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Novel Approaches in Mycotoxins Research: Detection, Prevention and Mode of Action

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A special issue of Toxins (ISSN 2072-6651). This special issue belongs to the section "Mycotoxins".

Deadline for manuscript submissions: 28 February 2025 | Viewed by 15009

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Special Issue Editors

Dr. Daniela Jakšić

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Guest Editor

Department of Microbiology, Faculty of Pharmacy and Biochemistry, University of Zagreb, Zagreb, Croatia
Interests: toxins

Dr. Ana Juan-García

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Guest Editor

Department of Preventive Medicine and Public Health, Food Sciences, Forensic Medicine and Toxicology, University of Valencia | UV, E-46100 Valencia, Spain
Interests: food safety; toxicology; in vitro studies; risk asessment
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Special Issue Information

Dear Colleagues,

Mycotoxins are a chemically diverse group of fungal secondary metabolites that are toxic to humans when ingested, inhaled, or absorbed into the skin. They may be detected in various food and feed matrices, but also in environmental samples. Despite the long history of mycotoxins research, there is a gap of knowledge about human exposure to certain groups of mycotoxins. Most mycotoxins are thoroughly studied in vitro and in vivo, but the models that are used do not fully reflect their action in humans. To support the clinical relevance of mycotoxins, it is necessary to provide a supported association of mycotoxins to certain clinical pictures. Quintessential to this are the data on the presence of mycotoxins in human biological fluids and tissues, exposure assessment studies, and modes of actions in advanced animal-free systems, such as human cell lines, co-cultures, and organoids. Additionally, for well-known mycotoxins, efficient and adequate methods are urgently needed for their elimination from the food chain and indoor environments.

Thus, the aim of this Special Issue is to motivate researchers and gather information focused on the following topics:

Human exposure to mycotoxins:

Detection of the mycotoxins in biological fluids and tissues;
Biomarkers for exposure to mycotoxins.

Control and elimination of mycotoxins:

Inhibitors of fungal growth or mycotoxin producing abilities;
Physicochemical binders or adsorbents of mycotoxins;
Degradation of mycotoxins by plants, fungi, bacteria, and/or their products.

Mechanisms of action of mycotoxins in advanced animal-free systems:

Human cell lines, co-cultures;
Organoids

Review articles about these topics.

Dr. Daniela Jakšić
Dr. Ana Juan-García
Guest Editors

Manuscript Submission Information

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Published Papers (9 papers)

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Research

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15 pages, 1340 KiB

Open AccessArticle

Inhibition of Aflatoxin B1 Production by Procyanidins Present in Annona muricata and Uncaria tomentosa Aqueous Extracts

by Laura F. Cadenillas, Guillaume Billerach, Christopher Hernandez, Vanessa Durrieu and Jean-Denis Bailly

Toxins 2024, 16(11), 454; https://doi.org/10.3390/toxins16110454 - 23 Oct 2024

Viewed by 530

Abstract

Aflatoxin B₁ (AFB₁), primarily produced by Aspergillus flavus and A. parasiticus, is the most dangerous mycotoxin for humans and contaminates a variety of crops. To limit fungal growth and aflatoxin production in food and feed, research has been increasingly focusing on alternatives to pesticides. Studies show that some aqueous plant extracts with strong antioxidant properties could significantly impact AFB₁ production, representing an eco-friendly and sustainable method to protect crops. The present study demonstrates that aqueous extracts of Anonna muricata (AM) and Uncaria tomentosa (UT) inhibit AFB₁ synthesis in a dose-dependent manner with a half-maximal inhibitory concentration of 0.25 and 0.28 mg dry matter per milliliter of culture medium, respectively. This effect correlates with the presence of polyphenols and, more precisely, with condensed tannins. It is also related to the subsequent antioxidant activity of both extracts. A bio-guided fractionation followed by high-performance liquid chromatography and mass spectrometry analysis of the active fractions identifies procyanidins and, more precisely, catechin (5.3% w/w for AM and 5.4% w/w for UT) and epicatechin (10.6% w/w for AM and 25.7% w/w for UT) as the major components in both extracts. The analysis of how pure standards of these molecules affect AFB₁ production demonstrates that catechin plays an essential role in the inhibition observed for both plant extracts, since the pure standard inhibits 45% of AFB₁ synthesis at a concentration close to that of the extracts. Full article

(This article belongs to the Special Issue Novel Approaches in Mycotoxins Research: Detection, Prevention and Mode of Action)

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14 pages, 2458 KiB

Open AccessArticle

First Report on Mycotoxin Contamination of Hops (Humulus lupulus L.)

by Ivana Dodlek Šarkanj, Nada Vahčić, Ksenija Markov, Josip Haramija, Natalija Uršulin-Trstenjak, Krunoslav Hajdek, Michael Sulyok, Rudolf Krska and Bojan Šarkanj

Toxins 2024, 16(7), 293; https://doi.org/10.3390/toxins16070293 - 26 Jun 2024

Viewed by 1379

Abstract

The presence of mycotoxins and other toxic metabolites in hops (Humulus lupulus L.) was assessed for the first time. In total, 62 hop samples were sampled in craft breweries, and analyzed by a multi-toxin LS-MS/MS method. The study collected samples from craft breweries in all of the Croatian counties and statistically compared the results. Based on previous reports on Alternaria spp. and Fusarium spp. contamination of hops, the study confirmed the contamination of hops with these toxins. Alternaria toxins, particularly tenuazonic acid, were found in all tested samples, while Fusarium toxins, including deoxynivalenol, were present in 98% of samples. However, no Aspergillus or Penicillium metabolites were detected, indicating proper storage conditions. In addition to the Alternaria and Fusarium toxins, abscisic acid, a drought stress indicator in hops, was also detected, as well as several unspecific metabolites. The findings suggest the need for monitoring, risk assessment, and potential regulation of Alternaria and Fusarium toxins in hops to ensure the safety of hop usage in the brewing and pharmaceutical industries. Also, four local wild varieties were tested, with similar results to the commercial varieties for toxin contamination, but the statistically significant regional differences in toxin occurrence highlight the importance and need for targeted monitoring. Full article

(This article belongs to the Special Issue Novel Approaches in Mycotoxins Research: Detection, Prevention and Mode of Action)

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13 pages, 25063 KiB

Open AccessArticle

Mechanism of Fumonisin Self-Resistance: Fusarium verticillioides Contains Four Fumonisin B₁-Insensitive-Ceramide Synthases

by Tamara Krska, Krisztian Twaruschek, Gerlinde Wiesenberger, Franz Berthiller and Gerhard Adam

Toxins 2024, 16(6), 235; https://doi.org/10.3390/toxins16060235 - 22 May 2024

Viewed by 988

Abstract

Fusarium verticillioides produces fumonisins, which are mycotoxins inhibiting sphingolipid biosynthesis in humans, animals, and other eukaryotes. Fumonisins are presumed virulence factors of plant pathogens, but may also play a role in interactions between competing fungi. We observed higher resistance to added fumonisin B₁ (FB₁) in fumonisin-producing Fusarium verticillioides than in nonproducing F. graminearum, and likewise between isolates of Aspergillus and Alternaria differing in production of sphinganine-analog toxins. It has been reported that in F. verticillioides, ceramide synthase encoded in the fumonisin biosynthetic gene cluster is responsible for self-resistance. We reinvestigated the role of FUM17 and FUM18 by generating a double mutant strain in a fum1 background. Nearly unchanged resistance to added FB₁ was observed compared to the parental fum1 strain. A recently developed fumonisin-sensitive baker’s yeast strain allowed for the testing of candidate ceramide synthases by heterologous expression. The overexpression of the yeast LAC1 gene, but not LAG1, increased fumonisin resistance. High-level resistance was conferred by FUM18, but not by FUM17. Likewise, strong resistance to FB₁ was caused by overexpression of the presumed F. verticillioides “housekeeping” ceramide synthases CER1, CER2, and CER3, located outside the fumonisin cluster, indicating that F. verticillioides possesses a redundant set of insensitive targets as a self-resistance mechanism. Full article

(This article belongs to the Special Issue Novel Approaches in Mycotoxins Research: Detection, Prevention and Mode of Action)

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17 pages, 1122 KiB

Open AccessArticle

High-Performance Liquid Chromatography–Fluorescence Detection Method for Ochratoxin A Quantification in Small Mice Sample Volumes: Versatile Application across Diverse Matrices Relevant for Neurodegeneration Research

by Elba Beraza, Maria Serrano-Civantos, Maria Izco, Lydia Alvarez-Erviti, Elena Gonzalez-Peñas and Ariane Vettorazzi

Toxins 2024, 16(5), 213; https://doi.org/10.3390/toxins16050213 - 3 May 2024

Viewed by 1169

Abstract

Ochratoxin A (OTA) is a mycotoxin commonly found in various food products, which poses potential health risks to humans and animals. Recently, more attention has been directed towards its potential neurodegenerative effects. However, there are currently no fully validated HPLC analytical methods established for its quantification in mice, the primary animal model in this field, that include pivotal tissues in this area of research, such as the intestine and brain. To address this gap, we developed and validated a highly sensitive, rapid, and simple method using HPLC-FLD for OTA determination in mice tissues (kidney, liver, brain, and intestine) as well as plasma samples. The method was rigorously validated for selectivity, linearity, accuracy, precision, recovery, dilution integrity, carry-over effect, stability, and robustness, meeting the validation criteria outlined by FDA and EMA guidelines. Furthermore, the described method enables the quantification of OTA in each individual sample using minimal tissue mass while maintaining excellent recovery values. The applicability of the method was demonstrated in a repeated low-dose OTA study in Balb/c mice, which, together with the inclusion of relevant and less common tissues in the validation process, underscore its suitability for neurodegeneration-related research. Full article

(This article belongs to the Special Issue Novel Approaches in Mycotoxins Research: Detection, Prevention and Mode of Action)

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18 pages, 1960 KiB

Open AccessArticle

In Vitro Digestion and Intestinal Absorption of Mycotoxins Due to Exposure from Breakfast Cereals: Implications for Children’s Health

by Soraia V. M. de Sá, Miguel A. Faria, José O. Fernandes and Sara C. Cunha

Toxins 2024, 16(5), 205; https://doi.org/10.3390/toxins16050205 - 25 Apr 2024

Cited by 1 | Viewed by 1799

Abstract

Breakfast cereals play a crucial role in children’s diets, providing essential nutrients that are vital for their growth and development. Children are known to be more susceptible than adults to the harmful effects of food contaminants, with mycotoxins being a common concern in cereals. This study specifically investigated aflatoxin B1 (AFB1), enniatin B (ENNB), and sterigmatocystin (STG), three well-characterized mycotoxins found in cereals. The research aimed to address existing knowledge gaps by comprehensively evaluating the bioaccessibility and intestinal absorption of these three mycotoxins, both individually and in combination, when consumed with breakfast cereals and milk. The in vitro gastrointestinal method revealed patterns in the bioaccessibility of AFB1, ENNB, and STG. Overall, bioaccessibility increased as the food progressed from the stomach to the intestinal compartment, with the exception of ENNB, whose behavior differed depending on the type of milk. The ranking of overall bioaccessibility in different matrices was as follows: digested cereal > cereal with semi-skimmed milk > cereal with lactose-free milk > cereal with soy beverage. Bioaccessibility percentages varied considerably, ranging from 3.1% to 86.2% for AFB1, 1.5% to 59.3% for STG, and 0.6% to 98.2% for ENNB. Overall, the inclusion of milk in the ingested mixture had a greater impact on bioaccessibility compared to consuming the mycotoxins as a single compound or in combination. During intestinal transport, ENNB and STG exhibited the highest absorption rates when ingested together. This study highlights the importance of investigating the combined ingestion and transport of these mycotoxins to comprehensively assess their absorption and potential toxicity in humans, considering their frequent co-occurrence and the possibility of simultaneous exposure. Full article

(This article belongs to the Special Issue Novel Approaches in Mycotoxins Research: Detection, Prevention and Mode of Action)

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26 pages, 3495 KiB

Open AccessArticle

Discovering the Protective Effects of Quercetin on Aflatoxin B1-Induced Toxicity in Bovine Foetal Hepatocyte-Derived Cells (BFH12)

by Marianna Pauletto, Mery Giantin, Roberta Tolosi, Irene Bassan, Anisa Bardhi, Andrea Barbarossa, Ludovica Montanucci, Anna Zaghini and Mauro Dacasto

Toxins 2023, 15(9), 555; https://doi.org/10.3390/toxins15090555 - 6 Sep 2023

Cited by 5 | Viewed by 2143

Abstract

Aflatoxin B1 (AFB1) induces lipid peroxidation and mortality in bovine foetal hepatocyte-derived cells (BFH12), with underlying transcriptional perturbations associated mainly with cancer, cellular damage, inflammation, bioactivation, and detoxification pathways. In this cell line, curcumin and resveratrol have proven to be effective in mitigating AFB1-induced toxicity. In this paper, we preliminarily assessed the potential anti-AFB1 activity of a natural polyphenol, quercetin (QUE), in BFH12 cells. To this end, we primarily measured QUE cytotoxicity using a WST-1 reagent. Then, we pre-treated the cells with QUE and exposed them to AFB1. The protective role of QUE was evaluated by measuring cytotoxicity, transcriptional changes (RNA-sequencing), lipid peroxidation (malondialdehyde production), and targeted post-transcriptional modifications (NQO1 and CYP3A enzymatic activity). The results demonstrated that QUE, like curcumin and resveratrol, reduced AFB1-induced cytotoxicity and lipid peroxidation and caused larger transcriptional variations than AFB1 alone. Most of the differentially expressed genes were involved in lipid homeostasis, inflammatory and immune processes, and carcinogenesis. As for enzymatic activities, QUE significantly reverted CYP3A variations induced by AFB1, but not those of NQO1. This study provides new knowledge about key molecular mechanisms involved in QUE-mediated protection against AFB1 toxicity and encourages in vivo studies to assess QUE’s bioavailability and beneficial effects on aflatoxicosis. Full article

(This article belongs to the Special Issue Novel Approaches in Mycotoxins Research: Detection, Prevention and Mode of Action)

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11 pages, 2673 KiB

Open AccessArticle

The Impact of Storage Temperature and Time on Ergot Alkaloid Concentrations

by Jensen E. Cherewyk, Taylor J. Grusie-Ogilvie, Sarah E. Parker, Barry R. Blakley and Ahmad N. Al-Dissi

Toxins 2023, 15(8), 497; https://doi.org/10.3390/toxins15080497 - 5 Aug 2023

Cited by 2 | Viewed by 1618

Abstract

Ergot sclerotia produce toxic secondary metabolites, ergot alkaloids, that infect cereal crops and grasses. Ergot alkaloids have two isomeric configurations: the C-8-R-isomer (R-epimer), and the C-8-S-isomer (S-epimer). Ergot contaminated matrices, such as cereal grains or grasses, may be stored for extended periods at various temperatures before being analyzed, utilized, or consumed. This study assessed the concentration of six common ergot alkaloids in both configurations found in naturally contaminated wheat over time (one, two, and four months) at different temperatures (room temperature, +4 °C, and −20 °C) using ultra-high-performance liquid chromatography–tandem mass spectrometry. The data indicate that the total ergot concentration within a natural contaminated sample varies over time at room temperature, +4 °C, and −20 °C. The total ergot concentration increased until month two, and decreased at month four, independent of temperature (p < 0.05). The total R-epimer concentration appeared to be less stable over time than the total S-epimer concentration. The changes in the total R and total S-epimer concentrations may have been caused by changes in the ergocristine and ergocristinine concentrations, respectively. Time and temperature should be considered when storing potentially contaminated matrices in a laboratory or practical agriculture situations. Quantification of ergot contaminated matrices should occur prior to their use to ensure the most reliable estimates of the concentration of ergot. Full article

(This article belongs to the Special Issue Novel Approaches in Mycotoxins Research: Detection, Prevention and Mode of Action)

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Review

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30 pages, 910 KiB

Open AccessReview

Effect of Acrylamide and Mycotoxins in SH-SY5Y Cells: A Review

by Luna Bridgeman, Cristina Juan, Houda Berrada and Ana Juan-García

Toxins 2024, 16(2), 87; https://doi.org/10.3390/toxins16020087 - 6 Feb 2024

Viewed by 2407

Abstract

Thermal processes induce the formation of undesired toxic components, such as acrylamide (AA), which has been shown to induce brain toxicity in humans and classified as Group 2A by the International Agency of Research in Cancer (IARC), as well as some mycotoxins. AA and mycotoxins’ toxicity is studied in several in vitro models, including the neuroblastoma cell line model SH-SY5Y cells. Both AA and mycotoxins occur together in the same food matrix cereal base (bread, pasta, potatoes, coffee roasting, etc.). Therefore, the goal of this review is to deepen the knowledge about the neurological effects that AA and mycotoxins can induce on the in vitro model SH-SY5Y and its mechanism of action (MoA) focusing on the experimental assays reported in publications of the last 10 years. The analysis of the latest publications shows that most of them are focused on cytotoxicity, apoptosis, and alteration in protein expression, while others are interested in oxidative stress, axonopathy, and the disruption of neurite outgrowth. While both AA and mycotoxins have been studied in SH-SY5Y cells separately, the mixture of them is starting to draw the interest of the scientific community. This highlights a new and interesting field to explore due to the findings reported in several publications that can be compared and the implications in human health that both could cause. In relation to the assays used, the most employed were the MTT, axonopathy, and qPCR assays. The concentration dose range studied was 0.1–10 mM for AA and 2 fM to 200 µM depending on the toxicity and time of exposure for mycotoxins. A healthy and varied diet allows the incorporation of a large family of bioactive compounds that can mitigate the toxic effects associated with contaminants present in food. Although this has been reported in some publications for mycotoxins, there is still a big gap for AA which evidences that more investigations are needed to better explore the risks for human health when exposed to AA and mycotoxins. Full article

(This article belongs to the Special Issue Novel Approaches in Mycotoxins Research: Detection, Prevention and Mode of Action)

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30 pages, 3531 KiB

Open AccessSystematic Review

Machine Learning Applied to the Detection of Mycotoxin in Food: A Systematic Review

by Alan Inglis, Andrew C. Parnell, Natarajan Subramani and Fiona M. Doohan

Toxins 2024, 16(6), 268; https://doi.org/10.3390/toxins16060268 - 12 Jun 2024

Cited by 3 | Viewed by 2095

Abstract

Mycotoxins, toxic secondary metabolites produced by certain fungi, pose significant threats to global food safety and public health. These compounds can contaminate a variety of crops, leading to economic losses and health risks to both humans and animals. Traditional lab analysis methods for mycotoxin detection can be time-consuming and may not always be suitable for large-scale screenings. However, in recent years, machine learning (ML) methods have gained popularity for use in the detection of mycotoxins and in the food safety industry in general due to their accurate and timely predictions. We provide a systematic review on some of the recent ML applications for detecting/predicting the presence of mycotoxin on a variety of food ingredients, highlighting their advantages, challenges, and potential for future advancements. We address the need for reproducibility and transparency in ML research through open access to data and code. An observation from our findings is the frequent lack of detailed reporting on hyperparameters in many studies and a lack of open source code, which raises concerns about the reproducibility and optimisation of the ML models used. The findings reveal that while the majority of studies predominantly utilised neural networks for mycotoxin detection, there was a notable diversity in the types of neural network architectures employed, with convolutional neural networks being the most popular. Full article

(This article belongs to the Special Issue Novel Approaches in Mycotoxins Research: Detection, Prevention and Mode of Action)

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