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Integrated assessment of runoff from livestock farming operations: analytical chemistry, in vitro bioassays, and in vivo fish exposures

Animal waste from livestock farming operations can contain varying levels of natural and synthetic androgens and/or estrogens, which can contaminate surrounding waterways. In the present study, surface stream water was collected from 6 basins containing livestock farming operations. Aqueous concentrations of 12 hormones were determined via chemical analyses. Relative androgenic and estrogenic activity was measured using in vitro cell assays (MDA-kb2 and T47D-Kbluc assays, respectively). In parallel, 48-h static-renewal in vivo exposures were conducted to examine potential endocrine-disrupting effects in fathead minnows. Mature fish were exposed to surface water dilutions (0%, 25%, 50%, and 100%) and 10-ng/L of 17α-ethynylestradiol or 50-ng/L of 17β-trenbolone as positive controls. Hepatic expression of vitellogenin and estrogen receptor α mRNA, gonadal ex vivo testosterone and 17β-estradiol production, and plasma vitellogenin concentrations were examined. Potentially estrogenic and androgenic steroids were detected at low nanogram per liter concentrations. In vitro estrogenic activity was detected in all samples, whereas androgenic activity was detected in only 1 sample. In vivo exposures to the surface water had no significant dose-dependent effect on any of the biological endpoints, with the exception of increased male testosterone production in 1 exposure. The present study, which combines analytical chemistry measurements, in vitro bioassays, and in vivo fish exposures, highlights the integrated value and future use of a combination of techniques to obtain a comprehensive characterization of an environmental chemical mixture.

Arkansas, Iowa, Kentucky, New York, Wisconsin↗

Concentrations and spatial patterns of organic contaminants in tree swallow ( Tachycineta bicolor ) eggs at United States and binational Great Lakes Areas of Concern, 2010–2015

Tree swallows, Tachycineta bicolor , were sampled across the Great Lakes basin in 2010 through 2015 to provide a system-wide assessment of current exposure to organic contaminants. The results provide information identified as critical by regulators to assess the “bird or animal deformity or reproductive problems” beneficial use impairment. Eggs were collected from 69 sites across all 5 Great Lakes, including 27 Areas of Concern (AOCs), some with multiple sites, and 10 sites not listed as an AOC. Concentrations of organic contaminants in eggs were quantified and compared with background and reproductive effect thresholds. Approximately 30% of AOCs had geometric mean concentrations of total polychlorinated biphenyls (PCBs) at or below average background exposure (0.34 μg/g wet wt). Exposure to polybrominated diphenyl ethers (PBDEs) was minimal, and only 3 of 27 AOCs and 1 non-AOC had geometric mean concentrations that exceeded background for tree swallows (96 ng/g wet wt). Concentrations of both PCBs and PBDEs were 10 to 20 times below the lower limit associated with impaired hatching success. In contrast, geometric mean concentrations of polychlorinated dibenzo- p -dioxin and furan (PCDD-F) toxic equivalents (TEQs) at the Saginaw River and Bay AOC and Midland, Michigan, USA (a non-AOC site), exceeded the lower limit for hatching effects (181 pg/g PCDD-F TEQs). The rest of the sites had geometric mean concentrations of PCDD-F TEQs below background levels (87 pg/g PCDD-F TEQs). Other organic contaminants, including p,p ′-dichlorodiphenyldichloroethylene, mirex, heptachlor, and chlordane, were at or below background or adverse effect concentrations.

Great Lakes↗

Acute sensitivity of a broad range of freshwater mussels to chemicals with different modes of toxic action

Freshwater mussels, one of the most imperiled groups of animals in the world, are generally underrepresented in toxicity databases used for the development of ambient water quality criteria and other environmental guidance values. Acute 96-h toxicity tests were conducted to evaluate the sensitivity of 5 species of juvenile mussels from 2 families and 4 tribes to 10 chemicals (ammonia, metals, major ions, and organic compounds) and to screen 10 additional chemicals (mainly organic compounds) with a commonly tested mussel species, fatmucket ( Lampsilis siliquoidea ). In the multi-species study, median effect concentrations (EC50s) among the 5 species differed by a factor of ≤2 for chloride, potassium, sulfate, and zinc; a factor of ≤5 for ammonia, chromium, copper, and nickel; and factors of 6 and 12 for metolachlor and alachlor, respectively, indicating that mussels representing different families or tribes had similar sensitivity to most of the tested chemicals, regardless of modes of action. There was a strong linear relationship between EC50s for fatmucket and the other 4 mussel species across the 10 chemicals ( r 2 = 0.97, slope close to 1.0), indicating that fatmucket was similar to other mussel species; thus, this commonly tested species can be a good surrogate for protecting other mussels in acute exposures. The sensitivity of juvenile fatmucket among different populations or cultured from larvae of wild adults and captive-cultured adults was also similar in acute exposures to copper or chloride, indicating captive-cultured adult mussels can reliably be used to reproduce juveniles for toxicity testing. In compiled databases for all freshwater species, 1 or more mussel species were among the 4 most sensitive species for alachlor, ammonia, chloride, potassium, sulfate, copper, nickel, and zinc; therefore, the development of water quality criteria and other environmental guidance values for these chemicals should reflect the sensitivity of mussels. In contrast, the EC50s of fatmucket tested in the single-species study were in the high percentiles (>75th) of species sensitivity distributions for 6 of 7 organic chemicals, indicating mussels might be relatively insensitive to organic chemicals in acute exposures.

Environmental Toxicology and Chemistry↗

Acute and chronic toxicity of aluminum to a unionid mussel (Lampsilis siliquoidea) and an amphipod (Hyalella azteca) in water‐only exposures

The US Environmental Protection Agency (USEPA) is reviewing the protectiveness of the national ambient water quality criteria (WQC) for aluminum (Al) and compiling a toxicity data set to update the WQC. Freshwater mussels are one of the most imperiled groups of animals in the world, but little is known about their sensitivity to Al. The objective of the present study was to evaluate acute 96‐h and chronic 28‐d toxicity of Al to a unionid mussel ( Lampsilis siliquoidea ) and a commonly tested amphipod ( Hyalella azteca ) at a pH of 6 and water hardness of 100 mg/L as CaCO 3 . The acute 50% effect concentration (EC50) for survival of both species was >6200 μg total Al/L. The EC50 was greater than all acute values in the USEPA acute Al data set for freshwater species at a pH range of 5.0 to <6.5 and hardness normalized to 100 mg/L, indicating that the mussel and amphipod were insensitive to Al in acute exposures. The chronic 20% effect concentration (EC20) based on dry weight was 163 μg total Al/L for the mussel and 409 μg total Al/L for the amphipod. Addition of the EC20s to the USEPA chronic Al data set for pH 5.0 to <6.5 would rank the mussel ( L. siliquoidea ) as the fourth most sensitive species and the amphipod ( H. azteca ) as the fifth most sensitive species, indicating the 2 species were sensitive to Al in chronic exposures. The USEPA‐proposed acute and chronic WQC for Al would adequately protect the mussel and amphipod tested; however, inclusion of the chronic data from the present study and recalculation of the chronic criterion would likely lower the proposed chronic criterion.

Environmental Toxicology and Chemistry↗

Acute toxicity of sodium chloride and potassium chloride to a unionid mussel (Lampsilis siliquoidea) in water exposures

Freshwater mussels (order Unionoida) are one of the most imperiled groups of animals in the world. However, many ambient water quality criteria and other environmental guideline values do not include data for freshwater mussels, in part because mussel toxicity test methods are comparatively new and data may not have been available when criteria and guidelines were derived. The objectives of the present study were to evaluate the acute toxicity of sodium chloride (NaCl) and potassium chloride (KCl) to larvae (glochidia) and/or juveniles of a unionid mussel (fatmucket, Lampsilis siliquoidea ) and to determine the potential influences of water hardness (50, 100, 200, and 300 mg/L as CaCO 3 ) and other major ions (Ca, K, SO 4 , or HCO 3 ) on the acute toxicity of NaCl to the mussels. From the KCl test, the 50% effect concentration (EC50) for fatmucket glochidia was 30 mg K/L, similar to or slightly lower than the EC50s for juvenile fatmucket (37–46 mg K/L) tested previously in our laboratory. From the NaCl tests, the EC50s for glochidia increased from 441 to 1597 mg Cl/L and the EC50s for juvenile mussels increased from 911 to 3092 mg Cl/L with increasing water hardness from 50 to 300 mg/L. Increasing K from 0.4 to 1.9 mg/L, SO 4 from 13 to 40 mg/L, or HCO 3 from 44 to 200 mg/L in the 50 mg/L hardness water did not substantially change the NaCl EC50s for juvenile mussels, whereas increasing Ca from 9.9 to 42 mg/L increased the EC50s by a factor of 2. The overall results indicate that glochidia were equally or more sensitive to NaCl and KCl compared with juvenile mussels and that the increased water hardness ameliorated the acute toxicity of NaCl to glochidia and juveniles. These responses rank fatmucket among the most acutely sensitive freshwater organisms to NaCl and KCl.

Missouri↗

Temperature‐related responses of an invasive mussel and 2 unionid mussels to elevated carbon dioxide

Zebra mussels (Dreissena polymorpha) have exacerbated the decline of native freshwater mussels (Order Unionida) in North America since their arrival in the 1980s. Options for controlling invasive mussels, particularly in unionid mussel habitats, are limited. Previously, carbon dioxide (CO2) showed selective toxicity for zebra mussels, relative to unionids, when applied in cool water (12 °C). We first determined 96 h lethal concentrations of CO2 at 5 and 20 °C to zebra mussels and responses of juvenile plain pocketbook (Lampsilis cardium). Next, we compared the time to lethality for zebra mussels at 5, 12, and 20 °C during exposure to partial pressure of CO2 (PCO2) 110¬–120 atmospheres (atm; 1 atm = 101.325 kPa) and responses of juvenile plain pocketbook and fragile papershell (Leptodea fragilis). We found efficacious CO2 treatment regimens at each temperature that were minimally lethal to unionids. At 5 °C, plain pocketbook survived 96 h exposure to the highest PCO2 treatment (139 atm). At 20 °C, the 96 h LC10 (lethal concentration to 10% of animals) for plain pocketbook [173 atm PCO2, 95% confidence interval (CL) 147–198 atm] was higher than the LC99 for zebra mussels (118 atm PCO2, CL 109–127 atm). Lethal time to 99% mortality (LT99) of zebra mussels in 110 to 120 atm PCO2 ranged from 100 h at 20 °C to 300 h at 5 °C. Mean survival of juvenile unionids exceeded 85% in LT99 CO2 treatments at all temperatures. Short-term infusion of 100 to 200 atm PCO2 at a range of water temperatures could reduce biofouling by zebra mussels with limited adverse effects on unionid mussels.

Environmental Toxicology and Chemistry↗

High frequency of lead exposure in the population of an endangered Australian top predator, the Tasmanian wedge-tailed eagle (Aquila audax fleayi)

Lead poisoning, mainly through incidental ingestion of lead ammunition in carcasses, is a threat to scavenging and predatory bird species worldwide. In Australia, shooting for animal control is widespread, and a range of native scavenging species are susceptible to lead exposure. However, the prevalence of lead exposure in Australia's scavenging and predatory birds is largely unknown. We evaluated the degree to which the Tasmanian wedge‐tailed eagle ( Aquila audax fleayi ), an endangered Australian raptor and facultative scavenger, showed evidence of lead exposure. We detected lead in 100% of femur and liver tissues of 109 eagle carcasses opportunistically collected throughout Tasmania between 1996 and 2018. Concentrations were elevated in 10% of 106 liver (> 6 mg/kg dw) and 4% of 108 femur (> 10 mg/kg dw) samples. We also detected lead in 96% of blood samples taken from 24 live nestlings, with 8% at elevated concentrations (> 10 μg/dL). Of the liver samples with elevated lead, 73% had lead 207/206 isotope ratios within the published range of lead‐based bullets available in Tasmania. These first comprehensive data on lead exposure of an Australian raptor are comparable to those for raptor studies elsewhere that identify lead‐based ammunition exposure as a conservation threat. Our findings highlight the importance of further research and efforts to address lead contamination throughout the Tasmanian ecosystem and in other Australian regions.

Environmental Toxicology and Chemistry↗

The sensitivity of a unionid mussel (Lampsilis siliquoidea) to a permitted effluent and elevated potassium in the effluent

Freshwater mussels are one of the most imperiled groups of animals in the world and are among the most sensitive species to a variety of chemicals. However, little is known about the sensitivity of freshwater mussels to wastewater effluents. The objectives of the present study were to (1) assess the toxicity of a permitted effluent, which entered the Deep Fork River, Oklahoma (USA), to a unionid mussel ( Lampsilis siliquoidea ) and to two standard test species (cladoceran Ceriodaphnia dubia ; and fathead minnow Pimephales promelas ) in short-term 7-day effluent tests; (2) evaluate the relative sensitivities of the three species to potassium (K), an elevated major ion in the effluent, using 7-day toxicity tests with KCl spiked into a Deep Fork River upstream reference water; (3) determine the potential influences of background water characteristics on the acute K toxicity to the mussel (96-h exposures) and cladoceran (48-h exposure) in four reconstituted waters that mimicked the hardness and ionic composition ranges of the Deep Fork River; and (4) determine the potential influence of temperature on acute K toxicity to the mussel. The effluent was found to be toxic to mussels and cladocerans, and it contained elevated concentrations of major cations and anions relative to the upstream Deep Fork River reference water. The K concentration in the effluent was 48-fold greater than in the upstream water. Compared with the standard species, the mussel was more than 4-fold more sensitive to the effluent in the 7-day effluent tests and more than 8-fold more sensitive to K in the 7-day K toxicity tests. The acute K toxicity to the mussel decreased by a factor of 2 when the water hardness was increased from soft (42 mg/L as CaCO 3 ) to very hard (314 mg/L as CaCO 3 ), whereas the acute K toxicity to the cladoceran remained almost the same as hardness increased from 84 to 307 mg/L as CaCO 3 . Acute K toxicity to the mussel at 23 °C was similar to the toxicity at an elevated temperature of 28 °C. The overall results indicate that the two standard test species may not represent the sensitivity of the tested mussel to both the effluent and K, and the toxicity of K was influenced by the hardness in test waters, but by a limited magnitude.

Oklahoma↗

Site- and individual-level contaminations affect infection prevalence of an emerging infectious disease of amphibians

Emerging infectious disease outbreaks are one of multiple stressors responsible for amphibian declines globally. In the northeastern United States, ranaviral diseases are prevalent in amphibians and other ectothermic species, but there is still uncertainty as to whether their presence is leading to population level effects. Further, there is also uncertainty surrounding the potential interactions among disease infection prevalence in free-ranging animals and habitat degradation (co-occurrence of chemical stressors). The current study was designed to provide field-based estimates of the relationship between amphibian disease and chemical stressors. We visited 40 wetlands across three protected areas, estimated the prevalence of ranavirus among populations of larval wood frogs and spotted salamanders, and assessed chemical and biological stressors in wetland habitats and larval amphibians using a suite of selected bioassays, screening tools and chemical analyses. Estimated ranavirus occupancy varied among the three protected areas and ranged from 0.27 to 0.55 with considerable variation within each protected area. Of the stressors evaluated, ranavirus prevalence was strongly and positively related to concentrations of metalloestrogens (metals with the potential to bind to estrogen receptors) and total metals in wetland sediments and weakly and negatively related to total pesticide concentrations in larval amphibians. These results can be used by land managers to refine habitat assessments to include such environmental factors with the potential to influence disease susceptibility.

Maryland, Massachusetts↗

Comparative toxicity of aquatic per- and polyfluoroalkyl substance exposure in three species of amphibians

Per- and polyfluoroalkyl substances (PFAS) are contaminants of concern due to their widespread occurrence in the environment, persistence, and potential to elicit a range of negative health effects. Per- and polyfluoroalkyl substances are regularly detected in surface waters, but their effects on many aquatic organisms are still poorly understood. Species with thyroid-dependent development, like amphibians, can be especially susceptible to PFAS effects on thyroid hormone regulation. We examined sublethal effects of aquatic exposure to four commonly detected PFAS on larval northern leopard frogs ( Rana [Lithobates] pipiens ), American toads ( Anaxyrus americanus ), and eastern tiger salamanders ( Ambystoma tigrinum ). Animals were exposed for 30 days (frogs and salamanders) or until metamorphosis (toads) to 10, 100, or 1000 μg/L of perfluorooctane sulfonate (PFOS), perfluorooctanoic acid (PFOA), perfluorohexane sulfonate (PFHxS), or 6:2 fluorotelomer sulfonate (6:2 FTS). We determined that chronic exposure to common PFAS can negatively affect amphibian body condition and development at concentrations as low as 10 µg/L. These effects were highly species dependent, with species having prolonged larval development (frogs and salamanders) being more sensitive to PFAS than more rapidly developing species (toads). Our results demonstrate that some species could experience sublethal effects at sites with surface waters highly affected by PFAS. Our results also indicate that evaluating PFAS toxicity using a single species may not be sufficient for accurate amphibian risk assessment. Future studies are needed to determine whether these differences in susceptibility can be predicted from species' life histories and whether more commonly occurring environmental levels of PFAS could affect amphibians.

Environmental Toxicology and Chemistry↗

Do pharmaceuticals in the environment pose a risk to wildlife?

The vast majority of knowledge related to the question of, “To what extent do pharmaceuticals in the environment pose a risk to wildlife?”, stems from the Asian vulture crisis (>99% decline of some species of old-world vultures on the Indian subcontinent related to the veterinary use of the non-steroidal anti-inflammatory drug (NSAID) diclofenac). The hazard of diclofenac and other NSAIDs (carprofen, flunixin, ketoprofen, nimesulide, phenylbutazone) to vultures and other avian species has since been demonstrated; indeed only meloxicam and tolfenamic acid have been found to be vulture-safe. Since diclofenac was approved for veterinary use in Spain and Italy in 2013 (home to ~95% of vultures in Europe), the risk of NSAIDs to vultures in these countries has become one of the principal concerns related to pharmaceuticals and wildlife. Many of the other bodies of work on pharmaceutical exposure, hazard and risk to wildlife also relate to adverse effects in birds, (e.g., poisoning of scavenging birds in North America and Europe from animal carcasses containing pentobarbital; secondary and even tertiary poisoning of birds exposed to pesticides used in veterinary medicine as cattle dips; migratory birds as a vector for the transfer of antimicrobial and antifungal resistance). While there is some research related to endocrine disruption in reptiles and potential exposure of aerial insectivores, there remain numerous knowledge gaps for risk posed by pharmaceuticals to amphibians, reptiles and mammals. Developing non-invasive sampling techniques and new approach methodologies (e.g., genomic, in vitro , in silico , in ovo ) are important if we are to bridge the current knowledge gaps without extensive vertebrate testing.

Environmental Toxicology and Chemistry↗

Toxicokinetics of PAHs in Hexagenia

The accumulation kinetics of two waterborne polycyclic aromatic hydrocarbons (PAHs), benzo[a]pyrene (BAP) and phenanthrene (PHE), were studied in the mayfly nymph ( Hexagenia limbata ). The uptake clearance decreased while the bioconcentration of BAP increased with an increase in weight of the H. limbata nymph. The relationship between uptake clearance and bioconcentration for PHE was variable, and bioconcentration was greater for the heavier animals. Two kinetic models were used to evaluate the effect of nymph weight on disposition of PAHs: (a) the amount-uptake clearance model, similar to models most frequently used in environmental toxicology; and (b) a clearance-volume model, similar to models used in clinical pharmacology. The two models gave similar predictive results but were different in a few cases. These differences in common parameter estimation probably resulted from methodologies used and high data variability rather than the models themselves, since they are mathematically equal. Some of the parameters are unique to each of the models and are defined and described. The clearance of oxygen from water is inversely and linearly related to the weight of the mayfly nymphs, but oxygen clearances were always much less than the uptake clearances of the PAHs. The high PAH uptake clearance compared to oxygen clearance implies a greater surface area or efficiency for PAH accumulation from water.

Environmental Toxicology and Chemistry↗

Acute toxicity of the herbicide bromoxynil to Daphnia magna

The acute toxicities of technical-grade bromoxynil octanoate (BO) and two commercial formulations, Buctril® and Bronate®, to < 24-h-old neonate Daphnia magna (Straus) were determined in soft, hard, and oligosaline water. In addition, effects of life stage, feeding, aging the herbicide, and exposure duration on BO toxicity to daphnids were investigated. Regardless of formulation, life stage, and water quality, BO was found to be extremely to highly toxic to daphnids in standard tests; 48-h EC50 values ranged from 41 to 161 m̈g/L. Bromoxynil octanoate was the most toxic to neonates in soft water and the least toxic in hard water. The acute toxicities of the three bromoxynil herbicides to a given age group of daphnids were similar within the same water type. Overall, neonates and 7-d-old adults were more sensitive than 14- or 15-d-old adults to each herbicide. Feeding daphnids during the toxicity test significantly decreased BO toxicity compared to not feeding them. Aging BO (as Buctril) in hard water decreased its toxicity, and the rate of deactivation was rapid, with an estimated half-life of biological activity of 13 h. Daphnids immobilized by exposures to toxic BO concentrations for ≤ 6 h recovered their mobility, whereas exposures of 18 and 24 h to BO produced toxic effects in daphnids similar to those exposed for 48 h. These results indicated that standard continuous exposure tests may not adequately predict the acute toxicity of BO to freshwater animals in the field.

Environmental Toxicology and Chemistry↗

Bioaccumulation of metals by Hyalella azteca exposed to contaminated sediments from the upper Clark Fork River, Montana

Macroinvertebrates contaminated with metals in the Clark Fork River of Montana have been demonstrated to be a potentially toxic component in the diet of trout Because sediment was the suspected source of metals to these invertebrates, bioaccumulation of As, Cd, Cu, Pb, and Zn from sediment was evaluated by exposing the amphipod Hyalella azteca for 28 d in the laboratory to samples of sediment collected from depositional areas of the Clark Fork River Benthic invertebrates collected from riffles adjacent to the depositional areas were also analyzed for metals The pattern of metal accumulation between laboratory-exposed and field-collected animals was similar, however, the concentrations of metals in laboratory exposed amphipods were often 50 to 75% less than were the concentrations of metals in the field collected invertebrates These findings indicate that sediment is a significant source of metals to invertebrates in the Clark Fork River Additional studies should be conducted to determine threshold concentrations for effects of dietary metals on fish Long-term monitoring of the river should include sampling benthic invertebrates for metal accumulation.

Environmental Toxicology and Chemistry↗

Blood changes in mallards exposed to white phosphorus

White phosphorus (P 4 ) has been extensively used by the military for various purposes, including marking artillery impacts and as an obscurant. Target practice in an Alaskan tidal marsh during the last 4 decades has deposited large amounts of P 4 particles in sediments and water, which have resulted in die-offs of several waterfowl species. Because the toxicity of P 4 in birds has not been well documented and because it is quickly excreted or metabolized in living animals, we sought to determine the effects of experimental dosing on blood characteristics in game farm mallards ( Anas platyrhynchos ). In two experiments, one employing single doses of 5.4 mg P 4 per kilogram body weight in corn oil and the other using daily repeated doses of pelletized P 4 at either 0.5 or 1.0 mg/kg, there were significant changes in aspartate aminotransferase, alanine aminotransferase, lactate dehydrogenase (LDH), inorganic P, hematocrit, and hemoglobin. Other indications of exposure included changes in uric acid, creatinine, and total protein, which were consistent with reported liver and kidney damage due to this contaminant. Changes in white blood cells included a greater frequency of thrombocytes and fewer lymphocytes in dosed birds compared to controls. A biomarker of exposure based on LDH activity and hemoglobin is proposed.

Environmental Toxicology and Chemistry↗

Effects of pre- and postnatal polychlorinated biphenyl exposure on metabolic rate and thyroid hormones of white-footed mice

Energy budgets have proven to be a valuable tool for predicting life history from physiological data in terrestrial vertebrates, yet these concepts have not been applied to the physiological effects of contaminants. Contaminants might affect energy budgets by imposing an additional metabolic cost or by reducing the overall amount of energy taken in; either process will reduce the energy available for production (i.e., growth or reproduction). This study examined whole animal energetic effects of polychlorinated biphenyl (PCB) exposure in white-footed mice ( Peromyscus leucopus ). Exposure to PCBs is known to reduce concentrations of plasma thyroid hormones, and thyroid hormones exert strong control over the rate of energy metabolism in mammals. Peromyscus leucopus that were proven breeders were fed PCBs in their food at 0, 10, and 25 ppm. Through lactation, offspring were exposed to PCB from conception and were maintained on the maternal diet to adulthood. No effects were seen on energy metabolism (O 2 consumption, measured in adulthood) or on growth, but there were large dose-dependent decreases in thyroid hormone concentrations, particularly T 4 . The apparent disparity in our data between unchanged metabolic rates and 50% reductions in T 4 concentrations can be rationalized by noting that free T 3 (the fraction not bound to plasma protein) in treated mice was not significantly different from controls and that metabolism is most strongly influenced by free T 3 . Overall, this study did not demonstrate any energetic consequences of PCB exposure in P. leucopus at dietary concentrations up to 25 ppm.

Environmental Toxicology and Chemistry↗

Assessing contaminants of emerging concern in the Great Lakes Ecosystem: A decade of method development and practical application

Assessing the ecological risk of contaminants in the field typically involves consideration of a complex mixture of compounds which may or may not be detected via instrumental analyses. Further, there are insufficient data to predict the potential biological effects of many detected compounds, leading to their being characterized as contaminants of emerging concern (CECs). Over the past several years, advances in chemistry, toxicology, and bioinformatics have resulted in a variety of concepts and tools that can enhance the pragmatic assessment of the ecological risk of CECs. The present Focus article describes a 10+- year multiagency effort supported through the U.S. Great Lakes Restoration Initiative to assess the occurrence and implications of CECs in the North American Great Lakes. State-of-the-science methods and models were used to evaluate more than 700 sites in about approximately 200 tributaries across lakes Ontario, Erie, Huron, Michigan, and Superior, sometimes on multiple occasions. Studies featured measurement of up to 500 different target analytes in different environmental matrices, coupled with evaluation of biological effects in resident species, animals from in situ and laboratory exposures, and in vitro systems. Experimental taxa included birds, fish, and a variety of invertebrates, and measured endpoints ranged from molecular to apical responses. Data were integrated and evaluated using a diversity of curated knowledgebases and models with the goal of producing actionable insights for risk assessors and managers charged with evaluating and mitigating the effects of CECs in the Great Lakes. This overview is based on research and data captured in approximately about 90 peer-reviewed journal articles and reports, including approximately about 30 appearing in a virtual issue comprised of highlighted papers published in Environmental Toxicology and Chemistry or Integrated Environmental Assessment and Management . Environ Toxicol Chem 2023;42:2506–2518. © 2023 SETAC. This article has been contributed to by U.S. Government employees and their work is in the public domain in the USA.

Environmental Toxicology and Chemistry↗

Inconsistent transcriptomic responses to hexabromocyclododecane in Japanese quail: A comparative analysis of results from four different study designs

Efforts to use transcriptomics for toxicity testing have classically relied on the assumption that chemicals consistently produce characteristic transcriptomic signatures that are reflective of their mechanism of action. However, the degree to which transcriptomic responses are conserved across different test methodologies has seldom been explored. With increasing regulatory demand for New Approach Methods (NAMs) that use alternatives to animal models and high‐content approaches such as transcriptomics, this type of comparative analysis is needed. We examined whether common genes are dysregulated in Japanese quail ( Coturnix japonica ) liver following sublethal exposure to the flame retardant hexabromocyclododecane (HBCD), when life stage and test methodologies differ. The four exposure scenarios included one NAM: Study 1—early‐life stage (ELS) exposure via a single egg injection, and three more traditional approaches; Study 2—adult exposure using a single oral gavage; Study 3—ELS exposure via maternal deposition after adults were exposed through their diet for 7 weeks; and Study 4—ELS exposure via maternal deposition and re‐exposure of nestlings through their diet for 17 weeks. The total number of differentially expressed genes (DEGs) detected in each study was variable (Study 1, 550; Study 2, 192; Study 3, 1; Study 4, 3) with only 19 DEGs shared between Studies 1 and 2. Factors contributing to this lack of concordance are discussed and include differences in dose, but also quail strain, exposure route, sampling time, and HBCD stereoisomer composition. The results provide a detailed overview of the transcriptomic responses to HBCD at different life stages and routes of exposure in a model avian species and highlight certain challenges and limits of comparing transcriptomics across different test methodologies.

Environmental Toxicology and Chemistry↗