Tongue squamous cell carcinoma (TSCC) is clinically heterogeneous, and patients with a similar TNM stage can experience markedly different outcomes. We systematically reviewed omics-driven studies to identify prognostic TSCC biomarkers. Although fundamentally prognostic, we discussed their theoretical translational relevance regarding future clinical decisions—such as treatment stratification or surveillance intensity—while strictly framing them as preliminary, hypothesis-generating targets. PubMed, Scopus, Web of Science, and Cochrane were searched for original human studies published between 2014 and 2024 using high-throughput genomic or transcriptomic profiling. Study selection followed referred Reporting Items for Systematic Reviews and Meta-Analyses (PRISMA), data were extracted with a structured workbook, and risk of bias was assessed using QUIPS and PROBAST, with reporting completeness appraised using REMARK. Seventeen studies were included, identifying 85 distinct biomarkers. Across biomarkers supported by multivariable overall survival analyses, higher-risk associations were reported for NELL2, PDE4D, CTTN, HBEGF, and CA9, whereas lower-risk associations were reported for AC139530.1, LINC01711, CCDC96, CYP2J2, and SPAG16. Recurrent biological themes included IL-17 signaling, ECM-receptor interaction, and focal adhesion. CA9 was the only biomarker reported in more than one included study, supporting its prioritization for validation. Although the evidence remains heterogeneous and largely hypothesis-generating, these markers may support the future validation of response-oriented therapeutic stratification in TSCC.

Conflicting data exist regarding the effect of intradetrusor BoNT/A on the incidence of urinary tract infections (UTIs) in patients with neurogenic detrusor overactivity (NDO), contrary to the increase in UTIs noted in patients with idiopathic OAB. Associations between UTIs, chronic inflammation, and bladder overactivity are acknowledged, albeit not fully understood. Chronic bladder inflammation is common in both NDO and OAB patients, and both animal and human studies suggest a beneficial effect of BoNT/A on both urinary and systemic levels of inflammatory markers. To explore whether intradetrusor BoNT/A injections affect the background for the incidence of UTIs in humans, we investigated in parallel the effect of intradetrusor BoNT/A on the incidence of UTIs and on the urine mRNA levels of urinary pathogen-detecting Toll-like receptors TLR2, TLR4, and TLR5 and of factors acting as intermediates of immune response and promoters of inflammatory reactions (IL1β, IL6, TNFα, and PGE2). For this purpose, we recruited 22 patients with NDO-associated incontinence who received at least one bladder BoNT/A injection. Urine specimens for the study of UTIs were obtained before the procedure and at routine urodynamic follow-ups at 4-6 weeks, 6 and 12 months post-BoNT/A, and at clinical relapse, while urine specimens for the study of biomarkers were collected at the time of BoNT/A injection and at the abovementioned follow-ups thereafter. Urine specimens from 10 adult healthy volunteers with no OAB symptoms served as the control group in the biomarker study. The genes of interest in the urine were studied by RNA isolation, reverse transcription, and real-time PCR. The urine mRNAs of all biomarkers tested appeared to be upregulated in the patients' samples compared with the controls, albeit only TLR2 and TLR5 mRNA increases were statistically significant. A progressive downregulation of TLR2, TLR5, IL1β, and IL6 urine mRNAs was noted at one and six months post-BoNT/A. TNFα and PGE2 mRNAs showed a transient increase at one month post-BoNT/A followed by a dramatic drop at the six months' follow-up. A similar trend for progressive decline was also noticed in the prevalence of both positive urine cultures and symptomatic UTIs in the same timepoints and additionally at 12 months post-treatment in patients who still benefited from the BoNT/A treatment. Upon clinical relapse, the mRNA levels of PGE2, IL1β, and IL6 increased in parallel with an increase in the prevalence of UTIs, while the levels of TLRs and TNF-α did not follow the same trend. In summary, intradetrusor BoNT/A injections achieved significant decreases in the urine mRNA levels of pathogen-detecting TLRs, immune response, and inflammation mediator cytokines and PGE2 in our cohort of patients with NDO-associated incontinence. In parallel, decreases were noted in both the incidence of symptomatic UTIs and rates of positive urine cultures. At the time of clinical relapse, the markers of inflammation and immune response, but not TLRs, were upregulated in parallel with the increased incidence of UTIs, suggesting that the studied genes PGE2, IL1β, and IL6 could be further explored as potential biomarkers for inflammation/immune response and UTIs in the neurogenic population.

Uterine leiomyosarcoma (uLMS) is a rare but highly aggressive mesenchymal malig-nancy of smooth muscle origin that represents a significant therapeutic challenge due to its poor prognosis and limited treatment options. Despite advances in molecular charac-terization, diagnosis remains difficult, and systemic therapies have shown limited success, contributing to a high recurrence rate and poor survival. Recent proteomic investigations have provided new insights into uLMS biology by exploring the global protein expres-sion patterns that define malignant transformation and progression. Using high-resolution mass spectrometry (MS)-based techniques, quantitative proteomics, and integrated multi-omics approaches, researchers have begun to identify dysregulated proteins, signaling pathways, and post-translational modifications (PTMs) linked to tumor metabolism, extracellular matrix (ECM) remodeling, cell-cycle regulation, and chemo-resistance. These studies have also uncovered candidate biomarkers that may improve the discrimination of uLMS from benign leiomyoma and have proposed novel therapeutic targets associated with metabolic reprogramming, kinase activation and tumor micro-environment (TME) modulation. This review summarizes recent advancements in uLMS proteomics, discusses their methodological underpinnings, highlights key molecular mechanisms and biomarkers, and explores their translational potential. Finally, we outline current limitations and future directions toward clinical implementation of pro-teomic findings in precision oncology for uLMS patients.

The emergence of antimicrobial resistance (AMR) in Clostridium difficile (C. difficile), particularly to last-line antibiotics such as linezolid, represents a critical challenge in clinical settings. This study investigates the genomic epidemiology of linezolid-resistant C. difficile, focusing on the distribution and mutational patterns of the chloramphenicol-florfenicol resistance (cfr) gene and its association with multidrug resistance. We analyzed 514 clinical isolates (354 from NCBI Pathogen Detection, 160 from EnteroBase), revealing distinct prevalence patterns among cfr subtypes: cfr(C) was dominant (156/354 NCBI strains; 101/160 EnteroBase strains), whereas cfr(B) frequently harbored missense mutations (p.R247K, p.V294I, and less commonly p.A334T). The cfr(E) subtype was exclusively identified in ribotype 027 (RT027) strains. Notably, cfr(C) exhibited a strong association with RT017, correlating with a conserved 99 bp genomic deletion. Phylogenetic analysis linked cfr-carriage to predominant sequence types (ST1 in NCBI strains, ST37 in EnteroBase isolates). Furthermore, the co-occurrence of cfr with additional AMR genes conferred resistance to macrolides (erythromycin, azithromycin) and tetracyclines, indicating a convergent evolution toward multidrug resistance. These findings underscore the interplay between cfr mutations, hypervirulent ribotypes, and AMR dissemination, necessitating enhanced surveillance to mitigate the spread of resistant C. difficile lineages.

Despite their favorable biopharmaceutical properties, exosomes face standardization challenges in isolation and characterization, hindering the development of EV products suitable for clinical applications. The MRC-5 cell line poses a cellular model approved by regulatory affairs for drug development, facilitating the clinical translation of MRC-5 cell-derived nanotherapeutics. This study aims to: (a) identify the optimal methodology for isolating intact MRC-5 cell-derived exosomes with high purity and integrity suitable for drug delivery applications, (b) elucidate the global miRNA and proteomic profile of the isolated exosomes using high-throughput methodologies and bioinformatics, (c) investigate the exosome uptake capacity towards the "autologous" normal MRC-5, and "heterologous" malignant human lung adenocarcinoma A549 and tongue squamous HSC-3 cells, (d) evaluate the pharmacological profile of doxorubicin (DOX)- and curcumin (CUR)- loaded exosomes towards the normal and tumorigenic cells in vitro. The proposed isolation protocol combining ultrafiltration with membrane-based affinity binding, yielded non-aggregated exosomes with superior physicochemical properties. The miRNA profiling data revealed the enrichment of several tumor-suppressive miRNAs mainly involved in gene regulation pathways, whilst the proteomic profiling highlighted the role of exosomal protein in extracellular matrix remodeling. The drug delivery profiling indicated an "intrinsic" tropism of the exosomes towards the malignant A549 and HSC-3 cells. Exosomal CUR and exosomal DOX inhibited tumor cell proliferation more efficiently than free drugs, with synergistic effects upon co-administration. This study provides a comprehensive morphological, physicochemical and molecular characterization of MRC-5 cell-derived exosomes and validates the "intrinsic" tropism of CUR- and DOX- loaded exosomes towards tumorigenic cells, paving the way towards their further exploitation as drug delivery nanocarriers.

Background/Objectives: Klebsiella pneumoniae ST258 and ST11 are global high-risk antimicrobial-resistant clones known for their virulence and resistance gene dissemination. This study aims to identify these clones in a Greek tertiary hospital and understand their resistance profiles and transmission dynamics. Methods: In January 2025, we isolated two distinct carbapenem-resistant K. pneumoniae in a Greek tertiary hospital: INT18S from an ICU patient’s bronchioalveolar lavage and INT20U from a urine sample in the emergency unit. Antimicrobial susceptibility testing (via Microscan system) and Whole-Genome Sequencing (WGS) were conducted on both isolates and their genomes were submitted to the NCBI. Results: The INT18S isolate carried the blaKPC-2 gene and belonged to the ST258 clone. The INT20U isolate carried the blaNDM-1 gene and belonged to the ST11 clone lineage. Both isolates contained at least one of the extended spectra β-lactamase genes tested (TEM, SHV, OXA-1 and CTX-M group). Conclusions: The co-existence of the high-risk K. pneumoniae clones ST258 and ST11 in different hospital departments increases the risk of resistance gene transfer and suggests potential intra-hospital transmission pathways. Understanding their resistance profiles is critical for guiding treatment strategies and preventing the spread of multidrug-resistant pathogens.

Background: Multi-epitope vaccines have become the preferred strategy for protection against infectious diseases by integrating multiple MHC-restricted T-cell and B-cell epitopes that elicit both humoral and cellular immune responses against pathogens. Computational methods address various aspects independently, yet their orchestration is technically challenging, as most bioinformatics tools are accessible through heterogeneous interfaces and lack interoperability features. The present work proposes a novel framework for rationalized multi-epitope vaccine design that streamlines end-to-end analyses through an integrated web-based environment. Results: VaccineDesigner is a comprehensive web-based framework that streamlines the design of protective epitope-based vaccines by seamlessly integrating computational methods for B-cell, CTL, and HTL epitope prediction. VaccineDesigner incorporates single-epitope prediction and evaluation as well as additional analyses, such as multi-epitope vaccine generation, estimation of population coverage, molecular mimicry, and proteasome cleavage. The functionalities are transparently integrated into a modular architecture, providing a single access point for rationalized, multi-epitope vaccine generation in a time- and cost-effective manner. Conclusions: VaccineDesigner is a web-based tool that identifies and evaluates candidate B-cell, CTL, and HTL epitopes and constructs a library of multi-epitope vaccines that combine strong immunogenic responses, safety, and broad population coverage. The source code is available under the academic license and freely accessible.

Bacillus species are among the most promising plant growth-promoting bacteria (PGPB) due to their adaptability to various environmental niches and extensive biosynthetic capabilities. Despite the available data on the PGP-traits (PGPTs) of Bacillus, the genetic basis underlying their beneficial effects remains largely unexplored. In this study, a comparative genomic analysis of three B. pumilus and one B. pseudomycoides strains, isolated from the maize rhizosphere, is presented to elucidate the molecular mechanisms behind their PGP-traits. All strains exhibited multiple PGP-traits, including phosphate solubilization, phytohormone and siderophore production, growth in nitrogen-free medium, stress tolerance, and biofilm formation. Phylogenomic analysis revealed that plant-associated strains have higher genetic similarity, emphasizing niche-specific evolution. Genome analyses revealed strain- and species-specific adaptations, particularly in relation to nutrient acquisition and abiotic stress response mechanisms. B. pumilus strains encoded alternative sigma factors (SigB, SigM, SigW) enabling enhanced salt tolerance, whereas B. pseudomycoides lacked this system and relied on conventional osmoprotective strategies. The strains utilized different tryptophan-dependent (IAN, IAM or IPyA) pathways for auxin biosynthesis and differed in phosphate solubilization ability, which can be attributed to upstream and missense variants in genes affecting acid metabolism (gltA, acnA, acnB, citM, and citS) and phosphatase (phoA) activity. Iron uptake via bacillibactin-siderophores was exclusive to B. pumilus. The inability of the B. pseudomycoides strain to acquire iron was associated with structural variants (absence of bsaA gene) within the bacillibactin biosynthetic gene cluster. This work provides new insights into the molecular basis of PGP traits in Bacillus and supports the development of Bacillus-based bioinoculants for sustainable agriculture.

Machine learning and genomic medicine are the mainstays of research in delivering personalized healthcare services for disease diagnosis, risk stratification, tailored treatment, and prediction of adverse effects. However, potential prediction errors in healthcare services can have life-threatening impact, raising reasonable skepticism about whether these applications have practical benefit in clinical settings. Conformal prediction offers a versatile framework for addressing these concerns by quantifying the uncertainty of predictive models. In this perspective review, we investigate potential applications of conformalized models in genomic medicine and discuss the challenges towards bridging genomic medicine applications with clinical practice. We also demonstrate the impact of a binary transductive model and a regression-based inductive model in predicting drug response as well as the performance of a multi-class inductive predictor in addressing distribution shifts in molecular subtyping. The main conclusion is that as machine learning and genomic medicine are increasingly infiltrating healthcare services, conformal prediction has the potential to overcome the safety limitations of current methods and could be effectively integrated into uncertainty-informed applications within clinical environments.

Healthcare workplaces lack vital digital skills. In 2023, co-design sessions identified 20 essential learning objectives (LOs) aligned with EU policies.

Background/Objective: Predicting pharmacological response in cancer remains a key challenge in precision oncology due to intertumoral heterogeneity and the complexity of drug–gene interactions. While machine learning models using multi-omics data have shown promise in predicting pharmacological response, selecting the features with the highest predictive power critically affects model performance and biological interpretability. This study aims to compare computational and biologically informed gene selection strategies for predicting drug response in cancer cell lines and to propose a feature selection strategy that optimizes performance. Methods: Using gene expression and drug response data, we trained models on both data-driven and biologically informed gene sets based on the drug target pathways to predict IC50 values for seven anticancer drugs. Several feature selection methods were tested on gene expression profiles of cancer cell lines, including Recursive Feature Elimination (RFE) with Support Vector Regression (SVR) against gene sets derived from drug-specific pathways in KEGG and CTD databases. The predictability was comparatively analyzed using both AUC and IC50 values and further assessed on proteomics data. Results: RFE with SVR outperformed other computational methods, while pathway-based gene sets showed lower performance compared to data-driven methods. The integration of computational and biologically informed gene sets consistently improved prediction accuracy across several anticancer drugs, while the predictive value of the corresponding proteomic features was significantly lower compared with the mRNA profiles. Conclusions: Integrating biological knowledge into feature selection enhances both the accuracy and interpretability of drug response prediction models. Integrative approaches offer a more robust and generalizable framework with potential applications in biomarker discovery, drug repurposing, and personalized treatment strategies.

The therapeutic potential of cold physical gas plasma operated at atmospheric pressure in oncology has been thoroughly demonstrated in numerous preclinical studies. The cytotoxic effect on malignant cells has been attributed mainly to biologically active plasma-generated compounds, namely, reactive oxygen and nitrogen species. The intracellular accumulation of reactive oxygen and nitrogen species interferes strongly with the antioxidant defense system of malignant cells, activating multiple signaling cascades and inevitably leading to oxidative stress-induced cell death. This study aims to determine whether plasma-induced cancer cell death operates through a universal molecular mechanism that is independent of the cancer cell type. Using whole transcriptome data, we sought to investigate the activation mechanism of plasma-treated samples in patient-derived prostate cell cultures, melanoma, breast, lymphoma, and lung cancer cells. The results from the standardized single-cohort gene expression analysis and parallel multi-cohort meta-analysis strongly indicate that plasma treatment globally induces cancer cell death through immune-mediated mechanisms, such as interleukin signaling, Toll-like receptor cascades, and MyD88 activation leading to pro-inflammatory cytokine release and tumor antigen presentation.

The disruption of gut microbiota caused by antibiotics favors the intestinal colonization of Clostridioides difficile - a Gram-positive, spore-forming anaerobic bacterium that causes potentially fatal gastrointestinal infections. In an endeavor to elucidate the complexities of the gut-brain axis in the context of Clostridium difficile infection (CDI), a murine model has been used to investigate the potential effects of antibiotic administration and subsequent colonization by C. difficile, as well as the impact of three different 10-day treatments (metronidazole, probiotics, and fecal microbiota transplantation), on the cecal metabolome for the first time. This follows our previous research which highlighted the metabolic effect of CDI and these treatments in the brain and employs the same four different metabolomics-based methods (targeted GC-MS/MS, targeted HILIC-MS/MS, untargeted RP-LC-HRMS/MS and untargeted GC-MS). A total of 286 unique metabolites have been identified in the mouse cecal profiles and statistical analysis revealed that CDI, as well as the subsequent treatments, significantly alters cecal metabolites and lipids implicated in various biochemical pathways centered around amino acid metabolism, glycerophospholipid metabolism, and central carbon metabolism. To our knowledge, this study represents the first exploration of the effects of C. difficile-induced colitis and potential treatments on the cecal tissue metabolome.

This research evaluated the impact of incorporating dried olive pulp (OP) into the feed of laying hens on the fatty acid profile, cholesterol, triglyceride, total phenolic, oleuropein and hydroxytyrosol content, and health lipid indices of eggs produced by mid- (39 weeks) and late-laying (59 weeks) birds. Over a 36-week trial, 300 eggs from 180 Isa-Brown hens, assigned to three dietary groups with different OP levels (CON, OP4 and OP6), were analyzed. OP reduced egg cholesterol, with significant effects in late-age eggs (p < 0.05). In mid-age hens, the OP6 eggs had higher total phenolics than the controls (p < 0.05) and more PUFAs than the other groups (p < 0.05). The concentration of total phenolics, cholesterol, n3 PUFAs and % fat increased with hen age (p < 0.05), while triglycerides and oleuropein decreased (p < 0.05). With increasing hen age, the SFAs in the OP eggs decreased (p < 0.05) and the MUFAs increased (p < 0.05). Eggs from older hens had higher nutritional value, as indicated by the lower n6/n3 PUFA ratio, lower AI and TI indices, and higher h/H ratio (p < 0.05). Overall, dietary OP supplementation improved the nutritional quality of eggs, suggesting potential health benefits. Our results also highlighted eggs from older hens as a valuable source of high-quality fats.

This study examines the dietary effect of dried olive pulp (OP) on the overall performance, egg quality, health, and gut microbiota of laying hens during a 36-week trial. A total of 180 Isa Brown layers, aged 23 weeks, were assigned to 15-floor pens and divided into three feeding groups (CON, OP4, and OP6) based on the dietary level of OP. Egg quality and biochemical parameters were assessed in 39- and 59-week-old hens. Fecal samples were collected for microbiota analysis. Data were analyzed with an Analysis of Variance. The percentage of broken eggshells was found to be 15–34% lower in the OP groups compared to the CON groups. At 59 weeks of age, a significant reduction in shell thickness was observed in the CON eggs compared to the OP eggs (p < 0.05). At 39 weeks of age, OP6 eggs had the darkest yolk color of all groups (p < 0.05). Fecal microbial diversity was affected only by hens’ age. However, an enrichment in bacterial species belonging to the genera Megasphaera and Megamonas was found in the OP groups at 59 weeks of age. Our results demonstrate that OP feeding beneficially affects egg quality and promotes the proliferation of bacteria involved in the degradation of complex plant compounds, potentially contributing to the overall health of the gut microbiota.

Carbapenemase-producing Pseudomonas aeruginosa strains present a specific geographical distribution regarding the type of carbapenemase-encoding genes that they harbor. For more than twenty years, VIM-type enzymes were the only major carbapenemases that were detected among P. aeruginosa isolates in Greece until the emergence of NDM-1-encoding P. aeruginosa in early 2023. In the present study, we present the rapid reversal of the carbapenemase-producing P. aeruginosa epidemiology from blaVIM- to blaNDM-harbouring isolates that occurred in our hospital since then. Between January 2023 and February 2024, 139 isolates tested positive for carbapenemase production with the NG-Test CARBA 5 immunochromatographic assay. Eight isolates were processed with the Hybrispot antimicrobial resistance direct flow chip molecular assay, and the first NDM-producing isolate was further analyzed through whole genome sequencing and bioinformatics analysis. Multiple resistance genes were detected by molecular techniques in accordance with the extensively drug-resistant phenotype. The isolate that was subjected to whole-genome sequencing belonged to the P. aeruginosa high-risk clone ST308, and the blaNDM was located in the chromosome in accordance with previously reported data. During the study period, NDM-producing isolates were increasingly detected, and only five months after their emergence, they overcame VIM producers. Our results indicate the potential of this new clone to spread rapidly and predominate within healthcare institutions, further restricting the already limited treatment options.

Background/Objectives: Exosomes, nano-sized extracellular vesicles released by all cells, play a key role in intercellular communication and carry tumorigenic properties that impact surrounding or distant cells. The complexity of the exosomal molecular interactome and its effects on recipient cells still remain unclear. This study aims to decipher the molecular profile and interactome of lung adenocarcinoma A549 cell-derived exosomes using multi-omics and bioinformatics approaches. Methods: We performed comprehensive morphological and physicochemical characterization of exosomes isolated from cell culture supernatant of A549 cells in vitro, using DLS, cryo-TEM, Western blot, and flow cytometry. Proteomic and miRNA high-throughput profiling, coupled with bioinformatics network analysis, were applied to elucidate the exosome molecular cargo. A comparative miRNA analysis was also conducted with exosomes derived from normal lung fibroblast MRC-5 cells. Results: Exosomes exhibited an average size of ~40 nm and disk-shaped lipid bilayer structures, with tetraspanins CD9 and CD63 validated as exosomal markers. Proteomic analysis identified 68 proteins, primarily linked to the extracellular matrix organization and metabolic processes. miRNA sequencing revealed 72 miRNAs, notably hsa-miR-619-5p, hsa-miR-122-5p, hsa-miR-9901, hsa-miR-7704, and hsa-miR-151a-3p, which are involved in regulating metabolic processes, gene expression, and tumorigenic pathways. Th integration of proteomic and miRNA data through a proteogenomics approach identified dually affected genes including ERBB2, CD44, and APOE, impacted by both exosomal miRNA targeting and protein interactions through synergistic or antagonistic interactions. Differential analysis revealed a distinct miRNA profile in A549 exosomes, associated with cancer-related biological processes, compared to MRC-5 exosomes; notably, hsa-miR-619-5p emerged as a promising candidate for future clinical biomarker studies. The network analysis also revealed genes targeted by multiple upregulated tumor-associated miRNAs in potential exosome-recipient cells. Conclusions: This integrative study provides insights into the molecular interactome of lung adenocarcinoma A549 cell-derived exosomes, providing a foundation for future research on exosomal cargo and its role in tumor cell communication, growth, and progression.

In the current study, the genotypic characteristics such as antimicrobial resistance and virulence genes, and plasmid replicons and phenotypic characteristics such as biofilm formation and antimicrobial resistance of 87 extended-spectrum beta-lactamase (ESBL)-producing E. coli (ESBL-Ec) isolated from 7 water bodies in northern Greece were investigated. Our data show a high prevalence (60.0 %) of ESBL-Ec in surface waters that exhibit high genetic diversity, suggesting multiple sources of their transmission into the aquatic environment. When evaluating the antimicrobial resistance of isolates, wide variation in their resistance profiles has been detected, with all isolates being multi-drug resistant (MDR). Regarding biofilm formation capacity and phylogenetic groups, the majority (54.0 %, 47/87) of ESBL-Ec were classified as no biofilm producers mainly assigned to phylogroup A (35.6 %; 31/87), followed by B2 (26.5 %; 23/87). PCR screening showed that a high proportion of the isolates tested positive for the blaCTX-M-1 group genes (69 %, 60/87), followed by blaTEM (55.2 %, 48/87), blaOXA (25.3 %, 22/87) and blaCTX-M-9 (17.2 %, 15/87). A subset of 28 ESBL-Ec strains was further investigated by applying whole genome sequencing (WGS), and among them, certain clinically significant sequence types were identified, such as ST131 and ST10. The corresponding in silico analysis predicted all these isolates as human pathogens, while a significant proportion of WGS-ESBL-Ec were assigned to extraintestinal pathogenic E. coli (ExPEC; 32.1 %), and urinary pathogenic E. coli (UPEC; 28.6 %) pathotypes. Comparative phylogenetic analysis, showed that the genomes of the ST131-O25:H4-H30 isolates are genetically linked to the human clinical strains. Here, we report for the first time the detection of a plasmid-mediated mobile colistin resistance gene in ESBL-Ec in Greece isolated from an environmental source. Overall, this study underlines the role of surface waters as a reservoir for antibiotic resistance genes and for presumptive pathogenic ESBL-Ec.