Environmental Science & Technology

Traditionally, studies have explored the impacts of individual water chemistry parameters on the persistence of Mycobacterium spp. and Legionella spp. in isolation with the underlying assumption that these associations are likely monotonic in nature. Yet chemical and microbiological changes are complex, and associations are likely highly combinatorial. In this study, we use interpretable machine learning models to disentangle the integrative and nonlinear associations between water chemistry and occurrence/abundance of Mycobacterium spp. and Legionella spp. Seasonal data from source water, point-of-entry and distribution systems of eight full-scale drinking water systems demonstrated that shifts in overall water chemistry were associated with the changes in microbial abundance during treatment and distribution. Machine learning models indicated moderate predictive ability of integrated water chemistry towards Legionella spp. abundance and towards the occurrence of both Legionella spp. and Mycobacterium spp., whereas predictive performance for Mycobacterium spp. abundance was limited. The association between nitrate and Legionella spp. abundance was disinfectant regimes dependent, while dissolved organic carbon exhibited a concentration dependent response type (i.e., positive and negative association). In chloraminated systems, Legionella spp. abundance was positively associated with ammonia and nitrate, highlighting the critical role of nitrification. Here, it appears that pH likely influences the initial colonization of Legionella spp. while ammonia governs its abundance in drinking water. Overall, this study demonstrates that integrated water chemistry and parameter-specific nonlinear effects collectively explain persistence of Mycobacterium spp. and Legionella spp. in drinking water systems.

Neem-derived biopesticides are increasingly applied in agriculture and have been tested in aquaculture research, yet their effects on non-target aquatic invertebrates remain insufficiently characterized. We evaluated the effect of neem extract on the brine shrimp Artemia franciscana using an integrated ecotoxicological approach combining phenotypic, transcriptomic, and histological analyses. Juvenile A. franciscana exhibited dose-dependent mortality and sublethal abnormalities, with a 24 h median lethal concentration of 292.48 mg/L (95% confidence interval, 257.75- 331.89) for mortality and a median effective concentration of 146.36 mg/L (95% confidence interval, 113.04- 189.50) for the combined endpoint "abnormal + dead". In adults, males showed greater mortality than females after extended exposure. High-throughput RNA sequencing revealed broad treatment-associated differences in transcript abundance, with juveniles displaying downregulation of detoxification enzymes and chitin biosynthesis genes, alongside enrichment of immune- and cuticle-related gene ontologies. Adults showed transcriptional signatures of stress, including upregulation of heat shock proteins and cytoskeletal components, and suppression of genes involved in energy metabolism. Chitin precursor enzymes were selectively downregulated in males, and altered carbohydrate metabolism was observed in females. Histological analyses revealed structural deterioration of the brood sac cuticle and reduced ovarian area in treated females, consistent with transcriptomic evidence of impaired exoskeletal and reproductive processes. Overall, neem exposure was associated with phenotypic, histological, and transcriptomic changes in A. franciscana. These results support the use of combined transcriptomic and histopathological endpoints to characterize responses to plant-derived biopesticides in aquatic arthropods.

Formaldehyde is a highly toxic metabolite that can cause extensive damage to DNA and proteins, and strategies to mitigate formaldehyde toxicity are poorly understood. Methylotrophic bacteria, such as Methylobacterium extorquens, thrive on one-carbon compounds as sole sources of carbon and energy. These organisms are excellent models for discovering formaldehyde stress response systems because formaldehyde is an obligate intermediate in their central carbon metabolism. Here, we characterize an evolved def allele (defevo) that increases formaldehyde resistance in M. extorquens. The def gene encodes peptide deformylase (PDF, EC:3.5.1.88), an enzyme that contributes to protein processing by removing the formyl group from N-formylmethionine (fMet) on nascent peptides. The defevoallele has a single missense mutation that decreases PDF activity both in vitro and in vivo. Transcriptomic analysis of the defevo strain indicates there are pleiotropic effects of this mutation and a differential response to formaldehyde stress. We investigate possible mechanisms for the defevo mutants increased resistance to formaldehyde, including mitigation of formaldehyde-induced protein stress and altered membrane physiology. We find that the defevo allele selectively alleviates exogenous, but not endogenous, formaldehyde stress and identify a tradeoff in heat shock resistance. This study reports the first observation of lowered PDF activity benefiting a cellular physiological phenotype. Our work indicates that altered protein metabolism can mitigate the toxic effects of formaldehyde and furthers our understanding of the strategies that can protect cells from formaldehyde-induced damage. ImportanceFormaldehyde is a toxic chemical that can damage essential molecules inside of cells, yet all organisms inevitably produce it during normal metabolism. Despite its ubiquity, our understanding of strategies for how cells navigate formaldehyde toxicity is incomplete. This study focuses on Methylobacterium extorquens, which naturally generates high levels of formaldehyde as part of its growth on simple carbon compounds. We show herein that a single genetic change, which slows down how newly made proteins are processed during translation, can unexpectedly improve the bacteriums ability to resist formaldehyde stress. Further, we show that this single change has numerous effects on the cell, many of which may contribute to formaldehyde resistance.

Indo-Pacific humpback dolphin (Sousa chinensis) and finless porpoise (Neophocaena phocaenoides) are increasingly threatened across their native range, yet the relative influence of multiple stressors in shaping their population dynamics remains unclear. Current conservation strategies for both species are limited by incomplete data and limited assessment of affecting factors. Here, we integrated eDNA metabarcoding with Joint Species Distribution Modeling (JSDM) to assess how environmental gradients, pollution, and trophic associations interactively influence cetacean distributions in Hong Kong waters. We show that degraded water quality and intensified human activity negatively associated with cetacean occurrence, with clear species-specific responses to different stressors. S. chinensis covaried most strongly with Secchi disc depth, and presence of vessels, while N. phocaenoides was negatively associated with nitrate nitrogen and microbial pollution (sewage). The JSDM variance partitioning analysis highlighted that the occurrence of S. chinensis was primarily associated with anthropogenic disturbances (30.04%), followed by water physical properties (26.63%), whereas N. phocaenoides was more strongly associated with physical (40.9%) and anthropogenic disturbances (35.2%). By integrating eDNA and JSDM, our approach provides fine-scale diagnostics of species-specific vulnerabilities, supporting adaptive conservation strategies and guiding the realignment of protected areas to mitigate biodiversity loss in urbanized marine ecosystems. Environmental ImplicationOur study demonstrates that hazardous water pollutants, including microbial contamination, nutrient enrichment, and chemical stressors, vessel pressure, show strong, species-specific impacts on resident cetaceans in Hong Kong. By integrating eDNA metabarcoding with joint species distribution models, we provide a diagnostic framework that directly links pollutant profiles to ecological risk. These findings highlight that conventional conservation strategies overlooking pollution drivers are insufficient for marine megafauna persistence. Our approach offers an early-warning system for monitoring hazardous pollutants in coastal ecosystems and supports adaptive management strategies to mitigate biodiversity loss in urbanized seascapes.

Phenazines are redox-active microbial metabolites produced and secreted in diverse ecological contexts from soils to chronic infections. In these disparate environments phenazines can function variously as antibiotics, extracellular electron shuttles, and nutrient scavengers. Key to understanding the impact of these functions is a robust expectation of phenazine retention or diffusion in a given context. But predicting phenazine fate and transport is difficult because of the chemical complexity of their local microenvironments. To address this challenge, we measured the octanol water distribution coefficient (LogD) as a proxy for lipophilicity of three naturally occurring phenazines produced by the opportunistic pathogen Pseudomonas aeruginosa: phenazine-1-carboxylic acid, phenazine-1-carboxamide, and pyocyanin. We investigated the behavior of both oxidized and reduced forms of these phenazines across broad ionic strength and pH conditions. While the ionic context exerts only small effects, the pH and redox state contribute strongly and independently to changes in phenazine lipophilicity. The pH trends are expected, but the observed redox dependence is generally missed by existing lipophilicity calculation methods. Additional LogD measurements with 1-hydroxyphenazine and unsubstituted phenazine, together with density functional theory modeling of phenazines in their reduced and oxidized forms, reveal that intramolecular hydrogen bonding contributes significantly to the increased lipophilicity of reduced phenazines that possess H-bond accepting substituents in the 1-position. These results explain phenazine behavior in a biological context: redox state alone significantly alters retention of pyocyanin in planktonic P. aeruginosa cells, with the reduced species being predominantly retained by membranes. We propose that the modulation of phenazine lipophilicity in response to the local redox environment has evolved to give a competitive advantage to bacteria by retaining or dispersing these bioactive molecules. Beyond improving our understanding of natural phenazine fate in diverse microbial contexts, our results emphasize an oft-overlooked theme relevant to rational drug and electrochemical shuttle design: redox state matters.

Background Polycyclic aromatic hydrocarbons (PAHs) and volatile organic compounds (VOCs) are combustion-derived pollutants linked to cardiovascular disease. Prior NHANES analyses have evaluated these chemicals individually, failing to capture the correlated co-exposure structures that characterize real-world environmental burden, thereby underscoring the need for application. In this study, we applied an unsupervised machine learning pipeline to urinary biomarker data to identify multi-chemical exposure clusters and quantify their differential cardiovascular risk profiles in a nationally representative US sample. Methods We analyzed 2,979 participants from NHANES between 2017-2018, representing an estimated 36.8 million US adults after complex survey weighting. Twenty-five urinary biomarkers (6 PAH, 19 VOC metabolites) were log-transformed, imputed using Multivariate Imputation by Chained Equations (MICE), and standardized. Uniform Manifold Approximation and Projection (UMAP) was used for dimensionality reduction, followed by Gaussian Mixture Model (GMM) clustering. Survey-weighted prevalence estimates with 95% confidence intervals (CIs) were calculated for hypertension and high total cholesterol within each cluster. Weighted multivariable logistic regression was used to estimate odds ratios (OR) for hypertension, adjusting for age, sex, race/ethnicity, and income. Results Four exposure clusters were identified with a mean assignment probability of 0.948. The High combustion cluster (n=370; estimated 5.1 million US adults) exhibited the highest multi-chemical burden and a weighted hypertension prevalence of 39.3% (95% CI 37.2-41.4%), compared to 28.7% (95% CI 21.9-35.5%) in the Low exposure reference group. After demographic adjustment, High combustion cluster membership was independently associated with 38.4% higher odds of prevalent hypertension (OR 1.38). The prediction model achieved a cross-validated area under the receiver operating characteristic curve (AUC) of 0.849 (SD 0.017). Non-Hispanic Black participants constituted approximately 40% of the High combustion cluster, exceeding their representation in lower-risk clusters. Conclusions Multi-chemical exposome profiling identifies four cardiovascularly distinct subpopulations in the US adult population. Membership in the High combustion exposure cluster was associated with higher odds of prevalent hypertension and disproportionately affected Non-Hispanic Black participants. These findings support the use of multichemical approaches over single-pollutant analyses and highlight the relevance of environmental exposure patterns for making policy and targeted cardiovascular risk stratification.

Metataxonomic analysis targeting the V4 region of the 18S rDNA gene, combined with molecular phylogenetic inference, was applied to detect nematode DNA of public health relevance in environmental matrices. A total of 25 mOTUs corresponding to six nematode taxa were detected in environmental samples from the Andean region of Colombia. Analysis of 12 water and sludge samples from wastewater treatment plants, 5 artisanal agricultural bioinputs, and 3 food samples revealed multiple species of public health significance: Trichuris trichiura, Enterobius vermicularis, Ascaris spp., and Necator americanus. We also confirmed zoonotic species, including Angiostrongylus cantonensis and Trichinella spp. These findings demonstrate that combining metataxonomics with molecular phylogeny provides a scalable molecular framework for the environmental surveillance of parasitic nematodes, overcoming the limitations of traditional morphological identification methods. This approach offers a replicable model for strengthening control and monitoring programs for parasitism in human populations.

Wastewater surveillance is increasingly used for antimicrobial resistance (AMR) monitoring in urban environments, but low-resource settings often lack a piped sewerage system. Instead, coprophagous flies--flies that ingest feces--may serve as composite samplers for monitoring fecal wastes present in terrestrial environments. We evaluated whether the class 1 integron-integrase gene intI1 was associated with genetic markers of AMR and fecal source tracking markers (FST) in coprophagous flies collected from latrine entrances and food preparation areas in low-income urban Maputo, Mozambique. We quantified intI1, an enteric 16S rRNA target (for normalization), three FST markers, and 30 ARG targets using qPCR. We normalized concentrations of intI1 and each target to enteric 16S rRNA. We fit linear mixed models with a random intercept for housing compound to estimate within-fly associations between log10 relative abundance of intI1 and log10 relative abundance of each target with and without adjustment for fly taxonomic group, capture location, and standardized fly mass. We also modeled per-fly unique ARG count (i.e., number of ARG targets detected) using Poisson regression. Of 188 flies assayed, 176 passed internal controls; intI1 and enteric 16S rRNA were detected in 95% and 96% of flies, respectively. Higher relative abundance of intI1 was positively associated with ARG and FST targets, with the strongest associations observed for sulfonamide-(sul1: {beta} = 0.87; 95% CI: 0.81, 0.94; sul2: {beta} = 0.81; 95% CI: 0.73, 0.89), tetracycline- (tetA: {beta} = 0.78; 95% CI: 0.70, 0.85; tetB: {beta} = 0.69; 95% CI: 0.60, 0.79), and trimethoprim-related (dfrA17: {beta} = 0.78; 95% CI: 0.70, 0.86) genes. Associations with FST markers were weaker (i.e., human mtDNA: {beta} = 0.46; 95% CI: 0.37, 0.55; human-associated Bacteroides: {beta} = 0.34; 95% CI: 0.25, 0.43). Higher relative abundance of intI1 was also associated with a greater number of ARGs detected: each 10-fold increase in intI1 was associated with an 8% higher expected unique ARG count (aRR=1.08, 95% CI: 1.04-1.12). These findings support the need for further research across different settings exploring intI1 carried by coprophagous flies as a potential standardized screening target for AMR surveillance in unsewered terrestrial environments.

Browning and eutrophication strongly influence aquatic ecosystems by altering nutrient dynamics, light availability, and food web structure. To investigate their combined effects on aquatic communities, we conducted a nine-week mesocosm experiment in a clear-water north-temperate lake, crossing dissolved organic carbon (DOC) and total phosphorus plus total nitrogen (TP+TN) enrichment treatments. Multi-trophic plankton communities (bacterioplankton, phytoplankton, and zooplankton) were monitored over time using environmental DNA (eDNA) marker gene amplicon sequencing. Beta-diversity analyses highlighted temporal and treatment-driven community restructuring, while PERMANOVA and Principal Response Curve analyses identified the treatments and taxa driving these changes. Our results show that elevated DOC favoured taxa associated with the microbial loop, while nutrient enrichment and lower DOC promoted the green pathway. Threshold responses across trophic levels were observed at 5-7 mg L-{superscript 1} DOC and 30-70 g L-{superscript 1} TP, marking the levels at which compositional shifts propagated through the food web. Overall, this study demonstrated how aquatic communities respond dynamically to browning and nutrient enrichment, offering insight into the mechanisms shaping multi-trophic interactions under a multiple stressor scenario. HighlightsO_LIBrowning and nutrient pulses drove coordinated shifts across bacterioplankton, phytoplankton, and zooplankton. C_LIO_LITemporal community succession and treatment effects were captured through beta-diversity and multivariate ordination analyses. C_LIO_LIThreshold responses propagated through the food web at 5-7 mg L-{superscript 1} DOC and 30-70 {micro}g L-{superscript 1} TP. C_LI Scientific Significance Statement TopicThis study provides insights into the interactive effects of browning and eutrophication on community composition shifts in freshwater ecosystems. Using a mesocosm experiment, we identified thresholds for dissolved organic carbon and total phosphorus + nitrogen concentrations that drove compositional changes across bacterial, phytoplankton, and zooplankton communities, as determined by environmental DNA amplicon sequencing data. Graphical Abstract O_FIG O_LINKSMALLFIG WIDTH=200 HEIGHT=133 SRC="FIGDIR/small/719940v1_ufig1.gif" ALT="Figure 1"> View larger version (22K): org.highwire.dtl.DTLVardef@5f11dborg.highwire.dtl.DTLVardef@18d4f6aorg.highwire.dtl.DTLVardef@58cb68org.highwire.dtl.DTLVardef@1924a42_HPS_FORMAT_FIGEXP M_FIG C_FIG

Biodiversity is expected to enhance the stability of ecological communities under environmental stress, but the relative roles of functional, interspecific, and intraspecific diversity remain poorly resolved, particularly under multiple concurrent stressors. We tested how these diversity dimensions shaped the growth and recovery of marine Synechococcus communities in a microcosm experiment manipulating strain composition across four strain-richness levels and two interspecific diversity levels under control, atrazine, warming, and combined atrazine-plus-warming treatments. Functional diversity was quantified from flow-cytometric trait data and analyzed as initial functional diversity during the stress phase and assembled functional diversity during recovery. Contrary to our expectations, higher initial functional diversity was associated with lower community growth during stress, while higher assembled functional diversity was generally associated with weaker recovery. However, these relationships depended on stressor identity and interspecific diversity: in two-species communities, the negative effects of functional diversity were reduced, and under combined stress, higher assembled functional diversity was associated with improved recovery. In contrast, intraspecific diversity consistently enhanced community growth and recovery, while interspecific diversity primarily promoted functional recovery. Together, our results show that functional, interspecific, and intraspecific diversity can influence stress responses through distinct pathways.

Ocean warming is currently leading to distributional shifts of species and an alternation of coastal communities. Vulnerable species that are most sensitive to ocean warming are able to use several acclimation mechanisms, with one of the fastest being a shift in and shuffling of their partnerships with symbiotic microorganisms. Assessing symbiosis-focused mechanisms of acclimation and adaptation in response to ocean warming is a technical challenge due to the difficulty of accurately simulating the de novo formation of coastal communities. Here, we use the Kiel Outdoor Benthocosm facility to assess which sponges species are experimentally recruited and whether they exhibit symbiosis-focused mechanisms of acclimation following selection to ocean warming. We observed one sponge species (Haliclona sp.) and found that this sponge exhibited significant shifts in the membership and composition of its associated microbiome in response to ocean warming, with much of this being attributed to the rare microbiota. Moreover, Haliclona sp. maintained the diversity and dominance of its microbiome members. Four bacteria taxa were differentially abundant at elevated temperatures, with two being a Francisella sp. that is a suspected pathogen and an uncultured Francisellaceae that is most closely related to sulfur-oxidizing endosymbionts. Changes to the Haliclona sp. microbiome are largely consistent with a limited acclimation response, which could indicate that this sponge may use microbial symbionts as part of a mechanism to acclimate and adapt to a warmer future ocean.

IntroductionDrowning risk begins with water exposure, yet population-water relationships have rarely been quantified at scale using environmental measures. This study explored whether satellite-derived data was associated with subnational drowning mortality and whether associations differed by country income level. MethodsWe linked Global Burden of Disease (GBD 2021) age-standardised drowning mortality rates to satellite-derived exposures for 212 subnational regions across 12 countries (2006-2021; 3,392 region-years). Exposures were extracted via Google Earth Engine and standardised. Gamma-log generalised linear mixed models included region random intercepts and year fixed effects. Income-stratified models were estimated separately. Supplementary models assessed maritime vessel activity. ResultsNear-water population percentage was the strongest correlate of drowning (IRR 1.40; 95% CI 1.33-1.47). Permanent water coverage was protective (IRR 0.80; 0.73-0.88), as were nighttime lights (IRR 0.96; 0.95-0.97) and hot days [≥]30{degrees}C (IRR 0.95; 0.92-0.99). Mean temperature (IRR 1.17; 1.11-1.23) and precipitation (IRR 1.03; 1.01-1.04) were positively associated. Near-water effects were consistent across income strata (LIC 1.25; MIC 1.31; HIC 1.24), while other predictors showed weak or inconsistent within-strata associations. Vessel activity was modestly associated with drowning in Global Fishing Watch models (IRR 1.05; 1.01-1.09) but not in Synthetic Aperture Radar models. DiscussionSatellite-derived indicators can characterise drowning risk at scale, with population proximity to water emerging as a robust cross-context correlate. Protective associations for permanent water suggest landscape configuration may shape risk beyond proximity alone, highlighting geospatial datas value for targeting prevention where surveillance is limited.

Drowning remains a major global public health challenge, yet how built environment characteristics shape population-level drowning risk remains poorly understood. This study linked satellite-derived built environment data to subnational drowning mortality estimates across 203 regions in 12 countries from 2006-2021. It found that built environment associations with drowning mortality are complex, non-linear, and shaped by development context. Urban extent was strongly protective, while built area near water showed protection overall but increased risk when combined with high population crowding. Almost all drowning mortality variance occurred between regions rather than within regions over time, indicating risk is predominantly determined by place-based characteristics. Income-stratified analyses revealed profound heterogeneity: crowding was protective in low-to middle-income settings but near-null in high-income regions, while waterfront development captured very different realities across contexts. These findings highlight the importance of tailoring drowning prevention strategies to local built environment configurations and development contexts.

The increasing use of microbial biopesticides in sustainable agriculture requires a deeper understanding of their potential impact on non-target pollinators. Although biocontrol agents are generally considered safer than synthetic pesticides, they may still cause subtle but ecologically relevant adverse effects on non-target organisms, especially when exposed to multiple stressors that are often overlooked in current risk assessment frameworks. Among these, nutritional stress, caused by habitat loss, fragmentation and reduced floral diversity, is becoming increasingly widespread. In this study, we investigated the lethal and sublethal effects of the bacterial biopesticide Bacillus velezensis (formerly B. amyloliquefaciens) strain QST713 at field-relevant concentrations on two key pollinators: Apis mellifera and Bombus terrestris. For the first time for a biopesticide, oral toxicity was assessed under environmental stress represented by diets with varying sugar concentrations (optimal and suboptimal) to identify potential synergistic effects on bee health. Sublethal effects were examined by studying learning performance and memory retention through a conditioning experiment under laboratory conditions. The results showed marked species-specific differences. While B. velezensis did not impact bee survival under realistic nutritional conditions, we observed a synergistic lethal effect in B. terrestris when biopesticide exposure was coupled with extreme nutritional stress (sugar deprivation). Similar species-specific differences emerged at the behavioral level: unlike A. mellifera, B. terrestris showed impaired visual learning and early long-term memory recall. Taken together, these results show that sublethal cognitive endpoints and multi-stressor contexts may reveal vulnerabilities not immediately evident through mortality-based assessments alone. Our findings also highlight the importance of including multiple pollinator species in risk assessment, as sensitivity to biopesticides might greatly vary among species.

Jeevamrit, a fermented liquid microbial bioinoculant, is increasingly recognized as a soil and plant growth enhancer in sustainable agricultural practices such as zero-budget natural farming; however, the genetic pool attributed to the functionality by microbial constituents remained poorly resolved. In this study, we reconstructed 16 high-quality metagenome-assembled genomes (MAGs) from Jeevamrit under two critical mixing regimes to elucidate the contributions of key taxa affiliated with Pseudomonadota, Bacillota, and Bacteroidota to nutrient cycling and plant growth promotion. Functional annotation revealed a stratified (upper-middle-lower) metabolic organization with interdependent interactions driving combinatorial functionality. Upper-layer MAGs, including Klebsiella and Pseudaeromonas exhibited organic polymer degradation, glycolytic and oxidative carbon metabolism, respiratory versatility with nutrient acquisition traits such as nitrogen fixation and phosphate/iron solubilization. In middle and lower-layer, Trichococcus, Clostridium, and Veillonella displayed fermentative and reductive metabolisms that facilitate the turnover of partially degraded organic matter and production of organic acids, nitrogen transformations, and metabolic cross-feeding under fluctuating redox conditions. Phylogenetic and taxono-genomic analyses support the designation of eight MAGs as novel species (sp. nov.), for which new names are proposed. A consensus genetic map deciphered traits linked to phytohormone biosynthesis (IAA, cytokinins), quorum-sensing-mediated rhizosphere colonization, and abiotic stress tolerance. Ultimately, this culture-independent metagenome study underpins field-relevant mechanistic insights into an indigenous microbial inoculant, highlighting its potential as a locally adapted solution for sustainable agriculture. ImportanceMicrobial bioinoculants such as Jeevamrit are increasingly used in sustainable agriculture, yet their functional basis remains insufficiently understood due to the limited genome-level resolution of constituent microbiota. This study addresses this gap by applying genome-resolved metagenomics to connect microbial diversity with agriculture-associated ecological functions in a complex fermented local formulation. By integrating metabolic reconstruction with plant-associated functional traits, this study advances understanding of how microbial consortia contribute to nutrient mobilization, rhizosphere competence, and environmental adaptability. This highlights the contribution of yet-to-be-cultivated but metabolically versatile taxa, which are important to the functions of agricultural ecosystems. By uncovering the roles of key microbes and their cooperative metabolic interactions, this work provides a scientific basis for improving Jeevamrit formulations through informed selection or enrichment of functionally important microbes to enhance nutrient delivery and plant growth performance.

Returning individualized microbiome results in ways that are ethical, comprehensible, and useful remains under-explored in African settings. We nested a multi-site, mixed-methods study within the AWI-Gen Wave 2 gut microbiome sub-study of 1,801 women aged 42 to 86 years to engage the participants and provide feedback. All (1,001) participants from Agincourt and Soweto (South Africa) and Nairobi (Kenya) were invited to feedback meetings: 496 from Agincourt, 87 from Soweto, and 195 from Nairobi responded. Engagement strategies were tailored by site (small-group and home-based sessions, visual metaphors, Foldscopes, and local-language delivery). Using semi-structured discussions and structured observations analysed thematically in MAXQDA under COREQ, five cross-cutting themes emerged: (1) understanding of microbiome reports, (2) emotional responses to feedback, (3) perceived health relevance, (4) trust in research institutions, and (5) suggestions for improving engagement. Culturally grounded explanations and local-language facilitation enhanced comprehension and perceived relevance; English-heavy sessions were associated with more confusion. Most participants expressed satisfaction and described planned or enacted dietary and lifestyle changes, while frustration centred on long delays between sampling and feedback. Trust increased with transparency and individualized return of results but was often conditional on minimizing burdensome procedures such as repeat blood sampling (phlebotomy) and ensuring timely feedback. Engagement was feasible and low-cost (approximately USD 29-59 per participant) with site-specific resource needs. Limitations included constrained generalizability beyond the three study sites. Returning individualized microbiome findings in community settings in Africa is acceptable, feasible, and can motivate health-promoting behaviours when delivered promptly and in culturally and linguistically appropriate ways.

Respiratory virus infections affect both humans and livestock, causing considerable mortality and morbidity. While respiratory pathogens such as swine influenza A virus (pH1N1) and porcine respiratory coronavirus (PRCV) often present with overlapping clinical symptoms, their pathological trajectories and outcomes differ. Given the propensity for pathogen spillover and the use of pigs as a physiologically relevant large-animal translational model, we aimed to characterise host serum protein signatures that detect and differentiate pH1N1 from PRCV, enabling improved disease monitoring and control. Using high-resolution mass spectrometry- based proteomics, we identified 162 serum proteins that were significantly dysregulated across 3 infection timepoints (1, 5, and 12 days post-infection (DPI)), with signatures correlating with viral shedding and lung pathology as early as 1 DPI. Notably, multiplexed targeted analysis of a subset of proteins in an independent cohort from a different breed and geographic location demonstrated detection, femtomole-level targeted quantitation, and validation of SRGN as a diagnostic marker for pH1N1 and PRCV (AUC=0.85). Further, SOD1 was validated as an early marker for PRCV, increasing as early as 1 DPI (AUC= 0.9). Finally, a multi-peptide signature composed of SRGN, SOD1, and RAN demonstrated reasonable predictive power for pH1N1 (AUC=0.75) and PRCV (AUC=0.65) at 1 DPI. Our data validate the proteomic screening, provide insights into the role of early protein markers in distinguishing respiratory viral infections, and pave the way for the development of point-of-care diagnostics and targeted prevention strategies, enhancing preparedness against emerging zoonotic threats.

Nowadays N95 facial mask has gain huge attention due to COVID19 pandemic situation and it serves as the prime PPE. Though the microbes can be restricted to get inside the human body due to the presence of mask temporarily, but over the time, bacteria and other microbes may get entrapped into the threads of the mask itself and thus acting as a storage chamber of microbes. It is necessary to eliminate them from the mask surface. To do so different floral structured Nano-ZnO with variable oriented arrangement of petals were fabricated on the surface of the N95 mask and further characterized through instrumentations including XRD, FTIR,UV-Vis, Fluorescence-Spectroscopy, SEM, DLS. The average crystallite size calculated for synthesized four different ZnO nanoflower were 25.19 nm, 23.46 nm, 27.27 nm and 31.78 nm (for glycerol, PEG, EDTA, Chitosan assisted) respectively. The antimicrobial activity was investigated by standard microbial broth dilution assay and Kirby-Bauer test which assured the inhibition of the bacterial growth. The MIC-MBC value of ZnO nanoflowers for E.coli and B. subtilis were found to be effective at dilution of 250 {micro}g/ml and 100 {micro}g/ml. Additionally a modified Kirby-Bauer assay has been designed to investigate the killing efficiency of the bacteria (E.coli). O_FIG O_LINKSMALLFIG WIDTH=200 HEIGHT=145 SRC="FIGDIR/small/719592v1_ufig1.gif" ALT="Figure 1"> View larger version (35K): org.highwire.dtl.DTLVardef@48e5ecorg.highwire.dtl.DTLVardef@1ef03c5org.highwire.dtl.DTLVardef@e089ddorg.highwire.dtl.DTLVardef@17b2850_HPS_FORMAT_FIGEXP M_FIG O_FLOATNOFig. - Graphical AbstractC_FLOATNO C_FIG

Extracellular vesicles (EVs) are lipid bilayer-enclosed particles that mediate intercellular communication through the transfer of bioactive molecules. Their growing relevance in translational applications demands downstream purification workflows that are selective, scalable, and compatible with robust impurity control. Conventional EV isolation methods primarily rely on physicochemical properties such as size, density, or charge and therefore co-enrich overlapping EV fractions together with non-vesicular impurities. Here, we establish a Nanofitin(R)-based affinity chromatography workflow for selective enrichment of a CD81-positive EV fraction under EV-compatible elution conditions. Nanofitin(R) candidate NF06 was identified by ribosome display against the large extracellular loop of CD81 and combined nanomolar affinity with favorable release behavior while retaining binding after repeated regeneration cycles. Static screening with recombinant CD81 and HEK293-derived EVs identified 1 M arginine at pH 10 as the most suitable elution condition. Dynamic chromatography on a 1 mL column using tangential flow filtration-concentrated HEK293 conditioned medium achieved 66.9% overall recovery with an elution step yield of 57.7%. In parallel, dsDNA, host cell protein, and total protein were reduced by 2 to 3 log relative to conditioned medium. Nano flow cytometry showed enrichment of the CD81-positive EV fraction from 40% in conditioned medium to more than 90% in the eluates, together with a smaller and narrower particle size distribution. These results demonstrate that Nanofitin(R)-based affinity chromatography provides a practical route toward marker-defined EV enrichment that combines selective capture, EV-compatible release, and substantial impurity clearance in a chromatography-compatible process format.

Exacerbations of respiratory viral infections significantly contribute to morbidity and healthcare burden. Among these viruses, Human Rhinoviruses (HRVs) are the most frequent causative agents of upper respiratory tract infections. To date, over 150 HRV serotypes have been identified, classified into three species: HRV-A, HRV-B, and HRV-C. No antiviral therapies are currently available against this viral family, largely due to the high serotype diversity and limited cross-protection. The major group of HRVs relies on the Intercellular Adhesion Molecule-1 (ICAM-1) receptor to infect airway epithelial cells, making ICAM-1 an attractive target for broad-spectrum therapeutic interventions. Here, we report the development of nucleic acid-based aptamers designed to disrupt ICAM-1-HRV binding and thereby prevent viral infection. Aptamers are single-stranded DNA molecules that fold into precise three-dimensional structures, enabling highly specific protein recognition. Using a Systematic Evolution of Ligands by EXponential Enrichment (SELEX) approach guided by a minimal peptide mimicking the ICAM-1 viral binding interface, a library of >1024 random single-stranded DNA sequences was screened. Through iterative rounds of selection, we identified eight candidate 77-nt DNA aptamers, which were subsequently evaluated for their potential using in silico and in vitro assays, as well as functional assays in human epithelial cells. From this strategy, two lead aptamers were selected that effectively inhibited HRV-A16 replication in a concentration-dependent manner, as measured by viral titers (TCID assay) and viral RNA quantification by RT-PCR. These findings demonstrate the potential of ICAM-1-targeting aptamers as antiviral agents capable of preventing HRV entry. By targeting a host receptor and creating a protective barrier at the cell surface, this approach may offer a broadly applicable strategy against multiple HRV serotypes, paving the way for the development of novel antiviral interventions. Graphical abstract O_FIG O_LINKSMALLFIG WIDTH=200 HEIGHT=131 SRC="FIGDIR/small/717810v1_ufig1.gif" ALT="Figure 1"> View larger version (26K): org.highwire.dtl.DTLVardef@1f0c564org.highwire.dtl.DTLVardef@2f5035org.highwire.dtl.DTLVardef@3b063eorg.highwire.dtl.DTLVardef@116ed49_HPS_FORMAT_FIGEXP M_FIG C_FIG