Science of The Total Environment

Mercury biomagnification in Amazonian rivers is usually assessed through fishes, yet contaminants can also move across riparian food webs through multiple energy pathways. We evaluated how total mercury (THg) is structured across the upper Caqueta River basin, a mining-impacted river floodplain system in the Colombian Amazon. Using stable carbon and nitrogen isotopes, trophic guild classification, biomagnification models, and Bayesian mixing models, we integrated basal resources, macroinvertebrates, fishes, and birds to link mercury exposure with food-web architecture. THg increased with trophic enrichment in fishes, birds, and the combined food web, indicating biomagnification across aquatic and aquatic-associated consumers. Reconstructed pathways showed that phytoplankton and detritus support intermediate fish consumers, which in turn connect basal production to carnivore piscivorous fishes and aquatic predatory birds. Avian results suggest that mercury exposure is not restricted to aquatic predators, as hummingbirds showed unexpectedly elevated THg values that may reflect additional riparian or arthropod-mediated pathways. Overall, mercury transfer in the upper Caqueta emerges from the interaction between trophic position and energy-flow structure, highlighting the need to evaluate contamination magnification through integrated aquatic riparian food webs rather than single consumer groups.

Background: Guanica, Puerto Rico, has one of the highest concentrations of environmental polychlorinated biphenyls (PCBs). This PCB contamination coupled with persistent poverty, population loss, and repeated environmental disasters has taken a heavier toll on Guanica community. Our environmental sampling has documented high levels of PCBs in sediment, air and fish, but far less is known about how residents themselves understand exposure, interpret health risk, and weigh contamination against the other pressures of daily life. Methods: We used a qualitative descriptive design nested within the larger HELP Study. This paper presents the final analysis of all qualitative data collected in Guanica: ten sessions comprising five focus groups and five interviews with fishermen (N = 31), conducted between November 2024 and October 2025 in familiar community settings. Sessions followed a semi-structured guide translated from English to Spanish and back-translated for accuracy. Recordings were transcribed in Spanish, verified against the audio, and analyzed in NVivo using codebook-style thematic analysis; three bilingual researchers coded collaboratively and resolved disagreements by consensus. Results: Five themes emerged from our analysis concerning how residents and fishermen understand and navigate PCB contamination: (1) perceived community decline shapes how residents prioritize environmental risks; (2) limited knowledge, misinformation, and communication gaps affect understanding of PCB contamination; (3) cancer concerns are high, but links to PCB exposure remain unclear; (4) PCB contamination is seen as a threat to livelihoods, especially fishing and tourism; and (5) deep mistrust and a desire for accountability influence perceptions of environmental interventions. Participants described contamination not as an isolated hazard but as one strand of a longer history of economic decline, disaster, and institutional neglect. Conclusions: Social, economic, and environmental conditions together shape how Guanica residents perceive contamination of their environment. Meaningful progress requires sustained community involvement, support for local initiatives, development of a Community Health Worker program, and expanded partnerships with local institutions and schools to close communication gaps and promote long-term resilience to address the health effects of PCB contamination in Guanica.

This study assesses the spatial dynamics of heavy metals and associated trace elements in the water and sediment of the Slana River approximately one and half year after an environmental accident and river contamination caused by mining effluent. Using PCA and standardized environmental indices (CF, PLI), we modelled the distribution of contaminants across the macro-gradient of the river and within the micro-gradient surrounding the source of pollution. We found that although the situation in the water column has improved and stabilized, the sediment remains the primary reservoir of trace elements, including toxic arsenic, with the highest concentrations recorded near the source and in its immediate vicinity. Although the concentration of primary mining indicators decreases relatively rapidly along the stream, this is systematically offset by the accumulation of other elements, including sulphur and aluminium. As a result of these geochemical processes, the total pollution load index (PLI) remains statistically unchanged along the entire stream, but indicates a chronic, spatially uniform (PLI = 1.06-1.16) toxicological burden on the ecosystem.

Long-term PM2.5 exposure is associated with increased mortality and is usually represented by annual mean concentration, which does not capture how concentrations are distributed across days. We tested whether the prespecified PM2.5 magnitude-rank index (PMRI), a summary of within-year daily concentration structure, improves mortality prediction beyond the annual mean. Daily modeled PM2.5 estimates were linked to age-adjusted mortality rates for 27,289 county-years from 3,064 U.S. counties during 2003-2011. PMRI is the largest k for which at least k days reached k micrograms per cubic meter. County-grouped 10-fold cross-validation compared models with annual mean alone versus annual mean plus PMRI, with additional geographic, temporal, cause-specific, and specification analyses. Annual mean PM2.5 had the highest standalone predictive performance. Adding PMRI increased county-held-out R-squared from 0.0802 to 0.1391 (increase in R-squared = 0.0589; 95% CI, 0.0499-0.0677) and reduced RMSE and MAE. Improvement persisted across geographic and temporal validations and all six cause-specific mortality outcomes. Within-year PM2.5 exposure structure therefore contains reproducible mortality-predictive information not captured by annual mean concentration.

Hormesis is an important consideration in chemical risk assessment across most aquatic environments. Evidence suggests that hormetic responses are shaped by organismal physiological processes. Accordingly, predictions of hormesis under environmentally realistic conditions should account for physiological traits. Yet the role of these traits remains largely unexplored. This study investigated how baseline growth rate modulates antibiotic responses in the harmful cyanobacterium Microcystis and the underlying mechanisms. Results showed that at an antibiotic concentration of 200 ng L-1 (representing environmental concentration levels), slow-growing Microcystis populations exhibited an obvious growth enhancement more than 4-fold greater than that of fast-growing populations, and this enhancement was positively correlated with cellular reserves. A baseline growth rate below 0.19 day-1 emerged as the threshold below which hormetic responses were consistently observed. This study provides pioneering insights into how organisms' physiological states influence pollutant-induced hormetic responses in aquatic environments. Our findings point to a central principle for extrapolation that laboratory-derived toxicological responses may not translate directly to the field unless key physiological traits of the organisms are explicitly considered. In this regard, baseline growth rate and energy reserves may be key physiological traits for reliably predicting toxic effects at the population level. This study also makes an initial attempt to conceptualize a physiology-anchored framework for strengthening risk assessment of low-level emerging contaminants.

Subsurface tile drainage rapidly transports nutrients and biological contaminants to surface waters, yet multi-year evaluations of drainage water management (DWM) effects on nutrients, Escherichia coli, and antibiotic resistance genes (ARGs) remain limited. A paired-field design with high-frequency soil moisture monitoring was used to quantify water yield, dissolved nitrogen, phosphorus, and Escherichia coli (E. coli) over seven and eight water years, and ARGs (tetM, tetW, ermB, sul1) over two years in southern Michigan. Soil moisture at tile depth (~75 cm) was higher under controlled drainage, confirming that DWM increases field water-holding capacity. Compared to the free drainage field, DWM reduced cumulative tile discharge by 43% and annual nutrient yields by 46-83% for nitrate and 12-82% for soluble reactive phosphorus, but effects on ammonium were variable. Average daily nutrient yields declined by ~50% over the study period, driven primarily by managed periods. Nutrient concentrations did not differ between treatments, indicating that yield reductions resulted from decreased water loss. Mean daily E. coli yields declined by 71% over the study period, and concentrations were 46% lower under controlled drainage. Controlled drainage reduced tetM and sul1 ARG concentrations and yields during managed periods and throughout the study; however, sensitivity analyses showed that these effects were strongly influenced by a high-precipitation event. After excluding the event, differences in ARG concentrations were no longer detected, although ARG yields remained 43% lower during managed periods. These findings highlight the potential for DWM to mitigate nutrient, microbial, and antibiotic resistance transport from agricultural drainage systems.

Urban green infrastructure is increasingly promoted as a public-health intervention, yet the biological features that determine exposure to potentially beneficial plant-derived volatile compounds remain poorly understood. Here we quantified airborne terpenes and terpenoids across five contrasting urban greenspaces in Oxford, UK, and related chemical profiles to plant diversity, vegetation structure and surrounding habitat characteristics. Ambient air samples collected across spring and summer were analysed by thermal-desorption gas chromatography-mass spectrometry, alongside field surveys of taxonomic richness, canopy cover, vegetation volume and ground composition. Terpene and terpenoid scentscapes differed strongly among sites and through time, with monoterpenes dominating detected profiles. Sites with greater plant richness and greater vegetation volume below 1.5 m consistently showed higher terpene/terpenoid abundance, whereas total vegetation volume, canopy cover and tree abundance were weaker and less consistent predictors. These relationships were specific to plant-associated chemical classes and were not observed for predominantly anthropogenic volatile compounds. These patterns were resolved across only five independent sampling sites, and the relationships reported here should therefore be considered exploratory. Our findings suggest that human exposure to plant-derived volatiles in cities may be maximised not simply by increasing tree cover, but by designing biodiverse, structurally complex lower-to-mid-storey vegetation within the breathing zone. These results provide an empirical basis for incorporating scentscape ecology into health-oriented urban greenspace design.

Pesticide use is ubiquitous. While pesticide resistance in target species such as insects is a well-known phenomenon, it is less known how pesticide exposures affect the genetics of non-target species. Here, we demonstrate the research value of integrating existing datasets to study the relationship between chemicals and biodiversity from the genetics perspective by analyzing spatial pesticide concentration data and genetic data from Europe that were not initially generated for toxicological purposes. Specifically, we found associations between environmental pesticide concentrations and genetic variation (i.e., single nucleotide polymorphisms, SNPs) in a non-target species, the wild boar. Gene Ontology (GO) terms and biological signaling pathways related to nervous system function were overrepresented for the impacted genes, especially for insecticides and fungicides. Further, we found that external studies characterizing the molecular mechanisms of these pesticides revealed the same pesticide - GO relationships as those observed in our analysis. Our results show that pesticide-induced genetic changes may not be constrained to target species but may also have an impact on the long-term viability of other wildlife populations. We therefore encourage further research into this area and collaboration across disciplines to develop more comprehensive designated datasets for such analyses. This finding also urges the consideration and inclusion of chemical exposure into conservation protection strategies and greater consideration of genetic impacts from chemicals in environmental risk assessment.

Groundwater contamination by lead and nitrate exceeding the permissible limits of the World Health Organization (WHO) and the Bureau of Indian Standards (BIS) of 10 ppb lead and 45 to 50 ppm nitrate poses a severe health risk of systemic toxicity in humans and animals. Microbial nitrate reduction coupled with microbiologically induced calcite precipitation (MICP) proved to be a novel bioremediation strategy to transform soluble lead nitrate into insoluble lead biominerals. This study presents statistical optimization of operational parameters for nitrate reduction and co-precipitation of biogenic lead carbonate-calcium carbonate by nitrate-reducing bacterial consortia. The SEM, TEM with SEAD, and XRD analysis of the biomineral confirmed lead carbonate co-precipitation with CaCO3. A two-level factorial design was applied to screen significant variables and interactive factors that influence nitrate reduction and biomineral precipitation. The highest nitrate reduction, maximum ammonia production, and biogenic Pb-mineralization efficiency of > 90% were obtained at the optimum conditions (initial nitrate requirement of 600 ppm, temperature 37 oC, pH 8, and carbon content of 5.5 g/L) compared to those obtained under the common process parameters. ANOVA with a high correlation coefficient (R2 > 0.99) corroborated the linear model for the biogenic mineralization process. This study on the biogenic removal of Pb by nitrate-reducing bacterial consortia revealed possible opportunities for large-scale treatment of Pb-contaminated surface and subsurface water bodies.

Antimicrobial resistance (AMR) is a major threat to modern medicine and an emerging environmental pollutant. Wastewater-based monitoring offers a scalable alternative to traditional surveillance by enabling population-level monitoring across One Health compartments. However, it remains unclear how well bacterial variants detected in wastewater reflect those causing clinical infections. Here, we assessed the prevalence of carbapenem-resistant Escherichia coli (CREC) in wastewater from Uppsala, Sweden, and compared wastewater isolates with clinical isolates from the same region and period. We observed partial overlap between wastewater and clinical CREC populations, but several common clinical variants were not detected in wastewater and vice versa. CREC prevalence was an order of magnitude higher in wastewater receiving hospital effluent, yet this did not improve concordance with clinical isolates, suggesting that hospital wastewater may artificially inflate CREC prevalence estimates. These findings indicate that wastewater CREC prevalence does not necessarily reflect the regional clinical burden. Furthermore, metagenomic sequencing of wastewater-derived E. coli populations rarely detected CREC resistance genes, suggesting that CREC abundance is below the detection limit of this approach and that targeted enrichment is required for reliable monitoring. Together, our results show that while wastewater-based monitoring can complement AMR surveillance, it should be interpreted cautiously when used as an indicator of clinical risk.

The rapid global expansion of dams has significantly reshaped freshwater ecosystems, yet long-term trophic dynamics and food-web stability within reservoirs remain underexplored. This study examines the reorganization of fish communities induced by the trophic surge following dam construction and river impoundment. We used the predator-to-consumer mass ratio (PCMR) as a proxy for trophic pyramid structure to track changes in fish biomass distribution across three large boreal reservoirs in Quebec, Canada, over 20 years, using data from before and after impoundment. We found transient top-heaviness events (i.e., PCMR values > 1; predator accumulation), characterized by a hump-shaped trajectory of the PCMR values in two reservoirs (Opinaca and Robert Bourassa). The third reservoir (Caniapiscau) showed a gradual increase in top-heaviness but did not exhibit a clear transient pattern. The magnitude and duration of top-heaviness varied across reservoirs, potentially due to differences in the system's capacity to receive and accumulate nutrients, which are driven by water residence time and reservoir size. This research provides a conceptual framework for understanding the long-term trophic dynamics of reservoirs, which can help inform the development of more effective management strategies to preserve ecological health in these ecosystems.

A central question in ecology and evolution is determining when environmental challenges translate into stress and biological harm. Human-derived pollutants are often assumed to be inherently damaging; however, their effects may depend strongly on ecological and physiological context. Microplastics exemplify this complexity, eliciting heterogeneous responses across taxa rather than uniform toxicity. Here, we synthesise evidence from 14 meta-analyses encompassing 9,101 effect sizes to identify the conditions under which microplastic exposure leads to damage, neutrality, compensation, or stimulation. These syntheses reveal clear negative effects alongside frequent neutral outcomes and occasional stimulatory responses. Microplastic effects vary with particle characteristics and exposure conditions and are strongly moderated by species-specific traits and life stage. We further argue that plasticity in the gut and in behavioural and physiological traits may buffer microplastic effects. Emerging evidence also indicates that biofilm colonisation alters the physicochemical and biological properties of microplastics, potentially modifying their ecological interactions and toxicity profiles. By reframing microplastics as context-dependent challenges rather than universally toxic agents, we provide a conceptual framework for understanding animal responses under realistic ecological conditions and identifying the biological and environmental contexts in which microplastic exposure is most likely to result in harm.

Antibiotic resistance genes (ARGs) travel from cities into rivers and coastal waters through wastewater treatment plants. Monitoring them normally requires metagenomic sequencing, which is sufficiently costly that most utilities can sample only occasionally. Weather and location data are freely available on a daily basis for virtually any location worldwide, making them attractive predictors for identifying where limited sequencing resources should be prioritized. However, whether such models generalize to previously unseen cities remains largely untested, as most published studies train and evaluate models within the same catchments, thereby assessing interpolation rather than geographic transferability. To address this gap, we paired 235 wastewater metagenomes collected from five European cities with 23 abiotic predictors spanning geospatial, meteorological, hydrological, radiative, and temporal domains. Model performance was evaluated using leave-one-group-out (LOGO) cross-validation, in which all samples from one city were withheld for testing while the remaining cities were used for training. CatBoost achieved a median LOGO ROC-AUC of 0.929 and a median F1-score of 0.750. Using freely available environmental reanalysis predictors (meteorological, hydrological, radiative, geospatial, and temporal) - without any metagenomic sequencing - CatBoost achieved a mean ROC-AUC of 0.722, recovering 78% of the predictive performance of the full omics-integrated model. Removing latitude and longitude reduced ROC-AUC by only 0.003, whereas replacing random cross-validation with city-wise validation reduced ROC-AUC by 0.052. Predictive performance varied across ARG classes, ranging from a ROC-AUC of 0.981 for {beta}-lactam resistance genes to 0.762 for glycopeptide resistance genes. These findings demonstrate that freely available environmental reanalysis predictors - spanning meteorological, hydrological, radiative, geospatial, and temporal domains - recover 78% of the predictive signal for antibiotic resistance gene occurrence in urban wastewater, allowing scarce sequencing capacity to be directed to the catchments where it changes a decision.

BACKGROUND: Pesticides are extensively used in agriculture, raising concerns about their potential impact on human health through dietary and environmental exposure. OBJECTIVES: This study evaluated the in vitro cytotoxicity of ten commonly used pesticides and their mixtures (lambda-cyhalothrin, cypermethrin, deltamethrin, tebuconazole, glyphosate, acetamiprid, cyprodinil, piperonyl butoxide, fluopyram, and imazalil) on human intestinal Caco-2 cells. METHODS: Cytotoxicity was assessed using the MTT assay, as a measure of metabolic activity, and the trypan blue exclusion test, as an indicator of cell membrane integrity. FINDINGS: Results showed that high concentrations (100 mg/L) of all pesticides significantly reduced cell viability and vitality. Notably, glyphosate and tebuconazole exhibited significant toxicity even at lower concentrations, respectively 0.1 mg/L and 10 mg/L. Combination treatments (Top 3 and Top 8 pesticide mixtures) retained the cytotoxic effects observed for individual compounds, showing additive (non-synergistic) effects. CONCLUSIONS: Overall, these findings indicate that certain pesticides-based herbicides can exert cytotoxic effects on intestinal cells even at relatively low concentrations and highlight the importance of using the component-based approach in mixture risk assessment for humans. This study was performed as part of the EU SPRINT (Sustainable Plant Protection Transition: A Global Health Approach) project.

Wastewater-based epidemiology (WBE) has become a cornerstone of public health surveillance, yet routine regulatory monitoring remains largely focused on respiratory viruses and selecting antimicrobial resistant bacteria. Emerging foodborne parasites, such as Cyclospora cayetanensis, remain excluded from standard environmental surveillance panels despite increasing domestic outbreaks. This study presents a laboratory-based pilot protocol evaluating the feasibility of extracting and quantifying C. cayetanensis from urban wastewater across the Chicago metropolitan area. Using sample concentration via centrifugation and mechanical bead-beating, automated magnetic bead nucleic acid extraction, and digital PCR (dPCR), we demonstrate high-sensitivity detection in municipal influents and sewer nodes. These findings establish a proof-of-concept laboratory methodology to support the expansion of regulatory public health surveillance frameworks to include non-routine protozoan pathogens.

Wastewater-based epidemiology uses both targeted and non-targeted bioanalytical approaches to detect and track disease-causing microbes in wastewater. Targeted approaches such as quantitative polymerase chain reaction (qPCR) remain widely used while sequencing-based approaches have slowly gained popularity. However, concordance between established targeted molecular assays and sequencing approaches remains incompletely characterized. To assess the validity of sequencing as an alternative to qPCR, we compared outcomes of both methods targeting five respiratory viruses in wastewater samples collected across 16 Texas cities over 4 years (May 2022-July 2026) of continuous sampling. We found positive correlations between qPCR and sequencing measurements, with both methods independently correlating with statewide clinical emergency department visits. qPCR positivity also increased with genomic sequencing coverage, and sequencing demonstrated greater potential than qPCR as an early respiratory virus outbreak detection method. Our findings support metagenomic sequencing as a sensitive and highly specific alternative to targeted qPCR for wastewater surveillance, and reinforce its utility towards long-term longitudinal monitoring of respiratory viruses.

The contribution of environmental reservoirs to antimicrobial resistance (AMR) acquisition in humans remains poorly quantified, despite extensive evidence that resistant bacteria and resistance genes circulate through wastewater, surface waters, food and other environmental matrices. The central difficulty is no longer to demonstrate that these reservoirs are connected, but to quantify the direction and relative contribution of the pathways linking environmental AMR to human acquisition. Here, we synthesize the main processes that need to be represented at the interface between the environment and humans and propose a mechanistic framework for Escherichia coli producing extended spectrum beta lactamases (ESBL Ec), linking human, water and food compartments. The framework explicitly represents contamination of the environment by humans and multiple routes of human exposure to environmental reservoirs, including drinking water, recreational water exposure and food contamination mediated by irrigation. We apply the model to two contrasting illustrative settings, France and Madagascar, and use scenarios involving antibiotic use, environmental contamination and climate to explore how changes in specific drivers propagate across compartments. The simulations show responses that depend strongly on the compartment, while sensitivity analyses identify wastewater treatment efficiency and parameters describing transmission, exposure and environmental selection at specific interfaces as influential components of the system. The framework is used to explore how different transmission pathways and environmental drivers can shape AMR dynamics across interconnected human and environmental compartments. By comparing contrasting scenarios and identifying the parameters that most strongly influence model behaviour, we highlight both potential leverage points and critical gaps in current empirical knowledge. More broadly, this work illustrates how decomposing One Health AMR into tractable reservoir interfaces can help structure future data collection and progressively improve our ability to quantify transmission between reservoirs.

The decline in fishery yields in Japan's enclosed coastal seas has been accompanied by a reduction in phytoplankton biomass, suggesting a widespread decrease in ecosystem productivity. Oligotrophication in Mikawa Bay has caused substantial declines in phytoplankton biomass and Manila clam (Ruditapes philippinarum) production. To assess whether external nutrient enhancement could stimulate phytoplankton growth, a controlled demonstration experiment using treated effluent from two sewage treatment plants (WWTPs) was conducted (spanning fiscal years 2022 to 2024) based on analysis using a three-dimensional non-hydrostatic model. Although effluent DIP and nitrogen concentrations were increased, model results showed that WWTP-derived nutrients represent only a minor portion of the total nutrient budget. Field observations revealed limited increases in surface water phosphorus, strong seasonal variability, and only temporary rises in chlorophyll-a and clam harvests. Monthly TP data demonstrated that seasonal DIP pulses, especially spring phosphorus bursts, dominate nutrient dynamics and overshadow external loading effects. Laboratory experiments further showed that phosphate is efficiently fixed by Fe(III) under oxidizing conditions and rapidly released during reductive dissolution of [FeOOH-HPO4]2- complexes, confirming iron as the principal regulator of internal P cycling. Because current numerical models omit iron-driven internal cycling, they cannot accurately predict bioavailable P or ecosystem responses. Overall, the study indicates that localized nutrient enhancement is insufficient to counter long-term oligotrophication, and effective management must integrate nitrogen limitation, internal Fe and P cycling, oceanic exchange, and seasonal variability in Ise and Mikawa Bays.

Per- and polyfluoroalkyl substances (PFAS) are present throughout the environment due to decades of use in many commercial products and industrial processes. These molecules contain a hydrophobic fluorocarbon tail and a functional headgroup (typically a carboxylic or sulfonic acid) that allows hydrophilic interactions with the surrounding environment. These properties make them inherently resistant to degradation, difficult to track through the environment, readily bioaccumulative, and likely to impact human health. Through collaboration with town officials, multiple meetings with local sustainability committee members and school officials, and direct contact with individual homeowners, we conducted community-driven sampling of PFAS in well water from 105 houses in Tremont, ME, a rural coastal town that relies on private wells for drinking water. The rate of PFAS detection for one or more analytes across all wells (N=105) was 69%. Of all wells tested, 16 (15%) exceeded the current public drinking water standard in Maine (20 ng/L for 6 PFAS), and an additional 17 (totaling 31% of all wells tested) exceeded Maine's newly adopted public drinking water standards for the state (based on EPA 2024 standards). Because private wells are unregulated, homeowners may use these results as a guide for taking action to maintain safe drinking water. Both hierarchical cluster analyses and pairwise Spearman's rank correlation of PFAS test results revealed similar profiles in households near and far from one another. Along with the absence of obvious, large-scale potential point sources of PFAS contamination, these findings support septic systems as the primary source of PFAS contamination in drinking water across this town. The community-driven approach to the sampling effort has enabled information sharing on the importance of water filtration options, discussion of long-term pollution abatement strategies for individual households and the town, and focused efforts for future well water testing and PFAS mitigation.

Environmental exposure involves pollutant mixtures. Building on evidence that individual bisphenols alter zebrafish larval physiology and transcriptomic profiles at ng/L concentrations, we assessed the effects of specific mixtures of BPS, BPF, and BPAF without (M1) or with (M2) BPA. Experimentally, M1 and M2 induced 4.16% and 6.54% mortality in zebrafish larvae, despite the concentration addition model predicting no toxicity. Both mixtures reduced swimming-locomotor activities. These behaviors were accompanied by organ-level functional defects, as in vivo optical mapping revealed bradycardia and increased high-frequency (30-80 Hz) brain oscillations for both mixtures, while M2 additionally altered low-frequency (0.2-4 Hz) brain activity. The neurophysiological changes were accompanied by neurovascular remodeling, with in vivo two-photon microscopy revealing decreased vascular length and branching. Finally, transcriptomic analysis revealed common M1- and M2-induced alterations linked to pathways of vascular function, neuronal communication, and metabolism. M2 altered additional pathways of cardiac and neuronal signaling, consistent with the reported organ-physiology effects. We discuss the environmental relevance of mixture exposures and assess whether BPA intensifies mixture-induced responses.