Salmonella is a major zoonotic foodborne pathogen, and antimicrobial resistance (AMR) in Salmonella presents a significant public health challenge. Compared with conventional antimicrobial susceptibility testing (AST), whole-genome sequencing (WGS) provides a more rapid and comprehensive approach to AMR characterization, thereby informing antimicrobial selection and supporting public health surveillance. In this study, Oxford Nanopore Technology (ONT)-based WGS was performed on 1,490 Salmonella isolates collected through nationwide surveillance in Taiwan in 2025. Genotypic resistance inferred from WGS data was compared with phenotypic AST results to assess the performance of ONT-WGS. Overall, WGS-inferred resistance showed high concordance with phenotypic resistance for most antimicrobials. However, major genotype-phenotype discordance was observed, attributed to four categories: (i) breakpoint-dependent classification, (ii) reduced or absent phenotypic expression of resistance genes, (iii) minimum inhibitory concentration (MIC) modulation by ramAp, and (iv) absence of known AMR determinants. Notable discrepancies included tigecycline resistance without known genetic determinants, nalidixic acid resistance linked to ramAp-mediated MIC elevation, and a high prevalence of colistin resistance (35.7%) in S. Enteritidis, with most resistant isolates lacking identifiable AMR determinants. Additionally, a significant proportion of ESBL- and AmpC-producing isolates were classified as susceptible or intermediate to cefotaxime and ceftazidime under CLSI criteria, highlighting the potential for misclassification and treatment failure. These findings demonstrate that ONT-WGS enables accurate and comprehensive AMR characterization by directly identifying resistance determinants and avoiding potential misclassification associated with breakpoint-based AST interpretations. When interpreted appropriately, WGS can support better antimicrobial selection and serve as a valuable alternative to conventional susceptibility testing. Accurate prediction of antimicrobial resistance is essential for appropriate therapy and effective surveillance of Salmonella. However, discordance between genotype-based predictions and phenotypic antimicrobial susceptibility testing (AST) can complicate clinical interpretation. In this nationwide study of 1,490 Salmonella isolates, we show that Oxford Nanopore Technology-based whole-genome sequencing (ONT-WGS) provides rapid and comprehensive detection of antimicrobial resistance determinants with high concordance to phenotypic AST. We further identify four major mechanisms underlying genotype-phenotype discordance, including breakpoint-dependent classification, reduced or absent phenotypic expression of resistance genes, minimum inhibitory concentration (MIC) modulation by ramAp, and the absence of known AMR determinants. These findings demonstrate how WGS can complement conventional AST, improve interpretation of challenging susceptibility results, and strengthen genomic surveillance of emerging antimicrobial-resistant Salmonella.
Recently, novel nanobiotechnological approaches have been developed to reduce the impact of pollutants during the synthesis of nanoparticles (NPs). To do so, green synthesis methods have now become widely adopted, using biomolecules to produce high-quality, stable nanoparticles. In this work, we employed the K1 toxin produced by S. cerevisiae as a reducing and capping agent for silver nanoparticles. The synthesis of Ag-K1 NPs was carried out using a concentrated protein fraction from the culture medium of S. cerevisiae 42300 containing the secreted K1 toxin. The obtained nanoparticles were characterized using UV-Vis spectroscopy, STEM, EDS, and FTIR, and the antimicrobial efficacy was determined against S. cerevisiae, P. aeruginosa, and B. subtilis. The synthesized NPs showed high antimicrobial efficacy, killing all tested strains; additionally, dose-response modeling suggested differential activity among the nanoparticles. This work reports, for the first time, the bio-assisted synthesis and antimicrobial characterization of silver nanoparticles associated with K1 killer toxin-containing protein fractions.
The emergence and spread of multidrug-resistant Enterobacteriaceae (MDR-E) in community and hospital settings constitute a major public health challenge, particularly in densely populated urban areas. Although antimicrobial resistance among clinical isolates has been repeatedly studied, resistance profiles of intestinal carriage strains remain scarcely reported, despite their potential role as important reservoirs of resistance genes. An integrated approach including both carriage and clinical isolates is therefore necessary to better understand antimicrobial resistance dynamics. This study is aimed at comparing the antibiotic resistance profiles of Enterobacteriaceae isolated from intestinal carriage in outpatients and those from clinical infections. A total of 196 strains, including Escherichia coli, Klebsiella pneumoniae, and Enterobacter cloacae, were analyzed, comprising 60 carriage isolates and 136 clinical isolates. Antimicrobial susceptibility testing was performed using the disk diffusion method according to CA-SFM guidelines. Resistance profiles were classified as multidrug-resistant (MDR), extensively drug-resistant (XDR), or pandrug-resistant (PDR) based on the criteria of Magiorakos et al., and the multiple antibiotic resistance (MAR) index was calculated. Overall, a high prevalence of MDR strains was observed (86.73%) in both groups, with distinct resistance patterns depending on the source. Carriage strains showed higher resistance to cephalosporins, fluoroquinolones, monobactams, tetracyclines, and sulfonamides (MAR index: 0.76 ± 0.09), whereas clinical strains exhibited increased resistance to aminoglycosides and carbapenems (MAR index: 0.65 ± 0.18). XDR (6.12%) and PDR (1.02%) phenotypes were mainly identified among clinical isolates. These findings emphasize the role of intestinal carriage as a reservoir of antimicrobial resistance and support the need for integrated community-hospital surveillance strategies in urban settings.
Antimicrobial resistance poses a looming global health challenge impacting human, animal, and environmental health sectors, with horizontal gene transfer (HGT) playing a critical role in the dissemination of resistance genes among bacteria. To address this challenge effectively, a comprehensive strategy encompassing animal health and environmental management is essential. Therefore, this study aims to investigate the efficacy of phytochemicals on the conjugative transfer of an antimicrobial resistance gene, bla TEM, between multidrug-resistant Salmonella Heidelberg, a major food-borne pathogen in poultry and commensal Escherichia coli. The effect of four phytochemicals, namely trans-cinnamaldehyde (TC), carvacrol (CR), beta-resorcylic acid (BR), and caprylic acid (CA) were determined across diverse environments, including bacteriological broth, chicken manure, and water. Further, the efficacy of in-feed supplementation of TC, CR and TC-CR combinations in reducing HGT frequency in broiler chicken ceca ex vivo was also studied. The HGT frequency was calculated as the ratio of the number of transconjugant CFU per mL to the number of recipient CFU per mL. Exposure to CR and BR reduced transconjugant counts and HGT frequency in broth and chicken manure, while CR and CA showed similar effects in water (p < 0.05). Additionally, in-feed supplementation 0.5% TC decreased HGT frequency in chicken ceca ex vivo (p < 0.05). There was also a significant reduction in the transcription of conjugation genes in Salmonella treated with phytochemicals (p < 0.05). These findings suggest the potential of phytochemicals to mitigate the spread of bla TEM in the poultry production continuum, although additional studies in a large number of samples, including chickens, are imperative to validate these results.
Green synthesis of silver nanoparticles (AgNPs) using plant extracts offers an eco-friendly and sustainable strategy for developing multifunctional nanomaterials with biomedical applications. This study aimed to biosynthesize AgNPs using Syzygium aromaticum (clove) extract and evaluate their physicochemical characteristics and antioxidant, anti-inflammatory, antiviral, anticancer, and antibacterial activities. AgNPs were biosynthesized using clove extract and characterized by Ultraviolet-Visible Spectroscopy (UV-Vis), X-ray Diffraction (XRD), Transmission Electron Microscopy (TEM), and Energy-Dispersive X-ray Spectroscopy (EDX). Antioxidant activity was determined by IC₅₀ analysis, while anti-inflammatory potential was assessed using hemolysis protection and human red blood cell (HRBC) membrane stabilization assays. Antiviral activity was evaluated against hepatitis A virus (HAV), Coxsackievirus B4 (COXB4), and herpes simplex virus type 1 (HSV-1). Cytotoxicity was assessed in human colorectal carcinoma (CaCO₂) and normal lung fibroblast (Wi-38) cells. Antibacterial activity was tested against multidrug-resistant (MDR) bacterial strains, and quantitative molecular analyses were performed to determine the expression of bacterial virulence genes and cancer-related genes. All reported values represent the mean of three independent experiments. The synthesized AgNPs were predominantly spherical, well dispersed, and had an average particle size of approximately 9.5 nm. XRD confirmed a crystalline face-centered cubic (fcc) structure, while UV-Vis analysis showed a characteristic surface plasmon resonance peak at 437 nm, confirming successful nanoparticle formation. The AgNPs exhibited strong antioxidant activity with an IC₅₀ of approximately 13.5 μg/mL and demonstrated significant anti-inflammatory activity through membrane stabilization and hemolysis protection. Antiviral assays showed marked inhibition of HAV, COXB4, and HSV-1 at 250 μg/mL, with the highest inhibition observed against HAV (74%). Cytotoxicity studies revealed selective anticancer activity, with an IC₅₀ of approximately 137 μg/mL against CaCO₂ cells compared with 387 μg/mL for Wi-38 cells. Broad-spectrum antibacterial activity was observed against MDR Staphylococcus aureus (ATCC 27217), Staphylococcus haemolyticus (ATCC 29970), Escherichia coli (BAA-197), and Klebsiella pneumoniae (BAA-1705). Molecular analyses demonstrated significant suppression of bacterial virulence genes, including luxS in E. coli (70%), fnbA-a and cna in S. aureus (74%), fnbA-h in S. haemolyticus (77%), and rmpA in K. pneumoniae (81%) at AgNP concentrations of 10 μg/mL for Gram-positive and 12.5 μg/mL for Gram-negative strains. In CaCO₂ cells, AgNP treatment significantly downregulated c-MYC (0.49-fold), K-RAS (0.67-fold), and BCL2 (0.78-fold; p < 0.01), while significantly upregulating BAX (1.74-fold). Clove-mediated AgNPs demonstrated potent multifunctional biological activities, including antioxidant, anti-inflammatory, antiviral, selective anticancer, and antibacterial effects, accompanied by modulation of cancer-associated and bacterial virulence genes. These findings highlight the therapeutic potential of biosynthesized AgNPs as promising candidates for the development of novel antimicrobial and anticancer nanomedicines.
CuO-PQ nanocomposites with 0.5 (CP 0.5), 1.0 (CP 1.0) and 1.5 wt% (CP 1.5) of PQ were prepared using a wet impregnation route and assessed for sunlight driven dye degradation and antibacterial activity. Powder X-ray diffraction confirmed pristine monoclinic CuO together with characteristic PQ reflections in the composites. UV-vis diffuse reflectance spectroscopy showed progressive band gap narrowing from 2.32 eV for CuO to 1.75, 1.68 and 1.63 eV for CP 0.5, CP 1.0 and CP 1.5, respectively, thereby increasing visible light absorption. CP 0.5 exhibited the highest photocatalytic performance, increasing the apparent rate constants for rhodamine 6G, methylene blue and safranine from 1.80 × 10-3, 2.40 × 10-3 and 3.92 × 10-3 min-1 for bare CuO to 1.03 × 10-2, 5.40 × 10-3 and 1.44 × 10-2 min-1, respectively. In antibacterial tests against Escherichia coli (EC), Pseudomonas aeruginosa (PA) and Staphylococcus aureus (SA), the CuO-PQ composites displayed significantly higher inhibition zones than CuO alone, reaching up to 27 mm for CP 1.0 and CP 1.5 against SA. While PQ has previously been incorporated into photocatalytic systems, its intrinsic antimicrobial activity has, to the best of our knowledge, not been reported earlier. In the current work we show that PQ acts as an antimicrobial active component within CuO-PQ nanocomposites. The improved dual functionality is attributed to interfacial coupling between CuO and PQ, which broadens the visible light response, promotes charge separation and enables more efficient generation of reactive oxygen species (ROS) under sunlight.
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In the established model of classical trained immunity, metabolic and epigenetic hubs serve as central integrators of innate memory. While typically associated with proinflammatory reprogramming, the regulation of autophagy and cellular proteostasis remains essential for guiding macrophage differentiation and ensuring efficient pathogen clearance without excessive inflammation. In this study, we demonstrate that sodium butyrate (SB), a short-chain fatty acid, induces a functional profile that diverges from the canonical pathways observed in classical innate immune training. The induction of an innate reprogrammed state in chicken macrophages by SB is strictly dependent on the cellular developmental stage, occurring only during the early stages of differentiation from chicken bone marrow-derived macrophages but not in fully differentiated cells. This suggests that SB primarily facilitates an innate immune reprogramming with a specific temporal window of sensitivity. Our results show that SB-reprogrammed chicken macrophages exhibit enhanced reactive oxygen species generation, altered cytokine expression, and an increased capacity to kill a diverse range of bacteria. Treatment with chemical inhibitors further demonstrated that these heightened antibacterial effects are directly attributed to increased reactive oxygen species production and autophagy. In summary, these findings indicate that SB induces functional outcomes distinct from classical trained immunity and can elicit innate immune memory through alternative regulatory axes. Our data suggest that distinct innate reprogramming states give rise to alternative activation programs and that innate immune memory exists along a spectrum of phenotypes rather than as a single, uniform state.
[This corrects the article DOI: 10.1039/D6RA00556J.].
Fire blight, caused by Erwinia amylovora, can cause yield losses of up to 95% in apple and pear orchards. Management commonly relies on antibiotic applications during bloom, which may promote antibiotic resistance. Here, bentonite nanoclay (BNT) composites loaded with clove, thymol, or oregano essential oils (EOs) were evaluated as antibiotic-free materials for fire blight control. In vitro assays showed that BNT-EO formulations inhibited E. amylovora growth, with the minimum inhibitory concentration (MIC) of oregano EO and BNT-oregano (O) estimated at 5 g·L-1. Field trials showed that BNT-EO formulations reduced pathogen populations and flower infection over two seasons. In 2023, BNT-O achieved 76% disease reduction, approaching streptomycin efficacy (81%), although efficacy declined in 2024. Elemental analysis confirmed significant enrichment of BNT-derived Al, Na, and Si on BNT-O-treated flowers 9 days postapplication (p < 0.001). These findings support EO-loaded nanoclays as promising delivery systems for sustainable fire blight management.
In this case, we present a patient with rapidly progressive unilateral destructive orbital and midface inflammation. Initial detection of Corynebacterium kroppenstedtii was followed by a consistent course of localised necrotizing small-vessel vasculitis, which was responsive to B-cell depletion. A 79-year-old woman presented with acute left periorbital pain, tearing and purulent discharge. The workup included serial microbiological cultures, fungal and mycobacterial testing, molecular diagnostics, CT/MRI/PET-CT imaging, multiple surgical biopsies with histopathology and special stains, ANCA testing, and multidisciplinary treatments (antimicrobials, glucocorticoids, methotrexate and rituximab). The clinical course, laboratory work, imaging studies, histological analysis and response to treatment were thoroughly reviewed. Initial conjunctival culture revealed C. kroppenstedtii and clindamycin and vancomycin was initiated. Despite escalating antimicrobials, the patient developed progressive inflammation with sinus destruction, cutaneous fistula and cheek abscess, requiring orbital exenteration. The biopsies revealed chronic inflammation with multinucleated giant cells, focal necrosis and small-vessel vasculitis. ANCA remained negative, and subsequent testing was negative. High-dose prednisolone improved the patient, prompting the discontinuation of methotrexate because of pancytopenia. Rituximab-induced remission allowed for steroid tapering and rituximab maintenance, which controlled the disease without complications. This case illustrates a rare, localised ANCA-negative necrotizing small-vessel vasculitis with destructive orbital involvement and initial detection of C. kroppenstedtii. Although the pathogenic significance of this finding remains uncertain, the case highlights the importance of comprehensive microbiological and immunological evaluation in atypical destructive inflammatory disease. Sustained disease control was achieved with immunosuppressive therapy including rituximab after persistent infection had been excluded.
The global health threat of antimicrobial resistance necessitates the development of novel therapeutic strategies. This study identifies the anticancer agent mitoxantrone as a potent antimicrobial against Klebsiella pneumoniae. Through passaging experiments, we demonstrate that resistance to mitoxantrone arises via mutations in the kstR2 gene, leading to the overexpression of the SmvA efflux pump. Transcriptomic analysis reveals that mitoxantrone treatment induces the bacterial SOS response, likely due to DNA damage. Critically, we found that mitoxantrone exhibits a synergistic effect with tigecycline against K. pneumoniae, both in vitro and in a murine wound infection model. Mechanistically, this synergy is driven by tigecycline-mediated enhancement of intracellular mitoxantrone accumulation. These findings reveal a potentially effective combination regimen to combat K. pneumoniae infections.IMPORTANCEThe rise of antibiotic-resistant Klebsiella pneumoniae demands new treatment options. This study uncovers that mitoxantrone, a drug currently approved for cancer therapy, is effective against K. pneumoniae. Furthermore, we discovered that combining mitoxantrone with the existing antibiotic tigecycline is synergistic against K. pneumoniae. We found that tigecycline enhances mitoxantrone accumulation inside bacterial cells. Overall, our work identified the tigecycline-mitoxantrone combination as a promising strategy to treat K. pneumoniae infections.
The increasing incidence of Candida glabrata complex infections in hospitalized patients and their association with high mortality rates prompted the determination of cryptic species among clinical isolates from a tertiary hospital and the evaluation of their susceptibility profiles to commonly prescribed antifungal agents. This study evaluated 80 C. glabrata isolates obtained from patients admitted to a Brazilian public tertiary hospital. The isolates were recovered from different clinical specimens, predominantly urine and blood, across various medical units. MALDI-TOF MS analysis revealed that all isolates were Candida glabrata sensu stricto. Minimum inhibitory concentrations (MICs), determined via broth microdilution and according to the European Committee on Antimicrobial Susceptibility Testing (EUCAST) guidelines, showed high susceptibility to amphotericin B (AmB), voriconazole (VRC), and echinocandins (ECNs) [caspofungin (CSF), anidulafungin (ANF), and micafungin (MCF)], as well as uniform susceptibility within the "I" category (susceptible, increased exposure) to fluconazole (FLC). Comparisons of resistance profiles revealed higher prevalences of resistance to AmB and VRC than to ECNs, both overall and in urine isolates, with a similar trend observed in blood isolates. Comparative analysis with reference EUCAST C. glabrata data highlighted higher localized MIC values for AmB, ANF, and MCF, lower MICs for FLC, and equivalent distributions for VRC. All recovered isolates were confirmed as Candida glabrata sensu stricto and exhibited different susceptibility profiles from reference EUCAST isolates. These findings establish a strong regional baseline, serving as a useful guide for monitoring therapeutic measures and supporting antimicrobial stewardship.
Streptococcus canis is a rare zoonotic pathogen reported to cause human disease, including skin and soft tissue infections, bloodstream infections, and infective endocarditis. The majority of cases have been linked to contact with dogs, with a paucity of cases related to cats. It is generally susceptible to penicillin and vancomycin but exhibits a high rate of resistance to erythromycin and an inducible resistance to clindamycin. A man in his sixties presented with fever, chills, and a diabetic foot ulcer that had been exposed to a cat. He was found to have S. canis primary bloodstream infection, which showed a typical susceptibility to beta-lactams and vancomycin, and resistance to clindamycin and macrolides. He was treated with ceftriaxone and had no recurrence at the 3-month follow-up. S. canis related to cat exposure is an extremely rare cause of human infection, in such cases, a beta-lactam antimicrobial should be initiated. In individuals with an allergy to beta-lactams, vancomycin should be used with avoidance of macrolides and clindamycin as empirical therapy due to the high resistance rate to these antimicrobials among S. canis.
Strychnos spinosa Lam. (monkey orange) is a drought-tolerant indigenous fruit tree distributed across sub-Saharan Africa, valued traditionally for nutrition and medicine but remaining underutilised in formal economies. This review synthesises literature on the fruit's physicochemical properties, phytonutrient profile, value-added products, and bio-accessibility to evaluate its potential as a functional food. A systematic literature search was conducted following PRISMA guidelines across database using different keyword strings. Of 387 initial records identified, 75 studies met inclusion criteria and were included in the qualitative synthesis. The pulp is characterised by high acidity (pH 2.5-4.0), soluble solids (12-22 °Brix), and pulp yield (30-60%). Nutritionally, it is rich in carbohydrates, dietary fibre, and minerals including potassium, magnesium, calcium, iron, and zinc. Phytochemical profiling revealed bioactive compounds such as chlorogenic acid, caffeic acid, rutin, and carotenoids, underpinning antioxidant, antimicrobial, and anti-inflammatory activities. Value-added products including juices, jams, fermented beverages, and dried rolls have demonstrated feasibility. However, a critical knowledge gap persists: while compositional analyses confirm high phytonutrient content, their bio-accessibility remains poorly characterised. Factors including carotenoid lipophilicity, phenolic compound stability during digestion, and processing impacts on nutrient release have not been systematically quantified. Without integrating standardised bio-accessibility studies, the physiological relevance of S. spinosa's phytonutrients cannot be validated, and commercialisation as a functional food remains speculative. Fermentation emerges as a promising traditional and scalable method to enhance nutrient bioavailability through phenolic modification, cell wall disruption, and reduction of anti-nutritional factors. This review recommends prioritising bio-accessibility investigations alongside fermentation optimisation to transform S. spinosa into a validated, climate-smart functional food for sub-Saharan Africa, supporting evidence-based product development, nutrition security, and rural economic opportunities.
Idiopathic Granulomatous Mastitis (IGM) is a rare, chronic inflammatory breast condition with uncertain etiology and challenging management. Standard treatments often involve corticosteroids or surgery, both associated with significant relapse and adverse effects. Recent interest has emerged in using antibiotics such as Trimethoprim-Sulfamethoxazole (TMP-SMX) due to potential antimicrobial and anti-inflammatory benefits, though evidence remains limited. This retrospective cohort study evaluated the effectiveness and safety of TMP-SMX compared to conventional therapies in female patients diagnosed with histologically confirmed IGM. Patients were categorized into the TMP-SMX and control groups. Primary outcomes included treatment response and recurrence rates; secondary outcomes assessed changes in lesion size, clinical severity, and adverse events. Statistical analyzes included Chi-square, Fisher's exact test, and odds ratios (ORs) with 95% confidence intervals. Complete clinical remission was achieved in 70.8% of TMP-SMX (N = 24) patients versus 51.2% in the control (N = 43) group (p = 0.12). Recurrence rates were comparable between groups (37.5% in the TMP-SMX group vs. 27.9% in the control group, p = 0.51). TMP-SMX patients showed trends toward reduced lesion size and milder disease, although differences were not statistically significant. Adverse events were less frequent in the TMP-SMX group, except for transient fatigue (p = 0.04). This study provides evidence that TMP-SMX is a safe and clinically effective alternative to corticosteroid-based regimens for managing idiopathic IGM. TMP-SMX demonstrated comparable efficacy, including favorable trends in lesion regression and disease severity improvement, with notably fewer systemic adverse events.
The dewberry, or Rubus caesius L. (Rosaceae), has long been used in traditional medicine, although scientific research has primarily focused on its roots and aerial portions. Notable biological activity associated with a rich phytochemical profile has been reported in these investigations. In contrast, limited information is available regarding the biological potential and chemical composition of R. caesius roots collected from Turkmenistan, Central Asia. The present study aimed to investigate, for the first time, the biological potential of R. caesius root extracts prepared as an aqueous infusion (a traditional, food-relevant preparation) and an ethanol extract (a polyphenol-enriching extraction), and to relate differences in bioactivity to UHPLC-QqQ-MS/MS-based phytochemical composition. The extracts were screened using complementary antioxidant assays (radical-scavenging and reducing power), a multi-target panel of enzyme inhibitory activities relevant to metabolic and cosmetic/neurobiological pathways (including α-amylase, α-glucosidase, acetyl-/butyrylcholinesterase, and tyrosinase), and antimicrobial/antifungal tests against representative bacterial and fungal strains. UHPLC-QqQ-MS/MS profiling revealed a root metabolome dominated by phenolic acids, flavonoids, and anthocyanin-related constituents, with clear qualitative and quantitative differences between infusion and ethanol extracts. In agreement with the chemical findings, both extracts expressed pronounced antioxidant capacity, while enzyme inhibition was extract-dependent, indicating that solvent polarity influenced the recovery of constituents responsible for carbohydrate-hydrolyzing enzyme and cholinesterase/tyrosinase inhibition. The antimicrobial and antifungal screenings demonstrated selective growth inhibition that varied by microorganism and extraction type. The integrated chemical-biological approach highlights R. caesius roots from Turkmenistan as a promising source of multifunctional natural products and provides baseline data to support their future use in nutraceutical, functional food, or phytopharmaceutical development.
Persistent intestinal carbapenemase-producing Enterobacterales (CPE) carriage challenges infection prevention and antimicrobial stewardship. We evaluated fecal microbiota transplantation (FMT) for decolonization and response-associated recipient microbiome features. This prospective cohort study assessed intestinal CPE clearance in 131 adult carriers (68 receiving FMT; 63 under observation). Responders achieved clearance, defined as three consecutive negative rectal surveillance cultures for CPE, obtained at 3-day intervals within 1 month after FMT. Following least absolute shrinkage and selection operator (LASSO)-based covariate selection, Cox proportional hazards models estimated associations between FMT and time to successful decolonization within prespecified 0-30-day and 0-90-day windows. A longitudinal metagenomic subcohort of 21 FMT recipients (102 fecal samples) underwent shotgun sequencing to characterize taxonomic composition, microbial network organization, functional pathways, antimicrobial resistance genes (ARGs), and donor engraftment. After LASSO-based covariate selection, FMT was associated with faster CPE clearance than observation at 1 month (hazard ratio, 4.02; 95% confidence interval, 1.84-8.79), with effects sustained at 3 months. Responders showed relatively preserved baseline microbial network organization and enrichment of taxa annotated with arginine-related pathway features, suggesting metabolic relevance to microbial niche competition. After FMT, responders had greater engraftment of donor-associated taxa and donor-like ecological reassembly, with reduced Klebsiella pneumoniae dominance and ARG abundance. FMT may accelerate intestinal decolonization in CPE carriers; response variability was associated with recipient gut-microbiome ecology, suggesting microbiome-guided patient selection could optimize microbiota-based strategies.