Discussion. Has the human population become a sentinel for the adverse effects of PFAS contamination on wildlife health and endangered species?

Global contamination with per- and polyfluoroalkyl substances (PFAS) poses a threat to both human health and the environment, with significant implications for ecological conservation policies. A growing list of peer-reviewed publications indicates that PFAS can harm wildlife health and that the adverse effects associated with PFAS exposure in wildlife are in concordance with human epidemiological studies. The correlation of cross-species data supports a unique perspective that humans can be regarded as a sentinel for PFAS effects in other species. The health harms due to PFAS are potentially most concerning for populations of endangered and threatened species that are simultaneously exposed to PFAS and other toxic pollutants, and also face threats to their survival due to habitat loss, degradation of ecosystems, and over-harvesting. Human epidemiological studies on the PFAS doses associated with health harm present a rich source of information about potential impacts on wildlife health due to PFAS. Our analysis suggests that national and international efforts to restrict the discharges of PFAS into the environment and to clean up PFAS-contaminated sites present an opportunity to protect wildlife from chemical pollution and to advance species conservation worldwide.

Locally caught freshwater fish across the United States are likely a significant source of exposure to PFOS and other perfluorinated compounds.

Per- and polyfluoroalkyl substances, or PFAS, gained significant public and regulatory attention due to widespread contamination and health harms associated with exposure. Ingestion of PFAS from contaminated food and water results in the accumulation of PFAS in the body and is considered a key route of human exposure. Here we calculate the potential contribution of PFOS from consumption of locally caught freshwater fish to serum levels. We analyzed data for over 500 composite samples of fish fillets collected across the United States from 2013 to 2015 under the U.S. EPA's monitoring programs, the National Rivers and Streams Assessment and the Great Lakes Human Health Fish Fillet Tissue Study. The two datasets indicate that an individual's consumption of freshwater fish is potentially a significant source of exposure to perfluorinated compounds. The median level of total targeted PFAS in fish fillets from rivers and streams across the United States was 9,500 ng/kg, with a median level of 11,800 ng/kg in the Great Lakes. PFOS was the largest contributor to total PFAS levels, averaging 74% of the total. The median levels of total detected PFAS in freshwater fish across the United States were 278 times higher than levels in commercially relevant fish tested by the U.S. Food and Drug Administration in 2019-2022. Exposure assessment suggests that a single serving of freshwater fish per year with the median level of PFAS as detected by the U.S. EPA monitoring programs translates into a significant increase of PFOS levels in blood serum. The exposure to chemical pollutants in freshwater fish across the United States is a case of environmental injustice that especially affects communities that depend on fishing for sustenance and for traditional cultural practices. Identifying and reducing sources of PFAS exposure is an urgent public health priority.

Investigating Molecular Mechanisms of Immunotoxicity and the Utility of ToxCast for Immunotoxicity Screening of Chemicals Added to Food.

The development of high-throughput screening methodologies may decrease the need for laboratory animals for toxicity testing. Here, we investigate the potential of assessing immunotoxicity with high-throughput screening data from the U.S. Environmental Protection Agency ToxCast program. As case studies, we analyzed the most common chemicals added to food as well as per- and polyfluoroalkyl substances (PFAS) shown to migrate to food from packaging materials or processing equipment. The antioxidant preservative tert-butylhydroquinone (TBHQ) showed activity both in ToxCast assays and in classical immunological assays, suggesting that it may affect the immune response in people. From the PFAS group, we identified eight substances that can migrate from food contact materials and have ToxCast data. In epidemiological and toxicological studies, PFAS suppress the immune system and decrease the response to vaccination. However, most PFAS show weak or no activity in immune-related ToxCast assays. This lack of concordance between toxicological and high-throughput data for common PFAS indicates the current limitations of in vitro screening for analyzing immunotoxicity. High-throughput in vitro assays show promise for providing mechanistic data relevant for immune risk assessment. In contrast, the lack of immune-specific activity in the existing high-throughput assays cannot validate the safety of a chemical for the immune system.

Do flame retardant concentrations change in dust after older upholstered furniture is replaced?

Upholstered furniture has been a major source of chemical flame retardant (FR) exposures in US homes since the 1970s. FRs are a large group of chemicals, many of which are associated with adverse health effects, including cancer, reproductive toxicity, and neurotoxicity. California homes have some of the highest dust concentrations of FRs, due to Technical Bulletin 117 (TB117), California's outdated flammability standard for furniture foam that was generally followed across the US and Canada. In 2014, this standard was updated to a smolder standard for furniture fabric called TB117-2013, and it is no longer reliant on FRs. This update provided an opportunity to measure differences in FR dust levels in California homes before and after residents replaced older upholstered furniture, or its foam, with products that met the new standard and were expected to be FR-free. We collected dust from homes of participants who had plans to replace older upholstered furniture, or furniture foam, with FR-free options. We returned for follow-up dust collection six, 12, and 18 months following replacement. Concentrations of three polybrominated diphenyl ethers (PBDEs) (BDE-47, BDE-99, BDE-100), three chlorinated organophosphate ester FRs (tris(2-chloroethyl) phosphate (TCEP), tris(2-chloroisopropyl) phosphate (TCIPP), and tris(1,3-dichloroisopropyl) phosphate (TDCIPP)), and one aryl organophosphate ester FR triphenyl phosphate (TPHP), were widely detected in participant homes. All measured FRs decreased in nearly all homes after the older upholstered furniture was replaced. The decreases in FRs were significant in both homes that replaced entire pieces of furniture and those that replaced only the furniture foam. This study demonstrates that replacing older upholstered furniture or foam significantly reduces concentrations of a range of FRs in the home. Foam replacement offers a potentially more economic alternative that produces a lower volume of waste.

Asbestos Contamination in Talc-Based Cosmetics: An Invisible Cancer Risk.

The lack of regulation and adequate testing of talc-containing personal care products in the U.S. has resulted in the contamination of cosmetics with asbestos. As such, the true exposure of consumers to asbestos is poorly characterized and likely underestimated. In this study, transmission electron microscopy analysis revealed that 3 of 21 powder-based cosmetic products tested were contaminated with amphibole asbestos. One of these contaminated products is expressly marketed for use by children. The presence of asbestos found in products demonstrates the urgency to revise cosmetics policy. Further, talc-based cosmetics may be an overlooked and difficult to characterize source of exposure to asbestos, a known carcinogen.

Disposal of products and materials containing per- and polyfluoroalkyl substances (PFAS): A cyclical problem.

Per- and polyfluoroalkyl substances (PFAS), highly stable and persistent chemicals used in numerous industrial applications and consumer goods, pose an exceptionally difficult challenge for disposal. Three approaches are currently available for PFAS wastes: landfilling, wastewater treatment and incineration. Each disposal approach can return either the original PFAS or their degradation products back to the environment, illustrating that the PFAS problem is cyclical. Landfilling and wastewater treatment do not destroy PFAS and simply move PFAS loads between sites. Consumer products and various materials discarded in landfills leach PFAS over time, and landfill leachate is commonly sent to wastewater treatment plants. From wastewater treatment plants, PFAS are carried over to sludge and effluent. Sewage sludge can be landfilled, incinerated, or applied on agricultural fields, and PFAS from treated sludge (biosolids) can contaminate soil, water, and crops. Incineration of PFAS-containing wastes can emit harmful air pollutants, such as fluorinated greenhouse gases and products of incomplete combustion, and some PFAS may remain in the incinerator ash. Volatile PFAS are emitted into the air from landfills and wastewater treatment plants, and research is urgently needed on the potential presence of PFAS compounds in air emissions from commercially run incinerators. Monitoring of waste streams for PFAS, stopping PFAS discharges into water, soil and air and protecting the health of fence-line communities close to the waste disposal sites are essential to mitigate the impacts of PFAS pollution on human health.

Applying a cumulative risk framework to drinking water assessment: a commentary.

The health risks of drinking water contaminants and the economic benefits of drinking water standards are typically assessed one chemical at a time, an approach that misses the health impacts of co-occurring contaminants in drinking water. In contrast, a cumulative risk framework has become common in air quality evaluations such as the U.S. Environmental Protection Agency's National Air Toxics Assessment. We posit that the drinking water field would benefit from making the transition to a unified assessment framework for multiple contaminants that can overcome the long-standing challenge of treating cancer and non-cancer contaminants separately. Here we present a cumulative risk methodology that combines a risk-based cancer metric with a weighted health indicator index for non-cancer contaminants and incorporates disability weights from the Global Burden of Disease study. Our methodology generates a numeric toxicity score reflecting the potential health impacts for the sum of contaminants present in each sample of drinking water. Further research is needed to refine the risk and toxicity parameters for specific contaminants and to address the mode of interaction between co-occurring chemicals. As this cumulative risk model goes through future refinements, we anticipate that it would provide information that can help communities and policy makers evaluate different options for drinking water treatment.

Influence of hardness on the bioavailability of silver to a freshwater snail after waterborne exposure to silver nitrate and silver nanoparticles.

The release of Ag nanoparticles (AgNPs) into the aquatic environment is likely, but the influence of water chemistry on their impacts and fate remains unclear. Here, we characterize the bioavailability of Ag from AgNO(3) and from AgNPs capped with polyvinylpyrrolidone (PVP AgNP) and thiolated polyethylene glycol (PEG AgNP) in the freshwater snail, Lymnaea stagnalis, after short waterborne exposures. Results showed that water hardness, AgNP capping agents, and metal speciation affected the uptake rate of Ag from AgNPs. Comparison of the results from organisms of similar weight showed that water hardness affected the uptake of Ag from AgNPs, but not that from AgNO(3). Transformation (dissolution and aggregation) of the AgNPs was also influenced by water hardness and the capping agent. Bioavailability of Ag from AgNPs was, in turn, correlated to these physical changes. Water hardness increased the aggregation of AgNPs, especially for PEG AgNPs, reducing the bioavailability of Ag from PEG AgNPs to a greater degree than from PVP AgNPs. Higher dissolved Ag concentrations were measured for the PVP AgNPs (15%) compared to PEG AgNPs (3%) in moderately hard water, enhancing Ag bioavailability of the former. Multiple drivers of bioavailability yielded differences in Ag influx between very hard and deionized water where the uptake rate constants (k(uw), l g(-1) d(-1) ± SE) varied from 3.1 ± 0.7 to 0.2 ± 0.01 for PEG AgNPs and from 2.3 ± 0.02 to 1.3 ± 0.01 for PVP AgNPs. Modeling bioavailability of Ag from NPs revealed that Ag influx into L. stagnalis comprised uptake from the NPs themselves and from newly dissolved Ag.

Does water chemistry affect the dietary uptake and toxicity of silver nanoparticles by the freshwater snail Lymnaea stagnalis?

Silver nanoparticles (AgNPs) are widely used in many applications and likely released into the aquatic environment. There is increasing evidence that Ag is efficiently delivered to aquatic organisms from AgNPs after aqueous and dietary exposures. Accumulation of AgNPs through the diet can damage digestion and adversely affect growth. It is well recognized that aspects of water quality, such as hardness, affect the bioavailability and toxicity of waterborne Ag. However, the influence of water chemistry on the bioavailability and toxicity of dietborne AgNPs to aquatic invertebrates is largely unknown. Here we characterize for the first time the effects of water hardness and humic acids on the bioaccumulation and toxicity of AgNPs coated with polyvinyl pyrrolidone (PVP) to the freshwater snail Lymnaea stagnalis after dietary exposures. Our results indicate that bioaccumulation and toxicity of Ag from PVP-AgNPs ingested with food are not affected by water hardness and by humic acids, although both could affect interactions with the biological membrane and trigger nanoparticle transformations. Snails efficiently assimilated Ag from the PVP-AgNPs mixed with diatoms (Ag assimilation efficiencies ranged from 82 to 93%). Rate constants of Ag uptake from food were similar across the entire range of water hardness and humic acid concentrations. These results suggest that correcting regulations for water quality could be irrelevant and ineffective where dietary exposure is important.

Induction of reactive oxygen species in chlamydomonas reinhardtii in response to contrasting trace metal exposures.

The toxicity of metals to organisms is, in-part, related to the formation of reactive oxygen species (ROS) in cells and subsequent oxidative stress. ROS are by-products of normal respiration and photosynthesis processes in organisms, but environmental factors, like metal exposure, can stimulate excess production. Metals involved in several different mechanisms such as Haber-Weiss cycling and Fenton-type reactions can produce ROS. Some metals, such as Cd, may contribute to oxidative stress indirectly by depleting cellular antioxidants. We investigated the measurement of ROS as a sensitive biomarker of metal toxicity (that could possibly be implemented in a biotic ligand model for algae) and we compared ROS induction in response to several contrasting transition metals (Cu, V, Ni, Zn, and Cd). We also compared the ROS response to glutathione and growth toxicity endpoints measured in a previous study. The cell-permeable dye, 2'7'dichlorodihydrofluorescein diacetate, was used as a probe to detect formation of ROS in Chlamydomonas reinhardtii cells. Metal-exposed cells were incubated with the fluorescent dye in a 96-well plate and monitored over 5.5 h. A dose-response of ROS formation was observed with Cu exposure in the range of 20-500 nM. Cu produced more ROS compared with either Zn or Cd (both nonredox active metals). The redox-active metal V produced increased ROS with increased concentration. The measurement of ROS may be a useful indicator of Cu toxicity, but the signal to noise ratio was better for the glutathione endpoint assay.

Relationships between surface-bound and internalized copper and cadmium and toxicity in Chlamydomonas reinhardtii.

In the present study, the adsorption and uptake of copper (Cu) and cadmium (Cd) in Chlamydomonas reinhardtii were examined to establish fundamental toxicity relationships to glutathione and cell-growth endpoints. Establishing these fundamental relationships of metal accumulation and toxicity metrics is necessary to subsequently implement an algal biotic ligand model. The glutathione response was similar to the response measured from growth endpoints for both internal and adsorbed Cu, indicating that glutathione may be a useful biomarker of toxicity. The glutathione response with Cd contrasted markedly with that observed with Cu and was therefore observed to be a metal-specific biomarker. The density of sites binding metals and the related stability constants for the algal cell surface were also determined. Short exposures to metals (2 h) were conducted, and we determined 6.0 × 10(-6) mol/g sites binding Cu and 2.0 × 10(-6) mol/g sites binding Cd and conditional stability constants as log K' = 7.2 and log K' = 6.7 for Cu and Cd, respectively. Experiments were also conducted to determine the effect on toxicity endpoints of varying nitrate concentrations and different humic acids (HA) in the exposure media. Varying nitrate concentrations did not have an effect on cell growth over 24 h. The surface-adsorbed Cu measurements from the experiments with HA depended on the type and concentration of HA.

Differential effects of copper and cadmium exposure on toxicity endpoints and gene expression in Chlamydomonas reinhardtii.

The toxicity of cadmium to aquatic organisms is well known, but the mechanisms of toxicity are not as clearly understood. In the present study, Cd bioassay experiments incorporating both traditional endpoints and novel thiol-based endpoints were conducted with Chlamydomonas reinhardtii. The results were compared with results from previous bioassay experiments to probe the apparent contrasting biochemical mechanisms of toxicity of copper and cadmium as expressed in cellular glutathione and the glutathione cycle. Total glutathione and reduced to oxidized glutathione ratio (GSH/GSSG) measurements were remarkably different in Cd- compared with Cu-exposed cells. Whereas total glutathione in cells decreased with increasing Cu concentration, Cd caused dramatic increases. Total glutathione increased by 4.5-fold with 80 nM Cd treatment over concentrations in Cd-free controls. Glutathione reductase (GR) enzyme activity was positively correlated (r(2) (Cu) = 0.96, r(2) (Cd) = 0.85) with glutathione concentrations for both metals. Measurements of mRNA for GR were increased 2-fold in response to Cd exposure (80 nM) and correlated well with GR enzyme activity. Glutathione concentrations and GR enzyme activity are useful endpoints for both Cu and Cd toxicity in algae, even though the metals elicit opposing responses. We conclude that Cu decreases glutathione concentrations by inhibiting GR enzyme activity. In contrast, Cd stimulates GR enzyme activity and increases glutathione concentrations as cells respond to Cd-induced stress by producing increased antioxidant capacity. The present study demonstrates that determining the glutathione response in cells is important for understanding the metal-specific mechanisms of toxicity and that these associated novel endpoints may be useful metrics for accurately predicting toxicity.

Analysis of glutathione endpoints for measuring copper stress in Chlamydomonas reinhardtii.

Glutathione (GSH) is the most abundant nonprotein thiol in eukaryotic cells and it protects cells by functioning as an antioxidant and a metal-binding ligand. Because glutathione readily undergoes oxidation-reduction reactions to combat oxidative stress, intracellular ratios of the reduced (GSH) to the oxidized (GSSG) forms of glutathione may serve as an important biomarker of exposure and effect of trace metals in eukaryotic cells. We compared sensitivity of glutathione ratios in the freshwater alga Chlamydomonas reinhardtii to the traditional endpoints of cell growth rates and chlorophyll a following exposure to Cu for periods of 6 and 24 h. A response of the GSH:GSSG ratio to Cu concentration was observed at Cu levels of 40 and 80 nM after exposure for both 6 and 24 h. The concentration of total GSH at 24 h was roughly half the value at 6 h after exposure to either 40 or 80 nM Cu. A response for cell growth rate was observed only at 24 h, whereby the average specific growth rate decreased from about 1.1 to 0.4 d(-1). The total Cu concentrations eliciting a cell response of 50%, effect concentrations (EC50s), after 24 h of exposure were similar (49.2, 49.8, and 38.2 nM Cu) and not significantly different for GSH:GSSG ratio, GSH levels, and specific growth, respectively. Total cell-associated Cu concentrations after exposure for 24 h were calculated from the EC50 endpoints and ranged from 13.3 to 17.0 fg/cell. Overall, thiol ratios were indicative of toxicity resulting from exposure to Cu, but precision may be greater for the cell growth rate endpoints.