Search Results (2,621)

Background: Nosocomial infections caused by multidrug-resistant microorganisms are a significant public health concern. Antimicrobial resistance (AMR) is closely linked to the excessive and indiscriminate use of antibiotics, which creates selective pressure and promotes the emergence of resistant pathogens. Objectives: This study evaluates the synergistic potential of intragenic antimicrobial peptides (IAPs) combined with plant alkaloids against susceptible and multidrug-resistant human pathogenic bacteria, assessing antimicrobial activity, biofilm inhibition, and hemocompatibility. Methods: The tested molecules included berberine, tomatidine, sinomenine, and the IAPs Hs02 and Gr01. Minimum inhibitory concentration (MIC) and minimum microbicidal concentration (MMC) assays were performed against both ATCC (E. coli ATCC 25922 and S. aureus ATCC 25923) and clinical strains (E. coli KPC+ HRAN 1812446 and S. aureus MDR LACEN 3730529). Synergistic interactions were evaluated by checkerboard assay, followed by biofilm inhibition and hemolysis assays using human red blood cells. Results: Berberine exhibited a MIC of 1024 µM when tested individually, while tomatidine and sinomenine showed no significant activity. As expected, the IAPs showed strong antimicrobial properties at 8 µM (Hs02) and 4 µM (Gr01). When tested in synergy, alkaloids and IAPs reduced the MIC by up to 128-fold. The combination of IAPs and alkaloids reduced the biofilm biomass of S. aureus and E. coli by 50%, by the crystal violet assay (p < 0.05). Notably, sinomenine had not previously been reported to have antimicrobial activity. Conclusions: These results highlight the importance of further exploring combinations of natural and synthetic bioactive molecules as promising antimicrobial candidates. This approach may help to extend the useful life of conventional antibiotics. However, further studies are needed to assess safety, cytotoxicity, genotoxicity, inflammation, and in vivo effects. Full article

Background/Objectives: Following the introduction of the 13-valent pneumococcal conjugate vaccine (PCV13) in Oman, this study aimed to characterize the genomic epidemiology, serotype distribution, and antimicrobial resistance (AMR) of Streptococcus pneumoniae causing invasive pneumococcal disease (IPD). Methods: All IPD isolates collected through national laboratory-based surveillance between 2018 and 2021 were analyzed using Whole-Genome Sequencing (WGS). Bioinformatics tools determined serotypes, multilocus sequence types (MLSTs), and Global Pneumococcal Sequence Clusters (GPSCs). Clinical correlates and predictors of mortality were assessed via multivariate logistic regression. Results: A total of 129 IPD isolates were included. Serotype 3 (11.6%) was the most prevalent, followed by 23B and 9N (10.8% each), and 8 (8.5%). PCV13 serotypes accounted for only 26.4% of isolates, while PCV20 coverage reached 59.7%. Significant clonal diversity was observed, with GPSC12 (Serotype 3) and GPSC699 (Serotype 9N/13) being prominent lineages. Multidrug resistance (MDR) was identified in 36.4% of isolates, primarily driven by GPSC6 and GPSC699. The case fatality rate was 23.0%. Advanced age (≥65 years) and clinical presentation with bacteremia were significant independent predictors of death, whereas bacterial genotype and AMR status were not. Conclusions: The findings demonstrate significant serotype replacement in Oman after the introduction of PCV13. The high prevalence of non-vaccine serotypes and emerging MDR clones justifies the transition to higher-valency vaccines like PCV20. Sustained genomic surveillance remains essential to monitor the evolving landscape of invasive pneumococcal lineages. Full article

Background/Objectives: Klebsiella (K.) pneumoniae is a significant pathogen in both humans and animals. However, data on its occurrence in animals in Germany remain limited. This study aimed to investigate the antimicrobial resistance (AMR) phenotypes, AMR genes, and virulence traits of animal-derived K. pneumoniae isolates from Germany. Methods: A total of 59 K. pneumoniae isolates obtained in 2023 from dogs, cats, horses, cattle, and chickens across 11 German federal states were analysed. Phenotypic antimicrobial susceptibility testing (AST) was performed, and whole-genome sequencing (WGS) was used for genomic characterisation, including detection of AMR genes, virulence-associated genes, sequence types (STs), and plasmid replicons. Results: Most isolates (78%) were susceptible to all tested antibiotics, while three isolates were classified as multidrug-resistant (MDR). Resistance was most frequently observed for piperacillin (n = 8) and trimethoprim/sulfamethoxazole (n = 4). Carbapenem resistance was detected in two isolates (one from a dog and one from a cat), and phenotypic colistin resistance in one dog isolate. WGS identified 96 AMR genes across isolates, with 20–42 AMR determinants per isolate, conferring resistance to β-lactams, aminoglycosides, fluoroquinolones, sulfonamides, tetracyclines, trimethoprim, and fosfomycin. Ten extended-spectrum β-lactamase (ESBL)-producing isolates carried genes including bla_CTX-M-15, bla_SHV-2, bla_SHV-27, bla_SHV-42, bla_SHV-106, and bla_TEM-158. Although fosA was detected in all isolates, only three exhibited phenotypic resistance to fosfomycin. A total of 52 STs were identified, including high-risk clones. One hypervirulent isolate (ST60) carrying hypervirulence-associated genes rmpA and iroB was detected. Plasmid replicons were present in 70% of isolates, while plasmid-associated AMR genes were identified in nine isolates. Conclusions: This study demonstrates the genomic diversity of K. pneumoniae identified in companion animals and highlights the presence of AMR and virulence determinants relevant to a One Health context. Full article

Background/Objectives: Environmental dissemination of antimicrobial resistance (AMR) is increasingly driven by wastewater-impacted aquatic systems, yet the key factors controlling multidrug-resistant (MDR) bacterial distribution remain unclear. This study evaluated environmental factors associated with MDR bacteria in the urban Danube River (Novi Sad, Serbia) using a machine learning framework. Methods: Surface-water and wastewater samples were collected during summer and autumn 2024. Bacterial isolates were obtained through membrane filtration onto chromogenic media and identified using MALDI-TOF MS. Physicochemical parameters (including COD, BOD₅, turbidity, pH, and temperature) were used as predictors in seven machine learning models (ANN, RF, SVM, XGB, MARS, TREE, NB). Model performance was assessed using AUC, accuracy, and error metrics. Results: Wastewater samples showed higher bacterial abundance and taxonomic richness than river surface-water samples, with frequent recovery of Klebsiella pneumoniae, Escherichia coli, Aeromonas veronii, and Pseudomonas spp. Tree-based models (RF, XGB) performed best. Organic pollution indicators, turbidity, pH, and water temperature were the most prominent factors. Conclusions: Wastewater-related pollution gradients, reflected by organic load parameters, turbidity, pH, and water temperature, were associated with the occurrence of selected bacterial taxa recovered on selective media. These associations were more pronounced in wastewater samples, while river surface-water samples showed lower abundance and taxonomic richness but still contained selected environmental and opportunistic taxa. Because antimicrobial susceptibility testing and molecular confirmation of resistance determinants were not performed, the findings should be interpreted as culture-based and exploratory. Machine learning approaches may support environmental screening and hypothesis generation for AMR-oriented surveillance, but future studies should include standardized phenotypic and molecular confirmation. Full article

Background/Objectives: The emergence of colistin resistance mediated by the mcr-1 gene in Salmonella enterica poses a significant public health concern. In 2022, mcr-1 was identified for the first time in Salmonella isolates recovered from chicken meat in Saudi Arabia, prompting the need for further genomic investigation. Methods: Four Salmonella enterica isolates—two serovar Minnesota and two serovar Infantis—underwent antimicrobial susceptibility testing (AST), whole-genome sequencing (WGS), and bioinformatics analyses including antimicrobial resistance genes (ARGs), virulence factors, plasmid replicons, and mcr-1 locations. Results: All isolates exhibited resistance to colistin, polymyxin B, and multiple first-line antibiotics. All four isolates were classified as multidrug-resistant (MDR). The mcr-1 gene was plasmid-borne in all isolates, located on IncI1_1_Alpha plasmids in S. Minnesota and IncFIB(K)_1_Kpn3 plasmids in S. Infantis. Additional antimicrobial resistance genes (ARGs) were detected, including bla_CTX-M-65, qnrB5, ermB, and aminoglycoside-modifying enzymes, alongside multiple efflux pump genes. Virulence gene profiles showed minor differences between serovars, including the presence of the cdtB toxin gene in S. Infantis isolates. Phylogenetic analysis indicated that the isolates clustered within a distinct clade, suggesting a potential common local source or clonal expansion. Conclusions: This study provides the first detailed genomic insight into mcr-1-positive Salmonella isolates from food in Saudi Arabia. The co-existence of resistance and virulence determinants, together with mobile plasmids carrying the mcr-1 gene, emphasizes the risk of dissemination through the food chain. These findings highlight the urgent need for integrated genomic surveillance and strengthened antimicrobial stewardship within a One Health framework. Full article

Growing resistance to carbapenem antibiotics is a major public health problem as these antibiotics are considered the last line of therapy for infections caused by multidrug-resistant (MDR) Gram-negative bacteria. The rapid emergence and dissemination of carbapenem-resistant bacterial strains are mainly due to horizontal gene transfer (HGT) within or between bacterial cells via the mobilome. The aim of this article is to discuss the role of mobile genetic elements (MGEs) that capture and disseminate resistance determinants of carbapenem antibiotics, as a comprehensive review integrating the combined role of plasmids, transposons and integrons. It attempts to systematically fill the gap by investigating the role of these MGEs in the acquisition, mobilization and dissemination of genes encoding carbapenemases across clinically important bacteria. Various types of plasmids such as IncF and IncH in Klebsiella pneumoniae, IncL/M in Enterobacter cloacae, IncX3 in Escherichia coli and IncA/C₂ in Salmonella enterica carry important genes encoding carbapenemases. The rapid distribution of transposons among bacterial species is one of the main contributing factors in the dissemination of carbapenem-resistant isolates. Transposons including Tn4401 carrying bla_KPC in K. pneumoniae and Tn1721 carrying bla_KPC in E. coli; Tn2006, Tn2007, Tn2008 and Tn2009 carrying bla_OXA-23 in Acinetobacter baumannii; Tn1696 carrying bla_IMP-4 in Pseudomonas aeruginosa; Tn125 carrying bla_NDM in E. coli; and Tn6306 carrying bla_IMI in Raoultella ornithinolytica encode different types of carbapenemases. Integrons mainly belonging to class 1 capture resistance determinants for metallo-carbapenemases such as NDM-, VIM-, SIM- and IMP-type enzymes in P. aeruginosa, A. baumannii, K. pneumoniae and E. coli and can promote the transcription and expression of these determinants. These findings are useful for understanding the genetics of carbapenem resistance and additional knowledge on MGEs may provide avenues for screening of resistance to these antibiotics in clinical settings. Full article

The diagnosis of enteric fever has become difficult due to the nonspecific and overlapping clinical syndrome of typhoid and paratyphoid infections with other febrile illnesses. Moreover, the rapid emergence of fluoroquinolone-resistant typhoidal Salmonella and the lack of robust diagnostic methods highlight the urgent need for highly sensitive molecular techniques. Here, we evaluated the performance of a rapid, reliable, and cost-effective molecular diagnostic approach for detecting Salmonella Typhi, including the globally dominant haplotype H58 lineage (H58), and Salmonella Paratyphi A. An in-house-built conventional polymerase chain reaction (PCR) was performed on a collection of blood-culture-positive strains, and the sensitivity and specificity were compared with those of the standard blood culture results. H58 and non-H58 Typhi lineages with distinct resistance patterns were confirmed from the previously reported sequencing data. Our PCR result showed that target genes SSPA2308, STY2513, and STY0307 demonstrated 100% sensitivity and specificity for Salmonella Paratyphi A, Salmonella Typhi, and H58 Salmonella Typhi, respectively. The PCR assay reliably detected bacterial DNA at 5.2 × 10⁴ colony-forming units (CFUs), with consistent amplification observed up to 10⁻¹ dilution. This single-nucleotide polymorphism (SNP)-based diagnostic approach has added a new dimension to designing unique markers for multidrug-resistant (MDR)-associated H58 lineage detection and has the potential to inform local treatment algorithms. Full article

Salmonella enterica serovar Enteritidis is a leading cause of foodborne zoonoses worldwide. The rapid emergence of multidrug-resistant (MDR) strains has compromised traditional antimicrobial therapies, necessitating the development of biosafe alternatives such as bacteriophages. This study aimed to isolate and comprehensively characterize novel lytic bacteriophages targeting multidrug-resistant Salmonella enterica subspecies enterica serovar Enteritidis isolates from Lebanon. In this study, four novel Salmonella phages, EDA02, EDA03, EDA05, and EDA06, were isolated from wastewater and poultry effluents in Lebanon. The isolates were characterized using host range profiling, one-step growth kinetics, and physicochemical stability assays. Comprehensive whole-genome sequencing (WGS) and phylogenetic analyses were performed to assess their genomic safety and taxonomic placement. Phages EDA03 and EDA06 exhibited the broadest intra-serovar lytic activity within the tested panel, infecting up to 72% and 67% of the MDR isolates, respectively. One-step growth analysis revealed latent periods of 30–40 min, with burst sizes ranging from 6.0 to 150 phages/infected cell. All four phages demonstrated robust stability across pH 4.7–10.3 and temperatures from 4 °C to 50 °C. WGS revealed genome sizes ranging from 42.3 kb to 108.8 kb, with no identified genes associated with lysogeny, virulence, or antimicrobial resistance. Phylogenomic analysis assigned all isolates to the family Straboviridae, with <95% intergenomic similarity to their closest RefSeq relatives, supporting their classification as novel species. The isolated phages demonstrate substantial lytic activity and environmental resilience under the tested conditions. Their complementary lytic profiles, environmental resilience, and genomic safety support their further evaluation as biocontrol candidates. This study represents the first genomic and phenotypic characterization of anti-Salmonella Enteritidis phages from Lebanon. These findings support the development of phage-based interventions for food safety and antimicrobial resistance mitigation in resource-limited settings. Full article

Coagulase-negative staphylococci (CoNS) and mammaliicocci (MA) are common in food-derived samples and may act as antimicrobial resistance (AMR) reservoirs. A previous study reported a high S. aureus prevalence in poultry meat. The objective of this study was to characterize the species diversity, antimicrobial resistance, and biofilm-forming capacity of CoNS/MA from the same food samples. Species identification, antimicrobial susceptibility testing, resistance gene detection, molecular typing, and biofilm formation assays were performed. One hundred and forty-eight non-repetitive CoNS/MA isolates were detected in 85% of samples, and 14 species were identified. The most prevalent species were S. epidermidis (18.2%), S. simulans (12.8%), S. saprophyticus (12.2%), S. warneri (11.5%), and M. lentus (10.1%). Most samples harbored one or two different species, although some showed higher diversity. Although 27.0% of isolates were pan-susceptible, 22.3% were multidrug-resistant (MDR), significantly associated with M. lentus and S. epidermidis. Methicillin resistance was found in 10 isolates, mainly in S. epidermidis (lineages ST9, ST59, ST88 and ST640). Biofilm formation was observed in 24.3% of isolates (some of them MDR) and was significantly associated with S. pasteuri and S. xylosus and with samples from supermarkets. No methicillin-resistant isolates were biofilm producers. These findings highlight the diversity of CoNS/MA in poultry meat and their role as AMR reservoirs and persistence factors, emphasizing their relevance in food safety. Full article

Background/Objectives: Biofilms are a form of defense that enables bacteria to withstand antibiotic pressure and demonstrate antibiotic resistance. It is crucial to develop anti-biofilm strategies in order to combat chronic and persistent multidrug-resistant (MDR) infections. Methods: In this study, we developed 3D biofilms of single-, dual-, and poly-species MDR ESKAPE components, including the pathogens P. aeruginosa S. aureus and K. pneumoniae, in CF Mu3Gel. We evaluated the efficacy of using a phage, a di-hetero phage cocktail or a poly-hetero phage cocktail in combination with ciprofloxacin to eliminate mature biofilm biomass after 72 h or one week in a single treatment. Results: The phage components mostly exhibited synergistic behavior when combined with ciprofloxacin and with each other in di- and poly-hetero-cocktails. The reduction in 72-h dual- and poly-species biofilms was one log higher than that of one-week biofilms treated with the phage–antibiotic combination. The greatest reductions were observed in the 72-h single-species biofilm with combination therapy, at 1.4–3.0 log. Reductions of 2.16 and 1.6 log were observed in the dual-species P. aeruginosa and S. aureus biofilm and the poly-species biofilm, respectively. Conclusions: This study examined how a single application of phages or phage cocktails, either alone or in combination with ciprofloxacin, impacted established biofilm models, and how this affected the proportion of microcolonies of different species within each model. These insights will facilitate the development of strategies for multiple follow-up treatments, as well as the reordering of phages, phage cocktails, and combinations with antibiotics, to improve outcomes. The 3D biofilm models developed here could be used to screen phages or phage cocktails either on their own or alongside other therapies. This would facilitate the application of in vitro findings to real physiological settings. Full article

Background: Mobile health has been increasingly integrated into tuberculosis care to support patient education, communication, and treatment engagement. However, evidence remains limited regarding whether positive engagement with mHealth is associated with knowledge, attitudes, and practices among patients with multidrug-resistant tuberculosis. This study aimed to examine the association between positive mHealth engagement and knowledge, attitude, practice, and total KAP among patients with multidrug-resistant tuberculosis, and to evaluate the psychometric properties of the engagement score used as the primary exposure variable. Methods: A cross-sectional study was conducted among patients with multidrug-resistant tuberculosis. A positive mHealth engagement score was constructed from 12 mHealth-related items after harmonizing item directionality so that higher scores indicated more favorable engagement. The 12 items reflected five behavioural domains: intensity of use, ease and acceptability of use, functional engagement (communication with providers, access to health information, and perceived benefit for disease self-management), continuity of use, and barriers to sustained engagement. The composite score was computed as the mean of the 12 standardised items, with higher values indicating more positive engagement. Internal consistency was assessed using Cronbach’s alpha and corrected item–total correlations, and structural validity was explored using principal component analysis. Adjusted linear regression models were used to examine associations between the engagement score and Knowledge, Attitude, Practice, and total KAP scores, controlling for age, sex, and occupation. Sensitivity analyses were performed after excluding a poorly performing item, and tertile analyses were used to assess dose–response patterns. Results: The positive mHealth engagement score showed good internal consistency, with a Cronbach’s alpha of 0.852. One item demonstrated poor psychometric performance, and Cronbach’s alpha increased to 0.864 after its exclusion. The data were suitable for dimensionality assessment, with a Kaiser–Meyer–Olkin value of 0.870 and a significant Bartlett’s test. Principal component analysis identified a dominant first component explaining 43.29% of the total variance. Using the refined score, higher positive mHealth engagement was significantly associated with higher Knowledge scores (β = 2.06; 95% CI: 1.28–2.85; p < 0.001), higher Attitude scores (β = 4.68; 95% CI: 3.30–6.06; p < 0.001), and higher total KAP scores (β = 6.68; 95% CI: 4.62–8.74; p < 0.001), whereas no significant association was observed for the Practice score (β = −0.07; 95% CI: −0.63 to 0.49; p = 0.804). In tertile analyses, Knowledge, Attitude, and total KAP scores increased significantly across engagement levels, while Practice scores did not. Conclusions: Positive mHealth engagement was associated with better knowledge, attitudes, and overall KAP among patients with multidrug-resistant tuberculosis, but not with practice. These findings are associative; the cross-sectional design does not permit causal conclusions. The engagement score demonstrated good reliability and acceptable structural validity and may be a useful summary measure for evaluating patient interaction with mHealth interventions in tuberculosis care. Integrated strategies combining mHealth with clinical follow-up, adherence counseling, and structural support may be needed to translate informational and attitudinal gains into practice change. Full article

Background: Acinetobacter baumannii (A. baumannii) is a critical-priority pathogen of major concern in healthcare settings. Colistin remains a last-resort antibiotic for multidrug-resistant (MDR) A. baumannii infections; however, resistance is increasingly reported worldwide yet remains understudied in Bahrain. Reliable detection methods and understanding clonal dissemination are essential for infection control. Objectives: This study aimed to (1) determine the rate of colistin resistance in 102 clinical A. baumannii isolates from Bahrain, (2) evaluate the diagnostic performance of the colistin agar test (CAT) and E-test against broth microdilution (BMD method), and (3) assess clonal relationships using BOX-PCR fingerprinting. Methods: 102 clinical isolates from multiple hospitals in Bahrain underwent susceptibility testing via the BMD method, CAT, and E-test; screening for mcr-1 to mcr-5 genes; and BOX-PCR DNA fingerprinting. Results: Colistin resistance was detected in 14.7% of isolates by BMD method, higher than regional and global averages. All resistant isolates were mcr-negative, suggesting chromosomally mediated resistance. CAT showed 86.7% sensitivity, 98.8% specificity, and a 13.3% very major error rate. The E-test failed to detect resistant isolates (very major error 100%). BOX-PCR revealed predominant clonal relatedness with intra- and inter-hospital spread. Conclusions: Colistin resistance in A. baumannii from Bahrain exceeds regional and global levels, likely driven by chromosomal mechanisms under selective pressure. The BMD method remains the gold standard for colistin testing, while CAT may serve as a screening tool requiring confirmation. Strengthened stewardship and infection control measures are vital to contain dissemination. Full article

Background/objectives: Infections caused by multidrug-resistant (MDR) bacteria are becoming increasingly difficult to treat with recommended antimicrobials. Considering the critical and growing challenge of antimicrobial resistance (AMR), this study aims to evaluate the antimicrobial susceptibility patterns of Escherichia coli clinical isolates from dogs in Grenada. This research project consists of two distinct studies: a retrospective analysis of AMR in canine E. coli isolates collected between 2010 and 2020, and a cross-sectional study characterizing the genotypic AMR profiles of E. coli isolates obtained between April and June 2023. Methods: A retrospective analysis of antibacterial sensitivity test (ABST) reports from canine clinical samples submitted to the Small Animal Clinic at St. George’s University (SGU), St. George’s, Grenada, between 2010 and 2020 revealed a notable prevalence of AMR among canine E. coli isolates. To further investigate the underlying mechanisms of this resistance, the study analyzed canine E. coli isolates that exhibited phenotypic resistance in ABST assays. These isolates were subsequently screened for AMR-associated genes using polymerase chain reaction (PCR) and next-generation sequencing (NGS). Results: The retrospective study identified 153 canine clinical isolates positive for E. coli. The antimicrobial drugs, imipenem, cefotaxime and ciprofloxacin were found to be highly effective against these isolates. However, a gradual increase in AMR was observed for amoxicillin–clavulanic acid (34.88%), ampicillin–sulbactam (17.31%), cephalexin (43.08%), cefpodoxime (22.31%), cephalothin (68.42%), and doxycycline (37.04%). In the prospective study, PCR analysis of resistant canine E. coli isolates detected the tetA (577 bp) and blaTEM (686 bp) genes. These AMR determinants were further confirmed through analysis of NGS reads and assembled contigs. Additionally, NGS-based predictions identified genes associated with resistance to aminoglycosides and potentiated sulfonamides. Conclusions: This study demonstrates that E. coli from dogs in Grenada exhibits resistance to tetracycline and several β-lactam antimicrobials. These findings underscore the need for rational antimicrobial stewardship and continuous AMR surveillance in small animal practice within the region. Full article

Antibiotic resistance remains a major public health concern, particularly in regions with high rates of hospital- and community-acquired infections. This study aimed to quantify multidrug-resistant (MDR) bacterial strains in northern Mexico and to identify the most prevalent resistance phenotypes, the antibiotic classes with the highest resistance and susceptibility rates, the predominant MDR species, and the specimen types yielding the greatest number of isolates. Clinically relevant strains were collected from patients with confirmed infections. Microorganism identification and antimicrobial susceptibility test-ing were performed using the Vitek 2 Compact system (bioMérieux), and the results were analyzed descriptively. Of the 1544 strains analyzed, 761 (49.29%) exhibited multidrug resistance. Escherichia coli was the most frequently isolated MDR species, followed by Pseudomonas aeruginosa. Acinetobacter baumannii showed the highest resistance rate, with 95.55% of its strains classified as MDR, whereas P. aeruginosa had the lowest MDR proportion at 30.73%. These findings underscore the urgent need for rational antibiotic use and the development of new therapeutic agents, particularly those targeting Gram-negative bacilli, to mitigate the growing threat of antimicrobial resistance in this region. Full article

Background/Objectives: Escherichia coli (E. coli) spans commensal, intestinal pathogenic, and extraintestinal pathogenic lineages distributed across human, animal, and environmental reservoirs, yet the extent to which virulence architectures are shared across these compartments remains incompletely understood. Using a One Health framework, we profiled putative virulence determinants in pooled multidrug-resistant (MDR) E. coli source groups representing human, animal, and environmental sectors. Methods: Virulence genes were predicted with VirulenceFinder, and presence–absence profiles were integrated to define functional composition, sector overlap, source-group distribution breadth, and pathotype-associated signatures. Predicted pathogenic potential was assessed with PathogenFinder and compared with pathogenic family richness. Results: Overall, 114 putative virulence genes were detected, with adhesion/colonization functions dominating the virulome (33/114), followed by toxin-associated genes (12/114). A conserved core of 50 virulence genes was shared across all three sectors, including determinants linked to serum resistance (iss, ompT, traT), adhesion (csgA, fimH), stress adaptation (terC), and iron acquisition (sitA, iutA, fyuA). ExPEC-associated determinants were most numerous in environmental source groups (n = 52), whereas diarrheagenic E. coli markers were most frequent in animal-associated groups (n = 42). LEE-associated effectors were infrequent and largely absent from human source groups. Despite ecological differences in virulence composition, pathogenicity scores remained consistently high across sectors (0.83–0.92) and showed no significant association with pathogenic family richness (Spearman’s ρ = 0.197, p = 0.392). Conclusions: Within the limits of pooled source-group analysis, these findings suggest that MDR E. coli across One Health compartments shares a broadly distributed, ExPEC-associated virulence repertoire overlaid with sector-specific pathotype signals, underscoring the value of integrated genomic surveillance while highlighting the need for isolate-resolved analysis. Full article