Search Results (157)

Fusarium sporotrichioides strain M-1-1, originally deposited as Fusarium solani IFO 9955 in 1974 and later moved to NBRC, is known for producing T-2 toxin. In addition to NRRL 3299, which was used in the United States to study T-2 toxin biosynthesis, NBRC 9955 has been extensively used for trichothecene research in Japan. To facilitate and accurately document studies on trichothecene biosynthesis using NBRC 9955, its phylogenetic classification and trichothecene metabolite profiles were determined. As anticipated, NBRC 9955 was classified as F. sporotrichioides, which exhibited a more distant phylogenetic relationship to other strains within the same species. Time-course TLC analyses demonstrated the accumulation of various deacetylated trichothecenes in yeast extract-rich liquid media during the late growth stages. Conversely, an increase in 3-O-acetylation of T-2 toxin was observed at late stages when cultivated in micronutrient-poor synthetic liquid medium. Northern blot analysis revealed that Tri8 expression halted in cultures with the synthetic medium, which accounts for the growth stage-dependent 3-O-acetylation observed. On a brown rice flour solid medium, the fungal strain produced mixtures of T-2 toxin, neosolaniol, HT-2 toxin, and their 3-O-acetyl derivatives. These results highlight the risk of underestimating the levels of toxic trichothecene metabolites when using the standard contamination monitoring protocols. Full article

This study evaluated 65 commercially available pet feed samples, including 33 cat feeds and 32 dog feeds (dry and wet formulations), for the presence of organic contaminants. These included mycotoxins, pesticides, pharmaceutical residues/veterinary drugs, and plant-based bioactive compounds. A suspect screening strategy was employed using QuEChERS extraction followed by LC-LTQ/Orbitrap HRMS analysis. A total of 29 compounds were tentatively identified within 186 detections. In total, 76.9% of the samples were contaminated with mycotoxins. Aflatoxins (B1, B2, G1, and G2), T2 toxins, and HT2 toxins were dominant, with Aflatoxin B1 occurring in 33.8% of the samples and exhibiting a higher prevalence in dry feeds than in wet feeds. Pesticides were present in 72.0% of the dry formulations, including aclonifen and pirimiphos-methyl, but were present in only 11% of the wet formulations. Plant-based bioactive compounds, including phytoestrogens, were identified in 51% of the samples, highlighting toxicologically relevant candidates that merit prioritization for targeted confirmation, particularly in cat feeds. Pharmaceuticals were found in 23.8% of dry feeds (sparfloxacin and fumagillin). Overall, the HRMS-based, standard-free suspect screening workflow provides an early-warning overview of multi-class co-occurrence patterns in complex pet feed matrices and supports the prioritization of candidates for subsequent confirmatory analysis. Full article

This study aimed to conduct a first evaluation of maternal exposure to mycotoxins during pregnancy in Brazil through quantification of biomarkers in liver and serum samples from stillborn and neonates autopsied in the Clinical Hospital of Ribeirão Preto, state of São Paulo. Liver tissue (n = 43) and serum (n = 38) samples were collected from 43 patients and analyzed for biomarkers of aflatoxins (AFs), ochratoxin A (OTA), fumonisins (FBs), zearalenone (ZEN), deoxynivalenol (DON), T-2 and HT-2 toxins by ultra-performance liquid chromatography coupled to tandem mass spectrometry. In total, 9 samples of liver (20.9%) and 33 serum (86.8%) had quantifiable levels of mycotoxins. In liver samples, aflatoxin M₁ (AFM₁) was quantified in two samples (median level: 0.38 ng/g), while four samples had OTA residues (median: 0.31 ng/g) and one contained ZEN (3.6 ng/g). Compared with liver tissue, serum samples had higher occurrence rates of mycotoxins, particularly AFM₁, OTA and ZEN. Nineteen serum samples (50%) contained 2–4 types of mycotoxins, indicating an effective transplacental transfer of major mycotoxins during pregnancy. Median levels of AFM₁, OTA, FB₁, ZEN, DON, T-2 and HT-2 toxins in serum samples were 0.48, 3.39, 30.6, 10.53, 5.71, 2.85 and 10.84 ng/mL, respectively. The most frequent cause of death was extreme prematurity (33% of cases), followed by preterm premature rupture of membranes (16% of cases) and morphological abnormalities (42% of cases). Results of this trial suggest potential associations between dietary mycotoxins and congenital anomalies. Further research should clarify the transplacental transfer of mycotoxins and their association with toxic effects during human prenatal development. Full article

This study aimed to verify the simultaneous presence of mycotoxins in feed intended for food-producing animals. A validated liquid chromatography–mass spectrometry analytical method was used for the determination and quantification of aflatoxins (AFB1, AFB2, AFG1, AFG2), deoxynivalenol, ochratoxin A, fumonisins, T-2 and HT-2 toxins, and zearalenone. The correlation coefficient indicated a good fit for all analytes, ranging from 0.991 to 0.999, while the mean recoveries were between 76 and 108%. The occurrence of one or more mycotoxins was detected in 42% of all feed samples investigated, at concentrations ranging between 0.0030 and 0.042 mg/kg for AFB1 and 0.16 and 0.95 and 0.016 and 1.5 mg/kg for deoxynivalenol and zearalenone, respectively. The sum of T-2 and HT-2 toxins ranged from 0.011 to 0.088 mg/kg, while the sum of fumonisins was between 0.010 and 14 mg/kg. Twenty-four positive samples (28%) showed the co-presence of ZEA and/or DON with FB1 and FB2, six of which were also contaminated with T-2 and HT-2 toxins. The need for continuous monitoring is particularly emphasized to ensure the health of both animals and humans. Full article

T-2 toxin and HT-2 toxin are commonly found in agricultural products and animal feed, posing serious effects to both humans and animals. This study employed combination index (CI) modeling and metabolomics to assess the combined cytotoxic effects of T-2 and HT-2 on four porcine cell types: intestinal porcine epithelial cells (IPEC-J2), porcine Leydig cells (PLCs), porcine ear fibroblasts (PEFs), and porcine hepatocytes (PHs). Cell viability assays revealed a dose-dependent reduction in viability across all cell lines, with relative sensitivities in the order: IPEC-J2 > PLCs > PEFs > PHs. Synergistic cytotoxicity was observed at low concentrations, while antagonistic interactions emerged at higher doses. Untargeted metabolomic profiling identified consistent and significant metabolic perturbations in four different porcine cell lines under co-exposure conditions. Notably, combined treatment with T-2 and HT-2 resulted in a uniform downregulation of LysoPC (22:6), LysoPC (20:5), and LysoPC (20:4), implicating disruption of membrane phospholipid integrity. Additionally, glycerophospholipid metabolism was the most significantly affected pathway across all cell lines. Ether lipid metabolism was markedly altered in PLCs and PEFs, whereas PHs displayed a unique metabolic response characterized by dysregulation of tryptophan metabolism. This study identified markers of synergistic toxicity and common alterations in metabolic pathways across four homologous porcine cell types under the combined exposure to T-2 and HT-2 toxins. These findings enhance the current understanding of the molecular mechanisms underlying mycotoxin-induced the synergistic toxicity. Full article

Mycotoxins represent a group of highly toxic secondary metabolites produced by diverse fungal pathogens. Mycotoxin contaminations frequently occur in foods and feed and pose significant risks to human and animal health due to their carcinogenic, mutagenic, and immunosuppressive properties. Notably, deoxynivalenol, zearalenone, fumonisins (mainly including fumonisins B1, B2, and FB3), aflatoxin B1 (AFB1), and T-2/HT-2 toxins are the major mycotoxin contaminants in foods and feed. Undoubtedly, exposure to these mycotoxins can disrupt gut health, particularly damaging the intestinal epithelium in humans and animals. In this review, we summarized the detrimental effects caused by these mycotoxins on the intestinal health of humans and animals. The fundamental molecular mechanisms, which cover the induction of inflammatory reaction and immune dysfunction, the breakdown of the intestinal barrier, the triggering of oxidative stress, and the intestinal microbiota imbalance, were explored. These signaling pathways, such as MAPK, Akt/mTOR, TNF, TGF-β, Wnt/β-catenin, PKA, NF-kB, NLRP3, AHR, TLR2, TLR4, IRE1/XBP1, Nrf2, and MLCK pathways, are implicated. The abnormal expression of micro-RNA also plays a critical role. Finally, we anticipate that this review can offer new perspectives and theoretical foundations for controlling intestinal health issues caused by mycotoxin contamination and promote the development of prevention and control products. Full article

Several species of Fusarium are reported to infect inflorescences of high-THC-containing cannabis (Cannabis sativa L.) plants grown in greenhouses in Canada. These include F. graminearum, F. sporotrichiodes, F. proliferatum, and, to a lesser extent, F. oxysporum and F. solani. The greatest concern surrounding the infection of cannabis by these Fusarium species, which cause symptoms of bud rot, is the potential for the accumulation of mycotoxins that may go undetected. In the present study, both naturally infected and artificially infected inflorescence tissues were tested for the presence of fungal-derived toxins using HPLC-MS/MS analysis. Naturally infected cannabis tissues were confirmed to be infected by both F. avenaceum and F. graminearum using PCR. Pure cultures of these two species and F. sporotrichiodes were inoculated onto detached inflorescences of two cannabis genotypes, and after 7 days, they were dried and assayed for mycotoxin presence. In these assays, all Fusarium species grew prolifically over the tissue surface. Tissues infected by F. graminearum contained 3-acetyl DON, DON, and zearalenone in the ranges of 0.13–0.40, 1.18–1.91, and 31.8 to 56.2 μg/g, respectively, depending on the cannabis genotype. In F. sporotrichiodes-infected samples, HT2 and T2 mycotoxins were present at 13.9 and 10.9 μg/g in one genotype and were lower in the other. In F. avenaceum-inoculated tissues, the mycotoxins enniatin A, enniatin A1, enniatin B, and enniatin B1 were produced at varying concentrations, depending on the isolate and cannabis genotype. Unexpectedly, these tissues also contained detectable levels of 3-acetyl DON, DON, and zearalenone, which was attributed to apre-existing natural infection by F. graminearum that was confirmed by RT-qPCR. Beauvericin was detected in tissues infected by F. avenaceum and F. sporotrichiodes, but not by F. graminearum. Naturally infected, dried inflorescences from which F. avenaceum was recovered contained beauvericin, enniatin A1, enniatin B, and enniatin B1 as expected. Uninoculated cannabis inflorescences were free of mycotoxins except for culmorin at 0.348 μg/g, reflecting pre-existing infection by F. graminearum. The mycotoxin levels were markedly different between the two cannabis genotypes, despite comparable mycelial colonization. Tall fescue plants growing in the vicinity of the greenhouse were shown to harbor F. avenaceum and F. graminearum, suggesting a likely external source of inoculum. Isolates of both species from tall fescue produced mycotoxins when inoculated onto cannabis inflorescences. These findings demonstrate that infection by F. graminearum and F. avenaceum, either from artificial inoculation or natural inoculum originating from tall fescue plants, can lead to mycotoxin accumulation in cannabis inflorescences. However, extensive mycelial colonization following prolonged incubation of infected tissues under high humidity conditions is required. Inoculations with Penicillium citrinum and Aspergillus ochraceus under these conditions produced no detectable mycotoxins. The mycotoxins alternariol and tentoxin were detected in several inflorescence samples, likely as a result of natural infection by Alternaria spp. Fusarium avenaceum is reported to infect cannabis inflorescences for the first time and produces mycotoxins in diseased tissues. Full article

Maize is one of the most important agricultural crops that has been cultivated in the Republic of Croatia for centuries. Fusarium mycotoxins as secondary metabolites of molds that naturally contaminate maize crops can have negative effects on human and animal health, but also on economic aspects. The aim of this study was to monitor the trend of natural occurrence of Fusarium mycotoxins in maize crops from Croatia during a period of 10 years, from 2014 to 2024. A total of 1285 maize samples were analyzed for the contents of zearalenone (ZEN), deoxynivalenol (DON), fumonisins B (FUM) and T-2/HT-2 toxin (T-2/HT-2) using validated ELISA methods. A high occurrence of the analyzed mycotoxins was found in all years examined, e.g., for ZEN 15–64%, DON 47–95%, FUM 60–96% and T-2/HT-2 24–89%. In addition, their concentrations ranged from 3.2–10,990 μg/kg, 18–25,000 μg/kg, 29–18,180 μg/kg and 9–595 μg/kg, respectively. The mycotoxin concentrations were found to differ from year to year, with an increasing trend observed for FUM and T-2/HT-2 in terms of concentrations and an increasing trend in the occurrence of all mycotoxins, with the exception of T-2/HT-2. Full article

This study investigates the presence of mycotoxins in green coffee-based dietary supplements to ensure their safety, given the potential risks of contamination and the growing interest in them among consumers. A sample treatment based on a salting-out assisted liquid–liquid extraction (SALLE) followed by one-step solid-phase extraction (SPE) was selected for the extraction and clean-up of 15 mycotoxins followed by ultra-high performance chromatography–tandem mass spectrometry detection (UHPLC-MS/MS). The target mycotoxins included aflatoxins (AFG1, AFG2, AFB1, AFB2), Alternaria toxins (AOH, AME, TEN), ochratoxin A (OTA), fumonisins (FB1, FB2), zearalenone (ZEN), trichothecenes (T-2, HT-2), enniatin B1 (ENNB1), and beauvericin (BEA). The proposed method was successfully characterized, obtaining high recoveries, a satisfactory precision, and low detection limits. Subsequently, the method was applied for the analysis of 16 commercial food supplements. The analysis revealed the presence of mycotoxins in all samples investigated with Fusarium mycotoxins as the most prevalent. The dietary exposure and risk characterization revealed a low level of risk, except for AFs where chronic exposure in adults may lead to potential health concerns. Full article

Mycotoxin contamination in maize poses significant food and feed safety risks, particularly in regions with variable climatic conditions like Serbia. This study investigated the occurrence of regulated mycotoxins in maize harvested across the Republic of Serbia from 2021 to 2023, emphasizing the impact of climatic factors. A total of 548 samples of unprocessed maize grains were analysed for the presence of key mycotoxins, including aflatoxins, ochratoxin A, zearalenone, deoxynivalenol, fumonisins, and trichothecenes type A (T-2 and HT-2 toxins), using validated analytical methods. The results revealed high contamination frequencies, with aflatoxins and fumonisins being the most prevalent. The results revealed substantial temporal variability and frequent co-contamination of mycotoxins. Aflatoxin B₁ (AFB₁) was the most concerning contaminant, with 73.2% of the samples in 2022 exceeding the European regulatory limit for human consumption (5 µg/kg) for un processed maize grains, reaching peak concentrations of 527 µg/kg, which is 105.4 times higher than the allowed limit. For animal feed, the limit of 20 µg/kg was exceeded in 40.5% of the samples, with the highest concentration being 26.4 times greater than the maximum allowable level. In 2021, the non-compliance rates for AFB₁ in food and feed were 8.3% and 2.3%, respectively, while in 2023, they were 23.2% and 12.2%, respectively. Fumonisins contamination was also high, particularly in 2021, with fumonisin B₁ (FB₁) detected in 87.1% of samples and average concentrations reaching 4532 µg/kg. Although levels decreased in 2023 (70.7% occurrence, average 885 µg/kg), contamination remained significant. Deoxynivalenol (DON) contamination was consistently high (>70% of samples), with peak concentrations of 606 µg/kg recorded in 2021. Zearalenone (ZEN) and ochratoxin A (OTA) occurred less frequently, but ZEN levels peaked in 2022 at 357.6 µg/kg, which is above the regulatory limit of 350 µg/kg for food. Trichothecenes (HT-2 and T-2 toxins) were detected sporadically, with concentrations well below critical thresholds. Co-occurrence of mycotoxins was frequent, with significant mixtures detected, particularly between aflatoxins and fumonisins, as well as other fusarial toxins. The analysis demonstrated that temperature, humidity, and rainfall during both the growing and harvest seasons strongly influenced mycotoxin levels, with the most severe contamination occurring under specific climatic conditions. Notably, the highest mycotoxin levels, like aflatoxins, were linked to warmer temperatures and lower rainfall. The high non-compliance rates for aflatoxins and fumonisins and co-contamination pose significant food and feed safety risks. From a public health perspective, chronic exposure to contaminated maize increases the likelihood of carcinogenesis and reproductive disorders. Reduced productivity and bioaccumulation in animal tissues/products represent serious economic and safety concerns for livestock. This study provides insights into the potential risks to food and feed safety and the need for enhanced regulatory frameworks, continuous monitoring, and mitigation strategies in Serbia as well as other geographical regions. Full article

Trichothecene type A mycotoxins, such as T-2, HT-2, and diacetoxyscirpenol (DAS), are known to induce cytotoxicity and apoptosis in different cell types. As all three Fusarium toxins may occur concomitantly in a given food or feed commodity, there is growing interest in the effect of such mycotoxin mixtures. This study aimed to identify the toxic interactions among T-2, HT-2, and DAS in a human Jurkat cell model. As a first step, an MTT assay was used to assess cytotoxicity after 24 h of cell exposure to individual mycotoxins and their mixtures. The results were used to calculate the combination index (CI), which indicates the nature of the mycotoxin interactions. In Jurkat T cells, the toxicity ranking for the individual mycotoxins was T-2 > HT-2 > DAS. The CI values of the dual and triple mycotoxin combinations calculated from the results of the MTT and reactive oxygen species assays showed synergistic effects at low concentrations and an apparent antagonism at very high concentrations for all combinations. The additional cytometric analyses confirmed the synergistic effects, as expected, following co-exposure to the three tested trichothecenes. As the lower toxin concentrations investigated reflect natural contamination levels in food and feeds, the synergistic effects identified should be considered in risk characterization for trichothecene exposure in humans and animals. Full article

Background/Objectives: The purpose of this research was to investigate the effects of bisphenol A (BPA) exposure on neurobehavioral testing in young Wistar rats and to evaluate the therapeutic potential of Thymus ciliatus (TEO) essential oil to attenuate the damage induced by this chemical toxin. Methods: The essential oil was extracted by hydro-distillation (yield of 2.26%), and the characterization by GC-MS indicates that the major components of Thymus ciliatus oil are thymol (63.33%), p-cymene (13.4%), and σ-terpinene (6.69%). Acute BPA intoxication was induced with a dose of 50 mg/kg orally for 60 days. The neurobehavioral evaluation, performed using a comprehensive set of tests including the forced swim test, dark/light box, Morris water maze, open field test, and sucrose preference test, clearly demonstrated that bisphenol A (BPA) exposure induced significant neurobehavioral impairments. Results: These impairments included reduced exploratory behavior indicative of heightened stress, anxiety, and depressive-like states, as well as deficits in memory and learning. Furthermore, BPA intoxication was associated with metabolic disturbances such as hyperglycemia along with histopathological evidence of brain tissue damage. However, TEO treatment attenuated these adverse effects by restoring neurobehavioral function. Molecular docking analysis revealed an affinity between the major essential oils identified in T. ciliatus, BPA, and the 5HT2C receptor and the MAO, AChE, and BChE enzymes, suggesting a potential mechanism underlying BPA’s effects on behavior and memory. In addition, TEO also showed an interaction with these molecules, suggesting a therapeutic potential against BPA. These findings underscore the promising role of TEO in mitigating the poisonous effects of BPA and pave the way for additional research into the molecular mechanisms and therapeutic uses of natural bioactive compounds for the prevention and treatment of toxic diseases. Thymol, the major compound in TEO, exhibited activity related to the dopamine and serotonin pathways, so it could have potential antidepressant properties. Conclusions: Thymol might be a promising candidate for the treatment of neurodegenerative and neurological disorders such as depression, Parkinson’s disease, and Alzheimer’s disease while also preventing histological damage in the brain. Full article

Grain industries are interested in an integrated approach to in-silo grain quality and safety management using carbon dioxide (CO₂) measurement with temperature and moisture monitoring. Our study investigates if CO₂ production could predict mycotoxin production (T-2 toxin, HT-2 toxin, its glucoside, and ochratoxin A (OTA)) and identify storage conditions exceeding legislative limits in stored oats for the first time. The influence of water activity (a_w) levels (0.70–0.95 a_w), temperature (15 and 20 °C), and storage duration on (a) Fusarium populations, (b) CO₂ respiration rates (RRs), and (c) mycotoxin concentrations in stored oats was examined. One hundred and twenty samples were analysed for multiple mycotoxins by LC-MS/MS. Substantial differences were found in the RRs of oats at ≥0.90 a_w at both temperatures. A moderate positive correlation between CO₂ and mycotoxins was noticed and mycotoxins exceeded their limits at ≥0.90 a_w (22% moisture content) when RR ≥ 25 µg CO₂ kg⁻¹ h⁻¹. This knowledge forms the basis for developing decision support systems for improving oats’ storage management. Full article

Oats are susceptible to contamination by Fusarium mycotoxins, including deoxynivalenol (DON), zearalenone (ZEN), and T-2/HT-2 toxins, posing food safety risks. This study analyses the variation in levels of 14 mycotoxins in 200 individual oat kernels from two DON-contaminated batch samples (mean = 3498 µg/kg) using LC-MS/MS. The samples also contained deoxynivalenol-3-glucoside (DON-3G), 3-acetyldeoxynivalenol (3-ADON), 15-acetyldeoxynivalenol (15-ADON), and ZEN. Contamination levels varied notably among individual kernels, with DON detected in 70% of them, followed by DON-3G (24.5%) and 3-ADON (20.5%). Importantly, 8% of kernels exceeded the EU legal limit for DON (1750 µg/kg), and some occasionally surpassed limits for ZEN and T-2/HT-2. Correlation analyses revealed strong associations between DON and its derivatives but weaker correlations with other toxins. Mycotoxin ratios varied widely, indicating that although they often co-occur, their concentrations differ between kernels. Contamination did not significantly impact kernel weight, though a slight trend toward lower weights in contaminated kernels was noted. Additionally, sampling statistics showed that as the percentage of selected kernels increased, the probability of batch sample rejection for DON contamination rose significantly. The study highlights the heterogeneity of mycotoxin contamination in oat batches, emphasising the importance of accurate detection and regulatory compliance to ensure safer oat-based products. Full article

The effect of mycotoxin exposure on follicular fluid composition and reproductive outcomes in women undergoing in vitro fertilisation (IVF) was investigated in this study. Twenty-five patients were included, and follicular fluid and serum samples were analysed for various mycotoxins. Principal observations:1. Mycotoxin presence: All examined mycotoxins were detected in follicular fluid. Follicular fluid (ff) levels: Deoxynivalenol (DON), alfa-Zearalenol (α-ZOL), Zearalenone (ZEN), and total aflatoxin (AFs) were significantly higher in follicular fluid than in serum. 2. Follicular fluid and reproductive outcomes: A positive correlation was observed between the ratio of oocytes to total follicles and the follicular Fumonisin B1 (FB1) levels. Multiple linear regression analysis revealed a significant relationship between DON and T-2/HT-2 toxins (T2/HT2) levels in the follicular fluid. 3. Hormone levels: Follicular 17-beta estradiol (E2) and progesterone (P4) levels were higher than the serum levels. Follicular P4 correlated with serum P4 and Anti-Müllerian hormone (AMH) levels. In contrast, follicular E2 did not correlate with plasma E2 levels. 4. Mycotoxin–hormone interactions: A positive correlation was observed between follicular P4 and T2/HT2 toxin levels, whereas a negative correlation was found between ffE2 and ffT2/HT2, and a positive correlation was found between ZEN and E2. Conclusion: This study elucidated the presence of various mycotoxins in the follicular fluid and their potential influence on reproductive outcomes. Further research is warranted to clarify the specific mechanisms underlying these effects and develop strategies for detecting mycotoxin exposure in women undergoing IVF. Full article