phenotype are infected with known feminising parasites and display a variety of feminised traits including oviduct structures on their testes and external female brood plates (oostegites). The internal intersex male phenotype, that displays a pronounced oviduct structure on the testes without the external intersex characteristics, is not parasite infected and it is thought to be a result of environmental contamination. Given their morphology, these phenotypes might be considered highly 'feminised' or 'de-masculinised' and can be utilised to test the suitability of feminisation biomarkers. The E. marinus transcriptome was searched for genes resembling Vtg and two sequences were revealed, that we subsequently refer to as Vtg1 and Vtg2. Results from a high-throughput transcriptomic sequencing screen of gonadal cDNA libraries suggested that very low expression (in this manuscript gene transcription is taken to represent gene expression, although it is acknowledged that in addition to transcription, translation, transcript processing, mRNA stability and protein stability can regulate gene expression) of Vtg1 and Vtg2 in normal males (ESTs=1 and 0 for Vtg1 and Vtg2, respectively), internal intersex males (ESTs=0 for both Vtg sequences) and external intersex males (ESTs=5 and 0 for Vtg1 and Vtg2, respectively). In contrast, the sequencing suggested notable levels of expression of both Vtg genes in females (ESTs=1133 and 84 for Vtg1 and Vtg2, respectively). Subsequent qPCR analysis validates these expression levels, with the signal for Vtg1 and Vtg2 transcripts in all male phenotypes being indistinguishable from that caused by contamination of trace levels of genomic DNA or the low-level amplification non-target sequences. These findings suggest that Vtg expression is not notably induced in highly feminised amphipods and is therefore not an appropriate biomarker of feminisation/de-masculination in crustaceans. We discuss our findings in the context of previous attempts to measure Vtg in male crustaceans and suggest a requirement for more appropriate taxon-specific biomarkers to monitor feminisation in these groups., (Copyright © 2013 Elsevier B.V. All rights reserved.)

Estuarine and marine sediments are a probable end point for many engineered nanoparticles (ENPs) due to enhanced aggregation and sedimentation in marine waters, as well as uptake and deposition by suspension-feeding organisms on the seafloor. Benthic infaunal organisms living in sediments encounter relatively high concentrations of pollutants and may also suffer toxic effects of ENPs. We tested whether three heavily used metal oxide ENPs, zinc oxide (ZnO), copper oxide (CuO), and nickel oxide (NiO) were toxic to an estuarine amphipod, Leptocheirus plumulosus. We used results from 10-day laboratory bioassays to estimate potential demographic impacts of ENP exposure. We also evaluated fate and transport pathways of the ENPs in the experiments to elucidate routes of uptake and exposure. Dissolved Zn was found in sediment pore water and overlying water samples at 10 fold the concentrations of Cu or Ni, a pattern indicative of the relatively high dissolution rate of ZnO ENPs compared with CuO and NiO ENPs. Accumulation of metals in amphipod tissues increased with exposure concentrations for all three ENPs, suggesting possible exposure pathways to higher taxa. Amphipods accumulated ≤600 μg Zn and Cu g(-1) and 1000 μg Ni g(-1). Amphipod mortality increased with ZnO and CuO concentrations, but showed no significant increase with NiO to concentrations as high as 2000 μg g(-1). The median lethal concentration in sediment (LC50) of ZnO was 763 μg g(-1) and 868 μg g(-1) for CuO ENPs. Our results indicate that ZnO and CuO ENPs, but not NiO ENPs, are toxic to L. plumulosus and that ZnO toxicity primarily results from Zn ion exposure while CuO toxicity is due to nanoparticle exposure., (Copyright © 2013 Elsevier B.V. All rights reserved.)

Differences between the bioavailability of cadmium in a periphyton diet and an artificial laboratory diet (TetraMin(®)) have important consequences for predicting bioaccumulation and toxicity in the freshwater amphipod Hyalella azteca. The assimilation efficiency (AE) of Cd was compared between periphyton and TetraMin(®) at low (1510 and 358 nmol/g ash-free dry mass respectively) and chronically lethal (31,200 and 2890 nmol/g ash-free dry mass respectively) Cd concentrations and in fresh and dry forms using a (109)Cd radiotracer pulse-chase feeding technique. Assimilation efficiency of Cd from periphyton (AE=3-14%) was lower than that for TetraMin(®) (AE=44-86%) regardless of Cd concentration or food form. Ingestion rate (IR) was lower for dry than fresh forms of periphyton (0.042 and 0.16 g AFDM/g H. azteca/day respectively) and TetraMin(®) (0.19 and 0.87 AFDM/g H. azteca/day respectively) and depuration rate (k(e)) did not differ statistically with food type, form or Cd concentration (0.032-0.094 d(-1)). Biokinetic models with parameters of AE, IR and k(e) were used to estimate bioaccumulation from the separate food types. These estimates were compared to those from an independent chronic Cd saturation bioaccumulation model. While the model estimates did not concur, a sensitivity analysis indicated that AE and IR were the most influential biokinetic model parameters for Cd in periphyton and TetraMin(®) respectively. It was hypothesized that AE was underestimated for Cd in periphyton due to a non-adapted gut enzyme system and IR was overestimated for Cd in TetraMin(®) due to an initial rapid ingestion phase in H. azteca's feeding habits. This research demonstrated the importance of using ecologically relevant food types in laboratory experiments and verifying acute biokinetic model predictions of dietary metal contribution with those derived from a chronic exposure which is more representative of a field exposure scenario., (Crown Copyright © 2012. Published by Elsevier B.V. All rights reserved.)

A vitellogenin (Vg) mass spectrometry-based assay was recently developed to actively biomonitor and assess the exposure of the amphipod Gammarus fossarum to endocrine-disrupting chemicals in freshwater hydrosystems. This paper focuses on the appropriate use of this biomarker, which requires good knowledge of its basal level in males and its natural variability related to intrinsic biotic and environmental abiotic factors. To obtain the lowest biomarker variability, we first studied some of these confounding factors. We observed that the spermatogenesis stage did not have an impact on the Vg level, allowing flexibility in the choice of transplanted gammarids. In the second part of the study, males were transplanted in two clean stations for 21 days, with results indicating a spatial and temporal variability of Vg levels. These Vg changes could not be correlated to environmental factors (e.g., temperature, pH and hardness of waters). Vg induction was then assessed in 21 stations having various levels of contamination. Inductions were observed for only two of the impacted stations studied. Under reference and contaminated conditions, a high interindividual variability of Vg levels was observed in caged organisms, severely limiting the sensitivity of the biomarker and its ability to detect a significant endocrine-disruptor effect. This may be explained by unidentified environmental factors that should later be determined to improved the use of Vg as a biomarker in male G. fossarum. Moreover, as discussed in this paper, recent advancements regarding the pleiotropic functions of the Vg gene in some species may complicate the application of this biomarker in males of invertebrate species., (Copyright © 2012 Elsevier B.V. All rights reserved.)

This work focused on the validation of biological specificity of the quantitative LC-MS/MS assay by checking the natural variability of Vg levels during the reproductive cycle in Gammarus fossarum (i.e., including oogenesis and embryogenesis). Laboratory tests were performed for 21 days under controlled conditions to assess Vg changes in male and female gammarids after exposure to chemical stress. Females were exposed to two crustacean hormones, 20-hydroxyecdysone (0.01, 1 and 100 μg L⁻¹) and methyl-farnesoate (0.01, 1 and 100 μg L⁻¹). No effect was recorded for 20-hydroxyecdysone, whereas in females exposed to methyl-farnesoate a deleterious impact on Vg production was observed. Males were exposed to crustacean hormones 20-hydroxyecdysone (0.01, 1 and 100 μg L⁻¹) and methyl-farnesoate (0.01, 1 and 100 μg L⁻¹), the insecticide methoxyfenozide (0.001, 0.1 and 10 μg L⁻¹), the fungicide propiconazole (0.001, 0.1, 10 and 1000 μg L⁻¹), and the pharmaceutical products benzophenone, carbamazepine, cyproterone, and R-propranolol (0.001, 0.1, 10 and 1000 μg L⁻¹). Induction of Vg synthesis was recorded in males exposed to cyproterone, methoxyfenozide, methyl-farnesoate, and propiconazole. Finally, we validated the function of the ILIPGVGK peptide used to track vitellogenin in G. fossarum across reproductive processes (vitellogenesis and embryogenesis), and results confirmed the energy reserve role of Vg during embryo development. We show that oocyte surface measurement is directly related to Vg levels in the oocyte, constituting a reliable indicator of egg quality in G. fossarum. Consequently, it could be used as a reliable tool for biomonitoring programs. We recorded induction of Vg in male G. fossarum; however, the possible use of this tool as a specific biomarker of exposure to endocrine disruption should be confirmed in further studies., (Copyright © 2012 Elsevier B.V. All rights reserved.)

We use published data on the different patterns of the bioaccumulation of zinc by three crustaceans, the caridean decapod Palaemon elegans, the amphipod Orchestia gammarellus and the barnacle Amphibalanus amphitrite, to construct comparative biodynamic models of the bioaccumulation of zinc into metabolically available and detoxified components of accumulated zinc in each crustacean under both field and laboratory toxicity test conditions. We then link these bioaccumulation models to the onset of toxic effects on exposure of the crustaceans to high dissolved zinc bioavailabilities, using the tenets that toxicity effects are related to the total uptake rate of the toxic metal, and that toxicity is not usually dependent on the total accumulated metal concentration but always on the concentration of accumulated metal that is metabolically available. We dismiss the general concept that there is a critical accumulated body concentration of a metal in an invertebrate at which toxicity ensues, except under specific circumstances involving a rare lack of storage detoxification of accumulated metal. We thus propose a theoretical framework that can be extended to other metals and other aquatic invertebrates (indeed other animals) to explain the variation in the relationship between bioaccumulated body concentrations and toxicity, and subsequently to predict this relationship in many other species for which we have bioaccumulation modelling data., (Copyright © 2011 Elsevier B.V. All rights reserved.)

We investigated the sensitivity of the freshwater crustacean amphipod Gammarus pulex towards organic xenobiotic compounds in comparison to the sensitivity of the crustacean cladoceran Daphnia magna. In addition we studied the influence of the chemical's mode of action on the relationship between the sensitivity of G. pulex and that of D. magna. We tested the acute toxicity of twelve compounds (Malathion, Aldicarb, Carbofuran, 2,4-dichloroaniline, 2,4-dichlorophenol, 1,2,3-trichlorobenzene, 4,6-dinitro-o-cresol, 2,4,5-trichlorophenol, Ethylacrylate, 4-nitrobenzyl-chloride, Sea-nine, Imidacloprid) with different modes of action and physicochemical properties towards the freshwater amphipod G. pulex in laboratory experiments. Additional toxicity data was collected from the peer-reviewed literature and databases (data pairs for 44 chemicals in total). The chemicals were assigned to seven mode of action groups. The relationship between the sensitivity of G. pulex (48h-LC50s and 96h-LC50s) and that of D. magna (48h-EC50s) was investigated using regression analysis and correlation plots. G. pulex is two to three orders of magnitude more sensitive towards neonicotinoids than D. magna (P=0.0046, n=3). For organophosphates we found that D. magna is more sensitive than G. pulex by approximately a factor of six (P=0.0256, n=6). There was no significant difference between the sensitivity of D. magna and that of G. pulex in any of the other mode of action groups; however chemicals with the same mode of action grouped together in the same area of the correlation plot. Without the neonicotinoids 75% of all G. pulex toxicity data were within one order of magnitude of the D. magna data and 100% within two orders of magnitude. The regressions with all data and with all data minus neonicotinoids were both significant linear relationships with slopes around one and intercept around zero. Thus, G. pulex is generally equally sensitive towards organic xenobiotics as D. magna., (Copyright © 2011 Elsevier B.V. All rights reserved.)

Organisms in marine sediments are usually exposed to mixtures of polycyclic aromatic hydrocarbons (PAHs), whereas risk assessment and management typically focus on the effects of single PAHs. This can lead to an underestimation of risk if the effects of single compounds are additive or synergistic. Because of the virtually infinite number of mixture-combinations, and the many different targeted organisms, it would be advantageous to have a model for the assessment of mixture effects. In this study we tested whether chemical activity, which drives the partitioning of PAHs into organisms, can be used to model the baseline toxicity of mixtures. Experiments were performed with two benthic amphipod species (Orchomonella pinguis and Corophium volutator), using passive dosing to control the external exposure of single PAHs and mixtures of three and four PAHs. The baseline toxicity of individual PAHs at water saturation generally increased with increasing chemical activity of the PAHs. For O. pinguis, the baseline toxicity of PAH mixtures was successfully described by the sum of chemical activities. Some compounds and mixtures showed a delayed expression of toxicity, highlighting the need to adjust the length of the experiment depending on the organism. On the other hand, some of the single compounds had a higher toxicity than expected, possibly due to the toxicity of PAH metabolites. We suggest that chemical activity of mixtures can, and should, be used in addition to toxicity data for single compounds in environmental risk assessment., (Copyright © 2011 Elsevier B.V. All rights reserved.)

The River Hayle, Cornwall, UK exhibits pronounced Cu and Zn concentration gradients which were used to compare the metal handling abilities of two populations of Gammarus pulex (Crustacea: Amphipoda). One population was native to the Hayle region (Drym) and presumably has been historically impacted by elevated Cu and Zn levels, whilst naïve gammarids were collected from the River Cray, Kent, UK. Both populations were subject to a 32 day in situ exposure at four R. Hayle sites (Drym, Godolphin, Relubbus and St. Erth). Mortality (LT50), Cu and Zn accumulation and sub-cellular distribution, and oxidative stress (malondialdehyde production) increased with the expected Cu and Zn bioavailabilities at the four sites (i.e. Godolphin>Relubbus>St. Erth>Drym). The naïve population experienced greater metal induced effects in terms of Cu and Zn accumulation, oxidative stress responses and lower LT50s. Analysis of Cu and Zn sub-cellular distribution, however, revealed no significant differences in metal handling. In both populations each metal was localised predominantly to the sub-cellular fraction containing metal bound to metallothionein-like proteins (MTLP) or that holding both metal-rich granules (MRG) and exoskeleton, MTLP and MRG binding being indicative of metal detoxification. However, a greater capacity for detoxified metal storage is not a mechanism implicated in the perceived tolerance of the historically impacted gammarids. Instead our results suggest that the historically impacted population was adapted for lower uptake of Cu and Zn leading to lower bioaccumulation, stress response and ultimately mortality. These results demonstrate not only the usefulness of the in situ methodology, but also that differences in population exposure history can cause significant differences in metal responses during exposure at higher concentrations., (Copyright © 2011 Elsevier B.V. All rights reserved.)

The effects of serotonin altering parasites, serotonin, the anti-depressant fluoxetine, plus two other highly prescribed pharmaceuticals (carbamazepine and diclofenac) on the behaviour of the marine amphipod, Echinogammarus marinus were investigated. Acanthocephalan parasites are known to alter the swimming behaviour in their amphipod hosts through changes in serotonergic activity resulting in increased predation. Behavioural assays were adapted to record changes in phototaxis and geotaxis behaviour in male E. marinus following 7, 14 and 21 days exposure to serotonin and each pharmaceutical compound at 4 concentrations compared to a control (between 10 ng/L and 10 microg/L). E. marinus infected with acanthocephalans parasites had both significantly higher phototaxis and geotaxis scores than those of uninfected specimens. Phototaxis and geotaxis behaviour increased significantly in a concentration-dependent manner with exposure to serotonin. Fluoxetine significantly altered phototaxis and geotaxis activity in what appeared to be a non-monotonic concentration response curve with the greatest behavioural changes observed at 100 ng/L. The main patterns of these behavioural responses were consistent between two trials and the 3 weeks exposure with specimens spending more time within the light and occurring higher in the water column. No obvious trends could be concluded in the phototaxis and geotaxis scores from individuals exposed to carbamazepine or diclofenac as might be expected from their known mode of action. From this study phototaxis and geotaxis behaviour have been observed to be affected by exposure to serotonin modulators. Parasite studies have shown strong links between changes in behaviour and increased predation risk correlating with changes in serotonergic activity. This study has highlighted the potential for highly prescribed anti-depressant drugs to change the behaviour of an ecologically relevant marine species in ways which could conceivably lead to population level effects., (Copyright 2010 Elsevier B.V. All rights reserved.)

Perfluorooctane sulfonate (PFOS) is the focus of intense toxicity research due to its persistence and widespread occurrence in biota. Studies on benthic invertebrates have shown them to be subjects of high PFOS exposure. However, effects on benthic invertebrates exposed to PFOS in the field are still far from elucidated. To fill a knowledge gap on concentrations and effects in benthic invertebrates, a microcosm study on the benthic amphipod Monoporeia affinis was performed. Field collected M. affinis were analysed for PFOS and showed average background concentrations 39 and 58 ng/g (wet weight) in two different samplings. The field collected animals were exposed to three concentrations of PFOS (50, 200 and 5000 microg PFOS/L water) for 3 weeks during gonad development. Body burdens of PFOS were determined after experiment termination. Results showed negative effects on survival and reproduction effects such as decreased sexual maturation and decreased oocyte viability caused by PFOS exposure. Additionally, a follow-up experiment demonstrated a significant increase in the infection incidence by a microsporidian muscle parasite in animals exposed to PFOS at tissue concentrations in the range of concentrations found in field collected benthic amphipods. This is the first study to demonstrate increased microsporidian infection with pollutant exposure and it suggests that ecologically relevant PFOS concentrations could be sufficient to elicit these effects., (Copyright (c) 2010 Elsevier B.V. All rights reserved.)

There has been a growing interest in establishing how the sub-cellular distribution of metals in macro-invertebrate prey affects metal trophic bioavailability and toxicity. In this study, the crustacean Gammarus pulex was exposed to 300mugCdl(-1) spiked with (109)Cd for 13 days, from which the two principal metal containing sub-cellular fractions, the metallothionein-like protein (MTLP) and the metal rich granule and exoskeleton (MRG+exo) were isolated. These fractions were produced at equal metal content, incorporated into gelatin and fed to zebrafish as a single meal; assimilation efficiency (AE), carcass and gut tissue metal concentrations and gut lipid peroxidative damage measured as malondialdehyde (MDA) were assessed. The AE of cadmium bound to the MTLP fraction was 32.1+/-5.6% which was significantly greater than the AE of MRG+exo bound Cd, 13.0+/-2.1% (p<0.05). Of the metal retained by the fish at 72h post-feeding, 94% of MTLP-Cd had been incorporated into the carcass, whereas a significant proportion (46%) of MRG+exo-Cd, although assimilated, appeared to remain associated with intestinal tissue. However, this did not translate into a gut tissue concentration difference with 6.8+/-1.2ngCdg(-1) in fish fed MTLP-Cd compared to 9.5+/-1.4ngCdg(-1) in fish fed MRG+exo fraction. Both feeds led to significantly increased MDA levels compared to the control group (gelatin only feed), but MRG+exo feed caused significantly more oxidative damage than the MTLP feed (p<0.01). Thus, MTLP-Cd is more bioavailable than the cadmium bound to granules and exoskeleton, but it was the latter fraction, largely considered as having limited bioavailability, that appeared to exert a greater localised oxidative injury to the digestive tract of zebrafish., (Copyright (c) 2009 Elsevier B.V. All rights reserved.)

Polar marine surface waters are characterized by high levels of dissolved oxygen, seasonally intense UV irradiance and high levels of dissolved organic carbon. Therefore, the Arctic sea-ice habitat is regarded as a strongly pro-oxidant environment, even though its significant ice cover protects the ice-associated (=sympagic) fauna from direct irradiation to a large extent. In order to investigate the level of resistance to oxyradical stress, we sampled the sympagic amphipod species Gammarus wilkitzkii during both winter and summer conditions, as well as exposed specimens to simulated levels of near-natural and elevated levels of UV irradiation. Results showed that this amphipod species possessed a much stronger antioxidant capacity during summer than during winter. Also, the experimental UV exposure showed a depletion in antioxidant defences, indicating a negative effect of UV exposure on the total oxyradical scavenging capacity. Another sympagic organism, Onisimus nanseni, was sampled during summer conditions. When compared to G. wilkitzkii, the species showed even higher antioxidant scavenging capacity.

Chronic exposure to polluted field conditions can impact metal bioavailability in prey and may influence metal transfer to predators. The present study investigated the assimilation of Cd, Hg and organic carbon by grass shrimp Palaemonetes pugio, collected along an impact gradient within the New York/New Jersey Harbor Estuary. Adult shrimp were collected from five Staten Island, New York study sites, fed (109)Cd- or (203)Hg-labeled amphipods or (14)C-labeled meals and analyzed for assimilation efficiencies (AE). Subsamples of amphipods and shrimp were subjected to subcellular fractionation to isolate metal associated with a compartment presumed to contain trophically available metal (TAM) (metal associated with heat-stable proteins [HSP - e.g., metallothionein-like proteins], heat-denatured proteins [HDP - e.g., enzymes] and organelles [ORG]). TAM-(109)Cd% and TAM-(203)Hg% in radiolabeled amphipods were approximately 64% and approximately 73%, respectively. Gradients in AE-(109)Cd% ( approximately 54% to approximately 75%) and AE-(203)Hg% ( approximately 61% to approximately 78%) were observed for grass shrimp, with the highest values exhibited by shrimp collected from sites within the heavily polluted Arthur Kill complex. Population differences in AE-(14)C% were not observed. Assimilated (109)Cd% partitioned to the TAM compartment in grass shrimp varied between approximately 67% and approximately 75%. (109)Cd bound to HSP in shrimp varied between approximately 15% and approximately 47%, while (109)Cd associated with metal-sensitive HDP was approximately 17% to approximately 44%. Percentages of assimilated (109)Cd bound to ORG were constant at approximately 10%. Assimilated (203)Hg% associated with TAM in grass shrimp did not exhibit significant variation. Percentages of assimilated (203)Hg bound to HDP ( approximately 47%) and ORG ( approximately 11%) did not vary among populations and partitioning of (203)Hg to HSP was not observed. Using a simplified biokinetic model of metal accumulation from the diet, it is estimated that site-specific variability in Cd AE by shrimp and tissue Cd burdens in field-collected prey (polychaetes Nereis spp.) could potentially result in up to approximately 3.2-fold differences in the dose of Cd assimilated by shrimp from a meal in the field. The results of this study also suggest that chronic field exposure can impact mechanisms of metal transport across the gut epithelium that do not influence carbon assimilation. Differences in the assimilation and subcellular partitioning of metal may have important implications for metal toxicity in impacted shrimp populations.

The majority of metabolomic studies used in ecotoxicology have implemented (1)H NMR analysis. Despite constant improvement, major limitations of NMR-based techniques include relatively low sensitivity that results in an examination of a limited number of metabolites. An alternative approach is the use of liquid or gas chromatography (GC) for separation of metabolites and mass spectrometry (MS) for their quantification and identification. The objective of our study was to develop a two dimensional GC coupled with time of flight MS (GCxGC/TOF-MS) coupled with multivariate analysis to compare metabolite profiles of Diporeia under different environmental conditions. We compared metabolite profiles between Diporeia collected from Lake Michigan (declining populations) to those residing in Lake Superior (stable populations), and also between Diporeia exposed to a chemical stressor (atrazine) and controls. Overall, 76 and 302 total metabolites were detected from the lake comparison and atrazine studies, respectively. Many of the identified metabolites included fatty acids, amino acids, and hydrocarbons. Furthermore, we observed unique and almost non-overlapping metabolite profiles in both studies. In conclusion, we established the feasibility of using GCxGC/TOF-MS for detecting metabolites as well as developed software to align and merge chromatographic peaks to compare metabolite differences between invertebrate groups sampled under different environmental conditions. This ability to detect unique metabolite profiles under different environmental conditions will increase our undertsanding on the physiological processes and whole-organism reponses occuring as a result of exposure to different environmental stressors.

Data on the toxicity of uranium in sediments to Hyalella azteca and the effect of overlying water chemistry are limited. This study exposed H. azteca to sediments spiked with U (0-10,000 microg U/g dry weight) and five different overlying waters, which varied independently in hardness and alkalinity. Water pH had a major effect on U bioavailability and uptake by H. azteca. Uranium toxicity was higher when overlying water pH was low, while desorption of U into the overlying water increased with increasing pH. There appears to be little effect of Ca on U uptake, other than its influence on U speciation. Experiments with caged animals indicate that U accumulation and toxicity occur mainly through the dissolved phase rather than the solid phase. Uranium bioaccumulation is a more reliable indicator of U toxicity than U concentration in water or sediment. Uranium bioaccumulation in the H. azteca and U adsorption to sediment can be satisfactorily explained using saturation models.

Mixtures were produced of "equi-toxic" concentrations of 10 elements at the 4-week LC25 for Hyalella azteca. Bioaccumulation was determined in 1-week exposures. The first mixtures tested included seven elements; As, Cd, Co, Cr, Ni, Pb and Tl. Copper, Mn and Zn were not included in the initial tests due to potential confounding effects, such as regulation of Cu and Zn by H. azteca and the high concentrations of Mn required to be "equi-toxic", which might cause adsorption of metals to Mn hydroxides if these were formed. The second set of tests included the seven element mixture in combination with, Cu, Mn and Zn individually, the binary pairs, Cu-Mn, Cu-Zn and Mn-Zn; and the tertiary group Cu, Mn and Zn. Interaction factors (IF) were computed which quantified each element's impact on the bioaccumulation of the other nine. Cobalt, Cd and Ni bioaccumulation was significantly inhibited with increasing number of metals in the mixture. Arsenic bioaccumulation was enhanced with increasing number of metals in the mixture exposure. Lead bioaccumulation was enhanced by some mixture combinations. Bioaccumulation of Cr, Cu, Mn, Tl and Zn were not significantly affected by exposure to other metals.

We studied cellular energy allocation (CEA) in three Arctic benthic species (Gammarus setosus (Amphipoda), Onisimus litoralis (Amphipoda), and Liocyma fluctuosa (Bivalvia)) exposed to oil-related compounds. The CEA biomarker measures the energy budget of organisms by biochemically assessing changes in energy available (carbohydrates, protein and lipid content) and the integrated energy consumption (electron transport system activity (ETS) as the cellular aspect of respiration). Energy budget was measured in organisms subjected to water-accommodated fraction (WAF) of crude oil and drill cuttings (DC) to evaluate whether these compounds affect the energy metabolism of the test species. We observed significantly lower CEA values and higher ETS activity in G. setosus subjected to WAF treatment compared to controls (por=0.19). Different responses to oil-related compounds between the three test species are likely the result of differences in feeding and burrowing behavior and species-specific sensitivity to petroleum-related compounds.

Dissolved organic matter (DOM) is known to reduce the bioavailability of metals in aquatic systems. This study evaluated the effects of DOM from various sources (e.g., Little Bear Lake sediment, Suwannee River, peat moss) and various DOM fractions (humic acids, HA; fulvic acids, FA) on the bioavailability of nickel (Ni) to Hyalella azteca, a common freshwater benthic invertebrate. In particular, this study was conducted to evaluate the effect of surficial sediment DOM on Ni bioavailability. Short-term (48 h) acute toxicity tests with H. azteca conducted in synthetic water demonstrated that the aqueous Ni concentrations required for lethality were greater than what could be significantly complexed by environmentally relevant concentrations of dissolved organic carbon (DOC: 0.6-30.4 mg/L). At Ni concentrations sublethal to H. azteca (500 microg/L), the bioavailability of Ni was significantly reduced in the presence of representative surface water DOC concentrations regardless of DOC source or fraction. DOC fraction (i.e., FA and HA) differentially affected Ni speciation, but had little or no effect on Ni accumulation by H. azteca. Tissue Ni was found to be strongly dependent upon the Ni(2+) concentration in the exposure solutions and the Ni:DOC ratio. Overall, the concentration of DOC played a greater role than either DOC source or fraction in determining Ni speciation and hence bioavailability and toxicity to H. azteca.

Pyrethroid insecticides are known for their potential toxicity to aquatic invertebrates and many fish species. A significant problem in the study of pyrethroid toxicity is their extreme hydrophobicity. They can adsorb to test container surfaces and many studies, therefore, report pyrethroid levels as nominal water concentrations. In this study, pyrethroid adsorption to sampling and test containers was measured and several container treatments were examined for their ability to decrease pyrethroid adsorption. None of the chemical treatments were successful at preventing pyrethroid loss from aqueous samples, but vortexing of containers served to resuspend pyrethroids. The effects of the observed adsorption on Ceriodaphnia dubia and Hyalella azteca permethrin toxicity were examined. Species-specific results showed a time-dependent decrease in toxicity following pyrethroid adsorption to test containers for C. dubia, but not for H. azteca. These results demonstrate that pyrethroid adsorption to containers can significantly affect the observed outcome in toxicity-testing and serves as a caution for researchers and testing laboratories.

The effect of major ions (Ca, Mg, Na, and K) and pH on Cu toxicity (LC50) to Hyalella azteca was determined in 1 week exposures. The simplest equation for describing Cu toxicity is a linear relationship between the total dissolved Cu LC50 and Ca and Na in water, ignoring pH. This equation would be useful in tier one of a two-tiered approach; if the measured dissolved Cu exceeds the value predicted from the equation, the sample should either be tested for toxicity, or a more detailed chemical speciation analysis can be conducted. The data were not consistent with a single-binding-site biotic ligand model, assuming that toxicity was due to the free Cu ion alone. However, toxicity could be predicted using a two-binding-site model. This requires separate coefficients to account for the effects of Ca and Na at low and high pH values (6.5-8.4), corresponding to the different binding sites (Mg and K did not affect toxicity). The single-binding-site BLM does not allow for this. Toxicity of Cu hydroxide or carbonate complexes does not need to be invoked, but cannot be excluded, and several models invoking the toxicity of these complexes can also explain the data. The free ion LC50 is strongly dependent on pH, but the LC50 for total dissolved Cu is almost pH independent. The effects of Ca and Na on the free ion LC50 are very different at high and low pH (contrary to single-site biotic ligand model predictions), but similar for total dissolved Cu. Published data suggest that the same model, with different coefficients, can also be applied to Daphnia and fish. A more critical evaluation of the effects of cations at both low and high pH for organisms other than Hyalella is needed to determine if the BLM needs to be adjusted to incorporate more than one binding site for other species as well. Hydrogen ions reduce the toxicity of free Cu ions to Hyalella, but Cu also reduces the toxicity of hydrogen ions. A mixture model accounting for the joint toxicity of Cu and pH, as well as their mutual antagonistic effects, is presented.

Bioaccumulation of Cd, Pb, Cu and Zn in the Antarctic gammaridean amphipod Paramoera walkeri (Stebbing, 1906) was investigated at Casey station (Australian Antarctic Territory). The main goals were to provide information on accumulation strategies of the organisms tested and to verify toxicokinetic models as a predictive tool. The organisms accumulated metals upon exposure and it was possible to estimate significant model parameters of two-compartment and hyperbolic models. These models were successfully verified in a second toxicokinetic study. However, the application of hyperbolic models appears to be more promising as a predictive tool for metals in amphipods compared to compartment models, which have failed to adequately predict metal accumulation in experiments with increasing external exposures in previous studies. The following kinetic bioconcentration factors (BCFs) for the theoretical equilibrium were determined: 150-630 (Cd), 1600-7000 (Pb), 1700-3800 (Cu) and 670-2400 (Zn). We find decreasing BCFs with increasing external metal dosing but similar results for treatments with and without natural UV radiation and for the combined effect of different exposure regimes (single versus multiple metal exposure) and/or the amphipod collective involved (Beall versus Denison Island). A tentative estimation showed the following sequence of sensitivity of P. walkeri to an increase of soluble metal exposure: 0.2-3.0 microg Cd l(-1), 0.12-0.25 microg Pb l(-1), 0.9-3.0 microg Cu l(-1) and 9-26 microg Zn l(-1). Thus, the amphipod investigated proved to be more sensitive as biomonitor compared to gammarids from German coastal waters (with the exception of Cd) and to copepods from the Weddell Sea inferred from literature data.