Toxicological effects assessment for wildlife in the 21st century: Review of current methods and recommendations for a path forward.

Model species (e.g., granivorous gamebirds, waterfowl, passerines, domesticated rodents) have been used for decades in guideline laboratory tests to generate survival, growth, and reproductive data for prospective ecological risk assessments (ERAs) for birds and mammals, while officially adopted risk assessment schemes for amphibians and reptiles do not exist. There are recognized shortcomings of current in vivo methods as well as uncertainty around the extent to which species with different life histories (e.g., terrestrial amphibians, reptiles, bats) than these commonly used models are protected by existing ERA frameworks. Approaches other than validating additional animal models for testing are being developed, but the incorporation of such new approach methodologies (NAMs) into risk assessment frameworks will require robust validations against in vivo responses. This takes time, and the ability to extrapolate findings from nonanimal studies to organism- and population-level effects in terrestrial wildlife remains weak. Failure to adequately anticipate and predict hazards could have economic and potentially even legal consequences for regulators and product registrants. In order to be able to use fewer animals or replace them altogether in the long term, vertebrate use and whole organism data will be needed to provide data for NAM validation in the short term. Therefore, it is worth investing resources for potential updates to existing standard test guidelines used in the laboratory as well as addressing the need for clear guidance on the conduct of field studies. Herein, we review the potential for improving standard in vivo test methods and for advancing the use of field studies in wildlife risk assessment, as these tools will be needed in the foreseeable future. Integr Environ Assess Manag 2024;20:699-724. © 2023 His Majesty the King in Right of Canada and The Authors. Integrated Environmental Assessment and Management published by Wiley Periodicals LLC on behalf of Society of Environmental Toxicology & Chemistry (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.

Wildlife ecological risk assessment in the 21st century: Promising technologies to assess toxicological effects.

Despite advances in toxicity testing and the development of new approach methodologies (NAMs) for hazard assessment, the ecological risk assessment (ERA) framework for terrestrial wildlife (i.e., air-breathing amphibians, reptiles, birds, and mammals) has remained unchanged for decades. While survival, growth, and reproductive endpoints derived from whole-animal toxicity tests are central to hazard assessment, nonstandard measures of biological effects at multiple levels of biological organization (e.g., molecular, cellular, tissue, organ, organism, population, community, ecosystem) have the potential to enhance the relevance of prospective and retrospective wildlife ERAs. Other factors (e.g., indirect effects of contaminants on food supplies and infectious disease processes) are influenced by toxicants at individual, population, and community levels, and need to be factored into chemically based risk assessments to enhance the "eco" component of ERAs. Regulatory and logistical challenges often relegate such nonstandard endpoints and indirect effects to postregistration evaluations of pesticides and industrial chemicals and contaminated site evaluations. While NAMs are being developed, to date, their applications in ERAs focused on wildlife have been limited. No single magic tool or model will address all uncertainties in hazard assessment. Modernizing wildlife ERAs will likely entail combinations of laboratory- and field-derived data at multiple levels of biological organization, knowledge collection solutions (e.g., systematic review, adverse outcome pathway frameworks), and inferential methods that facilitate integrations and risk estimations focused on species, populations, interspecific extrapolations, and ecosystem services modeling, with less dependence on whole-animal data and simple hazard ratios. Integr Environ Assess Manag 2024;20:725-748. © 2023 His Majesty the King in Right of Canada and The Authors. Integrated Environmental Assessment and Management published by Wiley Periodicals LLC on behalf of Society of Environmental Toxicology & Chemistry (SETAC). Reproduced with the permission of the Minister of Environment and Climate Change Canada. This article has been contributed to by US Government employees and their work is in the public domain in the USA.

Evaluation of the Chronic Reproductive Toxicity of a Fluorine-Free Firefighting Foam and a Short-Chain Fluorinated Foam to Northern Bobwhite Quail (Colinus virginianus).

The development of fluorine-free firefighting foams has been proposed as a way to reduce the adverse environmental consequences of foams containing per- and poly-fluoroalkyl substances. While there are likely fewer environmental and ecological concerns with these new fluorine-free foams in terms of persistence and bioaccumulation, it is prudent to evaluate the ecotoxicity of these fluorine-free foam products given the absence of data. Oral chronic drinking water exposure studies on adult pairs of northern bobwhite quail (Colinus virginianus) were conducted with a short-chain fluorinated and a fluorine-free foam: Buckeye Platinum Plus C6 and National Foam Avio Green KHC, respectively, at three exposure concentrations (0.01%, 0.1%, and 0.25%). Adults were monitored for survival, growth, and reproductive output; and chicks were monitored for survival and growth. Growth parameters in adult quail were not affected by exposure to the Buckeye or Avio foam. However, liver lipid content was higher in adult males exposed to the Buckeye foam or the Avio foam at the highest exposure concentrations. Chicks were heavier and had higher growth rates after adult exposure to Avio at the highest exposure level (0.25%) and to Buckeye at the two lowest exposure levels but not at the highest exposure level. The two adverse reproductive effects observed from avian exposure to Buckeye were an increased percentage of cracked eggs and earlier arrested embryonic development. Similarly, chronic exposure to Avio also induced earlier arrested embryonic development. These results show that the fluorine-free foams tested did cause toxicity to bobwhite quail, but whether they pose a risk at contaminated sites requires further laboratory and field study and additional exposure data. Environ Toxicol Chem 2024;43:211-221. © 2023 SETAC.

Ecotoxicity and Accumulation of Perfluorononanoic Acid in the Fathead Minnow (Pimephales promelas) and an Approach to Developing Protective Thresholds in the Aquatic Environment Through Species Sensitivity Distribution.

Per- and polyfluoroalkyl substances (PFAS) are ubiquitous in the environment. Locations where PFAS-containing aqueous film-forming foam (AFFF) has been used or accidentally released have resulted in persistently high concentrations of PFAS, including in surface water that may be adjacent to release sites. Perfluorooctane sulfonic acid (PFOS) is most frequently measured near AFFF release sites; however, other PFAS are being quantified more frequently and, of those, perfluorononanoic acid (PFNA) is common. The goal of our study was to fill data gaps on PFNA toxicity to freshwater fish using the fathead minnow (Pimephales promelas). We aimed to understand how PFNA may impact apical endpoints following a 42-day exposure to mature fish and a 21-day exposure to second-generation larval fish. Exposure concentrations were 0, 124, 250, 500, and 1000 µg/L for both adult (F0) and larval (F1) generations. The most sensitive endpoint measured was development in the F1 generation at concentrations of ≥250 µg/L. The 10% and 20% effective concentration of the tested population for the F1 biomass endpoint was 100.3 and 129.5 µg/L, respectively. These data were collated with toxicity values from the primary literature on aquatic organisms exposed to PFNA for subchronic or chronic durations. A species sensitivity distribution was developed to estimate a screening-level threshold for PFNA. The resulting hazard concentration protective of 95% of the freshwater aquatic species was 55 µg PFNA/L. Although this value is likely protective of aquatic organisms exposed to PFNA, it is prudent to consider that organisms experience multiple stressors (including many PFAS) simultaneously; an approach to understand screening-level thresholds for PFAS mixtures remains an uncertainty within the field of ecological risk assessment. Environ Toxicol Chem 2023;42:2229-2236. © 2023 SETAC.

Spatial and temporal variability of per- and polyfluoroalkyl substances (PFAS) in environmental media of a small pond: Toward an improved understanding of PFAS bioaccumulation in fish.

Per- and polyfluoroalkyl substances (PFAS) are highly fluorinated compounds with many industrial applications, for instance as ingredients in fire-suppressing aqueous film-forming foams (AFFF). Several PFAS have been demonstrated to be persistent, bioaccumulative and toxic. This study better characterizes the bioaccumulation of PFAS in freshwater fish through a spatial and temporal analysis of surface water and sediment from a stormwater pond in a former Naval air station (NAS) with historic AFFF use. We sampled environmental media from four locations twice per week for five weeks and sampled fish at the end of the sampling effort. The primary PFAS identified in surface water, sediment, and biota were perfluorooctane sulfonate (PFOS) and perfluorohexane sulfonate (PFHxS) followed by perfluorooctanoic acid (PFOA) in environmental media and perfluoroheptane sulfonate (PFHpS) in biota. We observed significant temporal variability in surface water concentrations at the pond headwaters following stochastic events such as heavy rainfall for many compounds, particularly PFHxS. Sediment concentrations varied most across sampling locations. In fish, liver tissue presented the highest concentrations for all compounds except PFHxS, which was highest in muscle tissue, suggesting the influence of fine-scale aqueous PFAS fluctuations on tissue distribution. Calculated log bioaccumulation factors (BAFs) ranged from 0.13 to 2.30 for perfluoroalkyl carboxylates (PFCA) and 0.29-4.05 for perfluoroalkane sulfonates (PFSA) and fluctuated greatly with aqueous concentrations. The variability of PFAS concentrations in environmental media necessitates more frequent sampling efforts in field-based studies to better characterize PFAS contamination in aquatic ecosystems as well as exercising caution when considering single time-point BAFs due to uncertainty of system dynamics.

Perfluoroalkyl acids in sediment and water surrounding historical fire training areas at Barksdale Air Force Base.

Perfluoroalkyl acids (PFAAs) are environmentally persistent components of surfactants that consist of fully fluorinated carbon chains and a terminal sulfonate or carboxylate polar head moiety. Due to their unique amphiphilic properties, PFAAs are used in the manufacturing of products such as aqueous film forming foams (AFFF). There is cause for concern for PFAA contamination resulting from runoff and groundwater infiltration of AFFF that were used during fire training. This study analyzed water and sediment samples that were collected over a 13-month sampling period from bayous upstream and downstream of two former fire training areas located near Barksdale Air Force Base (BAFB); the occurrence and magnitude of PFAAs supported an aquatic ecological risk assessment of potential impacts of PFAAs at the site. Liquid chromatography coupled with mass spectrometry was used for determination of 6 PFAAs listed under the third Unregulated Contaminant Monitoring Rule (UCMR 3). Total PFAA concentrations in surface water and sediment samples ranged from 0 (ND) -7.1 ng/mL and 0 (ND) -31.4 ng/g, respectively. Perfluorooctanesulfonic acid (PFOS) and perfluorooctanoic acid (PFOA) were the predominant PFAAs detected. In general, perfluorosulfonates were quantified more frequently and at higher concentrations than perfluorocarboxylates. The perfluoroalkyl chain length of PFAAs also showed significant influence on PFAA concentrations when analyzed by Spearman's rank correlation analysis. Some contamination we observed in surface water and sediment samples from reference locations could be a result of local runoff from the use of commercial products containing per- and poly-fluoroalkyl substances (PFAS), but AFFF appears to be the primary source given the close proximity of the historical fire training areas.

Comparative Toxicity of Herbicide Active Ingredients, Safener Additives, and Commercial Formulations to the Nontarget Alga Raphidocelis Subcapitata.

Chloroacetanilide herbicides are used worldwide to control weeds that affect crops such as corn, soybeans, and cotton. These herbicides are frequently paired with a "safener," which prevents herbicidal damage to the crop without diminishing weed control. Formulated herbicide products that include safeners and other ingredients are infrequently assessed for toxicity. Our goal was to understand the potential toxicity of safeners and herbicide + safener formulations relative to the toxicity of associated active ingredients. We quantified the concentration of safeners in commercially available formulations and tested effects on nontarget algae, Raphidocelis subcapitata, when exposed to individual herbicide active ingredients, safeners, and commercial formulations. The median effective concentrations (EC50s) causing 50% reduction in population growth for the herbicide active ingredients S-metolachlor and acetochlor were 0.046 and 0.003 ppm, respectively. The safeners benoxacor, AD-67, furilazole, and dichlormid were all substantially less toxic than the herbicides and were not toxic at environmentally relevant concentrations. The commercial formulations Dual II Magnum®, Me-Too-Lachlor II®, Harness®, and Surpass EC® all resulted in EC50 values that fell within the 95% confidence interval of the associated active ingredient herbicide. Interestingly, a significant increase in cell size was observed when algae were exposed to all the formulations, herbicides (acetochlor and S-metolachlor), and safener (dichlormid). The safener furilazole caused a significant decrease in cell size, whereas benoxacor and AD-67 had no observed effect on algae cell size. Significant algae cell size effects all occurred at or above the EC50 concentrations for each chemical, suggesting that other morphological effects may be occurring. Importantly, safeners in commercial formulations appeared not to impact toxicity to R. subcapitata compared with the active ingredient alone. Environ Toxicol Chem 2022;41:1466-1476. © 2022 SETAC.

Species- and Tissue-Specific Chronic Toxicity Values for Northern Bobwhite Quail (Colinus virginianus) Exposed to Perfluorohexane Sulfonic Acid and a Binary Mixture of Perfluorooctane Sulfonic Acid and Perfluorohexane Sulfonic Acid.

Per- and polyfluoroalkyl substances (PFAS) are globally distributed and present in nearly every environmental compartment. Characterizing the chronic toxicity of individual PFAS compounds and mixtures is necessary because many have been reported to cause adverse health effects. To derive toxicity reference values (TRVs) and conduct ecotoxicological risk assessments (ERAs) of PFAS-contaminated ecosystems for wildlife, species-specific PFAS chronic toxicity values (CTVs) are needed. The present study quantified PFAS residues from liver and eggs of birds chronically exposed to perfluorohexanoic acid (PFHxA) or a mixture of perfluorooctane sulfonate (PFOS) and PFHxA that produced a no-observable-adverse-effect level (NOAEL) and/or a lowest-observable-adverse-effectlevel (LOAEL). The CTVs we present are lower than those previously reported for birds and should be considered in future regulatory evaluations. From the estimated species- and tissue-specific PFAS CTVs, we found that PFOS and perfluorohexane sulfonate (PFHxS) were more bioaccumulative than PFHxA in avian tissues, but PFHxA was more toxic to reproducing birds than either PFOS or a PFOS:PFHxS mixture. We further determined that avian toxicity was not necessarily additive with respect to PFAS mixtures, which could have implications for PFAS ERAs. The PFAS LOAEL CTVs can be used to predict reproductive and possible population-level adverse health effects in wild avian receptors. Environ Toxicol Chem 2022;41:219-229. © 2021 SETAC.

Chronic Reproductive Toxicity Thresholds for Northern Bobwhite Quail (Colinus virginianus) Exposed to Perfluorohexanoic Acid (PFHxA) and a Mixture of Perfluorooctane Sulfonic Acid (PFOS) and PFHxA.

Terrestrial toxicology data are limited for comprehensive ecotoxicological risk assessment of ecosystems contaminated by per- and polyfluoroalkyl substances (PFAS) partly because of their existence as mixtures in the environment. This complicates logistical dose-response modeling and establishment of a threshold value characterizing the chronic toxicity of PFAS to ecological receptors. We examined reproduction, growth, and survival endpoints using a combination of hypothesis testing and logistical dose-response modeling of northern bobwhite quail (Colinus virginianus) exposed to perfluorohexanoic acid (PFHxA) alone and to PFHxA in a binary mixture with perfluorooctane sulfonic acid (PFOS) via the drinking water. The exposure concentration chronic toxicity value (CTV) representative of the lowest-observable-adverse effect level (LOAEL) threshold for chronic oral PFAS toxicity (based on reduced offspring weight and growth rate) was 0.10 ng/mL for PFHxA and 0.06 ng/mL for a PFOS:PFHxA (2.7:1) mixture. These estimates corresponded to an adult LOAEL average daily intake CTV of 0.0149 and 0.0082 µg × kg body weight-1 × d-1 , respectively. Neither no-observable-adverse effect level threshold and representative CTVs nor dose-response and predicted effective concentration values could be established for these 2 response variables. The findings indicate that a reaction(s) occurs among the individual PFAS components present in the mixture to alter the potential toxicity, demonstrating that mixture affects avian PFAS toxicity. Thus, chronic oral PFAS toxicity to avian receptors represented as the sum of the individual compound toxicities may not necessarily be the best method for assessing chronic mixture exposure risk at PFAS-contaminated sites. Environ Toxicol Chem 2021;40:2601-2614. © 2021 SETAC.

Toxicological Response of Chironomus dilutus in Single-Chemical and Binary Mixture Exposure Experiments with 6 Perfluoralkyl Substances.

Few studies have determined the toxicity of perfluoralkyl substances (PFAS) to aquatic invertebrates. We exposed Chironomus dilutus to 6 different PFAS to assess single-chemical toxicity and relative or proportional toxicity among substances. A 10-d range-finding test was conducted to inform 20-d assays for the following PFAS: perfluorooctane sulfonate (PFOS), perfluorooctanoic acid (PFOA), perfluorononanoic acid (PFNA), perfluorobutanesulfonic acid (PFBS), perfluorohexanesulfonic acid (PFHxS), and perfluoroheptanoic acid (PFHpA). A 20-d binary mixture study of PFOS+PFHxS followed the single-chemical tests. Measurement endpoints for 20-d tests included larval survival and biomass. Log-logistic concentration response models were used to estimate 10, 20, and 50% effect concentrations (EC20, EC50) for PFOS, PFHxS, and PFOA. Survival EC50s for PFOS, PFHxS, and PFOA were 2.49, 3860, and 192 000 µg/L, respectively, whereas survival EC20s were 1.70, 913, and 119 000 µg/L for PFOS, PFHxS, and PFOA, respectively. Biomass as a combined survival and growth endpoint resulted in EC20s of 1.89, 896, and 137 000 µg/L for PFOS, PFHxS, and PFOA, respectively. Maximum concentrations tested (no-observed-effect concentrations) for PFNA, PFBS, and PFHpA were 2 to 3 orders of magnitude greater than the PFOS EC50s and showed no toxicity to C. dilutus, even at exposure concentrations well above what would be considered environmentally relevant. The binary mixture of 2.5 µg/L PFOS+1000 µg/L PFHxS showed reduced survival compared to controls and some indication of potential additive or synergistic interaction between PFOS and PFHxS. Overall, the present study supports previous studies showing PFOS to be the most toxic PFAS to aquatic life and suggests that PFOS could be more toxic to the freshwater midge than previously reported. Environ Toxicol Chem 2021;40:2319-2333. © 2021 SETAC.

Key Considerations for Accurate Exposures in Ecotoxicological Assessments of Perfluorinated Carboxylates and Sulfonates.

Toxicity reference values for per- and polyfluoroalkyl substances (PFAS) vary even when the same test organism is studied. Although the need to confirm dosing solution concentrations is widely accepted, there are no experimental data to inform best practices when PFAS solutions are prepared. Laboratory data indicate that dissolution time of PFAS solids causes statistically significant deviations between nominal and measured concentrations. Mixing times for select PFAS varied between 2 and 5 h, depending on carbon fluorine chain-length. Environ Toxicol Chem 2021;40:677-688. © 2020 SETAC.

Assessing the Ecological Risks of Per- and Polyfluoroalkyl Substances: Current State-of-the Science and a Proposed Path Forward.

Per- and poly-fluoroalkyl substances (PFAS) encompass a large, heterogenous group of chemicals of potential concern to human health and the environment. Based on information for a few relatively well-understood PFAS such as perfluorooctane sulfonate and perfluorooctanoate, there is ample basis to suspect that at least a subset can be considered persistent, bioaccumulative, and/or toxic. However, data suitable for determining risks in either prospective or retrospective assessments are lacking for the majority of PFAS. In August 2019, the Society of Environmental Toxicology and Chemistry sponsored a workshop that focused on the state-of-the-science supporting risk assessment of PFAS. The present review summarizes discussions concerning the ecotoxicology and ecological risks of PFAS. First, we summarize currently available information relevant to problem formulation/prioritization, exposure, and hazard/effects of PFAS in the context of regulatory and ecological risk assessment activities from around the world. We then describe critical gaps and uncertainties relative to ecological risk assessments for PFAS and propose approaches to address these needs. Recommendations include the development of more comprehensive monitoring programs to support exposure assessment, an emphasis on research to support the formulation of predictive models for bioaccumulation, and the development of in silico, in vitro, and in vivo methods to efficiently assess biological effects for potentially sensitive species/endpoints. Addressing needs associated with assessing the ecological risk of PFAS will require cross-disciplinary approaches that employ both conventional and new methods in an integrated, resource-effective manner. Environ Toxicol Chem 2021;40:564-605. © 2020 The Authors. Environmental Toxicology and Chemistry published by Wiley Periodicals LLC on behalf of SETAC. This article has been contributed to by US Government employees and their work is in the public domain in the USA.

Per- and Polyfluoroalkyl Substances (PFAS) in Surface Water Near US Air Force Bases: Prioritizing Individual Chemicals and Mixtures for Toxicity Testing and Risk Assessment.

Per- and polyfluoroalkyl substances (PFAS) are a large class of persistent chemicals used for decades in industrial and commercial applications. A key challenge with regard to estimating potential risk to ecological (and human) receptors associated with PFAS exposure lies in the fact that there are many different PFAS compounds and several to many can co-occur in any given environmental sample. We applied a data science approach to characterize and prioritize PFAS and PFAS mixtures from a large dataset of PFAS measurements in surface waters associated with US Air Force Installations with a history of the use of aqueous film-forming foams (AFFFs). Several iterations of stakeholder feedback culminated in a few main points that advanced our understanding of a complex dataset and the larger ecotoxicological problem. First, perfluorooctane sulfonate (PFOS) was often a dominant PFAS in a given surface water sample, frequently followed by perfluorohexane sulfonate (PFHxS). Second, a 4-chemical mixture generally accounted for >80% of the sum of all routinely reported PFAS in a sample, and the most representative 4-chemical mixture was composed of PFOS, PFHxS, perfluorohexanoic acid (PFHxA), and perfluorooctanoic acid (PFOA). We suggest that these results demonstrate the utility of formalized data science analysis and assessment frameworks to address complex ecotoxicological problems. Specifically, our example dataset results can be used to provide perspective on toxicity testing, ecological risk assessments, and field studies of PFAS in and around AFFF-impacted sites. Environ Toxicol Chem 2021;40:871-882. © 2020 SETAC.

Species- and Tissue-Specific Avian Chronic Toxicity Values for Perfluorooctane Sulfonate (PFOS) and a Binary Mixture of PFOS and Perfluorohexane Sulfonate.

To further characterize avian toxicity to environmental levels of select per- and poly-fluoroalkyl substances (PFAS), we established species- and tissue-specific PFAS chronic toxicity values (CTVs) associated with a lowest-observable-adverse effect level (LOAEL) threshold previously established for northern bobwhite quail (Colinus virginianus) chronically orally exposed via drinking water to either perfluorooctane sulfonate (PFOS) or a simple PFAS mixture. Aided by advances in analytical techniques, the novel avian oral PFAS CTVs reported in the present study are lower than the previously reported toxicity reference values (TRVs) estimated for birds chronically exposed via feed. Thus, current avian PFOS TRVs may not be fully protective of wild avian populations at PFAS-impacted sites. Also, likely due to differences in bioavailability, bioaccessibility, and toxicokinetics among individual PFAS between oral exposure types, we found higher bioaccumulation factors in all assessed tissues from birds exposed via water versus feed. Thus, we propose that future characterization of chemical toxicity due to ingestion exposure initially include a full examination of all probable sources of oral exposure for the most accurate derivation of TRVs and a more complete picture of ecological risk. The avian PFAS LOAEL CTVs established in the present study can be modified with the use of uncertainty factors to derive site-specific avian TRVs for ecological risk assessment at PFAS-impacted sites. From differences observed in the behavior of PFOS when administered as either a single chemical or part of a binary mixture with perfluorohexane sulfonate (PFHxS), we verified that PFOS was absorbed and distributed differently when coadministered with PFHxS and that PFOS likely interacted with PFHxS differently among tissues, helping to explain the differences observed in avian toxicity between exposures. Environ Toxicol Chem 2021;40:899-909. © 2020 SETAC.

Investigating potential toxic effects of pollutants on population growth rates and probability of extinction for a representative squamate.

Chemical contamination has been suggested as an important contributing factor to reptile population declines, but direct links are rarely reported. Population modeling provides a quantitative method to understand the long-term effects of contaminants on population persistence. We created a matrix model for Sceloporus lizards and investigated hypothetical toxic effects by reducing survival and reproductive parameters by 0 to 100% in 10% increments. We report effects on population growth rate (λ) and elasticity values for each stage due to these reductions. We then incorporated stochasticity to the model to simulate the variation seen in demographic data and quantified extinction risk. The deterministic model yielded a λ of 1.07 suggesting stability in some wild Sceloporus populations. A yearly reduction of 20 to 30% in demographic parameters was needed to push λ to decline in both our deterministic and stochastic simulations. Surprisingly, our baseline stochastic simulations had a 30% extinction probability despite a stable deterministic model. We tested three adjustments to the stochastic model, (1) increased survival/fecundity parameters, (2) higher starting densities, and (3) a density-dependent juvenile survival function. The model with density-dependent juvenile growth had the lowest extinction risk. Ultimately, 20 or 30% mortality every year is likely unrealistic, but our results provide insight in linking toxicity to population effects. Ultimately, very little reduction in demographics is needed to cause declines in these populations. Our generalized models provide important tools for screening-level risk assessment of chemical contamination, especially for taxonomic groups that tend to receive less research interest.

Intraspecific interactions affect outcomes of pulse toxicity at different Daphnia magna population phases.

Traditional toxicity tests assess stressor effects on individuals, while protection goals are focused on the population-level and above. Additionally, these tests ignore common ecological factors such as resource levels and population growth phase. The objective of this research was to explore effects of - and interactions between - resource availability and stress response at the individual and population levels using Daphnia magna as a model. We hypothesized that density-dependent changes in resources at various phases of population growth would cause different population responses to the same toxicant stress. Laboratory populations of Daphnia magna were exposed to a 48-h pulse of 20 or 30 µg/l pyraclostrobin in one of four distinct phases of laboratory population cycles: growth, peak, decline, and stable. Population size and recovery were observed throughout the 51-day study. Populations exposed to pyraclostrobin during the growth phase had the least mortality and fastest recovery, while populations in the peak phase had the greatest mortality and slowest recovery. These data suggested that high density and low food at the peak phase resulted in more sensitive daphnids. To further test this hypothesis, a resource-amended acute toxicity study was conducted to quantify the effects of food resource on pyraclostrobin toxicity to Daphnia magna. Three age classes of Daphnia magna (neonate, subadult, adult) were fed low or high food levels and exposed to pyraclostrobin for 48 h. Toxicity was greater, as shown by lower 48 h LC50s, for smaller Daphnia magna age classes and lower food levels comporting results in the population study. Importantly, the acute toxicity studies generally yielded lower effect levels than the population studies suggesting that while the standard acute studies are ecologically unrealistic, they may be protective of toxicity under some circumstances. Collectively, these data point to the importance of population phase and the resource environment in modulating toxicity.

Increased temperature and lower resource quality exacerbate chloride toxicity to larval Lithobates sylvaticus (wood frog).

A chemical contaminant of growing concern to freshwater aquatic organisms, including many amphibians, is chloride ion. The salinization of freshwater ecosystems is likely caused, in part, by the application of massive amounts of road de-icing salts to roadways during winter months. The issue of freshwater salinization has become the subject of many toxicity studies and is often investigated in conjunction with other chemical stressors. However, few published studies attempt to investigate the interactions of elevated chloride concentration and increased temperature. Further, no studies have investigated the gap between the recommended feeding conditions typically used in standard toxicity tests and those that may exist in natural amphibian habitats. This study addressed the critical issues of elevated chloride, increased temperature, and variation in food quality. We conducted a 96-h acute toxicity test to investigate acute chloride toxicity as impacted by different diets, as well as a chronic toxicity test to investigate the impacts of chloride, temperature, and resource quality on the survival and development of larval Lithobates sylvaticus (wood frogs). Chloride LC50s ± 1 SE were 3769.22 ± 589.05, 2133.00 ± 185.95, and 2644.69 ± 209.73 mg Cl-/L were for non-fed, low-protein diet, and high-protein diet, respectively. For the chronic toxicity study, elevated chloride decreased tadpole survival. Increased temperature, and lower resource quality, were found negatively impacted survival of tadpoles and altered time-to-metamorphosis. This study shows that environmentally relevant concentrations of chloride, temperatures, and the protein content of the diet all exert critical effects on larval wood frogs.

Chronic Reproductive Toxicity of Perfluorooctane Sulfonic Acid and a Simple Mixture of Perfluorooctane Sulfonic Acid and Perfluorohexane Sulfonic Acid to Northern Bobwhite Quail (Colinus virginianus).

Per- and poly-fluoroalkyl substances (PFAS) are a broad class of environmentally persistent chemicals that include thousands of potentially toxic synthetic organic molecules. Some PFAS have been shown to cause adverse health effects including decreased total cholesterol, birth weight, and reproductive success in laboratory animals; however, a lack of chronic toxicity data exists for PFAS in avian ecological receptors. The present study reports on the chronic toxicity of perfluorooctane sulfonic acid (PFOS) and a mixture of PFOS and perfluorohexane sulfonic acid (PFHxS) to northern bobwhite quail (Colinus virginianus) via oral exposure from drinking water. Female weight gain was adversely affected at an average daily intake (ADI) of 3.10 × 10-3 ± 0.15 × 10-3 mg PFOS:PFHxS (1.2:1) mixture × kg-1 body weight × d-1 . Successful liberation from the shell once pipped was adversely affected at an ADI of 2.45 × 10-3 ± 0.01 × 10-3 mg PFOS x kg-1 body weight × d-1 . These values are comparatively much lower than the current dietary avian toxicity reference value (TRV) derived from birds that were exposed via feed, suggesting the need for updated avian TRVs. Relationships between test chemical (PFOS) and test substance (PFOS:PFHxS) showed that PFOS and PFHxS have possible interacting effects in avian receptors and likely differing mechanisms of toxicity depending on chemical co-occurrence and dose. Both the single-chemical and mixture exposures produced similar and possibly additive toxicity values. Environ Toxicol Chem 2020;39:1101-1111. © 2020 SETAC.

The Efficacy of the Bacillus thuringiensis israelensis Larvicide Against Culex tarsalis in Municipal Wastewater and Water from Natural Wetlands.

Water quality is a factor that is likely to interact with the activity of Bacillus thuringiensis israelensis (Bti)-based mosquito larvicides. High rates of adsorption to dense particulate matter and toxin degradation in water may be expected to reduce Bti effectiveness. Therefore, water from wastewater lagoons may serve as a protective habitat for mosquito larvae against Bti larvicides relative to natural wetlands. We conducted a study in which we exposed larvae of Culex tarsalis to various concentrations of Bti in water collected from an urban wetlands and an urban wastewater pond. We used survival analysis and a comparison of life-history characteristics to assess the effects of aquatic media on the relative efficacy of Bti and on fitness indicators. In our study, mosquitoes reared in wastewater and exposed to high (100% of minimum field application rate) and medium (50% of minimum field application rate) concentrations of Bti experienced significantly higher mortality than mosquitoes exposed to corresponding concentrations but reared in water from natural urban wetlands. In addition, females that emerged from wastewater had shorter wings than those that emerged from wetland-collected water. These effects may have been due to higher turbidity and higher ionic concentrations in wastewater than water from natural wetlands.

A Model of Culex quinquefasciatus Abundance Constructed Using Routine Surveillance and Treatment Data in Tarrant County, Texas.

Mosquito surveillance and pesticide treatment data can be combined in statistical models to provide insight into drivers of mosquito population dynamics. In cooperation with the county-based public health authority, multiple municipalities in Tarrant County, Texas, supplied surveillance and pesticide treatment data available from the 2014 mosquito season for analysis. With these data, general linear mixed modeling was used to model population dynamics of Culex quinquefasciatus, the primary vector for West Nile virus. Temporally lagged pesticide treatment information, weather data, and habitat variables were used as predictors of log + 1 transformed mosquito count data, and Akaike information criteria corrected for small sample sizes (AICc)-based model selection and multimodel averaging was used to produce a final model of mosquito abundance. The model revealed that mosquito counts were driven mainly by seasonally fluctuating temperature, precipitation, human population density, and treatment. In particular, interactions between temperature and treatment, and precipitation and human population density significantly contributed to the interpretation of the effects of the nonweather variables.