Search Results (130)

A blend with pumpkin and sunflower seed flours was prepared and dried at 41.5 °C for 5 h to create a minimally heat-treated blend for a raw food diet. The blend was inoculated with Lactobacillus acidophilus and Fusarium langsethiae to assess the effect of L. acidophilus on Fusarium growth and mycotoxin production. Drying did not affect the content of naturally occurring microorganisms but significantly reduced water activity (p < 0.05) and increased total phenolic content in samples with external microorganisms. Lactobacilli content remained unchanged after drying (4.8 log CFU/g), while F. langsethiae increased by 1.5 log CFU/g. Principal component analysis showed PC1 explained 95.1% of total variance, driven by Fusarium mycotoxin production. A significant difference in total mycotoxin was found between samples with F. langsethiae alone and those with both F. langsethiae and L. acidophilus (p < 0.05). Lactic acid bacteria could reduce fusarium mycotoxin risk in raw food diet mixtures. Full article

The high acidity, alcohol, and mycotoxin levels in distiller’s grains (DGs) limit its application in practical production. To address these issues, a new DG fermentation technique was developed in this research. Firstly, four strains were selected and the fermentation conditions were optimized to ferment the fresh DGs. When the inoculum was set at 8%, the fermentation temperature was maintained at 35 °C, the fermentation time lasted for 48 h, the bacterial mixture ratio (Bacillus subtilis ASAG 216: Lactobacillus acidophilus G1: Saccharomyces cerevisiae ANP 101: Streptococcus thermophilus EFR 046) was 1:1:2:1, and the contents of crude protein in fermented DGs (FDGs) were the highest, so we chose these fermentation conditions to ferment the DGs. In addition, under these fermentation conditions, the amino acids were significantly (p < 0.05) increased while the concentrations of crude fiber and mycotoxins contents were significantly (p < 0.05) decreased in FDGs than in DGs. Subsequently, the nutritional value of DGs and FDGs were evaluated using a two-step in vitro digestion method. The digestibility of dry matter, protein, and crude fiber increased by 16.23%, 13.54%, and 64.09%, respectively, in FDGs compared to that in DGs. Finally, laying hens were treated by adding 0%, 1%, 2%, and 4% FDG to the basal diet for 4 weeks. The results demonstrated that addition of 2% FDG in the diet could significantly (p < 0.05) increase the laying rate of hens compared to that fed the control diet, while addition of 4% FDG in the diet could remarkably (p < 0.05) reduce the rate of broken eggs compared to the other groups. There were no significant (p > 0.05) differences in other indices. These indicates that FDG has potential as a functional feed additive to enhance animal productivity. Full article

Aflatoxins are carcinogenic secondary metabolites produced by Aspergillus species and are common contaminants of many crops including maize. Atoxigenic Aspergillus flavus strains, formulated as biocontrol products such as Aflasafe^® TZ01, that comprises a mixture of four native atoxigenic strains, are used as pre-harvest agents to suppress toxigenic strains and reduce aflatoxin levels. This study assessed the intended and potential unintended impacts of Aflasafe^® TZ01 on mycotoxin contamination in maize. A total of 158 samples 79 from treated and 79 from untreated fields were collected from Chemba and Kiteto districts in Tanzania. Multi-mycotoxin analysis was conducted using liquid chromatography coupled with tandem mass spectrometry (LC-MS/MS). Detected toxins included aflatoxins (AFB1, AFB2, AFG1, AFG2), trichothecenes, and fumonisins (FB1, FB2, FB3). Non-parametric paired t-test analysis showed significant reductions in AFB1 (62%, p = 0.024) in treated samples. The mean concentrations of Fusarium mycotoxins such as NIV, T2, and ZEN were higher in treated maize. However, statistical analysis showed that these differences were only numerical trends, and were not significant (p > 0.05). These findings confirm the efficacy of Aflasafe^® TZ01 in reducing aflatoxins, while underscoring the importance of continued monitoring for other mycotoxins as part of integrated mycotoxin management strategies to mitigate both aflatoxins and co-occurring toxins. Full article

Mycotoxins are toxic secondary metabolites produced by various fungi that contaminate livestock feed, posing serious threats to animal health, productivity, and food safety. Although historical research has often examined individual mycotoxins in isolation, real-world conditions typically involve the simultaneous presence of multiple mycotoxins, resulting in additive or synergistic toxic effects that are often more severe than those observed with single toxin exposures. This review comprehensively synthesizes recent findings on multi-mycotoxin contamination in livestock feed, highlighting their physiological effects, mechanisms of action, and implications for regulatory frameworks. Multi-mycotoxin interactions exacerbate oxidative stress, immune suppression, impaired reproduction, and organ damage across species, leading to reduced growth performance, decreased milk and egg production, compromised carcass and wool quality, and increased mortality rates. A major concern is that current international regulatory standards mainly address individual mycotoxins, overlooking the compounded risks of co-occurrence. Global surveillance studies consistently reveal high prevalence rates of mycotoxin mixtures in feedstuffs, especially combinations involving DON, ZEN, AFB₁, FB₁, and OTA. Understanding these interactions and their underlying cellular mechanisms is critical for improving risk assessment models, formulating integrated mitigation strategies, and safeguarding both livestock productivity and human food security. Full article

In this study, the chemopreventive effects of rosmarinic acid (RA), a major phenolic acid of the plant Rosmarinus officinalis L., against the carcinogenic naturally occurring mycotoxin aflatoxin B1 (AFB1) were investigated using both in silico and in vitro approaches. The in silico investigation of the chemical reactions between rosmarinic acid and the carcinogenic metabolite of AFB1, aflatoxin B1 exo-8,9-epoxide (AFBO), was conducted by activation free energies calculations with DFT functionals M11-L and MN12-L, in conjunction with the 6-311++G(d,p) flexible basis set and implicit solvation model density (SMD), according to a newly developed quantum mechanics-based protocol for the evaluation of carcinogen scavenging activity (QM-CSA). Following the computational analyses, the chemoprotective effects of RA were further studied in vitro in human hepatocellular carcinoma HepG2 cells by analyzing its influence on AFB1-induced genotoxicity using a comet assay, γH2AX, and p-H3, while its impact on cell proliferation and cell cycle modulation was assessed using flow cytometry. Our computational results revealed that the activation free energy required for the reaction of RA with AFBO (14.86 kcal/mol) is significantly lower than the activation free energy for the competing reaction of AFBO with guanine (16.88 kcal/mol), which indicates that RA acts as an efficient natural scavenger of AFBO, potentially preventing AFB1-specific DNA adduct formation. The chemoprotective activity of RA was confirmed through in vitro experiments, which demonstrated a statistically significant (p < 0.05) reduction in AFB1-induced single- and double-strand breaks in HepG2 cells exposed to a mixture of AFB1 and RA at non-cytotoxic concentrations. In addition, RA reversed the AFB1-induced reduction in cell proliferation. Full article

The replacement of ingredients from animal sources with plant-based ingredients is increasing the risk of contamination by mycotoxins in aquafeeds, potentially causing detrimental effects on fish welfare. However, limited research has been carried out so far on the impact of mycotoxins on fish health. Hence, the aim of this study was to assess the toxicological effects of the dietary emerging (enniatin B, ENNB) and regulated (fumonisin B1, FB1) mycotoxins (150 µg/kg) in different tissues of juvenile gilthead seabream (Sparus aurata) after 28 days of dietary exposure. Fitness indexes, plasma metabolites, and biomarkers of oxidative stress, metabolism, cellular, and neurotoxic damage were assessed. The exposure to each mycotoxin was sufficient to cause distinct effects in fish tissues. ENNB appears to be the most harmful mycotoxin to S. aurata, inducing changes on alkaline phosphatase and lipase activities in plasma, as well as protein and lipid degradation in liver. Increased lipid degradation was also induced in the brain by FB1 alone or combined with ENNB, whereas the exposure to the mixture inhibited acetylcholinesterase activity. Overall, this study contributes by highlighting the toxicological attributes of ENNB, thus reinforcing the need to include this mycotoxin in future legislation. Full article

Mycotoxins are toxic secondary metabolites produced by fungi that often contaminate food materials used in beer production, posing health risks to consumers. This study investigated the occurrence and levels of mycotoxins in commercial beers commercially available in São Paulo, Brazil, and assessed the estimated daily intake (EDI) of quantifiable mycotoxins. Sixty beer samples from different brands and compositions (barley malt, malt with corn, and malt with rice) were analyzed for deoxynivalenol (DON); aflatoxins (AFs) B₁, B₂, G₁, and G₂; ochratoxin A (OTA); T-2 toxin; fumonisins (F) (B₁ and B₂); and zearalenone (ZEN) using ultra-performance liquid chromatography coupled to tandem mass spectrometry. FB₁ was quantified in all samples, while DON, ZEN, OTA, AFB₁, and T-2 toxin were detected in 40, 65, 25, 20, and 10%, respectively. Mean levels of 2.38, 36.41, 0.19, 1.05, 0.78, and 0.47 ng/mL were observed for FB₁, DON, ZEN, OTA, AFB₁, and T-2 toxin, respectively. Mycotoxin co-occurrence was observed in 43 (71.7%) samples analyzed, with DON and FB₁ as the most frequent combination (20%). The EDI values of individual mycotoxins were generally below tolerable daily intakes established by international agencies. However, the co-occurrence of up to four different mycotoxins in beers warrants concern on the possible interactive toxic effects of mycotoxin mixtures and reinforces the necessity of specific regulations for ready-to-drink beverages in Brazil. Full article

A total of 98 samples of spices, herbs, and mixtures commercialized in Italy were analyzed for 11 mycotoxins, regulated and non-regulated. The occurrence of 1–4 mycotoxins was found in 84% and 60% of spice samples and herb samples, respectively. Spices were the most contaminated matrix. Total aflatoxins were detected in 5% and 9% of herbs and spices, respectively, and ochratoxin A was detected in 14% of spices and not at all in herbs. Only one sample of spices (cloves) showed an AFB₁ value (9.0 µg/kg) higher than the limit. No sample of herbs and spices had an OTA content beyond the limit. Within the non-regulated mycotoxins, ZEA was the most commonly occurring, and its mean levels in positive samples of spices ranged from 4.6 µg/kg (ZEA) to 1813.9 µg/kg (FB₁). The mean levels of FB₂ and ZEA in positive samples of herbs were 131.4 µg/kg and 2.5 µg/kg, respectively. The mean levels of non-regulated mycotoxins in positive samples of herbs-and-spices mixtures ranged from 2.6 µg/kg (ZEA) to 1071.7 µg/kg (FB₁). The most contaminated herbs and spices were garlic and basil, respectively. This study provides a significant amount of information on the natural occurrence of multi-mycotoxins in herbs and spices consumed in Italy, which will be useful for the future regulation of some mycotoxins, in particular, FBs. 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

This study assessed the cytotoxicity of the individual and combined exposure to the fungicide azoxystrobin (AZX) and the three common mycotoxins found in food: ochratoxin A (OTA), deoxynivalenol (DON), and T-2 toxin. Cytotoxic effects were evaluated using the resazurin and MTT assays in human hepatocarcinoma (HepG2) cells after 24 h of exposure, and the type of interaction between the compounds was determined using the isobologram method. Results showed that T-2 was the most cytotoxic compound, followed by DON, OTA, and AZX. The compound ratios in the mixture were calculated using three sublethal concentrations (IC_50/2, IC_50/4, and IC_50/8) to achieve equal toxicity for each compound. Interaction analysis revealed that the nature of the interaction varied across components and concentrations. The AZX and DON mixture produced an antagonistic effect at all the analyzed effect levels. AZX and OTA or T2 mixtures, and tertiary combinations displayed antagonism at low effect values but additivity at high effect levels. Importantly, the quaternary mixture demonstrated synergism at all the effect levels. These findings highlight that the co-occurrence of fungicides and mycotoxins in food commodities can lead to complex exposure scenarios that may result in combined toxic effects on the organism. Full article

Mycotoxins are toxic compounds found in food and feed that pose significant risks to human and animal health. This work reviews recent studies on the cytotoxic effects of four mycotoxins: beauvericin (BEA), citrinin (CTN), moniliformin (MON), and patulin (PAT) in various cell lines. Additionally, an experimental study evaluates the effects of these mycotoxins and their binary combinations on human neuroblastoma cells (SH-SY5Y) after 24 and 48 h of exposure using the 3-(4,5-Dimethylthiazol-2-yl)-2,5-Diphenyltetrazolium Bromide (MTT) assay. This analysis is driven by the additional risks posed by the frequent occurrence of these combinations in agricultural and food products, as well as the lack of studies addressing their effects, interactions, and regulatory frameworks. This research focuses on comparing the cytotoxicity data obtained in the SH-SY5Y cell line with previously reported findings in the literature for other cell lines exposed to BEA, CTN, MON, and PAT, individually and in binary combination. The literature highlights significant scientific interest in understanding the cytotoxic effects of these mycotoxins, with findings varying based on exposure time and concentration. Experimentally, PAT demonstrated the highest toxicity in SH-SY5Y cells, while MON was the least toxic. Among combinations, BEA + MON and CTN + PAT showed the greatest reduction in cell viability. However, medium inhibitory concentration (IC₅₀) values were not reached for most combinations involving MON, reflecting its lower potency under the studied conditions. These findings underscore the importance of further investigation and enhanced regulations to address the health risks posed by mycotoxins, as their cytotoxic effects remain a pressing issue in food safety. Full article

This study investigated the impact of baking conditions on mycotoxin content in Greek pita bread, a single-layered flatbread baked at high temperatures for short time intervals. Dough samples were artificially contaminated with a multi-mycotoxin mixture, including aflatoxins (AFs) G2, G1, B2, and B1; ochratoxin A (OTA); deoxynivalenol (DON); and zearalenone (ZEA). Flatbreads were baked under three temperature–time combinations (220, 270, and 320 °C for 4, 2, and 1 min, respectively), with additional evaluation of a parbaking process (baking halted at 75% of the total time for each respective temperature, bread was stored at −18 °C, then bread was baked for 3 min at 180 °C). A QuEChERS-LC-MS/MS method was implemented for the determination of mycotoxins. The results demonstrated varying degrees of thermal degradation, with AFs B1 and G1 showing the highest decrease (39% on average), followed by AFG2, AFB2, ZEA, and DON (16–25%), while OTA remained relatively thermostable. Multivariate analyses classified flatbreads into two groups: higher baking temperatures and parbaking favored reductions in AFG1, AFG2, ZEA, OTA, and AFB2 levels while longer baking times at lower temperatures favored DON and AFB1 reduction. These findings provide insights for optimizing baking conditions to improve food safety in industrial and home-baking applications. Full article

Plant-based materials are increasingly being used as ingredients of aquaculture feed. These materials are prone to mycotoxin contamination, as mycotoxigenic fungi infest crop plants in the field and agricultural products during storage. As mycotoxins can cause toxic effects in aquatic animals, their occurrence in feedstuffs should be monitored. To this end, we performed an extensive global survey of mycotoxin contamination in aquaculture feed and plant-based feed raw materials. We collected samples of compound feed for fish (n = 226) and shrimps (n = 61), maize (n = 3448), maize DDGS (n = 149), wheat (n = 1578), soybean (n = 428), and rice (n = 65). We analyzed concentrations of 51 mycotoxins, emerging mycotoxins, masked mycotoxins, and mycotoxin metabolites. Mycotoxins were almost ubiquitously present in compound feed, as >90% of samples were contaminated with at least one mycotoxin. Feed raw materials exhibited distinct mycotoxin occurrence patterns consistent with known susceptibility to fungal pathogens and with their production process. Unsafe concentrations of aflatoxin B₁ exceeding the EU maximum level were detected in 7.2% of fish feed samples. While most feedstuffs complied with EU guidance values for deoxynivalenol, zearalenone, and fumonisins, a comparison of detected concentrations with dietary concentrations reported to cause adverse effects in fish and shrimps in published studies indicated that significant fractions of samples contained potentially harmful levels of these mycotoxins. In addition to regulated mycotoxins, several emerging mycotoxins (e.g., enniatins, beauvericin, alternariol, moniliformin) were prevalent. Feed was frequently co-contaminated with multiple mycotoxins indicating a risk of combined effects. In conclusion, mycotoxin contamination was common in aquaculture feed and fractions of samples were contaminated with mycotoxin levels known to exert adverse effects in aquaculture species. Results of this survey highlight the necessity for targeted studies on the effects of frequently detected mycotoxin mixtures and emerging mycotoxins in fish and shrimp. Full article