Search Results (889)

Ochratoxin A (OTA), a prevalent food contaminant, is closely linked to the development of various cancers, including clear cell renal cell carcinoma (ccRCC). However, the potential mechanisms remain to be explored. In this study, we employed network toxicology, machine learning, and molecular docking techniques to systematically investigate the potential molecular mechanisms underlying OTA-associated ccRCC. We normalized transcriptional data from two Gene Expression Omnibus (GEO) datasets and analyzed it using differential expression analysis and weighted gene co-expression network analysis (WGCNA), identifying 3224 ccRCC-associated target genes. These were intersected with 232 predicted OTA target genes, yielding a total of 56 overlapping targets. The results of Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) analyses indicated that these targets were primarily enriched in critical biological processes, including extracellular matrix remodeling, immune microenvironment regulation, signaling pathway transduction, cellular metabolism, and protein homeostasis. Machine learning analysis identified “glmBoost + RF” (a sequential combination of feature selection and classifier) as the optimal model, from which nine key genes were extracted. SHapley Additive exPlanations (SHAP) analysis revealed five core genes (IGFBP3, ITGA5, PYGL, SLC22A8, LTB4R), with IGFBP3 and ITGA5 serving as the principal driver genes of the model. Validation of the model’s diagnostic efficacy and single-cell transcriptome analysis indicated that the core genes exhibited significant differential expression patterns, cell-type-specific expression characteristics, and high independent diagnostic efficacy. Molecular docking analyses predicted stable interactions between OTA and the core target proteins. These findings suggest potential molecular links between OTA exposure and ccRCC, providing a foundation for hypothesis generation and future experimental validation. Full article

Ochratoxin A (OTA) is a highly toxic mycotoxin commonly detected in food and agricultural products, requiring sensitive analytical methods for reliable monitoring. Herein, we report an ultrasensitive turn-on electrochemical aptasensor for OTA detection based on a target-induced displacement of an aptamer–complementary DNA (cDNA) duplex assembled on an electrochemically reduced graphene oxide (ERGO)-modified glassy carbon electrode (GCE). In the absence of OTA, a methylene blue (MB)-labeled aptamer hybridized with cDNA is immobilized on the ERGO surface via π–π stacking interactions, forming a rigid duplex that suppresses electron transfer and yields a low electrochemical signal. Upon OTA binding, the aptamer undergoes a conformational transition into a G-quadruplex structure, leading to dissociation of the cDNA strand. This target-induced folding brings the MB redox tag into close proximity to the ERGO surface, markedly accelerating electron transfer and enhancing the cathodic reduction current of MB, thereby producing a pronounced signal-on response in square-wave voltammetry (SWV). The ERGO-modified electrode provides a conductive and stable interface without chemical linkers. Under optimized conditions, the aptasensor shows a linear response to OTA from 10 fM to 100 pM with an ultralow LOD of 0.67 fM, together with high selectivity, good reproducibility, and satisfactory stability. This work demonstrates a simple and effective turn-on aptasensing strategy for sensitive electrochemical detection of OTA. Full article

Human organoids and organ-on-chip/microphysiological systems (OoC/MPS) are increasingly used as new-approach methodologies for biotoxin assessment. They retain human-relevant tissue organization and enable interpretable analysis of exposure geometry, barrier transport, perfusion, and (when needed) multi-organ coupling. In this review, we synthesize primary evidence across major toxin classes, including bacterial enterotoxins (e.g., cholera toxin, heat-stable enterotoxins, Shiga toxins), mycotoxins (e.g., aflatoxin B1, ochratoxin A, deoxynivalenol), and algal/cyanobacterial toxins (e.g., saxitoxin, domoic acid, microcystins, biliatresone). We emphasize studies that clearly define toxin identity and exposure context and that demonstrate mechanism-critical model competencies under assay conditions. We highlight decision-informative functional endpoints that align with the dominant pathophysiology. These include cystic fibrosis transmembrane conductance regulator (CFTR)-dependent secretion in human enteroids/colonoids, transporter-linked proximal tubular injury in kidney MPS, gut–kidney axis injury from Shiga toxin-producing E. coli in microfluidic systems, and multi-electrode array (MEA) network readouts in human 3D neural tissues. We then summarize best practices that improve cross-study comparability. These include reporting delivered versus nominal exposure, assessing recovery/mass balance and device/material interactions, applying proportional biological qualification (polarity, transporter/enzymatic competence, functional stability), defining a minimal comparable endpoint core, and preserving QIVIVE readiness in reporting. Finally, we outline near-term priorities for the field, including chronic low-dose and mixture designs, harmonized reference panels and acceptance criteria, and fit-for-purpose escalation to coupled OoC/MPS only when perfusion or organ–organ coupling is expected to change the interpretation. Full article

Cecina de León is a traditional Spanish dry-cured beef product whose surface, as in other similar meat products, becomes heavily colonised by fungi during ripening, raising concerns related to possible mycotoxin contamination. This study aimed to characterise the mycobiota associated with cecina and its production environment, with particular emphasis on mycotoxigenic Penicillium species. Seventy-eight cecina samples and 26 air samples were collected from meat-processing plants and local markets in the province of León (Spain) and analysed for fungal counts, water activity and pH. A total of 101 mould isolates and 16 yeasts were recovered, with Penicillium accounting for 88% of all moulds. Sixteen Penicillium species were identified using a polyphasic approach integrating macro- and micromorphological analysis, extrolite production, molecular markers (BenA, CaM and ITS), and MALDI-TOF MS. Mycotoxin screening by HPTLC and HPLC-PDA targeted cyclopiazonic acid, ochratoxin A, patulin, citrinin, griseofulvin and mycophenolic acid, revealing that 51% of the Penicillium isolates were mycotoxin producers, mainly P. commune. The proposed polyphasic strategy, including MALDI-TOF MS as a rapid complementary tool, offers a practical framework for the surveillance of fungal communities and mycotoxin risk in meat-processing environments. Full article

The objective of this study was to evaluate the preharvest application of γ-aminobutyric acid (GABA), melatonin (MT), and oxalic acid (OA), at different concentrations and application frequencies, on the physicochemical and microbiological quality of dried figs (cv. Calabacita) at commercial harvest and after 3 and 6 months of refrigerated storage. A further aim was to determine their impact on fungal populations and mycotoxin production. The results showed that untreated dried figs had a higher frequency of Aspergillus welwitschiae, A. tubingensis, and Aspergillus section Flavi, whereas elicitor-treated figs exhibited a lower incidence of toxigenic fungi. A. welwitschiae was the main ochratoxin A (OTA)-associated species detected, although the proportion of OTA-positive figs was lower in elicitor-treated samples than in the control. Aflatoxins (AFs) were detected only sporadically in 2 mM OA treatments, consistent with the limited activity of A. flavus at low storage temperatures. Conversely, Penicillium spp. were widespread but were associated with citrinin (CIT) production only under 2 mM OA treatments. Among the Alternaria toxins, alternariol (AOH) was detected solely in dried figs treated with 1 mM OA. Notably, all investigated mycotoxins were below the limit of detection (<LOD) in dried figs treated with 0.5 mM MT. Moderate elicitor concentrations (e.g., 0.5 mM MT and 50 mM GABA) and multiple preharvest applications generally provided the best balance between fungal suppression and fruit quality, significantly reducing Aspergillus spp. occurrence without promoting the growth of undesirable species. Overall, elicitor treatments decreased the incidence of toxigenic fungi, most likely through direct antifungal effects in senescent dried fruit rather than by inducing host defences. The combined use of preharvest elicitors with appropriate drying and storage conditions is a promising strategy to control fungal contamination and mycotoxin accumulation in dried figs while maintaining quality from preharvest storage. Further research is needed to optimise elicitor concentrations and application timing. Full article

Ochratoxin A (OTA), a secondary metabolite produced by Penicillium and Aspergillus species, contaminates food and feed globally, posing serious threats to both livestock and human health. Among current detoxification strategies, probiotic-based degradation of OTA has emerged as a key research focus. This study aimed to isolate safe probiotic strains with high OTA-detoxifying efficacy to support their potential application in feed and food industries. A total of 57 bacterial strains were isolated from environmental samples, including soil, moldy feed, and animal feces. Among these, a novel strain identified as Bacillus amyloliquefaciens MM28 demonstrated strong OTA-degrading activity, removing 86.31% of OTA (0.4 µg/mL) within 48 h. Whole-genome analysis indicated that B. amyloliquefaciens MM28 harbors functional genes related to glucose metabolism, membrane transport, and properties associated with antibacterial, antioxidant, and immunomodulatory activities, suggesting multiple beneficial traits. In a 28-day chronic exposure study, mice were administered B. amyloliquefaciens MM28 via gavage (1 × 10⁸ CFU/mL). Results showed that both female and male mice in the MM28 group exhibited higher body weight and improved growth performance compared to the PBS control group. Furthermore, intestinal morphology was enhanced in the MM28 group, as indicated by greater villus length and villus-length-to-crypt-depth ratio. The expression of proinflammatory cytokines was also reduced in the treated animals. Moreover, analysis of gut microbiota composition revealed that MM28 supplementation led to an increased abundance of Bacteroides and Desulfovibrio, alongside a reduction in Lachnospira and Oscillospira. In conclusion, this study demonstrates that Bacillus amyloliquefaciens MM28 is a safe and efficient strain capable of degrading OTA. These findings highlight its promising potential as a biological detoxifying agent in food and feed industries. Full article

Climate change is intensifying ecological instability in agricultural ecosystems, altering pest distribution and increasing the risk of fungal contamination and mycotoxin accumulation in crops. Among emerging threats, the invasive brown marmorated stink bug Halyomorpha halys has rapidly expanded across Europe, including Croatia. The species is polyphagous, and in the ripening of most plant crops, it accounts for a significant population and causes extensive plant damage. This study investigates the capacity of H. halys to disseminate mycotoxigenic fungi, with an emphasis on aflatoxin B₁ (AFB₁) and ochratoxin A (OTA) producers. One hundred adult H. halys individuals were collected from maize ears in Osijek, Croatia, in August 2023. Fungal propagules washed from their surfaces were inoculated on PDA and MEA media. A total of 212 pure fungal cultures were obtained, of which 202 belonged to the genera Aspergillus and Penicillium. Molecular identification using β-tubulin (BenA) gene sequencing revealed six Aspergillus and six Penicillium species, forming two well-supported phylogenetic clades. Aspergillus parasiticus was the most dominant species. Mycotoxin screening confirmed AFB₁ production in 20 isolates, all identified as A. parasiticus, while no OTA-producing strains were detected. The predominance of aflatoxigenic species on H. halys indicates that it may play a previously underrecognized role in the dispersal of mycotoxigenic fungi in agroecosystems. Full article

This study used the QuEChERS method in combination with liquid chromatography–quadrupole-time-of-flight mass spectrometry (LC-Q-TOF/MS) to develop a method for simultaneous detection of 187 pesticides and 10 mycotoxins in Astragalus. The samples were extracted using an acetonitrile–water solution containing 5% formic acid, and the amount of purification materials was optimized through response surface methodology. The results show that 197 compounds exhibit good linear relationships within their respective linear ranges (R² > 0.995). The screening detection limits (SDLs) and the limits of quantification (LOQs) ranged from 0.001 to 0.02 mg/kg and 0.002 to 0.02 mg/kg, respectively. At the spiked levels of 1, 2, and 10 times LOQ, compound recoveries ranged from 61.5% to 118.9%, 67.1% to 119.6%, and 72.0% to 119.3%, respectively, with relative standard deviations (RSDs) all less than 20.0%. The intra-day precision and inter-day precision are less than 10% and 20%, respectively. This method was applied to detect 20 batches of commercially available Astragalus samples. Six compounds (three pesticides and three mycotoxins) were detected; the residues of aflatoxin and ochratoxin A in two batches exceeded the maximum residue limits and required attention. The established method is simple, rapid, and highly sensitive. It is also reproducible and meets the requirements for the accurate quantitative analysis of multiple pesticide residues and mycotoxins in Astragalus. Full article

Aspergillus westerdijkiae is a common pathogenic fungus responsible for postharvest fruit rot in pears, causing substantial economic losses. This fungus also produces ochratoxin A (OTA), which poses serious health risks to humans. During host colonization, fungal pathogens secrete effectors to facilitate invasion. Under host-mimicking culture conditions, transcriptomic analysis of A. westerdijkiae at 24 and 72 h post-inoculation (hpi), combined with signal peptide prediction, identified 272 and 214 up-regulated secreted protein-encoding genes, respectively. Among these, a carboxylesterase gene, AwCES, was found to be significantly up-regulated. Compared to the wild-type strain, deletion of AwCES resulted in reduced conidial production and germination rate. Further studies revealed that the deletion mutant showed significantly attenuated virulence on pear fruit. Moreover, the loss of AwCES impaired fungal adaptation to stress environments. Collectively, these findings demonstrate that AwCES plays a critical role in the growth, development, and pathogenicity of A. westerdijkiae. Full article

Ochratoxin A (OTA) is a globally distributed mycotoxin that poses serious threats to food safety and human health due to its nephrotoxic, hepatotoxic, and carcinogenic properties. Previous enzymatic detoxification strategies for OTA have been constrained by low degradation efficiency or poor soluble expression of highly active enzymes. In this study, a bacterial strain with strong OTA-degrading activity was isolated and identified as Acinetobacter kookii AK4, which degraded 95.44% of 1 μg/mL OTA within 6 h. The predominant OTA-degrading activity was derived from intracellular enzymes. Through genome mining and experimental validation, gene2102 was identified as encoding an amidohydrolase. The enzyme was designated AMH2102 and was heterologously expressed in Escherichia coli. Codon optimization combined with fusion of an N-terminal SUMO tag increased the soluble expression of AMH2102 by 14.81-fold, enabling complete (100%) OTA degradation within 3 min. Overall, this study achieved the identification of an efficient OTA-degrading strain and enzyme and explored strategies for improving enzyme expression, yielding effective outcomes that provide useful references for future studies on strain mining and enzyme engineering. Full article

Meat and meat products are integral components of various diets and provide many macro- and micronutrients. However, concerns over their potential adverse health effects remain pressing. This study employed a semi-quantitative Risk–Benefit Assessment (RBA) methodology to compare both beneficial and adverse health impacts of various meat products, i.e., fermented (Salame and Chouriço), dry-cured (Presunto), and heat-treated (Fiambre), with unprocessed grilled pork meat as a reference. Nutritional composition and microbiological and toxicological hazards were assessed using data acquired from national and international databases and the literature. In the end, a heatmap approach was used to summarize and compare product profiles. While grilled pork offered the most favorable profile, processed products showed high levels of sodium, nitrites, and contaminants, such as polycyclic aromatic hydrocarbons and ochratoxin A. Notably, Salame exhibited the most concerning risk profile, including high levels of histamine and ochratoxin A, whereas Fiambre, despite the high nitrite content, showed the lowest microbial and toxicological risks. These findings highlight significant variability in health-related impacts among meat products, caused mainly by processing technologies. The results can improve dietary guidance and regulations and encourage innovations, especially by indicating the potential of using engineered fermentation techniques and novel additives for improved meat products. Full article

This study was aimed at producing nanocellulose from sugarcane bagasse and durian rind residues for applications to determine adsorption capacity against mycotoxin in poultry diets. Durian rind-derived nanocellulose exhibited finer fiber (12–21 nm diameter and 197–350 nm length) and higher yield (42.1%) than bagasse-derived nanocellulose (18–36 nm diameter and 82–169 nm length), with FTIR confirming purer cellulose I/II structures. The in vitro test adsorption capacity against ochratoxin (OTA) was determined at an incubation time of 180 min to establish working conditions. It was found that the working conditions of bagasse-derived nanocellulose and durian rind-derived nanocellulose were 33 mg/mL and 36.5 mg/mL, respectively. Subsequently, using these working conditions, adsorption capacity was determined via an in vitro digestibility test. Bagasse-derived nanocellulose exhibited an adsorption capacity against OTA of 35.59%, while durian rind-derived nanocellulose achieved an OTA adsorption rate of 39.53% at a contact time of 3 h. Naturally contaminated poultry feeds collected from nine farms in Chiang Mai, Thailand, indicated that both types of nanocelluloses achieved minimum–maximum OTA adsorption rates of 42–43%, aflatoxin B1 (AFB1) at 29–30%, and fumonisin B1 (FB1) at 21–23% across the nine farms’ mean values. These findings suggest that nanocellulose derived from sugarcane bagasse and durian rind has potential as a sustainable biosorbent for improving mycotoxin management in poultry production. Full article

This study evaluated the occurrence of ochratoxin A (OTA) in Turkish coffee and its potential health implications under current consumption patterns by analyzing 65 ground and roasted Turkish coffee samples collected across Türkiye. OTA contamination was detected in 53 samples (82%). Based on the mean OTA concentration, the Estimated Daily Intake (EDI) was calculated as 0.1403 ng/kg body weight/day, and health risk characterization was performed using the Margin of Exposure (MOE) approach in accordance with the European Food Safety Authority (EFSA) recommendations for chronic exposure assessment. MOE calculations enabled a refined characterization of health risks under realistic (0.5 cup/day), average (1 cup/day), and high (3 cups/day) consumption scenarios. The MOE values for carcinogenic (neoplastic) effects ranged from 34,450 to 206,847, all exceeding the EFSA reference threshold of 10,000 and indicating a low level of concern for carcinogenic risk associated with Turkish coffee consumption. For non-carcinogenic (non-neoplastic) kidney effects, MOE values ranged from 11,238 to 67,475 across the different consumption scenarios, all exceeding the EFSA reference threshold of 200, indicating a low level of concern for the general population. In conclusion, the findings demonstrate that Turkish coffee consumption does not pose an OTA-related carcinogenic or non-neoplastic health risk for the general population under current consumption patterns. Nevertheless, considering the widespread consumption of Turkish coffee, continued monitoring and strict implementation of control measures throughout the production chain remain advisable to ensure long-term consumer safety. Full article

Background/Objectives: Mycotoxin contamination in grain-derived foods is still a major food safety concern; thus, innovative mitigation approaches need to be continuously developed. This study investigated the influence of bilberry (Vaccinium myrtillus L.) incorporated into a food matrix on ochratoxin A (OTA)-induced cellular responses using a dietary-relevant in vitro intestinal model. Methods: Four bread types were prepared: control (C), OTA-contaminated (OTA), bilberry-enriched (VM), and OTA + VM (OTA-VM). Simulated intestinal digests of these breads were applied to differentiated Caco-2 cells for 24 h. Apoptotic and necrotic cell populations, as well as reactive oxygen species (ROS) levels, were quantified by flow cytometry, while RT-qPCR assessed the expression of 10 genes related to mitochondrial function, oxidative stress response, and intestinal barrier integrity. Results: Exposure to OTA resulted in increased cytotoxicity, reflected by a higher proportion of necrotic cells (5.11 ± 0.35%), and elevated ROS levels compared with control cells. Co-exposure to bilberry-enriched digests was associated with attenuation of apoptotic responses, a reduced proportion of necrotic cells (2.16 ± 0.61%) and a 16% decrease in ROS levels. Gene expression profiles in the VM group were comparable to control, whereas OTA exposure led to downregulation of several genes related to oxidative stress response and intestinal barrier integrity (e.g., CLDN2, OCLN, SLC7A11). In the OTA-VM group, a partial recovery of gene expression was observed. Conclusions: These findings suggest that bilberry incorporation into a food matrix may modulate OTA-induced cellular stress responses by attenuating oxidative imbalance and supporting the expression of genes associated with antioxidant defense and epithelial barrier integrity. Bilberries may therefore represent a promising functional ingredient for influencing intestinal cellular responses to dietary mycotoxin exposure. Full article

Bongkrekic acid (BKA), a highly lethal toxin, has been implicated in frequent poisoning incidents in recent years, posing a serious threat to global food safety and creating an urgent need for rapid and sensitive detection methods. This review provides a systematic analysis of the entire BKA detection technologies, covering sample pretreatment techniques, instrumental analysis, immunoassays, and biosensing methods. It assesses the merits of key methods and also explores the strategic cross-application of detection paradigms developed for analogous toxins. This review delivers a comprehensive and critical evaluation of BKA detection technologies. First, it discusses sample pretreatment strategies, notably solid-phase extraction (SPE) and QuEChERS. Subsequently, it analyzes the principles, performance, and applications of core detection methods, including high-performance liquid chromatography–tandem mass spectrometry (HPLC-MS/MS), high-resolution mass spectrometry (HRMS), time-resolved fluorescence immunoassay (TRFIA), dual-mode immunosensors and nanomaterial-based sensors. Instrumental methods (e.g., HRMS) offer unmatched sensitivity [with a limit of detection (LOD) as low as 0.01 μg/kg], yet remain costly and laboratory-dependent. Immunoassay and biosensor approaches (TRFIA and dual-mode sensors) enable rapid on-site detection with high sensitivity (ng/mL to pg/mL), though challenges in stability and specificity remain. Looking forward, the development of next-generation BKA detection could be accelerated by cross-applying cutting-edge strategies proven for toxins—such as Fumonisin B1 (FB1), Ochratoxin A (OTA), and Aflatoxin B1 (AFB1)—including nanobody technology, CRISPR-Cas-mediated signal amplification, and multimodal integrated platforms. To translate this potential into practical tools, future research should prioritize the synthesis of high-specificity recognition elements, innovative signal amplification strategies, and integrated portable devices, aiming to establish end-to-end biosensing systems capable of on-site rapid detection through multitechnology integration. Full article