Search Results (7,296)

The efficacy of enzyme therapy is limited by their poor stability under physiological conditions. Thermostable enzymes, derived from extremophilic organisms or generated by advanced protein engineering, offer a revolutionary solution to this long-standing challenge. They are widely used in industrial biocatalysis. Their therapeutic applications are poorly investigated and spread across diverse disciplines. While most applications are in the preclinical stages, emerging evidence from animal models demonstrates proof-of-concept for thermostable antioxidant enzymes in cardiovascular, neurodegenerative, and inflammatory diseases. This review critically assesses the translational landscape, distinguishing between established therapeutic enzymes (e.g., asparaginase, PEGylated SOD) and emerging experimental candidates. This narrative review consolidates existing knowledge about thermostable enzyme engineering and their emerging functions as molecular therapies, particularly in oxidative stress-related diseases. This review synthesizes recent advances in structural biology, computational protein design, biomaterials engineering, and translational antioxidant strategies, highlighting how breaking down disciplinary barriers is accelerating the development of sustainable and self-regenerating antioxidant platforms. By integrating molecular precision with systems-level therapeutic design, engineered thermostable antioxidant enzymes exemplify the future of biological development, where multidisciplinary collaboration drives innovation against oxidative stress-driven pathologies. Engineered thermostable enzymes provide a versatile basis for next-generation therapeutics, with the potential to address medical needs through improved stability, targeted activity, and multifunctional design. Full article

Hepatocellular carcinoma (HCC) is a lethal malignancy requiring novel therapeutic interventions. While Polyporus umbellatus exhibits anti-tumor properties, its specific bioactive pharmacophores and molecular mechanisms remain elusive. This study integrated network pharmacology, computational simulation, and experimental validation to decipher the anti-HCC efficacy of Polyporus umbellatus. Screening identified 11 bioactive sterols, with intersection analysis revealing 63 core targets. Clinical data stratified Checkpoint Kinase 1 (CHEK1) as a critical high-risk oncogene associated with poor prognosis. Molecular dynamics simulations (100 ns) demonstrated that polyporusterone E, a key constituent, forms a thermodynamically stable complex with CHEK1 via high-affinity hydrogen bonding. In vitro assays in HepG2 and HuH-7 cells confirmed that CHEK1 overexpression drives proliferation and metastasis, while its silencing reverses these phenotypes. Crucially, treatment with Polyporus umbellatus extract and purified polyporusterone E significantly compromised HCC cell viability and downregulated CHEK1 expression at transcriptional and translational levels. These findings suggest that polyporusterone E may downregulate CHEK1 expression and modulate CHEK1-associated signaling in HCC cells, providing preliminary evidence for the molecular basis of Polyporus umbellatus and highlighting its potential as a complementary therapeutic strategy for HCC management. Full article

Colon cancer remains a leading cause of cancer-related deaths, and drug repurposing offers a promising strategy to identify new therapies. Hydroquinidine (HQ), a class I antiarrhythmic agent, has recently been suggested to possess anticancer properties; however, its preclinical safety and efficacy in colorectal cancer are not well defined. The safety of HQ was evaluated in Wistar rats following OECD guidelines. Rats received daily intraperitoneal doses (2.5–25 mg/kg) for 90 days, with hematological, biochemical, and histopathological assessments performed. HQ was well tolerated up to 12.5 mg/kg, whereas 25 mg/kg caused signs of hepatotoxicity without lethality. A 1,2-dimethylhydrazine-induced colorectal cancer model was then used to assess HQ at safe doses (6.25 and 12.5 mg/kg) compared with cisplatin. Tissue histopathology and selected molecular markers associated with inflammation, apoptosis, epithelial–mesenchymal transition, and PI3K/AKT/mTOR pathway activity were analyzed. In the DMH-induced colon cancer model, HQ improved colonic tissue architecture and was associated with lower histopathological scores compared with untreated tumor controls. HQ also modulated tumor-associated markers by reducing IL-6 immunoreactivity, increasing caspase-3 expression, enhancing E-cadherin immunoreactivity, and decreasing vimentin expression. Moreover, HQ was associated with reduced immunoreactivity of mTOR pathway-related markers, suggesting attenuation of pathway activation in this experimental context. Overall, HQ showed an acceptable safety profile at the selected doses and exerted favorable histopathological and molecular modulatory effects, supporting further investigation as a potential repurposing candidate. Full article

Evolutionary Strategies (ESs) are optimization metaheuristics largely adopted in Evolutionary Computation (EC). Since their introduction in the early 70s, researchers in the field have attempted to improve the efficacy of these algorithms. The most advanced ESs, such as the Covariance Matrix Adaptation Evolutionary Strategy (CMA-ES) and Exponential Natural Evolution Strategies (xNESs), make use of covariance matrices storing relationships between parameters to be optimized, which enable the algorithms to fasten the search in the solution spaces. However, the computational cost of calculating covariance matrices linearly scales with the number of parameters. Recently, the OpenAI Evolutionary Strategy (OpenAI-ES) emerged as an effective ES in different domains, thanks to the parameter information stored in two momentum vectors. Furthermore, OpenAI-ES gains an advantage from the usage of symmetric sampling and weight decay techniques. In this work, I delve into the application of symmetric sampling and weight decay on CMA-ES, xNES and Separable Natural Evolution Strategies (sNESs), with the aim to improve their performance in domains in which they get stuck in local minima outcomes. Specifically, I propose three novel variants for each ES and verify their efficacy with respect to the PyBullet halfcheetah and hopper robot locomotion problems, and two collective tasks (i.e., swarm aggregation and swarm foraging). The findings reveal that symmetric sampling produces performance enhancements in all the domains, whereas the effect of weight decay varies across the considered problems. Furthermore, symmetric sampling allows ESs to keep parameter size limited, which is paramount in these scenarios. This research identifies techniques enhancing the success of modern ESs, proposes several ES variants, and discusses the relationship between algorithmic performance and task properties. Full article

Background: Insomnia and stress-related sleep disorders are increasingly recognized as systemic conditions linked to the microbiota–gut–brain axis (MGBA). With growing clinical interest in natural products that modulate the gut environment, this systematic review evaluates the efficacy and mechanisms of non-pharmacological interventions, specifically probiotics, prebiotics, dietary indices, and botanicals, in alleviating insomnia, restoring circadian rhythms, and modulating neurochemical markers. Methods: In strict accordance with PRISMA 2020 guidelines, we searched PubMed, ScienceDirect, Scopus, and The Cochrane Library for English language studies published from inception to March 31, 2026. Eligibility was restricted to studies with rigorously controlled designs, specifically randomized controlled trials (RCTs) and controlled in vivo animal studies. Interventions had to target the gut microbiota, with primary outcomes measuring sleep quality (subjective or objective) or sleep-related neurochemical markers. We excluded uncontrolled, single-arm, or observational designs; in vitro studies; non-original research; and studies involving subjects with severe medical or psychiatric comorbidities (e.g., cancer, ADHD, severe psychiatric disorders) to prevent confounding variables, though mild-to-moderate anxiety and depression were permitted. Risk of bias was assessed using the Cochrane RoB 2.0 and SYRCLE tools. Due to significant methodological heterogeneity, a narrative synthesis stratified by intervention and population was conducted. This review was not registered in PROSPERO. Results: A total of 56 studies (33 humans, 23 animals) met the inclusion criteria. Taxonomic nomenclature was updated to reflect 2020 reclassifications (e.g., Lactiplantibacillus plantarum). In human trials, interventions significantly improved subjective sleep metrics (PSQI, ISI). Recent additions demonstrated the efficacy of the Dietary Index for Gut Microbiota (DI-GM) and the improvement in N3 sleep latency by yeast mannan. Furthermore, whole-food patterns (e.g., the MIND diet) and Traditional Chinese Medicine (TCM) decoctions successfully enriched beneficial taxa, such as Bacteroides coprophilus, and increased short-chain fatty acid (SCFA) production. Animal models demonstrated that “psychobiotic” strains (Bifidobacterium breve, Lacticaseibacillus paracasei), prebiotics (GOS/PDX), and TCM formulas effectively restored GABA/5-HT profiles, lowered morning cortisol, and facilitated REM rebound in PCPA-induced models, while also consolidating non-rapid eye movement (NREM) sleep and downregulating clock genes (Per1/Per2). Conclusions: Psychobiotics, prebiotics, and botanicals represent a highly viable non-pharmacological strategy for treating insomnia. However, current evidence is constrained by a heavy reliance on subjective human questionnaires, short follow-up durations limiting insight into long-term stability, and a substantial translational gap between mechanistic rodent models and human clinical outcomes. Full article

Background: Anti-amyloid monoclonal antibodies represent the first disease-modifying therapeutic strategy targeting amyloid-β pathology in early Alzheimer’s disease (AD). Although several agents have demonstrated the ability to reduce cerebral amyloid burden, their clinical efficacy and safety remain subjects of substantial scientific and regulatory debate. This study aimed to synthesize randomized evidence evaluating the benefit–risk profile of anti-amyloid monoclonal antibodies in biomarker-confirmed early AD. Methods: A systematic review and classical pairwise meta-analysis of randomized controlled trials (RCTs) was conducted following the PRISMA 2020 guidelines. PubMed/MEDLINE, Embase, and the Cochrane Central Register of Controlled Trials were searched for phase III placebo-controlled trials evaluating lecanemab, donanemab, aducanumab, and gantenerumab in patients with mild cognitive impairment due to AD or mild AD dementia with biomarker confirmation of amyloid pathology. The primary outcome was change from baseline in the Clinical Dementia Rating–Sum of Boxes (CDR-SB) at the longest available follow-up. Safety outcomes included amyloid-related imaging abnormalities with edema or effusion (ARIA-E), amyloid-related imaging abnormalities with hemorrhage (ARIA-H), serious adverse events, and treatment discontinuation. Random-effects meta-analyses were performed. Results: Six randomized comparisons derived from four phase III trials involving 7695 participants met the eligibility criteria. Anti-amyloid monoclonal antibodies were associated with a statistically significant slowing of clinical progression compared with placebo (pooled mean difference in CDR-SB: −0.42 points; 95% CI −0.59 to −0.25; I² = 78%). The observed effect was primarily driven by trials of lecanemab and donanemab, whereas aducanumab demonstrated discordant results across trials and gantenerumab showed no clinically meaningful benefit. Despite statistical significance, the magnitude of the pooled effect approached the lower boundary of the minimal clinically important difference reported for CDR-SB in early AD. Treatment was associated with a markedly increased risk of ARIA-E (pooled risk ratio 10.1; 95% CI 7.8–13.0), with moderate heterogeneity across studies. Most ARIA-E events were asymptomatic and detected through protocol-mandated MRI monitoring. Conclusions: In biomarker-confirmed early Alzheimer’s disease, anti-amyloid monoclonal antibodies produce a statistically significant but modest slowing of clinical decline accompanied by a substantially increased risk of ARIA. The benefit–risk profile appears heterogeneous across individual antibodies and may reflect pharmacological differences in amyloid targeting and clearance mechanisms. These findings support cautious, individualized use of anti-amyloid therapies and highlight the need for longer-term studies to determine whether short-term slowing of decline translates into clinically meaningful disease modification. Full article

Unloading-induced bone loss is a major medical challenge during long-duration human spaceflight, largely driven by suppressed osteoblast-mediated bone formation, and practical countermeasures are needed. Electromagnetic stimulation has shown benefits for bone repair, and its non-invasiveness supports potential space use; however, its single-modality efficacy remains limited. Here, we investigated a combined electromagnetic field (CEMF) integrating a static magnetic field (SMF, 0.4–0.6 T) and a pulsed electromagnetic field (PEMF, 0.38 ± 0.19 mT) to attenuate unloading-related bone loss and examine field-induced mechanical stimulation. Finite-element simulations mapped magnetic flux density, field gradient, induced current density, and Lorentz force density in bone tissue. CEMF was evaluated in vivo in hindlimb unloading (HLU) mice and in vitro in MC3T3-E1 osteoblasts. CEMF improved bone mineral density, trabecular and cortical microarchitecture, and mechanical properties in HLU mice, with increased osteoblast number and mineral apposition rate. In vitro, CEMF promoted osteogenic differentiation and upregulated COL1A1 and RUNX2. Transcriptome analysis suggested activation of ECM–integrin mechanical signaling and the PI3K–AKT pathway. These findings indicate that CEMF-induced multiphysics stimulation enhances osteogenic responses and may serve as a complementary, non-invasive countermeasure for spaceflight-associated bone loss. Full article

Timely detection and monitoring of diseases in sweet potato crops are important for hunger alleviation and food security. This study aimed to evaluate the efficacy of the optimized field spectrometric reflectance thresholds in spatially partitioning sweet potato crops on the unmanned aerial vehicle (UAV) multispectral imagery based on infection types. A field survey was carried out to sample deformed leaves for laboratory diagnosis of possible identification of sweet potato leaf infection types. Laboratory analysis results revealed nutrient deficiency, SPVC-positive, fungi isolates (i.e., alternaria, bipolaris, fusarium, phoma), and mechanical damage as the causes of leaf deformation. Overlap analysis results revealed reflectance overlaps across all leaf deformation types, making it difficult to spatially partition sweet potato crops based on deformation types. Instead, sweet potato crops were spatially partitioned by considering the minimum and maximum thresholds acquired from the whole dataset. Area covered by deformed sweet potato leaves in blue, green, red, red edge and NIR were found to be 11.91%, 28.71%, 43.66%, 46.41% and 30.6% respectively. Coefficient of determination results revealed poor classification results, with R² value of 0.23, 0.19, 0.28, 0.17 and 0.63 for blue, green, red, red edge and NIR respectively. However, the NIR spectral band yielded R² value closer to the acceptable value of 0.7. Full article

Food allergy is an increasingly prevalent global health condition characterized by immune-mediated reactions to dietary antigens and a substantial clinical burden. Growing understanding of IgE-mediated mechanisms has highlighted the central role of type 2 inflammation, effector-cell activation, and impaired immune regulation. These advances have prompted the development of disease-modifying therapies beyond allergen avoidance. This narrative review summarizes recent advances in the therapeutic management of IgE-mediated food allergy. A structured PubMed search was performed to identify clinical trials, randomized studies, and meta-analyses published within the last five years. Both allergen-specific and non-allergen-specific interventions were evaluated. Current evidence supports oral immunotherapy as the most effective strategy for increasing reaction thresholds and inducing desensitization in peanut, milk, and egg allergies. However, safety concerns remain, and sustained unresponsiveness after treatment discontinuation is achieved inconsistently. Sublingual and epicutaneous immunotherapy show improved safety but lower efficacy. Modified allergen approaches, including baked milk and processed peanut products, may improve tolerability and facilitate immune modulation in selected patients. Biologic therapies, particularly anti-IgE agents, demonstrate efficacy both alone and when combined with immunotherapy. Emerging approaches include peptide vaccines, DNA immunization, microbiome-targeted interventions, and early dietary modulation. These strategies may improve durable immune tolerance through personalized, mechanism-based therapeutic approaches. Future progress will depend on optimizing safety, identifying predictive biomarkers, and integrating multimodal approaches to achieve durable immune tolerance. Full article

Ionizing radiation induces reactive oxygen species (ROS) and inflammatory signaling that contribute to both therapeutic efficacy and normal tissue toxicity. While the effects of high-dose radiation are well characterized, responses to low-dose-rate radiation (LDRR) remain inconsistent and are not adequately explained by conventional linear dose–response models. To address this gap, we conducted a narrative review of recent experimental studies across multiple biological systems, including body fluids, joint microenvironments, and reproductive tissues, focusing on redox and immune-related responses under LDRR conditions (dose rates: 0.39–3.49 mGy/h). Literature was identified through PubMed/MEDLINE, Web of Science, and Google Scholar, with emphasis on studies published between 2015 and 2026. These studies demonstrate that LDRR elicits nonlinear, dose-dependent effects that vary across biological compartments and involve coordinated changes in oxidative stress, immune signaling, and metabolic regulation. Based on this synthesis, we propose a unifying framework of nonlinear redox–immune coupling, in which oxidative stress functions as a threshold-dependent regulator and immune responses follow a biphasic trajectory characterized by activation at lower dose rates and attenuation or adaptation at higher levels. These responses are strongly influenced by the local microenvironment, resulting in compartment-specific variability. This integrated perspective supports a shift from dose-centric to systems-level interpretations of radiation biology and provides a basis for improving biomarker development, risk assessment, and therapeutic strategies in chronic low-dose radiation exposure settings. Future research priorities include time-resolved mechanistic studies to define compartment-specific redox thresholds, validation of candidate biomarkers under identical multi-compartment experimental conditions (e.g., GSH/GSSG ratio, 8-OHdG, circulating cytokine panels including IL-10/TNF-α ratio), and integration of subject-specific biological variables (e.g., age, sex, and baseline redox capacity) into predictive models of LDRR response. Full article

Incomplete spatio-temporal (ST) data from sensor networks in precision agriculture often limits environmental modeling and decision-making accuracy. To address this, we propose the Spatio-Temporal Conditional Diffusion Framework (ST-CDF), a generative approach for high-fidelity data reconstruction. The framework’s core is a deep denoising network that integrates a Graph Attention Network (GAT) to explicitly model non-Euclidean spatial correlations, a Differential Attention Transformer to capture abrupt temporal dynamics, and an Inverse Discrete Wavelet Transform (IDWT) module to preserve multi-scale signal details. The generative process is constrained by a physics-informed training objective, which injects known physical laws (i.e., the Penman–Monteith equation for reference evapotranspiration, $E{T}_0$) as an inductive bias, ensuring the imputed data maintains physical consistency. For privacy-preserving deployment on resource-constrained IoT devices, we extend the framework with a Federated Cluster-Guided Distillation (Fed-CGD) strategy. We conducted extensive experiments against established methods on two real-world agricultural datasets. ST-CDF demonstrated improved imputation accuracy across evaluated metrics. Its efficacy was most pronounced in the physically-demanding $E{T}_0$ calculation task, where data imputed by ST-CDF at an 80% missing rate achieved a Root Mean Square Error (RMSE) of 0.3485 and a Coefficient of Determination ($R^2$) of 0.7558, outperforming the baseline models. Furthermore, we explore ST-CDF as an explainable (XAI) framework for active agricultural decision support, demonstrating its utility in performing counterfactual simulations of “what-if” interventions, such as irrigation. The findings highlight ST-CDF as an effective, physically-grounded, and interpretable tool for data-driven scientific computation and precision agriculture. Full article

Salmonella remains one of the most critical zoonotic pathogens in the poultry sector, linked to animal disease, foodborne illness, and the global crisis of antimicrobial resistance (AMR). Poultry acts as a major reservoir, enabling Salmonella transmission from hatchery to retail products through horizontal, vertical, and environmental routes. Despite the use of biosecurity, vaccination, antibiotics, and chemical decontamination, effective and sustainable control across the poultry value chain remains difficult, particularly in the face of rising multidrug-resistant strains and growing consumer concerns over chemical residues. Bacteriophages (phages), viruses that selectively infect and lyse bacteria, have emerged as a promising biological alternative for Salmonella control. Although many studies have reported the effectiveness of phages against bacterial species, including Salmonella, in the poultry industry, reports on their full potential to combat antimicrobial-resistant Salmonella across the entire poultry value chain remain limited. Therefore, this review synthesizes current evidence on the application of phages throughout the poultry value chain, including on-farm interventions, processing plant decontamination, and food packaging and storage. Findings from the reviewed articles indicate over a 90% reduction in Salmonella spp. in poultry farms and post-harvest meat, along with lower mortality in phage-treated groups compared to untreated groups; however, these outcomes depend on several factors (e.g., phage strains, concentrations, application methods, and environmental conditions). Laboratory, pilot, and field studies consistently demonstrate that phage preparations, especially when formulated as cocktails or combined with complementary interventions, can achieve substantial reductions in Salmonella, including antibiotic-resistant serovars, in live birds, eggs, poultry environments, and meat products. Unlike antibiotics and chemical sanitizers, phages act with high specificity, preserving beneficial microbiota and maintaining the sensory and nutritional quality of poultry products. Their safety has been supported by toxicological and genomic assessments, and several phage-based products have obtained regulatory approval, including Generally Recognized as Safe (GRAS) status for food applications in the United States. By integrating efficacy, safety, regulatory, and practical deployment data, this review highlights bacteriophages as a scientifically validated and One Health–aligned tool capable of reducing Salmonella transmission from farm to fork across the poultry value chain, thereby laying the foundation for their future adoption in the poultry industry. Phage-based interventions offer a sustainable pathway to enhance food safety, limit antimicrobial resistance (AMR) dissemination, and strengthen consumer confidence in poultry products. However, the major limitation is the emergence of phage-resistant bacterial strains, as well as the potential involvement of some phages in the transfer of resistance and virulence genes, which could raise public concern. Nevertheless, the use of phage cocktails and whole-genome sequencing, involving tools such as ResFinder and virulence finder, can facilitate the selection of safe phages for application. Full article

In Chile, Eucalyptus globulus stands out as a significant forest species, yielding around 2 million tonnes of bark; this by-product is a valuable source of phenolic compounds. This research evaluated the valorization of E. globulus bark using alkali-assisted extraction (AAE) and obtained extracts intended to protect the wood against fungal degradation and ultraviolet (UV) radiation. The chemical and thermal properties of the extracts were characterized using total phenolic content (TPC), antioxidant capacity, FTIR spectroscopy, LC-LTQ-Orbitrap-MS, and thermal analyses (TGA and DSC). Pine wood samples were impregnated using the Bethel process, and their absorption, retention, leaching, UV resistance, gloss, and antifungal efficacy were evaluated. The AAE showed an extraction yield of 8.79%, almost double that of aqueous extraction, with a phenolic content of 970 mg GAE/100 g dry bark and good antioxidant capacity. The MS/MS analysis tentatively identified low-molecular-weight organic acids, phenolic acids, a hydrolyzable tannin derivative, ellagic acid, methylated flavonol glycosides, and an iridoid non-phenolic metabolite. Thermal analysis indicated greater stability of the alkaline extracts, with a mass loss of less than 10% up to 200 °C, and significant degradation between 220 and 300 °C. Leaching tests showed a lower release of polyphenols from alkali-treated wood, indicating reduced mobility and/or greater retention of the extractives within the wood structure. Biological assays demonstrated effective inhibition of stain fungi and strong resistance to brown rot. Furthermore, UV aging tests showed less color change (Delta E*) and greater resistance to surface degradation. These results demonstrate the potential of alkaline extracts from E. globulus bark as sustainable additives for wood protection. Full article

Background: The high incidence and severe health threat of Toxoplasma gondii (T. gondii) infection, particularly in immunocompromised patients, underscore the urgent need for the development of a safe and effective vaccine. The aim of this study was to develop a novel multi-epitope vaccine (USM.TOXOII) incorporating the T. gondii GRA14, SAG1, and GRA1 antigens, and to assess its immunogenicity in BALB/c mice. Methods: Using bioinformatics approach, the USM.TOXOII was designed and evaluated. The encoding gene (471 bp) was then constructed and cloned into the pET-30a (+) plasmid before being transformed into E. coli expression system. The recombinant USM.TOXOII protein was subsequently expressed and purified. Finally, an animal study was performed to assess the vaccine’s immunogenicity. Results: The USM.TOXOII protein (17.27 kDa) was soluble and contained a His tag protein. Immunization of BALB/c mice with USM.TOXOII significantly elevated serum levels of total IgG, IgG1, and IgG2a (p < 0.05). Cytokine analysis revealed a significant increase in IFN-γ production, whereas IL-4 levels remained unchanged, suggesting a Th1-biased immune response. Conclusions: Collectively, these findings indicate that USM.TOXOII possesses immunogenic potential and is capable of inducing both humoral and cellular immune responses in BALB/c mice. Future challenge studies with live T. gondii tachyzoites are warranted to evaluate its protective efficacy in vivo. Full article

Tropical coastal waters are increasingly recognized as critical reservoirs for virulent, antibiotic-resistant enteric pathogens, yet seasonal dynamics governing their spatial distribution remain poorly characterized. We hypothesized that hydrological shifts and anthropogenic nutrient enrichment drive the seasonal distribution, virulence profiles, and antimicrobial resistance (AMR) of Escherichia coli, Salmonella spp., and Shigella spp. in the Jaffna Peninsula, Sri Lanka. Across 25 coastal sites during dry and transitional seasons, we integrated physicochemical water quality assessment, culture-based enumeration, PCR-based virulence gene profiling, Minimum Inhibitory Concentration (MIC) assays, GIS mapping, and statistical analyses. Key water quality parameters, including ammonium, nitrite, and total phosphorus, showed significant seasonal variation (p < 0.05), reflecting distinct hydrological regimes across seasons. A total of 220 E. coli, 200 Salmonella spp., and 100 Shigella spp. isolates were examined for virulence gene profiles and antibiotic tolerance. E. coli was detected at 80–88% of sites, Salmonella spp. at 72–88%, and Shigella spp. at 32–48%. Among E. coli isolates, stx1 was detected at 20–28% of sites and eae at 16% across both seasons. The stn gene was detected in Salmonella spp. at 12–28% of sites seasonally. Virulence profiling confirmed STEC harbouring stx1, stx2, and eae; Salmonella spp. carried stn; and Shigella spp. possessed invasion-associated genes. Trimethoprim–sulfamethoxazole resistance was recorded in 63.2% of E. coli, 33.0% of Salmonella spp., and 31.0% of Shigella spp. isolates at the lowest tested concentration of 4 µg/mL., while ciprofloxacin and piperacillin–tazobactam retained greater efficacy. Correlation analyses revealed significant associations among faecal contamination, nutrient enrichment, and virulence gene prevalence, implicating untreated sewage discharge and eutrophication as likely ecological factors associated with pathogen occurrence. These findings designate the Jaffna coastal zone as a significant reservoir of virulent AMR enteric pathogens, underscoring the urgent need for integrated One Health surveillance and seasonally adaptive coastal water quality management. Full article