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Pesticides and mycotoxins, prominent chemical hazards in the food chain, are commonly found in plant-based foods, contributing to their pervasive presence in the human body, as evidenced by biomonitoring programs. Despite this, there is limited knowledge about their co-occurrence patterns. While intervention studies have demonstrated that organic diets can significantly reduce pesticide levels, their impact on mycotoxin exposure has been overlooked. To address this gap, this study pursued two objectives: first, to characterize the simultaneous presence of mycotoxins and pesticides in human urine samples by means of the control of the biomarkers of exposure, and second, to investigate the influence of consuming organic foods on these co-exposure patterns. A pilot study involving 20 healthy volunteers was conducted, with participants consuming either exclusively organic or conventional foods during a 24-h diet intervention in autumn 2021 and spring 2022 to account for seasonal variability. Participants provided detailed 24-h dietary records, and their first-morning urine samples were collected, minimally treated and analysed using LC-Q-ToF-MS by means of a multitargeted method in order to detect the presence of these residues. Results indicated that among the 52 screened compounds, four mycotoxins and seven pesticides were detected in over 25% of the samples. Deoxynivalenol (DON) and the non-specific pesticide metabolite diethylphosphate (DEP) exhibited the highest frequency rates (100%) and concentration levels. Correlations were observed between urine levels of mycotoxins (DON, ochratoxin alpha [OTα], and enniatin B [ENNB]) and organophosphate pesticide metabolites DEP and 2-diethylamino-6-methyl-4-pyrimidinol (DEAMPY). The pilot intervention study suggested a reduction in ENNB and OTα levels and an increase in β-zearalenol levels in urine after a short-term replacement with organic food. However, caution is advised due to the study's small sample size and short duration, emphasizing the need for further research to enhance understanding of the human chemical exposome and refine chemical risk assessment., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier Ltd. All rights reserved.)

Enniatin B (ENNB) is a mycotoxin produced by moulds from the Fusarium genera and its toxic effects are still not fully elucidated, hence a safe reference exposure value has not been established yet. ENNB is the most prevalent emerging mycotoxin and is widely found in cereal-based products, nevertheless, there are no comprehensive exposure assessment studies. For that reason, the aim of this study was to characterise the occurrence of ENNB and estimate the exposure of the Spanish and Catalan populations. A total of 347 cereal-based products were collected in 2019 and were analysed using liquid chromatography-tandem mass spectrometry. Consumption data were obtained from the national food consumption surveys (ENALIA) and a regional survey conducted in Catalonia. The global exposure was estimated using deterministic and probabilistic methods. The results showed a high occurrence of close to 100% in all foodstuffs, with a range from 6 to 269 µg/kg, and a strong correlation with the levels of deoxynivalenol. Children aged one-nine years were the most exposed, showing mean estimates in the range 308-324 ng/kg bw/day and 95th percentiles 697-781 ng/kg bw/day. This study stresses the need for further toxicological data to establish reference doses and conclude formal risk assessment, accounting for the co-occurrence with deoxynivalenol.

Agronomic factors can affect mycotoxin contamination of maize, one of the most produced cereals. Maize is usually harvested at 18% moisture, but it is not microbiologically stable until it reaches 14% moisture at the drying plants. We studied how three agronomic factors (crop diversification, tillage system and nitrogen fertilization rate) can affect fungal and mycotoxin contamination (deoxynivalenol and fumonisins B 1 and B 2 ) in maize at harvest. In addition, changes in maize during a simulated harvest-till-drying period were studied. DON content at harvest was higher for maize under intensive tillage than using direct drilling (2695 and 474 μg kg -1 , respectively). We found two reasons for this: (i) soil crusting in intensive tillage plots caused the formation of pools of water that created high air humidity conditions, favouring the development of DON-producing moulds; (ii) the population of Lumbricus terrestris , an earthworm that would indirectly minimize fungal infection and mycotoxin production on maize kernels, is reduced in intensive tillage plots. Therefore, direct drilling is a better approach than intensive tillage for both preventing DON contamination and preserving soil quality. Concerning the simulated harvest-till-drying period, DON significantly increased between storage days 0 and 5. Water activity dropped on the 4th day, below the threshold for DON production (around 0.91). From our perspective, this study constitutes a step forward towards understanding the relationships between agronomic factors and mycotoxin contamination in maize, and towards improving food safety.

Deoxynivalenol (DON) remains one of the most concerning mycotoxins produced by the Fusarium genus due to the wide occurrence in highly consumed cereal-based food and its associated toxicological effects. Previous studies conducted in Spain and other European countries suggested that some vulnerable groups such as children could be exceeding the tolerable daily intakes. Thus, the aim of this study was to conduct a comprehensive and updated dietary exposure assessment study in Spain, with a specific analysis in the region of Catalonia. Cereal-based food samples collected during 2019 were analysed using liquid chromatography coupled to tandem mass spectrometry for multi-mycotoxin detection including DON and its main metabolites and derivatives. Consumption data were gathered from the nation-wide food surveys ENALIA and ENALIA2 conducted in Spain, and a specific survey conducted in Catalonia. The data were combined using deterministic and semi-parametric probabilistic methods. The results showed that DON was widely present in cereal-based food highly consumed in Spain and the Catalonia region. Exposure to DON among the adult population was globally low; however, among infants aged 3-9 years, it resulted in the median of 192 ng/kg body weight/day and the 95th percentiles of 604 ng/kg body weight/day, that would exceed the most conservative safety threshold for infants. Bread and pasta were the main contributing foodstuffs to the global exposure to DON, even among infants; thus, those foods should be considered a priority for food control or to develop strategies to reduce the exposure. In any case, further toxicological and epidemiological studies are required in order to refine the safety thresholds accounting for the sensitivity of the infant population.

In 2009 the EU Regulation 386/2009 established a new functional group of feed additives called "substances for reduction of the contamination of feed by mycotoxin". Later, di-octahedral bentonite (1 m558) was authorised, as an anti-aflatoxin additive, being the only additive of this group authorised to date. This work aims to demonstrate the effectiveness of other bentonites, such as tri-octahedral bentonites, versus Fusarium- mycotoxins, since very few adsorbents have proved their effectiveness in relation to this group of mycotoxins. For this purpose, 7 bentonites (six of them tri-octahedral) and 7 commercial adsorbents, added at 0.02% (w/v), were assayed in an in vitro adsorption experiment using two simulated gastro-intestinal (GI) juices (pH 1.3 and 6.8) versus zearalenone (ZEN: 0.1-5 mg/L), fumonisin B 1 (FB 1 : 1-10 mg/L) and deoxynivalenol (DON: 2-10 mg/L). Mycotoxin adsorption data were fitted to Langmuir and Freundlich isotherms. In vitro adsorption experiments showed that ZEN and FB 1 (in the latter case only in acid medium) were partially adsorbed, while the adsorption of DON was negligible. Moreover, the increase of adsorbent dose (up to 0.20%, w/v) significantly improved the in vitro adsorption of ZEN and FB 1 , reaching >90% of adsorption. The present work proposes the use of some tri-octahedral bentonites as feed additives for Fusarium- mycotoxin reduction.

Mycotoxins produced by certain fungal species of the Fusarium genus are frequently found as contaminants in cereals and feedstuffs. Fumonisins (FBs), deoxynivalenol (DON) and zearalenone (ZEN) are of special concern relative to animal health and productivity. The aim of this work was to analyse the levels of Fusarium mycotoxin contamination in samples of total mixed rations (TMRs) for dairy cows. To accomplish this analysis, an HPLC-MS/MS multi-mycotoxin method was developed and validated. The relation between the formulation of TMR samples and the presence of mycotoxins was also studied. From February 2016 to January 2018, a total of 193 TMR samples for dairy cows collected from farms located in different areas of Spain were analysed for the presence of FBs, ZEN, DON and their metabolites. In total, 112 samples (58%) were contaminated with at least one mycotoxin, and 38 samples (20%) presented more than one mycotoxin. FBs were the mycotoxins most frequently found (34% positive samples). DON was detected in 17% of samples, and ZEN was detected in 16% of samples. Among the metabolites analysed, only deoxynivalenol-3-glucoside (DON-3-Glc) and 15-acetyldeoxynivalenol (15-ADON) were detected. The levels of all the Fusarium mycotoxins studied were always below the values recommended by the European Commission for feedstuffs. The wide variety of ingredients used in the formulation of the analysed samples made it difficult to reach definite conclusions, although it seemed that some cereal silages and concentrates such as cereals or compound feed used as ingredients of the TMR may be related to the presence of mycotoxins.

The aim of this study was to evaluate the effect of 15 commercial yeasts in the mitigation of the Fusarium mycotoxins deoxynivalenol (DON) and zearalenone (ZEN) during the brewing process. Saccharomyces strains (10 strains of S. cerevisiae and 5 of S. pastorianus) were used to ferment DON and ZEN contaminated wort. Wort samples were taken every 24 h during fermentation, while mycotoxin analysis in yeast was performed at the end of fermentation (96 h); additionally, pH and ethanol content were measured daily. For mycotoxin analysis, after immunoaffinity purification of sample extracts, analysis was performed using an Ultra-High-Pressure Liquid Chromatograph coupled with a diode array or fluorescence detector (UHPLC-DAD/FLD). Mycotoxin presence had no significant effect on the ethanol production during brewing. At the end of fermentation, 10-17% of DON and 30-70% of ZEN had been removed, 6% of the initial concentration of DON and 31% of the ZEN being adsorbed by the yeast. Beermakers must pay careful attention to the raw material since a high percentage of DON could be present at the end of the beer fermentation process. Future studies should focus on the quantification of "masked" mycotoxins that are relevant to food security., (Copyright © 2019 Elsevier Ltd. All rights reserved.)

The aim of the present study was to evaluate the occurrence of 23 mycotoxins in beer purchased in Mexico and to assess two exposure scenarios in the Mexican population through beer consumption. Multi-mycotoxin analysis of a total of 61 different beers (132 samples) was carried out using UHPLC-MS/MS equipment. Probability density functions were used to describe mycotoxins contamination. The daily intake of mycotoxins was estimated using a semi-probabilistic approach, applying the Monte Carlo method. Deoxynivalenol (DON) and its metabolites (deoxynivalenol-3-glucoside (DON3G) and 3-acetyl-deoxynivalenol (3ADON)) were the mycotoxins found in higher proportions in contaminated samples. None of the other mycotoxins overpassed the limit of quantification (LOQ) of the method. The combined intake of DON and its analogues ranged from 5.24 to 86.59 ng kg -1  bw day -1 , which represent from 1.20 to 19.83% of the DON TDI. The results suggest that depending on the individual consumption of beer and depending on the type of beer, the intake of DON via beer could represent a significant percentage of the tolerable daily intake (TDI).

The fate of deoxynivalenol, deoxynivalenol-3-glucoside, 3- and 15-acetyl-deoxynivalenol, zearalenone, α- and β-zearalenol and fumonisins (fumonisin B 1 and fumonisin B 2 ) through mashing and wort boiling was studied. Three different mycotoxin contamination scenarios were considered. In almost all samples an increase in the level of mycotoxins in wort was observed during mashing followed by a decrease after just 30 min of the process, with levels remaining constant until the end of boiling. Deoxynivalenol and its metabolites were reduced to their initial level contained in the malt before mashing, or even lower, however in none of the samples they were completely eliminated. Zearalenone was not quantitated at the end of boiling, although there was a significant initial level of ZEN. β-Zearalenol remained unaltered during the process. Fumonisins were reduced by between 50 and 100 per cent during mashing and boiling., (Copyright © 2018 Elsevier Ltd. All rights reserved.)

Mycotoxins are secondary metabolites produced by fungi that may contaminate different foods intended for human consumption, resulting in a widespread exposure worldwide. The novel exposome paradigm has the ambition to decipher the different environmental insults threating human health throughout the entire lifespan. Given the large potential impact of mycotoxins in terms of human exposure and related health effects, the ambition of this review is to present this group of chemical compounds and the high interest to be included in exposome projects. Furthermore, we also attempt to approach the novel exposome paradigm to more traditional disciplines such as mycotoxin exposure assessment and mycotoxicology, introducing the new methodological challenges and translational needs. Hence, we provide an overview of major biomarkers currently developed, biological matrices where these may be found, an overview of internal exposure levels and potential co-occurrence with environmental chemicals and finally an overview of major health effects with the illustrative example of the potent xenoestrogen zearalenol. Conversely, these new approaches may be an excellent opportunity to fill many research gaps on mycotoxins research as the identification of associations with human health, elucidation of join effect with other environmental exposures or the decipher of underlying molecular mechanisms by using advanced OMICs technologies., (Copyright © 2018 Elsevier Ltd. All rights reserved.)

Intraspecies variability in fungal growth and mycotoxin production has important implications for food safety. Using the Bioscreen C we have examined spectrophotometrically intraspecies variability of A. flavus using 10 isolates under different environments, including temperature shifts, in terms of growth and aflatoxin B 1 (AFB 1 ) production. Five high and five low AFB 1 producers were examined. The study was conducted at 5 isothermal conditions (from 15 to 37 °C) and 4 dynamic scenarios (between 15 and 30 °C). The experiments were carried out in a semisolid YES medium at 0.92 a w and two inoculum levels, 10 2 and 10 3  spores/mL. The Time to Detection (TTD) of growth initiation was determined and modelled as a function of temperature through a polynomial equation and the model was used to predict TTD under temperature upshifts conditions using a novel approach. The results obtained in this study have shown that a model can be developed to describe the effect of temperature upshifts on the TTD for all the studied isolates and inoculum levels. Isolate variability increased as the growth conditions became more stressful and with a lower inoculum level. Inoculum level affected the intraspecies variability but not the repeatability of the experiments. In dynamic conditions, isolate responses depended both on the temperature shift and, predominantly, the final temperature level. AFB 1 production was highly variable among the isolates and greatly depended on temperature (optimum temperature at 30-35 °C) and inoculum levels, with often higher production with lower inoculum. This suggests that, from an ecological point of view, the potential isolate variability and interaction with dynamic conditions should be taken into account in developing strategies to control growth and predicting mycotoxin risks by mycotoxigenic fungi., (Copyright © 2018 Elsevier B.V. All rights reserved.)

The major objective of this study was to describe the effect of water activity and temperature on radial growth and production of ochratoxin A (OTA) and zearalenone (ZEA) on sorghum grains of three Aspergillus tubingensis and three Fusarium incarnatum isolates. The water activity range was 0.91-0.99 a w for F. incarnatum isolates and 0.88-0.99 a w for A. tubingensis isolates. Temperatures of incubation were 15, 25 and 37°C for both species. Mycotoxin production was determined after 7, 14, 21 and 28days depending on the growth rate of the six isolates. Maximum growth rates (mm/day) were observed at 37°C and 0.99 a w for A. tubingensis isolates and at 0.99 a w and 25°C for F. incarnatum isolates. A. tubingensis was able to grow at 15°C only at the highest a w levels (0.97 and 0.99 a w ). However, at this temperature F. incarnatum grew at 0.94 a w . Optimum ochratoxin A production was observed at 0.97 a w ×37°C whereas optimal conditions for ZEA production varied from one isolate to another. Moreover, isolates of F. incarnatum from Tunisia do not require high a w and temperature levels to yield maximum levels of ZEA. In general, our results showed that there is no correlation between the growth and production of ZEA in the case of F. incarnatum. This is the first study on the water activity and temperature effect on growth rate and ZEA production of F. incarnatum. Our results show that sorghum grains not only support growth but also OTA and ZEA production by A. tubingensis and F. incarnatum, respectively., (Copyright © 2016 Elsevier B.V. All rights reserved.)

The stability of deoxynivalenol (DON), deoxynivalenol-3-glucoside (DON-3-glucoside), 3-acetyldeoxynivalenol (3-ADON), 15-acetyldeoxynivalenol (15-ADON), de-epoxy-deoxynivalenol (DOM-1) and ochratoxin A (OTA) during thermal processing has been studied. Baking temperature, time and initial mycotoxin concentration in the raw materials were assayed as factors. An improved UPLC-MS/MS method to detect DON, DON-3-glucoside, 3-ADON, 15-ADON and DOM-1 in wheat baked products was developed in the present assay. The results highlighted the importance of temperature and time in mycotoxin stability in heat treatments. OTA is more stable than DON in a baking treatment. Interestingly, the DON-3-glucoside concentrations increased (>300%) under mild baking conditions. On the other hand, it was rapidly reduced under harsh conditions. The 3-ADON decreased during the heat treatment; while DOM-1 increased after the heating process. Finally, the data followed first order kinetics for analysed mycotoxins and thermal constant rates (k) were calculated. This parameter can be a useful tool for prediction of mycotoxin levels., (Copyright © 2015 Elsevier Ltd. All rights reserved.)

The effects of two exposure times per day (6 and 16 h) of UV-A or UV-B radiation, combined with dark and dark plus light incubation periods during 7-21 d on fungal growth and mycotoxins production of Aspergillus species were studied. Aspergillus carbonarius and Aspergillus parasiticus were inoculated on grape and pistachio media under diurnal and nocturnal temperatures choosing light photoperiod according to harvest conditions of these crops in Spain. Ultraviolet irradiation had a significant effect on A. carbonarius and A. parasiticus colony size (diameter, biomass dry weight, and colony density) and mycotoxin accumulation, although intraspecies differences were observed. Inhibition of A. carbonarius fungal growth decreased when exposure time was reduced from 16 h to 6 h, but this was not always true for ochratoxin A (OTA) production. OTA reduction was higher under UV-A than UV-B radiation and the reduction increased along time conversely to the aflatoxins (AFs). Aflatoxin B1 (AFB1) was the main toxin produced by A. parasiticus except in the UV-B light irradiated colonies which showed a higher percentage of AFG than AFB. Morphological changes were observed in colonies grown under UV-B light., (Copyright © 2014 The British Mycological Society. Published by Elsevier Ltd. All rights reserved.)

The goals of the present work were, on the one hand, to assess the effect of baking on the stability of zearalenone (ZEA) and deoxynivalenol (DON), as well as the transfer of DON from pasta to boiling water, and, on the other hand, to quantify the impact of DON depletion, during cooking of pasta, on overall exposure estimates. Therefore, the bread-making process was simulated on a pilot-plant scale by using naturally contaminated flour with DON and ZEA. Transfer of DON from pasta to water was evaluated at different boiling times. Pasta was prepared on a pilot-plant scale by using naturally contaminated durum wheat flour; subsequently, it was boiled simulating home cooking. The experiments examined the stability of DON and ZEA during the bread-making process, including fermentation with Saccharomyces cerevisiae and baking at 200°C. Our results showed a high transfer of DON from pasta to boiling water, reaching depletion levels of almost 75%, which correlated with levels found in water. Accordingly, these cooking depletion rates were computed through a stochastic exposure model to weight their impact on the final exposure estimates. Finally, statistically significant differences were found in most of the parameters and populations assessed, but these were not enough to consider the process as protective because the contribution of pasta to the overall DON intake was commonly low.

Background: Fusarium graminearum and F. verticillioides are two very important mycotoxigenic species as they cause diverse diseases in crops. The effects of constant and cycling temperatures on growth and mycotoxin production of these species were studied on soybean based medium and on irradiated soya beans., Results: F. graminearum grew better when was incubated at 15, 20 and 15-20 °C (isothermal or cycling temperature) during 21 days of incubation. Maximum levels of zearalenone and deoxynivalenol (39.25 and 1040.4 µg g(-1), respectively) were detected on soya beans after 15 days of incubation and the optimal temperature for mycotoxin production was 15 °C for zearalenone and 20 °C for deoxynivalenol. F. verticillioides grew better at 25 °C in culture medium and at 15/20 °C and 15/25 °C on soybean seeds. Fumonisin B(1) was produced only in culture medium, and the maximum level (7.38 µg g(-1)) was found at 15 °C after 7 days of incubation., Conclusion: When growth and mycotoxin production under cycling temperatures were predicted from the results under constant conditions, observed values were different from calculated for both species and substrate medium. Therefore, care should be taken if data at constant temperature conditions are to be extrapolated to real field conditions., (Copyright © 2012 Society of Chemical Industry.)

Cereals are a very important part of the human and animal diets. However, agricultural products can be contaminated by moulds and their mycotoxins. Plant extracts, particularly those of Equisetum arvense and Stevia rebaudiana have been reported previously to contain antioxidant compounds which may have antifungal properties. In this study, E. arvense and S. rebaudiana extracts were tested for their control of mycotoxigenic fungi in maize. The extracts were tested separately and as a mixture for their effect on growth of Aspergillus flavus and Fusarium verticillioides. Extracts were added to unsterilised inoculated maize at different water activity (a(w)) levels (0.85-0.95). Moulds were inoculated and incubated for 30 days. Results confirmed that the extract of E. arvense and a mixture 1:1 of Equisetum-Stevia may be effective for the inhibition of both growth of A. flavus and aflatoxin production at high water activity levels (pre-harvest conditions). In general, growth of the F. verticillioides was reduced by the use of plant extracts, especially at 0.95 a(w). However, fumonisin presence was not significantly affected. E. arvense and S. rebaudiana extracts could be developed as an alternative treatment to control aflatoxigenic mycobiota in moist maize., (Copyright © 2011 Elsevier B.V. All rights reserved.)

The need to ensure the microbiological quality and safety of food products has stimulated interest in the use of mathematical models for quantifying and predicting microbial behaviour. For 20 years, predictive microbiology has been developed for predicting the occurrence of food-borne pathogens, although these tools are dedicated to bacteria. Recently, the situation has changed and a growing number of studies are available in the literature dealing with the predictive modelling approach of fungi. To our knowledge the present one is the first review focussed on predictive mycology and food safety, including mycotoxins; existing kinetic and probability models applied to mycotoxigenic fungi germination and growth, and mycotoxin production are reviewed.

Eleven fungi isolated from grapes and sun-dried grapes (Alternaria alternata, Cladosporium herbarum, Eurotium amstelodami, Penicillium janthinellum, P. decumbens, Trichoderma harzianum, Candida sp., Aspergillus carbonarius OTA-negative, A. carbonarius OTA-positive, A. niger var. niger. and A. japonicus var. aculeatus), were grown in SNM medium at different water activities (0.82-0.97) and temperature (20-40 degrees C) levels for 18 days. Pairs of one Aspergillus faced with one non-Aspergillus were grown at 0.87-0.97 a(w) and at 20 and 30 degrees C. In single cultures, daily radii were recorded. In paired cultures radii were recorded and each A. section Nigri isolate was given a dominance score. At high temperatures and low water activities, Penicillium isolates, E. amstelodami and A. niger var. niger showed higher growth rates, while T. harzianum only grew well at the highest water activity. In addition, A. section Nigri was dominant in most paired assays, being only surpassed by T. harzianum at 0.97 a(w) and 20 degrees C. Thus, prevalence of A. section Nigri in sun-dried grapes can be explained by its adaptation to environmental conditions of sun-drying, and by its capability to dominate other fungal species when coming into contact with them.

Contamination of cereals with mycotoxigenic species of Fusarium is an important source of trichothecenes, fumonisins and other mycotoxins which cause serious diseases in human and animals. In addition, these species are phytopathogenic and produce severe losses in cereal yield. Methods for early detection of these Fusarium species are crucial to prevent toxins entering the food chain and are a useful tool in disease management practices. We have developed an integrated protocol for diagnosis of mycotoxigenic Fusarium contamination in maize which can also be used for other cereals. The protocol consisted in an easy and rapid DNA extraction from maize samples (grain and germ), and subsequent group-specific polymerase chain reaction (PCR) assays for genus Fusarium, Gibberella fujikuroi complex, and trichothecene-producing species of Fusarium, that orientate the search of the critical species. We have additionally developed a PCR assay for the identification of F. proliferatum. The primers were designed on the basis of IGS sequence (Intergenic Spacer of rDNA), a multi-copy region in the genome that permits to enhance the sensitivity of the assay in comparison with PCR assays based on single-copy sequences. The suitability of the protocol and the relative efficacy of single and multi-copy sequence-based PCR assays have been tested in a wide range of fumonisin-contaminated maize samples.

The mycotoxin deoxynivalenol (DON) is one of the most common mycotoxins of cereals worldwide, and its occurrence has been widely reported in raw wheat. The free mycotoxin form is not the only route of exposure; modified forms can also be present in cereal products. Deoxynivalenol-3-glucoside (DON-3-glucoside) is a common DON plant conjugate. The mycotoxin concentration could be affected by food processing; here, we studied the stability of DON and DON-3-glucoside during baking of small doughs made from white wheat flour and other ingredients. A range of common food additives and ingredients were added to assess possible interference: ascorbic acid (E300), citric acid (E330), sorbic acid (E200), calcium propionate (E282), lecithin (E322), diacetyltartaric acid esters of fatty acid mono- and diglycerides (E472a), calcium phosphate (E341), disodium diphosphate (E450i), xanthan gum (E415), polydextrose (E1200), sorbitol (E420i), sodium bicarbonate (E500i), wheat gluten and malt flour. The DON content was reduced by 40%, and the DON-3-glucoside concentration increased by >100%, after baking for 20 min at 180°C. This confirmed that DON and DON-3-glucoside concentrations can vary during heating, and DON-3-glucoside could even increase after baking. However, DON and DON-3-glucoside are not affected significantly by the presence of the food additives tested. [ABSTRACT FROM AUTHOR]

Mycotoxins are secondary fungal metabolites of high concern in the food and feed industry. Their presence in many cereal-based products has been numerously reported. Beer is the most consumed alcoholic beverage worldwide, and Fusarium mycotoxins originating from the malted and unmalted cereals might reach the final product. This review aims to describe the possible Fusarium fungi that could infect the cereals used in beer production, the transfer of mycotoxins throughout malting and brewing as well as an insight into the incidence of mycotoxins in the craft beer segment of the industry. Studies show that germination is the malting step that can lead to a significant increase in the level of all Fusarium mycotoxins. The first step of mashing (45 °C) has been proved to possess the most significant impact in the transfer of hydrophilic toxins from the grist into the wort. However, during fermentation, a slight reduction of deoxynivalenol, and especially of zearalenone, is achieved. This review also highlights the limited research available on craft beer and the occurrence of mycotoxins in these products. [ABSTRACT FROM AUTHOR]

Abstract: Penicillium expansum is the most important fungal species causing spoilage in cold stored apples. P. expansum produces patulin, a toxic secondary metabolite. The aim of this work was to study the changes in growth parameters and patulin accumulation that may occur when two different isolates of P. expansum germinate and develop in the same wound. The effect of the inoculum size was also studied. For that purpose, apples were point inoculated with two different isolates of P. expansum at two different conidia concentrations. Both isolates were also inoculated together. Apples were cold stored at 1°C or incubated at 20°C until lesion diameter reached 2.5cm. Patulin from decayed tissue was analysed at the end of cold storage/incubation. It seems that intraspecific interactions occur. The interactions resulted in slower growth in which patulin content was lower. A competition for resources followed by either a “combat-like” interaction may be involved in this. Higher inoculum size led to shorter lag phase, which reflects the importance of the hygiene in packinghouses facilities. The inoculum size also affected growth rate and patulin accumulation, which suggests that interactions between individuals of the same strain may occur. Competition for resources and a combat-like interaction may take place. In vitro studies should be carried out in order to assess the patterns of the interactions observed. [Copyright &y& Elsevier]

The article explores the reduction of aflatoxin (AF) B1, B2, G1, and G2 as a function of initial moisture content of samples, barrel temperature, and residence time when artificially contaminated rice meal was extrusion-cooked. Extrusion-cooking was noted to reduce AF content. Only in the case of AFG2 was it found that the higher the temperature, the higher the moisture content, and the longer the residence time, the greater the reduction.

This review focuses on the fumonisin-producing Fusarium species and the ecophysiology of these species. The effects of environmental biotic and abiotic factors on germination, growth, and fumonisin B1 production by Fusarium verticillioides and Fusarium proliferatum have been investigated under laboratory, field, and storage conditions. An understanding of the factors involved in production of fumonisins is the first step in preventing accumulation of these toxins. [ABSTRACT FROM AUTHOR]

Aflatoxins are the most potent genotoxic and carcinogenic mycotoxins. To date, research has only focused on the presence of free aflatoxins in agricultural commodities. Therefore, the main objective of this study was to investigate the occurrence of possible modified aflatoxins in maize. Different hydrolysis methods were applied to convert modified mycotoxins into their free aflatoxins. Eighteen aflatoxin-contaminated maize samples were incubated with potassium hydroxide, trifluoromethanesulfonic acid and several enzymes to induce hydrolysis. Potassium hydroxide caused a total reduction of aflatoxins, while trifluoromethanesulfonic acid did not lead to an increase in free aflatoxins, neither did treatment with a protease. However, α-amylase and cellulase incubation caused significant increases in the total free aflatoxin content, 15 ± 8% and 13 ± 5%, respectively. These results show that a small proportion of aflatoxins could be associated to matrix substances in plants. Consequently, hydrolysis could occur during food processing and during mammalian digestion, leading to an underestimation of the total aflatoxin content. [ABSTRACT FROM AUTHOR]

Abstract: Predictive mycology has dealt mainly with germination, growth and inactivation of fungi while the issue of mycotoxin production remains relatively unexplored. Very few studies provide biomass dry weight/colony size data along with mycotoxin data for the same sample times, thus the ratio mycotoxin accumulation per fungal biomass dry weight/colony size has rarely been reported. For this reason, the objective of the present study was to model the kinetics of mycotoxin production under the assumption of existing both no-growth-associated and growth-associated production. Aspergillus flavus was chosen as a model mycotoxigenic microorganism, and it was grown in maize agar medium and maize grain at 0.90 and 0.99 aw at 25°C. A significant positive correlation (p<0.05) was observed among the biomass responses (colony radius and biomass dry weight) in agar medium and colony radius in maize at both a w levels assayed. The Luedeking–Piret model was used to model AFB1 production and reasonable percentages of variability were explained. Moreover, AFB1 production was in general slightly better predicted through colony area. As conclusion, aflatoxin production may follow a mixed-growth associated trend, confirming that toxin formation does not present a clear delay in relation to growth under certain conditions. [Copyright &y& Elsevier]

Abstract: The aim of this work was to model the growth of Aspergillus parasiticus and Aspergillus ochraceus, both mycotoxin producers, near to the growth/no growth boundaries and validate those models in sterile maize grain, peanuts and coffee beans. Malt extract agar was adjusted to six different water activities: 0.93, 0.91, 0.89, 0.87, 0.85 and 0.80. Plates were incubated at 10, 15, 20, 25, 30, 37 and 42°C. For each of the 42 conditions, 10 Petri dishes were inoculated. Both kinetic and probability models were applied to colony growth data. The results of the present study indicate that the developed probability modelling approach could be satisfactorily employed to quantify the combined effect of temperature and water activity on the growth responses of A. ochraceus and A. parasiticus. However, validation of kinetic results led to poor goodness of prediction. In this study, the validation samples were placed near to the expected boundaries of the models in order to test them under the worst situation. Probability of growth prediction under extreme growth conditions was somewhat compromised, but it can be considered acceptable. [Copyright &y& Elsevier]

Abstract: The aim of this study was to assess the impact of suboptimal environmental conditions on the intraspecific variability of A. carbonarius growth and OTA production using thirty isolates of A. carbonarius. Three aw/temperature conditions were tested, one optimal (0.98aw/25°C) and two suboptimal: 0.90aw/25°C and 0.98aw/37°C as suboptimal water activity and temperature, respectively, which might take place through over ripening and dehydration of grapes. For each condition, 12 Petri dishes were inoculated, and colony growth and OTA production were measured over time. ANOVA revealed significant differences among μ and λ within the 30 assayed isolates. Coefficients of variation (CV%) revealed a wider dispersion of growth rates at 0.90aw/25°C compared to 0.98aw/25°C, and a more than 4-fold higher CV at 0.98aw/37°C compared to 0.98aw/25°C. However, dispersion of lag phases was similar at 0.98aw/25°C and 0.90aw/25°C and wider at 0.98aw/37°C. There were significant differences (p <0.05) among OTA levels (ng/mm2) for the different conditions, values being lower under marginal conditions, and particularly at 0.98aw/37°C. Coefficients of variation (CV%) revealed a wider dispersion of OTA production at 0.90aw/25°C compared to 0.98aw/25°C, while CV at 0.98aw/37°C was similar to that at 0.98aw/25°C. In order to address the strain variability in growth initiation and prove the well-established notion of reducing OTA in foods by preventing fungal growth, a greater number of strains should be included when developing models for conditions that are suboptimal both for aw for OTA production and temperature levels for growth. [ABSTRACT FROM AUTHOR]

Abstract: Predictive models can be a tool to develop strategies to prevent mould development and consequently mycotoxin production. The aims of this work were to assess the impact of a) high/low levels of inoculum and b) optimal/suboptimal environmental conditions on fungal responses based on both kinetic and probabilistic models. Different levels of spore suspensions of Aspergillus carbonarius and Penicillium expansum were prepared and inoculated centrally with a needlepoint load on malt extract agar (MEA) with 50 replicates. While optimum conditions led to a colony diameter increase which followed Baranyi’s function, suboptimal conditions led to different grow functions. In general, growth rate (μ) and lag phase (λ) were normally distributed. Specifically, the growth rate (μ) showed similar distributions under optimal growth conditions, regardless of the inoculum level, while suboptimal aw and temperature conditions led to higher kurtosis distributions, mainly when the inoculum levels were low. Regarding λ, more skewed distributions were observed, mainly when the inoculum levels were low. Probability models were not much affected by the inoculum size. Lower probabilities of growth were in general predicted under marginal conditions at a given time for both strains. The slopes of the probability curves were smaller under suboptimal growth conditions due to wider distributions. Results showed that a low inoculum level and suboptimal conditions lead to high variability of the estimated growth parameters and growth probability. [ABSTRACT FROM AUTHOR]

Abstract: The aim of the present work was to apply mathematical models for the prediction of growth of aflatoxigenic moulds in powdered Capsicum fruits as a function of its water availability. As prevention of fungal growth effectively conduces to prevention of mycotoxin accumulation, the development of models for prediction of growth of mycotoxigenic fungi becomes a key step in risk management. Two aflatoxigenic A. flavus from chilli powder were grown on 3% chilli powder extract agar at different water activity levels and their growth was evaluated over time in terms of colony sizes and ergosterol accumulation. Both variables were modelled over time, and the resulting parameters (growth rates and lag phases) were modelled as a function of water availability using the Rosso cardinal model. Linear logistic regression was also applied to predict the probability of growth over storage time. Both isolates showed a similar pattern of behaviour, with decreasing growth rates and increasing lag phases with decreasing water activity level. While estimation of optimum a w for growth was consistently around 0.97–0.99, the minimum estimated a w varied from 0.82 to 0.88 depending on the isolate and on the parameters used for predictions. Comparing growth rates obtained for colony size and ergosterol accumulation, a linear relationship between them could be observed. The rate of root square ergosterol/colony diameter/unit of time was 0.25–0.27. Probabilities of growth before 10 days over 90% were estimated at a w 0.91, while the safe period could be extended to more than 20 days (22–29 days) if water activity was decreased to an a w =0.87. Finally, the probability of growth is always under 50% when water availability is under a w 0.85, and almost null for A. flavus UdLTA 3.147. It was concluded that for safe production, storage and transport, chillies and chilli powder must be kept under 31% mc (db) (probability of growth <50%). However, growth is unlikely to occur if chilli is kept at approx. 34% for less than 10 days, or at approx. 33% for less than 20 days. Careful hazard analysis and critical control point (HACCP) techniques during raw material production and the subsequent stages of drying, transportation, elaboration and storage are indispensable. [Copyright &y& Elsevier]

Abstract: The aim of this study was to investigate the distribution of fumonisins (B1 B2, and B3) and total aflatoxins (B1, B2, G1, and G2) in various corn processed fractions. 92 batches of whole corn and derived dry-milled fractions (animal feed flour, flaking grits, corn flour and corn meal) and cooked and roasted cornflakes fractions were industrially obtained. Samples were analyzed for both groups of mycotoxins by enzyme-linked immunosorbent assay (ELISA). Dry milling of corn led to a heterogeneous distribution of the two groups of mycotoxins in the different parts of the grain, with increased levels in fractions processed from outer layers (animal feed flour and corn flour) and decreased levels in fractions processed from inner portions, such as corn meal and flaking grits. Levels of fumonisins in cornflakes were lower than 400 μg/kg, the maximum tolerable limit set by the EU. By contrast, three samples of final product were found to exceed the aflatoxin maximum tolerable limit of 4 μg/kg. Animal feed flour showed concentration factors of 317 and 288% for fumonisins and aflatoxins, respectively. Food traceability system was used by the industrial companies which processed corn into breakfast cereals. Nevertheless, even though the use of food traceability, which is defined as the ability to trace any food, feed, food-producing animal or substance that will be used for consumption through all stages of production, processing and distribution, only initial fumonisin contamination of whole corn and contamination of animal feed flour and corn flour were found to be correlated. [Copyright &y& Elsevier]

Abstract: The aim of this study was to characterize Ochratoxin A (OTA) production by Aspergillus carbonarius under different environmental conditions, and to elucidate the diffusion capacity of OTA throughout synthetic medium. One strain belonging to the species A. carbonarius isolated from vine dried fruit was single-point inoculated onto triplicate synthetic nutrient medium plates at two water activities (0.92 and 0.97) and two temperatures (20 and 30°C). Daily radii were measured and OTA production was tested after 4, 7, 10, 14 and 18 days of incubation at four distances from the centre of colony (1–4cm). OTA production was detected mainly at 0.97a w. Earlier production was detected at 30°C (optimum for growth), whereas maximum OTA concentrations were found at 20°C. OTA production was detected from mycelium that was only a few days old and attained its optimum when mycelium was 4–7 days old at 0.97a w. OTA diffusion was observed at 0.92a w and 20°C. Thus OTA production is discernable in young A. carbonarius mycelium and diffusion of the toxin has been shown to occur in a solid substrate. [Copyright &y& Elsevier]

A multianalyte method, using a MS/MS detector, was applied for a simultaneous determination of 23 mycotoxins in 64 beer products purchased from the supermarket in Lleida, Spain. The samples varied by their origin, brewing technology, alcohol content, etc. The results showed that 20.3% of the tested samples were mycotoxin contaminated overpassing the limit of detection (LOD). None of the alcohol-free samples (17%) were contaminated with mycotoxins. The most frequently occurring toxin was zearalenone (ZEN), being quantified in 65% of the positive samples, with levels ranging from 8.24 to 62.96 μg/L. Regarding the co-occurrence of mycotoxins, three samples were found to contain two or more mycotoxins simultaneously. A deterministic approach was used to evaluate the contribution of beer consumption to daily intake and the proportion of the established tolerable daily intake (TDI) for ZEN and deoxynivalenol (DON) and its metabolite deoxynivalenol-3-glucoside (DON-3-G). [ABSTRACT FROM AUTHOR]

The contamination of food commodities by fungal toxins has attracted great interest because many of these mycotoxins are responsible for different diseases, including cancer and other chronic illnesses. Ochratoxin A (OTA) is a mycotoxin naturally present in food, and long-term exposure to food contaminated with low levels of OTA has been associated with renal cancer. In the present study, the cytotoxicity, cytostaticity, and genotoxicity of OTA (0.075–15 µM) in human lymphocytes were evaluated. A comet assay, a modified comet assay (DNA repair assay), which uses N-hydroxyurea (NHU) to detect non-repaired lesions produced by OTA, and a cytokinesis-blocked micronucleus assay were used. Treatments with OTA were not cytotoxic, but OTA caused a cytostatic effect in human lymphocytes at a concentration of 15 µM. OTA (0.075–5 µM) produced a slight increase in the percentage of DNA in the comets and a delay in the DNA repair capacity of the lymphocytes. Micronucleus (MN) induction was observed at OTA concentrations of 1.5 and 5 µM. Our results indicate that OTA induces DNA stable damage at low doses that are neither cytotoxic nor cytostatic, and OTA delays the DNA repair kinetics. These findings indicate that OTA affects two pivotal events in the carcinogenesis pathway. [ABSTRACT FROM AUTHOR]

Abstract: Alternaria is a common contaminating genus of fungi in fruits, grains, and vegetables that causes severe economic losses to farmers and the food industry. Furthermore, it is claimed that Alternaria spp. are able to produce phytotoxic metabolites, and mycotoxins that are unsafe for human and animal health. DNA amplification techniques are being increasingly applied to detect, identify, and quantify mycotoxigenic fungi in foodstuffs, but the inability of these methods to distinguish between viable and nonviable cells might lead to an overestimation of mycotoxin-producing living cells. A promising technique to overcome this problem is the pre-treatment of samples with nucleic acid intercalating dyes, such as propidium monoazide (PMA), prior to quantitative PCR (qPCR). PMA selectively penetrates cells with a damaged membrane inhibiting DNA amplification during qPCRs. In our study, a primer pair (Alt4–Alt5) to specifically amplify and quantify Alternaria spp. by qPCR was designed. Quantification data of qPCR achieved a detection limit of 102 conidia/g of tomato. Here, we have optimized for the first time a DNA amplification-based PMA sample pre-treatment protocol for detecting viable Alternaria spp. cells. Artificially inoculated tomato samples treated with 65μM of PMA, showed a reduction in the signal by almost 7cycles in qPCR between live and heat-killed Alternaria spp. conidia. The tomato matrix had a protective effect on the cells against PMA toxicity, reducing the efficiency to distinguish between viable and nonviable cells. The results reported here indicate that the PMA-qPCR method is a suitable tool for quantifying viable Alternaria cells, which could be useful for estimating potential risks of mycotoxin contamination. [Copyright &y& Elsevier]

Abstract: The mycotoxins aflatoxins (AFs), deoxynivalenol (DON), fumonisins (FBs) and zearalenone (ZEA) are produced by fungal species can contaminate cereal-based food and therefore can be found alone or simultaneously in cereal-based commodities. The objectives of this study were, to study the occurrence of AFs, DON, FBs and ZEA in ethnic and gluten-free food samples from Lleida (Spain), and also to assess the exposure of immigrant and celiac population to the selected mycotoxins. Ethnic food and gluten-free samples were purchased in specialized grocery stores from Lleida (Spain). Mycotoxins were extracted and purified from the samples using immunoaffinity chromatography (IAC) columns. Determination and quantification of AFs, ZEA and FBs was performed with high performance liquid chromatography (HPLC) with fluorescence detector, and coupled to absorbance detector in case of DON. The results showed that FBs were widely found in corn-based foods purchased from special shops specialized on imported foods. DON was mainly found in cuscus and several corn-based ethnic foods. Several gluten-free food samples were contaminated by FBs (6 out of 18) and DON (2 out 18), while AFs and ZEA only were found in 6 ethnic foods samples. Considering these results, these special commodities should not be a health concern for these collectives; however, special attention should be focused on bulk imported foods distributed in special retail shops. [Copyright &y& Elsevier]

Abstract: Cereals are very important for human and animal diet. However, agricultural products can be contaminated by moulds and their mycotoxins. On the other hand, natural plant products with antimicrobial properties could be a possibility to control mycotoxigenic fungi in foods and feeds. In this study, Equisetum arvense and Stevia rebaudiana extracts were tested for their efficacy against a range of mycotoxigenic fungi. Maize agar medium (MAM 2%) was used for this study and E. arvense and S. rebaudiana extracts were added at different concentrations (1–3%) under different water activity (a w) levels (0.85–0.95). Six mycotoxigenic moulds were inoculated and incubated at different temperatures (15–30 °C) during 21 days. In general, no growth was observed with E. arvense extract at 3% in all studied conditions for all isolates. However, with S. rebaudiana extract at 2–3%, growth was not significantly decreased (p < 0.05) in most of the cases. Finally, in terms of toxin production, results were not conclusive. The use of E. arvense extracts as antifungals and antimycotoxigenics and their application should be further explored. [Copyright &y& Elsevier]

Abstract: Patulin, a mycotoxin produced primarily by Penicillium expansum, is currently of great concern because of its undesirable effects in human health. It has been proven that patulin can damage organs and tissues in animals and some studies revealed carcinogenic and teratogenic effects. Patulin is found mainly in low quality apples diverted to production of apple by-products. Apples from cold storage or recently harvested (usually ground harvested or low quality apples) are stored under ambient conditions (deck storage) until they are processed. The present assay studies the consequences of this type of storage in development of lesions and patulin accumulation. The assayed factors were the size of lesions when apples were taken out from cold storage, time the apples stayed at room temperature after cold storage (as a simulation of deck storage) and intraspecific differences between 2 isolates of P. expansum. A sublot of P. expansum inoculated apples was cold stored until lesions achieved concrete sizes. Then, apples were either transferred to a 20 °C storage room for 0 to 5 days or were analysed for patulin immediately. The rest of the apples were directly stored at 20 °C. Each treatment had three replicates. Increase of lesion size with time at 20 °C depended on initial lesion size after cold storage. Bigger lesions were always achieved in apples with bigger initial lesion size. Initial lesion size and time at 20 °C significantly influenced patulin accumulation. No significant amounts of patulin were found in apples with lesions up to 2 cm after cold storage. Patulin amounts significantly increased on the 2nd day at 20 °C day and remained constant until the 5th day. Patulin accumulation tended to be higher when initial lesions were bigger. The assay showed the influence of apple quality (measured as overall lesion size) after cold storage on patulin accumulation during deck storage, as well as the importance of duration of deck storage. Quality of apples entering the processing plant should be assessed in order to program deck storage and minimise patulin accumulation. Predictive models of patulin accumulation as a function of time at room temperature and apple quality should be a useful tool to elaborators. [Copyright &y& Elsevier]

The effect of cinnamon, clove, oregano, palmarosa and lemongrass oils on zearalenone (ZEA) and deoxynivalenol (DON) accumulation by one isolate of Fusarium graminearum in non-sterilized naturally contaminated maize grain at 0.995 and 0.950 aw and at 20°C and 30°C was evaluated at a 500 mg kg−1 level. Efficacy of essential oils was found to be poor, clove essential oil being that with a broader applicability. In general, competing mycoflora seemed to control to a large extent ZEA and DON accumulation, and then the effectiveness of essential oils could only be observed under certain environmental conditions, 0.950 aw/30°C for ZEA, and 0.995 aw/30°C for DON. Better results might be obtained by applying higher dosages; sensory quality, however, should not be compromised. [Copyright &y& Elsevier]

This study aimed to evaluate the mycotoxins transfer in the wort and yeast during ale and lager fermentation, secondary fermentation, and beer storage. Four yeasts were used to ferment wort contaminated with DON and ZEN. The wort was fermented with two S. cerevisiae yeasts (yeasts A and B) at 15, 20, 25 °C for 96 h, and two S. pastorianus (yeasts C and D) at 10, 15, 20 °C for 144 h. After fermentation, the two wort with the highest mycotoxin decrease were selected for a second fermentation and stored period. Mycotoxins were extracted using QuEChERS and analyzed by UHPLC-DAD/FLD. In the wort, DON decreased from 11% to 27%, locating it adsorbed on the yeast from LOD to 6.16%. ZEN in the wort decreased from 29% to 90%, locating it adsorbed on the yeast from 5.41% to 33.86%. Ale-fermentation process obtained the highest decrease of ZEN in the wort, whereas, for DON, there was no significant difference in the reduction between the 2 fermentation styles. Finally, there was no significant change in mycotoxin content during storage. Knowledge of the mitigating effect of fermentation variables could help to reduce the mycotoxin content in beer. • DON and ZEN adsorption ability of S. pastorianus and S. cerevisiae was evaluated. • Temperature and fermentation parameters affect mycotoxins degradation in the brewing. • Second fermentation supplies to the mitigation in a proportion like the first one. • There was no significant change in mycotoxin content during the storage period. [ABSTRACT FROM AUTHOR]