Search Results (514)

(1) Background: Pigs can be another host of Shiga toxin-producing E. coli (STEC), suggesting that pork products could be a potential risk to public health. A USDA National Animal Health Monitoring System (NAHMS) study revealed that Shiga toxin genes were detected in more than half of samples nationwide but only about a quarter of samples from the state of Illinois. To characterize the presence of STEC in Illinois pigs better and to explore the discrepancy between Illinois and other swine-producing states, we increased the sampling size and collected samples in different regions of the state and in different months to detect Shiga toxin genes in Illinois finisher pigs and subtyped the Shiga toxin genes further to assess any potential risk of STEC originating from Illinois pigs to human health. (2) Methods: Fecal samples were collected from 471 Illinois finisher pigs at different locations from October 2021 to September 2022. DNA samples were extracted from individual fecal samples and PCR-tested for Shiga toxin genes (stx1, stx2) and then toxin subtypes (stx2a, stx2c, stx2d, and stx2e). (3) Results: The data showed that the stx2 gene was detected in 61% of the fecal samples (285/471), whereas stx1 was detected only in 0.4% of the samples (2/471). The data also indicated a lower prevalence of stx genes in the samples collected in certain cold months (36% in October and 19% in March) compared to that in those from warm months (56% to 100% from April to September). Stx2d, a subtype associated with severe human illness, was detected in 2% of the samples (10/471); in contrast, stx2e, which is expressed by E. coli strains causing diarrhea and edema disease in pigs, was the most detected (49%; 229/471). (4) Conclusions: The high prevalence of Shiga toxin genes in the fecal samples from Illinois finisher pigs suggests that Stx-positive E. coli strains circulate in Illinois pig farms. However, the highly detected stx2e-positive STEC (or enterotoxigenic E. coli, ETEC) strains are associated with diarrhea and edema disease in pigs, indicating the need for disease prevention or control for pigs but unlikely a safety concern for Illinois pork products or a major risk of human illnesses. Full article

Whole-genome sequence (WGS) analysis was used in this study to characterize Shiga toxin-producing Escherichia coli (STEC) isolates in free-ranging red deer from the central Italian Alps. Fecal samples from 92 hunted red deer collected between September and December 2022 were analyzed for the presence of STEC. Single E. coli colonies positive by PCR for stx genes were analyzed by WGS. STEC were isolated from eleven (12%) samples, showing eight stx2b, one stx2a, two stx1c, and one stx1a subtypes. Different serotypes and sequence types were identified (n = 8 each). Three isolates of O27:H30 serotype and ST753 showed no correlation in the cgMLST analysis (AD range 44–98). All strains harbored additional virulence factors. The only isolate harboring stx2a also possessed the eae gene and belonged to serotype O26:H11. Some isolates displayed shuffled virulence features of more than one E. coli pathotype. The high genetic diversity of strains circulating in the red deer population living in the central Italian Alps, including the STEC O26:H11 strain associated with STEC from severe disease in humans, confirms red deer as STEC reservoirs and highlights the need for monitoring the presence of these pathogens in wild ruminants. Full article

Shiga toxins (Stx), produced by Shiga toxin-producing Escherichia coli, preferentially attack renal tissue and frequently induce acute kidney injury (AKI) and renal failure. To prevent irreversible damage, the injured renal tissue, particularly renal tubular epithelium, mounts a remodeling and regeneration response to repair itself. However, how such intrinsic renal repair processes are initiated and coordinated in infected renal tubular regions remains elusive. Herein, we reported that macula densa apparatus, in addition to its conventional role as a salt sensor in nephron, can function as an endogenous sensor for exogenous toxins (e.g., Stx). We demonstrated that macula densa cells orchestrate a rapid repair niche by initiating transcriptional activation of repair and regeneration factors in both Stx-injured murine models and human kidney organoids. Mechanistically, we showed that in response to Stx exposure, macula densa cells release a specific repair factor CCN1, which effectively promotes the regeneration of toxin-injured renal tubular epithelium and facilitates renal tubular repair through integrin-mediated signaling pathways. Moreover, we demonstrated that treatment with recombinant CCN1 can greatly ameliorate the structural damage and significantly restore the proximal tubular reabsorption capacity in Stx-infected kidney organoids. Our finding highlights a novel role of macula densa apparatus in toxin-induced renal injury, and paves a new avenue for treatment of AKI-associated renal diseases. Full article

Background/Objectives: Escherichia coli (E. coli) strains harboring virulence genes and antimicrobial resistance (AMR) pose a significant risk to poultry production and public health in Pakistan. This study aimed to isolate E. coli from poultry meat and poultry farm environments and compare their virulence gene profiles and AMR patterns. Methods: A total of 100 samples were collected, including 50 poultry meat samples from retail shops and 50 environmental samples from poultry farms. E. coli was isolated on MacConkey agar following overnight enrichment in lactose broth. Isolates were confirmed by biochemical testing and 16S rRNA gene PCR. Virulence genes (stx1, stx2, eae) were detected using multiplex PCR, and AMR profiles were assessed via the Kirby–Bauer disk diffusion method. Results: E. coli was isolated from 26 poultry meat samples (52%) and 23 poultry farm environment samples (46%). All isolates harbored at least one virulence gene, with stx2 being the most prevalent (34.62% meat; 39.13% environment), followed by stx1 (19.23% meat; 17.40% environment) and eae (11.54% meat; 13.04% environment). Combined gene patterns (stx1/eae, stx2/eae, stx1/stx2/eae) were also detected across both sources. AMR analysis revealed high resistance to cefoxitin (100% both sources), trimethoprim (57.09% meat; 60.87% environment), and ampicillin–sulbactam (42.3% meat; 52.17% environment). In contrast, isolates were completely susceptible to norfloxacin (100% meat; 95.65% environment) and exhibited high susceptibility to tetracycline (84.62% meat; 82.61% environment). Statistical comparisons using Fisher’s exact test and the Kruskal–Wallis test showed no significant differences (p > 0.05) in virulence gene prevalence or AMR patterns between poultry meat and environmental isolates. Conclusions: These findings highlight poultry farm environments as potential reservoirs for pathogenic, antimicrobial-resistant E. coli, emphasizing the risk of zoonotic transmission through contaminated poultry meat and the need for improved biosecurity measures. Full article

Saxitoxin (STX) is a toxin with paralyzing and lethal properties, necessitating the development of a simple analytical method. This study developed a nucleic acid aptamer biosensor using graphene oxide (GO) as a fluorescence quencher for STX detection. GO was combined with M30-f, an STX nucleic acid aptamer modification with 5-carboxyfluorescein, which can produce fluorescence absorption under the conditions of an excitation wavelength of 408 nm and emission wavelength of 515 nm. Based on the principle of fluorescence resonance energy transfer, the fluorescence of M30-f was quenched. In the presence of STX, M30-f specifically binds to STX and dissociates from the GO surface, thereby restoring fluorescence. The STX content can be quantitatively detected through differences in fluorescence absorption. The influence of ultrasonic time on the fluorescence quenching ability of GO was investigated. The aqueous solution of graphene oxide, 30GO, optimized by ultrasound treatment for a duration of 30 min, demonstrated excellent fluorescence quenching capability. 30GO was analyzed utilizing various characterization techniques, including SEM, FT-IR, UV, XPS, XRD, AFM, and contact angle measurements. The methodological validation showed that the established STX sensor exhibits excellent linearity within a concentration range of 10–100,000 ng/L, with a limit of detection (LOD) as low as 0.098 μg/L. In addition, the results further demonstrated the sensor’s high specificity for detecting neurotoxic shellfish toxin STX. The recovery rate for clam samples ranged from 89.12% to 104.71%, while that for oyster samples ranged from 91.20% to 109.65%, with relative standard deviations (RSDs) all below 3%. This aptamer sensor is characterized by its simplicity, high sensitivity, and broad detection range, providing significant technical support for advancing marine biotoxin research. Full article

Osteoarthritis (OA) is an inflammatory joint disease and the leading cause of musculoskeletal disability affecting human and veterinary patients. New therapeutics halting inflammation while preserving joint homeostasis remain a critical need. Voltage-gated sodium (NaV) channels regulate the pro-inflammatory response of macrophages in the synovium, the central driver of joint homeostasis. Neosaxitoxin (NeoSTX) is a phycotoxin that blocks NaV channels, conferring a unique potential to regulate joint inflammation. This study evaluated the safety of intra-articular administration of NeoSTX in horses. Sixteen horses were allocated into two groups (n = 8/each). One group received one intraarticular dose (20 µg/2 mL of saline) of NeoSTX into one tarsocrural joint, while the control group received 2 mL of saline (0.9% NaCl). No differences were observed between groups for systemic or local signs of inflammation, including objective measures of surface temperature and joint effusion. Concentrations of synovial fluid total nucleated and differential cell counts, total protein, glucose, calcium, and 23 cytokines/chemokines measured throughout this study did not differ between treatment groups. In this short-term study, intra-articular NeoSTX injection was shown to be well tolerated and likely safe. Ongoing studies should elucidate the role of NeoSTX in modulating synovial mechanisms of inflammation and its endogenous resolution. Full article

Neurodegeneration is increasingly recognized not as a linear trajectory of protein accumulation, but as a multidimensional collapse of biological organization—spanning intracellular signaling, transcriptional identity, proteostatic integrity, organelle communication, and network-level computation. This review intends to synthesize emerging frameworks that reposition neurodegenerative diseases (ND) as progressive breakdowns of interpretive cellular logic, rather than mere terminal consequences of protein aggregation or synaptic attrition. The discussion aims to provide a detailed mapping of how critical signaling pathways—including PI3K–AKT–mTOR, MAPK, Wnt/β-catenin, and integrated stress response cascades—undergo spatial and temporal disintegration. Special attention is directed toward the roles of RNA-binding proteins (e.g., TDP-43, FUS, ELAVL2), m6A epitranscriptomic modifiers (METTL3, YTHDF1, IGF2BP1), and non-canonical post-translational modifications (SUMOylation, crotonylation) in disrupting translation fidelity, proteostasis, and subcellular targeting. At the organelle level, the review seeks to highlight how the failure of ribosome-associated quality control (RQC), autophagosome–lysosome fusion machinery (STX17, SNAP29), and mitochondrial import/export systems (TIM/TOM complexes) generates cumulative stress and impairs neuronal triage. These dysfunctions are compounded by mitochondrial protease overload (LONP1, CLPP), UPR maladaptation, and phase-transitioned stress granules that sequester nucleocytoplasmic transport proteins and ribosomal subunits, especially in ALS and FTD contexts. Synaptic disassembly is treated not only as a downstream event, but as an early tipping point, driven by impaired PSD scaffolding, aberrant endosomal recycling (Rab5, Rab11), complement-mediated pruning (C1q/C3–CR3 axis), and excitatory–inhibitory imbalance linked to parvalbumin interneuron decay. Using insights from single-cell and spatial transcriptomics, the review illustrates how regional vulnerability to proteostatic and metabolic stress converges with signaling noise to produce entropic attractor collapse within core networks such as the DMN, SN, and FPCN. By framing neurodegeneration as an active loss of cellular and network “meaning-making”—a collapse of coordinated signal interpretation, triage prioritization, and adaptive response—the review aims to support a more integrative conceptual model. In this context, therapeutic direction may shift from damage containment toward restoring high-dimensional neuronal agency, via strategies that include the following elements: reprogrammable proteome-targeting agents (e.g., PROTACs), engineered autophagy adaptors, CRISPR-based BDNF enhancers, mitochondrial gatekeeping stabilizers, and glial-exosome neuroengineering. This synthesis intends to offer a translational scaffold for viewing neurodegeneration as not only a disorder of accumulation but as a systems-level failure of cellular reasoning—a perspective that may inform future efforts in resilience-based intervention and precision neurorestoration. Full article

The impact of Oedema Disease produced by Shiga toxigenic Escherichia coli (STEC) in swine is increasing in some production countries due to increasing limitations on treatment with antimicrobials and zinc oxide, either because of the increased prevalence of multi-resistant strains or because of legal restrictions. The main pathological effect of Shiga toxin 2e is represented by damage to the endothelial cells of the blood vessel walls, leading to liquid extravasation and oedema formation in multiple tissues. These oedemas are generally easily identifiable in acute clinical cases. However, disease caused by Shiga toxin can occur without any externally visible oedema in the pigs, as observed in the subclinical presentation of Oedema Disease. It also causes productive losses, so it is important to identify and/or diagnose cases to set up control measures in order to optimize production and health. This article includes a comprehensive review of lesions and signs caused by Shiga toxin toxicosis in pigs, as well as other insights about the aetiology and epidemiology of STEC in pigs, and the effect of Shiga toxin recombinant toxoid vaccines in reducing these clinical and subclinical signs under field conditions. Full article

Background/Objectives: Increasing levels of antimicrobial resistance (AMR) have recently been observed at the human–domestic animal–wildlife interface. Wild birds have been identified as carriers of antimicrobial-resistant bacteria and serve as excellent biomarkers for epidemiological studies. This study assessed the current AMR presence in Eastern Spain’s commensal Escherichia coli isolated from free-ranging Bonelli’s eagles (Aquila fasciata). Methods: Nestlings and their nests were intensively sampled between 2022 and 2024 to determine their AMR profile and characterize E. coli. AMR testing was conducted using the broth microdilution method, following the European Committee on Antimicrobial Susceptibility Testing guidelines. Additionally, the presence of eaeA (intimin gene) and stx-1 and stx-2 (shiga toxins) was analyzed by real-time PCR to classify E. coli strains into enteropathogenic (EPEC) and Shiga-toxigenic (STEC) pathotypes. Results: Of all E. coli isolates, 41.7% were resistant to at least one antimicrobial, and 30% were multidrug-resistant. Only two strains were classified as EPEC and none as STEC. The highest resistance rates were observed for amoxicillin and tetracycline (19.6% each). Alarmingly, resistance to colistin and meropenem, last-resort antibiotics in human medicine, was also detected. Conclusions: Although the mechanisms of resistance acquisition remain unclear, transmission is likely to occur through the food chain, with synanthropic prey acting as intermediary vectors. These results highlight the role of Bonelli’s eagles as essential sentinels of environmental AMR dissemination, even in remote ecosystems. Strengthening One Health-based surveillance is necessary to address AMR’s ecological and public health risks in wildlife. Full article

This study presents a comprehensive comparison of static and SLAM (Simultaneous Localization and Mapping) laser scanners of both new and old generation in a controlled indoor environment of a standard commercial building with long, linear corridors and recesses. The aim was to assess both global and local accuracy, as well as noise characteristics, of each scanner. Methods: A highly accurate static scanner was used to generate a reference point cloud. Five devices were evaluated: two static scanners (Leica RTC 360 and Trimble X7) and three SLAM scanners (GeoSLAM ZEB Horizon RT, Emesent Hovermap ST-X, and FARO Orbis). Accuracy analysis included systematic and random error assessment, axis-specific displacement evaluation, and profile-based local accuracy measurements. Additionally, noise was quantified before and after data smoothing. Static scanners yielded superior accuracies, with the Leica RTC 360 achieving the best performance (absolute accuracy of 1.2 mm). Among SLAM systems, the Emesent Hovermap ST-X and FARO Orbis—both newer-generation devices—demonstrated significant improvements over the older-generation GeoSLAM ZEB Horizon RT. After smoothing, the noise levels of these new-generation SLAM scanners (approx. 2.1–2.2 mm) approached those of static systems. The findings underline the ongoing technological progress in SLAM systems, with the new-generation SLAM scanners becoming increasingly viable alternatives to static scanners, especially when speed, ease of use, and reduced occlusions are prioritized. This makes them well-suited for rapid indoor mapping applications, provided that the slightly lower accuracy is acceptable for the intended use. Full article

Background: Diabetic foot ulcers (DFUs) are a severe complication of diabetes and are characterized by impaired wound healing and a high amputation risk. Exosomes—which are nanovesicles carrying proteins, RNAs, and lipids—mediate intercellular communication in wound microenvironments, yet their biomarker potential in DFUs remains underexplored. Methods: We analyzed transcriptomic data from GSE134431 (13 DFU vs. 8 controls) as a training set and validated findings in GSE80178 (6 DFU vs. 3 controls). A sum of 7901 differentially expressed genes (DEGs) of DFUs were detected and intersected with 125 literature-curated exosome-related genes (ERGs) to yield 51 candidates. This was followed by GO/KEGG analyses and a PPI network construction. Support vector machine–recursive feature elimination (SVM-RFE) and the Boruta random forest algorithm distilled five biomarkers (DIS3L, EXOSC7, SDC1, STX11, SYT17). Expression trends were confirmed in both datasets. Analyses included nomogram construction, functional and correlation analyses, immune infiltration, GSEA, gene co-expression and regulatory network construction, drug prediction, molecular docking, and RT-qPCR validation in clinical samples. Results: A nomogram combining these markers achieved an acceptable calibration (Hosmer–Lemeshow p = 0.0718, MAE = 0.044). Immune cell infiltration (CIBERSORT) revealed associations between biomarker levels and NK cell and neutrophil subsets. Gene set enrichment analysis (GSEA) implicated IL-17 signaling, proteasome function, and microbial infection pathways. A GeneMANIA network highlighted RNA processing and vesicle trafficking. Transcription factor and miRNA predictions uncovered regulatory circuits, and DGIdb-driven drug repurposing followed by molecular docking identified Indatuximab ravtansine and heparin as high-affinity SDC1 binders. Finally, RT-qPCR validation in clinical DFU tissues (n = 5) recapitulated the bioinformatic expression patterns. Conclusions: We present five exosome-associated genes as novel DFU biomarkers with diagnostic potential and mechanistic links to immune modulation and vesicular transport. These findings lay the groundwork for exosome-based diagnostics and therapeutic targeting in DFU management. Full article

A single domain antibody (SDAb) targeting canine PD-1 was developed as a potential immunotherapeutic for canine cancer. An alpaca was immunized with canine PD-1 protein, and a phage-display library was constructed using mRNA isolated from peripheral lymphocytes. Screening of the library yielded multiple SDAb candidates capable of nanomolar binding to canine PD-1. Among these, clone STX-1b5 demonstrated high expression in a yeast-based recombinant system and was selected for further characterization. Binding and competition assays using ELISA confirmed its ability to bind canine PD-1 and block PDL-1 interaction. In silico structural modeling supported the interaction of STX-1b5 with key PD-1 residues implicated in ligand binding. These findings support the feasibility of using SDAbs and cost-effective yeast expression systems to generate immunotherapeutics for veterinary use, with STX-1b5 representing a promising lead candidate for future clinical development. Full article

Subclinical mastitis (SCM) is a stealthy but devastating challenge in the dairy industry, leading to economic losses and hindering efforts to achieve milk self-sufficiency. This study investigated the prevalence of SCM, antimicrobial resistance, and virulence profiles of Escherichia coli. A total of 174 milk samples were analyzed using the California mastitis test (CMT), somatic cell counts (SCCs), bacteriological culture, MALDI-TOF MS, and polymerase chain reaction (PCR). The findings revealed that the SCM prevalence was 68/174 (39.08%) based on CMT and SCC. Among SCM-positive samples, 60/68 (88.23%) were identified as E. coli, confirmed by MALDI-TOF MS and PCR assay. The most frequently detected serogroups were 0113 (11.6%) and 0113 (3.3%). Additionally, the genes for Stx1 and Stx2 were also detected in nine (15%) and one (1.7%), respectively. Antimicrobial susceptibility tests showed widespread resistance, with E. coli isolates demonstrating resistance to penicillin in 43 (71.6%), followed by ciprofloxacin in 42 (70%) and gentamicin in 18 (30%). A larger proportion of the E. coli strains (100%) harbored the bla_VIM gene, while 23 (38.3%), 20%, 20%, and 1.47% contained bla_KPC, bla_NMD, suli1, and msrA. Thirty (50%) isolates were considered multidrug-resistant (MDR). These findings underscore the urgent need for enhanced surveillance and antibiotic stewardship in dairy farming. The presence of MDR E. coli in SCM poses a dual threat of potential transmission to humans and treatment failures in mastitis management. This study highlights the importance of proactive control strategies to mitigate the spread of antimicrobial resistance in livestock and beyond. Full article

Saxitoxin (STX) is a potent toxin produced by marine dinoflagellates and freshwater or brackish water cyanobacteria, and is a member of the paralytic shellfish toxins (PSTs). As a highly specific blocker of voltage-gated sodium channels (NaVs), STX blocks sodium ion influx, thereby inhibiting nerve impulse transmission and leading to systemic physiological dysfunctions in the nervous, respiratory, cardiovascular, and digestive systems. Severe exposure can lead to paralysis, respiratory failure, and mortality. STX primarily enters the human body through the consumption of contaminated shellfish, posing a significant public health risk as the causative agent of paralytic shellfish poisoning (PSP). Beyond its acute toxicity, STX exerts cascading impacts on food safety, marine ecosystem integrity, and economic stability, particularly in regions affected by harmful algal blooms (HABs). Moreover, the complex molecular structure of STX—tricyclic skeleton and biguanide group—and its diverse analogs (more than 50 derivatives) have made it the focus of research on natural toxins. In this review, we traced the discovery history, chemical structure, molecular biosynthesis, biological enrichment mechanisms, and toxicological actions of STX. Moreover, we highlighted recent advancements in the potential for detection and treatment strategies of STX. By integrating multidisciplinary insights, this review aims to provide a holistic understanding of STX and to guide future research directions for its prevention, management, and potential applications. Full article

In the past two decades, Shiga toxin-producing Escherichia coli (STEC) has been responsible for multiple large-scale outbreaks worldwide, affecting thousands of individuals. While surveillance systems in developed countries such as the United States, the United Kingdom, Europe, Australia, Japan, and Canada are well-established, data on STEC prevalence in developing nations remain sparse, partly due to the absence of well-structured molecular diagnostic networks or surveillance systems. This review analyzed 250 studies published between 2014 and 2024 across 39 developing countries in Africa, Asia, Latin America, and the Caribbean, yielding 8986 STEC isolates. Detailed serogroup and serotype data were available for 55.9% of these, with O111, O157, and O26 being most common in humans. In animals, O157:H7 was most frequent, while food isolates mirrored global trends with O157 and O111 dominance. Notably, O145, a serogroup frequently reported in the U.S. and Europe, was absent from the ‘’Top Seven’’ serogroups. Shiga toxin subtypes stx1a and stx2a were most prevalent in human cases. In animal isolates, stx2e was the most prevalent subtype, while stx2c was most commonly found in food samples. We recommend establishing reference laboratories in these regions to improve data quality, strengthen monitoring efforts, and reduce the burden of STEC infections globally. Full article