The integration of micro- and macroethical perspectives remains a challenge in engineering ethics education (EEE). Fiction-based EEE has been advocated as a pedagogical approach that can help students in this regard. However, how students actually use fiction to engage with micro- and macroethics remains underexplored. Therefore, this case study aimed to (1) identify how students' engagement with fictions connected to micro and macro perspectives on engineering and technology ethics, (2) examine how students realized pedagogical potentials of fiction-based pedagogy in this regard, and (3) explore how the outcomes related to the pedagogical framing and activities of the course. The selected case was an elective, fiction-based EEE course, during which the written course work of students, teacher and student interviews, course materials, and data from workshop observations were collected for qualitative analysis. Findings show that several expected pedagogical potentials of fiction-based pedagogy were realized as students reflected on what guides human behavior and on the societal impact of technology, both within and beyond the fictions. However, students' reflections rarely connected to their future professional role. These findings could be related to characteristics of the employed pedagogy and fictions, but also to students' prior knowledge and expectations regarding professional engineering practice. We conclude that students may need targeted support towards connecting fictions with self and profession, and that fiction-based EEE may benefit from recognizing and responding to students' pre-existing expectations on social responsibility and ethical agency in engineering.
To address the food safety risks caused by carbendazim (CBZ) residues, a high-efficiency self-powered photoelectrochemical (PEC) sensing platform was constructed for sensitive detection of CBZ. We present a PEC sensing interface based on a Pt nanoparticles-engineered ZIF-8 core encapsulated by a covalent organic framework (COF) shell (Pt NPs-ZIF-8@COF). Distinct Pt integration pathways were evaluated, revealing that in situ encapsulation of Pt NPs during ZIF-8 crystallization (Pt NPs-ZIF-8) mitigates particle aggregation and outperforms post-synthetic decoration of Pt NPs onto post-synthetic ZIF-8 (Pt NPs/ZIF-8). The COF overlayer further tailors the interfacial microenvironment by establishing a continuous charge-transport network, which together accelerate photogenerated charge separation and interfacial electron-transfer processes. Benefiting from these synergistic features, the resulting PEC sensor enables quantitative CBZ determination over a wide linear range from 1.0 × 10- 8 to 1.0 × 10- 2 µg/mL, with an ultralow detection limit of 3.3 × 10- 9 µg/mL. This work underscores a modular nanoparticles-in-MOF plus COF encapsulation strategy for building high-performance PEC biosensing interfaces and offers a promising route toward rapid pesticide-residue monitoring.
Polyvinyl chloride (PVC) is one of the most widely used polymers on a global scale. However, its ability to withstand natural degradation presents substantial environmental concerns. It is imperative to identify biological solutions for PVC degradation. Consequently, the present investigation examined the potential of the haloalkane dehalogenase LinB from Sphingomonas paucimobilis bacteria to degrade PVC by cleaving its strong C-Cl bonds. Molecular docking and molecular dynamics simulations were employed to determine the small PVC fragments at the molecular level, utilizing advanced computational techniques. The analysis suggested that the PVC fragment maintains a stable configuration throughout and bonds well to the enzyme's active site. Subsequently, the stable configuration of PVC along with the nearby residues was subjected to a detailed QM/MM calculation. The results indicated that the carbon-chlorine bond is weakened and approaches a pre-dissociation state. The vibrational analysis suggested weakening of the C-Cl bond together with the development of C-O interaction features, consistent with a reactive configuration. This study also involved the cap-domain mutations (V134L, A135L, and V134L/A135L) of LinB to improve PVC dechlorination by stabilizing catalytically favorable substrate configurations rather than improving binding affinity. Molecular docking results with binding scores of - 5.9 to - 6.0 kcal-1, MD simulations (300 ns), and MM-GBSA analysis results showed the optimized mutants. Mutant-2 maintained the consistent proximity between Asp108 and the chlorine-bearing carbon atom. These results provide new quantum-level insights into the interaction of LinB with PVC-derived fragments and useful information for enzyme engineering that is better for clearing PVC plastic waste.
Monosex and sterile fish populations are increasingly recognized as essential tools for enhancing aquaculture productivity while safeguarding ecosystems from genetic introgression and uncontrolled reproduction. Conventional approaches such as hormonal manipulation and triploidy have demonstrated utility but face persistent limitations in efficiency, welfare, and regulatory acceptance. Recent breakthroughs in CRISPR-based genome editing and germ cell transplantation now enable heritable, scalable, and biocontained control of sex and fertility, offering transformative potential for sustainable aquaculture. This paper synthesizes advances across finfish and crustaceans, critically evaluates technical and regulatory bottlenecks, and highlights convergent solutions integrating automation, artificial intelligence, and governance frameworks. By positioning reproductive programming as a cornerstone of climate-resilient and sustainable aquatic food systems, we propose a roadmap toward commercial adoption that balances biological precision, ecological safety, and societal acceptance.
Mesenchymal stromal cells (MSCs) have demonstrated significant potential for various clinical applications; however, several challenges limit their therapeutic efficacy. These include donor variability, poor homing and survival after transplantation, reduced proliferative and differentiation potential during in vitro expansion, and the inflammatory microenvironment at the injury site. This review summarizes current strategies used to enhance the therapeutic potential of MSCs and discusses approaches to overcome these limitations. It also highlights key challenges and gaps in the existing literature that require further investigation. In addition, this structured review provides an overview of current research trends involving different MSC sources, disease models, preconditioning strategies, emerging gene modification approaches, and other methods to improve MSC function. Finally, regulatory considerations and the roles of global and regional regulatory authorities in ensuring the safety and quality of modified MSC-based therapies are briefly discussed.
Interface-driven hotspot-engineered flexible optical sensors are reported having been prepared by immobilizing anisotropic nanostars (NS) on polypropylene (PP) films for in situ multiplexed detection of pesticide and synthetic colourant residues using a chemometric-assisted surface-enhanced Raman spectroscopy (SERS) strategy. Highly anisotropic nanostars were synthesized via a facile one-pot chemical reduction approach, with controlled nanostar morphologies facilitating tunable plasmonic activity. Detailed structural, optical and chemical compositional investigation confirmed effective integration of nanostars into PP fibrous matrices. The SERS performance was systematically modulated by varying the nanostar spike geometry, achieving a maximum enhancement factor of 9.6 × 107. Furthermore, finite-element electromagnetic (EM) simulations revealed strong localization of the electromagnetic field around nanostar tips, with optimal enhancement at 785 nm excitation, thereby confirming the dominance of tip-induced EM enhancement. The surface interactions leading to dense nanostar hotspots on fibrous PP mats have been well elucidated, considering the hydrophobicity-assisted nanostar confined assembly. The developed sensors demonstrated superior signal reproducibility, stability, and detection limits reaching femtomolar levels for methylene blue and nanomolar levels for thiram and carmoisine. The selective identification of pesticides and synthetic colourant residues was achieved through multivariate chemometric analysis, which demonstrated clear spectral discrimination. The real-time detection of thiram and carmoisine in commercial fruit juice samples was achieved with excellent specificity and recoveries of 92 to 109%, which illustrated the practical applicability of PP/NS sensor for food safety monitoring applications.
Hydrogels with high strength, stiffness, and toughness under full hydration are essential for load-transfer applications such as artificial tendons, ligaments, and soft robotics. Yet achieving such performance remains difficult because conventional polymer networks are intrinsically soft and transfer stress inefficiently. Here, a covalent interfacial anchoring (CIA) strategy is introduced to enable high-strength, high-stiffness hydrogel microfibers under full hydration by chemically anchoring carbon nanotubes (CNTs) and graphene oxide (GO) within poly(vinyl alcohol) networks. In this network, CNTs contribute to axial load transfer, whereas GO forms glutaraldehyde-mediated PVA-GO acetal linkages that suppress nanofiller mobility during deformation and promote efficient stress transfer. The resulting hydrogel fibers achieve tensile strength of 132 MPa, modulus of 1.1 GPa and toughness of 25 MJ m-3 under full hydration and maintain ∼88% displacement after 100 tendon-mimetic loading cycles. These findings highlight covalent interfacial anchoring as an effective strategy for engineering strong and stiff hydrogel fibers for tendon-like load-transfer applications.
pH-responsive DNA nanodevices and materials are attracting wide attention owing to their promising applications in medicine and nanotechnology. However, the sustainable and systematic development of molecular pH-switches based on mismatch-guided hybridization of G-quadruplexes and i-motifs lacks a rational framework for their design and analytical analysis. In this study, we formulate and rigorously analyze the principles underlying the operation of such molecular nanomachines. Using the AS1411 aptamer to nucleolin as a model G-quadruplex, we designed 19 pH-switches having a transition pH from 5.2 to 7.4 to validate our framework. Ultimately, we derived empirically grounded guidelines for the engineering of the pH-switches and thermodynamic approaches to predict their properties analytically.
The NLRP3 inflammasome plays an important role in the innate immune system that mediates caspase-1 activation and the subsequent secretion of the pro inflammatory cytokines IL-1β and IL-18 in response to microbial infections and cellular stress. Aberrant activation of the NLRP3 inflammasome has been implicated in a range of inflammatory and metabolic disorders, including cryopyrin-associated periodic syndromes (CAPS), Alzheimer's disease, type 2 diabetes, and atherosclerosis. NLRP3 activation can be triggered by diverse stimuli and involves multiple cellular events, such as ionic flux, mitochondrial dysfunction, reactive oxygen species (ROS) generation, and lysosomal damage. A critical regulator of NLRP3 activation is NEK7, a serine/threonine kinase also known for its role in mitotic spindle formation and cytokinesis. Structurally, NEK7 exhibits a partially assembled regulatory spine (R-spine), consistent with an inactive kinase state. However, a mutant form of NEK7 engineered to stabilize the R-spine retains catalytic activity and has been crystallized in complex with compound 51- an ATP-competitive, small-molecule chemical inhibitor that targets both NEK2 and NEK7. Interestingly, wild-type NEK7 crystallized in both apo form and with compound 51 showed variable R-spine conformations, while NEK7 displayed a uniform, partially stacked R-spine configuration. Despite the structural similarity in compound 51 binding between NEK2 and NEK7, subtle differences in ligand orientation reveal potential avenues for designing isoform-selective inhibitors. In this review, we comprehensively examine the structural domains and subcellular localizations of NLRP3 and NEK7, and highlight how their interaction promotes inflammasome assembly. I also discuss recent insights from cryo-electron microscopy (cryo-EM), X-ray crystallography, and molecular modeling studies that illuminate the conformational dynamics of the NLRP3-NEK7 complex. Additionally, we explore the crosstalk between the STING pathway and inflammasome signaling, and address key unanswered questions that may guide future research into therapeutic modulation of these essential protein-protein interactions.
The escalating crisis of antimicrobial resistance (AMR) necessitates the urgent development of alternatives to traditional antibiotics. Bacteriophage (phage) therapy, which utilizes viruses that specifically infect and lyse bacteria, has re-emerged as a promising therapeutic strategy. This review comprehensively examines the current landscape, beginning with the modern, genome-based classification of phages and detailing their key therapeutic advantages, including high specificity, self-amplification, and biofilm-penetrating capability. We explore advanced biotechnological applications such as genetic engineering of phages, the use of phage-derived proteins, and synergistic phage-antibiotic combinations (PAS). The translational workflow from phage sourcing and biobanking to characterization, formulation, and clinical delivery is critically analyzed, alongside major therapeutic areas like chronic wounds and pulmonary infections. Despite promising clinical evidence from compassionate use and trials, significant scientific, regulatory, and commercial hurdles remain. The integration of synthetic biology and artificial intelligence is poised to overcome these challenges, steering phage therapy toward becoming a precise and adaptable component of the modern antimicrobial arsenal.
Radioactive isotopes, such as Indium-111 (111In), are pivotal for highly sensitive noninvasive imaging and targeted radiotherapy. However, achieving stable in vivo retention remains a critical barrier. Conventional nanocarriers typically rely on surface-bound chelators, exposing the isotopes to biological transchelation and detachment, while potentially altering the carrier's surface properties and pharmacokinetics. To address these limitations, this study evaluates a polyionic complex micelle (PICm) platform designed to encapsulate 111In within a crosslinked core using DOTA chelation. This approach aims to enhance radiochemical stability while preserving the micelle's native, engineered surface characteristics. DOTA-functionalized PICm were fabricated through electrostatic self-assembly of PEG-based polymers and subsequent core crosslinking. The resulting micelles exhibited a highly uniform size (32.6 ± 6.3 nm) and a modestly negative surface charge (- 4.9 ± 1.0 mV). Stability assays demonstrated that under the acidic conditions required for 111In chelation (pH 5.5), the micelles undergo reversible size swelling driven by polymer protonation without aggregating or collapsing, confirming that core crosslinking successfully maintains micellar integrity. In vivo biodistribution in CT26 tumor-bearing mice revealed that 111In-loaded PICm displayed prolonged systemic presence, with tumor accumulation peaking at 2.3 ± 0.4%ID/g at 24 h post-injection via the enhanced permeability and retention effect. Hepatic and splenic uptakes aligned with typical nanoparticle clearance pathways. The core-loading of PICm may offer advantages over surface-chelated nanocarriers. By maintaining structural integrity during radiolabeling and extending systemic circulation in vivo, these results validate the platform's robustness and passive tumor accumulation, highlighting its potential as a versatile vehicle for the delivery of diagnostic radionuclides.
Amorphous PdAg alloy nanoparticles with tunable compositions were synthesized via a simple one-pot strategy. Composition and phase engineering synergistically optimize CO2 electroreduction, with an amorphous Pd86Ag14 alloy catalyst delivering superior CO selectivity and activity over amorphous Pd and crystalline PdAg alloys, providing an effective route to high-performance amorphous alloy catalysts.
Magnetically actuated surgical systems represent a disruptive class of mechatronic devices that offer a promising approach to enhance minimally invasive (MIS) procedures by reducing tissue trauma. This literature systematic review synthesizes 10 years evidence, from 2016 to 2026, on magnetic mechatronics and robotics in hepatopancreatobiliary (HPB), bariatric, gynecologic, urologic, and neurosurgical applications, focusing on design principles, actuator-sensor fusion, and performance compared to non-magnetic systems. Following PRISMA 2020 guidelines, we searched Scopus using specific Boolean strings for magnetic actuation in the surgical field. Eligibility required experimental validation and clinical translation, prioritizing Q1 and Q2 journals. After title, abstract, and full-text screenings, 187 of 68,956 studies were included. Data extraction focused on technical metrics like coupling efficiency and positional precision, alongside clinical outcomes such as safety and invasiveness. In this sense, a narrative synthesis grouped findings by mechanism and specialty. In correlation, key findings highlight coupling models such as dipole-dipole and gradient-driven force transmission. Indeed, performance envelopes showed sub-millimetric accuracy and improved workspaces for retraction and anchoring over conventional tools, though thermal and saturation limits persist. These systems demonstrate potential for multispecialty translation, establishing a mechatronic framework to guide future platform designs. Limitations include prototype heterogeneity and limited large-scale clinical data. This analysis underscores engineering constraints and opportunities for advanced actuator-sensor integration in MIS procedures.
Upcycling CO2 into battery components promises carbon-neutral energy storage, yet most CO2-derived polymers require harsh synthesis conditions and have limited electrochemical integration. This work reports a hybrid polymer electrolyte based on a CO2-derived polyurethane (PCO2), which is synthesized under ambient conditions (room temperature, 1 atm) as a carbon-utilization strategy. A composite electrolyte (GPCO2-2) is further constructed by integrating this PCO2 with a polymer matrix and a deep eutectic solvent component, wherein in situ densification and interfacial reconstruction are established to simultaneously achieve mechanical integrity and electrochemical stability. Benefiting from the abundant coordination sites within GPCO2-2, a competitive coordination mechanism prevents excessive Li+ binding while ensuring continuous ion transport pathways. This unique configuration endows the electrolyte with seamlessly coupled bulk-interface ion transport properties, significantly enhanced Li+ dissociation and migration, and promotes the formation of a dense and stable solid-electrolyte interphase. Consequently, the electrolyte delivers outstanding stability, enabling lithium metal to cycle reliably for over 2500 h. At a voltage of 4.3 V, the Li/GPCO2-2/NCM811 battery achieves 500 stable cycles at 0.5 C. Even under a higher voltage of 4.7 V and at 0.2 C, it still completes 100 stable cycles. Flexible pouch cells assembled with this GPCO2-2 electrolyte show viability with 71.1% capacity retention after 100 cycles. Beyond cycling performance, the electrolyte's robust oxidative stability also endows the cells with enhanced safety features. Overcharge tests indicate that the GPCO2-2 cells maintain stable current at high voltages and a lower temperature rise, enhancing safety and oxidative stability through regulated Li+ transport.
Bio-based fermented products have recently become increasingly important for both human health and sustainability due to the growing interest in healthy sustainable diets. In this study, spirulina was used as a substrate for the cultivation of Lacticaseibacillus rhamnosus 23.2 in a 3 L bioreactor and emphasized the evaluation of the potential health effects of the obtained bioactive products. The antioxidant potential of bioactive products has been extensively studied using assays like DPPH (2,2-diphenyl-1-picrylhydrazyl) and ABTS (2,2'-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid)) radicals, demonstrating its capacity to scavenge free radicals and mitigate oxidative stress. MTT and cell migration (scratch) assays reduced viability of Caco-2 cells and hindered cell migration cancer cells FS compared to unfermented spirulina (unFS) revealed that FS significantly reduced cell viability and migration in a dose-dependent manner, with the strongest effects observed at 5X dilution. In contrast, unFS showed weaker bioactivity. These findings highlight fermentation as a promising biotechnological approach to enhance the functional properties of natural compounds for cancer prevention and treatment. To summarize, the current study revealed that the potential of spirulina biomass to be a suitable media substrate for L. rhamnosus 23.2 strain. These findings will provide awareness toward milestone the health and food sector industrialization of FS products.
A novel ternary composite of zirconium-based supramolecular organic framework/fullerene@strontium-based metal-organic framework (Zr-SOF/C60@Sr-MOF) was synthesized and adopted as an electrode modifier to construct a high-sensitivity electrochemical sensor for ciprofloxacin (CIP) determination. The composition and morphology of the material were confirmed by SEM, TEM, XPS, XRD and FT-IR. Cyclic Voltammetry (CV) and electrochemical impedance spectroscopy (EIS) were used to evaluate the electron-transfer properties and interfacial behavior of the modified electrodes, and square wave voltammetry (SWV) was employed for the sensitive and quantitative detection of the target analyte. The results reveal that the composite inherits the abundant porous structure of Zr-SOF, the expansive specific surface area of Sr-MOF and excellent electron transfer capability of fullerene. The sensor enables accurate CIP detection in the linear range 5.0 nM-1000 µM, with a detection limit of 7.0 nM (S/N = 3), along with satisfactory selectivity, reproducibility and long-term stability. Practical application was validated in lake water, tap water and milk samples, yielding recoveries of 95.3%-101.63%. This work provides new insights into SOF/MOF-based electrochemical sensing and establishes an efficient strategy for rapid on-site detection of CIP antibiotics in environmental and food matrices.
Skeletal muscle plasticity progressively declines from middle age onward, and exercise may partially mitigate this deterioration through microRNA (miRNA)-mediated regulatory networks. However, whether the timing of aerobic exercise initiation during middle age (early vs. late) differentially influences muscle function and miRNA expression profiles remains incompletely understood. Male C57BL/6 mice were assigned to four groups: middle-aged control (MC, 12 months), old-aged control (MC18, 18 months), early exercise (PRE18, 12-15 months), and late exercise (POS18, 15-18 months). Gastrocnemius muscles were collected for miRNA sequencing and subsequent functional analyses. The MC18 group exhibited sarcopenic-like phenotypes, with 176 differentially expressed miRNAs (DEmiRNAs) identified relative to the MC group. Aerobic exercise was associated with attenuation of age-related muscle atrophy, with PRE18 showing comparatively more pronounced effects than POS18. miRNA sequencing identified 21 DEmiRNAs with reversed expression patterns in the PRE18 group, whereas only 2 such miRNAs were identified in the POS18 group. Bioinformatic analysis of predicted targets suggested that early-exercise-induced miRNAs may be involved in suppressing FOXO/ubiquitin-mediated proteolysis and restoring Wnt/cAMP signaling, whereas late-exercise-induced miRNAs appeared to have a limited capacity to restore Wnt signaling. miR-195a-5p_R+1, miR-298-5p_R-2, miR-671-5p_R+1 and miR-150-5p were identified as candidate hub miRNAs based on target enrichments within these pathways. Western blot analyses indicated that PRE18 was associated with elevated expression of Wnt‑related (β‑catenin) and cAMP‑related (CREB1, PGC‑1α) proteins, alongside reduced levels of atrophic markers (FOXO3A, FBXO32, and MuRF1). In contrast, POS18 primarily affected atrophic protein expression, with relatively modest influence on Wnt signaling components. These preliminary findings collectively suggest that the sustained benefits of exercise initiated in early middle age might be partially attributable to exercise‑induced miRNAs dually targeting both regenerative and atrophic pathways. Conversely, late initiation appears to be predominantly linked to anti‑atrophic effects, with less impact on regenerative signaling. Together, these observations provide initial evidence for temporal epigenetic plasticity and suggest broader molecular advantages of earlier intervention, although further mechanistic studies are warranted.
The ability of hydrogels to promote wound healing has been extensively studied. Developing multifunctional hydrogel dressings to address the complex microenvironment of infected wounds remains a significant challenge and focus in current research. Herein, inspired by adhesion chemistry, we constructed a multifunctional composite hydrogel (GelDA/OPL) with excellent adhesion, self-healing properties, injectability, and photothermal antibacterial activity through Schiff base crosslinking between dopamine-modified gelatin (GelDA) and oxidized pullulan (OPL). By varying the concentration of OPL, the mechanical and rheological properties of GelDA/OPL can be appropriately adjusted. Furthermore, by introducing phycocyanin-modified CeO2@PC NPs, we endowed GelDA/OPL with remarkable antioxidant characteristics capable of rapidly scavenging ˙OH, ABTS˙+, and DPPH˙ radicals. The results indicate that the incorporation of catechol groups not only enhanced the adhesive performance of GelDA/OPL/CeO2@PC hydrogels but also imparted exceptional photothermal conversion efficiency under 808 nm laser irradiation, effectively inhibiting Staphylococcus aureus and Escherichia coli. The applicability of GelDA/OPL/CeO2@PC hydrogels in promoting wound healing in vivo was further validated using a full-thickness skin defect infection model in rats. Overall, the prepared GelDA/OPL/CeO2@PC hydrogel represents a promising multifunctional wound dressing that contributes to accelerating the healing process for infected wounds.
Synthesizing zeolites from natural kaolin clay presents a sustainable and cost-effective alternative to traditional synthetic precursors. The different variants of zeolites, characterized by their unique Si/Al ratios and pore structures, can significantly affect their selectivity and adsorption behaviors when interacting with various hydrocarbon gases. This study seeks to explore and explain how hydrocarbon gases (CH₄, C₂H₆, C₃H₈) engage with zeolite frameworks on a molecular level. To achieve this, zeolite variants were synthesized by etching kaolin with different concentrations of sulfuric acid (20%, 40%, and 60%). The resulting zeolites were tailored to have adjustable Si/Al ratios and porosity, which were then characterized through BET, SEM-EDX, and XRD techniques. The molecular interactions between the zeolite variants and hydrocarbons were investigated using MEP and NBO analyses to connect the electronic properties with the variations in etching. The findings revealed that the LUMO-HOMO energy gap ranges from 2.396 to 2.944 eV, showing a correlation with the zeolite framework and Si/Al ratio. Moreover, the adsorption behavior exhibited differing selectivity, stabilization energy, and reactivity with the three hydrocarbons, highlighting trends that depend on the framework. Changes in bond lengths within the zeolite-hydrocarbon complexes, spanning 3.107 to 4.196 Å, suggest that adsorption occurs in a less confined environment. This could enhance diffusion and improve the kinetics of adsorption and desorption, offering promising implications for gas purification in pressure swing adsorption (PSA) systems. The computational analysis and molecular modelling of the zeolite-hydrocarbon gases were carried out using the Gaussian 16.0 and Gausview 6.0 visualization software packages, to present an understanding of how hydrocarbon gases are adsorbed by various commercial and lab-created zeolites via molecular electrostatic potential (MEP) and natural bond orbital (NBO) charge analysis. The DFT was adopted to optimize all zeolite-hydrocarbon complexes described by the HOMO-LUMO energy gap of the hydrocarbon complexes, which were calculated at the (B3LYP-D3/6-311 + + G(d,p)) method. This approach provides an insight into the hydrocarbon gas selectivity of the zeolite surface at the atomic level. The differences in reactivity and electronic stability (energy gap) can be utilized in pressure swing adsorption (PSA) processes to enhance selectivity in refineries and other real-world applications.
Enterocytozoon bieneusi is a widespread intracellular fungal pathogen that infects a variety of hosts. To investigate its prevalence and genetic diversity in captive Tarim red deer in southern Xinjiang, we collected 630 fresh fecal samples from 10 farms between 2020 and 2021. PCR targeting the ITS gene revealed an overall infection rate of 23.5% (148/630), with 8 out of 10 farms testing positive. Infection rates varied significantly by farm (12.2%-83.9%, P < 0.01) and age group (P < 0.01), peaking at 53.9% in 12-24-month-old deer and dropping to 0.6% in 6-12-month-old individuals. Sequencing identified 22 genotypes: 7 known and 15 novel (XJRD1-XJRD15), with BEB6 (54.7%) and D (21.6%) being the most prevalent. Phylogenetic analysis categorized 16 genotypes into zoonotic Group 1 and 6 into host-adapted Group 2, with novel genotypes forming distinct subclades. Tarim red deer serve as a crucial reservoir for E. bieneusi diversity and pose zoonotic risks, highlighting the need for targeted One Health interventions. Our study revealed a high prevalence of infection and significant genetic diversity of E. bieneusi in captive Tarim red deer, including the identification of novel genotypes. Several of the detected genotypes have zoonotic potential, suggesting that these deer may serve as reservoirs.