PLOS Water

Background. Safely managed drinking-water monitoring under Sustainable Development Goal target 6.1 uses source-water quality, yet Escherichia coli is common in household drinking water in Bangladesh despite near-universal improved sources. We compared source and household point-of-use (POU) E. coli detection within households and estimated how the safely managed estimate changes if POU replaces source E. coli. Methods. Cross-sectional secondary analysis of the Bangladesh MICS 2025 water-quality subsample (February-June 2025; detection >=1 colony-forming unit/100 mL). We classified 5,944 households with readable paired results into four joint patterns and, exploratorily, examined POU detection among negative-source households and two-week diarrhoea in children under five with survey-weighted modified Poisson models. Results. E. coli was detected in 47.4% of source and 84.8% of POU samples; only 12.7% of households were negative at both. Among negative-source households, 75.8% had E. coli in household water, versus 94.7% with a detected source, persisting (33.3%) even direct from the source. The JMP-aligned safely managed estimate was 42.5% (95% CI 40.8-44.1); substituting POU for source E. coli gave 12.6% (11.4-13.8), not an SDG indicator estimate. The diarrhoea prevalence ratio for POU detection (1.04, 0.59-1.86) was too imprecise to interpret. In exploratory spatial analysis, POU detection clustered geographically (Moran's I 0.16 cluster level, 0.31 district level, both p<0.001) and was independently associated with lower elevation (odds ratio 0.62, 0.56-0.70) and longer travel time to healthcare (1.65, 1.29-2.12); five districts had complete detection among 84-94 households tested. Conclusions. Source and POU classifications differed substantially; the survey cannot show where or when contamination occurred. Contamination was spatially clustered, arguing against pure noise and pointing to districts for follow-up. The findings support evaluating standardised POU sampling as a complement to source-based monitoring, with targeted follow-up and cross-season, cross-country replication.

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.

Coastal waters often experience fecal contamination from human (e.g., septic systems, wastewater treatment catchments and outfalls) and non-human (e.g., wildlife, domesticated pets, and livestock) origins, posing human health risks through contaminated fisheries and recreational waters. Quantitative PCR (qPCR) methods can identify and quantify fecal origins for their management by targeting Bacteroides, a dominant group of gastrointestinal bacteria with high host specificity. This study identified major contributors and drivers of fecal contamination to Mobile Bay, Alabama, an urbanized estuary in the southern United States. We utilized a multi-species qPCR source tracking approach targeting total Bacteroides and human-, pig-, dog-, cow-, and fowl-associated Bacteroides. Water samples and environmental parameters were collected over one year from 13 sites that represent a range of potential fecal inputs common to coastal waters. Human-associated feces were a consistent major contributor to the system (positive in 82% of samples), with localized inputs from pig (wild or domestic) and dog. Freshwater discharge and nitrogen constituents were good predictors of Bacteroides concentrations in this system, while areas with impervious surfaces and developed land cover may be contributing elevated fecal indicator levels. The multi-species qPCR source tracking approach described in this study can be implemented in other watersheds to support the development of better detection, identification, and fecal management strategies for community health of coastal areas.

This study developed a streamlined, sensitive and rapid method for co-detection of low concentrations of Salmonella enterica, Shigella flexneri, Listeria monocytogenes, and pathogenic Escherichia coli from soil. First, autoclaved soil from Iowa with serial dilutions of bacteria to confirm individual species recovery rates on selective agar by qPCR under varying primary enrichment times was used. All pathogens were detected by qPCR of colonies recovered from replicate sterile soils spiked with 10^1 or greater bacteria within 5 hours of enrichment. Shiga toxin expressing E. coli (STEC) and L. monocytogenes were consistently co-recovered in autoclaved and natural soil, pathogen spiking concentration, and secondary enrichment time. S.enterica was consistently recovered at autoclaved and natural concentrations, while S. flexneri detection rates fluctuated. Twenty-three soil samples from Kenyan neighborhoods, with prior evidence of contamination, was compared to qPCR detection at each protocol stage: processing, primary enrichment, secondary enrichment, and selective colony isolation. Detection rates substantially increased for S. enterica, STEC and other pathogenic E. coli after enrichment, with optimal detection rates occurring after secondary enrichment. Testing colonies from selective agar resulted in high false negative rates, possibly reflecting competition from native soil microbiota that was absent in spiked sterile soil experiment. Evaluation of the method with other environmental sample types yielded consistent detection patterns. Evidence suggests a protocol including 24-hour primary enrichment, followed by 24-hour secondary enrichment, and qPCR testing of soil DNA at primary and secondary steps is a reproducible and more expedient method for co-detecting common Enterobacteriaceae and L. monocytogenes pathogens in soil.

Background: In coastal Bangladesh, saline drinking water has been linked to raised blood pressure. The 2025 Multiple Indicator Cluster Survey (MICS) measured electrical conductivity (EC), a salinity indicator, in source water, but its report gave no population counts or district estimates. Objectives: To estimate how many people live in households whose tested source has high EC, and to describe patterns by place, household characteristics, source type and arsenic. Methods. EC was measured in source water of 6,040 households in 64 districts, February to June. In 14.5% of households the tested source was not the main drinking-water source. Primary outcomes were EC above 1,500 and 3,000 microsiemen/cm, survey risk classes that are not health-based standards. Estimates were survey weighted; comparisons were observational. Results: EC exceeded 1,500 microsiemen/cm in 5.3% of households (95% CI 4.7 to 5.9), about 9.0 million people, including 1.8 million above 3,000 microsiemen/cm. Coastal-zone districts held 26% of households but 70% of those above 1,500 microsiemen/cm. Shares varied little by wealth or in households with young children or pregnant women. Nationally, deep tube wells more often had high EC because most are coastal. In the coastal zone, deep and shallow wells had similar shares (16.1% and 17.9%). After district adjustment, EC was about 18% lower in deep wells (geometric mean ratio 0.82, 0.73 to 0.92). Within upazilas (restricted sample) it was 0.93 (0.80 to 1.07). Arsenic above 10 ppb was more common where EC was high, but this link weakened after district adjustment. Conclusions: Millions of people live in households whose tested source has high EC, mostly in coastal and south-central districts. Deep wells had modestly lower EC after geographic adjustment, but the difference shrank within upazilas and depended on the handling of implausibly low readings. The findings support measuring EC alongside arsenic, with documented meter calibration.

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.

Biological filtration in newly established aquaria depends on the establishment of nitrifying microorganisms that prevent accumulation of toxic ammonia and nitrite. Many aquarists use commercial nitrifying supplements to accelerate biofilter establishment and reduce ammonia and nitrite accumulation. However, the microbial composition and in situ efficacy of these products remain poorly characterized. We evaluated the ability of enrichment cultures containing comammox Nitrospira (CX) and the ammonia-oxidizing archaeon Ca. Nitrosotenuis aquarius (AQ) to serve as aquarium nitrifying supplements, tested alongside a conventional supplement (CS) containing ammonia-oxidizing bacteria. Inoculation of new aquaria containing zebrafish (Danio rerio) with nitrifying supplements resulted in distinct nitrifier profiles within biofilters of each treatment group corresponding with significant reductions in peak ammonia and nitrite concentrations, which were most evident when applied at the maximum tested supplement dose. Treating aquaria with CX, AQ, or CS maintained ammonia and nitrite at low concentrations, and CX effectively prevented ammonia and nitrite accumulation through all stages of aquarium operation and biofilter establishment. After removing all fish from the aquaria, ammonia "spike-in" experiments demonstrated biofilters treated with CX and AQ conferred long-term protection against nitrite accumulation following acute ammonia loading events. Whereas 16S rRNA gene sequences associated with CS were rarely detected within biofilter profiles, those associated with CX and AQ established early and persisted temporally. Overall, results demonstrate how nitrifiers might be differentially adapted to home aquarium conditions and the potential of AOA and comammox Nitrospira to improve biofilter establishment and maintaining safe water quality in newly established freshwater aquaria.

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.

Background: Chronic arsenic exposure in drinking water causes skin lesions and cancers and raises infant, cardiovascular and all-cause mortality in Bangladesh. National surveys tested source water in 2012-13, 2019 and 2025 with different kits, so the trend is uncertain. Objectives: We estimated how arsenic in source water changed after laboratory adjustment of 2012-13 readings, and how many people remained exposed. We also assessed whether districts high in earlier surveys held most people exposed in 2025. Methods: We analysed 11,624 households in three Multiple Indicator Cluster Surveys. We counted arsenic strictly above 10 and 50 parts per billion (ppb). Results: Estimates reproduced published 2019 and 2025 tables. After laboratory adjustment of 2012-13, the share above 10 ppb fell from 24.0% to 14.4% (adjusted ratio 0.61, sampling 95% CI 0.55 to 0.67; 0.69 with interview month). Above 50 ppb it fell from 10.9% to 8.6% (0.80, 0.69 to 0.93; 0.93 with month). Most of the fall at 10 ppb occurred within source types. Without false positives, it would vanish only if later kits detected 61% of sources above 10 ppb. About 26 million people used such water in 2025 (kit scenarios 23 to 32 million). The 25 districts high in 2012-13 and 2019 held 39% of the population but 72% of those exposed (bootstrap 95% interval 60 to 76). Clustering was evident, but no district was a significant hotspot. Conclusions: Arsenic above 10 ppb was lower in 2025 than in 2012-13, but the size of the fall depends on unverified later kits. Change at 50 ppb and since 2019 was unclear after month adjustment. One household in seven exceeded the WHO guideline in 2025. Districts found high earlier held most exposed people, supporting prioritised well testing there alongside broader surveillance. Future surveys need one kit with a laboratory cross-check.

Population-scale pathogen surveillance is limited worldwide, particularly in resource-constrained settings, where clinical systems only monitor select priority pathogens and reach only those who can access healthcare. Wastewater-based epidemiology addresses many of these gaps, offering cost-effective surveillance that is adaptable across diverse implementation contexts. In Lagos, Nigeria, a high enteric pathogen burden coincides with a complex sanitation landscape. However, circulating pathogen diversity in Lagos is largely uncharacterised outside of outbreak contexts. Here we characterize human-associated pathogen diversity in Lagos with virus-enriched metagenomic and metatranscriptomic sequencing of untreated wastewater sampled from open drainage canals from July-August 2024. We detected a diverse human-associated virome dominated by enteric adenoviruses, astroviruses and caliciviruses. We recovered numerous partial and near-complete genomes from pathogens of public health concern, including noroviruses, enteroviruses and sapoviruses. Although our enrichment targeted viruses, we detected diverse bacterial pathogens, including recurrent detection of Vibrio cholerae, other enteric bacteria, and zoonotic pathogens such as Streptococcus suis. We also detected antimicrobial resistance genes, including aminoglycoside, beta-lactam and fluoroquinolone resistance and clinically important genes such as mcr. Together, these results establish a regional baseline of pathogen diversity and demonstrate the utility of virus-enriched sequencing for integrated wastewater surveillance.

Background: Lead is a cumulative environmental toxicant for which no safe level of exposure has been identified, and pregnant women and their fetuses are particularly vulnerable because lead readily crosses the placenta and can be mobilised from maternal bone stores. Bangladesh conducted its first nationally representative blood lead testing among pregnant women through the 2025 Multiple Indicator Cluster Survey (MICS), but a detailed, multi-domain analysis of the determinants of elevated blood lead level (BLL) among this population has not yet been reported. Methods: We conducted a secondary cross-sectional analysis of 1,928 pregnant women aged 15-49 years with complete data from the women's dataset of Bangladesh MICS 2025. Elevated BLL was defined as [&ge;]5 g/dL. Survey-weighted logistic regression was used to estimate unadjusted and adjusted odds ratios (aOR) for co-exposure to arsenic, cadmium and mercury and sociodemographic and socioeconomic covariates. Slope Index of Inequality (SII) as well as Relative Index of Inequality (RII) were used to quantify wealth-related inequality. Population attributable fractions (PAF) were estimated for three counterfactual scenarios, and adaptive least absolute shrinkage and selection operator (LASSO) regression was used as a sensitivity analysis. Results: Elevated BLL was present among 6.2% of participants. In the adjusted model, a 2-fold rise in blood cadmium was associated with higher odds of elevated BLL (aOR: 1.54; 95% CI: 1.21-1.96), whereas a 2-fold rise in blood mercury (aOR: 0.63; 95% CI: 0.48-0.83), rural residence (aOR: 0.55; 95% CI: 0.32-0.94), secondary education (aOR: 0.49; 95% CI: 0.28-0.85) and post-secondary education (aOR: 0.22; 95% CI: 0.11-0.46) were associated with lower odds. Belonging to progressively wealthier quintiles was associated with markedly higher odds, BLL rising to an aOR of 10.32 (95% CI: 4.41-24.15) for the richest versus poorest quintile. The RII was 8.59 (95% CI: 4.24-17.40) and the SII was 0.159 (95% CI: 0.101-0.218), indicating a substantial pro-rich gradient in elevated BLL. Counterfactual scenarios suggested that shifting the population to secondary-level education could reduce the prevalence of elevated BLL by 52.9% (95% CI: 20.6-72.0) and shifting to rural residence by 24.3% (95% CI: 1.4-41.9). Adaptive LASSO variable selection produced estimates consistent in direction and magnitude with the fully adjusted model. Conclusion: In this national sample, approximately one out of every 16 pregnant women had blood lead levels that met or exceeded the action threshold set by the World Health Organization, with this issue being particularly prevalent among wealthier, urban, and less-educated women, and was compounded by co-exposure to cadmium. In Bangladesh, the typical pattern observed for maternal and child health indicators does not align with the inverse relationship found between household wealth and lead exposure. This suggests that the sources of exposure are not merely indicators of material poverty. Antenatal blood lead screening, source investigation, and equity-informed lead-reduction strategies that specifically reach wealthier and urban households are warranted.

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.

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.

Okanagan sockeye salmon have shown a strong recovery over the past three decades, but the contribution of environmental conditions and management actions to this recovery has not been determined. We developed a hierarchical Bayesian model linking spawner abundances to fry emergence observations to evaluate how flow and water temperature during adult upstream migration and incubation periods influence spawner-to-fry production. Production declined by 62% in years with earlier onset and longer duration of periods when warm river temperatures created barriers to adult migration, indicating that migration conditions can have carry-over effects on reproductive success even when spawners successfully reach the spawning grounds. Additionally, production was 46% lower when flows fell above or below the management target range designed to minimize scour and desiccation during egg incubation. In contrast, we found little support for effects of pre-spawning flows or incubation temperature on production. These findings provide quantitative evidence that current flow management is increasing freshwater spawner-to-fry production and has likely contributed to the Okanagan sockeye recovery. Effective flow management may therefore help offset climate-driven impacts that increasingly threaten this population.

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.

Freshwater biodiversity is declining at alarming rates globally, yet our capacity to monitor it depends heavily on the methods used for biodiversity assessment. Stream benthic macroinvertebrates are widely used as bioindicators, but traditional morphological identification is labor-intensive and limited by taxonomic expertise, while DNA metabarcoding offers a promising but incompletely evaluated alternative. Here, we compared biodiversity patterns detected by four macroinvertebrate sampling and identification methods (morphological identification of benthic samples, morphological identification of combined benthic and stream-edge samples, eDNA metabarcoding of bulk benthic samples, and eDNA metabarcoding of streamwater) across 62 sites in seven watersheds spanning the southern continental United States. We compared methods across three facets of biodiversity (gamma diversity, alpha diversity, and taxonomic composition) and three taxonomic levels (family, genus, and species). We found that which method detected more taxa shifted with taxonomic resolution: morphological methods detected more families and genera, while metabarcoding methods detected more species. Bulk benthic metabarcoding detected more families and genera than water eDNA metabarcoding, consistent with the "watered-down biodiversity" effect of DNA dilution in water samples, but the two metabarcoding methods did not differ significantly in species richness. All four methods detected largely distinct suites of taxa, with metabarcoding methods yielding more unique taxa than morphological methods. Notably, taxa uniquely detected by water eDNA were dominated by soft-bodied organisms (oligochaetes, leeches, earthworms) and water-column-associated taxa, while benthic eDNA uniquely detected hard-bodied EPT insects. We attribute this pattern to fundamental differences in eDNA shedding, transport, and deposition dynamics across organism types. These results demonstrate that no single method captures a complete picture of stream macroinvertebrate biodiversity, and that method choice should be guided by the taxonomic resolution and community components most relevant to the goals of the bioassessment program.

Hydropower development has emerged as a major driver of global environmental change, causing significant ecological damage to freshwater ecosystems and threatening aquatic biodiversity. However, the impact of cascade dams on fish communities in large river systems remains poorly understood, particularly in biodiversity hotspots such as China. Here, we used environmental DNA (eDNA) metabarcoding to assess the spatiotemporal patterns of fish diversity and community structure in the Ganjiang River, the largest tributary of the Yangtze River, in relation to cascade hydropower development. We collected water samples from 19 sites along the main stem and major tributaries in both wet and dry seasons, and analyzed the eDNA data using high-throughput sequencing and advanced bioinformatics pipelines. Our results revealed significant seasonal and spatial variations in fish diversity and community composition, with lower species richness and distinct assemblages in the impounded sections upstream of the dams compared to the downstream reaches. The fish communities were strongly influenced by environmental factors such as water temperature, pH, dissolved oxygen, and nutrients, which exhibited complex interactions with dam operation and land use practices. Spatial scale analyses further indicated that the impact of dams on fish diversity was most pronounced at the local scale, but also detectable at the regional scale. Our findings provide novel insights into the ecological consequences of cascade hydro-power development on fish communities and highlight the importance of considering spatial scales and environmental heterogeneity in assessing the impacts of human activities on freshwater biodiversity. Our study demonstrates the power of eDNA metabarcoding as a cost-effective and non-invasive tool for biodiversity assessment and conservation planning in large river systems facing multiple stressors and knowledge gaps.

Objective: To evaluate the specimen-level performance of a multichannel electronic nose (e-nose) sensor array in classifying stool specimens into relevant Clostridioides difficile categories. Design: Retrospective single-center validation study with reference to polymerase chain reaction (PCR) and C. difficile toxin testing. Participants: A convenience sample of 31 discarded stool specimens with PCR and confirmatory toxin testing. Categorized into PCR-positive (+)/toxin+, PCR+/toxin-negative (-), and PCR-. Methods: Specimens were sampled in controlled headspace by two e-nose sensor arrays in 10-minute sessions with 5-second sampling intervals. The primary endpoint was one-versus rest classification performance (sensitivity, specificity, positive predictive value, and accuracy) into three classes: PCR+/toxin+, PCR+/toxin-, and PCR-. Data were normalized and the region of interest was extracted for training and testing. Random forests were utilized for classification of normalized data with a test-train split at the specimen-level. Results: There were 457 unique 10-minute sampling events (147 PCR+/toxin+, 183 PCR+/toxin-, 127 PCR-). Sensors 1 and 2 captured 20,331 and 23,538 unique 5-second exposures within the region of interest, respectively. Testing data classification accuracy was 0.79 [0.78, 0.81] and 0.84 [0.83, 0.85] for Sensors 1 and 2. Sensitivity for PCR+/toxin+ exposures was 0.82 [0.79, 0.84] and 0.94 [0.92, 0.95], and specificity was 0.91[0.90, 0.93] and 0.93 [0.92, 0.94] for Sensors 1 and 2. Conclusion: An e-nose sensor array partially differentiated PCR-classified C. difficile specimen classes at the exposure level under headspace-controlled conditions. Performance was strongest for PCR+/toxin+ exposures. Findings support further prospective validation in the hospital room setting.

Background: Childhood lead exposure is a major global health concern. The 2025 Bangladesh Multiple Indicator Cluster Survey (MICS7) measured blood lead in a nationally representative sample of children aged 12-59 months for the first time. Preliminary reports suggested elevated blood lead was more common in wealthier urban households, contrary to the usual assumption. Methods: We analysed 10,622 children with a valid blood lead result (10,458 with complete covariate data for regression models). Elevated blood lead was defined as >=5 ug/dL. Prevalences are survey-weighted with design-based confidence intervals. Associations were estimated with survey-weighted logistic regression using stratified-cluster (Taylor-linearised) variance, and checked with a stratified bootstrap of primary sampling units and Benjamini-Hochberg false-discovery-rate control. Sensitivity analyses varied the outcome definition, the sample, and the weighting. Results: Survey-weighted prevalence of elevated blood lead was 38.5% (95% confidence interval [CI] 37.2-39.9%). Prevalence rose monotonically from 30.2% in the poorest to 51.9% in the richest wealth quintile. In the fully adjusted model, each wealth quintile was associated with 23% higher odds (adjusted odds ratio [AOR] 1.23, 95% CI 1.16-1.30), each level of maternal education with 20% lower odds (AOR 0.80, 95% CI 0.76-0.84), and urban residence with 37% higher odds (AOR 1.37, 95% CI 1.18-1.59). Household crowding was associated with higher odds (AOR 1.17 per person per room, 95% CI 1.10-1.24). The wealth-education interaction had AOR 0.95 (95% CI 0.92-0.98). In an exploratory contrast, weighted prevalence was 76.2% among children in the richest quintile with mothers having at most primary education versus 19.5% among those in the poorest quintile with mothers having higher secondary or tertiary education (prevalence ratio 3.9, 95% CI 2.6-6.0). Adjusted odds relative to Barishal ranged from 0.39 in Rajshahi to 3.29 in Dhaka. Conclusions: In this cross-sectional survey, elevated blood lead was more prevalent among wealthier and urban households and among children of less educated mothers, with large geographic variation. Lead prevention and source-identification efforts should therefore not be restricted to poor communities; prevalence remained high even in the poorest quintile. Interpretation is limited by substantial missing blood lead results and coding uncertainty.

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.