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Tetragnathid spiders have been used as sentinels to study the biotransport of contaminants between aquatic and terrestrial environments because a significant proportion of their diet consists of adult aquatic insects. A key knowledge gap in assessing tetragnathid spiders as sentinels is understanding the consistency of the year-to-year relationship between contaminant concentrations in spiders and sediment, water, and macroinvertebrates. We collected five years of data over a seven-year investigation at a PCB contaminated-sediment site to investigate if concentrations in spiders were consistently correlated with concentrations in sediment, water, and aquatic macroinvertebrates. Despite significant year-to-year variability in spider PCB concentrations, they were not correlated with sediment concentrations (p = 0.186). However, spider PCB concentrations were significantly, positively correlated with PCB concentrations in water (p < 0.0001, annual r 2  = 0.35-0.84) and macroinvertebrates (p < 0.0001; annual r 2  = 0.59-0.71). Analysis of covariance (ANCOVA) showed that spider PCB concentrations varied consistently with water (β = 0.63) and macroinvertebrate PCB concentrations (β = 1.023) among years. Overall, this study filled a critical knowledge gap in the utilization of tetragnathid spiders as sentinels of aquatic pollution by showing that despite year-to-year changes in PCB concentrations across environmental compartments, consistent relationships existed between spiders and water and aquatic macroinvertebrates., 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.)

Background: Restoration efforts have led to the return of anadromous fish, potential source of food for the Penobscot Indian Nation, to the previously dammed Penobscot River, Maine., Objective: U.S. Environmental Protection Agency (EPA), Penobscot Indian Nation's Department of Natural Resources (PINDNR), and Agency for Toxic Substances and Disease Registry (ATSDR), measured contaminants in six species of anadromous fish. Fish tissue concentrations were then used, along with exposure parameters, to evaluate potential human and aquatic-dependent wildlife risk., Methods: PINDNR collected, filleted, froze, and shipped fish for analysis of polychlorinated biphenyls (PCBs), polybrominated diphenyl ethers (PBDEs), dioxins/furans, and per- and polyfluoroalkyl substances (PFAS). Contaminant levels were compared to reference doses (where possible) and wildlife values (WVs)., Results: Chemical concentrations ranged from 6.37 nanogram per gram (ng/g) wet weight (ww) in American Shad roe to 100 ng/g ww in Striped Bass for total PCBs; 0.851 ng/g ww in American Shad roe to 5.92 ng/g ww in large Rainbow Smelt for total PBDEs; and 0.037 ng/g ww in American Shad roe to 0.221 ng/g ww in Striped Bass for total dioxin/furans. PFAS concentrations ranged between 0.38 ng/g ww of PFBA in Alewife to 7.86 ng/g ww of PFUnA in Sea Lamprey. Dioxin/furans and PFOS levels indicated that there are potential human health risks. The WV for mink for total PCBs (72 ng/g) was exceeded in Striped Bass and the WV for Kestrel for PBDEs (8.7 ng/g) was exceeded in large Rainbow Smelt. Mammalian wildlife consuming Blueback Herring, Striped Bass, and Sea Lamprey may be at risk based on PFOS WVs from Canada., Impact: Anadromous fish returning to the Penobscot River potentially could represent the restoration of a major component of tribal traditional diet. However, information about contaminant levels in these fish is needed to guide the tribe about consumption safety. Analysis of select species of fish and risk calculations demonstrated the need for a protective approach to consumption for both humans and wildlife. This project demonstrates that wildlife can also be impacted by contamination of fish and their risks can be as great or greater than those of humans. A One Health approach addresses this discrepancy and will lead to a healthier ecosystem., (© 2023. This is a U.S. Government work and not under copyright protection in the US; foreign copyright protection may apply.)

Levels of total mercury were measured in tissue of six species of migratory fish (alewife, American shad, blueback herring, rainbow smelt, striped bass, and sea lamprey), and in roe of American shad for two consecutive years collected from the Penobscot River or its estuary. The resultant mercury levels were compared to reference doses as established in the U.S. Environmental Protection Agency (EPA) Integrated Risk Information System and wildlife values. Mercury concentrations ranged from 4 μg/kg ww in roe to 1040 μg/kg ww in sea lamprey. Sea lamprey contained the highest amounts of mercury for both seasons of sampling. Current health advisories are set at sufficient levels to protect fishers from harmful consumption of the fish for mercury alone, except for sea lamprey. Based upon published wildlife values for mink, otter, and eagle, consumption of rainbow smelt, striped bass, or sea lamprey poses a risk to mink; striped bass and sea lamprey to otter; and sea lamprey to eagle. For future consideration, the resultant data may serve as a reference point for both human health and wildlife risk assessments for the consumption of anadromous fish. U.S. EPA works with federally recognized Tribes across the nation greatly impacted by restrictions on sustenance fishing, to develop culturally sensitive risk assessments.

For decades, we have known that chemicals affect human and wildlife behavior. Moreover, due to recent technological and computational advances, scientists are now increasingly aware that a wide variety of contaminants and other environmental stressors adversely affect organismal behavior and subsequent ecological outcomes in terrestrial and aquatic ecosystems. There is also a groundswell of concern that regulatory ecotoxicology does not adequately consider behavior, primarily due to a lack of standardized toxicity methods. This has, in turn, led to the exclusion of many behavioral ecotoxicology studies from chemical risk assessments. To improve understanding of the challenges and opportunities for behavioral ecotoxicology within regulatory toxicology/risk assessment, a unique workshop with international representatives from the fields of behavioral ecology, ecotoxicology, regulatory (eco)toxicology, neurotoxicology, test standardization, and risk assessment resulted in the formation of consensus perspectives and recommendations, which promise to serve as a roadmap to advance interfaces among the basic and translational sciences, and regulatory practices.

To better understand how the sulfate (SO 4 2- ) anion may contribute to the adverse effects associated with elevated ionic strength or salinity in freshwaters, we measured the uptake and efflux of SO 4 2- in four freshwater species: the fathead minnow ( Pimephales promelas , Teleostei: Cyprinidae), paper pondshell ( Utterbackia imbecillis , Bivalvia: Unionidae), red swamp crayfish ( Procambarus clarkii , Crustacea: Cambaridae), and two-lined mayfly ( Hexagenia bilineata , Insecta: Ephemeridae). Using δ( 34 S / 32 S) stable isotope ratios and the concentrations of S and SO 4 2- , we measured the SO 4 2- influx rate ( J in ), net flux ( J net ), and efflux rate (J out ) during a 24 h exposure period. For all four species, the means of J in for SO 4 2- were positive, and J in was significantly greater than 0 at both target SO 4 2- concentrations in the fish and mollusk and at the lower SO 4 2- concentration in the crayfish. The means of J out and J net were much more variable than those for J in , but several species by target SO 4 2- concentration combinations for J out and J net , were negative, which suggests the net excretion of SO 4 2- by the animals. The results of our experiments suggest a greater regulation of SO 4 2- in freshwater animals than has been previously reported., Competing Interests: Conflicts of Interest: The authors declare no conflict of interest.

We describe initial development of microarray-based assays for detecting 4 pyrethroid pesticides (bifenthrin, cypermethrin, esfenvalerate, and permethrin) in water. To facilitate comparison of transcriptional responses with gross apical responses, we estimated concentration-mortality curves for these pyrethroids using flow-through exposures of newly hatched Daphnia magna, Pimephales promelas adults, and 24 h posthatch P. promelas. Median lethal concentration (LC50) estimates were below most reported values, perhaps attributable to the use of flow-through exposures or of measured rather than nominal concentrations. Microarray analysis of whole P. promelas larvae and brains from exposed P. promelas adults showed that assays using either tissue type can detect these pyrethroids at concentrations below LC50 values reported for between 72 and 96% of aquatic species, depending on the pesticide. These estimates are conservative because they correspond to the lowest concentrations tested. This suggests that the simpler and less expensive whole-larval assay provides adequate sensitivity for screening contexts where acute aquatic lethality is observed, but the responsible agent is not known. Gene set analysis (GSA) highlighted several Gene Ontology (GO) terms consistent with known pyrethroid action, but the implications of other GO terms are less clear. Exploration of the sensitivity of results to changes in data processing suggests robustness of the detection assay results, but GSA results were sensitive to methodological variations. Environ Toxicol Chem 2019;38:2436-2446. Published 2019 Wiley Periodicals, Inc. on behalf of SETAC. This article is a US government work, and as such, is in the public domain in the United States of America., (Published 2019 Wiley Periodicals, Inc. on behalf of SETAC. This article is a US government work, and as such, is in the public domain in the United States of America.)

Rivers in the arid Western United States face increasing influences from anthropogenic contaminants due to population growth, urbanization, and drought. To better understand and more effectively track the impacts of these contaminants, biologically-based monitoring tools are increasingly being used to complement routine chemical monitoring. This study was initiated to assess the ability of both targeted and untargeted biologically-based monitoring tools to discriminate impacts of two adjacent wastewater treatment plants (WWTPs) on Colorado's South Platte River. A cell-based estrogen assay (in vitro, targeted) determined that water samples collected downstream of the larger of the two WWTPs displayed considerable estrogenic activity in its two separate effluent streams. Hepatic vitellogenin mRNA expression (in vivo, targeted) and NMR-based metabolomic analyses (in vivo, untargeted) from caged male fathead minnows also suggested estrogenic activity downstream of the larger WWTP, but detected significant differences in responses from its two effluent streams. The metabolomic results suggested that these differences were associated with oxidative stress levels. Finally, partial least squares regression was used to explore linkages between the metabolomics responses and the chemical contaminants that were detected at the sites. This analysis, along with univariate statistical approaches, identified significant covariance between the biological endpoints and estrone concentrations, suggesting the importance of this contaminant and recommending increased focus on its presence in the environment. These results underscore the benefits of a combined targeted and untargeted biologically-based monitoring strategy when used alongside contaminant monitoring to more effectively assess ecological impacts of exposures to complex mixtures in surface waters., (Published by Elsevier Ltd.)

The egg yolk precursor protein vitellogenin is widely used as a biomarker of estrogen exposure in male fish. However, standardized methodology is lacking and little is known regarding the reproducibility of results among laboratories using different equipment, reagents, protocols, and data analysis programs. To address this data gap we tested the reproducibility across laboratories to evaluate vitellogenin gene (vtg) expression and assessed the value of using a freely available software data analysis program. Samples collected from studies of male fathead minnows (Pimephales promelas) exposed to 17α-ethinylestradiol (EE2) and minnows exposed to processed wastewater effluent were evaluated for vtg expression in 4 laboratories. Our results indicate reasonable consistency among laboratories if the free software for expression analysis LinRegPCR is used, with 3 of 4 laboratories detecting vtg in fish exposed to 5 ng/L EE2 (n = 5). All 4 laboratories detected significantly increased vtg levels in 15 male fish exposed to wastewater effluent compared with 15 male fish held in a control stream. Finally, we were able to determine that the source of high interlaboratory variability from complementary deoxyribonucleic acid (cDNA) to quantitative polymerase chain reaction (qPCR) analyses was the expression analysis software unique to each real-time qPCR machine. We successfully eliminated the interlaboratory variability by reanalyzing raw fluorescence data with independent freeware, which yielded cycle thresholds and polymerase chain reaction (PCR) efficiencies that calculated results independently of proprietary software. Our results suggest that laboratories engaged in monitoring programs should validate their PCR protocols and analyze their gene expression data following the guidelines established in the present study for all gene expression biomarkers. Environ Toxicol Chem 2017;36:3102-3107. Published 2017 Wiley Periodicals Inc. on behalf of SETAC. This article is a US government work and, as such, is in the public domain in the United States of America., (Published 2017 Wiley Periodicals Inc. on behalf of SETAC. This article is a US government work and, as such, is in the public domain in the United States of America.)

U.S. EPA conducted a national statistical survey of fish tissue contamination at 540 river sites (representing 82 954 river km) in 2008-2009, and analyzed samples for 50 persistent organic pollutants (POPs), including 21 PCB congeners, 8 PBDE congeners, and 21 organochlorine pesticides. The survey results were used to provide national estimates of contamination for these POPs. PCBs were the most abundant, being measured in 93.5% of samples. Summed concentrations of the 21 PCB congeners had a national weighted mean of 32.7 μg/kg and a maximum concentration of 857 μg/kg, and exceeded the human health cancer screening value of 12 μg/kg in 48% of the national sampled population of river km, and in 70% of the urban sampled population. PBDEs (92.0%), chlordane (88.5%) and DDT (98.7%) were also detected frequently, although at lower concentrations. Results were examined by subpopulations of rivers, including urban or nonurban and three defined ecoregions. PCBs, PBDEs, and DDT occur at significantly higher concentrations in fish from urban rivers versus nonurban; however, the distribution varied more among the ecoregions. Wildlife screening values previously published for bird and mammalian species were converted from whole fish to fillet screening values, and used to estimate risk for wildlife through fish consumption.

Influence of waterborne butachlor (BUC), a commonly used pesticide, on morphometric, biochemical, and molecular biomarkers was evaluated in juvenile, full sibling, diploid and triploid African catfish (Clarias gariepinus). Fish were exposed for 21 days to one of three concentrations of BUC [mean measured µg/L: 22, 44 or 60]. Unexposed (control) triploids were heavier and longer and had higher visceral-somatic index (VSI) than diploids. Also, they had lighter liver weight (HSI) and showed lower transcript levels of brain gonadotropin-releasing hormone (GnRH), aromatase (cyp191b) and fushi tarazu-factor (ftz-f1), and plasma testosterone levels than diploids. Butachlor treatments had no effects, in either diploid or triploid fish, on VSI, HSI, weight or length changes, condition factor (CF), levels of plasma testosterone, 17-β estradiol (E2), cortisol, cholesterol, or mRNA levels of brain tryptophan hydroxylase (tph2), forkhead box L2 (foxl2), and 11 β-hydroxysteroid dehydrogenase type 2 (11β-hsd2). Expressions of cyp191b and ftz-f1 in triploids were upregulated by the two highest concentrations of BUC. In diploid fish, however, exposures to all BUC concentrations decreased GnRH transcription and the medium BUC concentration decreased ftz-f1 transcription. Substantial differences between ploidies in basal biomarker responses are consistent with the reported impaired reproductive axis in triploid C. gariepinus. Furthermore, the present study showed the low impact of short term exposure to BUC on reproductive axis in C. gariepinus., (Copyright © 2016 Elsevier Inc. All rights reserved.)

Omics technologies have long since promised to address a number of long standing issues related to environmental regulation. Despite considerable resource investment, there are few examples where these tools have been adopted by the regulatory community, which is in part due to a focus of most studies on discovery rather than assay development. The current work describes the initial development of an omics based assay using 48h Pimephales promelas (FHM) larvae for identifying aquatic exposures to pyrethroid pesticides. Larval FHM were exposed to seven concentrations of each of four pyrethroids (permethrin, cypermethrin, esfenvalerate and bifenthrin) in order to establish dose response curves. Then, in three separate identical experiments, FHM were exposed to a single equitoxic concentration of each pyrethroid, corresponding to 33% of the calculated LC50. All exposures were separated by weeks and all materials were either cleaned or replaced between runs in an attempt to maintain independence among exposure experiments. Gene expression classifiers were developed using the random forest algorithm for each exposure and evaluated first by cross-validation using hold out organisms from the same exposure experiment and then against test sets of each pyrethroid from separate exposure experiments. Bifenthrin exposed organisms generated the highest quality classifier, demonstrating an empirical Area Under the Curve (eAUC) of 0.97 when tested against bifenthrin exposed organisms from other exposure experiments and 0.91 against organisms exposed to any of the pyrethroids. An eAUC of 1.0 represents perfect classification with no false positives or negatives. Additionally, the bifenthrin classifier was able to successfully classify organisms from all other pyrethroid exposures at multiple concentrations, suggesting a potential utility for detecting cumulative exposures. Considerable run-to-run variability was observed both in exposure concentrations and molecular responses of exposed fish across exposure experiments. The application of a calibration step in analysis successfully corrected this, resulting in a significantly improved classifier. Classifier evaluation suggested the importance of considering a number of aspects of experimental design when developing an expression based tool for general use in ecological monitoring and risk assessment, such as the inclusion of multiple experimental runs and high replicate numbers., (Published by Elsevier B.V.)

The impacts of environmental stressors on polyploid organisms are largely unknown. This study investigated changes in morphometric, molecular, and biochemical parameters in full-sibling diploid and triploid African catfish (Clarias gariepinus) in response to chlorpyrifos (CPF) exposures. Juvenile fish were exposed to three concentrations of CPF (mean measured μg/L (SD): 9.71 (2.27), 15.7 (3.69), 31.21 (5.04)) under a static-renewal condition for 21days. Diploid control groups had higher hepatosomatic index (HSI), plasma testosterone (T), and brain GnRH and cyp19a2 expression levels than triploids. In CPF-exposed groups, changes in HSI, total weight and length were different between the diploid and triploid fish. In contrast, condition factor did not alter in any of the treatments, while visceral-somatic index (VSI) changed only in diploids. In diploid fish, exposure to CPF did not change brain 11β-hsd2, ftz-f1, foxl2, GnRH or cyp19a2 mRNA levels, while reduced tph2 transcript levels compared to the control group. In contrast, 11β-hsd2 and foxl2 expression levels were changed in triploids following CPF exposures. In diploids, plasma T levels showed a linear dose-response reduction across CPF treatments correlating with liver weight and plasma total cholesterol concentrations. In contrast, no changes in plasma cholesterol and T concentrations were observed in triploids. Plasma cortisol and 17-β estradiol (E2) showed no response to CPF exposure in either ploidy. Results of this first comparison of biomarker responses to pesticide exposure in diploid and polyploid animals showed substantial differences between diploid and triploid C. gariepinus., (Copyright © 2016 Elsevier B.V. All rights reserved.)

To assess the potential exposure of aquatic ecosystems to active pharmaceutical ingredients, the authors conducted a national-scale, probability-based statistical survey of the occurrence of these compounds in surface waters of the United States. The survey included 182 sampling sites and targeted rivers with close proximity to urban areas. The 46 analytes reported represent many classes of active pharmaceutical ingredients (APIs), including antibiotics, diuretics, antihypertensives, anticonvulsants, and antidepressants. Of the 46 analytes, 37 were detected in at least 1 sampling location. Sulfamethoxazole (an antibiotic) was the most frequently detected compound, being measured in 141 of the 182 surface waters surveyed at concentrations ranging up to 570 ng/L. Ten of the compounds were detected in 20% or more of the sampling sites. Weighted means of the analytical measurements are used with the statistical survey design and analysis to provide national estimates of the extent of contamination for these APIs in the nation's urban rivers. Published 2015 Wiley Periodicals, Inc. on behalf of SETAC. This article is a US Government work and as such, is in the public domain in the United States of America., (© 2015 SETAC.)

17α-ethinylestradiol (EE2) is a synthetic estrogen that is an active ingredient in oral contraception and hormone replacement therapy. Surveys of wastewater treatment plant effluents and surface waters throughout the world have reported EE2 concentrations in the ng/L range, and these low levels can cause significant reproductive effects in fish. This study tested the effects of three environmentally relevant EE2 concentrations: 0.47, 1.54 and 3.92 ng/L using a 21 d short-term reproductive assay to investigate the effects of EE2 on fathead minnow (Pimephales promelas) reproduction. The two highest EE2 concentrations tested in this study caused significant liver gene expression and induction of vitellogenin plasma protein in male fathead minnows. Exposure to 3.92 ng EE2/L increased the production of plasma vitellogenin in the females. Plasma estradiol concentrations were significantly reduced in females exposed to 1.54 and 3.92 ng EE2/L. All three tested concentrations significantly reduced fathead minnow egg production after a 21 d exposure to EE2. The results of this study indicate that the previously reported no observed adverse effect concentration (NOAEC) for EE2 on fathead minnow egg production (1.0 ng/L) may be too high. Because all three treatments resulted in significantly reduced egg production, the lowest observed adverse effect concentration (LOAEC) for EE2 on fathead minnow egg production is 0.47 ng EE2/L. This research estimates a NOAEC for fathead minnow reproduction at 0.24 ng EE2/L following a 21 d exposure. Additionally, induction of vitellogenin is a sensitive indicator of estrogen exposure but does not appear to be predictive of fathead minnow egg production., (Published by Elsevier Ltd.)

In this Focus article, the authors ask a seemingly simple question: Are harmful algal blooms (HABs) becoming the greatest inland water quality threat to public health and aquatic ecosystems? When HAB events require restrictions on fisheries, recreation, and drinking water uses of inland water bodies significant economic consequences result. Unfortunately, the magnitude, frequency, and duration of HABs in inland waters are poorly understood across spatiotemporal scales and differentially engaged among states, tribes, and territories. Harmful algal bloom impacts are not as predictable as those from conventional chemical contaminants, for which water quality assessment and management programs were primarily developed, because interactions among multiple natural and anthropogenic factors determine the likelihood and severity to which a HAB will occur in a specific water body. These forcing factors can also affect toxin production. Beyond site-specific water quality degradation caused directly by HABs, the presence of HAB toxins can negatively influence routine surface water quality monitoring, assessment, and management practices. Harmful algal blooms present significant challenges for achieving water quality protection and restoration goals when these toxins confound interpretation of monitoring results and environmental quality standards implementation efforts for other chemicals and stressors. Whether HABs presently represent the greatest threat to inland water quality is debatable, though in inland waters of developed countries they typically cause more severe acute impacts to environmental quality than conventional chemical contamination events. The authors identify several timely research needs. Environmental toxicology, environmental chemistry, and risk-assessment expertise must interface with ecologists, engineers, and public health practitioners to engage the complexities of HAB assessment and management, to address the forcing factors for HAB formation, and to reduce the threats posed to inland surface water quality., (© 2015 SETAC.)

Endocrine-disrupting chemicals (EDCs) such as 17α-ethynylestradiol, 17β-estradiol, estrone, and para-nonylphenol have been measured in wastewater-treatment plant effluents, surface waters, sediments, and sludge and have been shown to induce liver-specific vitellogenin (vtg) messenger RNA in male fathead minnows (Pimephales promelas). The purpose of the present study was to establish minimal concentrations of select EDCs necessary to induce transcription of vtg in 48-h static renewal exposures, as measured by quantitative real-time thermal cycle amplification. Adult males were exposed to 17α-ethynylestradiol, 17β-estradiol, estrone, and para-nonylphenol. Dose-dependent increases in vtg expression were significant with all chemicals tested. The lowest concentrations of these chemicals to induce measurable vtg expression, with significant difference from respective controls, were 17α-ethynylestradiol, 2.2 ng L(-1); para-nonylphenol, 13.9 μg L(-1); 17β-estradiol, 42.7 ng L(-1); and estrone, 46.7 ng L(-1), measured as 48-h average concentrations. The present experiments were designed to frame a commonly acceptable approach for investigators who conduct static, in-laboratory proxy environmental aquatic exposures. The present study highlights the need for investigators to report in peer-reviewed submissions the observed concentration values for minimal induction levels when measuring molecular responses to chemical exposures by means of real-time polymerase chain reaction, quantitative polymerase chain reaction, or other "omic" technologies., (Published 2015 Wiley Periodicals, Inc. on behalf of SETAC. This article is a US Government work and is in the public domain in the United States of America.)

Development of methods for assessing exposure and effects of waterborne toxicants on stream invertebrate species is important to elucidate environmentally relevant information. Current protocols for freshwater invertebrate toxicity testing almost exclusively utilize cladocerans, amphipods or chironomids rather than the more typical aquatic insect taxa found in lotic systems. Centroptilum triangulifer is a parthenogenetic mayfly occurring in depositional habitats of streams and rivers of the Eastern U.S. and Canada. C. triangulifer is an ideal stream insect for toxicity testing under field and laboratory conditions because of its short life cycle, parthenogenetic mode of reproduction, and it represents a group considered sensitive to environmental stressors. In this study, a colony of C. triangulifer was reared using a defined diet of three diatoms, Mayamaea atomus var. permitis, Nitzschia cf. pusilla, and Achnanthidium minutissimum. Percent survival (⩾80%), fecundity measurements (⩾1000 eggs) and pre-egg laying weights were used as indicators of overall colony health and fitness in our laboratory water (Lab-line) and in Moderately Hard Reconstituted Water (MHRW). Lab-line reared C. triangulifer had average survival rate of 92.69% for eleven generations and 82.99% over thirteen generations. MHRW reared C. triangulifer had an average survival rate of 80.65% for four generations and three generations of fecundities greater than 1000 eggs per individual. Pre-egg laying weight and fecundity were highly correlated and a best-fit model equation was derived to estimate egg counts for future generations. Establishment of this culturing protocol provides a more ecologically relevant species for toxicity testing and aids in further stressor identification for stream bioassessments., (Published by Elsevier Ltd.)

Criteria for establishing water quality standards that are protective for 95% of the native species are generally based upon laboratory toxicity tests. These tests utilize common model organisms that have established test methods. However, for invertebrates these species represent mostly the zooplankton community and are not inclusive of all taxa. In order to examine a potential under-representation in emerging aquatic invertebrates the US Environmental Protection Agency has cultured a parthenogenetic mayfly, Centroptilum triangulifer (Ephemeroptera: Baetidae). This study established a 48h acute and a 14-day short-term chronic testing procedure for C. triangulifer and compared its sensitivity to two model invertebrates, Ceriodaphnia dubia and Daphnia magna. Toxicity tests were conducted to determine mortality and growth effects using standard reference toxicants: NaCl, KCl and CuSO4. In 48-h acute tests, the average LC50 for the mayfly was 659mgL(-1) NaCl, 1957mgL(-1) KCl, and 11μgL(-1) CuSO4. IC25 values, using dry weight as the endpoint, were 228mgL(-1) NaCl, 356mgL(-1) KCl and 5μgL(-1) CuSO4. C. triangulifer was the most sensitive species in NaCl acute and chronic growth tests. At KCl concentrations tested, C. triangulifer was less sensitive for acute tests but was equally or more sensitive than C. dubia and D. magna for growth measurements. This study determined C. triangulifer has great potential and benefits for use in ecotoxicological studies., (Published by Elsevier Ltd.)

In 2007, approximately 420,500 cubic meters of contaminated sediment were removed from the Ashtabula River by dredging. The primary objective of the present study was to monitor contaminant exposure in fish and macroinvertebrates before, during, and after dredging. This was done by measuring tissue concentrations of polychlorinated biphenyl (PCB) and polycyclic aromatic hydrocarbon (PAH) in brown bullhead catfish (Ameriurus nebulosa) and in benthic macroinvertebrates, assessing changes in DNA damage in fish liver and blood, and scoring external and histopathological lesions and anomalies in the fish. In surficial sediment PCBs and PAHs were also quantified in conjunction with the biological sampling. The results show a significant reduction in contaminant levels in both fish and macroinvertebrates following dredging, indicating the effectiveness of the remediation in reducing exposure of biota to the primary contaminants of concern. Similarly, DNA damage levels in fish collected from the Ashtabula River significantly declined following dredging; however, a similar reduction in DNA damage over time was seen in fish collected from a reference site (Conneaut Creek), making interpretation difficult. Macroinvertebrate PCB concentrations were reflective of the sediment concentrations in the areas where Hester-Dendy samplers were deployed for macroinvertebrate collection. The present study demonstrates that these methods can be used to evaluate and assess the effectiveness of remediation techniques at contaminated sediment sites., (Published 2015 Wiley Periodicals Inc. on behalf of SETAC. This article is a US government work and, as such, is in the public domain in the United States of America.)

The US Environmental Protection Agency (USEPA) develops methods and tools for evaluating risk management strategies for sediments contaminated with polychlorinated biphenyls (PCBs), polycyclic aromatic hydrocarbons (PAHs), and other legacy pollutants. Monitored natural recovery is a risk management alternative that relies on existing physical, chemical, and biological processes to contain, destroy, and/or reduce the bioavailability or toxicity of in-place contaminants. These naturally occurring processes are monitored to ensure that management and recovery are progressing as expected. One approach frequently used to evaluate the recovery of contaminated sediments and associated biota is the assessment of contaminant tissue levels, or body burden concentrations, in top trophic level fish. In the present study, aquatic invertebrates were examined as an indicator of recent exposure to PCBs. The approach aimed to determine whether invertebrates collected using artificial substrates (i.e., Hester-Dendy samplers) could be used to discriminate among contaminated sites through the analyses of PCBs in whole homogenates of macroinvertebrates. Macroinvertebrates were sorted, preserved, and analyzed for total PCBs (t-PCBs), by summing 107 PCB congeners. Macroinvertebrate body burden concentrations showed similar trends to sediment t-PCB concentrations at the sites sampled. The results indicate that macroinvertebrates can be used to assess sediment contamination among sites that have different PCB contamination levels., (Published 2015 Wiley Periodicals Inc. on behalf of SETAC. This article is a US government work and, as such, is in the public domain in the United States of America.)

The U.S. EPA conducted a national statistical survey of fish fillet tissue with a sample size of 541 sites on boatable rivers =>5th order in 2008-2009. This is the first such study of mercury (Hg) in fish tissue from river sites focused on potential impacts to human health from fish consumption to also address wildlife impacts. Sample sites were identified as being urban or non-urban. All sample mercury concentrations were above the 3.33ugkg(-1) (ppb) quantitation limit, and an estimated 25.4% (±4.4%) of the 51663 river miles assessed exceeded the U.S. EPA 300ugkg(-1) fish-tissue based water quality criterion for mercury, representing 13144±181.8 river miles. Estimates of river miles exceeding comparable aquatic life thresholds (translated from fillet concentrations to whole fish equivalents) in avian species were similar to the number of river miles exceeding the human health threshold, whereas some mammalian species were more at risk than human from lower mercury concentrations. A comparison of means from the non-urban and urban data and among three ecoregions did not indicate a statistically significant difference in fish tissue Hg concentrations at p<0.05., (Published by Elsevier Ltd.)

Aquatic organisms are exposed to a multitude of contaminants and to fully understand the impact of multiple stressors on fish populations, we must first understand the mechanism of action for each toxicant and how the combined effects manifest at the level of the individual. 17α-ethinylestradiol (EE2) has been known to cause adverse reproductive effects including reduced fecundity and fertility, intersex and skewed sex ratios in fish by mimicking naturally produced estrogen at low concentrations. Ammonia can cause adverse reproductive and mortality effects in individual fish through effects or damage to the central nervous system. Both EE2 and ammonia are found in most municipal effluents in various concentrations. A flow-through diluter system was used to test the individual effects of these two contaminants at their respective no observable adverse effect concentration (NOAEC) as well as their combined effects on fathead minnow, (Pimephales promelas) reproduction in a mixture exposure. While neither contaminant nor their mixture altered reproduction in terms of fecundity, their mixture resulted in significant fathead minnow mortality during a 21 d exposure. This study demonstrated the need to consider mixture effects when assessing risk for toxicity testing with multiple stressors., (Published by Elsevier Ltd.)

Anthropogenic disturbances, including those from developing energy resources, can alter stream chemistry significantly by elevating total dissolved solids. Field studies have indicated that mayflies (Order Ephemeroptera) are particularly sensitive to high total dissolved solids. In the present study, the authors measured 20-d growth and survivorship of larval Neocloeon triangulifer exposed to a gradient of brine salt (mixed NaCl and CaCl2 ) concentrations. Daily growth rates were reduced significantly in all salt concentrations above the control (363 µS cm(-1) ) and larvae in treatments with specific conductance >812 µS cm(-1) were in comparatively earlier developmental stages (instars) at the end of the experiment. Survivorship declined significantly when specific conductance was >1513 µS cm(-1) and the calculated 20-d 50% lethal concentration was 2866 µS cm(-1) . The present study's results provide strong experimental evidence that elevated ion concentrations similar to those observed in developing energy resources, such as oil and gas drilling or coal mining, can adversely affect sensitive aquatic insect species., (© 2014 SETAC.)

Fathead minnows are routinely cultured for use in aquatic toxicology studies. A new mass culture system described in the present study consisted of 6 stainless steel tanks, each containing 68 fish and 20 spawning substrates. Spawning results are compared with a previous system of 22 individual glass aquaria, which contained 16 fish and 4 spawning substrates per tank. During a 19-mo period, the new system produced an average of 4105 eggs/d, compared with an average of 2465 eggs/d with the previous system. Labor and maintenance were reduced with the new system. The stainless steel tanks eliminated aquaria glass breakage, and daily water use was reduced by 45%. Analysis of reference toxicant data from fish cultured using both systems indicated no change in the sensitivity of the test animals. Analyses of 2009 egg production data determined that a 6:1 to 7:1 female to male ratio had a significantly positive impact on egg production levels and that 6-mo-old breeding stock should be introduced to the spawning tanks in mid-spring for optimal egg production during the rest of the year. Implementing a stainless steel mass culture system significantly increased efficiency of egg production; reduced turnaround delay of mature animal availability for toxicity and molecular testing; and reduced labor time, costs, and inherent safety hazards, compared with glass aquaria systems., (© 2013 SETAC.)

We measured concentrations of 56 active pharmaceutical ingredients (APIs) in effluent samples from 50 large wastewater treatment plants across the US. Hydrochlorothiazide was found in every sample. Metoprolol, atenolol, and carbamazepine were found in over 90% of the samples. Valsartan had the highest concentration (5300 ng/L), and also had the highest average concentration (1600 ng/L) across all 50 samples. Estimates of potential risks to healthy human adults were greatest for six anti-hypertensive APIs (lisinopril, hydrochlorothiazide, valsartan, atenolol, enalaprilat, and metoprolol), but nevertheless suggest risks of exposure to individual APIs as well as their mixtures are generally very low. Estimates of potential risks to aquatic life were also low for most APIs, but suggest more detailed study of potential ecological impacts from four analytes (sertraline, propranolol, desmethylsertraline, and valsartan)., (Published by Elsevier Ltd.)

Triclosan (5-chloro-2-(2,4-dichlorophenoxy)phenol) is an antimicrobial found in consumer soaps and toothpaste. It is in treated wastewater effluents at low parts-per-billion concentrations, representing a potentially chronic exposure condition for biota inhabiting receiving streams. For the present study, a naturally colonized benthos was created using flow-through indoor mesocosms; then, the benthic communities were dosed to achieve different in-stream triclosan concentrations (control, 0.1 μg/L, 0.5 μg/L, 1.0 μg/L, 5.0 μg/L, and 10 μg/L) for 56 d. Water quality parameters and endpoints from bacteria to macroinvertebrates, as well as interacting abiotic components, were measured. Effects of triclosan on specific microbial endpoints were observed at all doses, including an effect on litter decomposition dynamics at doses of 1.0 μg/L and higher. Resistance of periphytic bacteria to triclosan significantly increased at doses of 0.5 μg/L and above. By the end of dosing, the antimicrobial appeared to stimulate the stream periphyton at the 3 lowest doses, while the 2 highest doses exhibited decreased stocks of periphyton, including significantly lower bacteria cell densities and cyanobacteria abundance compared with the control. Other than an effect on benthic ostracods, the changes that occurred in the periphyton did not translate to significant change in the colonizing nematodes, the macroinvertebrate community as a whole, or other measurements of stream function. The results shed light on the role a low, chronic exposure to triclosan may play in effluent-dominated streams., (© 2013 SETAC.)

Nanoparticles (NPs) are expected to make their way into the aquatic environment where sedimentation of particles will likely occur, putting benthic organisms at particular risk. Therefore, organisms such as Hyalella azteca, an epibenthic crustacean which forages at the sediment surface, is likely to have a high potential exposure. Here we show that zinc oxide (ZnO) NPs are more toxic to H. azteca compared with the corresponding metal ion, Zn(2+). Dissolution of ZnO NPs contributes about 50% of the Zn measured in the ZnO NP suspensions, and cannot account for the toxicity of these particles to H. azteca. However, gene expression analysis is unable to distinguish between the ZnO NP exposures and zinc sulfate (ZnSO4) exposures at equitoxic concentrations. These results lead us to hypothesize that ZnO NPs provide an enhanced exposure route for Zn(2+) uptake into H. azteca, and possibly other sediment dwelling organisms. Our study supports the prediction that sediment dwelling organisms are highly susceptible to the effects of ZnO NPs and should be considered in the risk assessment of these nanomaterials.

Background: There is growing concern worldwide about the role of polluted soil and water environments in the development and dissemination of antibiotic resistance., Objective: Our aim in this study was to identify management options for reducing the spread of antibiotics and antibiotic-resistance determinants via environmental pathways, with the ultimate goal of extending the useful life span of antibiotics. We also examined incentives and disincentives for action., Methods: We focused on management options with respect to limiting agricultural sources; treatment of domestic, hospital, and industrial wastewater; and aquaculture., Discussion: We identified several options, such as nutrient management, runoff control, and infrastructure upgrades. Where appropriate, a cross-section of examples from various regions of the world is provided. The importance of monitoring and validating effectiveness of management strategies is also highlighted. Finally, we describe a case study in Sweden that illustrates the critical role of communication to engage stakeholders and promote action., Conclusions: Environmental releases of antibiotics and antibiotic-resistant bacteria can in many cases be reduced at little or no cost. Some management options are synergistic with existing policies and goals. The anticipated benefit is an extended useful life span for current and future antibiotics. Although risk reductions are often difficult to quantify, the severity of accelerating worldwide morbidity and mortality rates associated with antibiotic resistance strongly indicate the need for action.

Concentrated animal feeding operation (CAFO) manure is a cost-effective fertilizer. In the Midwest, networks of subsurface tile-drains expedite transport of animal hormones and nutrients from land-applied CAFO manure to adjacent waterways. The objective of this study was to evaluate impacts of land-applied CAFO manure on fish populations and communities. Water chemistry including hormone, pesticide, and nutrient concentrations was characterized from study sites along with fish assemblage structure, growth, and endocrine disruption assessed in selected fish species. Although most CAFO water samples had hormone concentrations <1 ng/L, equivalent concentrations for 17β-E2 and 17α-TB peaked at >30 ng/L each during the period of spawning, hatching, and development for resident fishes. CAFO sites had lower fish species richness, and fishes exhibited faster somatic growth and lower reproductive condition compared to individuals from the reference site. Fathead minnows (Pimephales promelas) exposed to CAFO ditchwater during early developmental stages exhibited significantly skewed sex ratios toward males. Maximum observed hormone concentrations were well above the lowest observable effect concentrations for these hormones; however, complexities at the field scale make it difficult to directly relate hormone concentration and impacts on fish. Complicating factors include the consistent presence of pesticides and nutrients, and the difference in temperature and stream architecture of the CAFO-impacted ditches compared to the reference site (e.g., channelization, bottom substrate, shallow pools, and riparian cover).

Elutriate bioassays are among numerous methods that exist for assessing the potential toxicity of sediments in aquatic systems. In this study, interlaboratory results were compared from 96-h Ceriodaphnia dubia and Pimephales promelas static-renewal acute toxicity tests conducted independently by two laboratories using elutriate samples prepared from the same sediment. The goal of the study was to determine if the results from the elutriate tests were comparable between two U.S. Environmental Protection Agency (USEPA) laboratories when different elutriate preparation procedures were employed by each lab. Complete agreement in site characterization was attained in 22 of the 25 samples for both bioassays amongst each lab. Of the 25 samples analyzed, 10 were found to be toxic to at least one of the species tested by either laboratory. The C. dubia elutriate tests conducted by the National Exposure Research Laboratory (NERL) indicated that 7 of the 25 sediment samples were toxic, while 8 sediment samples were characterized as such in testing conducted by USEPA Region 6 (Region 6). The P. promelas elutriate tests conducted by NERL determined 8 samples as toxic, while Region 6 tests displayed toxicity in 5 of the samples. McNemar's test of symmetry for C. dubia (S = 0.33, p = 0.5637) and P. promelas (S = 3.0, p = 0.0833) tests indicated no significant differences in designating a site toxic between NERL and Region 6 laboratories. Likewise, Cohen's kappa test revealed significant agreement between NERL and Region 6 C. dubia (K = 0.7148, p < 0.01) and P. promelas (K = 0.6939, p < 0.01) elutriate tests. The authors conclude that differences in interlaboratory elutriate preparation procedures have no bearing on the ability of either the C. dubia or P. promelas bioassay testing methods to detect toxicity while yielding similar results.

Urbanization has been linked to increased concentrations of polycyclic aromatic hydrocarbons in natural waterways. This study was designed to examine the impact of urbanization and a wastewater treatment plant by investigating the impact on field-collected bluegill (Lepomis macrochirus). Results show a significant increase in DNA strand breaks in blood cells (comet assay) linked to urbanization and a reduction in DNA strand breaks downstream of the WWTP, likely the result of dilution. A laboratory study exposing L. macrochirus to the known mutagen, methyl methanesulfonate, was performed to validate the comet assay endpoints in this species. Results of the laboratory study showed that the comet assay endpoints of tail length and tail extent moment responded in a dose- and time-dependent manner. Habitat quality assessments, along with chemical concentrations of polycyclic hydrocarbons in sediments showed that habitat quality between all sites were similar and that hydrocarbons likely contributed to the DNA strand breaks observed., (Copyright © 2012 Elsevier Inc. All rights reserved.)

Background: Over the past 10-15 years, a substantial amount of work has been done by the scientific, regulatory, and business communities to elucidate the effects and risks of pharmaceuticals and personal care products (PPCPs) in the environment., Objective: This review was undertaken to identify key outstanding issues regarding the effects of PPCPs on human and ecological health in order to ensure that future resources will be focused on the most important areas., Data Sources: To better understand and manage the risks of PPCPs in the environment, we used the "key question" approach to identify the principle issues that need to be addressed. Initially, questions were solicited from academic, government, and business communities around the world. A list of 101 questions was then discussed at an international expert workshop, and a top-20 list was developed. Following the workshop, workshop attendees ranked the 20 questions by importance., Data Synthesis: The top 20 priority questions fell into seven categories: a) prioritization of substances for assessment, b) pathways of exposure, c) bioavailability and uptake, d) effects characterization, e) risk and relative risk, f ) antibiotic resistance, and g) risk management., Conclusions: A large body of information is now available on PPCPs in the environment. This exercise prioritized the most critical questions to aid in development of future research programs on the topic.

Applications for silver nanomaterials in consumer products are rapidly expanding, creating an urgent need for toxicological examination of the exposure potential and ecological effects of silver nanoparticles (AgNPs). The integration of genomic techniques into environmental toxicology has presented new avenues to develop exposure biomarkers and investigate the mode of toxicity of novel chemicals. In the present study we used a 15k oligonucleotide microarray for Daphnia magna, a freshwater crustacean and common indicator species for toxicity, to differentiate between particle specific and ionic silver toxicity and to develop exposure biomarkers for citrate-coated and PVP-coated AgNPs. Gene expression profiles revealed that AgNO(3) and AgNPs have distinct expression profiles suggesting different modes of toxicity. Major biological processes disrupted by the AgNPs include protein metabolism and signal transduction. In contrast, AgNO(3) caused a downregulation of developmental processes, particularly in sensory development. Metal responsive and DNA damage repair genes were induced by the PVP AgNPs, but not the other treatments. In addition, two specific biomarkers were developed for the environmental detection of PVP AgNPs; although further verification under different environmental conditions is needed.

Ammonia can cause adverse reproductive and mortality effects in individual fish by interacting with the central nervous system. The last published study that assessed the effects of ammonia on fathead minnow reproduction was a lifecycle study conducted in 1986. Our study's main goal was to re-evaluate ammonia toxicity on fathead minnow Pimephales promelas reproduction using a 20-day fecundity flow-through diluter method. Flow-through diluter systems have been used by regulatory agencies, such as the U.S. Environmental Protection Agency, in the past as an effective way to estimate acceptable levels of contaminants. There was a significant difference in cumulative egg production among treatments (ANOVA; F=10.167, p≤0.01, df=3). All three concentrations of ammonia tested in this study significantly reduced fecundity after 20days of exposure (Dunnett's, p≤0.05 for each treatment). The lowest un-ionized ammonia concentration (0.06mg/L at a pH of 7.3 and temperature of 25.1°C) tested during this study resulted in a 29% decrease in cumulative fecundity. Because all tested ammonia concentrations caused an effect on P. promelas reproduction, the no effect concentration was estimated to be 0.025mg/L un-ionized ammonia (2.19mg/L total ammonia-nitrogen). This estimate was determined using the U.S. EPA Toxicity Relationship Analysis Program to calculate the 10% effect concentration of ammonia on P. promelas reproduction. This value is much lower than the previous reported no effect concentration on P. promelas reproduction (0.37mg/L un-ionized ammonia or 6.43mg/L total ammonia-nitrogen) as determined from the 1986 study, which was used to determine the ammonia water quality criteria by the U.S. Environmental Protection Agency. Our results should be considered in the next revision of water quality criteria., (Published by Elsevier B.V.)

Significant amounts of antibiotics enter the environment via point and nonpoint sources. We examined the temporal dynamics of tetracycline exposure to stream periphyton and associated organisms across a logarithmically dosed-series of experimental mesocosms, designed to mimic natural conditions. Target in-stream tetracycline exposures were based on environmentally relevant concentrations in aquatic ecosystems throughout the United States (<1-100 μg L(-1)). Significant changes in the stream biotic community were observed within 7 days with in-stream tetracycline concentrations as low as 0.5 μg L(-1), including significant changes in antibiotic resistance, bacteria abundance and productivity, algae biomass, cyanobacteria, organic biomass, and nematodes. These effects were magnified with increased exposure time and dosing concentration. Recovery of the periphyton community after 28 days of exposure was dependent upon the tetracycline dose. At the highest doses, 10 and 100 μg L(-1), bacteria productivity recovered; however, bacteria, algae, and nematode abundance did not recover at the same rate and remained low even after a 28-day recovery period (of nondosing). This study demonstrates that tetracycline exposure under near-natural conditions and at concentrations currently observed in aquatic environments may have important consequences for the structure and function of stream periphyton and, potentially, public health via increasing resistance of naturally occurring bacteria.

Background: Endocrine disrupting chemicals (e.g., estrogens, androgens and their mimics) are known to affect reproduction in fish. 17α-ethynylestradiol is a synthetic estrogen used in birth control pills. 17β-trenbolone is a relatively stable metabolite of trenbolone acetate, a synthetic androgen used as a growth promoter in livestock. Both 17α-ethynylestradiol and 17β-trenbolone have been found in the aquatic environment and affect fish reproduction. In this study, we developed a physiologically-based computational model for female fathead minnows (FHM, Pimephales promelas), a small fish species used in ecotoxicology, to simulate how estrogens (i.e., 17α-ethynylestradiol) or androgens (i.e., 17β-trenbolone) affect reproductive endpoints such as plasma concentrations of steroid hormones (e.g., 17β-estradiol and testosterone) and vitellogenin (a precursor to egg yolk proteins)., Results: Using Markov Chain Monte Carlo simulations, the model was calibrated with data from unexposed, 17α-ethynylestradiol-exposed, and 17β-trenbolone-exposed FHMs. Four Markov chains were simulated, and the chains for each calibrated model parameter (26 in total) converged within 20,000 iterations. With the converged parameter values, we evaluated the model's predictive ability by simulating a variety of independent experimental data. The model predictions agreed with the experimental data well., Conclusions: The physiologically-based computational model represents the hypothalamic-pituitary-gonadal axis in adult female FHM robustly. The model is useful to estimate how estrogens (e.g., 17α-ethynylestradiol) or androgens (e.g., 17β-trenbolone) affect plasma concentrations of 17β-estradiol, testosterone and vitellogenin, which are important determinants of fecundity in fish.

The crustacean Thamnocephalus platyurus was exposed to aqueous suspensions of fullerenes C(60) and C(70) . Aqueous fullerene suspensions were formed by stirring C(60) and C(70) as received from a commercial vendor in deionized water (termed aqu/C(60) and aqu/C(70) ) for approximately 100 d. The Z-average (mean hydrodynamic) diameters of aqu/C(60) and aqu/C(70) aggregates as measured by dynamic light scattering were 517 ± 21 nm and 656 ± 39 nm (mean ± 95% confidence limit), respectively. Exposure of T. platyurus to fullerene suspensions resulted in the formation of dark masses in the digestive track visible under a stereo microscope (×40 magnification). Fullerene ingestion over 1 h of exposure was quantitatively determined after extraction and analysis by high-performance liquid chromatography-mass spectrometry (HPLC-MS). One-hour exposures (at 3 mg/L and 6 mg/L) resulted in aqu/C(60) burdens of 2.7 ± 0.4 µg/mg and 6.8 ± 1.5 µg/mg wet weight, respectively. Thin-section transmission electron microscopy (TEM) images of aqu/C(60) -exposed T. platyurus showed the formation in the gut of fullerene agglomerates (5-10 µm) that were an order of magnitude larger than the suspended fullerene agglomerates. Upon excretion, the observed fullerene agglomerates were in the 10- to 70-µm size range and settled to the bottom of the incubation wells. In contrast to the control polystyrene microspheres, which dispersed after depuration, the aqu/C(60) agglomerates (greater than two orders of magnitude larger than the suspended fullerenes) remained agglomerated for up to six months. When exposed to fullerenes, T. platyurus shows the potential to influence agglomerate size and may facilitate movement of these nanoparticles from the water column into sediment., (Copyright © 2011 SETAC.)

Anaerobic biochemical reactors (BCRs) are useful for removing metals from mining-impacted water at remote sites. Removal processes include sorption and precipitation of metal sulfides, carbonates, and hydroxides. A question of interest is whether BCRs remove aquatic toxicity. Influent and effluent samples from the Luttrell Repository and Peerless Jenny King, both in Montana, USA; Park City, Utah, USA; and Standard Mine, Colorado, USA, were examined and compared for removal of metals and aquatic toxicity. Effluent samples from Standard Mine included those having solely BCR treatment and those having BCR treatment followed by aeration in a polishing cell. Metal removal for all sites was >90%. All influent samples were acutely toxic to Ceriodaphnia dubia and Pimephales promelas; toxicity was removed following treatment, except in the Luttrell Repository and Standard Mine BCR samples. Laboratory aeration of undiluted samples eliminated (Standard Mine BCR) or significantly reduced (Luttrell Repository, 65% survival) acute toxicity, most likely through removal of hydrogen sulfide. A toxicity identification evaluation suggested that metals also might be contributing to toxicity in the Luttrell Repository effluent samples; metals other than Mn were either not detected or very low (Fe and Pb) in the Standard Mine BCR samples. Field-aerated samples were not acutely toxic, and only the Luttrell Repository and Standard Mine samples showed short-term subchronic toxicity. Overall, results indicated BCR treatment had high metal removal efficiency and that inclusion of in-field aeration was beneficial in removal of acute and short-term subchronic toxicity., (© 2010 SETAC.)

Pesticides are nearly ubiquitous in surface waters of the United States, where they often are found as mixtures. The molecular mechanisms underlying the toxic effects of sub-lethal exposure to pesticides as both individual and mixtures are unclear. The current work aims to identify and compare differentially expressed proteins in brains of male fathead minnows (Pimephales promelas) exposed for 72 h to permethrin (7.5 μg/L), terbufos (57.5 μg/L) and a binary mixture of both. Twenty-four proteins were found to be differentially expressed among all three treatments relative to the control using an ANOVA followed by a Dunnett's post hoc test (p ≤0.05). One protein was found to be differentially expressed among all treatment groups and one protein was in common between the terbufos and the mixture group. Fifteen spots were successfully sequenced using LC-MS/MS sequencing. Proteins associated with the ubiquitin-proteasome system, glycolysis, the cytoskeleton and hypoxia were enriched. As a second objective, we attempted to establish protein expression signatures (PES) for individual permethrin and terbufos exposures. We were unable to generate a useable PES for terbufos; however, the permethrin PES was able to distinguish between control and permethrin-exposed individuals in an independent experiment with an accuracy of 87.5%. This PES also accurately classified permethrin exposed individuals when the exposure occurred as part of a mixture. The identification of proteins differentially expressed as a result of pesticide exposure represent a step forward in the understanding of mechanisms of toxicity of permethrin and terbufos. They also allow a comparison of molecular responses of the binary mixture to single exposures. The permethrin PES is the first step in establishing a method to determine exposures in real-world scenarios., (Published by Elsevier B.V.)

Zinc oxide nanoparticles (ZnO NPs) are being rapidly developed for use in consumer products, wastewater treatment, and chemotherapy providing several possible routes for ZnO NP exposure to humans and aquatic organisms. Recent studies have shown that ZnO NPs undergo rapid dissolution to Zn(2+), but the relative contribution of Zn(2+) to ZnO NP bioavailability and toxicity is not clear. We show that a fraction of the ZnO NPs in suspension dissolves, and this fraction cannot account for the toxicity of the ZnO NP suspensions to Daphnia magna. Gene expression profiling of D. magna exposed to ZnO NPs or ZnSO(4) at sublethal concentrations revealed distinct modes of toxicity. There was also little overlap in gene expression between ZnO NPs and SiO(x) NPs, suggesting specificity for the ZnO NP expression profile. ZnO NPs effected expression of genes involved in cytoskeletal transport, cellular respiration, and reproduction. A specific pattern of differential expression of three biomarker genes including a multicystatin, ferritin, and C1q containing gene were confirmed for ZnO NP exposure and provide a suite of biomarkers for identifying environmental exposure to ZnO NPs and differentiating between NP and ionic exposure.

As part of the Environmental Monitoring and Assessment Program for Great River Ecosystems (EMAP-GRE), sediment samples were collected from 447 randomly selected littoral sites along the main channels of the Ohio, Missouri, and Upper Mississippi Rivers between 2004 and 2006. Toxicity of these sediment samples was measured using a 7-day Hyalella azteca survival and growth test. Sixty-five sites (14.5%) exhibited lethal toxicity, and 130 sites (29.1%) exhibited decreased growth. In the EMAP-GRE probabilistic sampling design, each sampled site had a weight associated with it that determined the length (and proportion) of the river represented by that sample point in the population. Weighted whole-river estimates indicated that of the 4721 river km sampled, sediment from 15.9 ± 3.0% of the river (752 ± 50 km) were lethally toxic, 27.4 ± 3.5% (1289 ± 57 km) were toxic by way of growth inhibition, and 40.0 ± 3.7% (1887 ± 68 km) exhibited either lethal or growth toxicity. Selected toxic samples were analyzed for 21 pesticides, 20 polychlorinated biphenyl congeners, and 6 polybrominated diphenyl ether congeners. For all of the samples tested, the concentration levels of these analytes were mostly lower than known toxicity thresholds, and neither unionized ammonia concentration nor osmotic stress (as measured by conductivity) could account for the toxicity found in sediments. The spatial pattern of sediment toxicity cannot be readily explained by urbanization or agricultural land use at the subcatchment scale. We speculate that the distribution of toxic sediment is more likely due to a combination of localized sources, including polluted tributaries, and the redistribution of contaminated sediments from upriver. The sediment toxicity results from this study will be used, in combination with other sediment, biologic, and habitat metrics and indicators collected in the EMAP-GRE study, to help interpret and assess the condition of the Ohio, Upper Mississippi, and Missouri Rivers.

Relatively little is known about the behavior and toxicity of nanoparticles in the environment. Objectives of work presented here include establishing the toxicity of a variety of silver nanoparticles (AgNPs) to Daphnia magna neonates, assessing the applicability of a commonly used bioassay for testing AgNPs, and determining the advantages and disadvantages of multiple characterization techniques for AgNPs in simple aquatic systems. Daphnia magna were exposed to a silver nitrate solution and AgNPs suspensions including commercially available AgNPs (uncoated and coated), and laboratory-synthesized AgNPs (coated with coffee or citrate). The nanoparticle suspensions were analyzed for silver concentration (microwave acid digestions), size (dynamic light scattering and electron microscopy), shape (electron microscopy), surface charge (zeta potentiometer), and chemical speciation (X-ray absorption spectroscopy, X-ray diffraction). Toxicities of filtered (100 nm) versus unfiltered suspensions were compared. Additionally, effects from addition of food were examined. Stock suspensions were prepared by adding AgNPs to moderately hard reconstituted water, which were then diluted and used straight or after filtration with 100-nm filters. All nanoparticle exposure suspensions, at every time interval, were digested via microwave digester and analyzed by inductively coupled argon plasma-optical emission spectroscopy or graphite furnace-atomic absorption spectroscopy. Dose-response curves were generated and median lethal concentration (LC50) values calculated. The LC50 values for the unfiltered particles were (in µg/L): 1.1 ± 0.1-AgNO(3) ; 1.0 ± 0.1-coffee coated; 1.1 ± 0.2-citrate coated; 16.7 ± 2.4 Sigma Aldrich Ag-nanoparticles (SA) uncoated; 31.5 ± 8.1 SA coated. LC50 values for the filtered particles were (in µg/L): 0.7 ± 0.1-AgNO(3) ; 1.4 ± 0.1-SA uncoated; 4.4 ± 1.4-SA coated. The LC50 resulting from the addition of food was 176.4 ± 25.5-SA coated. Recommendations presented in this study include AgNP handling methods, effects from sample preparation, and advantages/disadvantages of different nanoparticle characterization techniques., (Copyright © 2010 SETAC.)

Potential exposure to active pharmaceutical ingredients (APIs) in the aquatic environment is a subject of ongoing concern. We recently published maximum likely exposure rates for several hundred human prescription pharmaceuticals commonly used in the US. These rates were estimated from nationally aggregated marketing data and wastewater production rates. The accuracy of these estimates is unclear, and it is unclear how to use the national-level estimates of exposure to predict local exposure rates. In this study we compare our previous predicted environmental concentrations (PECs), which were based on marketing data, with PECs based on regulatory data. We then use local dispensing rates for 12 APIs along with local wastewater production rates to estimate the distribution of local PECs relative to national averages, in order to identify an 'application factor' suitable for converting national-level PECs into reliable bounds for local concentrations. We compare the national-level PECs and the proposed application factor with measured environmental concentrations (MECs) published in 62 recent peer-reviewed publications. Regulatory data-based national average PECs are uniformly lower than marketing data-based national average PECs, corroborating the intended conservative nature of the marketing data-based PECs. Variability in local API usage and wastewater production rates suggest local PECs may occasionally exceed national averages by about 10-fold. Multiplying national average PECs by an 'application factor' of 10 and comparing the resulting predicted maximum local PECs to published MEC data for 83 APIs corroborates the usefulness of 10-fold adjusted national PECs as a reasonable ceiling for measured environmental concentrations., (Published by Elsevier B.V.)

We measured mercury (Hg) concentrations in whole fish from the Upper Mississippi, Missouri, and Ohio Rivers to characterize the extent and magnitude of Hg contamination and to identify environmental factors influencing Hg accumulation. Concentrations were generally lower (80% of values between 20-200 ng g(-1) wet weight) than those reported for other regions (e.g., upper Midwest and Northeast U.S.). Mercury exceeded the risk threshold for belted kingfisher (Ceryle alcyon, the most sensitive species considered) in 33-75% of river length and 1-7% of river length for humans. Concentrations were lower in the Missouri than in the Mississippi and Ohio Rivers, consistent with continental-scale patterns in atmospheric Hg deposition. Body size and trophic guild were the best predictors of Hg concentrations, which were highest in large-bodied top predators. Site geochemical and landscape properties were weakly related with fish Hg. Moreover, relationships often ran contrary to conventional wisdom, and the slopes of the relationships (positive or negative) were inconsistent among fish guilds and rivers. For example, sulfate is positively associated with fish Hg concentrations but was negatively correlated with Hg in five of six regression models of tissue concentrations. Variables such as pH, acid neutralizing capacity, and total phosphorus did not occur at levels associated with high fish Hg concentrations, partially explaining the relatively low Hg values we observed.

Numerous methods exist for assessing the potential toxicity of sediments in aquatic systems. In this study, the results from 10-day bulk sediment toxicity test methods using Hyalella azteca and Chironomus tentans were compared to results from 96-h Pimephales promelas and Ceriodaphnia dubia renewed acute toxicity tests conducted using elutriate samples prepared from the same sediments. The goal of the study was to determine if the results from the elutriate tests were comparable to those obtained from the bulk sediment tests. Of the 25 samples analyzed, 16 were found to be toxic to at least one of the species tested, in either elutriate or bulk sediment tests. The C. tentans 10-day bulk sediment test was the most sensitive, with 12 sediment samples exhibiting toxicity to this species, whereas the H. azteca bulk sediment test and C. dubia 96-h elutriate test were the least sensitive, exhibiting toxicity in only 7 of the 25 sediments tested. The P. promelas elutriate test found 8 of the 25 sediments to be toxic. Based on the total number of sites found to show toxicity, results from testing indicate 96-h elutriate tests show a level of sensitivity comparable to 10-day bulk sediment tests in assessing toxicity quantitatively. However, the methods did not always find toxicity at the same sites, suggesting that the ability of elutriate tests to predict toxicity (quantitatively) is not statistically correlated with bulk sediment methods. This would indicate that a suite of toxicity test methods would provide the most complete measure of site condition; however, in circumstances where bulk sediment testing is not feasible, elutriate tests can provide a practical and credible alternative for toxicity assessment.

Great rivers of the central United States (Upper Mississippi, Missouri, and Ohio rivers) are valuable economic and cultural resources, yet until recently their ecological condition has not been well quantified. In 2004-2005, as part of the Environmental Monitoring and Assessment Program for Great River Ecosystems (EMAP-GRE), we measured legacy organochlorines (OCs) (pesticides and polychlorinated biphenyls, PCBs) and emerging compounds (polybrominated diphenyl ethers, PBDEs) in whole fish to estimate human and wildlife exposure risks from fish consumption. PCBs, PBDEs, chlordane, dieldrin and dichlorodiphenyltrichloroethane (DDT) were detected in most samples across all rivers, and hexachlorobenzene was detected in most Ohio River samples. Concentrations were highest in the Ohio River, followed by the Mississippi and Missouri Rivers, respectively. Dieldrin and PCBs posed the greatest risk to humans. Their concentrations exceeded human screening values for cancer risk in 27-54% and 16-98% of river km, respectively. Chlordane exceeded wildlife risk values for kingfisher in 11-96% of river km. PBDE concentrations were highest in large fish in the Missouri and Ohio Rivers (mean>1000 ng g(-1) lipid), with congener 47 most prevalent. OC and PBDE concentrations were positively related to fish size, lipid content, trophic guild, and proximity to urban areas. Contamination of fishes by OCs is widespread among great rivers, although exposure risks appear to be more localized and limited in scope. As an indicator of ecological condition, fish tissue contamination contributes to the overall assessment of great river ecosystems in the U.S., (Published by Elsevier B.V.)

As part of a research effort examining system-wide responses of the hypothalamic-pituitary-gonadal (HPG) axis in fish to endocrine-active chemicals (EACs) with different modes of action, zebrafish (Danio rerio) were exposed to 25 or 100 microg/L of the aromatase inhibitor fadrozole for 24, 48, or 96 h. Global transcriptional response in brain and ovarian tissue of fish exposed to 25 microg/L of fadrozole was compared to that in control fish using a commercially available, 22,000-gene oligonucleotide microarray. Transcripts altered in brain were functionally linked to differentiation, development, DNA replication, and cell cycle. Additionally, multiple genes associated with the one-carbon pool by folate pathway (KEGG 00670) were significantly up-regulated. Transcripts altered in ovary were functionally linked to cell-cell adhesion, extracellular matrix, vasculogenesis, and development. Promoter motif analysis identified GATA-binding factor 2, Ikaros 2, alcohol dehydrogenase gene regulator 1, myoblast-determining factor, and several heat shock factors as being associated with coexpressed gene clusters that were differentially expressed following exposure to fadrozole. Based on the transcriptional changes observed, it was hypothesized that fadrozole elicits neurodegenerative stress in brain tissue and that fish cope with this stress through proliferation of radial glial cells. Additionally, it was hypothesized that changes of gene expression in the ovary of fadrozole-exposed zebrafish reflect disruption of oocyte maturation and ovulation because of impaired vitellogenesis. These hypotheses and others derived from the microarray results provide a foundation for future studies aimed at understanding responses of the HPG axis to EACs and other chemical stressors.

Knowledge of possible toxic mechanisms (or modes) of action (MOA) of chemicals can provide valuable insights as to appropriate methods for assessing exposure and effects, thereby reducing uncertainties related to extrapolation across species, endpoints and chemical structure. However, MOA-based testing seldom has been used for assessing the ecological risk of chemicals. This is in part because past regulatory mandates have focused more on adverse effects of chemicals (reductions in survival, growth or reproduction) than the pathways through which these effects are elicited. A recent departure from this involves endocrine-disrupting chemicals (EDCs), where there is a need to understand both MOA and adverse outcomes. To achieve this understanding, advances in predictive approaches are required whereby mechanistic changes caused by chemicals at the molecular level can be translated into apical responses meaningful to ecological risk assessment. In this paper we provide an overview and illustrative results from a large, integrated project that assesses the effects of EDCs on two small fish models, the fathead minnow (Pimephales promelas) and zebrafish (Danio rerio). For this work a systems-based approach is being used to delineate toxicity pathways for 12 model EDCs with different known or hypothesized toxic MOA. The studies employ a combination of state-of-the-art genomic (transcriptomic, proteomic, metabolomic), bioinformatic and modeling approaches, in conjunction with whole animal testing, to develop response linkages across biological levels of organization. This understanding forms the basis for predictive approaches for species, endpoint and chemical extrapolation. Although our project is focused specifically on EDCs in fish, we believe that the basic conceptual approach has utility for systematically assessing exposure and effects of chemicals with other MOA across a variety of biological systems.