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Mycotoxins Study: Toxicology, Identification and Control

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

Deadline for manuscript submissions: closed (30 November 2020) | Viewed by 42409

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Prof. Dr. Cristina Juan García

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Department of Preventive Medicine and Public Health, Food Sciences, Forensic Medicine and Toxicology, University of Valencia | UV, E-46100 Valencia, Spain
Interests: food safety; mass spectrometry analysis; proteomics; risk assessment; toxicology
Special Issues, Collections and Topics in MDPI journals

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Dear Colleagues,

The evaluation of the presence of mycotoxins in different matrices is achieved through different analytical tools (including quantitative or qualitative determinations). Studies of mycotoxins used QuEChERS, solid–liquid, liquid–liquid, and solid-phase extraction or inmunoaffinity colums for micotoxins isolation, in combination with chromatographyc equipments (GC or LC) coupled to spectrometry detectors (QTrap-MS/MS, MS/MS tandem, QTOF-MS/MS). All these studies represent key steps in the establishment of the limits of detection, limits of quantification, points of identification, accuracy, reproducibility, and/or repeatability of different procedures. The maximum permitted or recommended levels for mycotoxins in different matrices are comprised within a wide range (including the levels tolerated by infants and animals). In addition, the climatic changes influence the growth of fungi in different matrices, therefore, their control and evaluation are demanded by authorities and safety food systems.

These authorities are concerned not only with the determination of mycotoxins presence but also with the toxicological effects of mycotoxins, and in vivo or in vitro assays are necessary for a complete evaluation. In fact, these assays are the basis for the control and prevention of population exposure to mycotoxins in dietary exposure studies. The last surveys focused on regulated mycotoxins (aflatoxins, fumonisins, and trichothecenes) and emerging toxins such as enniatins and beauvericin in adult consumers, while very few studies have monitored mycotoxins levels in infant products.

The focus of this Special Issue of Toxins is to gather the most recent reports on the toxicological effects of single or combined mycotoxins (genotoxicity, neurotoxicity, or metabolomics effects), the identification of known and unknown mycotoxins, masked mycotoxins, and biometabolized mycotoxins in different matrices (including food, feed, and biological samples), and the development of analytical skills to control their presence. Research papers and review articles describing novelties or overviews, respectively, are welcome.

Prof. Dr. Cristina Juan García
Guest Editor

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Keywords

mycotoxins
in vivo
in vitro
metabolomics
food safety
mycotoxin analysis
genotoxicity
neurotoxicity

Published Papers (13 papers)

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Editorial

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3 pages, 213 KiB

Open AccessEditorial

Mycotoxins: Toxicology, Identification and Control

by Cristina Juan García

Toxins 2021, 13(4), 242; https://doi.org/10.3390/toxins13040242 - 29 Mar 2021

Cited by 5 | Viewed by 1711

Abstract

The evaluation of the presence of mycotoxins in different matrices is achieved through different analytical tools (including quantitative or qualitative determinations) [...] Full article

(This article belongs to the Special Issue Mycotoxins Study: Toxicology, Identification and Control)

Research

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15 pages, 561 KiB

Open AccessArticle

Occurrence of Free and Conjugated Mycotoxins in Aromatic and Medicinal Plants and Dietary Exposure Assessment in the Moroccan Population

by Aicha El Jai, Abdellah Zinedine, Ana Juan-García, Jordi Mañes, Samira Etahiri and Cristina Juan

Toxins 2021, 13(2), 125; https://doi.org/10.3390/toxins13020125 - 08 Feb 2021

Cited by 14 | Viewed by 2796

Abstract

Aromatic and medicinal plants (AMPs), as herbal material, are subjected to contamination by various mycotoxin-producing fungi, either free and conjugated. Such a problem is associated with poor storage practices, and lack of adopting good agricultural practices and good harvesting practices. Nevertheless, AMPs are poorly investigated. The purpose of this study was to investigate the co-occurrence of 15 mycotoxins (four aflatoxins (AFB1, AFB2, AFG1, and AFG2), ochratoxin A (OTA), beauvericin (BEA), four enniatins (ENA, ENA1, ENB, and ENB1), zearalenone (ZEN), alternariol (AOH), tentoxin (TENT), T-2, and HT-2 toxins) in 40 samples of AMPs frequently consumed in Morocco by using liquid chromatography tandem mass spectrometry. Evaluation of conjugated mycotoxins and their identification using liquid chromatography coupled to time-of-flight mass spectrometry with ion mass exact was also carried out. Results showed that 90% of the analyzed samples presented at least one mycotoxin, and 52% presented co-occurrence of them. Mycotoxins detected were: AOH (85%), ZEN (27.5%), β-ZEL (22%), AFG1 (17.5%), TENT (17.5%), ENB (10%), AFG2 (7.5%), α-ZEL (5%), ENA1 (2.5%), and HT-2 (2.5%), while the conjugated mycotoxins were ZEN-14-Glc (11%) and ZEN-14-Sulf (9%). The highest observed level was for AOH, with 309 ng/g. Ten samples exceeded the recommended levels set by the European Pharmacopoeia for AF mycotoxins in plant material (4 ng/g), and three samples exceeded the maximum limits for AFs (10 ng/g) in species established by the European Commission. Although the co-occurrence of several mycotoxins in AMP samples was observed, the dietary exposure assessment showed that the intake of mycotoxins through the consumption of AMP beverages does not represent a risk for the population. Full article

(This article belongs to the Special Issue Mycotoxins Study: Toxicology, Identification and Control)

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12 pages, 1479 KiB

Open AccessArticle

Using Cholinesterases and Immobilized Luminescent Photobacteria for the Express-Analysis of Mycotoxins and Estimating the Efficiency of Their Enzymatic Hydrolysis

by Elena Efremenko, Olga Maslova, Nikolay Stepanov and Anvar Ismailov

Toxins 2021, 13(1), 34; https://doi.org/10.3390/toxins13010034 - 06 Jan 2021

Cited by 7 | Viewed by 1998

Abstract

Novel sensitive analytical agents that can be used for simple, affordable, and rapid analysis of mycotoxins are urgently needed in scientific practice, especially for the screening of perspective bio-destructors of the toxic contaminants. We compared the characteristics of a rapid quantitative analysis of different mycotoxins (deoxynivalenol, ochratoxin A, patulin, sterigmatocystin, and zearalenone) using acetyl-, butyrylcholinesterases and photobacterial strains of luminescent cells in the current study. The best bioindicators in terms of sensitivity and working range (μg/mL) were determined as follows: Photobacterium sp. 17 cells for analysis of deoxynivalenol (0.8–89) and patulin (0.2–32); Photobacterium sp. 9.2 cells for analysis of ochratoxin A (0.4–72) and zearalenone (0.2–32); acetylcholinesterase for analysis of sterigmatocystin (0.12–219). The cells were found to be more sensitive than enzymes. The assayed strains of photobacterial cells ensured 44%–83% lower limit of detection for deoxynivalenol and sterigmatocystin as compared to the previously known data for immobilized luminescent cells, and the range of working concentrations was extended by a factor of 1.5–3.5. Calibration curves for the quantitative determination of patulin using immobilized photobacteria were presented in this work for the first time. This calibration was applied to estimate the enzyme efficiency for hydrolyzing mycotoxins using zearalenone and His₆-tagged organophosphorus hydrolase as examples. Full article

(This article belongs to the Special Issue Mycotoxins Study: Toxicology, Identification and Control)

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9 pages, 932 KiB

Open AccessArticle

Efficacy of Potentially Probiotic Fruit-Derived Lactobacillus fermentum, L. paracasei and L. plantarum to Remove Aflatoxin M₁ In Vitro

by Paloma Oliveira da Cruz, Clarisse Jales de Matos, Yuri Mangueira Nascimento, Josean Fechine Tavares, Evandro Leite de Souza and Hemerson Iury Ferreira Magalhães

Toxins 2021, 13(1), 4; https://doi.org/10.3390/toxins13010004 - 23 Dec 2020

Cited by 12 | Viewed by 5287

Abstract

This study evaluated the efficacy of potentially probiotic fruit-derived Lactobacillus isolates, namely, L. paracasei 108, L. plantarum 49, and L. fermentum 111, to remove aflatoxin M₁ (AFM₁) from a phosphate buffer solution (PBS; spiked with 0.15 µg/mL AFM₁). The efficacy of examined isolates (approximately 10⁹ cfu/mL) as viable and non-viable cells (heat-killed; 100 °C, 1 h) to remove AFM₁ was measured after 1 and 24 h at 37 °C. The recovery of AFM₁ bound to bacterial cells after washing with PBS was also evaluated. Levels of AFM₁ in PBS were measured with high-performance liquid chromatography. Viable and non-viable cells of all examined isolates were capable of removing AFM₁ in PBS with removal percentage values in the range of 73.9–80.0% and 72.9–78.7%, respectively. Viable and non-viable cells of all examined Lactobacillus isolates had similar abilities to remove AFM₁. Only L. paracasei 108 showed higher values of AFM₁ removal after 24 h for both viable and non-viable cells. Percentage values of recovered AFM₁ from viable and non-viable cells after washing were in the range of 13.4–60.6% and 10.9–47.9%, respectively. L. plantarum 49 showed the highest AFM₁ retention capacity after washing. L. paracasei 108, L. plantarum 49, and L. fermentum 111 could have potential application to reduce AFM₁ to safe levels in foods and feeds. The cell viability of examined isolates was not a pre-requisite for their capacity to remove and retain AFM₁. Full article

(This article belongs to the Special Issue Mycotoxins Study: Toxicology, Identification and Control)

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14 pages, 2293 KiB

Open AccessArticle

Acetamiprid Affects Destruxins Production but Its Accumulation in Metarhizium sp. Spores Increases Infection Ability of Fungi

by Monika Nowak, Przemysław Bernat, Julia Mrozińska and Sylwia Różalska

Toxins 2020, 12(9), 587; https://doi.org/10.3390/toxins12090587 - 11 Sep 2020

Cited by 8 | Viewed by 2584

Abstract

Metarhizium sp. are entomopathogenic fungi that inhabit the soil environment. Together, they act as natural pest control factors. In the natural environment, they come into contact with various anthropogenic pollutants, and sometimes, they are used together and interchangeably with chemical insecticides (e.g., neonicotinoids) for pest control. In most cases, the compatibility of entomopathogens with insecticides has been determined; however, the influence of these compounds on the metabolism of entomopathogenic fungi has not yet been studied. Secondary metabolites are very important factors that influence the fitness of the producers, playing important roles in the ability of these pathogens to successfully parasitize insects. In this study, for the first time, we focus on whether the insecticide present in the fungal growth environment affects secondary metabolism in fungi. The research revealed that acetamiprid at concentrations from 5 to 50 mg L⁻¹ did not inhibit the growth of all tested Metarhizium sp.; however, it reduced the level of 19 produced destruxins in direct proportion to the dosage used. Furthermore, it was shown that acetamiprid accumulates not only in plant or animal tissues, but also in fungal cells. Despite the negative impact of acetamiprid on secondary metabolism, it was proofed to accumulate in Metarhizium spores, which appeared to have a stronger infectious potential against mealworm Tenebrio molitor, in comparison to the insecticide or the biological agent alone. Full article

(This article belongs to the Special Issue Mycotoxins Study: Toxicology, Identification and Control)

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11 pages, 752 KiB

Open AccessArticle

Biomonitoring of Enniatin B1 and Its Phase I Metabolites in Human Urine: First Large-Scale Study

by Yelko Rodríguez-Carrasco, Alfonso Narváez, Luana Izzo, Anna Gaspari, Giulia Graziani and Alberto Ritieni

Toxins 2020, 12(6), 415; https://doi.org/10.3390/toxins12060415 - 22 Jun 2020

Cited by 14 | Viewed by 2594

Abstract

Enniatins (Enns) are mycotoxins produced by Fusarium spp. which are a fungus widely spread throughout cereals and cereal-based products. Among all the identified enniatins, Enn B1 stands as one of the most prevalent analogues in cereals in Europe. Hence, the aim of this study was to evaluate for the first time the presence of Enn B1 and its phase I metabolites in 300 human urine samples using an ultrahigh-performance liquid chromatography high resolution mass spectrometry (UHPLC-Q-Orbitrap HRMS) methodology. Enn B1 was detected in 94.3% of samples ranging from 0.007 to 0.429 ng/mL (mean value: 0.065 ng/mL). In accordance with previous in vitro and in vivo analysis, hydroxylated metabolites (78.0% samples) and carbonylated metabolites (66.0% samples) were tentatively identified as the major products. Results from this biomonitoring study point to a frequent intake of Enn B1 in the studied population, suggesting that in-depth toxicological studies are needed in order to understand the potential effects in humans. Full article

(This article belongs to the Special Issue Mycotoxins Study: Toxicology, Identification and Control)

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10 pages, 300 KiB

Open AccessArticle

Ochratoxin A in Beers Marketed in Portugal: Occurrence and Human Risk Assessment

by Liliana J. G. Silva, Ana C. Teixeira, André M. P. T. Pereira, Angelina Pena and Celeste M. Lino

Toxins 2020, 12(4), 249; https://doi.org/10.3390/toxins12040249 - 12 Apr 2020

Cited by 8 | Viewed by 2600

Abstract

Ochratoxin A (OTA) is produced by fungi present in several agricultural products with much relevance to food safety. Since this mycotoxin is widely found in cereals, beer has a potential contamination risk. Therefore, it was deemed essential to quantify, for the first time, the levels of OTA in beer, a cereal-based product that is marketed in Portugal, as well as to calculate the human estimated weekly intake (EWI) and risk assessment. A total of 85 samples were analyzed through immunoaffinity clean-up, followed by liquid chromatography-fluorescence detection (LC-FD). This analytical methodology allowed a limit of quantification (LOQ) of 0.43 µg/L. The results showed that 10.6% were contaminated at levels ranging between <LOQ and 11.25 µg/L, with an average of 3.14 ± 4.09 µg/L. Samples of industrial production presented lower incidence and contamination levels than homemade and craft beers. On what concerns human risk, the calculated EWI was significantly lower than the tolerable weekly intake (TWI). However, in the worst case scenario, based on a high concentration, the rate EWI/TWI was 138.01%. Full article

(This article belongs to the Special Issue Mycotoxins Study: Toxicology, Identification and Control)

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16 pages, 2963 KiB

Open AccessEditor’s ChoiceArticle

Individual and Combined Effect of Zearalenone Derivates and Beauvericin Mycotoxins on SH-SY5Y Cells

by Fojan Agahi, Guillermina Font, Cristina Juan and Ana Juan-García

Toxins 2020, 12(4), 212; https://doi.org/10.3390/toxins12040212 - 27 Mar 2020

Cited by 30 | Viewed by 3464

Abstract

Beauvericin (BEA) and zearalenone derivatives, α-zearalenol (α-ZEL), and β-zearalenol (β-ZEL), are produced by several Fusarium species. Considering the impact of various mycotoxins on human’s health, this study determined and evaluated the cytotoxic effect of individual, binary, and tertiary mycotoxin treatments consisting of α-ZEL, β-ZEL, and BEA at different concentrations over 24, 48, and 72 h on SH-SY5Y neuronal cells, by using the MTT assay (3-(4,5-dimethylthiazol-2-yl)-2,5diphenyltetrazoliumbromide). Subsequently, the isobologram method was applied to elucidate if the mixtures produced synergism, antagonism, or additive effects. Ultimately, we determined the amount of mycotoxin recovered from the media after treatment using liquid chromatography coupled with electrospray ionization–quadrupole time-of-flight mass spectrometry (LC–ESI–qTOF-MS). The IC₅₀ values detected at all assayed times ranged from 95 to 0.2 μM for the individual treatments. The result indicated that β-ZEL was the most cytotoxic mycotoxin when tested individually. The major effect detected for all combinations assayed was synergism. Among the combinations assayed, α-ZEL + β-ZEL + BEA and α-ZEL + BEA presented the highest cytotoxic potential with respect to the IC value. In individual treatment, α-ZEL was the most recovered mycotoxin; while, this was observed for BEA in binary combination α-ZEL + BEA. Full article

(This article belongs to the Special Issue Mycotoxins Study: Toxicology, Identification and Control)

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10 pages, 2100 KiB

Open AccessEditor’s ChoiceArticle

Effect of Ozone and Electron Beam Irradiation on Degradation of Zearalenone and Ochratoxin A

by Kai Yang, Ke Li, Lihong Pan, Xiaohu Luo, Jiali Xing, Jing Wang, Li Wang, Ren Wang, Yuheng Zhai and Zhengxing Chen

Toxins 2020, 12(2), 138; https://doi.org/10.3390/toxins12020138 - 24 Feb 2020

Cited by 33 | Viewed by 3205

Abstract

Zearalenone (ZEN) and ochratoxin A (OTA) are key concerns of the food industry because of their toxicity and pollution scope. This study investigated the effects of ozone and electron beam irradiation (EBI) on the degradation of ZEN and OTA. Results demonstrated that 2 mL of 50 μg/mL ZEN was completely degraded after 10 s of treatment by 2.0 mg/L ozone. The degradation rate of 1 μg/mL ZEN by 16 kGy EBI was 92.76%. Methanol was superior to acetonitrile in terms of degrading ZEN when the irradiation dose was higher than 6 kGy. The degradation rate of 2 mL of 5 μg/mL OTA by 50 mg/L ozone at 180 s was 34%, and that of 1 μg/mL OTA by 16 kGy EBI exceeded 90%. Moreover, OTA degraded more rapidly in acetonitrile. Ozone performed better in the degradation of ZEN, whereas EBI was better for OTA. The conclusions provide theoretical and practical bases for the degradation of different fungal toxins. Full article

(This article belongs to the Special Issue Mycotoxins Study: Toxicology, Identification and Control)

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10 pages, 24186 KiB

Open AccessArticle

Interactions of Destruxin A with Silkworms’ Arginine tRNA Synthetase and Lamin-C Proteins

by Jingjing Wang, Qunfang Weng, Fei Yin and Qiongbo Hu

Toxins 2020, 12(2), 137; https://doi.org/10.3390/toxins12020137 - 22 Feb 2020

Cited by 10 | Viewed by 2749

Abstract

Destruxin A (DA), a cyclodepsipeptidic mycotoxin produced by entomopathogenic fungus Metarhizium anisopliae, has good insecticidal activity and potential to be a new pesticide. However, the mechanism of action is still obscure. Our previous experiments showed that DA was involved in regulation of transcription and protein synthesis and suggested that silkworms’ arginine tRNA synthetase (BmArgRS), Lamin-C Proteins (BmLamin-C) and ATP-dependent RNA helicase PRP1 (BmPRP1) were candidates of DA-binding proteins. In this study, we employed bio-layer interferometry (BLI), circular dichroism (CD), cellular thermal shift assay (CETSA), and other technologies to verify the interaction of DA with above three proteins in vitro and in vivo. The results of BLI indicated that BmArgRS and BmLamin-C were binding-protein of DA with K_D value 5.53 × 10⁻⁵ and 8.64 × 10⁻⁵ M, but not BmPRP1. These interactions were also verified by CD and CETSA tests. In addition, docking model and mutants assay in vitro showed that BmArgRS interacts with DA at the pocket including Lys228, His231, Asp434 and Gln437 in its enzyme active catalysis region, while BmLamin-C binds to DA at His524 and Lys528 in the tail domain. This study might provide new insight and evidence in illustrating molecular mechanism of DA in breaking insect. Full article

(This article belongs to the Special Issue Mycotoxins Study: Toxicology, Identification and Control)

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14 pages, 1330 KiB

Open AccessArticle

Efficient and Simultaneous Chitosan-Mediated Removal of 11 Mycotoxins from Palm Kernel Cake

by Atena Abbasi Pirouz, Jinap Selamat, Shahzad Zafar Iqbal and Nik Iskandar Putra Samsudin

Toxins 2020, 12(2), 115; https://doi.org/10.3390/toxins12020115 - 12 Feb 2020

Cited by 17 | Viewed by 3407

Abstract

Mycotoxins are an important class of pollutants that are toxic and hazardous to animal and human health. Consequently, various methods have been explored to abate their effects, among which adsorbent has found prominent application. Liquid chromatography tandem mass spectrometry (LC–MS/MS) has recently been applied for the concurrent evaluation of multiple mycotoxins. This study investigated the optimization of the simultaneous removal of mycotoxins in palm kernel cake (PKC) using chitosan. The removal of 11 mycotoxins such as aflatoxins (AFB₁, AFB₂, AFG₁ and AFG₂), ochratoxin A (OTA), zearalenone (ZEA), fumonisins (FB1 and FB2) and trichothecenes (deoxynivalenol (DON), HT-2 and T-2 toxin) from palm kernel cake (PKC) was studied. The effects of operating parameters such as pH (3–6), temperature (30–50 °C) and time (4–8 h) on the removal of the mycotoxins were investigated using response surface methodology (RSM). Response surface models obtained with R² values ranging from 0.89–0.98 fitted well with the experimental data, except for the trichothecenes. The optimum point was obtained at pH 4, 8 h and 35 °C. The maximum removal achieved with chitosan for AFB₁, AFB₂, AFG₁, AFG₂, OTA, ZEA, FB₁ and FB₂ under the optimized conditions were 94.35, 45.90, 82.11, 84.29, 90.03, 51.30, 90.53 and 90.18%, respectively. Full article

(This article belongs to the Special Issue Mycotoxins Study: Toxicology, Identification and Control)

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10 pages, 817 KiB

Open AccessEditor’s ChoiceArticle

Patulin Mycotoxin in Mango and Orange Fruits, Juices, Pulps, and Jams Marketed in Pakistan

by Shabbir Hussain, Muhammad Rafique Asi, Mazhar Iqbal, Nisha Khalid, Syed Wajih-ul-Hassan and Agustín Ariño

Toxins 2020, 12(1), 52; https://doi.org/10.3390/toxins12010052 - 16 Jan 2020

Cited by 31 | Viewed by 5025

Abstract

The objective of the study was to explore the incidence of patulin (PAT) mycotoxin in mango and orange fruits and derived products marketed in Pakistan. A total of 274 samples, including 70 mango fruits, 63 mango-based products (juices, pulp, and jam), 77 orange fruits, and 64 orange-based products, were collected. PAT was determined by reverse-phase high-performance liquid chromatography (HPLC) with UV-Vis detector (276 nm). Linear detector response was observed (R² > 0.99), the limit of detection (LOD) was 5 µg/kg and recovery percentage was 97.4%. The incidence of PAT in mango samples was 61.7%, and the concentration ranged from <LOD to 6415 µg/kg with a mean of 110.9 µg/kg. Our results showed the high susceptibility of mango fruits to patulin, and it was observed that decayed mango fruits were most contaminated with PAT. Among the mango samples, PAT concentration was higher in fruits than in processed products such as mango juice, pulp, and jam. Toxin incidence in orange samples was 52.5% with concentrations from <LOD to 61 µg/kg and a mean of 6.3 µg/kg. As much as 29 samples of mango (21.8%) contained PAT concentration above the regulatory limit (50 µg/kg), whereas there was only one exceeding orange sample (0.7%). Our results show that PAT seems to be a problem in fruits, juices, and derived solid products, especially from mango, and needs surveillance on regular basis. Full article

(This article belongs to the Special Issue Mycotoxins Study: Toxicology, Identification and Control)

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Review

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16 pages, 1323 KiB

Open AccessReview

Recent Advances on Macrocyclic Trichothecenes, Their Bioactivities and Biosynthetic Pathway

by Muzi Zhu, Youfei Cen, Wei Ye, Saini Li and Weimin Zhang

Toxins 2020, 12(6), 417; https://doi.org/10.3390/toxins12060417 - 23 Jun 2020

Cited by 18 | Viewed by 3170

Abstract

Macrocyclic trichothecenes are an important group of trichothecenes bearing a large ring. Despite the fact that many of trichothecenes are of concern in agriculture, food contamination, health care and building protection, the macrocyclic ones are becoming the research hotspot because of their diversity in structure and biologic activity. Several researchers have declared that macrocyclic trichothecenes have great potential to be developed as antitumor agents, due to the plenty of their compounds and bioactivities. In this review we summarize the newly discovered macrocyclic trichothecenes and their bioactivities over the last decade, as well as identifications of genes tri17 and tri18 involved in the trichothecene biosynthesis and putative biosynthetic pathway. According to the search results in database and phylogenetic trees generated in the review, the species of the genera Podostroma and Monosporascus would probably be great sources for producing macrocyclic trichothecenes. Moreover, we propose that the macrocyclic trichothecene roridin E could be formed via acylation or esterification of the long side chain linked with C-4 to the hydroxyl group at C-15, and vice versa. More assays and evidences are needed to support this hypothesis, which would promote the verification of the proposed pathway. Full article

(This article belongs to the Special Issue Mycotoxins Study: Toxicology, Identification and Control)

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