Trophodynamics of Per- and Polyfluoroalkyl Substances in the Food Web of a Large Atlantic Slope River.

Per- and polyfluoroalkyl substances (PFASs) have attracted scientific and regulatory attention due to their persistence, bioaccumulative potential, toxicity, and global distribution. We determined the accumulation and trophic transfer of 14 PFASs (5 short-chain and 9 long-chain) within the food web of the Yadkin-Pee Dee River of North Carolina and South Carolina, US. Food web components and pathways were determined by stable isotope analyses of producers, consumers, and organic matter. Analyses of water, sediment, organic matter, and aquatic biota revealed that PFASs were prevalent in all food web compartments. Biofilm, an aggregation of bacteria, fungi, algae, and protozoans and a basal resource for the aquatic food web, showed high PFAS accumulation (in 10 of 14 compounds), particularly for perfluorooctanoic acid, with the greatest mean concentration of 463.73 ng/g. The food web compartment with the most detections and greatest concentrations of PFASs was aquatic insects; all 14 PFASs were detected in individual aquatic insect samples (range of 1.0 (range of 0.57 to 2.33); it is possible that an unmeasured PFBS precursor may be accumulating in biota and metabolizing to PFBS, leading to a higher than expected TMFs for this compound. Our findings demonstrate the prevalence of PFASs in a freshwater food web with potential implications for ecological and human health.

Anglers' Views on Using Signs to Communicate Fish Consumption Advisories.

The purpose of this case study was to examine signs as a means of communicating fish consumption advisory information to English- and Spanish-speaking anglers in North Carolina. This study involved a group of stakeholders, including representatives from local and state agencies, non-governmental organizations, and academia, as well as 38 anglers in focus group discussions to learn about their knowledge and beliefs in the context of fishing in polluted waterways, including what they knew about fish consumption advisories and their perceptions of prototype signs. Across groups, participants identified two confusing elements of signs: the allowable number of servings of fish under advisory and distinct consumption recommendations for different subgroups. They recommended streamlined messaging and the use of visuals, lay terms, and locally relevant languages as ways to improve prototype signs. Additionally, participants identified the state wildlife agency as a common source of information about fish safety, more so than signs. These results suggest opportunities for improved communication of advisories, especially to the most at-risk populations.

Evaluation of Juvenile Freshwater Mussel Sensitivity to Multiple Forms of Florpyrauxifen-Benzyl.

ProcellaCOR® (active ingredient [ai], florpyrauxifen-benzyl) is an aquatic herbicide registered for use in 2018 for managing invasive and nuisance macrophyte species. Registration studies evaluating its acute toxicity revealed a favorable environmental profile; however, prior to this study, no information existed on the toxicity of florpyrauxifen-benzyl to native freshwater mussels (Family Unionidae), one of the most sensitive and imperiled faunal groups globally. We followed standard acute (96 h) toxicity test guidelines and exposed juvenile Fatmucket (Lampsilis siliquoidea) and Eastern Lampmussel (Lampsilis radiata) to the following formulations or compounds: ProcellaCOR SC and EC formulations, technical grade active ingredient (TGAI, florpyrauxifen-benzyl), and an analytical-grade sample of the weaker florpyrauxifen acid (FA). In all tests, the estimated median lethal concentrations to produce 50% mortality (LC50) were greater than the highest concentration tested of each formulation or compound. The no observable adverse effect concentrations (NOAEC, based on analytical recoveries measured at the highest concentration tested where no toxicity was observed) were TGAI = 26 µg/L, FA = 100,000 µg/L, ProcellaCOR® SC = 193 µg ai/L ProcellaCOR® EC = 585 µg ai/L and the NOAEC values for the registered commercial formulation products (ProcellaCOR® SC and ProcellaCOR® EC) were orders of magnitude greater (3.9× and 11.7×, respectively) than the maximum application rate (50 µg/L). Our results show that the herbicide formulations and compounds tested were not acutely toxic to juveniles of these two species of freshwater mussels, indicating minimal risk of short-term exposure from florpyrauxifen-benzyl applications in the environment for aquatic weed control. However, potential chronic or sublethal effects remain uncharacterized and warrant additional investigation.

Biomass of the Cyanobacterium Lyngbya wollei Alters Copper Algaecide Exposure and Risks to a Non-target Organism.

Nuisance algal infestations are increasing globally in distribution and frequency. Copper-based algaecides are routinely applied to control these infestations, though there is an ever-present concern of risks to non-target species. This research evaluated risks associated with a commonly applied chelated copper algaecide (Captain® XTR; SePRO Corporation) to a sentinel non-target species (Daphnia magna) and further assessed alteration of the exposure and toxicity when a nuisance mat-forming cyanobacterium, Lyngbya wollei, was present in exposures. Aqueous copper concentrations in treatments with algae significantly decreased within 1 h after treatment and averaged 57.5% of nominal amended Cu through the experiment duration. The 48 h LC50 values were 371 µg Cu/L with no algae present in exposures and increased significantly to 531 µg Cu/L when L. wollei was simultaneously exposed. This research provides information on the short-term fate of copper and hazard assessment by incorporating targeted binding ligands, as present in operational treatments.

T-cell bispecific antibodies in node-positive breast cancer: novel therapeutic avenue for MHC class I loss variants.

BACKGROUND: Tumor-infiltrating lymphocytes (TILs) represent a prognostic factor for survival in primary breast cancer (BC). Nonetheless, neoepitope load and TILs cytolytic activity are modest in BC, compromising the efficacy of immune-activating antibodies, which do not yet compete against immunogenic chemotherapy. PATIENTS AND METHODS: We analyzed by functional flow cytometry the immune dynamics of primary and metastatic axillary nodes [metastatic lymph nodes (mLN)] in early BC (EBC) after exposure to T-cell bispecific antibodies (TCB) bridging CD3ε and human epidermal growth factor receptor 2 (HER2) or Carcinoembryonic Antigen-Related Cell Adhesion Molecule 5 (CEACAM5), before and after chemotherapy. Human leukocyte antigen (HLA) class I loss was assessed by whole exome sequencing and immunohistochemistry. One hundred primary BC, 64 surrounding 'healthy tissue' and 24 mLN-related parameters were analyzed. RESULTS: HLA loss of heterozygosity was observed in EBC, at a clonal and subclonal level and was associated with regulatory T cells and T-cell immunoglobulin and mucin-domain-3 expression restraining the immuno-stimulatory effects of neoadjuvant chemotherapy. TCB bridging CD3ε and HER2 or CEACAM5 could bypass major histocompatibility complex (MHC) class I loss, partially rescuing T-cell functions in mLN. CONCLUSION: TCB should be developed in BC to circumvent low MHC/peptide complexes.

Contaminants in tropical island streams and their biota.

Environmental contamination is problematic for tropical islands due to their typically dense human populations and competing land and water uses. The Caribbean island of Puerto Rico (USA) has a long history of anthropogenic chemical use, and its human population density is among the highest globally, providing a model environment to study contaminant impacts on tropical island stream ecosystems. Polycyclic Aromatic Hydrocarbons, historic-use chlorinated pesticides, current-use pesticides, Polychlorinated Biphenyls (PCBs), and metals (mercury, cadmium, copper, lead, nickel, zinc, and selenium) were quantified in the habitat and biota of Puerto Rico streams and assessed in relation to land-use patterns and toxicological thresholds. Water, sediment, and native fish and shrimp species were sampled in 13 rivers spanning broad watershed land-use characteristics during 2009-2010. Contrary to expectations, freshwater stream ecosystems in Puerto Rico were not severely polluted, likely due to frequent flushing flows and reduced deposition associated with recurring flood events. Notable exceptions of contamination were nickel in sediment within three agricultural watersheds (range 123-336ppm dry weight) and organic contaminants (PCBs, organochlorine pesticides) and mercury in urban landscapes. At an urban site, PCBs in several fish species (Mountain Mullet Agonostomus monticola [range 0.019-0.030ppm wet weight] and American Eel Anguilla rostrata [0.019-0.031ppm wet weight]) may pose human health hazards, with concentrations exceeding the U.S. Environmental Protection Agency (EPA) consumption limit for 1 meal/month. American Eel at the urban site also contained dieldrin (range < detection-0.024ppm wet weight) that exceeded the EPA maximum allowable consumption limit. The Bigmouth Sleeper Gobiomorous dormitor, an important piscivorus sport fish, accumulated low levels of organic contaminants in edible muscle tissue (due to its low lipid content) and may be most suitable for human consumption island-wide; only mercury at one site (an urban location) exceeded EPA's consumption limit of 3 meals/month for this species. These results comprise the first comprehensive island-wide contaminant assessment of Puerto Rico streams and biota and provide natural resource and public health agencies here and in similar tropical islands elsewhere with information needed to guide ecosystem and fisheries conservation and management and human health risk assessment.

Windows of Susceptibility and Consequences of Early Life Exposures to 17ß-estradiol on Medaka (Oryzias latipes) Reproductive Success.

Estrogens and estrogen mimics are commonly found in surface waters and are associated with deleterious effects in fish populations. Impaired fertility and fecundity in fish following chronic exposures to estrogens and estrogen mimics during critical windows in development are well documented. However, information regarding differential reproductive effects of exposure within defined developmental stages remains sparse. In this study, reproductive capacity was assessed in Japanese medaka (Oryzias latipes) after exposure to two concentrations of 17ß-estradiol (E2ß; 2 ng/L and 50 ng/L) during four distinct stages of development: gonad development, gonad differentiation, development of secondary sex characteristics (SSC) and gametogenesis. Exposure to E2ß did not adversely impact survival, hatch success, growth, or genotypic ratios. In contrast, exposure to 50 ng/L E2ß during SSC development altered phenotypic ratios and SSC. Exposure to both E2ß treatments reduced reproductive capacity (fertility, fecundity) by 7.3-57.4% in adult medaka breeding pairs, with hindrance of SSC development resulting in the largest disruption in breeding capacity (51.6-57.4% decrease) in the high concentration. This study documents differential effects among four critical stages of development and provides insight into factors (window of exposure, exposure concentration and duration of exposure period) contributing to reproductive disruption in fish.

Copper-Based Aquatic Algaecide Adsorption and Accumulation Kinetics: Influence of Exposure Concentration and Duration for Controlling the Cyanobacterium Lyngbya wollei.

Filamentous mat-forming cyanobacteria are increasingly impairing uses of freshwater resources. To effectively manage, a better understanding of control measures is needed. Copper (Cu)-based algaecide formulations are often applied to reactively control nuisance cyanobacterial blooms. This laboratory research assessed typical field exposure scenarios for the ability of Cu to partition to, and accumulate in Lyngbya wollei. Exposure factors (Cu concentration × duration) of 4, 8, 16, 24, 32 h were tested across three aqueous Cu concentrations (1, 2, 4 ppm). Results indicated that internally accumulated copper correlated with control of L. wollei, independent of adsorbed copper. L. wollei control was determined by filament viability and chlorophyll a concentrations. Similar exposure factors elicited similar internalized copper levels and consequent responses of L. wollei. Ultimately, a "concentration-exposure-time" (CET) model was created to assist water resource managers in selecting an appropriate treatment regime for a specific in-water infestation. By assessing the exposure concentration and duration required to achieve the internal threshold of copper (i.e., critical burden) that elicits control, water management objectives can be achieved while simultaneously decreasing the environmental loading of copper and potential for non-target species risks.

Metabolomic, behavioral, and reproductive effects of the aromatase inhibitor fadrozole hydrochloride on the unionid mussel Lampsilis fasciola.

Androgen-induced masculinization of female aquatic biota poses concerns for natural population stability. This research evaluated the effects of a twelve day exposure of fadrozole hydrochloride on the metabolism and reproductive status of the unionid mussel Lampsilis fasciola. Although this compound is not considered to be widespread in the aquatic environment, it was selected as a model aromatase (enzyme that converts testosterone to estradiol) inhibitor. Adult mussels were exposed to a control and 3 concentrations of fadrozole (2µg/L, 20µg/L, and 50µg/L), and samples of gill tissue were taken on days 4 and 12 for metabolomics analysis. Gills were used because of the variety of critical processes they mediate, such as feeding, ion exchange, and siphoning. Daily observed mussel behavior included female mantle display, foot protrusion, siphoning, and larval (glochidia) releases. Glochidia mortality was significantly higher in the 20µg/L treatment. Fewer conglutinate (packets of glochidia) releases were observed in the 50µg/L treatment, and mortality was highly correlated to release numbers. Foot protrusion was significantly higher in females in nearly all treatments, including the control, during the first 4days of observations. However, this sex difference was observed only in the 50µg/L treatment during the last 8days. Generally, metabolites were significantly altered in female gill tissue in the 2µg/L treatment whereas males were mostly affected only at the highest (50µg/L) treatment. Both sexes also revealed significant reductions in fadrozole-induced metabolic effects in gill tissue sampled after 12days compared to tissue sampled after 4days, indicating time-dependent mechanisms of disruptions in metabolic pathways and homeostatic processes to compensate for such disruptions.

Rapid decreases in salinity, but not increases, lead to immune dysregulation in Nile tilapia, Oreochromis niloticus (L.).

Rapid changes in salinity, as with other environmental stressors, can have detrimental effects on fish and may trigger increased susceptibility to disease. However, the precise mechanisms of these effects are not well understood. We examined the effects of sudden increases or decreases in salinity on teleost immune function using Nile tilapia, Oreochromis niloticus (L.), as the fish model in a battery of bioassays of increasing immune system specificity. Two different salinity experiments were performed: one of increasing salinity (0 to 5, 10 and 20 g L(-1) ) and one of decreasing salinity (20 to 15, 10 and 5 g L(-1) ). Histopathology of anterior kidney, gills, gonads, intestines and liver of exposed fish was performed, but no remarkable lesions were found that were attributable to the salinity treatment regimes. The spleen was removed from each fish for analysis of cytokine expression, and peripheral blood was used for haematology, cortisol and phagocytosis assays. In the increasing salinity experiments, no significant changes were observed in any immune system assays. However, in the decreasing salinity experiments, lymphopenia, neutrophilia and monocytosis were observed in the peripheral blood without modification of the packed cell volume, plasma protein or plasma cortisol levels. Phagocytosis was increased in response to decreases in salinity from 20 g L(-1) to 15 g L(-1) , 10 g L(-1) and 5 g L(-1) , whereas phagocytic index was not significantly altered. Transforming growth factor-ß (TGF-ß) transcription increased during the same decreases in salinity. However, the TGF-ß value at 5 g L(-1) was less than those in the 15 and 10 g L(-1) salinity treatments. Interleukin-1ß (IL-1ß) transcription did not significantly respond to either salinity regime. In total, acute salinity changes appeared to trigger reactive dysregulation of the immune response in tilapia, a situation which, when combined with additional co-occurring stressors such as sudden changes in temperature and/or dissolved oxygen, could make fish more susceptible to infectious diseases. Accordingly, these findings may help to explain how sudden environmental changes may initiate disease outbreaks and lead to critical declines in cultured or wild fish populations.

Histological evaluations of organ tissues reveal sublethal effects in a freshwater mussel (Villosa iris) exposed to chloride and potassium concentrations below benchmark estimates.

Salinization of freshwater ecosystems due to anthropogenic sources will increasingly impact biodiversity. An example of point-source industrial salinization has occurred from historical activities at a U.S. Environmental Protection Agency Superfund Site near Saltville, Virginia USA and its associated chemical waste ponds adjacent to the North Fork Holston River. These point source discharges are documented contributors to mussel declines, partially due to high concentrations of chloride (Cl-, ≤ 26,000 mg Cl-/L) and potassium (K+, ≤ 97 mg K+/L). During a chronic 61-day laboratory study, Rainbow mussels, Villosa iris, were exposed to concentrations of Cl- (0, 416, 831, and 1,663 mg/L) and K+ (0, 4, 8, and 17 mg/L) to determine effects on survival and organ tissues. All test mussels died by day-2 in the 1,663 mg Cl-/L exposure, and 50% of mussels died by day-13 in the 17 mg K+/L concentration. Significantly greater abundances of tissue abnormalities were observed in digestive glands and kidneys with exposures to the 4 and 8 mg/L concentrations of K+ versus the control, and significantly greater abundances of lesions in kidneys were observed in the 416 and 831 mg Cl-/L concentrations compared to the control. The sublethal effects to digestive glands and kidneys were below reported effect (EC50, 20, 10 and LOEC) concentrations. Significant histological differences between control and baseline (day-0 sample) mussels were observed, suggesting the need for further study on the effects of captivity during longer-term laboratory experiments.

Total mercury, methylmercury, and selenium concentrations in blue marlin Makaira nigricans from a long-term dataset in the western north Atlantic.

Mercury in seafood is a neurotoxicant that threatens human health. Dynamic rates of mercury emission, re-emission, and atmospheric deposition warrant studies into mercury concentrations in fish because many are consumed by humans and can serve as sentinels of mercury levels in the environment. We modeled trends in total mercury content in an apex marine fish predator, Atlantic blue marlin Makaira nigricans, whose muscle tissues were opportunistically sampled from North Carolina (USA) sportfishing tournaments over a discontinuous time period: between 1975 and 77 and 1998-2021 (n = 148). The model-estimated influence of marlin weight on total mercury concentration was constant across years (shared slope) allowing for comparisons of weight-corrected mercury concentrations among years. Weight-corrected total mercury concentrations revealed an inter-decadal decline of approximately 45 % between the 1970s and late 1990s and then variable but relatively stable concentrations through 2021. The mean (SD) wet weight concentration of total mercury was 9.47 (4.11) from 1975 to 77 and 4.17 (2.61) from 2020 to 2021. Methylmercury and selenium were measured on a subset of fish to address questions related to human health and consumption. Methylmercury levels (mean = 0.72 µg/g) were much lower than total mercury (mean = 4.69 µg/g) indicating that total mercury is not a good proxy for methylmercury in Atlantic blue marlin. Selenium, examined as a Se:Hg molar ratio and as a selenium health benefit value (HBVSe), showed high protective value against mercury toxicity. Long-term trends in the concentration of mercury in blue marlin should continue to be monitored to determine whether policies to mitigate anthropogenic contributions to global mercury are achieving their intended goals and to provide information to inform safe human consumption.

Assessing the Toxicity of Sea Salt to Early Life Stages of Freshwater Mussels: Implications for Sea Level Rise in Coastal Rivers.

Sea levels across the planet are rising, particularly along the eastern coast of the United States. Climate-induced sea level rise can result in the inundation and intrusion of seawater into freshwater drainages. This would alter salinity regimes and lead to the salinization of coastal freshwater ecosystems. Increased salinity levels in freshwater can negatively affect freshwater-dependent species, including native mussels belonging to the order Unionida, which are highly sensitive to changes in water quality. Sea salt is largely made up of sodium and chloride ions, forming sodium chloride, a known toxicant to freshwater mussels. However, sea salt is a mixture that also contains other major ions, including potassium, sulfate, calcium, strontium, and magnesium, among others. Freshwater mussels exposed to sea salt would be exposed to each of the sea salt ions at the same time, resulting in a mixture toxicity effect. The mixture toxicity of these ions on early life stages of freshwater mussels is largely unknown because most research to date has evaluated individual salt ions in relative isolation. Therefore, we conducted acute toxicity tests on early life stages (glochidia and juvenile) of three freshwater mussel species that inhabit Atlantic Slope drainages (nonsalinity-adapted Atlanticoncha ochracea, salinity-adapted A. ochracea, Sagittunio nasutus, and Utterbackiana implicata). Glochidia and juveniles of each species were exposed to a control and six concentrations of Instant Ocean® Sea Salt (IOSS), a synthetic sea salt that closely resembles the ionic composition of natural sea salt. Exposure concentrations were 1 part(s) per thousand (ppt), 2 ppt, 8.5 ppt, 12.5 ppt, 17 ppt, and 34 ppt. We calculated the median effect concentration (EC50) for each of the eight acute toxicity tests and found that glochidia were more sensitive than juveniles to IOSS. At hour 24 EC50s for the glochidia ranged from 0.38 to 3.6 ppt, with the most sensitive freshwater mussel being the nonsalinity-adapted A. ochracea, exhibiting an EC50 of 0.38 ppt (95% confidence interval [CI] 0.33-0.44). Juvenile freshwater mussels exhibited EC50s at hour 96 ranging from 5.0 to 10.4 ppt, with the least sensitive freshwater mussel being the nonsalinity-adapted A. ochracea, exhibiting an EC50 of 10.4 ppt (95% CI 9.1-12.0). Our results show that acute exposure to sea salt adversely affects freshwater mussel viability, particularly glochidia. This information can be used to enhance freshwater mussel conservation strategies in regions that are or will be impacted by climate-induced sea level rise and associated freshwater salinization. Environ Toxicol Chem 2023;42:2478-2489. © 2023 The Authors. Environmental Toxicology and Chemistry published by Wiley Periodicals LLC on behalf of SETAC.

Review and Development of Best Practices for Toxicity Tests with Dreissenid Mussels.

Since their introduction to North America in the 1980s, research to develop effective control tools for invasive mussels (Dreissena polymorpha and D. rostriformis bugensis) has been ongoing across various research institutions using a range of testing methods. Inconsistencies in experimental methods and reporting present challenges for comparing data, repeating experiments, and applying results. The Invasive Mussel Collaborative established the Toxicity Testing Work Group (TTWG) in 2019 to identify "best practices" and guide development of a standard framework for dreissenid mussel toxicity testing protocols. We reviewed the literature related to laboratory-based dreissenid mussel toxicity tests and determined the degree to which standard guidelines have been used and their applicability to dreissenid mussel testing. We extracted detailed methodology from 99 studies from the peer-reviewed and gray literature and conducted a separate analysis for studies using presettlement and postsettlement mussels. We identified specific components of methods and approaches that could be refined or standardized for dreissenid mussels. These components included species identification, collection methods, size/age class distinction, maintenance practices, testing criteria, sample size, response measures, reporting parameters, exposure methods, and mortality criteria. We consulted experts in the field of aquatic toxicology and dreissenid mussel biology on our proposed. The final recommendations contained in the present review are based on published standard guidelines, methods reported in the published and gray literature, and the expertise of TTWG members and an external panel. In addition, our review identifies research needs for dreissenid mussel testing including improved methods for early-life stage testing, comparative data on life stages and between dreissenid mussel species, inclusion of a reference toxicant, and additional testing of nontarget species (i.e., other aquatic organisms). Environ Toxicol Chem 2023;42:1649-1666. © 2023 His Majesty the King in Right of Canada. Environmental Toxicology and Chemistry published by Wiley Periodicals LLC on behalf of SETAC. Reproduced with the permission of the Minister of Environment and Climate Change Canada. This article has been contributed to by U.S. Government employees and their work is in the public domain in the USA.

Effects of lead on Na⁺, K⁺-ATPase and hemolymph ion concentrations in the freshwater mussel Elliptio complanata.

Freshwater mussels are an imperiled fauna exposed to a variety of environmental toxicants such as lead (Pb) and studies are urgently needed to assess their health and condition to guide conservation efforts. A 28-day laboratory toxicity test with Pb and adult Eastern elliptio mussels (Elliptio complanata) was conducted to determine uptake kinetics and to assess the toxicological effects of Pb exposure. Test mussels were collected from a relatively uncontaminated reference site and exposed to a water-only control and five concentrations of Pb (as lead nitrate) ranging from 1 to 245 µg/L in a static renewal test with a water hardness of 42 mg/L. Endpoints included tissue Pb concentrations, hemolymph Pb and ion (Na⁺, K⁺, Cl⁻, Ca²âº) concentrations, and Na⁺, K⁺-ATPase enzyme activity in gill tissue. Mussels accumulated Pb rapidly, with tissue concentrations increasing at an exposure-dependent rate for the first 2 weeks, but with no significant increase from 2 to 4 weeks. Mussel tissue Pb concentrations ranged from 0.34 to 898 µg/g dry weight, were strongly related to Pb in test water at every time interval (7, 14, 21, and 28 days), and did not significantly increase after day 14. Hemolymph Pb concentration was variable, dependent on exposure concentration, and showed no appreciable change with time beyond day 7, except for mussels in the greatest exposure concentration (245 µg/L), which showed a significant reduction in Pb by 28 days, suggesting a threshold for Pb binding or elimination in hemolymph at concentrations near 1000 µg/g. The Na⁺, K⁺-ATPase activity in the gill tissue of mussels was significantly reduced by Pb on day 28 and was highly correlated with tissue Pb concentration (R² = 0.92; P = 0.013). The Na⁺, K⁺-ATPase activity was correlated with reduced hemolymph Na⁺ concentration at the greatest Pb exposure when enzyme activity was at 30% of controls. Hemolymph Ca²âº concentration increased significantly in mussels from the greatest Pb exposure and may be due to remobilization from the shell in an attempt to buffer the hemolymph against Pb uptake and toxicity. We conclude that Na⁺, K⁺-ATPase activity in mussels was adversely affected by Pb exposure, however, because the effects on activity were variable at the lower test concentrations, additional research is warranted over this range of exposures.

Survival and Contaminants in Imperiled and Common Riverine Fishes Assessed with an In Situ Bioassay Approach.

An in situ bioassay approach was used to determine whether aquatic contaminant stressors in a large Atlantic river ecosystem affect the survival of 3 fish species: the largemouth bass (Micropterus salmoides, juveniles), the fathead minnow (Pimephales promelas, adults), and the robust redhorse (Moxostoma robustum, juveniles). Hatchery-propagated fish were placed into cages to assess site-specific survival in the Yadkin-Pee Dee River of North Carolina and South Carolina, USA. Contaminants were measured in caged fish and sediment and surface water at each site. No apparent longitudinal trends in fish survival were detected, and contaminant concentrations varied among sites. Juvenile largemouth bass and robust redhorse did not survive past 13 and 23 d, with corresponding Kaplan-Meier median survival estimates of 9.7 and 12.1 d, respectively. Survival of adult fathead minnows deployed in cages alongside the juvenile fish averaged 43% at the end of the 28-d exposure, with a 22-d median survival estimate. The intersex condition, an indicator of endocrine disruption, was not observed in any adult fathead minnow. Contaminant accumulation in surviving fathead minnows was apparent, with highest accumulated concentrations of polychlorinated biphenyls (34.6-93.4 ng/g dry wt), organochlorine pesticides (19.9-66.1 ng/g dry wt), and mercury (0.17-0.63 µg/g dry wt). Contaminants and other water quality stressors in this river system appear to detrimentally impact juvenile fish survival, with presumed effects at the fish assemblage and community levels. Environ Toxicol Chem 2021;40:2206-2219. © 2021 SETAC.

Understanding the influence of multiple pollutant stressors on the decline of freshwater mussels in a biodiversity hotspot.

The Clinch River watershed of the upper Tennessee River Basin of Virginia and Tennessee, USA supports one of North America's greatest concentrations of freshwater biodiversity, including 46 extant species of native freshwater mussels (Order Unionida), 20 of which are protected as federally endangered. Despite the global biological significance of the Clinch River, mussel populations are declining in some reaches, both in species richness and abundance. The aim of this study was to evaluate the exposure of adult resident mussels to a suite of inorganic and organic contaminant stressors in distinct sections of the Clinch River that encompassed a range of mussel abundance and health. To provide insight into the potential role of pollutants in the decline of mussels, including within a previously documented "zone of mussel decline", the mainstem Clinch River (8 sites) and its tributaries (4 sites) were examined over two consecutive years. We quantified and related metals and organic contaminant concentrations in mussels to their associated habitat compartments (bed sediment, suspended particulate sediment, pore water, and surface water). We found that concentrations of organic contaminants in resident mussels, particularly the suite of 42 polycyclic aromatic hydrocarbons (PAHs) analyzed, were related to PAH concentrations in all four habitat (media) compartments. Further, PAH concentrations in mussel tissue (range 37.8-978.1 ng/g dry weight in 2012 and 194.3-1073.7 ng/g dry weight in 2013) were negatively related to the spatial pattern in mussel densities (rs = -0.64, p ≤ 0.05 in 2012 and rs = -0.83, p ≤ 0.05 in 2013) within the river, and were highest in the "zone of mussel decline". In contrast, the suite of 22 metals analyzed in resident mussels were largely unrelated to the spatial pattern of variation of metals in the four habitat compartments except for Manganese (Mn; range 3630.5-23,749.2 µg/g dry weight in 2012 and 1540.4-12,605.8 µg/g dry weight in 2013) in surface water (rs = 0.58, p < 0.1) and pore water (rs = 0.76, p ≤ 0.05). This study revealed that PAHs and Mn are important pollutant stressors to mussels in the Clinch River and that they are largely being delivered through the Guest River tributary watershed. Accordingly, future conservation and management efforts would benefit by identifying, and ideally mitigating, the sources of PAHs, Mn, and other current or legacy mining-associated pollutants to the mainstem river and its tributaries.

Method Development for a Short-Term 7-Day Toxicity Test with Unionid Mussels.

The US Environmental Protection Agency's short-term freshwater effluent test methods include a fish (Pimephales promelas), a cladoceran (Ceriodaphnia dubia), and a green alga (Raphidocelis subcapitata). There is a recognized need for additional taxa to accompany the three standard species for effluent testing. An appropriate additional taxon is unionid mussels because mussels are widely distributed, live burrowed in sediment and filter particles from the water column for food, and exhibit high sensitivity to a variety of contaminants. Multiple studies were conducted to develop a relevant and robust short-term test method for mussels. We first evaluated the comparative sensitivity of two mussel species (Villosa constricta and Lampsilis siliquoidea) and two standard species (P. promelas and C. dubia) using two mock effluents prepared by mixing ammonia and five metals (cadmium, copper, nickel, lead, and zinc) or a field-collected effluent in 7-day exposures. Both mussel species were equally or more sensitive (more than two-fold) to effluents compared with the standard species. Next, we refined the mussel test method by first determining the best feeding rate of a commercial algal mixture for three age groups (1, 2, and 3 weeks old) of L. siliquoidea in a 7-day feeding experiment, and then used the derived optimal feeding rates to assess the sensitivity of the three ages of juveniles in a 7-day reference toxicant (sodium chloride [NaCl]) test. Juvenile mussels grew substantially (30%-52% length increase) when the 1- or 2-week-old mussels were fed 2 ml twice daily and the 3-week-old mussels were fed 3 ml twice daily. The 25% inhibition concentrations (IC25s) for NaCl were similar (314-520 mg Cl/L) among the three age groups, indicating that an age range of 1- to 3-week-old mussels can be used for a 7-day test. Finally, using the refined test method, we conducted an interlaboratory study among 13 laboratories to evaluate the performance of a 7-day NaCl test with L. siliquoidea. Eleven laboratories successfully completed the test, with more than 80% control survival and reliable growth data. The IC25s ranged from 296 to 1076 mg Cl/L, with a low (34%) coefficient of variation, indicating that the proposed method for L. siliquoidea has acceptable precision. Environ Toxicol Chem 2021;40:3392-3409. © 2021 SETAC.

Thermal tolerance of juvenile freshwater mussels (Unionidae) under the added stress of copper.

Freshwater mussels fulfill an essential role in aquatic communities, but are also one of the most sensitive and rapidly declining faunal groups in North America. Rising water temperatures, caused by global climate change or industrial discharges, can further challenge impaired unionid communities, but thermal stress is almost certainly not the only stressor affecting freshwater mussels. Metals, such as copper (Cu), are a common source of toxicant exposure in aquatic environments. The toxic effects of Cu on the early life stages of freshwater mussels have been well studied, and freshwater mussels are more sensitive to Cu than most aquatic organisms. The purpose of the present study was to determine the effect of a sublethal copper concentration on the upper thermal tolerance of three species, Lampsilis siliquoidea, Potamilus alatus, and Ligumia recta, of juvenile freshwater mussels in 48- and 96-h tests. Thermal tolerance was determined over a range of experimental temperatures (20-42 degrees C) at three acclimation temperatures (17, 22, and 27 degrees C). Median lethal temperatures (LT50s) were calculated in the absence and presence of Cu, and at 48 h ranged from 34.6 to 44.4 degrees C (mean 37.7 degrees C) without Cu, and from 33.8 to 38.9 degrees C (mean 35.8 degrees C) with Cu. The LT50s at 96 h ranged from 32.5 to 35.6 degrees C (mean 34.5 degrees C) without Cu and from 33.0 to 35.4 degrees C (mean 34.2 degrees C) with Cu. Potamilus alatus had a significantly lower 48 h LT50 with Cu than without Cu at the 22 degrees C acclimation temperature; there were no other significant differences in LT50s attributed to Cu. Survival trends showed limited evidence of interactive effects between copper and temperature for all three species, suggesting the combined stress of elevated temperatures and copper exposure to freshwater mussels should be further explored.

Does proximity to coal-fired power plants influence fish tissue mercury?

Much of the mercury contamination in aquatic biota originates from coal-fired power plants, point sources that release mercury into the atmosphere. Understanding mercury dynamics is primarily important because of the toxic threat mercury poses to wildlife and humans through the consumption of contaminated fish. In this study, we quantified the relative importance of proximity to coal-fired power plants on mercury accumulation in two fish species of different trophic positions. Fish, water and sediment were collected and analyzed from 14 lakes, seven near to (<10 km) and seven far from (>30 km) coal-fired power plants. Lower tissue mercury and higher tissue selenium concentrations were measured in fish collected near power plants. Moreover, mercury accumulation in fish was driven by biotic characteristics (e.g., trophic position, total length, age), waterbody characteristics (e.g., pH, dissolved organic carbon and sulfate) and distance from power plants. Proximity to an atmospheric point-source of mercury and selenium, such as a coal-fired power plant, affects the quantities of mercury and selenium accumulated in fish tissue. Differences in accumulation are hypothesized to be driven in part by selenium-mitigated reductions in fish tissue mercury near power plants. Although reduced fish tissue mercury in systems near power plants may decrease mercury-specific risks to human consumers, these benefits are highly localized and the relatively high selenium associated with these tissues may compromise ecological health.