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Bibliography on: Metagenomics

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ESP: PubMed Auto Bibliography 02 Jan 2026 at 01:31 Created: 

Metagenomics

While genomics is the study of DNA extracted from individuals — individual cells, tissues, or organisms — metagenomics is a more recent refinement that analyzes samples of pooled DNA taken from the environment, not from an individual. Like genomics, metagenomic methods have great potential in many areas of biology, but none so much as in providing access to the hitherto invisible world of unculturable microbes, often estimated to comprise 90% or more of bacterial species and, in some ecosystems, the bulk of the biomass. A recent describes how this new science of metagenomics is beginning to reveal the secrets of our microbial world: The opportunity that stands before microbiologists today is akin to a reinvention of the microscope in the expanse of research questions it opens to investigation. Metagenomics provides a new way of examining the microbial world that not only will transform modern microbiology but has the potential to revolutionize understanding of the entire living world. In metagenomics, the power of genomic analysis is applied to entire communities of microbes, bypassing the need to isolate and culture individual bacterial community members.

Created with PubMed® Query: ( metagenomic OR metagenomics OR metagenome ) NOT pmcbook NOT ispreviousversion

Citations The Papers (from PubMed®)

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RevDate: 2025-12-31

Mehar K, K KP, Sen AP, et al (2025)

Next-Generation Eco-Omics: Integrating Microbial Function Into Predictive Ecosystem Models.

Biotechnology and applied biochemistry [Epub ahead of print].

Microorganisms drive essential ecosystem functions by mediating carbon, nitrogen, sulfur, and phosphorus transformations that regulate productivity and shape climate feedbacks. Rapid methodological advances now allow precise linkage of microbial identity, in situ activity, and ecosystem processes across spatial and temporal scales. High-resolution approaches-including long-read metagenomics and Hi-C-generate near-complete metagenome-assembled genomes (MAGs) from diverse environments, enabling reconstruction of microbial and viral-host interaction networks. Activity-resolved tools such as quantitative stable isotope probing (qSIP) and bioorthogonal non-canonical amino acid tagging (BONCAT), combined with fluorescence-activated cell sorting (FACS), yield taxon-specific growth and substrate assimilation rates within hours. Single-cell isotope techniques, including Raman-SIP and nanoSIMS, deliver nanometer-scale metabolic insights. Spatial meta-omics platforms, such as MetaFISH and MALDI-MSI, map metabolites alongside microbial identities with micrometer-level precision. Meanwhile, autonomous sequencing systems, including environmental sample processors and nanopore adaptive sampling, enable real-time (<24 h) ecological surveillance. Integrating these multimodal datasets into trait-based frameworks has reduced uncertainty in carbon flux predictions by nearly 20%. This review synthesizes these innovations, outlines optimized analytical pipelines, and proposes a framework for embedding eco-omics into predictive ecosystem and climate models, supporting evidence-driven management aligned with Climate Action and Life on Land.

RevDate: 2025-12-31
CmpDate: 2025-12-31

Mao X, Hu X, J Fang (2025)

Gut microbiota-metabolite interactions in drug-induced liver injury: mechanisms, biomarkers, and therapeutic perspectives.

Frontiers in cellular and infection microbiology, 15:1737234.

Drug-induced liver injury (DILI) remains a major obstacle in clinical pharmacotherapy and a leading cause of acute liver failure and drug withdrawal worldwide. Conventional mechanistic models centered on hepatic xenobiotic metabolism, oxidative stress, and immune injury cannot fully account for the substantial interindividual variability and the unpredictable nature of idiosyncratic DILI. Increasing evidence shows that the gut microbiota and its metabolites critically shape hepatic susceptibility through modulation of drug metabolism, inflammatory signaling, and intestinal barrier integrity. This review summarizes current understanding of the gut-liver axis in DILI pathogenesis, with a focus on microbial enzymes such as β-glucuronidase that reactivate detoxified drug conjugates, microbial dysbiosis that disrupts bile acid homeostasis, and depletion of short chain fatty acids and indole derivatives that normally support epithelial defenses and immunologic tolerance. Drug-specific microbial patterns are discussed, including acetaminophen, amoxicillin-clavulanate, anti-tuberculosis regimens, and immune checkpoint inhibitors. We introduce the concept of metabotype-dependent hepatotoxicity, which emphasizes that individual microbial metabolic profiles influence DILI risk. Advances in metagenomics, metabolomics, and integrative multi-omics enable the identification of microbial biomarkers and functional pathways associated with DILI susceptibility. Emerging therapeutic strategies include restoration of microbial homeostasis, selective inhibition of microbial enzymes, and supplementation of hepatoprotective metabolites. Finally, we outline key challenges and future directions toward translating microbiome-based insights into clinical prediction and precision prevention of DILI. Importantly, this review integrates microbial metabolic functions with precision hepatology concepts, highlighting how metabotype-driven variability can be leveraged for individualized DILI risk assessment.

RevDate: 2025-12-31
CmpDate: 2025-12-31

Yao X, Zou X, B Cao (2025)

The Human Respiratory Virome in Health and Disease: Interactions, Dysbiosis, and Methodological Challenges.

Advanced genetics (Hoboken, N.J.), 6(4):e00022.

The human respiratory virome is an underexplored component of the microbiome that includes diverse DNA and RNA viruses such as eukaryotic viruses, bacteriophages, and archaeal viruses. Recent advances in metagenomics have revealed the complexity and dynamic nature of the human respiratory virome, which interacts closely with the host and the bacterial microbiome to influence respiratory health and disease. In healthy individuals, the virome is characterized by low biomass and high temporal variability, with Anelloviruses predominant in the upper airways, whereas Streptococcus phages and herpesviruses are most commonly detected in the lower airways. Common respiratory viruses, such as respiratory syncytial virus, human rhinovirus, and influenza A virus, can persist after acute infection and modulate host immunity. The respiratory virome also plays a significant role in chronic respiratory diseases. Despite its importance, research on the respiratory virome is hampered by technical challenges, including low viral abundance and limited reference databases. This review summarizes current understanding of the composition and determinants of the respiratory virome in healthy individuals, describes its interactions with the host and respiratory microbiota, including the potential modulatory roles of bacteriophages, outlines virome alterations in respiratory diseases, examines methodological challenges, and highlights potential clinical applications and future research directions.

RevDate: 2025-12-31
CmpDate: 2025-12-31

Wagner ES, Oliphant K, D'Souza M, et al (2025)

Fecal Microbiome and Bile Acid Profiles Differ in Preterm Infants with Parenteral Nutrition-associated Cholestasis.

Journal of clinical and translational hepatology, 13(12):1036-1045.

BACKGROUND AND AIMS: Parenteral nutrition (PN)-associated cholestasis (PNAC) is frequently diagnosed in premature infants; however, not all PN-exposed infants develop PNAC. We propose that, in premature infants receiving PN and varying amounts of enteral feeds, differences in the gut microbiome and fecal bile acid content are associated with PNAC development. This study aimed to examine the fecal microbiome and bile acid content of premature infants on PN to determine if there is a relationship with the development of PNAC.

METHODS: Twenty-two preterm infants had serial bilirubin measurements and fecal samples collected during their neonatal intensive care unit admission. Fecal samples underwent 16S rRNA gene sequencing and bile acid analysis. Binomial regression, adjusting for postmenstrual age with feed amount as a moderator, was used to assess the impact of the fecal microbiome and bile acids on PNAC development.

RESULTS: Cholestatic patients (n = 11) had greater PN and antibiotic exposure (p = 0.020; p = 0.010) and longer neonatal intensive care unit stays (p = 0.0038) than non-cholestatic patients. Microbiome richness was higher in non-cholestatic infants (p < 2E-16), with no difference in β diversity (p = 1.0). Cholestatic infants had a significantly higher abundance of Proteobacteria and Fusobacteriota and a lower abundance of Bacteroidota (p < 2E-16). Akkermansia was abundant in all infants on low feeds; as feed volume increased, Akkermansia abundance significantly increased in non-cholestatic infants (p < 2E-16). Bile acid analysis demonstrated significantly lower deoxycholic acid concentrations in cholestatic infants (p < 2E-16). Metagenomic analysis revealed an increase in Proteobacteria requiring augmented stress responses in non-cholestatic infants.

CONCLUSIONS: This is the first study to directly explore the relationship between PNAC susceptibility, the microbiome, and fecal bile acids in preterm infants. The microbiome and bile acid patterns identified here may inform the development of targeted therapeutics for this vulnerable population.

RevDate: 2025-12-31
CmpDate: 2025-12-31

Wu D, Wang AJ, Bu DC, et al (2025)

The interplay between tissue-resident microbiome and host proteins by integrated multi-omics during progression of colorectal adenoma to carcinoma.

iMeta, 4(6):e70090.

The intratumoral microbiome is an emerging hallmark of cancer, yet its multi-kingdom host-microbiome ecosystem in colorectal cancer (CRC) remains poorly characterized. Here, we conducted an integrated analysis using deep shotgun metagenomics and proteomics on 185 tissue samples, including adenoma (A), paired tumor (T), and para-tumor (P). We identified 4057 bacterial, 61 fungal, 108 archaeal, and 374 viral species in tissues and revealed distinct intratumor microbiota dysbiosis, indicating a CRC-specific multi-kingdom microbial ecosystem. Proteomic profiling uncovered four CRC subtypes (C1-C4), each with unique clinical prognoses and molecular signatures. We further discovered that host-microbiome interactions are dynamically reorganized during carcinogenesis, where different microbial taxa converge on common host pathways through distinct proteins. Leveraging this interplay, we identified 14 multi-kingdom microbial and 8 protein markers that strongly distinguished A from T samples (area under the receiver operating characteristic curve (AUROC) = 0.962), with external validation in two independent datasets (AUROC = 0.920 and 0.735). Moreover, we constructed an early- versus advanced-stage classifier using 8 microbial and 4 protein markers, which demonstrated high diagnostic accuracy (AUROC = 0.926) and was validated externally (AUROC = 0.659-0.744). Functional validation in patient-derived organoids and murine allograft models confirmed that enterotoxigenic Bacteroides fragilis and Fusobacterium nucleatum promoted tumor growth by activating Wnt/β-catenin and NF-κB signaling pathways, corroborating the functional potential of these biomarkers. Together, these findings reveal dynamic host-microbiome interactions at the protein level, tracing the transition from adenoma to carcinoma and offering potential diagnostic and therapeutic targets for CRC.

RevDate: 2025-12-31
CmpDate: 2025-12-31

Liu J, Xing W, Zhang X, et al (2025)

Metagenomics and digital cell modeling facilitate targeted high-throughput sorting of anaerobic hydrogen-producing microorganisms.

iMeta, 4(6):e70082.

This study proposes a novel strategy that prioritizes functional recognition, followed by targeted high-throughput sorting, to enable the comprehensive, rapid, and efficient acquisition of target microorganisms. Using metagenomic sequencing and binning analysis, we identified 215 potential anaerobic hydrogen-producing strains from 12 large-scale biogas samples. Digital cell models were subsequently constructed from metagenome-assembled genomes, which guided the design of 14 selective culture media for enriching these hydrogen-producing bacteria. Flow cytometry-based high-throughput sorting successfully isolated 81 potential anaerobic hydrogen-producing strains, achieving a target acquisition rate above 37% and a survival rate exceeding 70%. This method holds broad potential for the discovery and sorting of functional microorganisms across diverse environments and may ultimately facilitate the development of synthetic microbiomes for industrial applications.

RevDate: 2025-12-31
CmpDate: 2025-12-31

Xiao Z, Ding K, Guo X, et al (2025)

Soil-borne legacy facilitates the dissemination of antibiotic resistance genes in soil-plant continua.

iMeta, 4(6):e70094.

Antimicrobial resistance (AMR) disseminates throughout the soil-plant continuum via complex microbial interactions. Plants shape root- and leaf-associated microbiomes that sustain plant health; however, soil-borne legacies-enriched with antibiotic-producing microbes and resistance genes-govern AMR dynamics across agroecosystems. Using 16S rRNA gene sequencing, shotgun metagenomics, and high-throughput quantitative PCR, we profiled antibiotic resistance genes (ARGs), mobile genetic elements, and virulence factor genes across bulk soil, rhizosphere, phyllosphere, and root endosphere within soil-tomato and soil-strawberry continua. Recurrent bacterial wilt amplified the resistome, particularly polypeptide resistance genes, thereby establishing the rhizosphere as a major hotspot of ARG accumulation. Multidrug-resistant Ralstonia solanacearum (R. solanacearum) strains acted as major ARG reservoirs, harboring resistance determinants on both chromosomes and megaplasmids. Collectively, these findings demonstrate that pathogen-driven restructuring of the plant microbiome accelerates ARG dissemination, establishing soil-borne diseases as critical amplifiers of AMR across agricultural ecosystems.

RevDate: 2025-12-31
CmpDate: 2025-12-31

Wang G, Zhao L, Shi Y, et al (2025)

High-throughput generic single-entity sequencing using droplet microfluidics.

iMeta, 4(6):e70087.

Single-cell sequencing has revolutionized our understanding of cellular heterogeneity by providing a micro-level perspective in the past decade. While heterogeneity is fundamental to diverse biological communities, existing platforms are primarily designed for eukaryotic cells, leaving significant gaps in the study of other single biological entities, such as viruses and bacteria. Current methodologies for single-entity sequencing remain limited by low throughput, inefficient lysis, and highly fragmented genomes. Here, we present the Generic Single-Entity Sequencing (GSE-Seq), a versatile and high-throughput framework that overcomes key limitations in single-entity sequencing through an integrated workflow. GSE-Seq combines (1) one-step generation of massive barcodes, (2) degradable hydrogel-based in situ sample processing and whole genome amplification, (3) integrated in-droplet library preparation, and (4) long-read sequencing. We applied GSE-Seq to profile viral communities from human fecal and marine sediment samples, generating thousands of high-quality single-entity genomes and revealing that most are novel. GSE-Seq identified not only dsDNA and ssDNA viruses, but also hard-to-detect giant viruses and crAssphages. GSE-Seq of bacterial genomes also revealed putative novel bacterial species, validating the versatility of this platform across different microbial kingdoms. Collectively, GSE-Seq represents a robust framework that addresses persistent challenges in high-throughput profiling for generic applications and holds immense promise for single-cell deconvolution of diverse biological entities.

RevDate: 2025-12-31
CmpDate: 2025-12-31

Lan B, Liang Y, Zhou Z, et al (2025)

Gut microbiome dysbiosis implicates the gut-bone axis in Modic changes: a metagenomic case-control study.

Frontiers in microbiology, 16:1702357.

INTRODUCTION: Modic changes (MCs) are vertebral endplate lesions strongly associated with discogenic low back pain (LBP), though their pathogenesis remains poorly understood. Emerging evidence implicates gut microbial dysbiosis in systemic inflammation and musculoskeletal disorders, yet its potential role in MCs has not been investigated. This study aimed to characterize the gut microbiome in patients with MCs and identify microbial and metabolic features linked to disease severity.

METHODS: In a case-control study, shotgun metagenomic sequencing was performed on fecal samples from 31 patients with MCs (16 Type 1, 15 Type 2) and 25 age- and sex-matched healthy controls. Microbial community structure was assessed via alpha and beta diversity analyses. Differential taxa and predictive biomarkers were identified using linear discriminant analysis effect size (LEfSe) and Random Forest modeling. Functional potential was evaluated via Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway analysis. Associations between microbial features and clinical markers (C-reactive protein [CRP], Pfirrmann grade) were also examined.

RESULTS: Patients with MCs showed significantly reduced gut microbial alpha diversity compared to controls (Chao1 index: p = 0.005; Shannon index: p = 0.034; Simpson index: p = 0.042), with the most pronounced reduction in Type 1 MCs. Beta diversity analysis revealed distinct microbial communities between groups (PERMANOVA, p = 0.001). Key discriminative taxa included unclassified_Parabacteroides (AUC = 0.895) and Bacteroides uniformis (AUC = 0.889). Metabolic pathway analysis identified 52 differentially abundant pathways, with significant enrichment of quorum sensing (p < 0.001) and glycerolipid metabolism (p < 0.001) in MC patients, both strongly correlated with elevated CRP and higher Pfirrmann grade (p < 0.001).

DISCUSSION: Gut microbial dysbiosis is associated with MCs, marked by reduced diversity, specific bacterial biomarkers, and altered metabolic pathways related to inflammation and tissue degeneration. These results suggest a potential role of the gut-bone axis in MC pathogenesis and highlight novel targets for diagnostic and therapeutic strategies in LBP.

RevDate: 2025-12-31

Xu ST, Qian ZY, Y Tao (2025)

Chinese Expert Consensus on the Application of Metagenomic Sequencing Technology in Ocular Infectious Diseases: A Delphi Method.

Biomedical and environmental sciences : BES, 38(12):1550-1557.

RevDate: 2025-12-31
CmpDate: 2025-12-31

Yagi K, Ethridge AD, Asai N, et al (2025)

Changes in Microbiome Correspond with Diminished Lung Pathophysiology Following Early-Life Respiratory Syncytial Virus Infection or Antibiotic Treatment: Microbiome Following RSV Infection.

Viruses, 17(12):.

Early-life respiratory syncytial virus (EL-RSV) infection has been implicated in long-term pulmonary disease in children. In these studies, neonatal BALB/c mice were infected at day 7 of life, leading to >35% losses in critical lung function, airway mucus metaplasia, and transcriptional hallmarks of mucus hypersecretion four weeks after RSV infection. While EL-RSV minimally reshaped the resident lung microbiota, it led to significant gut dysbiosis, including a long-term reduction of Proteobacteria that can be a source of protective metabolites related to barrier and immune function. Subsequent studies assessing whether a common infant antibiotic (ampicillin) could mitigate EL-RSV-induced lung alterations revealed further severe gut microbiome alterations and, on its own, later in life, recapitulated the full spectrum of RSV-associated alterations in lung function. Metagenomic inference showed that both RSV and ampicillin administered during early life reduced biosynthetic pathways for microbiome-derived metabolites, which are known to reinforce tight junctions, regulate inflammation, and preserve extracellular matrix elasticity. The shared loss of these metabolic programs provides a mechanistic bridge linking distinct early-life exposures to the microbiome changes and airway mechanical deficits later in life. Collectively, the data suggest that RSV and/or antibiotic-triggered gut dysbiosis is the primary insult that likely promotes improper lung maturation/repair through a metabolite-mediated mechanism and may suggest metabolite restoration as a strategy to promote proper developmental lung function.

RevDate: 2025-12-31
CmpDate: 2025-12-31

Mandal E, Noirungsee N, Disayathanoowat T, et al (2025)

TSWV Infection Differentially Reshapes the Symbiotic Microbiome of Two Frankliniella Thrips Species.

Viruses, 17(12):.

Vectoring tomato spotted wilt virus (TSWV) by two well-known thrips species, Frankliniella occidentalis Pergande and F. intonsa Trybom (Thysanoptera: Thripidae), is facilitated in different ways. Symbiotic bacteria positively influence thrips fitness, but the interaction between these bacteria and tospovirus inside the thrips' body remains unknown. Metagenomic profiling of symbionts in nonviruliferous and viruliferous Frankliniella thrips was performed to elucidate the interactions between symbiotic bacteria and the virus. A total of 97 operational taxonomic units (OTUs) were identified by profiling the microbes, where Proteobacteria was the most abundant phylum, with a high richness in Serratia spp. F. occidentalis showed lower variation in bacterial diversity between nonviruliferous and viruliferous treatments than F. intonsa. RT-qPCR validation for Serratia and Escherichia revealed opposite abundance patterns between the two thrips species. In contrast, Enterobacteriaceae and Pantoea showed similar patterns with higher abundance in nonviruliferous conditions. Wolbachia was detected exclusively in F. intonsa, with a higher bacterial titer in the viruliferous sample. Our findings suggest that TSWV association may influence the abundance of different bacterial symbionts within the thrips' body, potentially via induction of antimicrobial peptides in response to viral invasion, and to our knowledge this is the first report addressing this tripartite interaction. These findings improve our understanding of how virus-symbiont association contributes to thrips vector competence.

RevDate: 2025-12-31
CmpDate: 2025-12-31

Dos Santos MM, Salles TS, de Campos GM, et al (2025)

Acute Respiratory Infections (ARIs): Current Etiological Perspectives and Advances in Viral Metagenomics-A Review.

Viruses, 17(12): pii:v17121554.

Acute respiratory infections (ARIs) remain a leading cause of global morbidity and mortality, disproportionately affecting vulnerable populations such as children, the elderly, and immunocompromised individuals. Despite the availability of traditional diagnostic tools, including viral culture and highly elaborated PCR respiratory panels, many cases of ARI remain without an identified etiological agent. This is due to the vast diversity of viral agents that can be involved in cases of ARI, which represents a major limitation of the targeted diagnosis. In this context, viral metagenomics has emerged as a powerful, unbiased approach for detecting both known and novel pathogens directly from clinical samples. This review highlights the application of metagenomic next-generation sequencing for the investigation of etiological causes of ARIs, emphasizing its relevance in complex cases-particularly among immunocompromised patients-where standard methods might fail. We highlight the main viruses involved in respiratory infections, the strengths and limitations of metagenomic next-generation sequencing approaches, their role in genomic surveillance of respiratory viruses, and their potential to build public health responses to potentially emerging respiratory threats. Ultimately, integrating viral metagenomics into clinical and surveillance frameworks could enhance the early detection and control of respiratory viral diseases worldwide.

RevDate: 2025-12-31
CmpDate: 2025-12-31

Galeeva J, Kuzmichenko P, Manolov A, et al (2025)

Bioinformatics Tools and Approaches for Virus Discovery in Genomic Data: A Systematic Review.

Viruses, 17(12): pii:v17121538.

The exponential growth of viral metagenomic data has created an urgent need for accurate and scalable tools for virus discovery, yet the extreme diversity, rapid evolution, and limited reference databases for viruses pose unique computational challenges that traditional sequence comparison methods struggle to address. This systematic review, conducted in accordance with PRISMA 2020, examines current trends and methodological advances in virus discovery tools from 1990 to 2025. As virus discovery is a broad and multi-dimensional topic, this review focuses on the first-line tools used to analyze the results of high-throughput sequencing. The review was conducted using the PubMed database with a snowballing approach, with over 54 key studies selected for the analysis. These studies encompass the following approaches: alignment-based methods, rapid similarity estimation techniques, profile hidden Markov model methods, combination pipelines, k-mer-based approaches, and machine learning-based methods. The transition from alignment-based to machine learning methods has dramatically improved the detection of divergent viruses, yet challenges remain in interpreting model decisions and handling incomplete viral genomes. This review summarizes current knowledge and potential future directions for the development of virus detection capabilities.

RevDate: 2025-12-31
CmpDate: 2025-12-31

Lapshina VK, Guskova NI, Stetsenko IF, et al (2025)

Characterizing the Bat Virome of Vietnam: A Systematic Review of Viral Diversity and Zoonotic Potential.

Viruses, 17(12): pii:v17121532.

Bats have been identified as reservoir hosts for an exceptional diversity of viruses, including multiple taxa of high zoonotic concern. Over a hundred bat species inhabit Vietnam, which, combined with significant biodiversity, carry high risk of zoonotic spillover due to dense human-animal interfaces, extensive wildlife trade, and proximity to recent outbreak epicenters. This review systematically synthesizes data on the bat virome in Vietnam and neighboring Southeast Asian countries, assessing viral diversity, host species involvement, and zoonotic potential. By prioritizing virus groups with established zoonotic capacity and pandemic potential, the systematic search identified studies reporting viruses from 32 families across 13 bat families. Based on the WHO 2024 risk classification, seven of these viral families were categorized as high-risk, three as medium-risk, and twelve as low-risk. The comparatively higher viral diversity reported in neighboring countries suggests that the current study likely represents an underestimation of the true virome present in Vietnamese bat populations. We emphasize the urgent need for expanded virological studies integrating metagenomic sequencing, serological surveys, and ecological modeling to improve early detection of emerging threats, as the comparatively higher viral diversity reported in neighboring countries suggests existing research likely represents an underestimation of the true virome present in Vietnamese bat populations. Strengthening regional collaboration is critical for establishing proactive pandemic prevention strategies in this high-risk zoonotic hotspot.

RevDate: 2025-12-31
CmpDate: 2025-12-31

Koutsakos M, Parry RH, M Wille (2025)

Mounting Evidence for an Expanded Host Range of Influenza B Viruses.

Viruses, 17(12): pii:v17121528.

Influenza B viruses (IBV) belong to the family of Orthomyxoviridae and circulate annually in humans causing respiratory illness. Although they are considered an exclusively human pathogen, there is evidence of IBV infections in animals, including wildlife, companion animals and livestock. In addition, metagenomic studies have identified novel orthomyxoviruses in amphibians and fish that appear related to IBV, suggesting influenza viruses, including IBV, have been associated with vertebrates across their evolutionary history. In this review, we summarise our current knowledge of potential IBV and IBV-like infections in animals. These collectively suggest that the ecology of IBV extends beyond humans and warrants further investigations.

RevDate: 2025-12-31
CmpDate: 2025-12-31

Cheng H, Chong H, Yuan M, et al (2025)

Soil Microbial Life History Strategies Drive Microbial Carbon Use Efficiency Following Afforestation.

Microorganisms, 13(12):.

Soil microbial carbon use efficiency (CUE) is the core of the soil carbon (C) cycle that captures a dual microbial control point between soil organic C (SOC) accumulation and loss. The interpretation of these patterns and drivers of microbial CUE after long-term afforestation remains, however, a major scientific challenge. In particular, there are major uncertainties about the role of microbial traits in driving CUE. Here, we compared sites along a 45-year afforestation chronosequence and combined the novel [18]O-H2O tracer method with metagenomic analysis to quantify CUE and explore the mechanisms underlying microbe-mediated C dynamics. The results showed that soil microbial CUE significantly increased following afforestation and showed a positive relationship with SOC, which suggested that microbial CUE could promote C accumulation in afforested ecosystems. We further found the critical role of microbial traits in the regulation of CUE through altering microbial life history strategies: microbial CUE was positively and significantly correlated with resource acquisition (A) genes, but showed a negative and significant correlation with stress tolerance (S) strategy genes. These results suggested that soil microbes reduce investment in S strategies and shift to A and high yield (Y) strategies, thereby increasing CUE. This knowledge is important because it advances our understanding of the microbial physiological and evolutionary tradeoffs mediating soil C cycling in the context of human-induced land use change.

RevDate: 2025-12-31
CmpDate: 2025-12-31

Yamaguchi J, Orf GS, Malinauskas J, et al (2025)

teamNGS Balances Sensitivity for Viruses with Comprehensive Microbial Detection in Clinical Specimens.

Microorganisms, 13(12):.

Probe-based capture represents a highly sensitive and cost-effective approach for overcoming host background and enriching viruses in metagenomic NGS (mNGS) libraries. Using clinical specimens collected globally from patients with fever or respiratory illness, we generated mNGS libraries by random priming and Nextera XT tagmentation, followed by target enrichment (teNGS) with Comprehensive Viral Research Panel (CVRP) probes. Capture pool sizes and total reads were optimized, and libraries were initially sequenced separately. Using only 3-4% of reads required for standard mNGS, teNGS achieved increased sensitivity, 100-10,000× increases in depth, and >50% genome coverage for pathogens with titers ≥ 1000 cp/mL. Application to >2000 clinical specimens from various matrices and to contrived samples containing viruses absent from the CVRP probe set enabled detection of diverse viral families and established a minimum 65% nucleotide identity for hybridization, respectively. To save time and resources, teNGS and mNGS libraries were then combined into one sequencing run: teamNGS. In addition to streamlining the workflow, teamNGS also improved genome recovery. Coupling methods maintain the sensitivity and coverage for viruses achieved by enrichment alone while also ensuring comprehensive recovery of non-viral microbes. teamNGS has the potential to improve patient management and lower the rates of unnecessary testing and antibiotic use.

RevDate: 2025-12-31
CmpDate: 2025-12-31

Gladkov GV, Kimeklis AK, Orlova OV, et al (2025)

Taxonomic and Metagenomic Survey of a Peat-Based Straw Degrading Biofertilizer.

Microorganisms, 13(12):.

The mobilization of complex microbial communities from natural resources can be a valuable alternative to the use of single-species biofertilizers when it comes to the decomposition of plant residues. However, the functioning and interaction of microorganisms within these communities remain largely unexplored. Our task was to investigate the cellulose-degrading community using the biofertilizer BAGS (peat-based compost with straw) as an example and define its active component. For this, we monitored the succession of the biofertilizer's taxonomic composition during two consecutive rounds of its six-month composting process, varying in the applied mineral fertilization. The amount of added nitrogen significantly affected the performance of the biofertilizer, contributing to its high cellulolytic activity. Based on the network analysis, the biofertilizer's mature phase was determined, and its characteristic ASVs (amplicon sequence variants) were described. Metagenomic analysis of this phase revealed MAGs (metagenome-assembled genomes) corresponding to these ASVs, which contained genes for cellulose and aromatics degradation, as well as genes for nitrogen and sulfur pathways, including anaerobic nitrate reduction and thiosulfate oxidation. Thus, we propose that the cellulose-decomposing bacterial component of BAGS, associated with the mature phase, occupied different trophic niches, not limited to cellulose degradation, which should be considered when designing natural or artificial microbial systems for the decomposition of plant residues.

RevDate: 2025-12-31
CmpDate: 2025-12-31

Li M, Chen P, Liu C, et al (2025)

Gut Microbiome and Metabolome Signatures Associated with Heat Tolerance in Dairy Cows.

Microorganisms, 13(12):.

Heat stress significantly impairs dairy cow health and productivity, highlighting the need to understand the gut microbiome-metabolite interactions that contribute to heat tolerance. Here, we integrated metagenomic sequencing and untargeted metabolomics in twelve holstein cows selected from a previously phenotyped herd of 120 individuals, including six heat-tolerant (HT) and six heat-sensitive (HS) cows identified using entropy-weighted TOPSIS scoring. HT cows were enriched in genera such as Faecalimonas and UBA737, which were functionally linked to pathways of energy and lipid metabolism, whereas, HS cows harbored taxa associated with bacterial lipopolysaccharide and glycosphingolipid biosynthesis. A total of 135 metabolites were differentially abundant between groups. Among them, glycerol 2-phosphate and 24(28)-dehydroergosterol showed perfect classification performance (AUC = 1.000), and were mainly involved in membrane lipid remodeling and redox regulation. Integrated analysis revealed coordinated microbial-metabolite networks, exemplified by the Faecalimonas-LysoPS (16:0/0:0) and UBA737-Glycerol 2-phosphate axes, suggesting functional coupling between microbial composition and metabolic adaptation. Together, these findings demonstrate that HT cows harbor gut microbiota and metabolites favoring energy balance, membrane remodeling, and oxidative stress resilience, while HS cows display stress-related metabolic patterns. This study elucidates the microbial-metabolic mechanisms underlying thermal resilience and highlights potential biomarkers and metabolic pathways that could be applied in heat-tolerance breeding and precision management of dairy cattle.

RevDate: 2025-12-31
CmpDate: 2025-12-31

Dhakal R, Guo W, Vieira RAM, et al (2025)

Advances in Lignocellulose-Degrading Enzyme Discovery from Anaerobic Rumen Fungi.

Microorganisms, 13(12):.

Anaerobic fungi (phylum Neocallimastigomycota) play a crucial role in degrading forages and fibrous foods in the gastrointestinal tract of mammalian herbivores, particularly ruminants. Currently, they are classified into twenty-two genera; however, recent research suggests the occurrence of several novel taxa that require further characterization. Anaerobic rumen fungi play a pivotal role in lignocellulose degradation due to their unique enzymatic capabilities. This review explores the enzymatic systems of rumen anaerobic fungi, highlighting their ability to produce a diverse array of carbohydrate-active enzymes (CAZymes), such as cellulases, hemicellulases, and pectinases. These enzymes facilitate the breakdown of complex plant polymers, making anaerobic fungi essential contributors to fiber degradation in the rumen ecosystem and valuable resources for biotechnological applications. This review summarizes the structural and functional diversity of fungal CAZymes, and the mechanical disruption of plant cell walls by fungal rhizoidal networks is discussed, showcasing the ability of fungi to enhance substrate accessibility and facilitate microbial colonization. Recent studies using genomic, transcriptomic, and biochemical approaches have uncovered several novel CAZymes in anaerobic fungi, including multifunctional xylanases, β-glucosidases, and esterases. These findings highlight the continued expansion of fungal enzyme repertoires and their potential for biotechnology and feed applications. Continued research in this field will enhance our understanding of microbial ecology and enzyme function, paving the way for applications that address global challenges in energy, food security, and environmental sustainability.

RevDate: 2025-12-31
CmpDate: 2025-12-31

Li H, Yang K, Liu H, et al (2025)

Effects of Long-Term Heavy Metal Exposure on Oral Microbial Antibiotic Resistance Genes of Residents in the Mining and Smelting Area.

Microorganisms, 13(12):.

Growing evidence highlights the role of heavy metals in driving the co-selection of an-tibiotic resistance genes (ARGs), and the human oral cavity is an important reservoir of ARGs. This cross-sectional study investigated the effects of heavy metal exposure on human oral microbiota and ARGs, collecting buccal mucosal and blood samples from residents in a heavy metal-contaminated area (Baiyin City) and a non-contaminated area (Yuzhong County, Lanzhou City). The results showed heavy metal exposure is associated with altered alpha and beta diversity of bacteria and ARGs in human oral cavities, with bacterial compositional shifts being the main factor in ARG variation. Metagenomic analysis revealed heavy metal exposure is linked to modifying the interactions in the bacterial community and between ARGs and metal resistance genes (MRGs), shown by simplified topological structures in bacterial and resistome networks, along with enhanced positive correlations among nodes. Neisseria, Haemophilus, Morococcus, Streptococcus, Staphylococcus, and Mycobacteroides as potential hosts for resistance genes in human oral cavity. Furthermore, blood metal quantification revealed distinct associations with resistance patterns. This study demonstrates significant associations between environmental heavy metal exposure and the oral resistome and emphasizes the role of bacterial community composition.

RevDate: 2025-12-31
CmpDate: 2025-12-31

Cheng H, Yuan M, Ren C, et al (2025)

Soil Nitrogen Mineralization Is Driven by Functional Microbiomes Across a North-South Forest in China.

Microorganisms, 13(12):.

Nitrogen (N) mineralization is a complex microbial-driven process that controls the supply of N for plants and microbes. The relative contribution of different microbial N-cycling species/genes to the variation in N mineralization rate (NMR) across contrasting forest biomes was unclear. Here, we investigate the linkages between soil metagenomes and N mineralization rates across 10 contrasting forest biomes (covering temperate, subtropical, and tropical forests) along a 3425 km north-south forest in China. We found that the NMR was higher in subtropical forests, and the variation in NMR can be explained by climate and soil environments, particularly for soil substrate NH4[+]. Similar to NMR, microbial N-cycling genes/species were also higher in subtropical forests, suggesting that the higher microbial N-cycling traits in warm regions may drive higher NMR. We also quantified the contribution of microbial N-cycling gene pathways to NMR across forest biomes and found that the microbial N-denitrification pathway (genes like norZ, narG, nirK, and norB) and nitrification pathway (genes like nxr) explained more variation in NMR than other pathways, such as N ammonification. Collectively, our work demonstrates the importance of microbial N-cycling traits to explain soil N mineralization rates across forest biomes and suggests that this information can be used to help improve the management of the N cycle in forests across biomes.

RevDate: 2025-12-31
CmpDate: 2025-12-31

Pang Z, Guo J, Xu H, et al (2025)

Inter-Row Grassing Reshapes Nitrogen Cycling in Peach Orchards by Influencing Microbial Pathways in the Rhizosphere.

Microorganisms, 13(12):.

Traditional clean tillage in peach orchards leads to soil degradation and nitrogen (N) loss. While inter-row grassing can optimize N cycling, the specific rhizosphere microbial mechanisms involved have not been fully understood. This study investigated how different inter-row grassing modes influence N availability through microbial communities in a peach orchard. The experiment included a monoculture of Trifolium repens L. (Tr), a monoculture of Lolium perenne L. (Pr), their mixture (TPr), and clean tillage (CK). By combining soil physicochemical analyses, metagenomic sequencing, functional gene quantification, and multivariate statistics, the study systematically examined the impacts of inter-row grassing modes on soil N cycling. The results showed that inter-row grassing modes played a significant role in reshaping N processes. Pr enhanced mineralization and nitrification, increasing inorganic N through specific genes (amoA, hao). Tr, on the other hand, promoted diazotrophs (Bradyrhizobium) and dissimilatory nitrate-reducing bacteria, enhancing biological N fixation and retention. TPr combined these benefits, leading to enhanced nitrification, increased labile carbon, and elevated enzyme activities, creating a complex microbe-gene network that mediated nitrification and denitrification. Overall, inter-row grassing modulates rhizosphere functions by enhancing N cycling through a "carbon input-microbial regulation" mechanism, offering an effective strategy for improving N use efficiency and promoting sustainable orchard management.

RevDate: 2025-12-31
CmpDate: 2025-12-31

Wang C, Yu Y, Feng W, et al (2025)

The Microbial Community Succession Drives Stage-Specific Carbon Metabolic Shifts During Agaricus bisporus Fermentation: Multi-Omics Reveals CAZymes Dynamics and Lignocellulose Degradation Mechanisms.

Microorganisms, 13(12):.

This study integrates metagenomic and metabolomic data to systematically analyze the microbial community succession and carbon source metabolism transitions during the third fermentation cycle of Agaricus bisporus, with the aim of optimizing fermentation efficiency and lignocellulose degradation strategies. Principal Coordinate Analysis (PcoA) based on Bray-Curtis dissimilarity reveals significant microbial community separation across the stages of the first mushroom fruiting cycle. The overall pattern of "stage-specific differentiation" is consistent with the "cellulose hydrolysis" turn to the degradation of complex polysaccharides via carbohydrate-active enzymes (CAZymes). In the microbial network analysis showed that different microbe group controlled the stage-specific differentiation. These findings highlight a phase-dependent metabolic shift during the fermentation process: the early stages of fruiting rely more on cellulose-degrading microbes and their enzymatic systems, while later stages are driven by the degradation of complex polysaccharides and lignin derivatives, with Planctomycetota leading the degradation. This provides new mechanistic insights into agricultural waste resource utilization and the directional regulation of fermentation processes.

RevDate: 2025-12-31
CmpDate: 2025-12-31

Cui G, Cui J, Zhang M, et al (2025)

Study on the Source and Microbial Mechanisms Influencing Heavy Metals and Nutrients in a Subtropical Deep-Water Reservoir.

Microorganisms, 13(12):.

Reservoirs are hotspots for the coupling of nutrients and heavy metals, and they substantially modify the compositions and spatiotemporal distributions of microorganisms in fluvial systems. However, relatively few studies have been performed that investigate the microbial mechanisms driving interactions among heavy metals and nutrients in reservoirs. The Goupitan Reservoir, a seasonal stratified reservoir located within the Wujiang River catchment, was chosen as the research subject. The temporal and spatial variations in heavy metals and nutrients, and the metagenomic composition of the reservoir water were analyzed in January, April, July, and October 2019. The results revealed that As, Ni, Co, and Mn were derived primarily from mine wastewater, whereas Zn, Pb, Cd, and Cr were related to domestic and agricultural wastewater discharge. The study area was dominated by Proteobacteria, Actinobacteria, Cyanobacteria, and Bacteroidetes, with the proportion of dominant phyla reaching 90%. Decreases in the dissolved oxygen (DO) concentration and pH in the bottom water during July and October were conducive to increases in the abundance of the anaerobic bacterial groups Planctomycetes and Acidobacteria. The functional genes norBC and nosZ associated with denitrification (DNF), the key gene nrfAH involved in the dissimilatory nitrate reduction to ammonium (DNRA) process, the functional genes aprAB and dsrAB responsible for sulfate reduction/sulfide oxidation, as well as the thiosulfate oxidation complex enzyme system SOX, all exhibit high abundance in hypoxic water bodies and peak in the redoxcline, highlighting the significance of related nitrogen (N) and sulfur (S) metabolic processes. In addition, the concentrations of heavy metals significantly affected the spatial differentiation of the planktonic bacterial community structure, with Mn, Co, Fe, Ni, As, and Cu making relatively high individual contributions (p < 0.01). This study is important for elucidating the sources and microbiological mechanisms influencing heavy metals and nutrients in seasonally stratified subtropical reservoirs.

RevDate: 2025-12-31
CmpDate: 2025-12-31

Luo Z, Zhou Y, Wang X, et al (2025)

Effects of Endophytic Fungi and Arbuscular Mycorrhizal Fungi on Microbial Community Function and Metabolic Pathways in the Rhizosphere Soil of Festuca rubra.

Microorganisms, 13(12):.

Numerous studies have shown that there are many uncertainties associated with the interactions of nitrogen with plants and microorganisms. In particular, the effects of symbioses between plants and various microorganisms on soil microbial community function remain unclear. Metagenomic sequencing was used to explore the changes in microbial community composition, function and metabolic pathways in rhizosphere soil and the associated influencing factors under different nitrogen levels caused by arbuscular mycorrhizal fungi (AMF) inoculation of F. rubra infected with endophytic fungi and nonendophytic fungi. Plant nutrient allocation (aboveground/belowground), soil pH, and enzymatic activities significantly modulated the functional profiles of the bacterial, fungal, and archaeal communities within these rhizospheres. Soil β-glucosidase activity had the greatest effect on the cluster of orthologous groups of proteins (COG) function of the rhizosphere soil bacterial community, and soil L-leucine aminopeptidase had the greatest effect on the COG function of the rhizosphere soil fungal and archaeal communities. The contributions of AMF colonization to the kyoto encyclopedia of genes and genomes (KEGG) functions of bacteria and archaea in the rhizosphere soil were greater than those of F. rubra infection with endophytic fungi, and AMF colonization improved the metabolic pathways, secondary metabolite biosynthesis, microbial metabolism, amino acid biosynthesis and carbon metabolism of bacterial and archaeal communities in the rhizosphere soil of F. rubra. The effects of endophytic fungi and AMFs on the function and metabolic pathways of symbiotic rhizosphere soil microbial communities were heterogeneous. This study revealed that considering both biotic and abiotic factors is essential for predicting the maintenance of soil ecosystem function by plant-fungal symbionts.

RevDate: 2025-12-31
CmpDate: 2025-12-31

Han D, Zhao R, Yang X, et al (2025)

Comparative Analysis of Environmental and Host-Associated Microbiome in Odorrana schmackeri (Auran: Ranidae): Insights into Tissue-Specific Colonization and Microbial Adaptation.

Microorganisms, 13(12):.

Amphibian microbial communities are known to be shaped by host physiology and environmental factors, yet the relative roles of sexual dimorphism and tissue specialization remain poorly understood. Using 16S rRNA gene sequencing, we compared the gastrointestinal and integumentary microbiomes of a monomorphic Chinese frog population, Odorrana schmackeri, inhabiting identical montane streams. Our results showed distinct phylogenetic stratification between niches: Proteobacteria dominated both environmental microbiota and O. schmackeri gut and skin microbiotas but with differential sub-phylum specialization. The soil microbiota was dominated by unclassified_Vicinamibacteraceae, the water microbiota was Limnohabitans-dominated, the skin microbiota was dominated by Bordetella, and the gut microbiota was led by Acinetobacter. Alpha diversity analysis revealed significant tissue- and environmental-based divergences but no sexual differentiation, a pattern confirmed by beta diversity assessments showing stronger microbial community separation by tissue and environmental compartmentalization than by sex. Functional metagenomic prediction indicated convergent enrichment of metabolic pathways across host-associated and environmental communities. These results suggest that microbial community structure in O. schmackeri is principally governed by tissue-specific ecological selection pressures rather than host sexual characteristics. Our findings enhance understanding of microbiome assembly rules in vertebrate ectotherms and identify potential connections between microbiota in different ecological niches.

RevDate: 2025-12-31
CmpDate: 2025-12-31

Zhang Z, Islam MS, Noman M, et al (2025)

Enhanced Mesophilic Degradation of Rice Straw by Microbial Consortium SXJG15 Through Coordinated Enzymatic Activity and Community Reshaping.

Microorganisms, 13(12):.

The sustainable utilization of rice straw is challenged by its recalcitrant lignocellulosic structure, especially under low-to-moderate field temperatures. In this study, a novel microbial consortium (SXJG15) mainly containing Sphingobacterium, Azospirillum, and Pseudomonas was enriched from overwintering rice stubble in Zhejiang, China, and evaluated for its rice straw degradation efficiency at 25 °C. Over an 18-day cultivation period, SXJG15 achieved a 52.5% degradation of total rice straw, including 60.2% cellulose, 76.3% hemicellulose, and 40.7% lignin. High extracellular enzymatic activities, including cellulases (up to 80.3 U/mL) and xylanases (up to 324.8 U/mL), were observed during the biodegradation process. 16S rRNA gene sequencing and metagenomics analyses revealed a succession of dominant taxa, including Sphingobacterium, Azospirillum, and Cellulomonas. Further, CAZy annotation indicated that the SXJG15 enzyme system was rich in glycoside hydrolases (42.7%) and glycosyltransferases (34.2%), demonstrating its high potential for lignocellulose degradation. This study uniquely demonstrates the mesophilic (moderate temperature 25 °C) efficiency of SXJG15 in lignocellulose breakdown, provides new insights into the microbial mechanisms of straw decomposition, and lays a foundation for bioenergy and soil fertility applications for developing a sustainable agriculture system.

RevDate: 2025-12-31
CmpDate: 2025-12-31

Cui Y, Wang S, Zhao W, et al (2025)

Shared Gut Microbial and Functional Signatures Linking Parkinson's Disease and Type 2 Diabetes Revealed by Function-Anchored Metagenomics.

Microorganisms, 13(12):.

Parkinson's disease (PD) and type 2 diabetes mellitus (T2DM) exhibit increasing comorbidity, yet the shared contribution of gut microbiota remains unclear. To investigate parallel microbial and functional alterations underlying PD, T2DM, and PD with diabetes (PDDM), we performed fecal metagenomic sequencing in 156 PD, 41 T2DM, and 44 PD with diabetes (PDDM) patients and 83 healthy controls (HC). PD and T2DM showed highly concordant microbial shifts, with 22 genera and 91 species consistently altered across disease groups compared with HC. Functional enrichment highlighted common perturbations in taurine and hypotaurine metabolism, retinol metabolism, the hypoxia-inducible factor-1 (HIF-1) pathway, and xenobiotic degradation, implicating disrupted oxidative stress responses, neuro-metabolic regulation, and detoxification. Key taxa, including Limosilactobacillus fermentum, Lactobacillus porci, and Lactobacillus delbrueckii, were increased and showed moderate positive correlations (|ρ| ≥ 0.3) with antioxidant/retinol-HIF-1, taurine-hypotaurine, and xenobiotic degradation pathways. Bifidobacterium breve (unadjusted analysis) was increased in PD and further enriched in PDDM, correlating with multiple beneficial pathways. Bifidobacterium simiarum (covariate-adjusted analyses) showed the broadest positive pathway associations, while selected Bacteroides species (e.g., B. acidifaciens) exhibited negative correlations with insulin-resistance pathways and positive correlations with steroid hormone biosynthesis. By contrast, Butyricimonas vaginalis showed negative correlations with HIF-1 and insulin signaling and with cytochrome P450-related drug metabolism. These findings provide the first systematic evidence of parallel taxonomic and functional dysbiosis in PD and T2DM, supporting gut microbiota as a shared mediator and potential therapeutic target in comorbidity.

RevDate: 2025-12-31
CmpDate: 2025-12-31

Ji J, Jiang X, Song P, et al (2025)

Multi-Omics Insights into Microbial Interactions and Fermented Food Quality.

Microorganisms, 13(12):.

The quality, flavor, and functional attributes of fermented foods are intrinsically shaped by the composition and metabolic dynamics of their microbial communities. This systematic review explores the structural organization, successional patterns, and mechanistic roles of these communities in influencing food quality, with a specific focus on core functional groups-including lactic acid bacteria (LAB), yeasts, and molds-and their interplay through key metabolic pathways. By integrating multi-omics approaches, such as metagenomics and metabolomics, we elucidate the underlying relationships between microbial activity and the formation of volatile flavor compounds, nutritional metabolites, and bioactive substances. These insights offer a scientific basis for the targeted regulation and functional enhancement of fermented food products.

RevDate: 2025-12-31
CmpDate: 2025-12-31

Qi X, Han Z, Meng J, et al (2025)

Integrated Metagenomic and Lipidomic Profiling Reveals Dysregulation of Facial Skin Microbiome in Moderate Acne Vulgaris.

Microorganisms, 13(12):.

Acne vulgaris is a prevalent chronic inflammatory dermatosis primarily affecting the pilosebaceous units. Current therapeutic approaches often exhibit limited efficacy and high recurrence rates. To investigate the microbiome-related mechanisms of acne vulgaris, facial skin samples from 19 patients with moderate acne and 20 healthy individuals were analyzed using an integrated metagenomic and lipidomic profiling strategy. Metagenomic analysis revealed a significant reduction in microbial diversity (Chao index) in acne-affected skin compared to healthy controls (p < 0.001). The relative abundance of Staphylococcus, particularly Staphylococcus epidermidis, was significantly elevated in acne group (p < 0.05), while Cutibacterium acnes levels remained unchanged. Carbon metabolism pathways were enriched in the acne group (p < 0.05), predominantly driven by Cutibacterium, whereas other enriched metabolic pathways, such as ABC transporters and glycine, serine, and threonine metabolism (p < 0.05), showed a greater contribution from Staphylococcus. Virulence factors enriched in acne samples were primarily offensive in nature and largely attributed to Staphylococcus. Moreover, acne-associated microbiome exhibited a significantly higher prevalence of resistance genes against fluoroquinolones, fosfomycin, and triclosan (p < 0.05). Untargeted lipidomic analysis demonstrated significantly elevated total serum and triglyceride levels, along with a reduction in fatty acid chain length and a higher degree of saturation compared to the healthy group (p < 0.01). Specific triglycerides significantly enriched in the acne group, such as TG (15:0_14:0_16:0) + NH4, exhibited a significant positive correlation with Staphylococcus. This correlation is associated with elevated clinical erythema and melanin indices, suggesting that Staphylococcus is implicated in the development of acne-related inflammation. Additionally, Thermus exhibits negative correlations with acne-associated lipids and inflammatory parameters, potentially exerting a protective role. These findings suggest that Cutibacterium and Staphylococcus play differential yet synergistic roles in acne pathogenesis. The observed skin microbiome dysbiosis and lipid metabolic alterations provide novel insights into the pathophysiology of acne vulgaris, which may inform the development of targeted therapeutic strategies.

RevDate: 2025-12-31
CmpDate: 2025-12-31

Wang Y, Gong L, Gao Z, et al (2025)

Comparative Analysis of Sponge-Associated, Seawater, and Sediment Microbial Communities from Site F Cold Seep in the South China Sea.

Microorganisms, 13(12):.

Microbial communities at Site F cold seep, ubiquitous in both the environment and the associated fauna, demonstrate clear habitat-specific partitioning. Metagenomic sequencing and binning demonstrated a striking partitioning of microbial taxa at the cold seep: whereas the sponge-associated microbiome was distinctly enriched with specialized sulfur- and methane-oxidizing bacteria that were rare in the environment, it simultaneously exhibited a significantly reduced archaeal content, lower α-diversity, and a simpler overall community structure compared to the sediment and seawater communities. Distinct evolutionary lineages and varying abundances were observed among the microbiomes from seawater, sediment, and sponges. Furthermore, their Metagenome-Assembled Genomes (MAGs) exhibited significant differences in genomic features, including genome size and GC content. The sponge-associated microbiome exhibits lower diversity but maintains a high abundance of key functional genes, particularly those involved in sulfur cycling (e.g., apr, dsr, metZ), indicating enhanced metabolic efficiency in energy conservation and nutrient acquisition. This study reveals that the seawater, sediment, and sponge-associated microbiomes exhibit genome simplification and functional specialization in the cold seep environment, with varying lifestyles driving structural optimization and functional remodeling of the symbiotic microbiomes.

RevDate: 2025-12-31
CmpDate: 2025-12-31

Brouwir L, KleinJan H, Balent C, et al (2025)

Fate and Removal of Antibiotics and Antibiotic Resistance Genes in a Rural Wastewater Treatment Plant: A Microbial Perspective of Nature-Based Versus Advanced Technologies.

Microorganisms, 13(12):.

Antibiotics (ATBs), antibiotic-resistant bacteria (ARB), and antibiotic resistance genes (ARGs) present an increasing threat/challenge to our environment and human health, resulting in increasingly strict wastewater management regulations through the revised Urban Wastewater Treatment Directive (UWWTD, 2024/3019/EU). This study evaluates a vertical-flow constructed wetland (CW) as a nature-based solution for removing ATBs, ARB, and ARGs from urban wastewater. The CW operated alongside two conventional quaternary treatments: granular activated carbon (GAC) and ozonation combined with GAC (O3 + GAC). Hydraulic conditions were kept stable across seasonal variations. Three antibiotics were quantified (through LC-MS/MS) in parallel to ARGs (through qPCR and metagenomics) and bacterial profiling (metabarcoding and plate counts). Results indicate that under the conditions tested (rural environment; UWWTP 13.000 p.e.), the CW achieves effective and stable removal of ATBs and ARGs. This study highlights the potential of nature-based solutions to match advanced quaternary treatments in removal performance and operational reliability, offering a sustainable and cost-effective means to reduce the spread of ATBs and ARGs via wastewater.

RevDate: 2025-12-31
CmpDate: 2025-12-31

Tkaczyk M (2025)

Biofilm as a Key Element in the Bacterial Pathogenesis of Forest Trees: A Review of Mechanisms and Ecological Implications.

Microorganisms, 13(12):.

Bacterial diseases of forest trees represent an increasing threat to ecosystem health and the sustainability and resilience of forest management, particularly under changing climate conditions. One of the key yet still insufficiently understood adaptive mechanisms of pathogens is biofilm formation-a structured community of bacterial cells embedded in a matrix of extracellular polymeric substances (EPS), which provides protection against stress factors, biocides, and the host's defensive responses such as antimicrobial compounds or immune reactions. This paper presents a comprehensive review of current knowledge on the role of biofilms in the bacterial pathogenesis of forest trees, covering their formation mechanisms, molecular regulation, and ecological significance. Four key stages of biofilm development are discussed-adhesion, microcolony formation, EPS production, and dispersion-along with the roles of quorum sensing systems and c-di-GMP-based signaling in regulating these processes. Examples of major tree pathogens are presented, including Pseudomonas syringae, Erwinia amylovora, Xylella fastidiosa, the Brenneria-Gibbsiella complex associated with Acute Oak Decline (AOD) and Lonsdalea populi. Biofilm formation is shown to play a crucial role in the colonization of xylem, leaf surfaces, and tissues undergoing necrosis, where biofilms may stabilize decomposition zones and support saprophytic-pathogenic transitions. In the applied section, the concept of "biofilm-targeted control" is discussed, encompassing both chemical and biological strategies for disrupting biofilm structure-from quorum-sensing inhibitors and EPS-degrading enzymes to the use of biosurfactants and antagonistic microorganisms. The need for in situ research in forest environments and the adaptation of advanced imaging (CLSM, micro-CT) and metagenomic analyses to tree systems is also emphasized. This review concludes that biofilms are not merely a physiological form of bacterial organization but a complex adaptive system essential for the survival and virulence of pathogens in forest ecosystems. Understanding their functions is fundamental for developing sustainable and ecologically safe phytosanitary strategies for forest protection.

RevDate: 2025-12-31
CmpDate: 2025-12-31

Liu P, Wang D, Zhang F, et al (2025)

Seasonal Metagenomic Survey of Pathogenic Microorganisms in Non-Human Primates in Mayanghe National Nature Reserve, China.

Pathogens (Basel, Switzerland), 14(12): pii:pathogens14121237.

Understanding the diversity of pathogenic microorganisms in wild primates is essential for assessing their health and zoonotic risks. In this study, metagenomic sequencing was applied to investigate the composition and seasonal dynamics of potential pathogenic microorganisms in the feces of François' langurs. A total of 77 potential pathogenic taxa were identified, mainly belonging to Bacillota and Pseudomonadota. The most abundant genera were Streptococcus, Staphylococcus, Salmonella, Listeria, and Pseudomonas, while dominant species included Staphylococcus aureus, Streptococcus pneumoniae, Salmonella enterica, Listeria monocytogenes, and Escherichia coli. Significant seasonal differences were detected in both α- and β-diversity indices, with higher microbial diversity in spring and distinct community structures across seasons. Several genera and species, including Vibrio, Chlamydia, Mycobacteroides, Vibrio cholerae, Yersinia enterocolitica, Chlamydia trachomatis, and Mycobacteroides abscessus, showed marked seasonal fluctuations. The findings reveal that the pathogenic microbial community of François' langurs is strongly shaped by seasonal environmental factors. The detection of multiple zoonotic pathogens suggests a potential risk of cross-species transmission, providing valuable baseline data for primate disease ecology and conservation health management.

RevDate: 2025-12-31
CmpDate: 2025-12-31

Shuvo MSH, Kim S, Jo S, et al (2025)

Flavonifractor plautii as a Next-Generation Probiotic Enhancing the NGP F/P Index in a Simulated Human Gut Microbiome Ecosystem.

Pharmaceutics, 17(12): pii:pharmaceutics17121603.

Background/Objectives: Traditionally consumed fermented foods and lactic acid bacteria (LAB)-based products have primarily been investigated for their nutritional and health-promoting benefits as dietary supplements. More recently, research has advanced toward exploring their therapeutic potential in pharmaceutical development. However, reliance on conventional LAB strains despite their established safety and efficacy has led to saturation at the strain level, underscoring the need for next-generation probiotics (NGPs) with novel therapeutic potential. In this context, we identified Flavonifractor plautii from human feces as a candidate NGP and investigated its effects on the human gut microbiota. Methods: Whole-genome sequencing revealed distinct genetic features that supported its uniqueness, and the strain was designated PMC93. A human gut microbial ecosystem simulator was used to administer F. plautii daily for one week, after which microbial community changes were evaluated using 16S rRNA gene-based metagenomic sequencing. Results: The administration did not induce significant changes in alpha or beta diversity, suggesting that F. plautii does not disrupt overall bacterial community structure, thereby supporting its microbial community safety. Taxonomic analysis demonstrated a significant increase in the Firmicutes-to-Proteobacteria ratio (NGP F/P index). The improvement surpassed that of conventional LAB treatments and was consistently maintained under supplementation with commonly encountered pharmaceutical compounds and nutrients. The shift was associated with an increase in short-chain fatty acid (SCFA)-producing beneficial taxa and a decrease in pro-inflammatory and potentially pathogenic groups. Functional outcomes, including elevated SCFA levels and downregulation of inflammation-related gene expression, further corroborated these compositional changes. The strain also demonstrated safety in in vivo models. Conclusions: Collectively, these findings suggest that strain PMC93 is a promising NGP candidate with substantial therapeutic potential for microbiota-associated health and disease modulation, particularly due to its ability to enhance the NGP F/P index.

RevDate: 2025-12-31
CmpDate: 2025-12-31

Karpova D, Belkina D, Porotikova E, et al (2025)

Metagenomic Study of the Grapevine Decline Detected a Cocktail of Fungi Associated with Grapevine Trunk Diseases.

Plants (Basel, Switzerland), 14(24):.

This study analyzed the microbiome of three varieties differing in genotype and technical purpose: Cristal, Riesling, and Avgustin, all exhibiting decline symptoms of unknown etiology. A total of 92 symptomatic and asymptomatic grapevines were analyzed using ITS and 16S rRNA amplicon sequencing and molecular genetic methods. Phytoplasmas and the pathogenic bacteria Xylella fastidiosa and Xylophilus ampelinus were not present in the samples. The decline symptoms were associated with a cocktail of fungal pathogens that cause grapevine trunk diseases. In particular, the analysis revealed the causative agents of Botryosphaeria dieback (Sphaeropsis spp. and Botryosphaeria spp.), fungi associated with the Esca complex (Phaeomoniella spp., Phaeoacremonium spp., Inonotus spp., Seimatosporium spp., Stereum spp., and Cadophora spp.), and the causative agents of Phomopsis dieback (Diaporthe spp.). The symptoms of decline may be increased by several facultative grapevine pathogens that have been identified in microbiome (genera Stemphylium, Alternaria, Aspergillus, Penicillium, Talaromyces, and Fusarium). The metagenomic data of the grapevine microbiome provides opportunities for developing disease control strategies, which is important for the sustainable management of vineyards.

RevDate: 2025-12-30

Mbabazi M, Kateete DP, Nakazzi F, et al (2025)

The impact of tuberculosis and its treatment on the lung and gut microbiota: a global systematic review, meta-analysis, and amplicon-based metagenomic meta-analysis.

BMC infectious diseases pii:10.1186/s12879-025-12369-1 [Epub ahead of print].

RevDate: 2025-12-30

Yao L, Solania A, Luissint AC, et al (2025)

The Secreted Metabolite Isopentenyladenine from Faecalibacterium prausnitzii Is Anti-inflammatory with Barrier-Protective Properties.

ACS infectious diseases [Epub ahead of print].

Colonic microbiome dysbiosis is correlated with inflammatory bowel disease (IBD), and depletion of the commensal bacterium Faecalibacterium prausnitzii (F. prausnitzii) is routinely observed in the metagenomic analyses of IBD patient microbiome samples. F. prausnitzii is likely beneficial to hosts, as oral administration of F. prausnitzii strain A2-165 has anti-inflammatory properties in murine models of colitis. Previous studies attribute the anti-inflammatory effects of F. prausnitzii A2-165 to production of the short-chain fatty acid butyrate, as well as a secreted protein known as microbial anti-inflammatory molecule (MAM). Here, we verified that oral dosing of strain A2-165 protects against DSS-induced murine colitis and further showed that the aqueous-soluble secreted fraction of overnight cultures from a collection of F. prausnitzii strains inhibits inflammatory signatures, including the activation of the host's NF-κB pathway, production of IL-8, and differentiation of naïve T cells into the TH17 lineage. Our findings against a panel of in vitro assays suggested that the anti-inflammatory responses were attributable to secreted small-molecule or peptide metabolites, as both heat-inactivated and proteinase K-treated F. prausnitzii culture supernatants retained activity. Untargeted and targeted mass spectrometry metabolomics analyses on the soluble anti-inflammatory secretome yielded several unique F. prausnitzii metabolites, including isopentenyladenine. We demonstrated that isopentenyladenine independently modulates host cellular signaling and immune responses and suggest that this newly identified metabolite with human immunomodulatory properties may be useful toward the discovery of IBD-focused therapeutics.

RevDate: 2025-12-30

Liu X, Ma T, Khan I, et al (2025)

Age-dependent variations in aerosol-borne particulates and microbial communities in multi-tier broiler housing systems: A metagenomics environmental health risk assessment.

Poultry science, 105(2):106308 pii:S0032-5791(25)01548-2 [Epub ahead of print].

By investigating the temporal dynamics of airborne microbial communities associated with particulate matter in multi‑tier broiler housing, this study offers a systematic reference for understanding how environmental microbiota shift with broiler age. Fine particulate matter (PM2.5) and total suspended particulate (TSP) were collected from the housing environment at three growth stages: D10 (10-day-old), D24 (24-day-old), and D38 (38-day-old). The concentration and LPS content of the collected TSP and PM2.5 samples from each stage were measured, followed by metagenomic sequencing. Results revealed that the concentrations of TSP and PM2.5 peaked at D24 (P < 0.05), showing a trend of first increasing and then decreasing, and the change trend of mortality in the early stage was similar. Metagenomic results identified that Faecalibacterium, Pseudomonas, and Acinetobacter were the dominant genera at D24, whereas Enterococcus and Macrococcus were the dominant genera at D10 and D38, respectively. Correlation analysis further indicated that TSP was positively associated with mortality and g_Pseudomonas, while PM2.5 was positively associated with g_Faecalibacterium. Significant enrichment was observed in metabolic pathways such as glycosyltransferase 35 and glycoside hydrolase 23, macB, LOS(CVF494), and other antibiotic resistance and virulence genes in TSP and PM2.5 (P < 0.05). Collectively, these findings elucidate the stage‑specific dynamics of environmental microbiota in broiler housing and underscore particulate matter as a potential driver of both microbial shifts and health outcomes, thereby providing evidence to inform interventions aimed at improving environmental and flock health.

RevDate: 2025-12-30

Feng W, Ma R, Guo Y, et al (2025)

Rhizosphere metagenomics and metabolomes provide new insights into the relationship between rhizosphere microecology and early bolting of Angelica dahurica.

Microbiological research, 305:128435 pii:S0944-5013(25)00394-5 [Epub ahead of print].

Angelica dahurica is a medicinal and edible plant with a wide range of pharmaceutical and food applications. However, the early bolting, which leads to reduced yield and loss of bioactive constituents, has become a major obstacle to the industrial development of A. dahurica. Rhizosphere microecology affects plant growth and secondary metabolite accumulation, but the association of rhizosphere microecology with the early bolting of A. dahurica is not fully understood. This study integrated metagenomic and metabolomic analyses to systematically characterize the differences in rhizosphere microecology of non-bolting and early bolting A. dahurica plants. Results revealed significant disparities in soil physicochemical properties, root exudate profiles, and microbial community composition between two groups, all of which exhibited correlations with the coumarin compounds content, the primary pharmacologically active constituents of A. dahurica. Integrated analysis suggested that root-derived acyl-homoserine lactone (AHL) quorum-sensing signals, as the primary chemical signals of the prevalent Gram-negative bacteria, may participate in regulating the microbial community structure and soil properties, thereby influencing the bolting and flowering process. This study proposes a potential complex regulatory network of "rhizosphere microbiome - quorum-sensing signals - soil nitrogen cycle - bolting and flowering" linking the rhizosphere microecology to early bolting in A. dahurica, thereby addressing a key knowledge gap in this area. The findings offer a scientific foundation and innovative strategy for the simultaneous prevention of early bolting and quality improvement in A. dahurica through soil microecological management, which is of significant importance for promoting the sustainable commercial development of the A. dahurica industry.

RevDate: 2025-12-30

Yu X, Huang S, Tang J, et al (2025)

Multi-omics reveals efficient thiamethoxam biodegradation but altered flavor profile by native microbiota during Pixian broad bean paste fermentation.

International journal of food microbiology, 449:111600 pii:S0168-1605(25)00545-8 [Epub ahead of print].

Thiamethoxam (TH), a systemic neonicotinoid insecticide, poses food safety risks due to its persistence and uptake in crops. Microbial degradation during fermentation offers a promising decontamination strategy, but the underlying mechanisms and impact on food quality remain unclear. This study investigated TH and its toxic metabolite clothianidin biodegradation in Pixian broad bean paste (PBP) fermentation, assessed the impact of residue dissipation on product quality, and revealed microbial responses and metabolic adaptations under pesticide stress. Results demonstrated that TH and clothianidin were nearly completely degraded in the PBP fermentation system within 16 days, with a half-life of 3.25 days. Metagenomic analysis revealed that TH stress enriched pollutant-degrading microbes (e.g., Aspergillaceae, Desulfobacterota) and upregulated xenobiotic degradation genes (e.g., drug metabolism). However, volatile flavor compounds analysis indicated that TH treatment altered the flavor profile by reducing esters and phenols while increasing ketones and acids. Integrated metabolomics demonstrated that TH may disrupt organic acid metabolism during early fermentation, suppressing downstream flavonoid transformation and amino acid biosynthesis, ultimately compromising nutritional quality and flavor attributes. Multi-omics integration revealed that TH stress reshaped microbial community structure and enabled dual regulation of pesticide degradation and fermentation pathways through coordinated gene expression, ultimately altering PBP fermentation quality. Therefore, these findings demonstrate that the native microbial community in PBP efficiently degrades neonicotinoid pesticides, providing a novel strategy for the bioremediation of fermented foods and serving as an emerging reservoir of potential safe degrading bacteria, while highlighting the necessity for optimized microbial interventions to minimize adverse effects on product quality.

RevDate: 2025-12-30

Ha LH, On YY, Pohan C, et al (2025)

High-throughput single-cell isolation of Bifidobacterium strains from the human gut microbiome.

Microbiology spectrum [Epub ahead of print].

UNLABELLED: Bifidobacterium represents a diverse genus of commensal gut bacteria with key roles in human health, from metabolizing indigestible fibers to protecting against pathogens. While metagenomic studies have highlighted significant strain diversity for Bifidobacterium species within individuals, their systematic isolation and phenotypic characterization can be hampered by the significant effort and biases inherent in traditional culturomics. Here, we explored the utility of a high-throughput single-cell dispensing system (B.SIGHT)-based workflow for accelerating the process of isolating diverse Bifidobacterium strains from fecal samples. Systematic assessment of this workflow revealed a high single-cell dispensing frequency (>88%) and the ability to preserve species diversity when a pool of Bifidobacterium strains was dispensed. Culturing-related factors including the use of an effective selection medium, such as the Bifidus Selective Medium supplemented with mupirocin, and the length of pre-dispensing incubation were found to be critical in determining isolation success. Leveraging this workflow, we obtained a total of 622 viable isolates from five Singaporean fecal samples, of which >98% were found to be from Bifidobacterium species. Whole-genome sequencing of 96 isolates identified six different Bifidobacterium species with both inter- and intra-subject strain and lineage diversity, and the majority (>66%) were novel relative to large public genomic databases. Our findings highlight the ability of this high-throughput culturomics workflow to accelerate the recovery of diverse and novel Bifidobacterium strains, enabling further interrogation of their functional characteristics and advancing our understanding of important bacterial species in the gut microbiome.

IMPORTANCE: The field of high-throughput microbial culturomics is still in its early stages. Enhancing our ability to isolate and phenotypically test bacterial strains from complex communities is crucial for advancing microbiome research and healthcare development. Given the time and cost inefficiencies of traditional culturing methods, a more efficient, high-throughput approach to obtain isolates is needed. In the present study, we assessed a single-cell dispensing platform and developed a workflow to isolate diverse Bifidobacterium strains from fecal samples. We demonstrated here the capability of this novel technology to efficiently obtain hundreds of isolates of a targeted group, covering both species and strain diversities. This generalizable and scalable method can potentially allow for the high-throughput recovery of microbes from other taxonomic groups, providing a fundamental step in improving the culturomics framework to complement metagenomic approaches and enable isolate-level functional studies of important microbes.

RevDate: 2025-12-30

McParland EL, Wittmers F, Bolaños LM, et al (2025)

Seasonal patterns of DOM molecules are linked to microbial functions in the oligotrophic ocean.

mSystems [Epub ahead of print].

Hundreds of thousands of individual microbe-molecule interactions regulate the flux, transformation, and fate of carbon stored in the climatically important reservoir of marine dissolved organic matter (DOM). While marine microbial communities have been characterized at high resolution for over a decade, observations of the molecules cycled by the microbial-chemical network at similar resolution are limited. In addition, bulk characterizations of DOM can mask the complex network of interactions comprised of rich chemical diversities. Here, we present a three-year, depth-resolved, molecular time-series of DOM and prokaryoplankton at the Bermuda Atlantic Time-series Study (BATS) site. Both time-series exhibited seasonality that was compositionally distinct and primarily endemic to one sampling depth. We also putatively identified four exometabolites (gonyol, glucose-6-sulfate, succinate, and trehalose) that exhibit seasonal accumulation. We hypothesize these patterns result from environmental conditions that alter community composition on a seasonal timescale and thus shift the relative proportions of microbial functions that produce and consume the substrates. Critically, we observed the interannual composition of seasonal DOM molecules to be more stable than the taxonomy of the microbial community. This points to an important role of functional redundancy in regulating DOM composition. We tested this observation by querying metagenomes for pathways that utilize metabolic by-products putatively identified in the DOM time-series. We find that core microbial metabolisms, either those required by all or by a subset of marine microbes, are important predictors of DOM composition. The molecular-level characterization of DOM herein highlights the potential imprint of microbial activity on seasonal DOM composition.IMPORTANCEMarine dissolved organic matter (DOM) is a major carbon reservoir that acts as a critical control on the Earth's climate. DOM dynamics are largely regulated by a complex web of chemical-microbial interactions, but the mechanisms underpinning these processes are not well understood. In a three-year time-series, we found that the identity of the microbes is more likely to change between years than the composition of the DOM molecules. The taxonomic variability suggests that metabolisms shared across taxa, encoded by genes that conduct core microbial functions, are responsible for the more stable composition of DOM. While more than three decades of marine prokaryoplankton time-series are available, a similar reference for DOM molecules was missing. This time-series provides an improved understanding of the different responses of DOM molecules and microbes to seasonal environmental changes.

RevDate: 2025-12-30
CmpDate: 2025-12-30

Sun Y, Li P, Wang X, et al (2025)

Gut dysbiosis in early severe burns contributes to acute lung injury by impairing neutrophil chemotaxis.

Journal of leukocyte biology, 118(1):.

Severe burns complicated by acute lung injury are critical causes of respiratory failure and multiple organ dysfunction syndrome. Neutrophils extensively infiltrate lung tissues early postburn to mediate pulmonary damage, but the underlying mechanisms remain unclear. We analyzed gut microbiota of severe burn patients via metagenomics and metabolomics, assessed neutrophil chemotaxis using a self-developed in vitro agarose model, and validated Faecalibacterium prausnitzii and butyrate's effects on restoring neutrophil chemotaxis in gut microbiota-depleted mice via oral gavage (plus in vivo validation with small animal imaging). Bronchoalveolar lavage fluid biomarkers and pulmonary function tests evaluated pulmonary injury from impaired neutrophil chemotaxis. Early postburn, F. prausnitzii and its metabolite butyrate were significantly depleted in patients, concurrent with impaired neutrophil chemotaxis-restored by butyrate supplementation. In murine burn models, F. prausnitzii or butyrate rescued neutrophil chemotaxis, reduced pulmonary neutrophil infiltration, and attenuated lung injury. Mechanistically, butyrate restored neutrophil function in a severe burn patient plasma-stimulated model by downregulating P2X1 receptor expression and suppressing myosin light chain phosphorylation. Our findings indicate postburn gut microbiota dysbiosis and metabolite alterations disrupt neutrophil chemotaxis, causing excessive pulmonary neutrophil infiltration/activation. This highlights gut microbiota-derived metabolites as potential therapeutics for mitigating neutrophil-driven lung injury early postsevere burns.

RevDate: 2025-12-30
CmpDate: 2025-12-30

Kulohoma BW, I Ng'eno (2022)

NOMAD: metagenomic characterisation of the viral pathogen composition in outbreaks of non-malaria acute febrile illness cases.

Open research Africa, 5:24.

The clinical importance of non-malaria febrile acute illness (NM-AFI) in patients with a negative parasitological test has become apparent, with the progressive reduction in malaria transmission in endemic regions. Bacterial pathogens, for example Streptococcus pneumoniae and Haemophilus influenzae, which contribute disproportionally to febrile illness, are now preventable by vaccines. However, there are no vaccines, and little is known about viral NM-AFI prevalence, proliferation, virulence, and transmission chains between hosts. Although the predominant viral causes of NM-AFI are established, it is unclear if there are other NM-AFI associated emerging infectious viral pathogens that previously remained undetectable by conventional diagnostic strategies, for example severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2). Presumptive broad-spectrum antibiotic prescriptions to aparasitaemic patients not only drive drug resistance, but also lead to poor treatment outcomes. We hypothesized that insights on NM-AFI etiology, and consequently case management, could be improved by exploiting viral sequence diversity to identify viral pathogens present within metagenomics samples. We exploited simulated and existing infectious disease (Ebola, hepatitis C, chikungunya, and mosquito-borne arboviruses) metagenomic datasets to determine the composition of viral pathogens present, by implementing profile Hidden Markov Models derived from Swiss-Prot viral reference sequences for accurate pathogen detection and classification. Our analysis identified a combination of sequences from multiple viral etiological agents within the same disease sample. This approach provides a granular perspective of multiple viral etiological agents present within a single intra-host disease episode. It highlights prevalent viral strains that can subsequently be routinely detected using directed diagnostic tests to improve disease surveillance in endemic regions.

RevDate: 2025-12-30
CmpDate: 2025-12-30

Sharma S, Gupta N, Kaur S, et al (2025)

Soil microbial resources: Unlocking sustainable strategies for crop productivity and soil health.

Current research in microbial sciences, 9:100468.

Using efficient rhizospheric microbes as bio-inoculants is a key factor in promoting agricultural sustainability, as these microbes have been shown to enhance plant growth promotion and crop productivity. Soil microbial communities offer numerous benefits to crops, including improved nutrient uptake efficiency, phytohormone production, improved soil structure, nutrient balance, enhanced plant and soil health, protection against soil borne phyto-pathogens and increased plant resilience to abiotic and biotic stresses. These characteristics of rhizospheric microbes have attracted researcher's attention, as using these microbes individually or in consortia has enhanced soil fertility as well as soil heatlh in a eco-friendly manner. Further, the utilization of multi-omics techniques for exploring the hidden potential of beneficial plant growth promoting microbes is one of the novel approaches that can result in the generation of new biological formulations under changing climatic conditions. This review is therefore focused on the diversity of microbes, their contributions for plant growth promotion and yield with advances in techniques for screening and development of efficient bio-inoculant with a special emphasis on promoting sustainable agriculture practices.

RevDate: 2025-12-30
CmpDate: 2025-12-30

Fernandes P, Cano-Díaz C, Pinto R, et al (2025)

The composition and structure of bacterial and fungal communities in kiwifruit are influenced by photoselective nets.

Current research in microbial sciences, 9:100424.

Photoselective nets in agriculture are typically designed to modify the light spectrum, intensity, and microclimate around crops, influencing plant growth, productivity, and quality. However, knowledge regarding their impact on the microbiota of plants and fruits remains limited. This study assessed the impact of pearl, grey, and yellow photoselective nets on the microbial communities present on kiwifruit surfaces using amplicon high-throughput sequencing of ITS and 16S metagenomic DNA. Kiwifruit pathogens associated with postharvest rot, such as Alternaria, Didymella, and Cladosporium, were significantly more prevalent on kiwis grown without nets. Additionally, different net types influenced microbial diversity, richness, and network structure. Pearl nets promoted bacterial richness and fungal diversity, while yellow nets enhanced overall diversity and resilience in both microbial communities. Grey nets resulted in evenness in fungal communities but led to less robust bacterial networks. Kiwifruit yield increased under photoselective nets compared to outside. At harvest, fruit dry matter, firmness, pH, total soluble solids, and titratable acidity were similar across treatments. However, fruits under yellow and grey nets showed significantly lower firmness compared to other treatments. Understanding these effects may contribute to optimizing fruit production and shelf-life management.

RevDate: 2025-12-30

Chen L, Tang C, Hu D, et al (2025)

Brevilin a reverses colitis of inflammatory bowel disease via modulation of TNF-α signaling and microbiome dysregulation.

Gut pathogens pii:10.1186/s13099-025-00792-3 [Epub ahead of print].

BACKGROUND: Brevilin A (Br) has shown potential in modulating inflammatory bowel disease (IBD). Our study aims to explore its mechanism of anti-inflammatory action.

METHODS: Colitis was induced in C57BL/6 mice with dextran sulfate sodium (DSS), followed by treatment with or without Br(20 mg/kg). Fecal microbiota and metabolites were profiled by metagenomic sequencing and liquid chromatography-mass spectrometry (LC-MS), respectively. Furthermore, to delineate the essential role of the gut microbiota, we employed antibiotic-treated (microbiota-depleted) mice in our investigation of Br's mechanism of action.

RESULTS: Br significantly alleviated DSS-induced colitis and modulated the gut microbiota profile. Specifically, Br enriched beneficial bacteria such as Lactobacillus, while suppressing pathogenic bacteria including Escherichia coli and Clostridium perfringens. Metabolomic analysis revealed that Br significantly altered bacterial metabolites, including 7-Oxolithocholic Acid, Kudinoside A, Veratrine, and Soyasaponin. These metabolites were linked to key pathways such as GPCR signaling, DNA damage response, aminoacyl-tRNA biosynthesis, riboflavin metabolism, and central carbon metabolism in cancer. Transcriptomic profiling indicated that Br inhibited the TNF-α signaling pathway, and this inhibition was confirmed as TNF-α overexpression reversed its anti-inflammatory effects. Furthermore, the therapeutic effects of Br were partially recapitulated in microbiota-depleted mice through fecal microbiota transplantation from Br-treated donors.

CONCLUSION: Br's ability to regulate gut microbiota and metabolites, improve gut barrier function, and eliminate inflammation by inhibiting TNF-α highlights its potential as a novel therapeutic medicine for IBD. Future research should focus on further exploring its mechanisms and clinical applications.

RevDate: 2025-12-30
CmpDate: 2025-12-30

Zhao L, Li M, Wang Y, et al (2025)

Combined effects of sound and temperature on the composition and function of bacterial and fungal communities in loess.

BMC microbiology, 25(1):803.

In Northwest China, the dominant soil type is loess, which is highly susceptible to various environmental factors. Of these, limited research has focused on the impacts of sound disturbance and temperature fluctuations on the microbial communities in loess. An orthogonal experiment was conducted by varying sound intensity (70 dB, 90 dB, 110 dB), sound duration (2 h, 4 h, 6 h), and temperature (- 5 °C, 15 °C, 35 °C). Metagenomic sequencing was then applied to investigate the effects of sound and temperature on the composition and function of bacterial and fungal communities in loess. Our results show that under the combined effects of sound and temperature, the dominant phyla and genera of bacteria and fungi have different responses and preferences to temperature and sound decibels. Alpha diversity analysis revealed that the Shannon index of the bacterial community differed significantly under the 90 dB treatment at - 5 °C and under the 110 dB treatment at 15 °C (P < 0.05). For the fungal community, both the Simpson and Shannon indices showed significant differences under the 70 dB treatment at - 5 °C and under the 110 dB treatment at 15 °C (P < 0.05). Notably, the richness of rare fungal taxa and overall species richness in the loess fungal community were significantly enhanced at 90 dB compared with the control and other treatment groups, while these indices were significantly reduced at 110 dB. In the loess microbial treatment groups subjected to the combined effects of sound and temperature, the gene abundance of CAZy family genes was lowest under high decibel (110 dB) sound stimulation. Among the six enzyme-encoding gene categories within the CAZy family, the highest number of annotated species was observed in Group A (2 h, 70 dB, - 5 °C), whereas the lowest was recorded in Group C (6 h, 110 dB, - 5 °C). Among the metabolic pathway functional genes annotated in the KEGG database, the abundance of metabolic genes in Group C (6 h, 110 dB, - 5 °C) was significantly lower than that in other treatment groups.

RevDate: 2025-12-30
CmpDate: 2025-12-30

Li J, Zhang X, Zhao X, et al (2025)

Characterising gut microbiome dysbiosis in diarrhoea calves from multiple farms in Inner Mongolia using 16S and metagenomics.

Microbiome, 13(1):259.

BACKGROUND: The pathogenesis of neonatal calf diarrhoea (NCD), a critical disease that contributes to neonatal mortality in calves, remains nebulous.

RESULTS: Inner Mongolia, a key region for cattle farming in China, was selected as a study area to provide a comprehensive overview of the epidemiology and treatment of calf diarrhoea. No significant correlation was found between the incidence of diarrhoea and sampling points or medications. The severity of diarrhoea cases was stratified into five levels based on faecal characteristics. To elucidate the pathogenesis of NCD, 16S rRNA gene and metagenomic sequencing analyses were performed across severity levels. Microbial diversity analyses revealed distinct variations in microbial communities at different severity levels. Employing binning and LEfSe methodologies, two potential bacterial pathogens were identified: Escherichia coli (bin.216), leveraging non-canonical virulence mechanisms; and Streptococcus ruminantium (bin.338), an uncharacterised diarrhoeagenic bacterium. Furthermore, the viral agent Escherichia phage VpaE1_ev108 was significantly associated with disease progression. Gene function enrichment analysis revealed a broad spectrum of antibiotic resistance genes even in farms without direct antibiotic treatment, underscoring the pervasive prevalence of drug resistance.

CONCLUSIONS: The findings of this study revealed significant gut microbial dysbiosis in calves with severe diarrhoea, through which two putative NCD-associated pathogens were identified: E. coli (bin.216) and S. ruminantium (bin.338). Marked enrichment of Bacteroides spp. and Methanobrevibacter_A sp. 900313645 was observed in healthy cohorts, suggesting their potential protective roles. Therapeutic strategies employing phage-mediated pathogen targeting combined with probiotic transplantation have demonstrated dual benefits, potentially reducing antimicrobial dependency and preserving microbial homeostasis through ecological network reconstruction. Video Abstract.

RevDate: 2025-12-30
CmpDate: 2025-12-30

Thirumaran S, George C, Kortheerakul C, et al (2025)

Fungi dominate eukaryotic microbial assemblages in hot spring biofilms of East and Southeast Asia.

Environmental microbiome, 20(1):156.

Terrestrial hot springs are globally distributed extreme environments, and these systems have long served as natural laboratories for studying microbial life under thermal stress. While much of the research to date has focused on thermophilic bacteria and archaea, there is a growing appreciation for the diversity and ecological significance of eukaryotic microorganisms in these habitats. In this study, we used metagenomic sequencing to assess inter-domain microbial diversity in biofilms from 47 circumneutral hot springs across East and Southeast Asia, with a specific focus on resolving eukaryotic taxa and their ecology. Whilst all biofilm communities were dominated by bacteria, the microbial eukaryotes represented approximately 10% of the taxonomic diversity and accounted for 1.3% of overall taxa abundance, indicating a small but significant presence. We provide the first comprehensive inter-domain checklist of over 14,500 microbial taxa in hot springs. Patterns in diversity were significantly correlated with temperature, hydrogen sulfide, and pH in hot springs. Fungi emerged as the most abundant and prevalent eukaryotic group, indicating an important role as eukaryotic saprotrophs, with Ascomycota yeasts comprising the most individually abundant taxa. Among other microbial eukaryotic phyla, the photosynthetic Chlorophyta and Bacillariophyta were most abundant. Predatory/grazing microbial eukaryotes were relatively less diverse and abundant. Network co-occurrence analysis was used to indicate extensive and specific biotic interactions between eukaryotes and bacteria in biofilms. We further employed metatranscriptomics to identify putatively active taxa, revealing that most detected eukaryotes were transcriptionally active. While fungi accounted for most transcripts, the highest RNA:DNA ratios were observed among predatory and photosynthetic taxa, suggesting elevated activity in these functional groups. Overall, our findings highlight the diversity, interactions, and activity of eukaryotes in Southeast Asian hot spring biofilms, underscoring their potential importance in shaping microbial community structure and function in extreme environments.

RevDate: 2025-12-30
CmpDate: 2025-12-30

Zhang H, Shen C, Lei W, et al (2025)

Pilot Clinical Trial of Fecal Microbiota Transplantation for Constipation in Parkinson's Disease.

Journal of microbiology and biotechnology, 35:e2509029 pii:jmb.2509.09029.

The purpose of this study was to evaluate the safety and efficacy of fecal microbiota transplantation in patients with constipation due to parkinson's disease. Gut dysbiosis has long been associated with parkinson's and recent studies have shown that FMT can restore the normal flora of the gut. Therefore, this clinical trial aimed to test the therapeutic efficacy of FMT in 5 patients aged 55 to 71 diagnosed with PD who presented with constipation. The study was conducted as an open label, prospective trial and consisted of FMT performed every 3 days via nasojejunal tube placement followed by 8 weeks of patient follow-up to evaluate response to drug therapy and to assess neurological function using UPDRS-III OFF scores, and improvement in constipation assessed with Wexner scores. Samples taken before and after FMT were collected for shotgun metagenomic sequencing to analyze the composition of the microbial communities present in patients. Untargeted non-targeted metabolomic studies were performed to investigate the impact of FMT on metabolome changes due to FMT. The results indicate an improvement in constipation and neurological functioning following FMT, and significant alteration of the gut microbiota. Significant increases in Bifidobacteria bifidus, Alistipes shahi, Anaerotruncus coli, and uncharacterized Flavonifractor were found post-treatment compared to the baseline. Many of the other strains present prior to treatment, including Acinetobacter sp. and Proteobacteria sp., had significantly decreased after the FMT. The metabolomic studies found shifts in metabolic pathways involved with unsaturated fatty acid synthesis and amino acid metabolism due to FMT. FMT may be an effective treatment option for constipation and neurological symptoms associated with PD.

RevDate: 2025-12-30
CmpDate: 2025-12-30

Li S, M Zhang (2025)

Mycobacterium chelonae infection treated with combined medication and surgical therapy: A case report.

Medicine, 104(52):e46692.

RATIONALE: The misuse of antibiotics and immunosuppressive agents has led to an increase in the incidence of infections caused by opportunistic pathogens, and even rare microbial infections such as Mycobacterium chelonae (MC) have emerged. Currently, there are very few reported cases of MC infection globally, and clinical experience in treating this disease is limited. This case report focuses on the treatment methods for infections caused by this bacterium, aiming to accumulate experience for clinicians in the diagnosis and treatment of this condition.

PATIENT CONCERNS: This study reports a case of MC infection that occurred following an invasive cosmetic injection treatment. Due to the progression of the condition, medical intervention was required. The patient underwent combined medication and surgical therapy, and the treatment outcomes were subsequently tracked.

DIAGNOSES: Histopathological examination of the skin lesion, fungal culture, and metagenomic detection of the infectious pathogen suggested MC infection.

INTERVENTIONS: The patient underwent excision of the skin lesion, along with intravenous infusion of levofloxacin hydrochloride and sodium chloride injection at a dose of 0.4 g once daily and oral administration of rifampicin capsules at a dose of 0.45 g once daily. Complete lesion resolution was achieved at 1 month postoperatively.

OUTCOMES: At the 1-month postoperative follow-up, the patient's rash had healed completely.

LESSONS: This case fills a gap in domestic and international reports of MC infection following cosmetic filler injections. By employing metagenomic next-generation sequencing, the pathogen was rapidly identified within a short timeframe, significantly reducing the diagnostic delay associated with conventional bacterial culture and identification methods. Early radical lesion excision combined with a dual-antibiotic regimen (levofloxacin-rifampin) achieved complete resolution within 1 month, establishing a replicable treatment paradigm for cutaneous MC infections. Further multicenter prospective studies are warranted to optimize surgical margins and antimicrobial treatment duration.

RevDate: 2025-12-30
CmpDate: 2025-12-30

Angelova B, Boteva S, Traykov I, et al (2025)

Bacterial Community Composition and Structure in the Littoral of Rila Mountains Glacial Lakes.

Life (Basel, Switzerland), 15(12):.

High-mountain lakes are biodiversity hotspots sensitive to increasing regional and global climate warming. However, their microbial communities remain insufficiently characterized due to their remoteness and limited accessibility. This study aimed to determine how seasonal environmental parameters shape the composition, structure and diversity of littoral bacterial communities in three glacial lakes in Rila Mountains (Bulgaria). Water samples were collected during ice-free periods in 2023 and 2024, and bacterial taxonomic composition was analysed by Next-generation sequencing. A total of 1158 bacterial OTUs were identified encompassing 18 phyla and 165 families. Actinomycetota, Pseudomonadota, and Bacteroidota were dominant at the phylum level, and Sporichthyaceae, Comamonadaceae, Chitinophagaceae and Mycobacteriaceae were most abundant among the families. Community richness and diversity peaked in June, immediately after ice melting, particularly in the highest-altitude lake (Sulzata Lake), and declined during the warm season (August), when the relative abundances of Sporichthyaceae and Mycobacteriaceae (Actinomycetota) increased. Seasonal restructuring occurred across phyla and families even in a single taxon, with water temperature and organic carbon availability identified as the main environmental drivers. The findings have improved our understanding of temperature-driven bacterial responses. They have also highlighted the vulnerability of cold-adapted taxa to regional climate warming which may contribute to more effective biodiversity conservation strategies for these unique ecosystems.

RevDate: 2025-12-30
CmpDate: 2025-12-30

Jin JX, Wang Y, Zhang GF, et al (2025)

Diversity and Functional Analysis of Gut Microbiota in the Adult of Lissorhoptrus oryzophilus (Coleoptera: Curculionidae) by Metagenome Sequencing.

Insects, 16(12):.

The gut microbiota of insects plays a fundamental role in modulating host physiology, including nutrition, development, and adaptability to environmental challenges. The rice water weevil, Lissorhoptrus oryzophilus Kuschel (Coleoptera: Curculionidae), is a major invasive pest of rice worldwide, yet the composition and functional profile of its gut microbial community remain poorly characterized. Here, we employed metagenome sequencing on the Illumina NovaSeq X Plus platform to explore the gut microbial diversity and predicted functions in adults of L. oryzophilus. Our results revealed a rich microbial community, comprising 26 phyla, 42 classes, 72 orders, 111 families, and 191 genera. The bacterial microbiota was overwhelmingly dominated by the phylum Proteobacteria (85.13% of total abundance). At the genus level, Pantoea (48.86%) was the most predominant taxon, followed by Wolbachia (14.57%) and Rickettsia (11.81%). KEGG analysis suggested that the gut microbiota is primarily associated with metabolic pathways such as membrane transport, carbohydrate and amino acid metabolism, cofactor and vitamin metabolism, energy metabolism, and signal transduction. eggNOG annotation further highlighted significant gene representation in amino acid and carbohydrate transport and metabolism, while CAZy annotation revealed glycosyl transferases (GTs) and glycoside hydrolases (GHs) as the dominant carbohydrate-active enzymes. This study provides the first comprehensive insight into the gut microbiome of L. oryzophilus adults, highlighting its potential role in the ecological success of this invasive pest. Our findings lay groundwork for future research aimed at developing novel microbial-based strategies for the sustainable management of L. oryzophilus.

RevDate: 2025-12-30
CmpDate: 2025-12-30

Zhao H, Wang H, Zhao X, et al (2025)

Bifidobacterium adolescentis Strengthens Gut Barrier in Post-Voyage Functional Constipation.

International journal of molecular sciences, 26(24):.

Prolonged periods of sailing may contribute to the development of functional constipation, which can significantly impair an individual's work efficiency. Currently, the efficacy of Bifidobacteria in treating functional constipation is gaining recognition. However, since the therapeutic effects of Bifidobacteria are strain-specific, further research is required on strains isolated from pre-voyage fecal samples. This study examines the role of gut microbiota in post-stroke constipation, aiming to identify specific microbial biomarkers for the development of targeted therapeutic strategies. B. adolescentis was identified through metagenomic analysis and subsequently isolated for validation. In the experimental group (EG), C57BL/6J mice received fecal suspension treatment following a 12-day navigation period, which was subsequently followed by a 12-day oral administration of B. adolescentis. After treatment, EG significantly improved fecal volume, intestinal motility, and goblet cells; reversed microbial ecological imbalance; reduced pathogens (E. coli and Klebsiella) by restoring arginine/bile acid metabolism, decreasing Tauro-ursodeoxycholic acid (TUDCA) content, 5-Hydroxytryptamine 4 Receptor (5-HT4R)/Slc8a1 signaling, and Ca[2+] signaling pathway; and restoring beneficial species (B. adolescentis, Pseudomonas aeruginosa). This study provides new insights into probiotics in improving human intestinal health.

RevDate: 2025-12-30

Chang Z, Chen J, Chen J, et al (2025)

Precision diagnosis of preoperative infection in urolithiasis: integrating targeted next-generation sequencing for enhanced accuracy-a multicenter cohort study.

BMC infectious diseases, 25(1):1767.

BACKGROUND: Preoperative infection in urolithiasis patients poses significant risks for surgical complications. Traditional urine culture methods have limitations in detection speed and accuracy, necessitating more precise and rapid diagnostic approaches for preoperative infection assessment.

METHODS: This multicenter cohort study evaluated urolithiasis patients with suspected preoperative infections (discovery cohort: n = 114; validation cohort: n = 64), comparing the diagnostic performance of traditional urine culture, metagenomic next-generation sequencing (mNGS), and targeted next-generation sequencing (tNGS).

RESULTS: In urolithiasis patients with suspected preoperative infections, both NGS methods demonstrated 100% sensitivity and negative predictive values (NPV) for pathogen detection, with tNGS showing superior specificity and positive predictive values (PPV) compared to mNGS. tNGS significantly reduced detection time (13 h vs. 48–72 h for traditional culture). mNGS detected 43 distinct pathogens while tNGS identified 42, both revealing previously undetected microorganisms including viruses and fungi. Among culture-positive cases, Escherichia coli (E. coli) was the predominant pathogen with higher prevalence in female patients. In polymicrobial infection patterns, tNGS identified bacterial-viral combinations as the dominant pattern (50.79%), followed by bacterial-mycoplasma/chlamydia (17.46%), and bacterial-viral-fungal (12.70%) combinations. These findings were validated in the second cohort, where bacterial-viral co-infections remained predominant (73.68%). Both NGS methods maintained 100% concordance with culture-positive results in clear and turbid urine samples, with tNGS demonstrating fewer false positives.

CONCLUSIONS: For urolithiasis patients with suspected preoperative infections, tNGS emerges as a superior diagnostic tool due to its higher specificity, faster turnaround time, and ability to detect complex pathogen patterns. While mNGS offers advantages in broad-spectrum screening, tNGS provides a more practical approach for precise preoperative infection assessment, potentially improving surgical timing decisions and patient outcomes.

CLINICAL TRIAL: Not applicable.

SUPPLEMENTARY INFORMATION: The online version contains supplementary material available at 10.1186/s12879-025-12119-3.

RevDate: 2025-12-30

Jin L, Xu Q, Miao C, et al (2025)

Dynamic multi-omics analysis reveals the correlation between aroma compounds and symbiotic microbial community during tobacco leaf aging process.

BMC plant biology, 25(1):1745.

UNLABELLED: Aging in crops like tea and tobacco involves the production of secondary metabolites, with symbiotic microbes playing a key role. However, their dynamic changes and correlation with metabolites during aging remain poorly understood. This study investigates changes in microbial communities, aroma compounds, and protein expression during tobacco leaf aging using artificial accelerated aging techniques, which combine GC-MS, metagenomics, and metaproteomics methods. We identified 62 aroma compounds with distinct change patterns and observed significant changes in the structure of symbiotic bacteria. Type one, represented by Wolbachia_endosymbiont_of_Diaphorina_citri, increased in abundance from the fourth month, correlating with compounds like 2-Furaldehyde. Type two, represented by Sphingomonas_sp_LK11, showed a bimodal abundance pattern, correlating with compounds like Tabanone. Metaproteomics revealed that protein functions were initially limited to cytoskeleton organization but diversified from the fourth month. Fungi also displayed two distinct clustering patterns, Rhizopus and Mortierella elongata were abundant early on, while Colletotrichum asianum and Trichophyton violaceum appeared later. Rhizopus and other fungi exhibited a significant positive correlation with 24 aroma compounds, including 5-Methylfuran-2(5 H)-one. Linderina pennispora and other fungi showed a significant positive correlation with 28 aroma compounds, including 2-Furaldehyde. The dynamic changes in microbial community structure during aging are closely related to the generation of aroma compounds. Overall, temporal shifts in microbial communities were closely linked to aroma formation. One set of microorganisms, such as Wolbachia_endosymbiont_of_Diaphorina_citri and Linderina pennispora, is positively correlated with 2-Furaldehyde, Isophorone, and 2-Methylbenzofuran. Another set, including Sphingomonas_sp_LK11 and Rhizopus, exhibits a positive correlation with 5-Methylfuran-2(5 H)-one and 1,2-Cyclohexanedione. These findings provide new insights into the biological mechanisms of tobacco leaf aging, and offer new research directions for the development and innovation of future tobacco products.

SUPPLEMENTARY INFORMATION: The online version contains supplementary material available at 10.1186/s12870-025-07765-3.

RevDate: 2025-12-30
CmpDate: 2025-12-30

Xu LX, Wang F, Yao Y, et al (2025)

Key role of microbial necromass and iron minerals in retaining micronutrients and facilitating biological nitrogen fixation in paddy soils.

Fundamental research, 5(6):2612-2621.

Paddy fields, as the largest anthropogenic wetlands on Earth, face a high risk of micronutrient loss through surface runoff and leaching due to their frequent irrigation-drainage cycles, as well as removal with crop harvest. While micronutrient's losses largely impede biological nitrogen fixation (BNF) in soils, agricultural practices and mechanisms that retain micronutrients and thus increase BNF in paddy soils remain underexplored. Here we showed that the long-term (40 years) application of fertilizers increased the contents of microbial necromass in paddy soils by 20%-43% compared to the unfertilized control. In particular, long-term organic fertilizations increased poorly crystalline minerals through intensifying mineral weathering, which further contributed to the increased stable carbon burials in paddy soils. Synchrotron radiation based spectromicroscopy analysis provided direct evidence demonstrating a differential control of hydroxyl functional groups from mineral surfaces on C functional groups at the submicron scale in paddy soils. Notably, microbial necromass and short-range ordered minerals had a strong correlation with the content and bioavailability of micronutrients in paddy soils, indicating their key roles in the retention of micronutrients. Metagenomic sequencing analysis further indicated that the content and bioavailability of micronutrients were strongly correlated with the abundance of the key N-fixing genera (i.e., Azospirillum and Bradyrhizobium). Unexpectedly, structural equation modeling (SEM) identified that microbial necromass exerted the strongest control on N-fixing genera, highlighting an underappreciated role of microbial necromass as a reservoir of micronutrients. Based on micronutrient's bioavailability and metagenomic sequencing, we conclude that micronutrients are the key factor for BNF in paddy soils, offering significant implications for managing BNF in paddy soils.

RevDate: 2025-12-30
CmpDate: 2025-12-30

Sokolova EA, Smirnova NV, Fedorets VA, et al (2025)

Microbial Consortium Application Under Temperature Stress: Effects on the Rhizosphere Microbiome and Plant Growth.

International journal of molecular sciences, 26(24): pii:ijms262411814.

The aim of the present study was to investigate the effect of a synthetic microbial consortium (SMC) containing five functionally different bacterial strains (Rahnella aquatilis, Rothia endophytica, Stenotrophomonas indicatrix, Burkholderia contaminans, Lelliotia amnigena) on the growth and development of three agricultural crops (wheat, buckwheat, and rapeseed) on two soil types (chernozem and gray forest soil) under field conditions. The experiment was conducted from June to September 2024 under extreme field conditions, with temperatures reaching 43.8 °C. This study evaluates SMC efficacy under severe abiotic stress, reflecting increasingly common climate extremes. Metagenomic data analysis showed that the introduced strains did not establish stable populations in the soil, possibly due to heat-induced bacterial mortality, though other factors including competition with indigenous microflora and lack of protective formulations may have also contributed. No statistically significant effects on plant morphometric parameters were observed. The extreme temperature and water stress conditions appear to have been the dominant limiting factors, overriding any potential benefits from microbial inoculation, as evidenced by the lack of response to mineral fertilizer application as well. Crop-specific effects were revealed: when cultivating rapeseed on chernozem, a significant increase in available phosphorus content was noted (from 278 ± 45 to 638 ± 92 mg/kg with SMC application, p < 0.001).

RevDate: 2025-12-30
CmpDate: 2025-12-30

Neumann GB, Korkuć P, Rahmatalla SA, et al (2025)

Surviving the Heat: Genetic Diversity and Adaptation in Sudanese Butana Cattle.

Genes, 16(12): pii:genes16121429.

BACKGROUND: Butana are native Sudanese Bos indicus cattle that are well adapted to arid environments and valued for their relatively high milk performance and resilience under harsh conditions. Despite their adaptive advantages, Butana cattle face the risk of genetic erosion due to low production performance and the absence of structured breeding programs underscoring the urgent need to conserve their unique genetic potential for climate-resilient livestock development.

METHODS: In this study, we analyzed whole-genome sequencing data from 40 Butana cattle to assess their genetic diversity, population structure, signatures of selection, and potential pathogen load.

RESULTS: Butana cattle exhibited high nucleotide diversity and low levels of inbreeding, indicating a stable gene pool shaped by natural selection rather than by intensive breeding. Signatures of selection and functional variant analysis revealed candidate genes involved in heat stress adaptation (COL6A5, HSPA1L, TUBA8, XPOT), metabolic processes (G6PD, FAM3A, SLC10A3), and immune regulation (IKBKG, IRAK3, IL18RAP). Enrichment analyses and RoH island mapping consistently highlighted immune and thermoregulatory pathways as key selection targets, distinguishing Butana from both the geographically neighbored Kenana cattle and the specialized dairy cattle breed Holstein. Furthermore, metagenomic screening of unmapped reads detected the tick-borne parasite Theileria annulata and the opportunistic pathogen Burkholderia cenocepacia in all animals, underscoring the importance of integrating pathogen surveillance into genomic studies.

CONCLUSIONS: Taken together, our findings highlight the distinct adaptive genomic profile of Butana cattle and reinforce their value in breeding programs aimed at improving climate resilience and disease resistance in livestock through the utilization of local breeds.

RevDate: 2025-12-30
CmpDate: 2025-12-30

Kahraman Ilıkkan Ö, Bağdat EŞ, Yılmaz R, et al (2025)

Bridging Tradition and Innovation: A Systematic Review and Bibliometric Analysis of Turkish Fermented Foods.

Foods (Basel, Switzerland), 14(24): pii:foods14244324.

BACKGROUND: Traditional fermented foods from Türkiye are integral components of the nation's culinary heritage, reflecting a remarkable diversity shaped by local practices and ecosystems. These products embody region-specific microbial communities, often conceptualized as a "microbial terroir," which influence their sensory qualities, nutritional value, and health-promoting properties.

METHODS: This study followed the PRISMA 2020 guidelines and included a systematic review and bibliometric analysis. A structured search was conducted in the Web of Science Core Collection (WoSCC) on 15 January 2025 using predefined keywords related to Turkish fermented foods and fermentation processes. Records were screened based on language (English or Turkish) and document type (articles, reviews, book chapters, and early access). A total of 1464 studies met the eligibility criteria, reflecting a 2.81% annual growth rate and a 12.7% international co-authorship rate. Bibliometric analysis was performed using the bibliometrix R package (RStudio 2024.12.1) and the biblioshiny interface.

RESULTS: The analysis revealed that the diversity of microbial consortia in Turkish fermented foods contributes to their distinctive characteristics, including enhanced nutritional profiles, probiotic potential, and food safety attributes. Emerging studies employing omics technologies-such as next-generation sequencing, metagenomics, and metabolomics-have expanded our understanding of fermentation ecosystems. Additionally, the growing integration of artificial intelligence supports predictive modeling and process optimization for advanced quality control.

CONCLUSION: This synthesis highlights the significant technological, nutritional, and cultural value of Türkiye's traditional fermented foods. Future directions should include omics-based translational research, indigenous starter culture development, and strengthened international collaborations to support sustainable and competitive functional food innovation.

RevDate: 2025-12-30
CmpDate: 2025-12-30

Ospałek W, Wlazło Ł, Tajchman K, et al (2025)

Cultivated Gut Microbiota of Roe Deer and Red Deer in Central Poland Forest.

Animals : an open access journal from MDPI, 15(24): pii:ani15243656.

The aim of this study was to compare, using culture methods, the microflora of the small and large intestines, distinguishing pathogenic bacteria, in free-living ruminants: roe deer (Capreolus capreolus) and red deer (Cervus elaphus). Intestinal samples from six individuals of each species were collected immediately after hunting under aseptic conditions. Aerobic and facultatively anaerobic bacteria, including Lactobacillus spp., Escherichia coli, Listeria spp., and Clostridium perfringens, were quantified using standard culture methods. Statistical analysis (ANOVA) revealed no significant differences (p > 0.05) between species in any of the microbial groups analyzed, although higher mean abundances were observed in red deer, particularly in the large intestine. The results indicate that interspecific variation in cultured microbiota may reflect individual and environmental factors rather than consistent taxonomic differences. Due to the high inter-individual variability and limited sample size, this study should be considered preliminary. The results demonstrate the predominance of viable aerobic and facultative anaerobic bacterial groups in culture-based analysis and provide reference data for future metagenomic studies. This study fills an important knowledge gap, as culture-dependent studies of the gut microbiota of wild cervids are still rare due to the logistical and ethical constraints associated with sampling wild animals.

RevDate: 2025-12-30
CmpDate: 2025-12-30

Woś K, Pachciński K, Wacko M, et al (2025)

The Role of Swine Gut Microbiota and Its Metabolites in Maintaining Intestinal Barrier Integrity and Mitigating Stress via the Gut-Brain Axis.

Animals : an open access journal from MDPI, 15(24): pii:ani15243653.

The gut-brain axis is a complex communication network linking the gastrointestinal tract and the central nervous system, in which the gut microbiota plays a pivotal role in regulating intestinal homeostasis, immune responses and neuroendocrine functions. This review summarizes current knowledge on the role of the porcine gut microbiota in the functioning of the gut-brain axis and examines nutritional strategies aimed at its modulation. Key production-related stressors, such as weaning, transport and handling, disrupt microbiota composition, increase intestinal permeability and activate the hypothalamic-pituitary-adrenal (HPA) axis, leading to heightened stress responses, impaired immunity and behavioral disturbances. Evidence indicates that supplementation with probiotics, prebiotics, or postbiotics stabilizes the gut microbiota, enhances the production of bioactive metabolites, supports intestinal barrier integrity and alleviates oxidative stress. Such interventions improve adaptation to environmental stress, animal welfare and performance, while potentially reducing the need for antibiotics. Maintaining a balanced gut microbiota is therefore essential for the proper functioning of the neuroendocrine and immune systems in pigs. An integrated approach utilizing omics technologies (metagenomics, metabolomics, proteomics) may further elucidate microbiota-brain interactions and support the development of sustainable and ethical swine production strategies.

RevDate: 2025-12-30
CmpDate: 2025-12-30

Sakarnyte L, Spinkyte R, Merkeviciene L, et al (2025)

Next-Generation Sequencing Insights into the Oral Microbiome and Antibiotic Resistance Genes in Grey Wolves (Canis lupus).

Animals : an open access journal from MDPI, 15(24): pii:ani15243639.

The oral microbiome of apex predators such as grey wolves (Canis lupus) is colonised by complex microbial communities and plays a crucial role in the health of wild mammals, but remains poorly understood. In this study, a single pooled sample mixed from oral samples of 17 wolves (Canis lupus) hunted in Lithuania was investigated for the determination of a variety of oral microbiota, providing the first metagenomic insight into wolf oral microbiomes in Baltic countries. The aim of this study was to identify the zoonotic and antimicrobial resistance potential of the oral microbiota. The results revealed diverse microbiomes associated with periodontal health as well as microbiomes related to the environment. Unique microbial taxa potentially reflect the nutritional and ecological interactions of carnivores. Shotgun metagenomic sequencing yielded a total of 18,726,406 raw reads, and following quality trimming and filtering, 86.01% of these (16,106,613) were retained. Among the total reads, 45.15% (8,455,255) were identified as host-derived and were removed. The most common oral bacterial genera were Pseudomonas (50%) and Psychrobacter (22.6%). Metagenomic reads for zoonotic pathogens, including Salmonella, Mycobacterium spp., Yersinia, Coxiella burnetii, Corynebacterium pseudotuberculosis, and others, were also detected, suggesting that grey wolves are potential natural reservoirs of zoonotic infections. Genes encoding antimicrobial resistance to many classes of antibiotics were also detected. This research contributes to understanding wolf dietary habits, oral health, the carriage and possible risk of transmitting AMR, and social interactions.

RevDate: 2025-12-30
CmpDate: 2025-12-30

Moreira G, Rodrigues S, Gomes-Gonçalves S, et al (2025)

Highly Virulent Newcastle Disease Virus in Eurasian Collared Doves in the North of Portugal.

Animals : an open access journal from MDPI, 15(24): pii:ani15243563.

Newcastle disease (ND), caused by avian orthoavulavirus 1 (AOAV-1), poses a global threat to poultry and wild birds. In early 2025, an outbreak of pigeon paramyxovirus type 1 (PPMV-1, genotype VI AOAV-1) was detected in a wildlife rehabilitation centre in northern Portugal, affecting Streptopelia decaocto, Streptopelia risoria, and Columba livia. Birds showed acute neurological signs and died rapidly. Necropsy revealed brain and pulmonary congestion, splenomegaly, and cloacal lesions, while histopathology demonstrated hepatocellular necrosis, hemorrhage, and eosinophilic intracytoplasmic inclusions in hepatocytes and renal tubular cells. Matrix (M) gene PCR using standard primers was negative, but metagenomic sequencing identified genotype VI as being closely related to strains from Iran and Cyprus. Partial fusion (F) gene analysis revealed the velogenic RRQKRF motif. These findings confirm the circulation of highly virulent PPMV-1 in Portugal, highlight that standard, recommended primers may fail to detect some genetically diverse strains, and emphasize the role of Columbidae as reservoirs with potential transmission to domestic poultry.

RevDate: 2025-12-30
CmpDate: 2025-12-30

Novakovic J, Milosavljevic I, Stepanova M, et al (2025)

Safe Meat, Smart Science: Biotechnology's Role in Antibiotic Residue Removal.

Antibiotics (Basel, Switzerland), 14(12): pii:antibiotics14121264.

The widespread use of antibiotics in livestock farming has led to the persistent issue of antibiotic residues in meat products, raising significant concerns for food safety and public health. These residues can contribute to the emergence and spread of antimicrobial resistance (AMR), a growing global health threat recognized by the World Health Organization. While some regulatory bodies have imposed restrictions on non-therapeutic antibiotic use in animal agriculture, inconsistent global policies continue to hinder unified efforts to reduce AMR risks. This review explores the role of biotechnology in addressing this challenge by offering innovative tools for the detection, degradation, and removal of antibiotic residues from meat. Biotechnological approaches include the use of biosensors, high-throughput screening, enzymatic degradation, microbial bioremediation, genetically engineered bacteria, phage therapy, and phytoremediation. In addition, enabling technologies such as genomics, metagenomics, bioinformatics, and computational modeling support the rational design of targeted interventions. We further examine the integration of these biotechnological strategies within the broader "One Health" framework, which emphasizes the interconnectedness of human, animal, and environmental health. Case studies and recent applications demonstrate the potential of these methods to ensure safer meat production, reduce public health risks, and enhance consumer trust. By focusing on scalable, science-driven solutions, biotechnology offers a promising path toward mitigating antibiotic residues in the food supply and combating the long-term threat of AMR.

RevDate: 2025-12-30
CmpDate: 2025-12-30

Saleem N, Kumar N, El-Omar E, et al (2025)

Harnessing Machine Learning Approaches for the Identification, Characterization, and Optimization of Novel Antimicrobial Peptides.

Antibiotics (Basel, Switzerland), 14(12): pii:antibiotics14121263.

Antimicrobial resistance (AMR) has become a major health crisis worldwide, and it is expected to surpass cancer as one of the leading causes of death by 2050. Conventional antibiotics are struggling to keep pace with the rapidly evolving resistance trends, underscoring the urgent need for novel antimicrobial therapeutic strategies. Antimicrobial peptides (AMPs) function through diverse, often membrane-disrupting mechanisms that can address the latest challenges to resistance. However, the identification, prediction, and optimization of novel AMPs can be impeded by several issues, including extensive sequence spaces, context-dependent activity, and the higher costs associated with wet laboratory screenings. Recent developments in artificial intelligence (AI) have enabled large-scale mining of genomes, metagenomes, and quantitative species-resolved activity prediction, i.e., MIC, and de novo AMPs designed with integrated stability and toxicity filters. The current review has synthesized and highlighted progress across different discriminative models, such as classical machine learning and deep learning models and transformer embeddings, alongside graphs and geometric encoders, structure-guided and multi-modal hybrid learning approaches, closed-loop generative methods, and large language models (LLMs) predicted frameworks. This review compares models' benchmark performances, highlighting AI-predicted novel hybrid approaches for designing AMPs, validated by in vitro and in vivo methods against clinical and resistant pathogens to increase overall experimental hit rates. Based on observations, multimodal paradigm strategies are proposed, focusing on identification, prediction, and characterization, followed by design frameworks, linking active-learning lab cycles, mechanistic interpretability, curated data resources, and uncertainty estimation. Therefore, for reproducible benchmarks and interoperable data, collaborative computational and wet lab experimental validations must be required to accelerate AI-driven novel AMP discovery to combat multidrug-resistant Gram-negative pathogens.

RevDate: 2025-12-30
CmpDate: 2025-12-30

Papamentzelopoulou M, Vrioni G, V Pitiriga (2025)

Comparative Evaluation of Sequencing Technologies for Detecting Antimicrobial Resistance in Bloodstream Infections.

Antibiotics (Basel, Switzerland), 14(12): pii:antibiotics14121257.

Bloodstream infections (BSIs) pose a significant global health challenge, particularly due to the increasing prevalence of antimicrobial resistance (AMR). Timely and accurate identification of pathogens and resistance determinants is critical for guiding appropriate therapy and improving patient outcomes. Traditional culture-based diagnostics are limited by prolonged turnaround times and reduced sensitivity, especially in culture-negative or polymicrobial infections. This review systematically examined current and emerging sequencing technologies for AMR detection in BSIs, including whole-genome sequencing (WGS), targeted next-generation sequencing (tNGS), metagenomic next-generation sequencing (mNGS), and long-read sequencing platforms (Oxford Nanopore, PacBio). We compared their clinical performance using key metrics such as diagnostic sensitivity, turnaround time, and cost, highlighting contexts in which each technology is most effective. For example, tNGS can achieve the rapid detection of known resistance genes within 8-24 h, while WGS provides comprehensive genome-wide resistance profiling over 24-48 h. mNGS offers broader detection, including rare or unexpected pathogens, although at higher cost and longer processing times. Our analysis identifies specific strengths and limitations of each approach, supporting the use of context-specific strategies, such as combining rapid targeted sequencing for common pathogens with broader metagenomic approaches for complex cases, to improve diagnostic yield and guide antimicrobial therapy. Quantitative comparisons indicate that sequencing technologies can complement conventional methods, particularly in cases where culture-based approaches fail. In conclusion, sequencing-based diagnostics offer measurable improvements in sensitivity and speed over traditional methods for AMR detection in BSIs. Future work should focus on optimizing workflows, integrating sequencing data into clinical decision-making, and validating approaches in prospective studies.

RevDate: 2025-12-30
CmpDate: 2025-12-30

Xin R, Lin H, Li Z, et al (2025)

Plasmid-Mediated Spread of Antibiotic Resistance by Arsenic and Microplastics During Vermicomposting.

Antibiotics (Basel, Switzerland), 14(12): pii:antibiotics14121230.

Background: The efficiency of vermicomposting in reducing antibiotic resistance genes (ARGs) in dairy manure may be compromised by co-pollutants like arsenic (As) and microplastics. Specifically, plasmids serving as carriers and vectors of ARGs were largely distributed in this process. However, the impact of As and microplastics on plasmids carrying ARGs during vermicomposting is largely unknown. Methods: This study utilized a controlled experimental design and applied plasmid metagenomics to investigate the individual and combined effects of As and polyethylene terephthalate (PET) microplastics on plasmid-mediated ARG dynamics during vermicomposting. Results: We found that vermicomposting alone mainly enriched non-mobilizable plasmids, while PET microplastics selectively promoted conjugative and mobilizable plasmids, whereas As significantly increased all plasmid types. Moreover, both PET or As alone and combined exposure (PET and As) increased total ARG abundance, with their combination inducing synergistic ARG enrichment despite unchanged total plasmid abundance. Furthermore, co-occurrence network analysis combined with ARGs/plasmid ratio assessments demonstrated that As influences ARGs through co-selective pressure by enriching ARGs co-localized with As resistance genes (e.g., the ars operon) on plasmids while simultaneously promoting horizontal gene transfer (HGT) via activation of oxidative stress and SOS response pathways. In contrast, PET primarily facilitates ARG dissemination through a "metabolism-resistance" coupling strategy by enriching colonizing bacteria with PET-degrading capacity. Their co-exposure formed As-enrichment hotspots on PET microplastic surfaces, functioning as a "super-mixer" that selectively screened for superbugs carrying potent resistance mechanisms (e.g., blaOXA-50 and mdtB/mdtE). Conclusions: This study provides the first plasmidome-level evidence of synergistic ARG propagation by As and PET microplastics during vermicomposting, highlighting mobile genetic elements' critical role in co-pollutant risk assessments.

RevDate: 2025-12-30
CmpDate: 2025-12-30

Trachtmann NV, Toshchakov SV, Izotova AO, et al (2025)

Cloning and Characterization of the Novel Endoglucanase Identified in Deep Subsurface Thermal Well of Biragzang (North Ossetia) by Metagenomic Analysis.

Biomolecules, 15(12): pii:biom15121710.

The phylum Armatimonadota represents one of the least characterized bacterial lineages, with only three formally described species despite its widespread distribution in various environments. Deep subsurface thermal environments harbor significant microbial diversity and represent promising sources for novel enzyme discovery through metagenomic approaches. This study reports the identification, cloning, and biochemical characterization of Cel7465, a novel endoglucanase from an uncultured GBS-DC family within the order Fimbriimonadales. The enzyme was identified through metagenomic analysis of microbial mats from the Biragzang deep thermal well (North Ossetia, Russia, 48 °C) and successfully expressed in cells of Escherichia coli strain ArcticExpress (DE3). Phylogenetic analysis assigned Cel7465 to glycoside hydrolase family 5 subfamily 36. The purified recombinant enzyme exhibited optimal activity at 55 °C and pH 8.0, with high specific activity of 4347 U/mg. The enzyme demonstrated broad pH tolerance (50% activity retained from pH 4.0 to 10.0) and notable thermal stability, retaining 60% activity after one hour at 80 °C and 20% after four hours. Remarkably, the presence of Mn[2+] ions enhanced enzyme activity more than 7-fold, while Mg[2+] and Ca[2+] ions provided moderate activation. Cel7465 represents the first biochemically characterized glycoside hydrolase from the order Fimbriimonadales, expanding our understanding of enzymatic capabilities within the understudied phylum Armatimonadota and demonstrating the continued potential of extreme environments as sources of novel industrial biocatalysts.

RevDate: 2025-12-30
CmpDate: 2025-12-30

Li T, Fang L, Chen X, et al (2025)

The Premature Infants' Gut Microbiota Assembly and Neurodevelopment (PIGMAN) Cohort Study: Protocol for a Prospective, Longitudinal Cohort Study.

Children (Basel, Switzerland), 12(12): pii:children12121644.

BACKGROUND: Early-life gut microbiota colonization plays a significant role in the neurodevelopment of infants and young children. However, the causal relationship between early-life gut microbiota colonization and neurodevelopment in preterm infants has not yet been conclusively established. Our research will initiate the PIGMAN (Premature Infants Gut Microbiota Assembly and Neurodevelopment) cohort study to systematically examine the dynamic interplay between gut microbiota developmental trajectories and neurodevelopmental processes in preterm infants.

METHODS: This study will employ a longitudinal cohort design and utilize data from the PIGMAN cohort, examining the interplay between gut microbiota metabolism and neurodevelopmental outcomes. The study design incorporates longitudinal stool sample collection, which will be analyzed through 16S rRNA gene sequencing and metagenomic shotgun sequencing, enabling comprehensive characterization of microbial community dynamics and functional metabolic pathways.

ANTICIPATED RESULTS: Advanced analytical approaches incorporating causal inference methodologies will be implemented to identify significant microbial and metabolic biomarkers associated with neurodevelopmental outcomes in preterm neonates, and to establish causal pathways between these biomarkers and neurodevelopment. These analytical advancements will facilitate the construction of predictive models that utilize temporal microbial signatures and metabolite trajectories as prognostic indicators for neurodevelopmental outcomes. Causal inference method evaluations will further reveal that specific gut-derived metabolites, particularly those involved in cholesterol metabolism and neural signaling pathways-such as bile acids and GABA (gamma-aminobutyric acid)-exhibit superior predictive capacity for cognitive development trajectories. Anticipated Conclusions: The findings will collectively suggest that longitudinal metabolic profiling of the gut ecosystem, when combined with causal network analysis, provides a novel paradigm for developing clinically actionable predictive models of neurodevelopment in vulnerable preterm populations.

RevDate: 2025-12-30
CmpDate: 2025-12-30

Vila-Nistal M, Martinez-Hernandez F, Lluesma-Gomez M, et al (2025)

Limited consensus of marine viral diversity observed across techniques.

Environmental microbiome, 20(1):157.

BACKGROUND: Viruses are fundamental to many aspects of life influencing ecosystem functions. The `number of lenses´ we use for exploring the viral diversity has expanded, yet each has limitations that constrain our view of the uncultured virosphere. It is fundamental to evaluate the different viromic approaches and sequencing methods on their ability to recover the extant viral diversity and microdiversity present in a sample. The differences in genome recovery between technologies have downstream impacts on subsequent estimates of viral diversity and function within a sample that can limit our comprehension of natural viral assemblages and their interactions with their microbial hosts.

RESULTS: Here, using the same surface seawater sample, we compare short- and long-read viromics (i.e., Illumina, PacBio-HiFi and MinION sequencing) along with high-throughput single-virus genomics (sequencing of 700 uncultured single-viruses) to explore the consensus between approaches to uncover the extant viral diversity (sequencing effort ≈1.6 Tbp). Overall, ≈42,000 viral contigs (> 10 kb) were obtained, resulting in ≈12,500 and ≈23,400 viral OTUs at the genus and species levels, respectively, infecting mostly Flavobacteriaceae and Pelagibacteracea. At the viral family level, single-virus genomics (SVG) recovered viruses with a more distinct taxonomic profile compared to other methods. At lower taxonomic resolution, only < 1% of all species and genera, including some of the most abundant viruses, were captured by all methods; reaching a value of ≈2% when only viromics excluding SVG were considered. The highest pairwise diversity consensus was observed between PacBio-HiFi and Illumina, with approximately ≈11% of PacBio-HiFi species-level vOTUs also detected by Illumina. To understand how different methods resolve the co-occurring genomic microdiversity within species, we used one of the most abundant and microdiverse viruses -the uncultured pelagiphage vSAG 37-F6, proposed to be classified as Pelagimarinivirus ubique- originally discovered by single-virus genomics, as a reference. None of the methods alone were able to assemble the complete genome, which was only achieved by combining all datasets. Similarly, none of the viral clusters at the strain level were recovered by all methods.

CONCLUSIONS: Our study suggests that the inherent bias of each method still represents a challenge for the recovery of marine viral diversity and potentially for other environmental viral samples. Nevertheless, regarding standard viromic techniques, PacBio HiFi in combination with Illumina seem to perform the best in absolute recovery of viral species and genera.

RevDate: 2025-12-30

Jin S, Cenier A, Wetzel D, et al (2025)

Microbial collagenase activity is linked to oral-gut translocation in advanced chronic liver disease.

Nature microbiology [Epub ahead of print].

Microbiome perturbations are associated with advanced chronic liver disease (ACLD), but how microorganisms contribute to disease mechanisms is unclear. Here we analysed metagenomes of paired saliva and faecal samples from an ACLD cohort of 86 individuals, plus 2 control groups of 52 healthy individuals and 14 patients with sepsis. We identified highly similar oral and gut bacterial strains, including Veillonella and Streptococcus spp., which increased in absolute abundance in the gut of patients with ACLD compared with controls. These microbial translocators uniquely share a prtC gene encoding a collagenase-like proteinase, and its faecal abundance was a robust ACLD biomarker (area under precision-recall curve = 0.91). A mouse model of hepatic fibrosis inoculated with Veillonella and Streptococcus prtC-encoding patient isolates showed exacerbation of gut barrier impairment and hepatic fibrosis. Furthermore, faecal collagenase activity was increased in patients with ACLD and experimentally confirmed for the prtC gene of translocating Veillonella parvula. These findings establish mechanistic links between oral-gut translocation and ACLD pathobiology.

RevDate: 2025-12-29

Moradi J, Berggreen E, Gerdts E, et al (2025)

Taxonomic and functional signatures of smoking and periodontitis severity in the subgingival microbiome of older adults.

npj aging pii:10.1038/s41514-025-00319-9 [Epub ahead of print].

Periodontitis and smoking are major contributors to oral and systemic health deterioration in aging adults. This study investigated the combined effects of smoking status and periodontitis severity on the subgingival microbiome in 1107 individuals aged 69-72 using shotgun metagenomic sequencing. Smoking was linked to reduced microbial diversity, enrichment of periodontal pathogens, and depletion of health-associated commensals, while increasing periodontitis severity was associated with broader dysbiotic shifts, including enrichment of canonical pathogens. The presence of overlapping taxa suggests shared dysbiotic pathways that may accelerate disease progression in older adults. Notably, the combination of smoking and severe periodontitis was characterized by enrichment of key pathogens, such as Tannerella forsythia, Fusobacterium nucleatum, Actinomyces israelii, and Mogibacterium timidum. Although former smokers showed fewer opportunistic pathogens than current smokers, their microbiomes remained altered compared to never smokers, suggesting persistent differences potentially related to past smoking. Functional profiling revealed largely additive effects of smoking and periodontitis, with enrichment of lipopolysaccharide biosynthesis, proteolysis, and sulfur metabolism, alongside depletion of commensal biosynthetic functions. Overall, the findings highlight the persistent and additive impacts of smoking and periodontitis on the subgingival microbiome, underscoring the importance of addressing both exposures jointly in long-term oral health strategies for older adults.

RevDate: 2025-12-29
CmpDate: 2025-12-29

Su Q, Du Y, Du D, et al (2026)

New insights into the anaerobic digestion of high carbon wastewater with ciprofloxacin: Methane production and ARGs inhibition.

Journal of environmental sciences (China), 161:612-621.

Ciprofloxacin (CIP), as a quinolone antibiotic, has broad-spectrum antibacterial properties and can affect methanogenic performance in anaerobic digestion (AD). While previous studies focused on synthetic wastewater, the fate of CIP in real distillery wastewater (RDW) and its impact on microbial adaptation mechanisms remain unclear (such as biotransformation pathways, population dynamics, and the enzymes involved) in RDW is largely unclear. In this study, we investigated AD performance, metabolic pathways, and antibiotic resistance gene (ARG) dynamics using real wastewater spiked with CIP (0.3-2 mg/L). Results indicate that 0.5 mg/L CIP (631.83 mL CH4/g·VS) enhanced the methane yield by 6.67 % (592.34 mL CH4/g·VS in control), correlating with upregulated enzyme in glycosis, TCA cycle, and methanogenesis (F420 increased). With full use of short-chain acids, transient volatile fatty acid (VFA) inhibition (≤ 50 mg/L on Day 3) was overcome by Day 10. Metagenomics revealed CIP promoted the production of stress proteins (e.g., cysteine synthase activity doubled). Furthermore, CIP (0.3-1 mg/L) suppressed mobile genetic elements (MGEs) encoding horizontal gene transfer, including isfinder (15.15 %) and integrases (6.25 %), while ARG and virulence factor abundances remained unchanged versus control. This study firstly shows that low-dose CIP in RDW increases methanogenesis via metabolic adaptation without exacerbating ARG risks. MGE suppression implies that CIP may lessen the possibility of ARG diffusion in AD systems. These results offer vital information for improving AD performance in the treatment of wastewater contaminated by antibiotics and developing methods to strike a balance between antibiotic removal and ARG control.

RevDate: 2025-12-29
CmpDate: 2025-12-29

Kang S, Lee JY, KS Cho (2026)

Bacterial and fungal metagenomes associated with atmospheric particulates in Republic of Korea: Comparison of PM2.5 and TSP larger than PM2.5.

Journal of environmental sciences (China), 161:400-410.

Particulate matter (PM) significantly contributes to air pollution, potentially causing health issues, with PM-associated microorganisms implicated in some cases. While studies have explored microbial concentration and structure in PM based on particle size, comprehensive analysis of microbial functional traits and environmental influences is limited. This study evaluated microbial concentrations and diversity in PM with a diameter of 2.5 µm or lower (PM2.5) and total suspended particles (TSP) greater than PM2.5 (PM>2.5) samples relative to air temperature and other factors. DNA extracted from PM2.5 and PM>2.5 filters was sequenced to characterize bacterial and fungal community structures and functional genes. Results showed that microbial concentrations and diversity were greater in PM>2.5, with similar dominant species across PM sizes. Higher air temperatures correlated with increased microbial concentrations and diversity in PM>2.5, attributed to enhanced microbial growth. An Asian dust event from the Mongolian desert disrupted the PM microbiome. Despite consistent species dominance, gene function analysis revealed abundant drug resistance pathways in bacterial communities of both particle types, while pathotroph prevalence was higher in PM2.5 fungal communities. These findings indicate that PM2.5 microbial community analysis suffices for understanding PM ecosystems, offering valuable insights for air quality management and microbial pollution control, especially concerning potential pathogens.

RevDate: 2025-12-29
CmpDate: 2025-12-29

Qian J, Bai S, Wu L, et al (2026)

Energy recovery from corn straw-based biochar@MIL-88A(Fe)-mediated anaerobic digestion of waste activated sludge under norfloxacin: Metabolism and antibiotic resistance gene fates.

Journal of environmental sciences (China), 161:350-359.

Norfloxacin (NOR), a commonly detected antibiotic in waste activated sludge (WAS), remains understudied in anaerobic digestion (AD). This study investigated the effect of NOR on WAS AD, with corn straw-based biochar modified with MIL-88A(Fe) (BM) added to enhance energy recovery during digestion. Accumulated methane production was inhibited by 41.86 % in the BM-mediated digestion system under 1 mg/L NOR. Moreover, NOR induced the build-up of volatile fatty acids (VFAs), hindering methanogenic pathways subsequently. Microbial community structure was altered, with an enrichment of bacteria responsible for NOR degradation and a 13.20 % reduction in the abundance of hydrogenotrophic methanogens under antibiotic stress. Methanogenesis was inhibited with the expression of related genes and enzymes suppressed. The high enzymatic activities of cytochrome P-450 (CYP450) and acetate kinase contributed to the high NOR biodegradation efficiency (88.79 %). Twelve typical antibiotic resistant genes (ARGs) types, including multidrug, aminoglycoside, macrolides (MLs), etc., were examined in the AD system. The total abundance of ARGs type and subtype increased under NOR addition, implying ARGs removal was inhibited by NOR stress. Resistance to NOR exposure was primarily associated with antibiotic efflux and alterations in antibiotic target. Horizontal gene transfer (HGT) and vertical gene transfer (VGT) were the mechanistic routes for ARG evolution, with HGT inhibited and VGT promoted following NOR addition. The dominant genus Acinetobacter was the potential host for nearly all ARGs. This study advanced understanding of the impact of NOR on WAS digestion with BM mediation, providing new insights for optimizing WAS digestion.

RevDate: 2025-12-29
CmpDate: 2025-12-29

Cui M, Chen S, Zhang Z, et al (2026)

Nanoplastics and triclosan co-exposure aggravates DSS-induced colitis in mice by interfering with Akkermansia muciniphila and tryptophan metabolism.

Journal of environmental sciences (China), 161:189-200.

The global incidence of inflammatory bowel disease (IBD) has been escalating. Recent studies have identified co-exposure to polystyrene nanoplastics (PSNP) and triclosan (TCS), two prevalent environmental pollutants, as emerging risk factors for IBD. However, the molecular mechanisms contributing to its deteriorative effect remain elusive. To explore the mechanisms, we conducted an integrative analysis of metagenomic and metabolomic data in a mouse model of colitis induced by dextran sulfate sodium (DSS) following co-exposure to PSNP and TCS. Results demonstrated that co-exposure to PSNP and TCS significantly exacerbated DSS-induced colitis, as evidenced by elevated disease activity indices and pro-inflammatory cytokine levels. Mechanistically, this aggravation correlated with a marked reduction in Akkermansia muciniphila abundance, which was further associated with the disruption of tryptophan metabolism. Specifically, the disruption of this metabolic pathway led to decreased production of two key tryptophan-derived metabolites: indole acetic acid (IAA) and indole acetamide (IAM). In-vitro experiments confirmed that co-exposure to PSNP and TCS inhibited the growth of A. muciniphila rather than affecting the integrity of intestinal epithelial cells. Additionally, IAA and IAM reduced inflammatory cytokine secretion in THP-1 cells. These findings suggest that the reduction in A. muciniphila abundance might decrease the production of IAA and IAM by disrupting tryptophan metabolism. This disruption ultimately contributes to the inflammatory response induced by co-exposure to PSNP and TCS. Our study offers a novel insight into microbiota-host interactions and potential therapeutic targets for intestinal disease.

RevDate: 2025-12-29
CmpDate: 2025-12-29

Geraerts M, Gombeer S, Nebesse C, et al (2025)

A wide diversity of viruses detected in African mammals involved in the wild meat supply chain.

PLoS pathogens, 21(12):e1013643.

The processes involved in acquiring, trading, preparing, and consuming wild meat pose significant risks for the emergence of zoonotic infectious diseases. Several major viral outbreaks have been directly linked to the wild meat supply chain, yet our knowledge of the virome of many mammals involved in this chain remains limited and disproportionately focused on certain mammalian taxa and pathogens. Here, we present the findings of a metagenomic viral screening of 101 mammalian specimens belonging to 28 wild African species and one domesticated species, all traded for their meat. The study focuses on tissue and swab samples collected in various regions in the Democratic Republic of the Congo and in Brussels, Belgium. A total of sixteen virus strains were detected, belonging to the families Arteriviridae, Retroviridae and Sedoreoviridae (primates), Picobirnaviridae (primates and rodents), Picornaviridae (rodents), Hepadnaviridae (hyrax), Orthoherpesviridae (artiodactylid and carnivore) and Spinareoviridae (carnivore). Several strains were detected in mammalian hosts for the first time, expanding their host range and genetic diversity. Of note is the presence of viruses genetically related to recognised zoonotic pathogens, i.e., human picobirnavirus (Orthopicobirnavirus hominis) (primates and rodents), simian foamy viruses (Simiispumavirus) (primates), and rotavirus A (Rotavirus alphagastroenteritidis) (primates). The presence of these viruses in primates is concerning as non-human primates are phylogenetically closely related to humans, which can facilitate interspecies viral transmission. These findings underscore the high diversity of mammalian viruses and the potential risk of human infection through cross-species transmission during close interactions with wildlife in the wild meat supply chain.

RevDate: 2025-12-29

Chen X, Sheng Y, Wang G, et al (2025)

Deciphering Sulfur-Based Denitrification in Confined Alluvial-Lacustrine Aquifers through Multi-isotope ([15]N, [34]S, [13]C, and [18]O) and Metagenomic Analyses.

Environmental science & technology [Epub ahead of print].

Sulfur-based denitrification offers unique advantages in nitrogen removal in low-carbon environments. Despite its importance, the prevalence, controlling factors, and microbial mediators of sulfur-based denitrification in aquifers remain largely unknown. By integrating multiple stable isotopes and genome-resolved metagenomics, we investigated the coupling of nitrogen and sulfur cycling along the groundwater flowpath from unconfined to confined alluvial-lacustrine aquifers in the Poyang Lake Plain, China. Isotope (i.e., δ[15]N-NO3[-], δ[18]O-NO3[-], δ[34]S-SO4[2-], δ[18]O-SO4[2-], δ[18]O-H2O, δ[13]C-DIC, and δ[13]C-DOC) fractionation and hydrogeochemical trends indicated a transition from nitrification-dominated processes in the unconfined aquifer to denitrification, sulfide oxidation, and sulfate reduction in the confined aquifer. A significant negative correlation between δ[34]S-SO4[2-] and δ[15]N-NO3[-], along with a significant positive correlation between genes involved in sulfide oxidation and nitrate reduction, suggested cryptic coupling between sulfide oxidation and nitrate reduction. Sulfur-autotrophic denitrifying metagenome assembly genomes encoding Calvin-Benson-Bassham carbon fixation and rTCA pathways, such as Burkholderiales, Rhodocyclaceae, and Sulfurimonas, were enriched in the confined aquifer and co-occurred with sulfate reducers (Pseudomonadota and Desulfobacterota). These results highlighted sulfur cycling consortia when sulfate was reduced to facilitate removal of nitrate via sulfur-based denitrification. These findings reveal the role of cryptic sulfur cycling in sustaining denitrification in aquifers, offering new insights into nitrogen-sulfur interactions in subsurface ecosystems.

RevDate: 2025-12-29

Oso TA, Ahmed MM, Okesanya OJ, et al (2025)

Exploring the gut-brain-microbiome axis in Alzheimer's disease: Integrating metagenomics, metabolomics, and artificial intelligence for next-generation biomarker discovery.

Journal of Alzheimer's disease : JAD [Epub ahead of print].

Alzheimer's disease (AD), a progressive neurodegenerative disorder, is increasingly understood as a multifactorial condition influenced by systemic and environmental factors beyond the central nervous system. A growing body of evidence shows that the gut-brain-microbiome axis (GBMA), a complex bidirectional communication network, is involved in neural, endocrine, immune, and metabolic pathways in AD pathogenesis. This narrative review synthesizes emerging insights into the role of gut microbiota dysbiosis in promoting neuroinflammation, amyloid-β aggregation, blood-brain barrier disruption, and cognitive decline. We explored recent advancements in metagenomics and metabolomics for profiling microbial communities and their functional metabolites linked to AD. Alterations in microbe-derived compounds, such as short-chain fatty acids and tryptophan metabolites, influence neurodevelopment, glial activation, and mitochondrial dysfunction. Multi-omics integration, enhanced by artificial intelligence (AI), enables precise biomarker discovery, patient stratification, and the development of personalized therapeutic strategies. Translational opportunities include microbiome-based diagnostics, probiotic therapy, and stratified interventions. However, clinical translation faces challenges such as methodological heterogeneity, inter-individual microbiome variation, data governance issues, and algorithmic bias. We emphasize the need for diverse reference panels, longitudinal multimodal cohorts, and shared AI-ready datasets to enhance the reproducibility and global equity of research. Strategic investment in integrative, ethically governed, and interdisciplinary approaches is essential to unlock the full therapeutic and diagnostic potential of GBMA in AD.

RevDate: 2025-12-29

Petersen NP, Le M, Renevey A, et al (2025)

ViMOP: A user-friendly and field-applicable pipeline for untargeted viral genome nanopore sequencing.

Bioinformatics (Oxford, England) pii:8406920 [Epub ahead of print].

MOTIVATION: Untargeted, also known as metagenomic, nanopore sequencing is a powerful tool for virus genomic surveillance, particularly in resource-limited settings and when paired with the portability of the MinION device (Oxford Nanopore Technologies, ONT). However, a major bottleneck for global access is the absence of a user-friendly software capable of efficiently analyzing untargeted nanopore sequencing data to generate high-quality consensus genomes.

RESULTS: We share ViMOP, a pipeline built on our long-term experience in nanopore field sequencing. The pipeline emphasizes field user-friendliness, flexibility and versatility to analyze reads generated directly from human clinical samples. The software assembles de novo contigs, matches contigs to known viral references and uses them to assemble consensus genomes. Executed with a single Nextflow command or via the EPI2ME Desktop interface (ONT), results are summarized in an HTML report. ViMOP, through its user-centered design, lowers the barrier to high-quality virus genome reconstruction and advances capacity for genomic surveillance.

AVAILABILITY: ViMOP is freely available for non-commercial use (https://github.com/opr-group-bnitm/vimop and https://zenodo.org/records/17913089), along with the associated database (https://zenodo.org/records/17652512), the scripts used to generate it (https://zenodo.org/records/17632662) and benchmarking code (https://zenodo.org/records/17633185).

SUPPLEMENTARY INFORMATION: Supplementary data are available at Bioinformatics online.

RevDate: 2025-12-29

Asai N, Igarashi Y, Miyazaki N, et al (2025)

Complete genome sequence of a new Mycobacterium sp. nov. from a compromised Japanese individual.

Microbiology resource announcements [Epub ahead of print].

Here, we report the complete genome sequencing of a new Mycobacterium sp. nov. from a compromised Japanese individual with hematological malignancy. This will represent the essential data for future phylogenetic and comparative genome studies and will be useful for better understanding the evolution of the pathogen.

RevDate: 2025-12-29
CmpDate: 2025-12-29

Duncan A, Koon W, Sidorczuk K, et al (2025)

Detecting signatures underlying the composition of biological data.

Nucleic acids research, 53(22):.

Biological compositional data is inherently multidimensional and therefore difficult to visualize and interpret. To allow for the automatic decomposition of large compositional data and to capture gradients in co-occurring features, called signatures, we developed a new software package 'cvaNMF'. Our benchmarks on synthetic data show the effectiveness of cross-validation and our novel signature-similarity method to identify a suitable decomposition using non-negative matrix factorization (NMF). This software provides a complete set of tools to identify and visualize biologically informative signatures which we demonstrate in a wide range of microbial and cellular datasets: 'Enterosignatures' detected in gut metagenomes differentiated human hosts with diverse diseases; five 'terrasignatures' from rhizosphere metagenomes differentiated root- or soil-associated microbiomes, while being refined enough to infer geographic distances between plants. Large-scale data from >13 000 metagenomes representing 25 biomes were decomposed into environmental and host-associated microbiomes based on five newly discovered signatures. Finally, analysis of the cell composition of non-small cell lung cancer samples allowed separation of cancerous and inflamed tissues based on four cell-type signatures.

RevDate: 2025-12-29

Kwon Y, Choi J, Kim SH, et al (2025)

Rice gs3 allele and low-nitrogen conditions enrich rhizosphere microbiota that mitigate methane emissions and promote beneficial crop traits.

The ISME journal pii:8406465 [Epub ahead of print].

Methane emissions from rice paddies represent a critical environmental concern in agriculture. Although genetic strategies for mitigating emissions have gained attention, the specific microbial and molecular mechanisms remain underexplored. Here, we investigated how the gs3 loss-of-function allele in the near-isogenic rice line Milyang360 modulates rhizosphere and endosphere microbial communities under distinct nitrogen regimes. Field experiments revealed that Milyang360 consistently reduced methane emissions compared with its parental line, Saeilmi, particularly under low-nitrogen conditions. Integrated plant transcriptomic and rhizosphere metagenomic analyses, including the reconstruction of Metagenome-Assembled Genomes, demonstrated that the gs3 allele upregulated genes related to root hair elongation or promoting microbial symbiosis. This physiological change limited substrate availability for methanogens and facilitated the colonization by beneficial microorganisms. Consequently, we observed a functional shift in the microbiome, characterized by the enrichment of methanotrophs and nitrogen-fixing bacteria. This microbial restructuring was most prominent under low-nitrogen conditions, indicating a strong genotype by environment interaction. Our findings highlight the gs3 allele's dual role in reducing methane emissions and improving nitrogen use efficiency by recruiting a beneficial microbiome. This study provides a clear mechanistic link between a plant gene and rhizosphere ecology, offering a promising genetic target for developing sustainable, low emission rice cultivars.

RevDate: 2025-12-29
CmpDate: 2025-12-29

Nandi S, Stephens TG, Walsh K, et al (2025)

Shifts in the microbiome and virome are associated with stony coral tissue loss disease (SCTLD).

ISME communications, 5(1):ycaf226.

Stony coral tissue loss disease (SCTLD) is a rapidly spreading lethal coral disease, the etiology of which remains poorly understood. In this study, using deep metagenomic sequencing, we investigated microbial and viral community dynamics associated with SCTLD progression in the Caribbean stony coral Diploria labyrinthiformis. We assembled 264 metagenome-assembled genomes and correlated their abundance with disease phenotypes, which revealed significant shifts in both the prokaryotic microbiome and virome. Our results provide clear evidence of microbial destabilization in diseased corals, suggesting that microbial dysbiosis is an outcome of SCTLD progression. We identified DNA viruses in our dataset that increase in abundance in SCTLD-affected corals and are present in existing coral data from other Caribbean regions. In addition, we identified the first putative instance of asymptomatic/resistant SCTLD-affected corals. These are apparently healthy colonies that share the viral profile of diseased individuals. However, these colonies contain a different prokaryotic microbiome than do diseased corals, suggesting microbe-induced resilience (i.e. beneficial microbiome) to SCTLD. Finally, utilizing differential abundance analysis and gene inventories, we propose a mechanistic model of SCTLD progression, in which viral dynamics may contribute to microbiome collapse. These findings provide novel insights into SCTLD pathogenesis and offer consistent molecular signals of disease across diverse geographic sites, presenting new opportunities for disease monitoring and mitigation.

RevDate: 2025-12-29
CmpDate: 2025-12-29

Mulec J, Pašić L, A Oarga-Mulec (2025)

Metabolic traits of sediment bacteria in karst caves in the light of environmental changes.

Frontiers in microbiology, 16:1724116.

INTRODUCTION: Karst subterranean systems are vulnerable ecosystems that have not yet been studied adequately at the microbial functional level. Cave sediments deposited over different time periods host diverse microbial communities that play a critical role in nutrient cycling and pollutant degradation.

METHODS: In this study, we investigated microbial diversity and metabolic capacity in recently deposited alluvial sediments and an ancient palaeo-river deposit in a karst cave system. Using 16S rRNA gene amplicon metagenomic analysis, community-level physiological profiling (CLPP), and chemical characteristics of the environment, the influence of key environmental factors on microbial community composition and substrate degradation, concentrating particularly upon sediment age, oxygen availability, and temperature, was assessed.

RESULTS: The results showed different microbiome compositions and metabolic characteristics between sites. The old alluvial sediment exhibited low taxonomic and functional diversity, accompanied by elevated heavy-metal concentrations, suggesting that sediment age might act as a geochemical filter, limiting microbial function. In contrast, a periodically flooded site showed high metabolic versatility and taxonomic diversity, emphasizing the ecological role of hydrological pulses in maintaining functional microbial diversity. CLPP metrics linked community structure to functional potential, revealing adaptive traits in key taxa such as Polaromonas, Methylibium, and Beggiatoa.

DISCUSSION: These results demonstrated the value of integrating functional and taxonomic approaches in subsurface environments and provide insights into microbial resilience, biogeochemical processes, and the potential for applied environmental use.

RevDate: 2025-12-29
CmpDate: 2025-12-29

Ruiz-Blas F, Bartholomäus A, Henny C, et al (2025)

Cryptic sulfur cycling in the deep biosphere of ferruginous Lake Towuti, Indonesia.

Frontiers in microbiology, 16:1725877.

Lake Towuti, Sulawesi, Indonesia is an ancient tectonic lake, exhibiting iron-rich, sulfate-poor anoxic deep waters. Temporal variations in water column stratification led to sediment accumulation under variable redox conditions. Such ferruginous settings make Lake Towuti an ideal study site to evaluate how a cryptic sulfur cycle could possibly operate under a scarcity of sulfate and abundance of iron minerals, similar to Earth's primitive oceans. Here, we integrate downcore profiles for pore water geochemistry, reactive iron mineralogy, and bulk sediment elemental composition with microbial cell counts, sulfate reduction rates, 16S rRNA genes and metagenomes to resolve microbial sulfur transformations down to 15 m below lake floor (mblf). Sulfate concentrations and reduction rates dropped within the upper mblf, while pore water ferrous iron increased to its highest concentration down to 3 mblf. Any microbially-produced sulfide precipitated as reduced inorganic sulfur in the sediment, apparently forming authigenic millerite (NiS) during burial. The decrease in cell densities tracked the decline in electron acceptors in pore waters with depth. From 3 to 10 mblf, low but sustained sulfate reduction rates were observed with intermittent presence of nitrate in pore water and increased goethite in the sediment, both acting as potential oxidants of sulfur intermediates. A subsequent re-increase in pore water sulfate occurred in parallel with syntrophic fermentation of volatile fatty acids. Consistent with geochemical evolution, the taxonomic diversity of microbial populations shifted from a bacterial assemblage near the surface to selective but prevailing Bathyarchaeia down to 15 mblf. The corresponding metagenome-assembled genomes predicted metabolic potential for complete sulfate reduction (aprAB, dsrAB) in Thermodesulfovibrionia, whereas Desulfobacterota (incl. Geobacterales, Desulfuromonadales, Syntrophales) and Aminicenantia exhibited versatility in reducing iron, nitrate (narG, napA), nitrite (nirS, nrfA) and sulfate (dsrAB, asrA). By contrast, Bathyarchaeia were predicted to disproportionate sulfur to polysulfides and reduce ferredoxin via electron bifurcation (hyd I-II, sudA, dsrC, dsrE) to fuel a Wood-Ljungdahl pathway, defining homoacetogenesis as terminal electron sink. Together, these mineralogical, geochemical, and metagenomic features provide evidence for a spatially confined but active cryptic sulfur cycle with tight coupling between reduction of mineral ferric iron and intermittent pore water nitrate to syntrophic and lithotrophic (homo)acetogenesis.

RevDate: 2025-12-29
CmpDate: 2025-12-29

Zhao X, Qiao J, Wang Y, et al (2025)

Shotgun metagenomics reveals antibiotic resistome dynamics and metabolic specialization in fungal-dominated microbiomes.

Frontiers in microbiology, 16:1626799.

BACKGROUND: Metagenomics offers a culture-independent framework for comprehensively characterizing microbial communities by directly extracting and sequencing DNA from environmental samples. In this study, we employed high-throughput metagenomic sequencing to explore microbial communities inhabiting fungal-rich environments, emphasizing taxonomic composition, functional potential, and antibiotic resistance gene (ARG) dynamics.

METHODS: Six samples from two distinct groups (HFJ and QFJ) were subjected to Illumina-based shotgun sequencing, followed by rigorous quality control, taxonomic classification, KEGG-based functional annotation, and ARG identification via the CARD database. Comparative analysis revealed stark contrasts between the two groups.

RESULTS: HFJ samples were dominated by eukaryotic taxa, particularly Saccharomyces cerevisiae, and exhibited elevated carbohydrate metabolism, aligning with the ecological role of fermentative fungi. Conversely, QFJ samples displayed higher bacterial diversity, particularly Firmicutes and Proteobacteria, and were enriched in lipid and amino acid metabolism pathways. Striking differences were also observed in ARG profiles. QFJ samples harbored greater ARG abundance, particularly genes conferring resistance to beta-lactams, aminoglycosides, and tetracyclines, indicating higher resistance potential and possible horizontal gene transfer activity.

CONCLUSION: Our results reveal distinct microbial, functional and resistome profiles in fungal-rich versus bacterial-rich fermentation environments. Fungal dominance correlated with lower bacterial diversity and a reduced abundance of certain ARGs, whereas bacterial-rich samples exhibited higher diversity and ARG prevalence. These correlations generate the hypothesis that fungal dominance may suppress bacterial growth or ARG dissemination; however, causal relationships cannot be inferred from our cross-sectional data. The study highlights the potential of metagenomic surveillance to elucidate ecological niches that influence bacterial diversity and resistance dynamics.

RevDate: 2025-12-29
CmpDate: 2025-12-29

Li P, Wang M, Zhang H, et al (2025)

Integrative metagenomic and metabolomic profiling identifies gut microbial and metabolite signatures associated with lymph node metastasis in pancreatic cancer.

Frontiers in microbiology, 16:1706084.

BACKGROUND: Lymph node metastasis (LNM) is a prognostic factor in pancreatic cancer. The association between the gut microbiota and LNM remains unexplored. This study aimed to characterize the gut microbiota and metabolomic profiles associated with LNM and to investigate their potential as predictive biomarkers.

METHODS: Fecal samples from pancreatic cancer patients undergoing surgery were analyzed using metagenomic sequencing and untargeted metabolomics. The patients were categorized into LNM and non-LNM (NLNM) groups. Differential microbiome taxa were analyzed using the DESeq2 package. Random forest predictive models were developed based on metagenomic and metabolomic data, with performance assessed using leave-one-out cross-validation (LOOCV).

RESULTS: A total of 26 patients with LNM and 29 patients without LNM were included. Principal coordinates analysis (PCoA) revealed significant differences in microbiota composition between the two groups (Anosim, p = 0.047). The absolute counts of Ruminococcus gnavus and Blautia wexlera were significantly decreased in LNM. Tryptophan-derived metabolites, indole-3-lactic acid (3-ILA) and indole-3-acrylic acid (3-IA), were downregulated in LNM. Functional pathway analysis showed downregulation of tryptophan metabolism in LNM, while cancer-related pathways were upregulated. Correlation analysis revealed a significant positive association between Ruminococcus gnavus and 3-ILA/3-IA levels. Moreover, Ruminococcus gnavus was positively correlated with CD8[+] T cells. Predictive models based on the gut microbiota and metabolites distinguished LNM from NLNM, with AUC values of 0.854 and 0.940, respectively.

CONCLUSION: The gut microbiota and metabolites exhibit significant alterations during lymph node metastasis in pancreatic cancer, especially Ruminococcus gnavus, Blautia wexlera, and tryptophan metabolites (3-ILA and 3-IA). Gut microbial and metabolite signatures may serve as potential non-invasive biomarkers for predicting LNM in pancreatic cancer. Further functional validation is required to determine whether and how the gut microbiota and metabolites may mediate lymph node metastasis.

RevDate: 2025-12-29
CmpDate: 2025-12-29

Brenes-Guillén L, Vidaurre-Barahona D, Agüero K, et al (2025)

Novel diversity of Anaerolineae and Tepidiformia recovered from metagenomes of thermal microbial mats in Costa Rica.

Frontiers in microbiology, 16:1693256.

Tropical thermal and mineral springs are ideal for studying microbial life in extreme environments, their microbial diversity, and functional profiles. In this study, we investigated the abundance and genomic diversity of the phylum Chloroflexota in microbial mats from 33 thermal and acidic springs across Costa Rica using 16S rRNA gene amplicon sequencing and shotgun metagenomics. Our results demonstrated that pH and temperature are the main environmental drivers shaping the abundance and diversity of Chloroflexota. Acidic conditions favored the presence of Ktedonobacteria and the candidate division AD3, while thermal environments were dominated by unclassified Anaerolineae. From a subset of thermal springs, we reconstructed 72 metagenome-assembled genomes (MAGs), many of which represent previously uncharacterized lineages. Comparative genomic analyses revealed two novel families and seven new genera within Anaerolineae and a distinct lineage within Tepidiformia. We proposed the following names: Ca. Sittenfelaceae, Ca. Mariellaceae, and Ca. Tepidiforma platanarica. Functional annotation of Anaerolineae and Tepidiformia MAGs suggested a degree of functional redundancy. Genes associated with methanogenesis, dissimilatory nitrate reduction, sulfur metabolism, and methylotrophy were detected, while genes involved in photosynthesis, nitrogen fixation, and nitrification were absent. Unique gene clusters were identified in each family, and interestingly, 23% of these unique genes were of unknown function, highlighting the unexplored genetic potential of these organisms. Canonical correspondence analysis (CCA) revealed that temperature significantly influences the microdiversity of Anaerolineae. Despite their taxonomic novelty, these lineages exhibit strong functional redundancy across major metabolic pathways, where overlapping metabolic capabilities may confer stability under fluctuating conditions and support the persistence of diverse Chloroflexota populations. This study provides the first genomic dataset of Chloroflexota from Central American geothermal environments and highlights tropical geothermal springs as reservoirs of novel microbial diversity and functional potential.

RevDate: 2025-12-29
CmpDate: 2025-12-29

Ng C, Abrazaldo J, de Vera P, et al (2025)

Antibiotic Resistance in the Philippines: Environmental Reservoirs, Spillovers, and One-Health Research Gaps.

Frontiers in microbiology, 16:1711400.

Antimicrobial resistance (AMR) remains a major health threat in the Philippines, where high antimicrobial use, intensive aquaculture, and recurrent typhoon-driven flooding and monsoon seasons shape distinctive transmission pathways. This narrative review synthesizes published Philippine data across clinical, agriculture, and environmental sectors to map evidence and gaps relevant to policy. Clinically, vancomycin resistant Enterococcus faecium is 23%, Klebsiella pneumonia shows ∼15% carbapenem resistance, and Escherichia coli resistance to third generation cephalosporins (3GC) and fluoroquinolone are ∼43 and ∼46%, respectively. In food animals, ceftriaxone resistance in non-typhoidal Salmonella (NTS) increased from ∼8% (2010) to ∼43% (2020s), with ciprofloxacin resistance between 14 and 23%. Environmental studies report extended spectrum beta-lactamase (ESBL)-producing E. coli in Manila estuaries and multiple antibiotic resistance (MAR) indices of up to 0.15 in tributaries. Hospital sewage and nearby rivers have yielded carbapenemase-producing Enterobacterales (CPE) bearing bla NDM/bla KPC in clinically relevant lineages (e.g., E. coli CC10, K. pneumoniae ST147). Cross sector comparisons remain constrained by method heterogeneity and data gaps. To operationalize One-Health monitoring, we propose (i) a two window surveillance design: late dry baseline and 24-72 h post-storm flood pulses; and (ii) a two tier analytics model: Tier 1 HT-qPCR ARG/MGE panels at regional hubs for rapid screening, and Tier 2 metagenomics/isolate whole genome sequencing (WGS) at national hubs for source attribution and plasmid tracking. We translate these findings into a modular AMR risk assessment toolkit to prioritize surveillance and targeted interventions.

RevDate: 2025-12-29
CmpDate: 2025-12-29

Zhao Z, Zhang Y, Fu J, et al (2025)

Metagenomic next-generation sequencing for cryptococcal meningitis diagnosis: a single-center experience.

Frontiers in cellular and infection microbiology, 15:1626290.

PURPOSE: Cryptococcal meningitis (CM) is a devastating central nervous system infection with substantial mortality, particularly when diagnosis is delayed. This study aims to evaluate the diagnostic performance of metagenomic next-generation sequencing (mNGS) for CM detection in comparison with conventional tests.

METHODS: We enrolled 23 consecutive patients with suspected CM at a tertiary center. All patients met a composite reference standard (CRS) based on CSF cryptococcal antigen (CrAg), CSF/sterile-site culture for Cryptococcus, or CNS histopathology; mNGS was excluded from the CRS. Primary outcomes were CRS-based sensitivity (computed only among CRS-positive patients who underwent each assay) and turnaround times (TATs); pairwise agreement metrics (PPA/NPA) between mNGS and conventional assays were estimated in co-tested subsets.

RESULTS: mNGS identified Cryptococcus in 18/23 (78.3%) cases and detected viral co-pathogens (EBV/CMV/HIV-1) in 5 patients. CRS-based sensitivities were: CrAg LFA (CSF) 83.3% (5/6), Alcian blue 72.7% (16/22), India ink 50.0% (3/6), and CSF culture 66.7% (8/12). Pairwise agreement favored mNGS against culture and CrAg (e.g., PPA 100% vs culture 8/8] and vs CSF CrAg [5/5]), with limited NPA where denominators were small. Median (IQR) TATs were 0.5 (0.5-0.5) days for CrAg LFA, 1 (0.5-1) day for India ink, 5 (3-8) days for first positive culture, and 2 (1-4) days for mNGS.

CONCLUSION: CSF mNGS complements CrAg, microscopy, and culture by increasing Cryptococcus detection and revealing mixed infections, with particular utility in atypical, pretreated, or complex hosts. Larger studies are warranted to validate clinical utility and define optimal integration with existing workflows.

RevDate: 2025-12-29
CmpDate: 2025-12-29

Dai L, Mao L, Shi W, et al (2025)

A circovirus-like genomic sequence with unique architecture and phylogenetic relatedness to human-linked viral lineages.

Frontiers in cellular and infection microbiology, 15:1698076.

Circoviruses are small viruses in the Circoviridae family that associate with a wide spectrum of disease from asymptomatic to lethal in different animal. In the current study, a novel swine-origin circovirus with unique genomic architecture was determined from both oropharyngeal swabs and fecal samples collected from febrile pigs in Jiangsu Province, China. The complete genome sequence of this novel porcine circovirus strain (named as PCV-CH-ZJ01) was determined to be 2544 nt in length, containing four major open reading frames: Rep1, 498-833 nt, 112 aa; Rep2, 1055-1546 nt, 164 aa; Cap1, 1756-2055 nt, 100 aa; Cap2, 2089-2529 nt, 147 aa. Comparative sequence analysis revealed differential conservation patterns across viral components and shared low amino acid similarity with other strains on the whole. Phylogenetic trees based the whole genome and amino acid of two Reps revealed that PCV-CH-ZJ01 belonged to circovirus and clustered with human and rhesus macaque circovirus strains, which raised the concerns of potential cross-species transmission between swine and human.

RevDate: 2025-12-29
CmpDate: 2025-12-29

Wróbel M, Gawryś R, Tereba A, et al (2025)

Dependence of Bacterial OTUs on Selected Features of Beaver Ponds.

Ecology and evolution, 15(12):e72790.

Ponds created by beavers represent unique aquatic ecosystems that influence hydrological, chemical and biological conditions, including the microbiology of the water. The activity of these animals promotes biodiversity and water purification processes, but can also lead to the accumulation of pollutants. Water retention in beaver ponds promotes the development of bacteria and other microorganisms that play an important role in biogeochemical cycles. Long-term water stagnation can lead to anaerobic conditions and the formation of toxic compounds, which in turn can limit the diversity of benthic organisms. Beavers play a key role in shaping these habitats, and microbiological studies of their reservoirs provide a better understanding of their impact on aquatic ecosystems, self-purification processes and potential biological threats. Metagenomic analysis revealed the presence of 365 bacterial species in water and sediment samples, belonging to 174 genera and 83 families. 83 operational taxonomic units (OTUs) were identified, 62 of which were present in both water and sediments. Although the overall OTU composition was similar in both environments, greater variability was observed in the sediments. The statistical differences in OTU distribution between water and sediments were confirmed using the Wilcoxon test.

RevDate: 2025-12-29
CmpDate: 2025-12-29

Wu X, Shao T, Huang Y, et al (2025)

Effects of dietary nisin supplementation on the growth performance, serum biochemistry, digestive enzyme activities, intestinal morphology, and intestinal microbiota in rabbits.

Frontiers in veterinary science, 12:1726365.

INTRODUCTION: This study evaluated the impact of dietary supplementation with varying doses of nisin (NI) on the growth performance, serum biochemical parameters, intestinal digestive enzyme activity, short-chain fatty acid (SCFA) profiles, mucosal morphology, and the cecal microbiota composition in rabbits.

METHODS: Healthy female New Zealand white rabbits (5 weeks old; n = 90) of comparable body weight were randomly allocated to five groups: a positive control (PC) group receiving a basal diet supplemented with kitasamycin (300 mg/kg), three NI groups supplemented with nisin at 600 (NI600), 800 (NI800), or 1,000 (NI1000) mg/kg, and a negative control (NC) group receiving the basal diet without additives. Each treatment was comprised of three replicates (n = 6 per replicate), and the trial lasted 42 days.

RESULTS: The results showed that the rabbits administered NI displayed significantly enhanced final body weights as compared to the NC group (P < 0.05), with a dose-dependent effect. Notably, the NI800 and NI1000 groups exhibited a superior average daily gain (ADG) and average daily feed intake (ADFI). Serum analyses showed improved lipid profiles and elevated antioxidant enzyme activities concomitant with reduced lipid peroxidation in the NI-supplemented groups. Enzymatic assays indicated elevated duodenal a-amylase activity in the NI800 group as compared to the PC (P < 0.05) and enhanced ileal trypsin activity in the NI800 as compared to NI1000 and PC (P < 0.05). Histological evaluation confirmed that the NI800 group displayed optimal intestinal villi morphology, characterized by increased density, height, and structural integrity relative to the PC and NC controls. Metagenomic analysis of the cecal microbiota further revealed dose-dependent shifts in the diversity and composition of the microbiota, with the NI800 group exhibiting pronounced restructuring. Enriched functional pathways in the NI groups, including cofactor/vitamin metabolism, amino acid biosynthesis, energy homeostasis, and environmental adaptation.

DISCUSSION: Collectively, these findings highlight that NI supplementation enhances digestive efficiency, augments systemic antioxidant defenses, fortifies intestinal barrier function, and modulates microbial ecology and SCFA production, there by promoting growth and metabolic health in rabbits. Nisin, especially at 800 mg/kg, demonstrates significant potential as an antibiotic alternative.

RevDate: 2025-12-29
CmpDate: 2025-12-29

Gopep NS, Odukale-Okuneye AO, Osanakpo IM, et al (2025)

Association Between Gut Microbiota Diversity and Body Mass Index (BMI) in Healthy Young Adults in the United States: Insights Into the Gut-Brain-Metabolic Axis Using the Curated Metagenomic Data.

Cureus, 17(11):e97746.

BACKGROUND: Emerging evidence suggests that gut microbiota diversity plays a critical role in metabolic regulation and may influence body mass index (BMI). However, findings in healthy populations remain inconsistent.

OBJECTIVE: This study aims to determine whether gut microbiota alpha-diversity is associated with BMI among healthy young adults aged 18-39 years in the United States and to explore potential implications for the gut-brain-metabolic axis.

METHODS: This cross-sectional study utilized publicly available metagenomic data from the CuratedMetagenomicData repository. After preprocessing in R version 4.5.0 (R Foundation for Statistical Computing, Vienna, Austria), data were analyzed using Stata version 18 (Released 2023; StataCorp LLC, College Station, TX). Alpha-diversity indices (Shannon, Simpson, and Richness) were computed and examined across BMI categories (normal, overweight, and obese) using one-way analysis of variance (ANOVA) and chi-square tests. Linear regression models were employed to assess associations between BMI and diversity measures, adjusting for age and gender.

RESULTS: Among 147 participants, BMI differed significantly across weight categories (p < 0.001), but no significant association was observed between Shannon diversity and BMI (p = 0.527). Age emerged as the only significant predictor of BMI in adjusted models (p < 0.001).

CONCLUSION: Gut microbial alpha-diversity was not significantly associated with BMI among healthy young adults. Functional microbial characteristics, rather than diversity alone, may better explain variations in metabolic status.

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ESP Quick Facts

ESP Origins

In the early 1990's, Robert Robbins was a faculty member at Johns Hopkins, where he directed the informatics core of GDB — the human gene-mapping database of the international human genome project. To share papers with colleagues around the world, he set up a small paper-sharing section on his personal web page. This small project evolved into The Electronic Scholarly Publishing Project.

ESP Support

In 1995, Robbins became the VP/IT of the Fred Hutchinson Cancer Research Center in Seattle, WA. Soon after arriving in Seattle, Robbins secured funding, through the ELSI component of the US Human Genome Project, to create the original ESP.ORG web site, with the formal goal of providing free, world-wide access to the literature of classical genetics.

ESP Rationale

Although the methods of molecular biology can seem almost magical to the uninitiated, the original techniques of classical genetics are readily appreciated by one and all: cross individuals that differ in some inherited trait, collect all of the progeny, score their attributes, and propose mechanisms to explain the patterns of inheritance observed.

ESP Goal

In reading the early works of classical genetics, one is drawn, almost inexorably, into ever more complex models, until molecular explanations begin to seem both necessary and natural. At that point, the tools for understanding genome research are at hand. Assisting readers reach this point was the original goal of The Electronic Scholarly Publishing Project.

ESP Usage

Usage of the site grew rapidly and has remained high. Faculty began to use the site for their assigned readings. Other on-line publishers, ranging from The New York Times to Nature referenced ESP materials in their own publications. Nobel laureates (e.g., Joshua Lederberg) regularly used the site and even wrote to suggest changes and improvements.

ESP Content

When the site began, no journals were making their early content available in digital format. As a result, ESP was obliged to digitize classic literature before it could be made available. For many important papers — such as Mendel's original paper or the first genetic map — ESP had to produce entirely new typeset versions of the works, if they were to be available in a high-quality format.

ESP Help

Early support from the DOE component of the Human Genome Project was critically important for getting the ESP project on a firm foundation. Since that funding ended (nearly 20 years ago), the project has been operated as a purely volunteer effort. Anyone wishing to assist in these efforts should send an email to Robbins.

ESP Plans

With the development of methods for adding typeset side notes to PDF files, the ESP project now plans to add annotated versions of some classical papers to its holdings. We also plan to add new reference and pedagogical material. We have already started providing regularly updated, comprehensive bibliographies to the ESP.ORG site.

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Papers in Classical Genetics

The ESP began as an effort to share a handful of key papers from the early days of classical genetics. Now the collection has grown to include hundreds of papers, in full-text format.

Digital Books

Along with papers on classical genetics, ESP offers a collection of full-text digital books, including many works by Darwin and even a collection of poetry — Chicago Poems by Carl Sandburg.

Timelines

ESP now offers a large collection of user-selected side-by-side timelines (e.g., all science vs. all other categories, or arts and culture vs. world history), designed to provide a comparative context for appreciating world events.

Biographies

Biographical information about many key scientists (e.g., Walter Sutton).

Selected Bibliographies

Bibliographies on several topics of potential interest to the ESP community are automatically maintained and generated on the ESP site.

ESP Picks from Around the Web (updated 28 JUL 2024 )