Reduced bioavailability of Au and isotopically enriched 109Ag nanoparticles transformed through a pilot wastewater treatment plant in Hyalella azteca under environmentally relevant exposure scenarios.

Wastewater Treatment Plants (WWTP) are a major repository and entrance path of nanoparticles (NP) in the environment and hence play a major role in the final NP fate and toxicity. Studies on silver nanoparticles (AgNP) transport via the WWTP system and uptake by aquatic organisms have so far been carried out using unrealistically high AgNP concentrations, unlikely to be encountered in the aquatic environment. The use of high AgNP concentrations is necessitated by both the low sensitivity of the detection methods used and the need to distinguish background Ag from spiked AgNP. In this study, isotopically enriched 109AgNP were synthesized to overcome these shortcomings and characterized by a broad range of methods including transmission electron microscopy, dynamic and electrophoretic light scattering. 109AgNP and gold NP (AuNP) were spiked to a pilot wastewater treatment plant fed with municipal wastewater for up to 21 days. AuNP were used as chemically less reactive tracer. The uptake of the pristine and transformed NP present in the effluent was assessed using the benthic amphipod Hyalella azteca in fresh- and brackish water exposures at environmentally relevant concentrations of 30 to 500 ng Au/L and 39 to 260 ng Ag/L. The unique isotopic signature of the 109AgNP allowed to detect the material at environmentally relevant concentrations in the presence of a much higher natural Ag background. The results show that the transformations reduce the NP uptake at environmentally relevant exposure concentrations. For 109Ag, lower accumulation factors (AF) were obtained after exposure to transformed NP (250-350) compared to the AF values obtained for pristine 109AgNP (750-840). The reduced AF values observed for H. azteca exposed to effluent from the AuNP-spiked WWTP indicate that biological transformation processes (e.g. eco-corona formation) seem to be involved in addition to chemical transformation.

Improving Sediment Toxicity Testing for Very Hydrophobic Chemicals: Part 2-Exposure Duration, Upper Limit Test Concentrations, and Distinguishing Actual Toxicity from Physical Effects.

Sediment toxicity testing with very hydrophobic organic chemicals (VHOCs) is challenging because of the chemicals' low aqueous solubilities and slow kinetics. The present study presents the results of experiments investigating whether the standard exposure duration of 28 days with benthic invertebrates is sufficient for VHOCs; above which concentrations in sediment VHOCs are present as "free phase," that is, crystals or non-aqueous-phase liquids (NAPLs); and whether it is possible to discriminate between actual VHOC toxicity and physical effects caused by NAPLs through fouling of the test organisms. The results suggest that the standard sediment toxicity test duration is sufficient for obtaining steady-state VHOC concentrations in Hyalella azteca and Lumbriculus variegatus, provided that spiking and equilibration are performed properly (i.e., no free phase present). Under these conditions, transient (days 3-20) peak-shaped toxicokinetics were observed, with steady-state concentrations reached at approximately 28 days. The concentration above which NAPLs are present, the so-called critical separate phase concentration (CSPC), was determined for several VHOCs by modeling and two experimental methods. Modeling resulted in unrealistic and variable data and therefore should be applied with caution. Experimentally determining CSPCs was successful and yielded values of approximately 1000 (400-2000) mg/kg dry weight, depending on the chemical. Finally, it was demonstrated that distinguishing actual toxicity from physical effects is possible by applying a well-considered test setup, combining toxicity tests with multiple invertebrates (including Lumbriculus, which serves as a negative control for fouling); a broad test concentration range, preferably up to at least 30 000 mg/kg; and passive sampling to localize the CSPC. Applying this setup, false-positive effects due to fouling, as well as false-negative results due to testing at too low concentrations (trying to stay below the CSPC), can be avoided. Environ Toxicol Chem 2024;43:1728-1739. © 2023 The Authors. Environmental Toxicology and Chemistry published by Wiley Periodicals LLC on behalf of SETAC.

Two new species of Hyalella (Amphipoda, Dogielinotidae) from the Humid Chaco ecoregion of Paraguay.

The freshwater amphipod genus Hyalella Smith, 1874 is widely distributed in the Neotropics, with several biogeographically restricted species and a high cryptic diversity throughout South America. Tens of species of Hyalella have been documented from nearby Brazil and Argentina, but no systematic record of the genus exists for Paraguay. Here we describe two new species of Hyalella: H.mboituisp. nov. and H.juliasp. nov. from the Ñeembucú wetlands of southwestern Paraguay. Hyalellamboituisp. nov. and H.juliasp. nov. are characterised by a dorsally smooth body, pigmented eyes, uropod 1 endopod with a curved seta, the dorsal margin of uropod 3 ramus without setae, and uropod 3 peduncle longer than wide and with six setae apically. The two species are distinguished by their diagnostic mouthparts, with a median serrated edge on the lacinia mobilis in H.mboituisp. nov. and two elongated lateral denticles with a serrated edge in H.juliasp. nov., and by the presence of a pronounced cup for the dactylus on gnathopod 2 in H.mboituisp. nov. In addition, they show differences in the number of articles on antennae 1 and 2, in the relative length of the pereiopods, and in the numbers and types of setae on their gnathopods and uropods 1-3. Hyalellamboituisp. nov. and H.juliasp. nov. represent the first taxonomically documented occurrence of Paraguayan freshwater amphipods. These new taxa attest to the largely unmapped species richness of freshwater invertebrates in the Humid Chaco of Paraguay. This potential biodiversity hotspot is currently under threat from land conversion, highlighting the need for more systematic studies and effective conservation of the local invertebrate biodiversity.

Avoidance behavior of Hyalella azteca in response to three common-use insecticides.

Non-target organisms in aquatic environments may experience lethal or sublethal effects following exposure to contaminants. Most protocols and regulations, however, are designed to provide protection from lethal effects and are thus based on conventional estimates of population lethality. The relative lack of reliable behavioral endpoints makes it challenging to implement regulations that are similarly protective against sublethal toxicity. The objective of this study was to quantify the avoidance behavior of Hyalella azteca when exposed to three insecticides-bifenthrin (B), chlorpyrifos (C), and permethrin (P)-at a range of estimated lethal concentrations. A two-choice behavioral arena was used for each chemical to quantify H. azteca activity and time spent in either uncontaminated sediment or sediment spiked at concentrations reflecting estimated 48-h lethal concentrations (LC50, LC25, and LC10). For all three insecticides, naïve H. azteca demonstrated a preference for the uncontaminated sediment over the contaminated sediment at the LC50 (B: 312 ng/gOC; C: 1265 ng/gOC; P: 5042 ng/gOC) and LC25 (B: 230 ng/gOC; C: 859 ng/gOC; P: 3817 ng/gOC), spending significantly more time in the uncontaminated side of the arena. H. azteca did not avoid sediment at LC10 (B: 204 ng/gOC; C: 609 ng/gOC; P: 1515 ng/gOC) levels, indicating the existence of a potential threshold of detection. Despite the lack of substrate preference at this exposure level, H. azteca were nevertheless more active (i.e., increased zone-switching) when exposed to bifenthrin at the LC10, suggesting a possible irritation response (e.g., movement after exposure) to this chemical. Our results provide evidence that H. azteca exhibit innate avoidance responses to sediments contaminated with common insecticides at concentrations below those represented by traditional toxicological endpoints (e.g., LC50). The sensitivity and ease with which this behavioral endpoint can be assayed demonstrates the potential utility of behavioral endpoints in toxicological assessments using model organisms.

Mechanistic modeling of the bioconcentration of (super)hydrophobic compounds in Hyalella azteca.

Bioconcentration tests using the freshwater amphipod Hyalella azteca as an alternative to conventional fish tests have recently received much attention. An appropriate computational model of H. azteca could help in understanding the mechanisms behind bioconcentration, in comparison to the fish as test organism. We here present the first mechanistic model for H. azteca that considers the single diffusive processes in the gills and gut. The model matches with the experimental data from the literature quite well when appropriate physiological information is used. The implementation of facilitated transport was essential for modeling. Application of the model for superhydrophobic compounds revealed binding to organic matter and the resulting decrease in bioavailable fraction as the main reason for the observed counterintuitive decrease in uptake rate constants with increasing octanol/water partition coefficient. Furthermore, estimations of the time needed to reach steady state indicated that durations of more than a month could be needed for compounds with a log Kow > 8, limiting the experimental applicability of the test. In those cases, model-based bioconcentration predictions could be a preferable approach, which could be combined with in vitro biotransformation measurements. However, our sensitivity analysis showed that the uncertainty in determining the octanol/water partition coefficients is a strong source of error for superhydrophobic compounds.

Development of insecticide resistance in Hyalella azteca.

Hyalella azteca are epibenthic amphipods that have developed resistance to pyrethroid and organophosphate insecticides due to single amino acid substitutions in the voltage-gated sodium channel and the acetylcholinesterase-1 gene, respectively. Aquatic systems are often contaminated with several different types of insecticides, therefore there is a possibility that H. azteca have also developed resistance to other classes of insecticides. The aims of the current study were to verify that pyrethroid- and organophosphate-resistant H. azteca have retained their resistance after being cultured in the absence of selective pressure for 5 years (Escondido Creek population) and 9 years (Mosher Slough population), to determine if these populations have cross-resistance to carbaryl (carbamate) and 4,4'-dichlorodiphenyltrichloroethane (DDT; organochlorine), and determine whether previous field exposure to fipronil (phenylpyrazole) and imidacloprid (neonicotinoid) caused resistance in cultured pyrethroid- and organophosphate-resistant H. azteca populations. Escondido Creek and Mosher Slough H. azteca populations both maintained high tolerances for bifenthrin due to L925I and I936F amino acid substitutions. Resistance was also found for chlorpyrifos in the Escondido Creek and Mosher Slough populations with lower genotype frequencies of the G119S substitution, indicating that additional factors may be responsible for organophosphate resistance in this study. Mosher Slough H. azteca were moderately resistant to DDT, and Escondido Creek and Mosher Slough H. azteca were moderately resistant to carbaryl, suggesting cross-resistance. No differences were observed in acute toxicity values across the three populations of H. azteca for fipronil and imidacloprid, and this is possibly due to the lack of exposure to toxic concentrations of these insecticides in the field and lack of similar modes of action to pyrethroids and organophosphates. Resistance is known to be associated with fitness costs that can place insecticide-resistant populations at risk for decline through decreased survival and reduced fecundity.

Evaluating the Functional Equivalency of Test Organism Performance in Negative and Solvent Controls During Chronic Sediment Ecotoxicity Studies Based on US Environmental Protection Agency Guidance.

The US Environmental Protection Agency (USEPA) considers sediment toxicity tests as conditional registration requirements for pesticides with soil Kd ≥50 L/kg-solid, Koc ≥1000 L/kg-organic carbon, or log Kow ≥3. The hydrophobicity of these compounds often necessitates use of solvents to ensure accurate and homogeneous dosing of spiked-sediment studies. For sediment tests, a volatile solvent (e.g., acetone) is generally used as a transient carrier. Due to low water solubility, test material is dissolved in a volatile solvent to create stock solutions. A measured aliquot of stock solution is then mixed with sand substrate, after which the solvent is evaporated. This spiking process results in negligible solvent exposure to organisms. In 2016, USEPA released final ecotoxicity test guidelines for subchronic freshwater (850.1735) and marine (850.1740) sediment test. These methods provide an option for conducting experiments with only a solvent control and no negative control. To adopt this testing strategy, functional equivalency between the negative and solvent control must be demonstrated. These test guidelines describe specific factors that should be considered for evaluating functional equivalency, including (a) the concentration of solvent in the test sediment after evaporation, (b) the levels of solvent that are known to affect organism health, (c) the known impurities in the solvent and their potential impact on organism health, and (d) the historical organism performance of solvent versus negative controls. Our analysis considers these factors and overall supports the elimination of the negative control requirement because this change is unlikely to impact the robustness or interpretability of spiked-sediment toxicity tests. Environ Toxicol Chem 2023;00:1-7. © 2023 CropLife America. Environmental Toxicology and Chemistry published by Wiley Periodicals LLC on behalf of SETAC.

Speed it up: How temperature drives toxicokinetics of organic contaminants in freshwater amphipods.

The acceleration of global climate change draws increasing attention towards interactive effects of temperature and organic contaminants. Many studies reported a higher sensitivity of aquatic invertebrates towards contaminant exposure with increasing or fluctuating temperatures. The hypothesis of this study was that the higher sensitivity of invertebrates is associated with the changes of toxicokinetic processes that determine internal concentrations of contaminants and consequently toxic effects. Therefore, the influence of temperature on toxicokinetic processes and the underlying mechanisms were studied in two key amphipod species (Gammarus pulex and Hyalella azteca). Bioconcentration experiments were carried out at four different temperatures with a mixture of 12 exposure relevant polar organic contaminants. Tissue and medium samples were taken in regular intervals and analysed by online solid-phase extraction liquid chromatography high-resolution tandem mass spectrometry. Subsequently, toxicokinetic rates were modelled and analysed in dependence of the exposure temperature using the Arrhenius equation. An exponential relationship between toxicokinetic rates versus temperature was observed and could be well depicted by applying the Arrhenius equation. Due to a similar Arrhenius temperature of uptake and elimination rates, the bioconcentration factors of the contaminants were generally constant across the temperature range. Furthermore, the Arrhenius temperature of the toxicokinetic rates and respiration was mostly similar. However, in some cases (citalopram, cyprodinil), the bioconcentration factor appeared to be temperature dependent, which could potentially be explained by the influence of temperature on active uptake mechanisms or biotransformation. The observed temperature effects on toxicokinetics may be particularly relevant in non-equilibrated systems, such as exposure peaks in summer as exemplified by the exposure modelling of a field measured pesticide peak where the internal concentrations increased by up to fourfold along the temperature gradient. The results provide novel insights into the mechanisms of chemical uptake, biotransformation and elimination in different climate scenarios and can improve environmental risk assessment.

Predicting Resistance: Quantifying the Relationship between Urban Development, Agricultural Pesticide Use, and Pesticide Resistance in a Nontarget Amphipod.

Resistance alleles within the voltage-gated sodium channel (vgsc) have been correlated with pyrethroid resistance in wild populations of the nontarget amphipod, Hyalella azteca from California (CA), U.S.A. In the present study, we expand upon the relationship between land use and the evolution of pesticide resistance in H. azteca to develop a quantitative methodology to target and screen novel populations for resistance allele genotypes in a previously uninvestigated region of the U.S. (New England: NE). By incorporating urban land development and toxicity-normalized agricultural pesticide use indices into our site selection, we successfully identified three amino acid substitutions associated with pyrethroid resistance. One of the resistance mutations has been described in H. azteca from CA (L925I). We present the remaining two (vgsc I936F and I936V) as novel pyrethroid-resistance alleles in H. azteca based on previous work in insects and elevated cyfluthrin resistance in one NE population. Our results suggest that urban pesticide use is a strong driver in the evolution of resistance alleles in H. azteca. Furthermore, our method for resistance allele screening provides an applied framework for detecting ecosystem impairment on a nationwide scale that can be incorporated into ecological risk assessment decisions.

Chronic aquatic toxicity of perfluorooctane sulfonic acid (PFOS) to Ceriodaphnia dubia, Chironomus dilutus, Danio rerio, and Hyalella azteca.

Perfluorooctane sulfonic acid (PFOS) is a ubiquitous and persistent contaminant in aquatic ecosystems. Chronic toxicity information for aquatic organisms is limited, therefore we conducted chronic PFOS toxicity tests for four model organisms commonly used for freshwater toxicology assays: Chironomus dilutus (midge), Ceriodaphnia dubia (water flea), Hyalella azteca (amphipod) and Danio rerio (zebrafish). The 16-day survival test with C. dilutus resulted in the lowest PFOS exposure concentrations to cause significant impacts, with reduced survival at 1 µg/L, a LC50 of 7.5 µg/L, and a growth EC10 of 1.5 µg/L. D. rerio was the next most sensitive species, with a 30-day LC50 of 490 µg/L and reduced growth at 260 µg/L. Effects for C. dubia and H. azteca occurred at concentrations a thousand-fold higher than for C. dilutus. H. azteca had a 42-day LC50 of 15 mg/L, an EC50 of 3.8 mg/L for reproduction (neonates per female) and an EC50 of 4.7 mg/L for growth. C. dubia was similarly tolerant of PFOS, with a 6-day LC50 of 20 mg/L for survival and an EC50 of 7 mg/L for reproduction (neonates per adult). H. azteca, C. dubia, and, to a lesser extent, D. rerio, appear tolerant of PFOS concentrations typically found in the environment. However, in agreement with previous studies, C. dilutus was particularly sensitive to PFOS exposure, with lethal and sublethal effects occurring at concentration levels present at highly contaminated sites.

Characterization of Metal Pulsed Exposure to Aquatic Organisms with Post-Exposure Monitoring: Implications for Stormwater Exposure Assessment.

Freshwater organisms are often exposed to contaminants such as heavy metals from stormwater discharges, which are dependent on rainfall duration and intensity. Therefore, standardized (48- or 96-h) continuous exposure methods developed for whole effluent toxicity (WET) testing might not always accurately convey the effects of stormwater and runoff contaminants. The present study characterized the acute toxicity of copper (Cu), zinc (Zn), and cadmium (Cd) to freshwater amphipods (Hyalella azteca) and cadmium (Cd) to water fleas (Ceriodaphnia dubia) using a modified exposure design that integrated relevant pulsed durations and included post-exposure monitoring. Less than 24-h-old C. dubia and 7 to 8-day-old H. azteca were exposed to water spiked with Cu, Zn, or Cd using 6-, 12-, 26-, or 96-h durations under standard laboratory conditions and monitored for cumulative mortality and reproduction (C. dubia only). Lethal effect (LC10s, LC25s, LC50s) and reproductive effect (EC25s, EC50s) were determined based on either mortality or reproduction of organisms at the end of each pulse (6, 12, or 26 h) and at the end of their respective tests (96 h). For all metals exposed to each organism, acute toxicity was found to be highest for the (96 h) continuous exposures. For pulsed exposures, mortality continued to increase following transfer to clean water for post-exposure monitoring. These results indicate a latent effect of Cu, Zn, and Cd to H. azteca and Cd to C. dubia. The present study concluded that using the continuous (48- or 96-h) WET exposure method overestimates the effects of stormwater and runoff contaminants. However, pulsed exposures without post-exposure monitoring also underestimate the toxicity of contaminants. The proposed pulsed exposure design provides a compromise that is more realistic than current WET methods to assess impacts from episodic events and accounts for potential latent effects that may be overlooked without monitoring post-exposure. Environ Toxicol Chem 2022;41:2488-2499. © 2022 SETAC.

Invertebrate metal accumulation and toxicity from sediments affected by the Mount Polley mine disaster.

On August 4, 2014, a tailings dam failed at the Mount Polley copper and gold mine near Likely, British Columbia, Canada, releasing approximately 25 M m[Formula: see text] of contaminated water and solid tailings material into Polley and Quesnel lakes. Water, sediment, freshwater scuds (Hyalella azteca), and mayfly larvae (Ephemeroptera) were collected during the summer of 2018 from Polley Lake, affected and unaffected sites in Quesnel Lake, and both mine-contaminated and clean far-field sites as references. Analytical results indicated that invertebrates from sites affected by the tailings breach had elevated metal concentrations relative to those from non-affected or reference sites. We conducted a controlled laboratory exposure to determine if laboratory-reared Hyalella azteca metal concentrations were related to field-collected water or sediments from the same sites as the field study. Half of the replicates prevented amphipods from directly contacting sediments (water-only exposure), while the other half allowed them direct access (sediment and water exposure). Whole-body Cu concentration was highest in Hyalella exposed to substrate from the most contaminated sites as well as in treatments where they were allowed direct access to sediments. Hyalella having direct access to metal-contaminated sediments showed reduced survival and growth relative to those in reference or control treatments. These results suggest that metals from the fine sediments associated with the Mount Polley mine disaster are bioavailable and potentially toxic to epibenthic invertebrates, even several years after the initial breach.

Pseudocorynosoma enrietti (Molfi & Freitas Fernandes, 1953) (Acanthocephala: Polymorphidae) from Patagonia (Argentina): life cycle, localities, and new host records.

All Pseudocorynosoma species inhabit freshwater environments of the American continent, but little is known about their life cycles. We report Pseudocorynosoma enrietti (Molfi & Freitas Fernandes, 1953) from natural and experimental specimens in Patagonia and identify the intermediate and definitive hosts of its life cycle for the first time in South America. Adult worms were recovered from Anas platyrhynchos (Linnaeus) and from a new definitive host, Coscoroba coscoroba Molina. Naturally infected amphipods, Hyalella patagonica Ortmann, were collected to obtain cystacanths that were fed to Gallus gallus domesticus (Linnaeus) and Anas platyrhynchos. Specimens of P. enrietti are described in detail using light and scanning electron microscopy. A key to species of the genus Pseudocorynosoma is included. Worms are characterized in both sexes by fore-trunk spines, and genital spines in an isolated field. The proboscis has 19-20 hook rows; males have 9-11 (10) hooks per row and females 7-9 (8). Males with four cement glands similar in size. Eggs elongated, with filaments. Experimental male and female worms were recovered from A. platyrhynchos at seven and 14 days, post-infection.

Effects of temperature and salinity on bioconcentration and toxicokinetics of permethrin in pyrethroid-resistant Hyalella azteca.

Recent studies demonstrated pyrethroid resistance associated with voltage-gated sodium channel mutations in populations of the epibenthic amphipod, Hyalella azteca. Resistant populations were able to tolerate and bioconcentrate pyrethroids at concentrations significantly higher than toxic levels for non-resistant populations. In conjunction with elevated bioconcentration potential, environmental alteration particularly as a result of global climate change is anticipated to significantly alter abiotic parameters including temperature and salinity. These changes are expected to influence uptake and biotransformation of contaminants. Thus, the aims of the current study were a) to examine the bioconcentration potential of permethrin in two pyrethroid-resistant clades of H. azteca and b) assess the influence of temperature and salinity changes on toxicokinetic parameters. Two pyrethroid-resistant clades of H. azteca were exposed to 14C-permethrin at three salinities (0.2, 1.0 and 6.0 practical salinity units (PSU)) and temperatures (18, 23 and 28 °C). Tests were conducted for up to 36 h and uptake, elimination and biotransformation rates were calculated. Both populations demonstrated bioconcentration factors (BCFs) between five and seven times greater than published data for non-resistant H. azteca, with significant differences between clades. Calculated BCF values were comparable to field populations of resistant H. azteca, emphasizing the potential for elevated pyrethroid bioconcentration in the natural environment and increased exposure for predators consuming pyrethroid-resistant aquatic invertebrates. Alterations to temperature and salinity had no statistically significant effect on uptake or parent compound half-life in either population, though biotransformation was elevated at higher temperatures in both populations. Salinity had a variable effect between the two populations, with lower BCF values at 1.0 PSU in clade D H. azteca and greater BCFs at 6.0 PSU in clade C H. azteca. This is the first study to demonstrate the potential for future climate scenarios to influence toxicokinetics in pyrethroid-resistant aquatic organisms.

Effects of acute arsenic exposure in two different populations of Hyalella curvispina amphipods from North Patagonia Argentina.

Arsenic (As) is a toxic metalloid present in high levels in diverse regions of Argentina. The aim of this study was to determine acute As-mediated toxicity in two different populations of autochthonous Hyalella curvispina amphipods from a reference site (LB) and an agricultural one (FO) within North Patagonia Argentina. Previously, both populations exhibited significant differences in pesticide susceptibility. Lab assays were performed to determine acute lethal concentrations, as well as some biochemical parameters. Lethal concentration (LC50) values obtained after 48 and 96 hr As exposure were not significantly different between these populations, although FO amphipods appeared slightly less susceptible. LC50-48 hr values were 3.33 and 3.92 mg/L As, while LC50-96 hr values were 1.76 and 2.14 mg/L As for LB and FO amphipods. The no observed effect concentration (NOEC) values were 0.5 mg/L As. Cholinesterase (ChE) activity was significantly diminished by As acute exposure (0.5-1.5 mg/L As), indicative of a significant neurotoxic action for this metalloid in both amphipod populations. Activities of catalase (CAT) and glutathione S-transferase (GST) and levels of reduced glutathione (GSH) were differentially altered following As exposure. CAT activity was increased after 96 hr As exposure. GST activity and GSH levels were significantly elevated followed by either a decrease or a return to control values after 96 hr treatment. However, additional studies are necessary to understand the mechanisms underlying the As-mediated oxidative effects in H. curvispina. Our findings suggest that measurement of ChE activity in H. curvispina amphipods might serve as a useful biomarker of As exposure and effect.

Evaluation of silver nanoparticle acute and chronic effects on freshwater amphipod (Hyalella azteca).

Silver nanoparticles (AgNPs) are known to cause ecotoxic effects, but there are no existing derived ambient water quality criteria (AWQC) for these nanomaterials to protect freshwater aquatic life due to insufficient toxicological data. We exposed Hyalella azteca to silver nitrate, citrate-coated AgNPs (citrate-AgNPs), and polyvinylpyrrolidone-coated AgNPs (PVP-AgNPs) in a 10-day and 28-day water-only static renewal system with clean sand as a substrate for the amphipods and compared their point estimates with the United States Environmental Protection Agency (USEPA) AWQC for silver. We observed that all treatments decreased the survival, growth, and biomass of H. azteca, and the order of toxicity was AgNO3 > citrate-AgNPs > PVP-AgNPs. The LC50s of AgNO3, citrate-AgNPs, and PVP-AgNPs were 3.0, 9.6, and 296.0 µg total Ag L-1, respectively, for the acute exposure and 2.4, 3.2, and 61.4 µg total Ag L-1, respectively, for the chronic exposure. Acute and chronic EC20s of citrate-AgNPs ranged from 0.5 to 3.5 µg total Ag L-1 while that of PVP-AgNPs ranged from 31.2 to 175 µg total Ag L-1 for growth and biomass. Both Ag+ released from AgNPs and the nanoparticles contributed to the observed toxicity. The dissolution and toxicity of AgNPs were influenced by surface coating agents, particle size, and surface charge. Most point estimates for AgNPs were above AWQC for silver (4.1 µg L-1) and the lowest concentration (0.12 µg/L) at which Ag is expected to cause chronic adverse effects to freshwater aquatic life. Our study demonstrates that the current AWQC for silver, in general, is protective of freshwater aquatic life against AgNPs tested in the present study.

Bioaccumulation potential of chlorpyrifos in resistant Hyalella azteca: Implications for evolutionary toxicology.

Given extensive use of pesticides in agriculture, there is concern for unintended consequences to non-target species. The non-target freshwater amphipod, Hyalella azteca has been found to show resistance to the organophosphate (OP) pesticide, chlorpyrifos, resulting from an amino acid substitution in acetylcholinesterase (AChE), suggesting a selective pressure of unintended pesticide exposure. Since resistant organisms can survive in contaminated habitats, there is potential for them to accumulate higher concentrations of insecticides, increasing the risk for trophic transfer. In the present study, we estimated the uptake and elimination of chlorpyrifos in non-resistant US Lab, and resistant Ulatis Creek (ULC Resistant), H. azteca populations by conducting 24-h uptake and 48-h elimination toxicokinetic experiments with 14C-chlorpyrifos. Our results indicated that non-resistant H. azteca had a larger uptake clearance coefficient (1467 mL g-1 h-1) than resistant animals (557 mL g-1 h-1). The half-life derived from the toxicokinetic models also estimated that steady state conditions were reached at 13.5 and 32.5 h for US Lab and ULC, respectively. Bioaccumulation was compared between non-resistant and resistant H. azteca by exposing animals to six different environmentally relevant concentrations for 28 h. Detection of chlorpyrifos in animal tissues indicated that resistant animals exposed to high concentrations of chlorpyrifos were capable of accumulating the insecticide up to 10-fold higher compared to non-resistant animals. Metabolite analysis from the 28-h concentration experiments showed that between 20 and 50 % parent compound was detected in H. azteca. These results imply that bioaccumulation potential can be more significant in chlorpyrifos resistant H. azteca and may be an essential factor in assessing the full impacts of toxicants on critical food webs, especially in the face of increasing pesticide and chemical runoff.

Copper transformation, speciation, and detoxification in anoxic and suboxic freshwater sediments.

The complex chemistry of copper (Cu) in freshwater sediments at low concentrations is not well understood. We evaluated the transformation processes of Cu added to freshwater sediments under suboxic and anoxic conditions. Freshwater sediments from three sources in Michigan with different characteristics (Spring Creek, River Raisin, and Maple Lake) were spiked with 30 or 60 mg kg-1 Cu and incubated under a nitrogen atmosphere. After 28-d, each treatment subset was amended with organic matter (OM) to promote anoxic conditions and evaluate its effects on Cu speciation. OM addition triggered a shift from suboxic to anoxic conditions, and sequential extractions showed that Cu accordingly shifted from acid-soluble to oxidizable fractions. Extended X-ray absorption fine-structure (EXAFS) spectroscopy revealed that Cu sulfides dominated all anoxic samples except for Spring Creek 30 mg kg-1, where Cu(I) was predominantly complexed to thiol groups of OM. Covellite and chalcopyrite (CuFeS2) were the predominant Cu species in nearly all anoxic samples, as determined by Raman spectroscopy, scanning electron microscopy, and X-ray absorption near-edge structure (XANES) spectroscopy. Copper reduction also occurred under suboxic conditions: for two of three sediments, around 80% had been reduced to Cu(I), while the remaining 20% persisted as Cu(II) complexed to OM. However, in the third coarsest (i.e., Spring Creek), around 50% of the Cu had been reduced, forming Cu(I)-OM complexes, while the remainder was Cu(II)-OM complexes. Toxicity tests showed that survival of H. azteca and D. magna were significantly lower in suboxic treatments. Anoxic sediments triggered a near-complete transformation of Cu to sulfide minerals, reducing its toxicity.

Attenuation of insecticide impact by a small wetland in a stream draining a horticultural basin in Argentina.

Horticulture has greatly increased in Argentina in recent decades mainly due to increasing greenhouse utilization and agrochemical consumption, thus representing a threat to adjacent water bodies. Riparian wetlands, however, could attenuate agrochemical contamination. The present work therefore compared insecticide concentrations in bottom sediments in addition to sediment toxicity to the amphipod Hyalella curvispina and investigated the macroinvertebrate composition upstream and downstream from a natural wetland in a small stream draining a basin undergoing intense horticultural production. The wetland surface was covered by macrophytes, mainly Thypha sp., and the insecticide concentrations measured downstream from the wetland were significantly lower, at roughly 19% of the upstream values. The growth rates of H. curvispina were significantly higher when exposed to the sediments downstream from the wetland, while the macroinvertebrate-assemblage composition was significantly different upstream and downstream: the snail Pomacea canaliculata was the dominant species upstream while the amphipod H. curvispina was dominant downstream. Pomacea canaliculata is often the dominant species in the regional streams draining agriculture and horticultural basins. Hyalella curvispina is sensitive to pesticide toxicity and is often dominant in streams draining extensive livestock basins and within a biosphere reserve. We conclude that riparian wetlands effectively attenuate horticulture contamination in pampean streams and should therefore be preserved and restored.

The contribution of detoxification pathways to pyrethroid resistance in Hyalella azteca.

Chronic exposure to pyrethroid insecticides can result in strong selective pressures on non-target species in aquatic systems and drive the evolution of resistance and population-level changes. Characterizing the underlying mechanisms of resistance is essential to better understanding the potential consequences of contaminant-driven microevolution. The current study found that multiple mechanisms enhance the overall tolerance of Hyalella azteca to the pyrethroid permethrin. In H. azteca containing mutations in the voltage-gated sodium channel (VGSC), both adaptation and acclimation played a role in mitigating the adverse effects of pyrethroid exposures. Pyrethroid resistance is primarily attributed to the heritable mutation at a single locus of the VGSC, resulting in reduced target-site sensitivity. However, additional pyrethroid tolerance was conferred through enhanced enzyme-mediated detoxification. Cytochrome P450 monooxygenases (CYP450) and general esterases (GE) significantly contributed to the detoxification of permethrin in H. azteca. Over time, VGSC mutated H. azteca retained most of their pyrethroid resistance, though there was some increased sensitivity from parent to offspring when reared in the absence of pyrethroid exposure. Permethrin median lethal concentrations (LC50s) declined from 1809 ng/L in parent (P0) individuals to 1123 ng/L in the first filial (F1) generation, and this reduction in tolerance was likely related to alterations in acclimation mechanisms, rather than changes to target-site sensitivity. Enzyme bioassays indicated decreased CYP450 and GE activity from P0 to F1, whereas the VGSC mutation was retained. The permethrin LC50s in resistant H. azteca were still two orders-of-magnitude higher than non-resistant populations indicating that the largest proportion of resistance was maintained through the inherited VGSC mutation. Thus, the noted variation in tolerance in H. azteca is likely associated with inducible traits controlling enzyme pathways. A better understanding of the mechanistic and genomic basis of acclimation is necessary to more accurately predict the ecological and evolutionary consequences of contaminant-driven change in H. azteca.