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11 pages, 486 KiB  
Article
Multi-Mycotoxin Contamination of Concentrates Fed to Dairy Calves in Southeast Brazil: A Case Report
by Rogério D’Antonio Pires, Aline Moreira Borowsky, Tobias Alves e Silva, Giovanna Canela Ruiz Castro Evangelista, Carla Maris Machado Bittar and Carlos Humberto Corassin
Dairy 2025, 6(4), 44; https://doi.org/10.3390/dairy6040044 - 4 Aug 2025
Viewed by 262
Abstract
The diet of dairy calves can be contaminated with mycotoxins, posing a potential risk to animal health. This case study report aimed to make the first assessment of the presence of multiple mycotoxins in concentrates fed to dairy calves in Brazil. A total [...] Read more.
The diet of dairy calves can be contaminated with mycotoxins, posing a potential risk to animal health. This case study report aimed to make the first assessment of the presence of multiple mycotoxins in concentrates fed to dairy calves in Brazil. A total of 19 concentrate samples intended for dairy calves were analyzed using liquid chromatography coupled with mass spectrometry. Aflatoxins, deoxynivalenol, and T-2 toxin were not detected in any samples, whereas fumonisins B1 (FB1) and B2 (FB2) were present in 100% of the samples, with mean concentrations of 2750.1 μg/kg and 834.9 μg/kg, respectively. Zearalenone (ZEN) was detected in 36.8% of samples, with a mean concentration of 929.9 μg/kg. Significant correlations were observed between FB1 and FB2 (ρ = 0.978; p < 0.001) and between FB2 and ZEN (ρ = 0.735; p = 0.05). While the physical form of the concentrate did not influence (p > 0.05) mycotoxin concentrations, a trend was observed for FB1 (ρ = −0.417; p = 0.07) and FB2 (ρ = −0.395; p = 0.09). These findings highlight the frequent occurrence of Fusarium mycotoxins, likely due to pre-harvest contamination, emphasizing the potential risk of additive or synergistic effects in dairy calves. Full article
(This article belongs to the Section Dairy Animal Nutrition and Welfare)
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14 pages, 4802 KiB  
Article
Curcumin Attenuates Zearalenone-Induced Reproductive Damage in Mice by Modulating the Gut Microbe–Testis Axis
by Bangwang Peng, Shuaiju Guo, Junlong Niu, Yongpeng Guo, Zhixiang Wang and Wei Zhang
Foods 2025, 14(15), 2703; https://doi.org/10.3390/foods14152703 - 31 Jul 2025
Viewed by 268
Abstract
Zearalenone (ZEN), a mycotoxin commonly found in cereal crops and foods, induces testicular damage and disrupts gut microbial composition. Curcumin (CUR), a bioactive compound derived from turmeric, is known to enhance intestinal microbial balance and exhibit anti-inflammatory properties. This study aimed to investigate [...] Read more.
Zearalenone (ZEN), a mycotoxin commonly found in cereal crops and foods, induces testicular damage and disrupts gut microbial composition. Curcumin (CUR), a bioactive compound derived from turmeric, is known to enhance intestinal microbial balance and exhibit anti-inflammatory properties. This study aimed to investigate the mechanism by which CUR alleviates ZEN-induced reductions in sperm quality through the modulation of the gut microbiota–testis axis. Forty-eight 6-week-old Balb/c male mice were randomly assigned to four treatment groups: control (CON), CUR (200 mg/kg body weight CUR), ZEN (40 mg/kg body weight ZEN), and ZEN + CUR (200 mg/kg CUR + 40 mg/kg ZEN). The degree of sperm damage was quantified by assessing both the survival rate and the morphological integrity of the spermatozoa. CUR was found to mitigate ZEN-induced reductions in the testosterone levels, testicular structural damage, and disrupted spermatogenesis. Exposure to ZEN markedly perturbed the gut microbiota, characterized by increased relative abundances of Prevotella and Bacteroides and a concomitant reduction in Lactobacillus. These alterations were accompanied by pronounced activation of the IL-17A–TNF-α signaling axis, as demonstrated by elevated transcriptional and translational expression of pathway-associated genes and proteins. Co-administration of CUR effectively reinstated microbial homeostasis and mitigated ZEN-induced IL-17A pathway activation. In conclusion, ZEN induces testicular inflammation and reduced sperm quality by lowering testosterone levels and disrupting gut microbial balance, which drives the testicular IL-17A signaling pathway. CUR alleviates ZEN-induced testicular inflammation and sperm quality reduction by restoring beneficial gut microbes and testosterone levels. Full article
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18 pages, 3415 KiB  
Article
Study on the Modification of Dietary Fiber and Degradation of Zearalenone in Corn Germ Meal by Solid-State Fermentation with Bacillus subtilis K6
by Jiahao Li, Kailong Li, Langwen Tang, Chun Hua, Na Chen, Chenxian Yang, Ying Xin and Fusheng Chen
Foods 2025, 14(15), 2680; https://doi.org/10.3390/foods14152680 - 30 Jul 2025
Viewed by 300
Abstract
Although corn germ meal is a rich source of dietary fiber, it contains a relatively low proportion of soluble dietary fiber (SDF) and is frequently contaminated with high levels of zearalenone (ZEN). Solid-state fermentation has the dual effects of modifying dietary fiber (DF) [...] Read more.
Although corn germ meal is a rich source of dietary fiber, it contains a relatively low proportion of soluble dietary fiber (SDF) and is frequently contaminated with high levels of zearalenone (ZEN). Solid-state fermentation has the dual effects of modifying dietary fiber (DF) and degrading mycotoxins. This study optimized the solid-state fermentation process of corn germ meal using Bacillus subtilis K6 through response surface methodology (RSM) to enhance SDF yield while efficiently degrading ZEN. Results indicated that fermentation solid-to-liquid ratio and time had greater impacts on SDF yield and ZEN degradation rate than fermentation temperature. The optimal conditions were determined as temperature 36.5 °C, time 65 h, and solid-to-liquid ratio 1:0.82 (w/v). Under these conditions, the ZEN degradation rate reached 96.27 ± 0.53%, while the SDF yield increased from 9.47 ± 0.68% to 20.11 ± 1.87% (optimizing the SDF/DF ratio from 1:7 to 1:3). Scanning electron microscopy (SEM) and confocal laser scanning microscope (CLSM) revealed the structural transformation of dietary fiber from smooth to loose and porous forms. This structural modification resulted in a significant improvement in the physicochemical properties of dietary fiber, with water-holding capacity (WHC), oil-holding capacity (OHC), and water-swelling capacity (WSC) increasing by 34.8%, 16.4%, and 15.2%, respectively. Additionally, the protein and total phenolic contents increased by 23.0% and 82.61%, respectively. This research has achieved efficient detoxification and dietary fiber modification of corn germ meal, significantly enhancing the resource utilization rate of corn by-products and providing technical and theoretical support for industrial production applications. Full article
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22 pages, 1271 KiB  
Article
Toxigenic Fungi and Co-Occurring Mycotoxins in Maize (Zea mayz L.) Samples from the Highlands and Coast of Ecuador
by Héctor Palacios-Cabrera, Juliana Fracari, Marina Venturini Copetti, Carlos Augusto Mallmann, Marcelo Almeida, María Raquel Meléndez-Jácome and Wilson Vásquez-Castillo
Foods 2025, 14(15), 2630; https://doi.org/10.3390/foods14152630 - 26 Jul 2025
Viewed by 404
Abstract
Maize is a key crop in Ecuador for both human and animal consumption. Its vulnerability to fungal contamination and mycotoxins poses risks to food safety. The aim of this study was to analyze the occurrence of fungi and mycotoxins in maize grown in [...] Read more.
Maize is a key crop in Ecuador for both human and animal consumption. Its vulnerability to fungal contamination and mycotoxins poses risks to food safety. The aim of this study was to analyze the occurrence of fungi and mycotoxins in maize grown in different regions of Ecuador (29 localities) and postharvest factors influencing contamination. Fungal identification was performed through culturing and morphological analysis. Analysis of multi-toxins was carried out using liquid chromatography coupled with mass spectrometry (LC-MS/MS). Statistical analyses included PCA and linear regression models. Fungal contamination was found in 93.3% of samples; mycotoxins were present in 90%. Fusarium and Aspergillus were dominant. Fumonisins (66.6%), zearalenone (30%), aflatoxins (16.7%), and trichothecenes B (13.3%) were the most prevalent. Co-occurrence of up to three mycotoxins per sample was observed, more frequent on the coast. Grain moisture and temperature were strongly correlated with contamination levels. The study reveals widespread contamination of Ecuadorian maize, with environmental and postharvest factors playing key roles. This poses a food safety concern, highlighting the need for improved storage and monitoring systems. Full article
(This article belongs to the Special Issue Mycotoxins in Foods: Occurrence, Detection, and Control)
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20 pages, 1243 KiB  
Article
Comparison of Capillary Electrophoresis and HPLC-Based Methods in the Monitoring of Moniliformin in Maize
by Sara Astolfi, Francesca Buiarelli, Francesca Debegnach, Barbara De Santis, Patrizia Di Filippo, Donatella Pomata, Carmela Riccardi and Giulia Simonetti
Foods 2025, 14(15), 2623; https://doi.org/10.3390/foods14152623 - 26 Jul 2025
Viewed by 188
Abstract
Over the past few decades, scientific interest in mycotoxins—fungal metabolites that pose serious concern to food safety, crop health, and both human and animal health—has increased. While major mycotoxins such as aflatoxins, ochratoxins, deoxynivalenol, fumonisins, zearalenone, citrinin, patulin, and ergot alkaloids are well [...] Read more.
Over the past few decades, scientific interest in mycotoxins—fungal metabolites that pose serious concern to food safety, crop health, and both human and animal health—has increased. While major mycotoxins such as aflatoxins, ochratoxins, deoxynivalenol, fumonisins, zearalenone, citrinin, patulin, and ergot alkaloids are well studied, emerging mycotoxins remain underexplored and insufficiently investigated. Among these, moniliformin (MON) is frequently detected in maize-based food and feed; however, the absence of regulatory limits and standardized detection methods limits effective monitoring and comprehensive risk assessment. The European Food Safety Authority highlights insufficient occurrence and toxicological data as challenges to regulatory development. This study compares three analytical methods—CE-DAD, HPLC-DAD, and HPLC-MS/MS—for moniliformin detection and quantification in maize, evaluating linear range, correlation coefficients, detection and quantification limits, accuracy, and precision. Results show that CE-DAD and HPLC-MS/MS provide reliable and comparable sensitivity and selectivity, while HPLC-DAD is less sensitive. Application to real samples enabled deterministic dietary exposure estimation based on consumption, supporting preliminary risk characterization. This research provides a critical comparison that supports the advancement of improved monitoring and risk assessment frameworks, representing a key step toward innovating the detection of under-monitored mycotoxins and laying the groundwork for future regulatory and preventive measures targeting MON. Full article
(This article belongs to the Special Issue Recent Advances in the Detection of Food Contaminants and Pollutants)
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22 pages, 1009 KiB  
Review
Mycotoxin-Caused Intestinal Toxicity: Underlying Molecular Mechanisms and Further Directions
by Tian Li, Weidong Qiao, Jiehong Zhou, Zhihui Hao, Gea Oliveri Conti, Tony Velkov, Shusheng Tang, Jianzhong Shen and Chongshan Dai
Toxics 2025, 13(8), 625; https://doi.org/10.3390/toxics13080625 - 26 Jul 2025
Viewed by 457
Abstract
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 [...] Read more.
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
(This article belongs to the Topic Recent Advances in Veterinary Pharmacology and Toxicology)
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19 pages, 2696 KiB  
Article
Cell Type-Specific Effects of Fusarium Mycotoxins on Primary Neurons and Astroglial Cells
by Viktória Szentgyörgyi, Brigitta Tagscherer-Micska, Anikó Rátkai, Katalin Schlett, Norbert Bencsik and Krisztián Tárnok
Toxins 2025, 17(8), 368; https://doi.org/10.3390/toxins17080368 - 25 Jul 2025
Viewed by 316
Abstract
Fumonisin B1, deoxynivalenol (DON), and zearalenone (ZEA) are toxic secondary metabolites produced by Fusarium molds. These mycotoxins are common food and feed pollutants and represent a risk to human and animal health. Although the mycotoxins produced by this genus can cross the blood–brain [...] Read more.
Fumonisin B1, deoxynivalenol (DON), and zearalenone (ZEA) are toxic secondary metabolites produced by Fusarium molds. These mycotoxins are common food and feed pollutants and represent a risk to human and animal health. Although the mycotoxins produced by this genus can cross the blood–brain barrier in many species, their effect on neuronal function remains unclear. We investigated the cell viability effects of these toxins on specified neural cell types, including mouse primary neuronal, astroglial, and mixed-cell cultures 24 or 48 h after mycotoxin administration. DON decreased cell viability in a dose-dependent manner, independent of the culture type. Fumonisin B1 was toxic in pure neuronal cultures only at high doses, but toxicity was attenuated in mixed and pure astroglial cultures. ZEA had significant effects on all culture types in 10 nM by increasing cell viability and network activity, as revealed by multi-electrode array measurements. Since ZEA is a mycoestrogen, we analyzed the effects of ZEA on the expression of estrogen receptor isotypes ERα and ERβ and the mitochondrial voltage-dependent anion channel via qRT-PCR. In neuronal and mixed cultures, ZEA administration decreased ERα expression, while in astroglial cultures, it induced the opposite effect. Thus, our results emphasize that Fusarium mycotoxins act in a cell-specific manner. Full article
(This article belongs to the Section Mycotoxins)
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20 pages, 2494 KiB  
Article
Effect of Environmental Exposure to Zearalenone on the Metabolic Profile of Patients with Sigmoid Colorectal Cancer or Colorectal Cancer on the Day of Hospital Admission
by Sylwia Lisieska-Żołnierczyk, Magdalena Gajęcka, Łukasz Zielonka, Katarzyna E. Przybyłowicz and Maciej T. Gajęcki
Int. J. Mol. Sci. 2025, 26(14), 6967; https://doi.org/10.3390/ijms26146967 - 20 Jul 2025
Viewed by 332
Abstract
Colorectal cancer is one of the most commonly diagnosed types of cancer and constitutes the second most frequent cancer in women (W) and the third most frequent cancer in men (M). The aim of the study was to determine if environmental exposure to [...] Read more.
Colorectal cancer is one of the most commonly diagnosed types of cancer and constitutes the second most frequent cancer in women (W) and the third most frequent cancer in men (M). The aim of the study was to determine if environmental exposure to zearalenone (ZEN) (a mycoestrogen) affects the metabolic profile of patients diagnosed with sigmoid colorectal cancer (SCC) and colorectal cancer (CRC) (division based on their location) at hospital admission. Male and female patients who were diagnosed with SCC or CRC and whose blood samples tested positive or negative for ZEN participated in a year-long study. Seventeen patients with symptoms of SCC and CRC, in whom ZEN and its metabolites were not detected in peripheral blood, constituted the patients without ZEN (PWZ) group. The experimental groups comprised a total of 16 patients who were diagnosed with SCC or CRC and tested positive for ZEN but negative for ZEN metabolites. Patients exposed to ZEN were characterized by increased levels of liver enzymes (alanine aminotransferase (ALT) from 5.8 to 18.1 IU/L; aspartate aminotransferase (AST) from 2.8 to 10.7 IU/L) and decrease in the value of the De Ritis ratio (below 1.0), different gamma glutamyl transpeptidase and AST activity, lower albumin (from 0.24 g/dL in M to 0.67 g/dL in W) and total protein levels (from 0.75 to 1.76 g/dL), a decrease in total cholesterol (from 21.6 to 40.3 mg/dL) and triglyceride levels (from 7.8 to 37.2 mg/dL), and lower activity of lipase C (from 28.72 to 64.75 IU/L). The metabolic profile of M and W patients diagnosed with SCC and CRC and exposed to ZEN revealed intensified biotransformation processes in the liver, liver damage, and a predominance of catabolic processes. Full article
(This article belongs to the Section Molecular Pathology, Diagnostics, and Therapeutics)
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17 pages, 1217 KiB  
Article
mRNA Expression of Two Colon Enzymes in Pre-Pubertal Gilts During a 42-Day Exposure to Zearalenone
by Magdalena Gajęcka, Łukasz Zielonka and Maciej T. Gajęcki
Toxins 2025, 17(7), 357; https://doi.org/10.3390/toxins17070357 - 17 Jul 2025
Viewed by 324
Abstract
The aim of this study was to determine whether a low dose of zearalenone (ZEN) affects the mRNA expression of the CYP1A1 (P450 cytochrome) and GSTπ1 (glutathione S-transferase) genes in the large intestine of pre-pubertal gilts. Materials: Control (C) group gilts (n [...] Read more.
The aim of this study was to determine whether a low dose of zearalenone (ZEN) affects the mRNA expression of the CYP1A1 (P450 cytochrome) and GSTπ1 (glutathione S-transferase) genes in the large intestine of pre-pubertal gilts. Materials: Control (C) group gilts (n = 18) received a placebo. Experimental (E) group gilts (n = 18) were orally administered 40 μg ZEN/kg body weight (BW) each day before morning feeding for 42 days. Three animals from each group were sacrificed each week of the study. Tissue samples were collected from the medial parts of the ascending colon and the descending colon on six dates. Results: Zearalenone concentrations were multiple times higher in the last three weeks of exposure, and ZEN metabolites were not detected. In phase I, CYP1A1 mRNA expression in the ascending colon was suppressed in the final three weeks of exposure, which substantially increased the ZEN concentration in the descending colon. In phase II, ZEN levels were high in the descending colon due to CYP1A1 suppression in the ascending colon. Consequently, the phase II detoxification processes could not take place due to the absence of a substrate. Conclusion: This study demonstrated that low-dose ZEN mycotoxicosis disrupts the expression of the CYP1A1 and GSTπ1 genes, which co-participate in the enzymatic biotransformation of ZEN in both examined sections of the large intestine. The above could have contributed to increased ZEN accumulation in the mucosa of the descending colon in the last three weeks of exposure. Full article
(This article belongs to the Section Mycotoxins)
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18 pages, 5095 KiB  
Article
Fusarium Species Infecting Greenhouse-Grown Cannabis (Cannabis sativa) Plants Show Potential for Mycotoxin Production in Inoculated Inflorescences and from Natural Inoculum Sources
by Zamir K. Punja, Sheryl A. Tittlemier and Sean Walkowiak
J. Fungi 2025, 11(7), 528; https://doi.org/10.3390/jof11070528 - 16 Jul 2025
Viewed by 929
Abstract
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. [...] Read more.
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
(This article belongs to the Special Issue Plant Pathogens and Mycotoxins)
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24 pages, 3430 KiB  
Article
Differential Impact of Zearalenone on Hepatic Glucose and Lipid Metabolism in Healthy and Ketotic Dairy Cows: An In Vitro Study
by Justyna Barć, Zygmunt Maciej Kowalski and Wojciech Jagusiak
Int. J. Mol. Sci. 2025, 26(14), 6827; https://doi.org/10.3390/ijms26146827 - 16 Jul 2025
Viewed by 177
Abstract
Zearalenone (ZEN), a mycotoxin commonly present in maize-based feed, poses a health risk to dairy cows. While the hepatic effects of ZEN are increasingly studied, little is known about its impact on cows with altered energy metabolism. This study investigated the transcriptional response [...] Read more.
Zearalenone (ZEN), a mycotoxin commonly present in maize-based feed, poses a health risk to dairy cows. While the hepatic effects of ZEN are increasingly studied, little is known about its impact on cows with altered energy metabolism. This study investigated the transcriptional response of liver cells isolated from healthy and ketotic cows to ZEN exposure using a novel in vitro model. Hepatocytes were obtained via biopsy from 12 cows, cultured under standardized conditions, and exposed to ZEN (0–100 µM) for 1, 3, and 6 h. Gene expression analysis focused on targets related to glucose and lipid metabolism. ZEN induced time- and dose-dependent changes in gene expression, with the most prominent effects observed after 1 h. Key metabolic genes were differentially regulated depending on the cow’s metabolic status. Notably, hepatocytes from healthy cows showed a stronger transcriptional response than those from ketotic cows, indicating reduced metabolic adaptability in energy-compromised animals. Significant interaction effects between ZEN dose and metabolic status were observed, especially for genes involved in glycolysis and fatty acid metabolism. This study presents a novel in vitro model and emphasizes the need to consider metabolic health when assessing the risks of mycotoxin exposure in dairy cattle. Full article
(This article belongs to the Section Molecular Toxicology)
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23 pages, 2930 KiB  
Article
Assessment of Nontoxic Surfactant-Modified Kaolinite for Potential Application as an Adsorbent for Mycotoxins
by Milica Ožegović, Marija Marković, Aleksandra Daković, Milena Obradović, Danijela Smiljanić, George E. Rottinghaus, Vesna Jaćević, Ljubiša Ignjatović and Ivana Sredović Ignjatović
Minerals 2025, 15(7), 731; https://doi.org/10.3390/min15070731 - 12 Jul 2025
Viewed by 335
Abstract
In this study, natural kaolin was modified with hexadecyltrimethylammonium bromide (HDTMA-Br) at two levels corresponding to 50% and 90% of its cation exchange capacity. The resulting materials, designated as HKR-50 and HKR-90, were used as adsorbents for the mycotoxins ochratoxin A (OCHRA) and [...] Read more.
In this study, natural kaolin was modified with hexadecyltrimethylammonium bromide (HDTMA-Br) at two levels corresponding to 50% and 90% of its cation exchange capacity. The resulting materials, designated as HKR-50 and HKR-90, were used as adsorbents for the mycotoxins ochratoxin A (OCHRA) and zearalenone (ZEN). The characterization of the HKRs with several methods (X-ray diffraction, DRIFT spectroscopy, thermal analysis (DTA/TG), SEM, zeta potential measurements, and the determination of the point of zero charge and textural properties) confirmed the presence of surfactant ions on the organokaolinites’ surfaces. The adsorption of ZEN and OCHRA by HKRs followed nonlinear adsorption isotherms, suggesting a complex adsorption mechanism. The adsorption capacities of ZEN and OCHRA were similar for HKR-50 and HKR-90 at pH 3, with higher adsorption observed for ZEN (~13.0 mg/g for HKR-50 and HKR-90 for ZEN and ~8.0 mg/g for HKR-50 and HKR-90 for OCHRA). At pH 7, the adsorption of ZEN and OCHRA was lower than at pH 3, especially for OCHRA, but slightly increased with increased amounts of surfactant on the kaolinite surface (8.5 mg/g for HKR-50 and 10.8 mg/g for HKR-90 for ZEN and 2.6 mg/g for HKR-50 and 4.1 mg/g for HKR-90 for OCHRA). Special attention was paid to the safety assessment of the natural kaolin and HKR-90, and toxicological tests confirmed the safety of both materials, as no adverse effects were observed in rats. Full article
(This article belongs to the Special Issue Organo-Clays: Preparation, Characterization and Applications)
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15 pages, 1732 KiB  
Article
Differentiating Zeranol Implant Abuse and Fusarium spp. Toxin-Contaminated Corn Intake by Detection and Quantification of Resorcylic Acid Lactones in Bovine Urine
by Rafael Silva Gomes, Vanessa Gonçalves dos Santos, Carlos Juliano da Silva, Amanda Martinez Nagato Simões, Eliene Alves dos Santos, Mary Ane Gonçalves Lana, Kelly Moura Keller, Marco Blokland, Ane Arrizabalaga-Larrañaga, Rafael Romero Nicolino, Marcelo Resende de Souza, Tadeu Chaves de Figueiredo, Saskia Sterk and Silvana de Vasconcelos Cançado
Toxins 2025, 17(7), 347; https://doi.org/10.3390/toxins17070347 - 11 Jul 2025
Viewed by 417
Abstract
Resorcylic acid lactones (RALs) are fungal metabolites with known biological activity. Zeranol, a synthetic RAL, has been used as an estrogenic growth promoter in cattle; however, its use is prohibited in several countries. Zearalenone, a mycotoxin produced by Fusarium spp., is commonly found [...] Read more.
Resorcylic acid lactones (RALs) are fungal metabolites with known biological activity. Zeranol, a synthetic RAL, has been used as an estrogenic growth promoter in cattle; however, its use is prohibited in several countries. Zearalenone, a mycotoxin produced by Fusarium spp., is commonly found in contaminated animal feed and can be metabolized into other RALs, which are subsequently excreted in urine. To differentiate between natural contamination from feed and the illegal administration of zeranol, the European Union Reference Laboratory for Growth Promoters (EURL) developed a mathematical equation. This study aims to evaluate the detection and quantification of RALs in bovine urine from animals fed zearalenone-contaminated diets, implanted with zeranol, or subjected to both conditions. RALs were detected and quantified in the urine of cattle consuming contaminated corn, while zeranol and taleranol were identified in the urine of implanted animals. The EURL equation proved to be a valuable tool for determining the origin of RALs in bovine urine and holds significant potential for monitoring and enforcing regulations regarding the illegal use of zeranol. Full article
(This article belongs to the Section Mycotoxins)
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13 pages, 1449 KiB  
Article
Novel DNA Barcoding and Multiplex PCR Strategy for the Molecular Identification and Mycotoxin Gene Detection of Fusarium spp. in Maize from Bulgaria
by Daniela Stoeva, Deyana Gencheva, Georgi Radoslavov, Peter Hristov, Rozalina Yordanova and Georgi Beev
Methods Protoc. 2025, 8(4), 78; https://doi.org/10.3390/mps8040078 - 9 Jul 2025
Viewed by 322
Abstract
Fusarium spp. represent a critical threat to maize production and food safety due to their mycotoxin production. This study introduces a refined molecular identification protocol integrating four genomic regions—ITS1, IGS, TEF-1α, and β-TUB—for robust species differentiation of Fusarium spp. isolates from [...] Read more.
Fusarium spp. represent a critical threat to maize production and food safety due to their mycotoxin production. This study introduces a refined molecular identification protocol integrating four genomic regions—ITS1, IGS, TEF-1α, and β-TUB—for robust species differentiation of Fusarium spp. isolates from post-harvest maize in Bulgaria. The protocol enhances species resolution, especially for closely related taxa within the Fusarium fujikuroi species complex (FFSC). A newly optimized multiplex PCR strategy was developed using three primer sets, each designed to co-amplify a specific pair of toxigenic genes: fum6/fum8, tri5/tri6, and tri5/zea2. Although all five genes were analyzed, they were detected through separate two-target reactions, not in a single multiplex tube. Among 17 identified isolates, F. proliferatum (52.9%) dominated, followed by F. verticillioides, F. oxysporum, F. fujikuroi, and F. subglutinans. All isolates harbored at least one toxin biosynthesis gene, with 18% co-harboring genes for both fumonisins and zearalenone. This dual-protocol approach enhances diagnostic precision and supports targeted mycotoxin risk management strategies. Full article
(This article belongs to the Section Molecular and Cellular Biology)
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17 pages, 1765 KiB  
Article
Multi-Mycotoxin Analyses by UPLC-MS/MS in Wheat: The Situation in Belgium in 2023 and 2024
by Camille Jonard, Anne Chandelier, Damien Eylenbosch, Joke Pannecoucque, Bruno Godin, Caroline Douny, Marie-Louise Scippo and Sébastien Gofflot
Foods 2025, 14(13), 2300; https://doi.org/10.3390/foods14132300 - 28 Jun 2025
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Abstract
This work proposes an insight into the mycotoxins detected in wheat from the 2023 and 2024 harvests in Belgium and highlights the link between agronomic conditions and mycotoxin contamination. The study utilized samples from a Belgian trial network, covering nine locations in 2023 [...] Read more.
This work proposes an insight into the mycotoxins detected in wheat from the 2023 and 2024 harvests in Belgium and highlights the link between agronomic conditions and mycotoxin contamination. The study utilized samples from a Belgian trial network, covering nine locations in 2023 and eight in 2024, ensuring diverse pedoclimatic contexts and including 11 different varieties. Sowing and harvest dates, previous crops and meteorological data were collected for these locations. A validated UPLC-MS/MS multi-mycotoxin method able to detect 20 mycotoxins, regulated or not, was used. Deoxynivalenol, zearalenone, and enniatins B and B1 were detected in the 2023 and 2024 samples. Enniatin A1 was only detected in the 2024 samples. Mycotoxin contamination was higher in 2024 compared to 2023, in terms of both the number of contaminated samples and the contamination levels. Enniatins B and B1, non-regulated mycotoxins, were widely detected in the 2024 wheat samples, with enniatin B detected in 68 out 88 samples ranging from 12 to 488 µg/kg. Differences between the wheat varieties were observed, with some varieties showing significantly higher contamination. Additionally, geographic location appeared to influence contamination levels, which could be related to previous crops or meteorological events. In conclusion, this research provides a comprehensive analysis of mycotoxin co-contamination in wheat samples from diverse pedoclimatic contexts in Belgium based over 2 years. It shows the importance of weather conditions on mycotoxin contamination. It also emphasizes the importance of variety selection to manage mycotoxin contamination. Full article
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