PLOS Water

Since the COVID-19 pandemic, wastewater-based surveillance (WBS) has emerged as a key approach to assess community-level health and the evolution of pathogens. To date, most established WBS systems focus on polymerase chain reaction (PCR) based detection and targeted sequencing of known pathogens because these approaches are well-accepted and include amplification of pathogen target sequences of interest thereby enabling lower limits of detection. Metagenomic next-generation sequencing (mNGS) is a promising approach to enable pathogen detection and surveillance beyond predefined pathogen lists, but its regular application to WBS has not been yet widely adopted because many key performance characteristics are not well-understood, including limit of detection (LOD) and false positive/negative rates. This paper describes a computational analysis to estimate the operational LOD of various sequencing approaches using a simplified model of a local wastewater (WW) system involving a military base. This paper also presents findings from two types of experiments: 1) laboratory-spiked, those for which Atlantibacter subterraneus (Asub) is introduced into real-world WW samples in a laboratory setting, and 2) system-spiked, those for which Asub is introduced at a source location of a real-world WW system. Findings indicate that mNGS detection performance varies with sequencing method and the data analysis process. In addition, findings indicate that site-specific method characterization should be used when implementing mNGS for WBS because sites can have different WW system configurations, background organisms and sequencing inhibitors.

Background: Population-based biomonitoring of contemporary-use pesticides remains limited in the United States, particularly in rural agricultural regions, and few studies have repeated measurements within the same individuals over time. Methods: We analyzed 28 urinary pesticide-related biomarkers among 600 adults from the population-based Survey of the Health of Wisconsin with archived urine collected during 2008-2016; 296 participants provided repeat urine and updated exposure information in 2025. Detection frequencies, co-detection, and within-person detection patterns were characterized. Generalized estimating equations were used for stacked, repeated-measures analyses of factors associated with detection of aminomethylphosphonic acid (AMPA), glyphosate, 2,4-dichlorophenoxyacetic acid (2,4-D), and any of these three. Prospective-only analyses evaluated more detailed agricultural and recent exposure measures. Results: Glyphosate, AMPA, and 2,4-D were detected in 7.7%, 6.2%, and 4.3% of retrospective specimens and 5.4%, 3.1%, and 4.1% of prospective specimens, respectively. Co-detection and persistent detection across the 9 to 17-year interval was rare. In repeated-measures models, greater fruit and vegetable intake, older age, and male sex were associated with higher 2,4-D detection. Lower household income was associated with lower AMPA detection, while afternoon/evening collection was associated with higher AMPA detection. In prospective analyses, working on field-crop agricultural land showed the strongest agricultural associations, particularly for 2,4-D and detection of any of the three pesticides. Associations were not seen with self-reported conventional versus organic produce consumption. Conclusions: Urinary pesticide detections were generally infrequent in this Wisconsin population. Diet and direct agricultural activities may be more informative exposure pathways than residing near cropland or private well drinking-water characteristics.

Summary Background: Wastewater and environmental surveillance (WES) enables community-level monitoring of infectious diseases. Most progress has focused on sewer-based surveillance, yet nearly half of the global population relies on non-sewered sanitation. In non-sewered settings, urban drainage channels have been used for poliovirus environmental surveillance, but their potential for a multi-pathogen WES with spatially defined catchments, and comparability to sewer-based surveillance, remains under-explored. Methods: Ten drainage channel sampling points with delineated micro-catchments (0.95-3.83 km2 with 12,885-44,146 people) were selected within Kampala. A total of 255 drainage channel and 54 wastewater treatment plant (WWTP) influent samples were collected during two campaigns in March and September-October 2025. A multi-target panel was quantified by digital PCR, including enteric viruses (Norovirus GI and GII, Rotavirus), respiratory viruses (SARS-CoV-2, Influenza A and B viruses, Respiratory Syncytial Virus (RSV)), non-O1/O139 Vibrio cholerae (V. cholerae), and Pepper Mild Mottle Virus (PMMoV) as a fecal indicator. Findings: Enteric viruses, non-O1/O139 V. cholerae, and PMMoV were consistently detected across all drainage channels and WWTP influents. Concentrations were generally lower in drainage than WWTP influents, except for non-O1/O139 V. cholerae. When normalized by PMMoV, concentrations across most drainage channels were comparable to WWTP influents, although comparability varied by target and location. Both concentrations and PMMoV-normalized concentrations varied across sampling locations. Trends between campaigns varied by pathogen target and were not explained by any single micro-catchment characteristic. Influenza A virus was the most frequently detected respiratory virus (8-23% in drainage channels; 3-10% in WWTP influents), while SARS-CoV-2, Influenza B, and RSV were rarely detected. Interpretation: PMMoV-normalized concentrations across most micro-catchments were comparable to WWTP influents, with spatial heterogeneity implying neighborhood-level differences in disease prevalence. Catchment-delineated drainage surveillance has potential to offer spatially resolved public health information in non-sewered settings comparable to sewer-based wastewater monitoring. Funding: Eawag Discretionary Funding

Freshwater ecosystems in Scotland are increasingly threatened by multiple interacting stressors, including pollution, hydrological alteration, land use change, and climate-driven pressures. Scotland's rivers are amongst the most physically diverse and dynamic in the UK, contributing significantly to the country-s economy and natural heritage. Monitoring 125,000 km of waterways presents an issue, with limited funding and capacity of environmental agencies creating an environmental monitoring deficit at a time when robust data are essential for delivering national and global biodiversity targets. Citizen science offers a scalable, community driven approach to strengthening environmental evidence. This study evaluates the development, implementation, and outcomes of Guardians of our Rivers (GooR), a nationwide citizen science programme using macroinvertebrate-based river health assessments based on the Anglers' Riverfly Monitoring Initiative (RMI) established by the Riverfly Partnership. Between 2022 and 2025, GooR trained more than 850 volunteers, established 123 monitoring sites across Scotland, and generated over 860 surveys-surpassing the previous 16 years of aquatic invertebrate monitoring efforts. Standardised field training, full equipment provision, and structured support enabled high-quality data collection across diverse catchments. The establishment of nationally applied trigger and target thresholds created a consistent mechanism for detecting ecological deterioration and ensured that verified trigger breaches (n = 39 in 2025) resulted in formal investigation or follow-up. Analysis of national abundance patterns revealed ecological and biogeographic trends across the eight RMI taxa, confirming data sensitivity to habitat differences, water quality, and regional pressures. A detailed case study of the Lothian Esk catchment demonstrated the programme's capacity to detect seasonal macroinvertebrate dynamics and site level ecological trajectories over time. Overall, GooR illustrates how well-designed and well-supported citizen science can deliver high resolution spatiotemporal datasets, enhance early warning systems, empower communities, and make a meaningful contribution to promoting river health. Importantly, GooR forged a working partnership with Scottish Environmental Protection Agency (SEPA) that allowed communities to not only gather data but also transformed the way citizen science is recognized as contributing to freshwater conservation. Continued investment and long-term support will be essential to sustain these gains and realise the full potential of citizen-driven freshwater stewardship.

Wastewater-based epidemiology (WBE) has emerged as a powerful population-level surveillance tool, but its coverage is structurally concentrated in in-network urban areas, potentially leaving rural populations underrepresented. Routine human movement between sewered (in-network) and unsewered (off-network) areas may, however, cause wastewater treatment plant (WWTP) measurements to reflect infectious disease dynamics beyond sewer boundaries. We evaluated this hypothesis using daily clinical COVID-19 testing data (January 2021-April 2022) across New York State excluding New York City (NYC). We disaggregated weekly cases and tests into in-network (WWTP catchment area) and off-network (outside WWTP catchment area) components applied to two geographic frameworks: administrative counties (N = 53 mixed-coverage) and mobility-defined communities identified through Walktrap community detection applied to census tract-level movement networks (N = 32 mixed-coverage). In/off-network COVID-19 trends were strongly correlated under both frameworks. County-level statewide aggregate correlations were high (incidence r = 0.994, positivity r = 0.996), as were individual county correlations (median r = 0.909 and 0.932, respectively). Mobility-defined community-level statewide correlations were similarly strong (r = 0.990 and 0.992), with comparable unit-level medians (r = 0.877 and 0.894). The mobility-defined community framework provided better population balance between in-network and off-network strata (87.5% vs. 69.8% in balanced range) and a higher floor on representativeness (minimum r = 0.440 vs. 0.177). Population size was the dominant predictor of in-network/off-network alignment at both scales; wastewater infrastructure density and off-network signal variability provided additional explanatory power at the mobility-defined community level. WWTPs broadly represent COVID-19 dynamics in surrounding off-network populations, supporting their use as sentinel surveillance sites. Representativeness weakens in smaller, more rural communities, and mobility-defined communities provide a complementary framework for identifying where this occurs.

Karst groundwater supplies in Virginia are relied on to varying degrees for domestic, agricultural, and municipal water supplies. Further, Virginian caves harbor an estimated 200 endemic invertebrate species. The microbial occupants of Virginias karst aquifers are largely undescribed; characterizing them promises to inform both the scientific description of these systems and the management of a critical water resource. Karst aquifers are heterogeneous, and much of the water moving through them cannot be reached directly; we profiled cave waters both because cave passages offer direct access to active groundwater and because cave water specifically is relied upon by endemic invertebrate species living in caves. Using 16S rRNA sequencing, we characterized aquatic microbial communities in eight Virginia caves, across Virginias four major karst regions. We identified 3,899 unique amplicon sequence variants (ASVs) and found that caves hosted diverse microbial assemblages that differed markedly among sampled sites. These baseline data provide a starting point for future work to understand how seasonal cycles, weather events, and surface disturbances affect the microbial communities present in cave waters and the cave-endemic invertebrates that depend on these waters.

Wastewater systems are increasingly recognized as important environmental reservoirs of antimicrobial resistance (AMR), acting as interfaces where resistant bacteria, antimicrobial resistance genes (ARGs), and mobile genetic elements (MGEs) converge and potentially disseminate. Wastewater samples were collected from hospital and community sites, including municipal medical centers, household wastewater outlets, and open drainage systems. Genomic DNA was extracted using the ZymoBIOMICS DNA/RNA Miniprep Kit and sequenced on the Oxford Nanopore Technologies MinION MK1D platform using the Native Barcoding Kit (SQK-NBD114.24, V14). Sequencing data were processed through a custom Snakemake workflow integrating quality control, taxonomic profiling, resistome characterization, mobilome analysis, and genome-resolved metagenomics. A total of 489 unique ARGs conferring resistance to 29 antibiotic classes were identified through metagenomic analysis. The resistome was dominated by genes conferring resistance to {beta}-lactams (including cephalosporins and carbapenems), aminoglycosides, tetracyclines, macrolides, and fluoroquinolones. Clinically important resistance determinants, including blaOXA, blaTEM, blaGES, blaCARB, cfxA, tet, qnr, sul, dfrA, erm, msrE, and aminoglycoside-modifying enzyme genes such as aac(3) and ant(3'') were detected across both hospital and community wastewater samples. Resistance mechanisms were predominantly driven by antibiotic inactivation, followed by efflux and target protection. Several priority bacterial pathogens were detected, including Escherichia coli, Klebsiella pneumoniae, Enterobacter cloacae, Pseudomonas aeruginosa, and Acinetobacter baumannii. Integration/excision elements were the predominant category of MGEs, followed by transfer-associated elements and replication/recombination/repair functions. Plasmid analysis further identified diverse incompatibility groups, predominantly IncP6, IncC, IncF, and IncR replicons, supporting the widespread occurrence of plasmid-mediated horizontal gene transfer in both settings. These findings reveal a substantial burden of clinically relevant ARGs, mobile genetic elements, and potential bacterial pathogens in hospital and community wastewater in Conakry. This study provides the first metagenomic baseline for environmental AMR surveillance in Guinea and highlights the urgent need for integrated One Health strategies to mitigate the environmental dissemination of antimicrobial resistance.

Newly established freshwater aquaria rely on development of biofilter nitrifying populations to prevent ammonia and nitrite accumulation that can negatively impact fish health. Although initial fish loads impact water chemistry of new aquaria, little is known about the corresponding impact on microbial community succession within freshwater aquarium biofilters. To address this gap, fourteen home aquarium systems were established, stocked with a range of fish loads, and maintained for eight months. Aquaria were sampled regularly to monitor nitrogen species, microbial community composition (16S rRNA gene sequencing), and the abundance of nitrifiers (qPCR). Aquaria with higher fish loads developed microbial communities that were compositionally distinct from those with lower fish loads, and were dominated by Pseudomonas, Rhodobacter, and Planctomycetes. These patterns are consistent with increased nutrient availability supporting biofilm development, whereas lower fish loads may delay biofilm maturation. Increasing the number of fish in an aquarium significantly increased maximum ammonia and nitrite concentrations, although both were ultimately depleted within similar timeframes across treatments. Comammox Nitrospira were among the most abundant biofilter nitrifiers and were present in all biofilter samples regardless of fish load. Ammonia-oxidizing bacteria were detected at relatively low abundance but showed increases in relative abundance within high fish load aquarium filters. Ammonia-oxidizing archaea were below sequencing detection limits and detected only at low levels by qPCR, suggesting that their establishment in aquarium biofilters may require higher initial inoculation or longer timeframes. Overall, these results demonstrate that fish load shapes microbial community development in newly established aquarium biofilters, and that comammox Nitrospira dominate among nitrifiers during early biofilter establishment.

Freshwater bioassessment relies on assessing aquatic assemblages to infer ecological conditions, yet conventional surveys require extensive field sampling, specimen processing, and specialized taxonomic expertise. Existing environmental DNA (eDNA) methods have not yet provided a practical alternative to conventional macroinvertebrate assays in part because current approaches cannot feasibly recover broad taxonomic diversity at sufficient taxonomic resolution. Here, we evaluated targeted hybridization capture of mitochondrial cytochrome oxidase I (COI) target sequences as a unified molecular approach for cross-phylum freshwater bioassessment. Environmental DNA was collected at 18 sites along 63 km of Boulder Creek spanning nearly 1,500 m of elevation from forested headwaters to agricultural plains. COI targets were enriched using custom RNA bait panels designed to target regional freshwater arthropods, annelids, and molluscs. Hybridization capture increased recovery of COI sequences [~]1,760-fold relative to unenriched shotgun libraries, generating Folmer-region COI contigs that averaged [~]400 bp. Across the watershed, we recovered sequences for approximately 450 macroinvertebrate genera across 8 phyla. Detected macroinvertebrate richness averaged 56 genera per site and increased down Boulder Canyon before declining downstream of the city. Macroinvertebrate assemblage composition from hybridization capture paralleled patterns observed with past conventional bioassessment. These results demonstrate that targeted hybridization capture enables robust, cross-phylum detection of species used for freshwater bioassessment from environmental DNA.

Background Madagascar has one of the highest burdens of schistosomiasis globally, yet epidemiological evidence from the South-east of the country remains limited. Existing surveillance estimates suggest relatively low prevalence in this region. This contrasts with the region's environmental suitability for transmission and its population's high occupational risk through rice farming. Methods A cross-sectional exploratory study was conducted across 13 villages around the Manombo Special Reserve, Farafangana District, between May and July 2025. Participants were surveyed for geographic, environmental and behavioural risk factors for schistosomiasis infection. Infection was diagnosed by Kato-Katz stool microscopy. Multivariable mixed-effects logistic and linear regression, with village as a random intercept, estimated associations with infection and intensity respectively, adjusting for the water catchment area of household water sources, sex, age group, recent praziquantel treatment and village-level open defecation rate. Results 227 participants were included. Overall S. mansoni prevalence was 53.3% (95% CI 46.8-59.8%); median infection intensity was 96 (IQR 48-288) eggs per gram of stool among those infected. Surface water contact was near-universal (97.3%). Prevalence varied markedly between neighbouring villages. No demographic, behavioural, or WASH variable was independently associated with infection. The water catchment area of household water sources was the primary predictor of infection. Conclusions Prevalence substantially exceeded current surveillance estimates, suggesting this population is underserved by existing control programmes. Water catchment area was the dominant determinant of transmission heterogeneity, possibly consistent with Biomphalaria habitat suitability. Water catchment area may represent a valuable and underutilised unit for targeting schistosomiasis surveillance and control in Madagascar.

Leptospirosis is a globally important environmentally transmitted disease, with approximately one million cases and 60,000 deaths reported annually. In low-income urban communities, inadequate sanitation, drainage and waste management may increase human exposure to contaminated environments. This study investigated the influence of environmental engineering risk factors on Leptospira exposure in four disadvantaged urban communities (favelas) in Salvador, Brazil. A high-precision georeferenced field survey was developed to identify, map and characterise sanitation, stormwater drainage and solid waste infrastructure. Cross-sectional spatial analyses of baseline data were used to assess associations between environmental risk factors and the residential locations of individuals with anti-Leptospira antibodies. Seropositive individuals tended to reside closer to environmental risk factors and at lower relative elevations. Density analyses indicated that seropositive individuals tended to reside closer to inadequate or partially adequate sewerage components than to sewage-contaminated streams or open sewage points. Inadequate streets showed also showed high density peaks, suggesting that exposure may occur through frequent contact with contaminated runoff and standing water. In contrast, open waste dumping sites and vacant lots showed weaker and more diffuse spatial patterns. The findings highlight the importance of infrastructure quality shaping leptospirosis risk within urban informal settlements. The proposed methodology provides a practical approach for high-resolution characterisation of environmental exposure pathways and may support targeted engineering interventions and epidemiological investigations of leptospirosis and other environmentally transmitted diseases.

Wastewater surveillance (WS) offers a non-invasive means of tracking population-level circulation of infectious agents, including viruses linked to cancer. This study provides the first Australian assessment of oncogenic viruses in municipal wastewater by screening 76 influent samples collected over four months from six wastewater treatment plants in Southeast Queensland, Australia. Ten gene targets representing seven oncogenic viruses including Epstein-Barr virus (EBV), hepatitis B virus (HBV), hepatitis C virus (HCV), human herpesvirus 8 (HHV-8), human papillomavirus 16 and 18 (HPV-16 and -18), human T-lymphotropic virus type 1 (HTLV-1), and Merkel cell polyomavirus (MCPyV) were analysed using PCR-based methods. All viruses were detected in wastewater at least once, though with substantial variation in frequency. MCPyV was the most frequently detected virus, appearing in 97.3% of samples with concentrations ranging from 3.09-3.85 log10 gene copies (GC)/50 mL, indicating widespread population exposure. HBV (26.3%) and EBV (15.8%) were detected intermittently across multiple catchments, while HPV-16/18, HHV-8, HTLV-1, and HCV were detected at the lowest frequencies (<8%). This study reports the first baseline dataset for oncogenic viruses in Australian wastewater. More broadly, positive detection of all targeted oncogenic viruses including those associated with low prevalence infections in wastewater demonstrates the potential of WS to complement existing cancer surveillance systems in tracking community-level circulation of these infectious agents.

Micronutrient deficiencies are prevalent in Madagascar, where diets rely heavily on starchy staples and access to animal-source foods is limited. Small dried fish (SDF) are widely available, yet their nutritional value and health risks remain poorly documented. We combined market surveys, taxonomic identification, and micronutrient and heavy metal analyses of nine SDF types collected along National Road 7. The samples encompassed 33 fish families, were dominated by small pelagic species (Clupeidae and Engraulidae), and were appreciated by consumers. A daily portion (5 g for infants; 10 g for young children and women of childbearing age) contributed substantially to Recommended Nutrient Intakes (RNIs). Across samples and groups, SDF were rich (>30% of RNI) in selenium and, for infants and young children, in calcium. All samples were a source of (>15% of RNI), or rich in, phosphorus, whereas iron contributions were more variable but often substantial. Several samples exceeded 100% of RNIs for selenium, calcium, iron, or manganese in infants and young children, and some were also sources of magnesium and, less frequently, zinc. Vitamin A was absent from sun-dried samples but detected in a smoked freshwater type. Heavy metal concentrations varied markedly, and portions of several types led to estimated exposures to inorganic arsenic or cadmium exceeding reference values, whereas freshwater species and some pelagic types showed a more favorable nutrition-risk balance. Overall, SDF are affordable, nutrient-dense foods with strong potential to alleviate micronutrient deficiencies in Madagascar, while highlighting the need for type-specific guidance to balance nutritional benefits and contamination risks.

Context Pumping stations pose a threat to fish globally through land use change, habitat fragmentation and entrainment risk, with the catadromous and critically endangered European eel particularly impacted. Objectives/methods Establish, model, assess and understand the present-day distribution of European eel and resident fishes in 152 pumping station catchments in a once extensive wetland (The Fens) using eDNA metabarcoding (855 samples over two and half years), with specific focus on anthropogenic influences on hydrological connectivity and habitat quality. A removal survey design maximised confidence in negative results while minimising time and consumable costs. Results Eel occurrence upstream of pumping stations was low (occupancy = 28.3%) and positively associated with catchment area, fish species richness and natural hydrological connectivity (gravity drainage or flooding) and negatively associated with distance from the tidal limit. Fish species richness replaced catchment area and improved model performance, potentially acting as a biotic indicator of habitat quality and connectivity. Pumped catchments with manually operated upstream water transfers had reduced eel presence, potentially linked to the direction of water flow or the timing of operation. By contrast, fish species richness increased in these catchments during summer, suggesting displacement into unsuitable long-term habitats. Physical habitat maintenance had no detectable effect on eel occurrence or fish species richness. Conclusions This study provides the first landscape-scale assessment of European eel distribution and drivers of occurrence in pumped river catchments. The highly novel and comprehensive insights have implications for European eel conservation as well as infrastructure and catchment management, including compliance with legislation (EC Regulation No. 1100/2007).

Background In Nigeria, the country with the greatest global malaria burden, funding constraints increasingly require insecticide-treated net (ITN) reprioritization to target those at highest risk. Large language models (LLM)- assisted decision-support tools may facilitate risk-informed ITN planning by supporting malaria programme officers in navigating analyses, interpreting outputs, and translating evidence into operational decisions. We developed ChatMRPT, an LLM-assisted ITN allocation planning tool based on user requirements, and analyzed post-interaction feedback, examined user digital competencies and workplace resources, and identified institutionalization pathways for LLM-assisted intervention planning. Methods A two-phase mixed-methods study began with software requirements gathering workshops (using a prototype) involving representatives from the National Malaria Elimination Programme (NMEP), State Malaria Elimination Programmes (SMEPs), and implementing partners. Phase 2two evaluated ChatMRPT through surveys, guided exercises, and focus group discussions with 34 SMEP officers from 28 Nigerian states. Quantitative data were analyzed using descriptive statistics and profile-based comparisons, while qualitative data were analyzed using reflexive thematic analysis to synthesize user experiences of ChatMRPT and identify institutionalization pathways. Findings Fifty-eight percent (19/33) of participants demonstrated both higher digital competency and adequate workplace resources; the remainder exhibited limitations in one or both domains ([4/33] higher competency/constrained resources; [6/33] higher resources/lower competency). Participants with higher digital competency but constrained workplace resources reported user experiences comparable to those with higher competency and adequate resources, whereas workplace resources alone did not appear to compensate for lower digital competency. Key software requirements included contextual guidance for malaria risk interpretation, operational decision support, and embedded analytical support. Following iterative incorporation of these requirements, ChatMRPT was positively evaluated across participant profiles. Participants viewed institutionalization as dependent on integration into routine malaria planning and adaptability to evolving programme priorities. Interpretation Many malaria programme officers may already have the foundational competency for LLM-assisted decision support. However, there is room to further strengthen digital competencies while facilitating access to basic workplace resources such as stable internet. Institutionalization of LLM tools may depend on addressing these capacity and infrastructural constraints alongside designing explainable, integrated, and flexible systems. Future research should evaluate long-term integration, sustainability, and effectiveness in routine malaria planning. Funding This work was funded by the Bill and Melinda Gates Foundation (INV-036449) and the Center for Health Outcomes and Informatics Research (CHOIR), Loyola University Chicago. The funders had no role in the study design, data analysis, interpretation of findings, or preparation of the manuscript.

Boiling raw milk before drinking is a common practice in Bangladesh, but its impact on residue levels of ciprofloxacin and enrofloxacin is not well known. This study looked at these antibiotics in 120 raw cow milk samples from 12 dairy areas near Dhaka, collected between November 2023 and March 2024. Each sample was split into two parts: one tested as raw milk and the other after boiling at home for 15 minutes. Residues were checked using matrix-matched HPLC-UV. Ciprofloxacin and enrofloxacin were found in 113 out of 120 samples (94.2%), and at least one of them was present in 117 samples (97.5%). We considered a level of 12.5 micrograms per liter or higher as detected. The average combined amount of these antibiotics dropped from 194.61 micrograms per kilogram in raw milk to 179.57 micrograms per kilogram after boiling, a decrease that was statistically significant (p < 0.001). However, the percentage of samples exceeding the EU maximum residue limit of 100 micrograms per kilogram only went down from 92.5% in raw milk to 90.0% after boiling, which was not statistically significant (p = 0.250). Using national milk consumption data as an estimate of intake, the hazard index for adults was 0.360 for raw milk and 0.332 for boiled. Calculated with a 10 kg body weight, these values were 2.159 and 1.992, respectively. Overall, boiling at home did reduce the levels of ciprofloxacin and enrofloxacin, but it usually did not bring high-residue samples below the EU limit.

Many Sockeye salmon (Oncorhynchus nerka) populations have declined over recent decades, and climate change is likely to exacerbate these declines through direct and indirect ecological effects. The response to the associated environmental changes is likely to vary among life stages, populations, and regions. Quantitative estimates of climate change driven impacts that account for this variability could fill a critical gap and provide forward-looking insights into how sockeye are expected to respond to future climate-driven change across their lifecycle. To address this need we developed a hierarchical population dynamics model parameterized with juvenile, adult return and spawner abundance data from 13 sockeye salmon populations from Washington State to northern British Columbia. We used a formal causal inference framework that paired salmon abundance data with a suite of environmental covariates hypothesized to represent ecological conditions across the lifecycle. We used the model to estimate population-specific responses to each environmental driver, then combined parameter estimates with projections from down-scaled climate change models to estimate productivity responses to anticipated environmental change. We found that historical sockeye productivity was strongly associated with environmental covariates, which explained more interannual variability in return abundance than spawner abundance in most populations. However, the life stages and specific environmental covariates with the largest impacts differed among populations and regions, often displaying a latitudinal gradient. Increases in coastal ocean temperatures and mixed layer depth generally had negative effects though they varied among regions. Increased freshwater summer rearing and return migration temperatures had weaker but consistently negative effects. Under future climate conditions, projected changes in these environmental covariates are expected to result in substantial declines in productivity across most populations. Sockeye salmon display varying degrees of sensitivity to climate change across life stages, populations, and regions. Effective future management will require explicitly accounting for these life stage and population-specific responses.

Rinsing-dependent dental hygiene presents a significant public health challenge in water-scarce environments. This study investigated combinations of xylitol (Xyl), chitosan (Chi), glycyrrhizin (Gly), epigallocatechin gallate (EGCG), dicalcium phosphate (DCP), and nano-hydroxyapatite (nHA) on the primary bacteria behind dental caries, S. mutans. These combinations were assessed for markers of dental caries by biofilm reduction, bacterial killing, and acid buffering against S. mutans when applied to an in vitro simulated enamel model using glass bead surfaces for biofilm formation, and gene expression was subsequently examined via RT-qPCR. Separately, mineral retention was also quantified. The EGCG-DCP-Xyl film demonstrated the highest overall efficacy, achieving a significant reduction in biofilm concentration compared to the untreated control and performing similarly in magnitude to the positive toothpaste control. Dead fluorescence staining confirmed that the EGCG-DCP-Xyl film induced the highest rate of non-viable cells, followed by the Chi-Gly film and the Gly-Xyl film. During 10-day pH cycling, the EGCG-DCP-Xyl and DCP-Xyl formulations buffered pH the most, consistently maintaining mean pH levels safely above the demineralization threshold of pH 5.5. The EGCG-DCP-Xyl also optimized mineral stability with the highest retained calcium concentration, significantly outperforming the Chi-Xyl film. At the transcript level, the EGCG-DCP-Xyl film induced substantial downregulation of key virulence genes, yielding decreases in expression for glucosyltransferase B (gtfB), associated with biofilm synthesis, collagen-binding protein (cnm), associated with tissue invasion, and lactate dehydrogenase (ldh), associated with lactic acid production, compared to the untreated control, with effects comparable in magnitude to the positive toothpaste control. This research suggests that targeting bacterial pathways and mineral loss through a portable film may have potential for preventing dental caries, especially in environments where water is limited. However, additional studies are necessary to evaluate real-world effectiveness.

Background: In two large studies conducted in Bangladesh, our recently developed artificial intelligence (AI)-based models for assessing dehydration severity in children under five years (DHAKA models) and patients over age five (NIRUDAK models) were significantly more accurate and reliable than the WHO IMCI and IMAI guidelines for diarrhea management. We incorporated these models into a novel mobile health (mHealth) clinical decision support tool (CDST), called FluidCalc, with the potential to improve acute diarrhea management by frontline health workers worldwide. Our objective was to assess the barriers and facilitators to uptake and use of our mHealth CDST in both a low-resource setting (Tanzania) and high-resource setting (United States (US)) among healthcare providers and stakeholders. Methods: Qualitative data were collected through focus group discussions (FGDs) with healthcare providers and in-depth interviews (IDIs) with stakeholders and policymakers from February - July 2025 in Tanzania and February - March 2026 in the US. The Consolidated Framework for Implementation Research (CFIR) was used to guide discussions and elicit participant feedback. Audio recordings were transcribed and translated from Swahili to English where applicable, and data were analyzed using framework matrix analysis. Results: 35 providers from different cadres participated in FGDs, and 13 stakeholders participated in IDIs. Facilitators to implementation included FluidCalc's simplicity, ease of use, and offline functionality. Participants reported that the app could streamline clinical workflows, promote adherence to diarrhea management guidelines, facilitate task shifting, support antibiotic stewardship, and reduce errors in fluid rehydration calculations. FluidCalc was also viewed as a valuable teaching tool, and for supporting less experienced healthcare providers and trainees, and as useful during diarrheal disease outbreaks. Perceived barriers included the need for reliable digital infrastructure, including access to mobile devices, internet connectivity, and dependable electricity and lengthy institutional approval processes. Endorsement and approval from the Ministry of Health and health facility leadership were perceived as essential for successful implementation. Conclusion: Healthcare providers and stakeholders believe FluidCalc has the potential to improve care for patients with acute diarrhea in both high- and low-resource settings. Addressing identified barriers and ensuring reliable digital health infrastructure are needed to support effective integration into patient care.

Background Scientific mistrust contributes to lower participation and underrepresentation of Black Americans in genomics studies limiting understanding of how genomic variation and environmental exposures influence health disparities. Community-engaged research requires rebuilding scientific trust; however, there is a need for practical models that operationalize guidance for researchers without community-engaged research training. Therefore, we developed an educational intervention for genomics researchers initiating partnerships with Black American communities. Our intervention creates a bidirectional teaching environment that allows potential community and academic partners to discuss areas of expertise for each stakeholder, partnership perspectives and needs, while exploring modules related to genomics research and community-academic partnership. We report a novel and structured approach for prospective academic and community partners to mutually orient one another and assess partnership practicability. Methods Ten participants, recruited through established community channels, attended a four-hour workshop containing nine interactive modules about scientific mistrust, research safeguards, community-engaged research, genomics, and research for community-defined goals. We also experimented with humor to enhance engagement and trust. We used mixed-methods, single-arm research design and assessed feasibility through recruitment success and retention. Using inductive rapid thematic analyses, we assessed participant responses to eight workshop prompts. Preliminary quantitative data, used for descriptive purposes due to low sample size, were analyzed from pre- and post-Likert scale surveys assessing associations between the intervention and four domains, including scientific trust. Results All 10 enrolled participants completed the study, meeting a priori criteria of 100% for recruitment and show rate. Engagement was strongest during early workshop modules and declined in later modules. Survey data completeness was limited by missing responses. Survey instrument design limitations were identified for modification in future studies. Participants articulated expectations for partnership that aligned with community-based participatory research principles. Conclusions The intervention is feasible to deliver as a bidirectional educational experience in partnership with a community organization. The strongest implementation refinements needed were workshop duration, module prioritization, and survey instrument design, particularly the trust domain. A community advisory board is co-developing modules and refining the intervention to evaluate in a pilot version of the study with a larger sample.