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USGS2026

Decadal shifts in groundwater age detected by environmental tracers across California, USA

Groundwater age offers important insight into recharge, storage, and contamination risk. Although models predict age changes can be driven by pumping and climate variability, direct observational evidence remains limited. Here, we analyzed paired environmental tracer suites (tritium, carbon-14, and tritiogenic helium-3) collected a decade apart from 268 wells across California to assess the prevalence of groundwater age transience. Travel-time distribution models and statistical tests indicated age transience at 29% of sites, occurring most often in agricultural regions, such as the San Joaquin Valley and Southern Coast Ranges, where large carbon-14 changes coincided with substantial nitrate and chloride shifts. Sites with tritiogenic helium-3 data showed more frequent age transience, underscoring the value of multi-tracer data sets. These results provide the first regional evidence of widespread groundwater age change and a method for detecting changing water balances with implications for groundwater sustainability and water quality.

California↗

USGS2025

System characterization report on the Environmental Mapping and Analysis Program (EnMAP)

This report addresses system characterization of the Environmental Mapping and Analysis Program hyperspectral sensor by the DLR (German Aerospace Center, ground segment project management), GFZ (Deutsches Geoforschungszentrum, science lead) and is part of a series of system characterization reports produced and delivered by the U.S. Geological Survey Earth Resources Observation and Science Cal/Val Center of Excellence. These reports present and detail the methodology and procedures for characterization; present technical and operational information about the EnMAP hyperspectral sensor; and provide a summary of test measurements, data retention practices, data analysis results, and conclusions. The Earth Resources Observation and Science Cal/Val Center of Excellence system characterization team completed data analyses to characterize the geometric (interior and exterior), and radiometric performances of the EnMAP hyperspectral sensor. Results of these analyses indicate that the Environmental Mapping and Analysis Program has a band-to-band geometric performance in the range of −0.135 to 0.15 pixel, geometric performance relative to the Operational Land Imager in the range of −27.716 meters (−0.92 pixel) to 32.892 meters (1.09 pixels) offset in comparison to Landsat 8 Operational Land Imager, offset of a radiometric comparison in the range of −0.012 to 0.020, slope of a radiometric comparison in the range of 0.947 to 1.031.

Open-File Report↗

USGS2025

Using visualization science to inform the design of environmental decision-support tools—A case study of the U.S. Geological Survey Waterwatch

Environmental decision-support tools are increasingly being used to serve both expert and non-expert audiences. Many existing tools are primarily expert-focused, and redesigning them can be challenging because experts and non-experts interact with tools differently, existing users may be resistant to changes, and there is little guidance on how to prioritize redesign efforts and demonstrate their efficacy. In this report, we present a case study of a user-centered redesign of an established environmental decision-support tool—the U.S. Geological Survey WaterWatch. WaterWatch supports flood, drought, and other water resource management decisions through the display of water levels at gages across the United States. Using a participatory process, we identified a functional change (replacing the existing rainbow colormap), created an alternative design, and tested the alternative’s usability through two general public surveys. The results showed that replacing the rainbow colormap with a more intuitive diverging colormap improves usability, regardless of the audience’s subjective preference for the rainbow color scheme. In addition, we demonstrated the importance of using legends to improve the audience’s understanding of the map symbols. This study demonstrates how user-centered design approaches can be used to inform the design of high-profile products and tools.

Scientific Investigations Report↗

USGS2025

Near-surface geophysics: Environmental applications

The field of geophysics encompasses a broad and diverse compilation of methodologies that employs principles of physics to characterize properties of earth materials within the subsurface. While geophysical methods have a long history in resource exploration and studies of Earth’s interior, the subdiscipline of “near-surface geophysics” has evolved in recent decades for examination of the shallow, near-surface environment for a range of purposes ranging from archaeological or forensic investigations to assessment of geologic, hydrologic, biologic, and geochemical properties and processes. “Environmental geophysics” are near-surface geophysical studies and methods that focus on understanding natural systems (e.g., watershed hydrology, groundwater–surface water connections, biophysical processes) as well as research pertaining to anthropogenic impacts and land management, (e.g., contamination and remediation, saltwater intrusion, agricultural practices). This field can be further subdivided into subdisciplines focused on specific topics and applications, such as water resources and hydrology (hydrogeophysics) or biologic and microbial processes (biogeophysics). Studies in environmental geophysics span a range of scales, from pore-scale laboratory tests to watershed-scale or regional field experiments. Methods vary by the nature of physics employed, the specific measurement acquired, and how that data is ultimately processed and analyzed to produce interpretable results. There exists further diversity in the acquisition logistics, geometry, and timing of data collection. Geophysical data can be collected in boreholes (one-dimensional, 1-D, vertical profiles), along survey lines (two-dimensional, 2-D, cross-sections), or in dense sensor arrays or gridded profiles (three-dimensional, 3-D, models). Regarding the temporal aspect, studies can conduct one-time geophysical surveys to obtain detailed imaging of subsurface structure or use timelapse and continuous monitoring to investigate variations in subsurface properties over time. The cumulation of all possible permutations of these factors (method, acquisition geometry, survey design, and target application) results in an immense diversity among environmental geophysical studies. Nevertheless, this field remains unified in the pursuit of understanding natural and human-impacted near-surface environments through geophysical investigations. Here we highlight some key references within environmental geophysics. Resources on geophysical theory, acquisition logistics, processing and inversion workflows, and example case studies are categorized into the most common geophysical classes within Geophysical Methods. Lastly, example references for the dominant types of applications in environmental geophysical studies are catalogued in Environmental Applications.

Book chapter↗

USGS2026

Hydrologic variability drives environmental and geospatial relationships in Smallmouth Bass (Micropterus dolomieu) distribution

Hydrologic variation is a primary driver of stream ecosystems. Changing hydrology can lead to assemblage shifts and alterations in suitable habitat for freshwater species. As climate change is predicted to alter flow patterns in addition to increasing water temperatures, insight into relationships between species occupancy, hydrology, and temperature is critical for understanding current and future distributions. We examined how hydrologic variability, temperature, and other environmental variables interact to influence Micropterus dolomieu (Smallmouth Bass) occurrence. We used Spatial Stream Network models, allowing for the incorporation of spatial autocorrelation along streams' unique dendritic network, to examine Smallmouth Bass occupancy across a range of hydrologic variation in the Ozark-Ouachita Interior Highlands, USA. Hydrologic variation was the main driver of Smallmouth Bass occurrence, with occurrence more likely in groundwater streams with low hydrologic variation and high flow permanence. For groundwater streams, occurrence was positively associated with summer stream temperature and negatively associated with annual stream temperature. As variation increased, more variables showed significant relationships with occurrence. Distance metrics were important for all models, however as hydrologic disturbance increased, flow connected distance played a lesser role and stream distance played a greater role. Hydrologic variability was the overarching determinant of Smallmouth Bass occurrence and strongly influenced the predictive importance of environmental variables and geospatial relationships. Greater hydrologic variability resulted in stronger statistical relationships between occurrence and environmental variables and an increased importance of system connectivity. As climate change alters hydrologic processes and streams become more variable, understanding and accounting for these shifting relationships is essential.

Arkansas, Kansas, Missouri, Oklahoma↗

USGS2026

Assessing environmental drivers of denitrification in restored riverine floodplains

Restoration of impaired floodplains is an increasingly prevalent strategy for alleviating water quality concerns and reducing downstream flooding at watershed scales. Floodplains temporarily store water and slow flow velocity to promote sedimentation during overbank flooding and remove inorganic nitrogen from floodwater and groundwater via denitrification. Evaluating the impacts of different restoration strategies on denitrification can inform more strategic investments into floodplain modifications that improve water quality outcomes. Our research investigates how denitrification rates in floodplains respond to environmental factors that are actionable from an engineering perspective through design and water resources management. We seasonally measured soil denitrification enzyme activity and various environmental characteristics in 4 floodplains with different restoration design and management approaches at the confluence of the Wabash and Tippecanoe Rivers in Indiana, United States. Our results showed that denitrification rates in an agricultural floodplain were significantly lower than in restored floodplains with native vegetation. Certain soil conditions characteristic of floodplain wetlands were associated with higher denitrification, particularly elevated total nitrogen, moisture, silt, and organic matter contents. Vegetation species composition was correlated with denitrification rates. This link may reflect the direct effects of vegetation on soil conditions, such as supplying labile organic carbon, or indirect effects, such as vegetation acting as an indicator of hydrologic regime and land use. Denitrification seasonally varied, peaking in winter when nitrate supply from rivers draining agricultural watersheds in the region is also high. Substrate limitation of soil denitrification enzyme activity was most significant during the summer when overbank flooding, which replenishes soil nitrogen stocks, rarely occurs. Our findings indicate that denitrification capacity will likely be maximized in riverine floodplains that are restored as wetlands with diverse native vegetation and enhanced hydrologic connectivity. Such restoration activities promote higher denitrification rates via elevated moisture, fine sediment deposition, and soil organic matter.

Tippecanoe River, Wabash River↗

USGS1999

Water-quality assessment of the New England coastal basins in Maine, Massachusetts, New Hampshire, and Rhode Island: Environmental settings and implications for water quality and aquatic biota

The New England Coastal Basins in Maine, Massachusetts, New Hampshire, and Rhode Island constitute one of 59 study units selected for water-quality assessment as part of the U.S. Geological Survey's National Water-Quality Assessment (NAWQA) program. England Coastal Basins study unit encompasses the fresh surface waters and ground waters in a 23,000 square-mile area that drains to the Atlantic Ocean. Major basins include those of the Kennebec, Androscoggin, Saco, Merrimack, Charles, Blackstone, Taunton, and Pawcatuck Rivers. Defining the environmental setting of the study unit is the first step in designing and conducting a multi-disciplinary regional water-quality assessment. The report describes the natural and human factors that affect water quality in the basins and includes descriptions of the physiography, climate, geology, soils, surface- and ground-water hydrology, land use, and the aquatic ecosystem. Although surface-water quality has greatly improved over the past 30 years as a result of improved wastewater treatment at municipal and industrial wastewater facilities, a number of water-quality problems remain. Industrial and municipal wastewater discharges, combined sewer overflows, hydrologic modifications from dams and water diversions, and runoff from urban land use are the major causes of water-quality degradation in 1998. The most frequently detected contaminants in ground water in the study area are volatile organic compounds, petroleum-related products, nitrates, and chloride and sodium. Sources of these contaminants include leaking storage tanks, accidental spills, landfills, road salting, and septic systems and lagoons. Elevated concentrations of mercury are found in fish tissue from streams and lakes throughout the study area.

Maine, Massachusetts, New Hampshire, Rhode Island↗

USGS2026

Polystyrene microplastics alter the accumulation and elimination dynamics of silver nanoparticles in Daphnia magna

Rationale Microplastics (MPs) can interact with engineered nanomaterials and alter their environmental fate and bioavailability. However, their influence on the bioaccumulation dynamics of silver nanoparticles (AgNPs) in aquatic filter feeders under environmentally relevant conditions remains poorly understood. We hypothesized that polystyrene microplastics (PS-MPs) alter the uptake, elimination and overall bioaccumulation dynamics of AgNPs in Daphnia magna . Methodology Adult D. magna were exposed to isotopically labeled citrate-coated 109 AgNPs in the presence and absence of PS-MPs. Waterborne uptake, dietary uptake, assimilation efficiency, food ingestion and elimination were quantified experimentally and incorporated into a biodynamic model to predict steady-state silver (Ag) concentrations under environmentally relevant exposure scenarios. Results PS-MPs increased waterborne Ag accumulation by approximately eightfold compared with AgNPs exposure alone. However, Ag elimination was substantially faster, with 98% of accumulated Ag eliminated after five days of depuration compared with 66% without PS-MPs. PS-MPs also reduced food ingestion rates (IRs), while assimilation efficiency remained largely unchanged. Biodynamic modeling predicted that steady-state Ag accumulation was approximately fivefold greater in the presence of PS-MPs, with waterborne exposure becoming the dominant accumulation pathway. Discussion These findings suggest that PS-MPs alter bioaccumulation dynamics of AgNPs in filter-feeding organisms by enhancing organism-associated Ag during waterborne exposure while accelerating Ag elimination, likely through particle-associated transport and gut egestion. Collectively, these results underscore the complexity of Ag bioaccumulation processes when MPs and AgNPs interact and highlight the importance of studies conducted under environmentally relevant conditions.

Environmental Chemistry↗

USGS2026

Microplastics undergo fragmentation during pressurized membrane filtration

Low-micrometer microplastics (<10 μm) have been detected in drinking water, driving growing interest in using pressure-driven membrane filtration to remove these particles and ensure drinking water safety. However, little attention is paid to the fate of microplastics concentrated in the filtration byproduct. In this study, using well-defined polystyrene (PS) and poly(methyl methacrylate) microspheres as model particles, we observed microplastic fragmentation during nanofiltration and the subsequent release of smaller fragments. After operating for 3 h at 20 bar, 67.9 ± 8.0% of the PS spheres in the concentrate, based on particles counted in selected fields of view, were transformed into fractured particles. The characteristic Raman band signals of microplastics were significantly weakened after fragmentation, leading to detection challenges. To address this, a data processing algorithm was developed to identify PS fragments as small as 1 μm. Preliminary statistical analysis reveals that within the tested pressure range, operation time has a stronger influence on fragmentation than pressure magnitude alone and that fragmentation is further governed by the intrinsic mechanical properties of the tested model polymers. This work provides direct evidence that pressure-driven membrane processes induce microplastic fragmentation and highlights the environmental risks associated with the discharge of fragmented microplastics into the concentrate.

Environmental Science & Technology Letters↗

USGS2026

Tracing mercury from land to river: Global sources, retention, and implications for sustainability

Mercury (Hg) pollution in river systems is a global sustainability challenge. Yet the transport, transformation, and retention of Hg within global rivers remain poorly quantified, particularly in regions with sparse observations such as Southeast Asia and Africa, hindering effective pollution mitigation and reinforcing geographic inequities in scientific knowledge and environmental governance. Here, we present the first global, high-resolution simulation of riverine Hg dynamics using a process-based model that traces Hg from land-based sources through river networks to the ocean. Under a realistic scenario, we estimate that ∼1900 megagrams per year (Mg/yr) of Hg enters global rivers, including 1500 Mg/yr from human-induced sources and 400 Mg/yr from soil erosion. Nearly half of this flux (∼1000 Mg/yr) is retained in reservoirs and dams, which act as major sinks. While such retention limits downstream delivery to the oceans, it also heightens in-reservoir Hg methylation risks. By bridging the gap between Hg releases and observed riverine exports, our framework offers a scalable tool for data-limited regions, promotes data access, and supports global freshwater and pollution-management strategies.

Environmental Science & Technology↗

USGS2025

Dietary bioavailability of uranium to a model freshwater invertebrate

Uranium (U) mining increases environmental exposures. Understanding how U is taken up by organisms can aid in evaluating the potential for bioaccumulation and toxicity. Although the importance of aqueous geochemical speciation is well recognized for U bioavailability after dissolved exposures, far less is known about the processes controlling U bioavailability after dietary exposures. This study characterizes the biogeochemical drivers of dietary U uptake in the freshwater snail Lymnaea stagnalis in laboratory experiments. Solids tested included benthic diatoms pre-exposed to dissolved U(VI), soils from contaminated U mine sites, and colloidal hydrous ferric oxide (HFO) synthesized in the presence of dissolved U(VI) or with U complexed by natural organic matter (NOM). Results showed that U was bioavailable from all solids. Uranium assimilation efficiency (AE), a proxy for dietary U bioavailability, varied among solids. AE was lowest for the U-contaminated soils (25 ± 17%) and highest for the U-laden diatoms (71 ± 13%). AE varied slightly among HFO preparations, suggesting modest influences of NOM and iron on U bioavailability. Increases in dietary U exposures reduced feeding rates, and the extent of feeding inhibition appeared inversely related to U bioavailability. The high U assimilation and range of bioavailability have implications for toxicity risks inferred without considering dietary uptake.

Environmental Science and Technology↗

USGS2026

Cotton farming pesticides affect ileal microbiota activity expressions of virulome but not resistome or metabolic pathways in a sedentary wild passerine

The increased use of agrochemicals to enhance crop production has had detrimental environmental effects including implication in the sharp decline of North American farmland-breeding birds. Here, using a combination of deep shotgun metatranscriptomics and pesticide exposure data, we sought to assess whether exposure to cotton ( Gossypium spp.) production had a differential effect on ileum multi-kingdom microbial activity, metabolism, anti-microbial resistance, and virulence factors of sedentary northern mockingbirds ( Mimus polyglottos ) sampled from two cotton-producing areas (16 birds in total) and one uncultivated area (7 birds) in Texas, USA. Both Shannon Index values (Adj. r 2 = 0.174, F (1,21) = 5.633, p = 0.027) and a Mantel test (Spearman ρ = 0.184, p = 0.013) supported a relationship between metabolically active microbiota Bray–Curtis dissimilarities and differences in pesticide mixtures among study areas. Virulence factor richness (Adj. r 2 = 0.182, F (1,21) = 5.890, p = 0.024), Shannon Index (Adj. r 2 = 0.231, F (1,21) = 7.612, p = 0.012), and load (sum of virulence factor abundances; Adj. r 2 = 0.160, F (1,21) = 5.194, p = 0.033) were related to total pesticide load (total quantity of pesticides). We found no pesticide effects on expression of either antimicrobial resistance genes or metabolic pathways.

Texas↗

USGS2026

Integrating habitat suitability and climate constraints to predict nonnative fish invasion risk for U.S. streams and inland lakes

Introduction: Nonnative fishes in streams and inland lakes can substantially alter aquatic ecosystems by degrading habitat conditions, competing with native species, and restructuring food webs. Identifying invasion hotspots can inform proactive management and prevention of nonnative species establishment. Methods: In this study, we identified streams and lakes across the conterminous United States that are vulnerable to invasion by 18 nonnative stream and 28 nonnative lake fish species. Habitat suitability models were developed separately for stream and lake species across nine ecoregions using natural and anthropogenic predictors, and species occurrences were predicted by integrating model results with climate match scores based on comparisons between environmental conditions in species’ native and potential nonnative ranges. We classified areas with both high habitat suitability and strong climate similarity as having high invasion potential and generated hotspot maps by stacking species-level predictions. Results: Florida exhibits the highest invasion risk for both habitat types. Stream hotspots were concentrated near major metropolitan areas, whereas lake hotspots were more widely distributed across the Great Lakes basin and the northeastern U.S. Twelve species were predicted to invade both habitat types, with goldfish ( Carassius auratus ) and pirapitinga ( Piaractus brachypomus ) showing particularly high invasion potential. Discussion: These findings provide a comprehensive assessment of nonnative freshwater fish invasion risk and support targeted monitoring and management strategies.

conterminous United States↗

USGS2025

Evaluation of daily stream temperature predictions (1979-2021) across the contiguous United States using a spatiotemporal aware machine learning algorithm

Stream temperature controls a variety of physical and biological processes that affect ecosystems, human health, and economic activities. We used 42 years (1979–2021) of data to predict daily summary statistics of stream temperature across >50,000 stream reaches in the contiguous United States using a recurrent graph convolution network. We comprehensively documented the performance – both across all reaches and by stream type (e.g., reservoir or groundwater influence) – as a baseline for future improvement. The model showed reach-level RMSE of <2 °C with 90 % prediction intervals that contain 90.7 % of observations. We also assessed how the model captured variability in ecologically relevant metrics (e.g., R 2 for annual 7-day maximum = 0.76; R 2 for days exceeding 25 °C = 0.75). This model does not outperform state-of-the-art machine learning efforts (e.g., RMSE ≤1.5 °C) due to a limited input set but does provide the most spatially complete modeling to date to support water availability assessments.

contiguous United States↗

USGS2026

Multiple machine-learning estimation of groundwater levels and trends for the regional Mississippi River Valley alluvial aquifer

The Mississippi River Valley alluvial aquifer provides irrigation, public, and domestic water supplies across the south-central United States. Declining groundwater levels require improved characterization of changing conditions. Traditional potentiometric-surface mapping does not use all available water-level data or quantify uncertainty. To address these limitations, we developed a data-driven multiple machine-learning (MML) framework delivered through two open-source R packages. The covMRVAgen1 software assembles covariates to 155,960 monthly groundwater levels from 57,695 wells; the mmlMRVAgen1 software trains Cubist and Random Forest models, blends them, and makes 1-kilometer gridded predictions of monthly potentiometric surfaces for the period January 1980–December 2022. The MML approach provides a methodological foundation for region-scale spatiotemporal groundwater prediction and uncertainty quantification, generating 90-percent prediction limits with appropriate empirical coverage. Model performance is acceptable, with a root-mean-square error of about 4.2 feet, standard deviation of 24.82 feet, and a normalized Nash–Sutcliffe efficiency of 0.973.

Arkansas, Illinois, Louisiana, Mississippi, Missou↗

USGS2025

Climate-driven sulfate export in alpine watersheds may stimulate methylmercury production

Climate change is increasing sulfate export and changing wetland extent in mountain regions. These changes may increase microbially mediated production of the neurotoxic substance methylmercury due to enhanced sulfate metabolism in mountain environments. Here, we assess methylmercury concentrations and formation rates across high-elevation wetlands in the Colorado Rocky Mountains. We also investigate sulfate controls on methylmercury production within subalpine peatlands by amending soils with sulfate to mimic increased stream export of sulfate from the alpine zone and measuring methylmercury formation rates for different sulfate treatments. We found that subalpine peatlands have statistically significant higher methylmercury concentrations and formation rates compared to alpine, mineral-soil wetlands. Methylmercury production in subalpine peatlands also increased significantly ( p < 0.05) following sulfate additions; the highest rates occurred in sediments with intermediate extractable sulfate concentrations (∼0.60–1.4 mg sulfate g −1 dry soil). Our study is the first to identify soil sulfate-related thresholds for methylmercury production and sulfate-limitation of methylmercury production in subalpine peatlands. These findings highlight important linkages between climate-driven mineral weathering and mercury cycling in mountain regions globally.

Colorado↗

USGS1999

Environmental setting and water-quality issues in the lower Tennessee River basin

The goals of the National Water-Quality Assessment Program are to describe current water-quality conditions for a large part of the Nation's water resources, identify water-quality changes over time, and identify the primary natural and human factors that affect water quality. The lower Tennessee River Basin is one of 59 river basins selected for study. The water-quality assessment of the lower Tennessee River Basin study unit began in 1997. The lower Tennessee River Basin study unit encompasses an area of about 19,500 square miles and extends from Chattanooga, Tennessee, to Paducah, Kentucky. The study unit had a population of about 1.5 million people in 1995. The study unit was subdivided into subunits with relatively homogeneous geology and physiography. Subdivision of the study unit creates a framework to assess the effects of natural and cultural settings on water quality. Nine subunits were delineated in the study unit; their boundaries generally coincide with level III and level IV ecoregion boundaries. The nine subunits are the Coastal Plain, Transition, Western Highland Rim, Outer Nashville Basin, Inner Nashville Basin, Eastern Highland Rim, Plateau Escarpment and Valleys, Cumberland Plateau, and Valley and Ridge.The lower Tennessee River Basin consists of predominantly forest (51 percent) and agricultural land (40 percent). Activities related to agricultural land use, therefore, are the primary cultural factors likely to have a widespread effect on surface- and ground-water quality in the study unit. Inputs of total nitrogen and phosphorus from agricultural activities in 1992 were about 161,000 and 37,900 tons, respectively. About 3.7 million pounds (active ingredient) of pesticides was applied to crops in the lower Tennessee River Basin in 1992. State water-quality agencies identified nutrient enrichment and pathogens as water-quality issues affecting both surface and ground water in the lower Tennessee River Basin. Water-quality data collected by State and Federal agencies between 1980 and 1996 were summarized to characterize surface- and ground-water quality of the subunits with respect to these issues. Median concentrations of nitrogen species generally were less than 1 milligram per liter in surface and ground water in all subunits, and were highest throughout the subunits that had the largest percentages of agricultural land use. Median phosphorus concentrations also were less than 1 milligram per liter in all subunits. Phosphatic limestones present in two subunits had a larger effect on phosphorus concentrations in surface and ground water than did the amount of agricultural land use in these subunits. Median counts of fecal coliform were higher in surface water than in ground water in all subunits. The highest median counts in surface water were in the Valley and Ridge (7,500 colonies per 100 milliliters) and the Outer Nashville Basin subunits (5,000 colonies per 100 milliliters). Highest median counts in ground water were in the Inner and Outer Nashville Basin subunit. Natural setting likely has an important effect with respect to fecal contamination of surface and ground water in the lower Tennessee River Basin.

Alabama, Georgia, Kentucky, Mississippi, Tennessee↗

USGS2026

PFAS mixture composition and internal exposure profiles shape biological responses under field-realistic exposure

Per- and polyfluoroalkyl substances (PFAS) occur as complex mixtures, yet mixture-dependent biological effects under environmentally realistic exposure conditions remain poorly understood. We conducted multiyear (2018, 2019, 2021) continuous-flow, field-based exposures of male fathead minnows ( Pimephales promelas ) using a low-PFAS reference well (REF; sum of measured PFAS (∑PFAS) 0.1–0.2 μg L –1 ) and PFAS-contaminated groundwater from a fire-training area (FTA) at Joint Base Cape Cod, Massachusetts. Dilution treatments enabled separation of concentration and mixture effects. Groundwater from well FTA1 was perfluorooctanesulfonate (PFOS) dominated (∑PFAS 10–31 μg L –1 ), whereas groundwater from well FTA2 had higher concentrations and was enriched in perfluorooctanoate (PFOA) and diverse precursors (∑PFAS ∼ 80 μg L –1 ). Despite comparable plasma ∑PFAS and PFOS in FTA1–100% and FTA2–50% on day-7, cumulative mortality reached approximately 35% in FTA2–50% and 17% in FTA2–100%, and secondary sex trait expression was reduced by approximately 65–80% relative to REF, whereas sperm motility effects were mixture- and time-dependent. Plasma PFAS profiles were dominated by perfluorohexanesulfonate (PFHxS), PFOS, and sulfonamide precursors. Liver transcriptomics from REF-acclimated fish revealed robust disruption of metabolic, mitochondrial, and endocrine pathways that link PFAS mixture chemistry and internal exposure profiles to organismal outcomes, and testis transcriptomics provided complementary insight into reproductive impairment. These results indicate that PFAS mixture composition and internal exposure profiles, including precursor-associated differences, are important determinants of ecotoxicological outcomes under field-realistic conditions.

Massachusetts↗