Search Results (265)

Green seeds of Coffea arabica represent a rich source of bioactive compounds. This study aimed to compare the butanol-soluble (CA-BU) and the ethyl acetate-soluble (CA-EtAc) fractions in terms of their phytochemical composition and biological activity. As a first step, the fractions were analyzed by Fourier-transform infrared spectroscopy (FT-IR) and high-performance liquid chromatography coupled with mass spectrometry (HPLC–MS) in order to investigate the major constituents. Subsequently, CA-BU and CA-EtAc were evaluated for antioxidant effect, antimicrobial activity, antiproliferative properties, effects on the mitochondrial function, and on the chorioallantoic membrane. The CA-EtAc fraction was enriched in chlorogenic acids and catechins and showed superior antioxidant activity, whereas CA-BU displayed a broader profile of semi-polar polyphenols, conferring moderate antimicrobial effects and stronger antiproliferative activity in MCF-7 human breast adenocarcinoma cells, although with limited selectivity compared with HaCaT non-tumorigenic cells. Respirometric analysis demonstrated that CA-BU selectively inhibited mitochondrial oxidative phosphorylation Complex I (OXPHOS CI), without affecting Complex II (CII) or basal respiration, indicating a specific mitochondria-targeted mechanism. Both fractions were non-irritant and well tolerated in the chorioallantoic membrane (CAM) assay; CA-BU reduced vascular density. These findings demonstrate a clear mechanistic differentiation between the fractions, highlighting the decisive role of solvent polarity in obtaining extracts with distinct and targeted biological effects. Full article

Mitochondrial dysfunction is a hallmark of aging and age-related physical decline in people living with HIV (PLWH) who experience accelerated aging. This pilot study investigated the relationships between platelet mitochondrial function, physical performance, and body composition in older, sedentary PLWH compared with older, sedentary HIV-negative controls. Platelets have the potential to act as minimally invasive and easily accessible biomarkers for systemic mitochondrial bioenergetics and may serve as a practical biomarker in aging-related research. We analyzed correlations between mitochondrial parameters, protein levels, and measures of physical performance and body composition in a cohort of predominantly African American men (n = 7 PLWH, n = 7 controls). Body composition was assessed using dual-energy X-ray absorptiometry (DXA), and exercise capacity through VO₂ peak and strength tests. Platelet mitochondrial bioenergetic parameters were measured by oxygen consumption rates (OCR) and extracellular acidification rates (ECAR). Key mitochondrial proteins SIRT3, COXII, DRP1, and OPA1 were evaluated by Western blotting. The PLWH and HIV-negative control groups were similar in age and cardiorespiratory fitness. In PLWH, basal OCR and ATP-linked respiration showed strong positive correlations with VO₂ peak (r = 0.874, p < 0.05 and r = 0.862, p < 0.05, respectively) and negative correlations with BMI (r = −0.856, p < 0.05 and r = −0.849, p < 0.05, respectively). SIRT3 emerged as a potential key player, demonstrating strong positive correlations with basal OCR (r = 0.804, p < 0.05), ATP-linked respiration (r = 0.787, p < 0.05), and VO₂ peak (r = 0.970, p < 0.001), and negative correlations with BMI (r = −0.830, p < 0.05) and fat mass (r = −0.827, p < 0.05) in PLWH. Analyses focused on within-group associations in PLWH because bioenergetic measures were obtained using different Seahorse platforms in PLWH and controls, precluding valid direct quantitative comparisons between groups. Our findings provide evidence for significant associations between platelet mitochondrial bioenergetics, specific mitochondrial proteins (particularly SIRT3), and key physical attributes in older, sedentary PLWH. These preliminary findings suggest that platelets may serve as minimally invasive biomarkers of systemic mitochondrial health, contribute to our understanding of mitochondrial function in HIV-associated accelerated aging, and inform future interventions to enhance mitochondrial function and improve health outcomes in this vulnerable population. However, results should be interpreted cautiously given the small sample size and exploratory design and should be considered hypothesis-generating rather than definitive. Larger, demographically more diverse studies that include HIV-negative controls are needed to validate these associations and determine their clinical relevance. Full article

Garrya flavescens S. Wats. (GF) has been traditionally used to treat gastrointestinal spasms, yet its bioactivity within the central nervous system remains unexplored. This study aimed to characterize the bioactive constituents of GF and evaluate its anti-inflammatory and metabolic regulatory effects in lipopolysaccharide-activated microglia. Phytochemical profiling using LC-HRMS and HPLC identified rutin as a primary bioactive component, present at an exceptionally high concentration (9309 μg/g). In BV-2 microglial and RAW 264.7 cells, GF treatment significantly suppressed the expression of pro-inflammatory cytokines and mediators in a dose-dependent manner. Mechanistic studies revealed that GF specifically modulated the ERK signaling pathway. Furthermore, Seahorse XF analysis demonstrated that GF restored mitochondrial homeostasis by reducing basal respiration and proton leak while significantly enhancing spare respiratory capacity. Finally, conditioned medium from GF-treated microglia improved the viability of N2A neuronal cells. These findings highlight GF as a potent botanical source with significant neuroprotective potential, offering a promising candidate for functional food or nutraceutical applications targeting neuroinflammatory disorders. Full article

Objective: This study aims to investigate the protective effect of Shengmai San (SMS) against high-glucose (HG)-induced injury in neonatal rat ventricular myocytes (NRVMs) and to elucidate the underlying pharmacological molecular mechanisms. We hypothesize that SMS ameliorates HG-induced calcium homeostasis imbalance in NRVMs by improving mitochondrial energy metabolism disorder, and this protective effect is associated with the downregulation of oxidized and phosphorylated CaMKII expression to inhibit CaMKII signaling pathway overactivation. Herein, we verify this hypothesis by assessing mitochondrial function, calcium transients, sarcoplasmic reticulum (SR) calcium handling and CaMKII phosphorylation levels in NRVMs. Methods: First, ultra-high performance liquid chromatography–high resolution mass spectrometry was used to identify the chemical components of SMS to clarify its material basis. Primary NRVMs were then cultured under low-glucose (LG) or HG conditions, with 2% SMS-medicated serum (SMS-MS) as the experimental intervention, and NAC (ROS scavenger) and KN93 (CaMKII inhibitor) as positive controls. Following intervention, we sequentially detected key indicators corresponding to the proposed pathological pathway: intracellular reactive oxygen species (ROS) levels (oxidative stress), mitochondrial ROS, mitochondrial function indices including oxygen consumption rate (OCR) (energy metabolism), calcium transients and diastolic intracellular free calcium concentration (global calcium homeostasis), sarcoplasmic reticulum (SR) calcium leak (calcium handling disorder), and, finally, the phosphorylation, oxidation levels of CaMKII and RyR2 phosphorylation (Ser2814) (p-RyR2) (key regulatory pathway) via Western blot to systematically elucidate the mechanistic link between SMS intervention and HG-induced NRVM injury. Results: Quantitative analysis revealed that high-glucose (HG) induction significantly reduced calcium transient amplitude and prolonged the decay time constant (tau) in NRVMs at 72 h (p < 0.01 vs. LG), with these parameters normalizing by 120 h—an effect indicative of a compensatory adaptive response. The 2%SMS-MS markedly ameliorated HG-induced calcium transient abnormalities at 72 h (p < 0.01 vs. HG). Additionally, 2%SMS-MS significantly enhanced mitochondrial basal oxygen consumption rate, spare respiratory capacity, ATP production, and maximal respiration in HG-exposed NRVMs (p < 0.01 vs. HG). SMS also significantly reduced intracellular reactive oxygen species (ROS) levels (p < 0.01 vs. HG), mitochondrial ROS levels (p < 0.01 vs. HG), diastolic intracellular free calcium concentration (p < 0.01 vs. HG), and SR calcium leak (p < 0.05 vs. HG). Western blot analysis revealed that 2%SMS-MS intervention effectively downregulated the expression of oxidized CaMKII (Ox-CaMKII) (p < 0.01 vs. HG), phosphorylated CaMKII (p-CaMKII) (p < 0.01 vs. HG), and RyR2 phosphorylation (Ser2814) (p < 0.05 vs. HG), which may be the potential mechanism in maintaining calcium homeostasis in HG-induced NRVMs. Conclusions: This study suggests that SMS enhances mitochondrial energy metabolism and exerts a protective effect against high-glucose-induced calcium homeostasis imbalance in NRVMs, which supports our proposed hypothesis. Its potential mechanism indicates that the protective effects of SMS are associated with its ability to downregulate the expression of oxidized and phosphorylated CaMKII. These findings highlight SMS as a potential therapeutic candidate for alleviating HG-related myocardial injury and provide evidence for its application in the prevention of early diabetic cardiomyopathy. Full article

Soil physicochemical and biochemical properties are fundamental to soil processes and ecosystem functioning in forest environments, yet their responses to dominant tree species in humid montane regions remain largely ununderstood. This study examined the effects of three widespread broadleaf species—Quercus pontica, Quercus petraea, and Fagus orientalis—on soil physical, chemical, and biochemical properties in natural forests in the Eastern Black Sea region, where these species play key ecological roles in structuring forest composition and biogeochemical processes. A total of 15 soil samples (5 per forest type) were collected under comparable climatic and geological conditions and analyzed for particle-size distribution, pH, electrical conductivity (EC), soil organic carbon, and key microbial activity indicators. Significant differences in soil properties were detected among forest types. Soils under Q. pontica were characterized by the lowest silt content and pH, but the highest sand content, soil organic carbon, microbial biomass carbon (C_mic), and microbial respiration. In contrast, soils under Q. petraea exhibited the highest clay content and pH, whereas F. orientalis soils showed lower sand content, EC, soil organic carbon, microbial biomass nitrogen (N_mic), and basal respiration. Multivariate analyses revealed that soil texture, pH, and C_mic are key factors driving soil differentiation across forest types. These patterns indicate that species-specific litter inputs and belowground processes regulate soil biochemical functioning by altering resource availability and habitat conditions. Crucially, this study sheds light on the soil-forming responses of these ecologically dominant species and their impacts on carbon cycle pathways and microbial dynamics at the regional scale. Overall, the study shows that tree species identity is a critical factor influencing soil function, with significant consequences for forest management, carbon sequestration strategies, and ecosystem resilience to changing environmental conditions. Full article

This study aimed to evaluate the effects of adding sludge generated in water treatment plants on the composting of sewage sludge, urban organic waste, and agroindustrial waste. Four treatments were conducted with different proportions of water treatment plant sludge (WTS). Four treatments were conducted with 0%, 10%, 20%, and 30% proportions of WTS. The different proportions allowed for the evaluation of the effects of WTS addition on composting. The study was carried out in composting reactors. Kinetic models were applied to study the degradation of organic matter. Physicochemical and microbiological parameters were analyzed. During the process, temperature variation and basal respiration exhibited similar patterns. Principal component analysis showed that the 30WTS (32.2% water treatment sludge) treatment presented higher values of cation exchange capacity (CEC)/total organic carbon (TOC) ratio (3.83), and germination index (94.35%), and lower values of TOC (23.67%) and C/N (carbon/nitrogen) ratio (14.45). The composts produced in all treatments complied with Brazilian regulations for the environmental and agronomic quality of organic composts. It was concluded that the inclusion of up to 30% of WTS in composting did not negatively affect the composting process and did not compromise the environmental or agronomic quality of the final organic composts. Full article

Background: The pathophysiological mechanisms of Abdominal Aortic Aneurysm (AAA) are not elucidated. Alterations in mitochondrial function, such as a reduction in oxidative phosphorylation (OXPHOS), have been observed at genome level and functionally in vascular smooth muscle cells. Metformin reduces AAA development and growth in diabetic patients, but the precise mechanisms are not known. In this paper we aim to demonstrate the feasibility of measuring mitochondrial functional capacity ex vivo in intact murine aneurysmal tissue and confirm a decrease in OXPHOS, and to determine if the protective effect of metformin on AAA is mediated by mitochondrial function. Methods: AAA was induced in ApoE KO mice by administration of angII (1000 ng/kg/min) through osmotic minipumps. Metformin was administered in drinking water at a dose of 100 mg/kg/day. The abdominal aorta was isolated in situ and mitochondrial functional capacity was analyzed ex vivo in whole permeabilized tissue by high-resolution respirometry. Results: Mitochondrial respiration was successfully measured ex vivo in whole aneurysmal tissue. Mitochondrial function was impaired in angII-treated mice, with decreased fold change in Complex I and Complex I+II oxygen consumption, relative to basal levels. Complex II oxygen consumption was also decreased in angII-treated mice. Rescue treatment of mice with metformin did not affect or restore mitochondrial function. Conclusions: Mitochondrial function can be evaluated in murine whole aneurysmal tissue, providing a method for a physiological approach to the study of mitochondrial function in AAA. Mitochondrial function is impaired in AAA. However, rescue treatment with metformin is not sufficient to recover mitochondrial function and seems not to be the mechanism behind prevention of aneurysm. Full article

Chronic kidney disease (CKD) involves uremic toxin-driven tubular injury and systemic vascular dysfunction, in which mitochondrial impairment and apoptotic cell loss contribute to progressive tissue deterioration. Accordingly, a targeted EV platform is required to enable efficient miRNA delivery to the toxin-stressed tubular–endothelial compartment. Based on our previous study showing that melatonin restores miR-4516 levels under CKD-related stress, we directly loaded miR-4516 into engineered extracellular vesicles (EVs) to evaluate its effects on mitochondrial function and cell survival. Here, we engineered EVs with a G3-C12/RGD surface modification and established a miR-4516 loading strategy to enhance delivery to kidney proximal tubule cells and vascular endothelial cells. miR-4516 loading increased EV-associated miR-4516 levels without major changes in particle size distribution, and EV identity was supported by CD9 and CD81 expression. Confocal microscopy and flow cytometry demonstrated increased cellular uptake of miR-4516-loaded G3-C12/RGD-EVs compared with control EVs in TH1 proximal tubule cells and HUVECs. Under indoxyl sulfate stress, engineered EV treatment restored intracellular miR-4516 and improved mitochondrial function, as indicated by recovery of respiratory Complex I and Complex IV activities and improved Seahorse bioenergetic parameters (OCR/ECAR, basal and maximal respiration, ATP-linked respiration, and spare respiratory capacity). Annexin V staining further indicated reduced toxin-induced apoptosis. In an adenine diet-induced CKD mouse model, intravenous administration of miR-4516-loaded G3-C12/RGD-EVs improved urinary albumin-to-creatinine ratio (UACR), blood urea nitrogen (BUN), and serum creatinine. These findings indicate that miR-4516-loaded, targeting-engineered EVs may mitigate uremic toxin-associated mitochondrial dysfunction and renal impairment in CKD. Full article

Background/Objectives: Kidneys from marginal donors are more susceptible to ischemia–reperfusion injury (IRI). To diminish the deleterious effects of IRI, mitochondria should be preserved optimally between donation and transplantation. It is unknown which macronutrients are imperative to support mitochondrial function during normothermic preservation. The aim of this study is to investigate the effect of different macronutrient compositions on mitochondrial function during prolonged normothermic incubation of precision-cut kidney slices. Methods: Both porcine (n = 8) and human (n = 5) kidneys were studied. After warm ischemia and cold preservation, precision-cut kidney slices (PCKS) were made and incubated in different incubation media under normothermic conditions. PCKS were incubated with all different combinations of glucose, glutamine and/or fatty acids. At zero, 24 and 48 h, mitochondrial function and metabolite levels were assessed. Results: After 48 h of incubation, ATP levels were significantly higher with glucose and glutamine or fatty acids and glutamine, compared to incubation without nutrients (p = 0.001 and p = 0.003, respectively). Also, mitochondrial oxygen fluxes were higher with all nutrients compared to no nutrients (ADP stimulated basal respiration p = 0.040; proton leak p = 0.001 and maximal respiration p = 0.039). Conclusions: The addition of multiple macronutrients during prolonged normothermic incubation increases the mitochondrial function of PCKS. These data pave the way for optimising the nutritional support for ex vivo perfusion of marginal kidneys. Full article

Airborne particulate matter with a diameter of <10 μm (PM₁₀) can damage the corneal epithelium by inducing oxidative stress, disrupting the NRF2 antioxidant pathway, and triggering epithelial barrier dysfunction and inflammation. However, the role of mitochondria in mediating PM₁₀-induced damage remains unexplored. This study investigated the impact of PM₁₀ on mitochondrial homeostasis in both immortalized human corneal epithelial cells (HCE-2) and the mouse corneal epithelium, as well as the protective effects of SKQ1. For in vivo assessment, female C57BL/6 mice were exposed to either control air or PM₁₀ (±SKQ1) in a whole-body exposure chamber for 2 weeks (3 h/day, 5 days/week, with weekends off). In vitro, HCE-2 cells were exposed to 100 μg/mL PM₁₀ (±SKQ1) for 24 h, and mitochondrial function and morphology were evaluated. In vitro, PM₁₀ significantly impaired mitochondrial function by reducing basal, maximal, and ATP-linked respiration; reserve capacity; and coupling efficiency compared to the control and SKQ1 groups. PM₁₀ also downregulated mitofusin1 (MFN1) and optic atrophy1 (OPA1) and upregulated dynamin-related protein1 (DRP1) and mitochondrial fission protein1 (FIS1) in HCE-2 cells. In addition, PM₁₀ exposure significantly decreased the mitochondrial membrane potential; mitochondrial DNA copy number; and cytochrome c oxidase subunit 4 isoform 1 (COX4i1), mitochondrial transcription factor A (TFAM), and peroxisome proliferator-activated receptor gamma coactivator 1 alpha (PGC-1α) levels. SKQ1 pre-treatment significantly attenuated these effects. In vivo, PM₁₀ exposure significantly decreased the levels of MFN1, TFAM, COX4i1, and superoxide dismutase (SOD2), whereas SKQ1 treatment significantly reversed these effects. Overall, these findings demonstrate that PM₁₀ exposure induces mitochondrial fragmentation, disrupts mitochondrial biogenesis and quality control, and reduces mitochondrial respiration, resulting in mitochondrial dysfunction. SKQ1 effectively reversed these changes, suggesting its potential as a therapeutic strategy to protect corneal epithelial cells from PM₁₀-induced mitochondrial damage. Full article

Background: Normobaric acute hypoxia models are widely applied to assess tolerance to acute hypoxic stress. Highland barley is a cereal crop originating from and traditionally cultivated in high-altitude regions; however, the dose–response relationship underlying its effects on hypoxia tolerance remains unclear. Methods: Male ICR mice were randomly allocated to five groups (n = 8 per group) and fed an AIN-93M basal diet or experimental diets supplemented with 20%, 40%, 60%, or 80% highland barley for 13 weeks. Hypoxia survival time was evaluated using a normobaric asphyxial hypoxia model, in which oxygen is progressively depleted in a sealed chamber by continuous respiration with carbon dioxide absorbed by soda lime. Hematological parameters, indices of oxidative stress and energy metabolism, and gut microbiota composition were also assessed. Results: Compared with the control group, dietary supplementation with 20% highland barley was associated with a longer hypoxia survival time (mean difference: 9.49 min; 95% CI: −2.05 to 21.02), whereas the 80% group exhibited the shortest survival time (approximately 40.6 min). In the 20% group, red blood cell count and hemoglobin concentration increased by 41.6% and 42.1%, respectively. ATP content and superoxide dismutase activity in brain tissue increased by 33.2% and 28.4%, respectively, with similar trends observed in heart tissue. In addition, gut microbiota α-diversity was increased in the 20% highland barley group, and distinct separation of microbial community structures was observed among groups receiving different supplementation levels. Conclusions: Overall, the data suggest that moderate dietary supplementation with highland barley (20%) is associated with a favorable physiological and microbiota profile under normobaric asphyxial hypoxic challenge, suggesting the presence of a potentially effective intake range for highland barley-based nutritional intervention. Full article

Riparian forests play a critical role in protecting soil and water resources and maintaining ecosystem stability. In this study, we evaluated the response of soil chemical and microbial attributes to different stages of riparian forest restoration in the protection zone of the Itaipu Reservoir (Brazil). Soil samples were collected during summer and winter from sites representing four restoration stages (initial, 3, 19, and 30 years), as well as from an adjacent agricultural field and a native forest used as reference systems. We assessed soil chemical properties, microbial biomass carbon, basal respiration, enzymatic activities, and the soil microbial community structure using 16S rRNA gene sequencing. Principal component analysis (PCA) revealed a clear restoration gradient, with older restored sites progressively converging toward the native forest condition. Soil chemical properties showed gradual recovery along the restoration trajectory, with increases in soil organic carbon, cation exchange capacity, and base saturation. In contrast, the availability of P, K, Ca, and Mg declined at early restoration stages and increased with restoration age. Microbial biomass carbon increased by approximately 60% from early restoration to native forest conditions, while metabolic quotients (qCO₂) decreased, indicating greater microbial efficiency and reduced metabolic stress. Enzyme activities related to C, P, and S cycling increased by 1.5- to 3-fold with restoration age. Sequencing analyses indicated a progressive convergence of microbial community composition toward that of the native forest, driven by shifts in relative abundance and the enrichment of forest-associated taxa, such as Verrucomicrobia and Acidobacteria, at advanced restoration stages. Overall, long-term riparian forest restoration promoted substantial recovery of soil chemical fertility and microbial community structure and functioning, reinforcing the role of soil microbiota as a sensitive indicator of ecosystem resilience and restoration success. Full article

Superabsorbent polymers (SAPs) are materials that can absorb and retain water solutions with a mass of several hundred times greater than their own. This work aimed to synthesise and evaluate the effects of highly absorbent starch phosphate-g-poly(acrylic acid) copolymers on the microbiological activity of soils previously used for agriculture. The biopolymers studied were obtained by thermal and chemical oxidation of starch phosphates and copolymerized with potassium salts of acrylic acid. Basic physicochemical parameters were determined in the applied soil. Following SAP application, the basal respiration rate was measured at 22 °C with a constant soil moisture content of 60% WHC. The incubation time in constant temperature and moisture conditions was 78 days. After this period, their microbiological activity (microbial and organic phosphorus fractions) was assessed, thereby enabling the determination of the direction of change in the soil environment. The addition of SAP increases the soil’s water-holding capacity and respiration. The SP-g-PAA polymers serve as slow-release sources of potassium and phosphorus ions. These elements were bound to the polymer network by ionic and covalent bonds. Analysis of the results shows that within two weeks, 47–80% of the starch hydrogel undergoes microbial degradation. No differences were found in the content of labile forms of phosphorus in soils with SAP additions compared to soils without polymer additions. The use of modified starch reduces the consumption of vinyl monomers, while the resulting product is characterised by high absorbency and low water content, which reduces the amount of energy needed to obtain the finished product, thus contributing to sustainable development. Full article

The intensive cultivation of greenhouse tomatoes generates massive quantities of vegetative residues often laden with potentially complex pesticide contaminants, posing a dual challenge of waste management and environmental toxicity. This study investigated the biological feasibility and system tolerance of valorizing these hazardous residues through vermicomposting with Eisenia fetida, using mixtures of cattle manure and tomato residues (TR) at varying ratios (0–60%) over a 45-day incubation period. The process was monitored through physicochemical parameters (pH, EC, C/N ratio) and sensitive biological indicators (Basal Respiration and Microbial Biomass Carbon). While TR inclusion rates exceeding 30% induced acute inhibitory effects (100% mortality within 5 days) due to acute toxicity, mixtures containing up to 30% were successfully processed. The biological monitoring revealed a distinct “biphasic response”: an initial “metabolic lag phase” (days 0–15) driven by chemical stress, followed by a robust “biological recovery” where microbial activity surged significantly after day 30. Correlation analyses confirmed that this recovery was mechanically linked to the acidification of the substrate, as indicated by strong negative correlations between pH and biological activity (r_s = −0.70). Ultimately, vermicomposting significantly reduced Electrical Conductivity (EC) and lowered the C/N ratio below 15 in all viable treatments, confirming the stabilization of waste into an agronomically mature product. The results demonstrate that the earthworm gut functions as an effective bioreactor, facilitating biological stabilization and the mitigation of toxicity in pesticide-laden biomass. This study concludes that vermicomposting is a robust strategy for converting toxic agro-wastes into a stabilized organic amendment, provided that the residue load is managed within the identified physiological tolerance threshold of 30%. Full article

This study evaluates the technical feasibility of decentralized vermicompost production by smallholder farmers within a structured rural development program. Conducted under the KOP-TEYAP initiative in Kırşehir Province, Türkiye, the research assesses whether farmers can consistently produce vermicompost that meets international quality standards following a participatory training and infrastructure support model. Fourteen farmers, selected through a merit-based process from 232 trainees, were provided with standardized production units. The produced vermicompost was analyzed for critical chemical parameters (pH, EC, organic matter, C:N ratio, K, Cu, Zn) and biological indicators (basal CO₂ respiration, microbial biomass carbon) and benchmarked against regulations from the EU, France, Germany, Austria, Canada, India, and Türkiye. Results indicated that the majority of farmer-produced samples successfully met the critical thresholds for chemical quality and safety. Furthermore, biological maturity was confirmed by low basal respiration levels and high microbial biomass across the samples. These findings demonstrate that structured farmer training combined with standardized low-cost infrastructure enables smallholders to reliably produce high-quality vermicompost, validating this model as an effective agroecological strategy for rural development. Full article