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Plant nutraceuticals have currently been receiving international attention due to their potentially health-promoting effects when consumed as part of a varied diet, including antioxidant, anti-inflammatory, and anti-proliferative properties. These beneficial effects have been attributed to the presence of bioactive compounds naturally occurring in food or produced de novo through metabolic pathways. However, taken into account that nutraceuticals could be concentrated forms of the food or plants, it is probable to find undesirable substances utilized in crop protection, such as pesticides, and other contaminants (e.g. mycotoxins), which could be concentrated during the extraction process. Hence, this review will deepen into the bioactive compounds and health-promoting properties of plant nutraceuticals as well as the challenges related to safety issues because of the increase consumption of these products by population.

The presence of mycotoxins in cereals and cereal products remains a significant issue. The use of natural ingredients such as pumpkin and whey, which contain bioactive compounds, could be a strategy to reduce the use of conventional chemical preservatives. The aim of the present work was to study the bioaccessibility of aflatoxin B1 (AFB1) and ochratoxin (OTA) in bread, as well as to evaluate the effect of milk whey (with and without lactic acid bacteria fermentation) and pumpkin on reducing mycotoxins bioaccessibility. Different bread typologies were prepared and subjected to an in vitro digestion model. Gastric and intestinal extracts were analyzed by HPLC–MS/qTOF and mycotoxins bioaccessibility was calculated. All the tested ingredients but one significantly reduced mycotoxin intestinal bioaccessibility. Pumpkin powder demonstrated to be the most effective ingredient showing significant reductions of AFB1 and OTA bioaccessibility up to 74% and 34%, respectively. Whey, fermented whey, and the combination of pumpkin-fermented whey showed intestinal bioaccessibility reductions between 57–68% for AFB1, and between 11–20% for OTA. These results pointed to pumpkin and milk whey as potential bioactive ingredients that may have promising applications in the bakery industry.

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

Mycotoxins are secondary fungal metabolites that can have toxic effects on both human and animals, mainly due to their exposure through food. Presence of these compounds has been demonstrated in a wide variety of raw materials, foods and feeds, in which it is common to find co-occurrence of different mycotoxins, in small amounts, which can generate subchronic toxic effects, as well as bioaccumulation. This article reviews the main elements that shape the problem of mycotoxins for human and animals, and explains the future challenges that will arise in the study of mycotoxins, among which are the effects that climate change can have on the pattern of contamination by mycotoxins in food, the growing discovery of new modified mycotoxins, the evaluation of the co-occurrence of these toxins and other chemical contaminants, and the ways to detect and eliminate these toxic compounds from food., Las micotoxinas son metabolitos fúngicos secunda­rios que pueden ejercer un efecto tóxico tanto en el hombre como en los animales debido, principalmente, a su exposición a través de los alimentos. La presencia de estos compuestos ha sido demostrada en una amplia variedad de materias primas, alimentos y piensos, en los que lo habitual es encontrar de for­ma frecuente una contaminación múltiple por diferentes mico­toxinas, en pequeñas cantidades, lo que puede generar efectos tóxicos subcrónicos, así como bioacumulación. Este artículo revisa los principales elementos que configuran la problemática de las micotoxinas para el hombre y los animales, y aborda los retos de futuro que se plantean en el estudio de las micotoxinas, entre los que destacan el efecto que el cambio climático puede tener sobre el patrón de contaminación por micotoxinas en los alimentos, el descubrimiento creciente de nuevas micotoxinas en formas modificadas, la evaluación de la coexistencia de estas to­xinas y otros contaminantes, y las formas para detectar e intentar eliminar estos compuestos tóxicos de los alimentos.

Due to the globalization, mycotoxins have been considered a major risk to human health being the main con- taminants of foodstuffs. Among them, AFB1 and OTA are the most toxic and studied. Therefore, the goal of this review is to deepen the knowledge about the toxicological effects that AFB1 and OTA can induce on human health by using flow cytometry and immunofluorescence techniques in vitro and in vivo models. The examination of the selected reports shows that the majority of them are focused on immunotoxicity while the rest are con- cerned about nephrotoxicity, hepatotoxicity, gastrointestinal toxicity, neurotoxicity, embryotoxicity, reproduc- tive system, breast, esophageal and lung toxicity. In relation to immunofluorescence analysis, biological processes related to AFB1- and OTA-toxicity were evaluated such as inflammation, neuronal differentiation, DNA damage, oxidative stress and cell death. In flow cytometry analysis, a wide range of assays have been performed across the reviewed studies being apoptosis assay, cell cycle analysis and intracellular ROS measurement the most employed. Although, the toxic effects of AFB1 and OTA have been reported, further research is needed to clarify AFB1 and OTA-mechanism of action on human health.

The world requests for raw materials used in animal feed has been steadily rising in the last years driven by higher demands for livestock production. Mycotoxins are frequent toxic metabolites present in these raw materials. The exposure of farm animals to mycotoxins could result in undesirable residues in animal-derived food products. Thus, the potential ingestion of edible animal products (milk, meat and fish) contaminated with mycotoxins constitutes a public health concern, since they enter the food chain and may cause adverse effects upon human health. The present review summarizes the state-of-the-art on the occurrence of mycotoxins in feed, their metabolism and carry-over into animal source foodstuffs, focusing particularly on the last decade. Maximum levels (MLs) for various mycotoxins have been established for a number of raw feed materials and animal food products. Such values are sometimes exceeded, however. Aflatoxins (AFs), fumonisins (FBs), ochratoxin A (OTA), trichothecenes (TCs) and zearalenone (ZEN) are the most prevalent mycotoxins in animal feed, with aflatoxin M1 (AFM1) predominating in milk and dairy products, and OTA in meat by-products. The co-occurrence of mycotoxins in feed raw materials tends to be the rule rather than the exception, and the carry-over of mycotoxins from feed to animal source foods is more than proven.

Food bio-preservatives are requested as substituents of chemical pesticides in food. The aim of this study was to carry out a screening of twenty biocontrol agents (BCAs) for their potential fungicidal activity in vitro. Twenty BCAs were tested against ten pathogenic fungi. Some of the cell-free supernatants (CFS) tested showed in vitro antifungal activity versus pathogenic fungi. The highest fungicidal activity was observed in the fermented CFS of Paenibacillus chibensis CECT 375, Bacillus amyloliquefaciens CECT 493, and Pantoea agglomerans CECT 850, which showed a minimum inhibitory concentration (MIC) and minimum fungicidal concentration (MFC) values of 125 and 250 g/L, respectively. The compounds responsible for the antifungal activity, such as organic and phenolic acids, were determined. Lactic acid, acetic acid, benzoic acid, and phenyllactic acid among others can be related to antifungal activity. HPLC-MS/MS analysis showed a reduction of ochratoxin A (OTA) and aflatoxin B1 (AFB1) up to 26% (Paenibacillus alvei CECT 2) and 55% (Paenibacillus polymyxa CECT 155), respectively. The present study prompts that metabolism products of BCAs are propitious for the bioconservation of food, due to their ability to reduce the proliferation of mycotoxigenic fungi and mycotoxins production.

Review Contamination of animal feed with mycotoxins still occurs very often, despite great efforts in preventing it. Animal feeds are contaminated, at low levels, with several mycotoxins, particularly with those produced by Aspergillus and Fusarium genera (Aflatoxin B1, Ochratoxin A, Zearalenone, Deoxynivalenol and Fumonisina B1). In animal feed, to date, only Aflatoxin B1 is limited through EU regulation. Consequently, mycotoxins cause serious disorders and diseases in farm animals. In 2009, the European Union (386/2009/EC) approved the use of mycotoxin-detoxifying agents, as feed additives, to prevent mycotoxicoses in farm animals. The present review gives an overview of the problem of multi-mycotoxin contamination of feed, and aims to classify mycotoxin adsorbing agents (minerals, organic, and synthetic) for feed decontamination, focusing on adsorbents with the ability to bind to multiple mycotoxins, which should have a more effective application in farms but they are still little studied in scientific literature. The authors are grateful to the Spanish government (projects RTC-20153508-2 and AGL2014-52648-REDT) and to XaRTA (The Catalonian Reference Network in Food Technology) for funding. They also would like to thank Arthur Kendall for editing the English usage in the manuscript. P. Vila-Donat thanks MINECO and the University of Lleida for her postdoctoral contract.

Alternariol (AOH) is a mycotoxin produced by Alternaria spp. Soyasaponin I (Ss-I) is present naturally in legumes, and it has antioxidant properties. Cytotoxic and genotoxic effects of AOH have been demonstrated previously in vitro. In the present study, the cytotoxicity of AOH, Ss-I, and soyasaponins-rich extract from lentils was investigated; as well as, the cytoprotective effects of Ss-I and lentil extracts against AOH induced-cytotoxicity on Caco-2 cells. Cytotoxicity was carried out using MTT and PC assays (AOH: 3.125–100 µM, Ss-I: 3.125–50 µM, and lentil extracts: 1:0–1:32) during 24 h of exposure. Only AOH showed cytotoxic effect. The reduction in cell proliferation ranged from 25% to 47%. Simultaneous combination of Ss-I with AOH (1:1) increased cell proliferation (35%) compared to AOH tested alone. The Ss-I and extracts showed synergistic cytoprotective effects against cytotoxicity induced by AOH on Caco-2 cells. Food commodities containing Ss-I could contribute to diminish the toxicological risk that natural contaminant as AOH in diet can produce to humans.

cope : the aim of our work was to produce a hydroalcoholic extract of lentils and to examine: a) the hypocholesterolemic action in an animal model, by studying the plasma cholesterol level and the concentration of bile acids in the faeces; b) the potential prebiotic effect, by conducting an in vitro culture fermentation experiment and assessing the level of short chain fatty acids (SCFAs) in the faeces of rats. Methods and results : Lentil extract (LE) was obtained by extracting lentils with a solution of H20/EtOH (70/30 v/v) for 3 h, and the content of main nutrients was determined. After 71 days of diet-induced hypercholesterolemia in rats, LE reduced the cholesterol level of rats of 16.8% (p

In the present work different clays have been characterized according to their mycotoxins adsorbent ability. Firstly, 27 bentonite clays from different geographical origins were evaluated, at 0.02% w/v, using an in vitro screening method versus five mycotoxins (aflatoxin B1, AFB1; deoxynivalenol, DON; ochratoxin A, OTA; fumonisin B1, FB1; and, zearalenone, ZEN) by Enzyme- Linked Immunosorbent Assay (ELISA). Subsequently, 7 bentonite clays (6 of which were trioctahedral bentonites) selected from the preliminary test, and 7 commercial adsorbent products were subjected to an in vitro equilibrium adsorption experiment (at 0.02% w/v) against six concentrations of AFB1 (0.02-4 mg/L), and OTA (0.05-1 mg/L) by using simulated gastrointestinal (GI) juices, and successively analysed by HPLC-FD. Equilibrium isotherm functions were fitted to the data by nonlinear regression analysis. In vitro adsorption equilibrium experiments showed that AFB1 adsorption was very high with all the adsorbents tested. In particular, the seven pre-selected bentonites adsorbed most of the AFB1 present at the lower level tested, while only three of these reached more than 50% of OTA adsorption. Adsorption increased inversely to the toxin concentration and both Langmuir and Freundlich isotherm models fitted well to the data. Generally, the pre-selected bentonites (B1-B7) showed better mycotoxin adsorption than commercial products (C1-C7) at all levels of mycotoxins tested. The 10-fold dose increase of the best tri-octahedral bentonite (B4) rendered a more effective adsorption of OTA, reaching almost 75% of adsorption (at pH 5) The authors are grateful to the following institutions: Spanish government (projects AGL2014-52648-REDT and RTC-2015-3508-2–ERDF. A way to make Europe, funded by the Ministry of Economy and Competitiveness of the Government of Spain and by the European Regional Development Fund of the European Union with the objective of “Promoting technological development, innovation and quality research”) and to XaRTA (The Catalonian Reference Network in Food Technology) for funding. P. Vila-Donat thanks MINECO for her postdoctoral contract.

This study sought to contribute to the assessment of the nutritional properties of legumes by determining the fatty acid (FA) composition of 29 legume samples after the evaluation of nine extraction methods. The Folch method and liquid-solid extraction with hexane/isopropanol or with hexane/acetone were investigated, as was the effect of previous hydration of samples. Soxhlet extractions were also evaluated with different solvent mixtures. Results on FA composition using the hexane/isopropanol extraction method were the same in terms of FA composition of the Folch method, but the extraction yield was only around 20-40% of that of the Folch method preceded by hydration. Some types of legumes showed particularly interesting values for the ratio of polyunsaturated fatty acids (PUFAs) n-6/n-3, such as lentils, with the value of 4.0, and Azuki beans, at 3.2. In lentils, the PUFAs% ranged from 42.0% to 57.4%, while in Azuki beans it was 57.5%.

Legumes are an excellent source of macronutrients and phytochemicals as isoflavones. The aim of this work was to develop a new analytical method for determining five isoflavone compounds, three of which are aglycons, namely daidzein, genistein, biochanin A, and two of which, daidzin and genistin, are glycosilated, in lentils and other pulses, using an effective clean-up system and UHPLC-MS/MS (triple quadrupole) method. The recoveries obtained by spiking the lentil samples with a standard mixture of isoflavones at three levels of fortification (5, 25 and 100 μg kg(-1)) were in the range of 54.4-111.1%, 68.6-91.1%, and 84.4-114%, respectively. The method was applied to analyse 48 lentil samples from central Italy and pulses for determining the isoflavone content, which was found to range from 1.1 to 95.6 μg kg(-1).

In this work, an innovative and fast analytical method for the quantification of soyasaponins I and βg in lentils has been developed. Samples were extracted using 70 % aqueous ethanol at room temperature and then injected into a high-performance liquid chromatography–tandem mass spectrometry system. The correlation coefficients of calibration curves of the analyzed compounds were ≥0.9997. The recoveries obtained by spiking the lentil samples with a standard mixture of soyasaponins I and βg at 50 and 100 mg l−1 were in the range of 96–101 and 98–103 %, respectively. The validated method was applied to the analysis of 30 lentil samples from central Italy. Soyasaponins I and βg were present in these lentils in concentrations that ranged from 54 to 226 mg kg−1 and from 436 to 1,272 mg kg−1, respectively. Our data indicated that lentils cultivated in fields at intermediate altitudes (1,142–1,387 m) showed the highest levels of soyasaponins, a finding confirmed by principal component analysis.

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 B1 (FB1: 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 FB1 (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 FB1, reaching >90% of adsorption. The present work proposes the use of some tri-octahedral bentonites as feed additives for Fusarium-mycotoxin reduction. The authors are grateful to the following institutions: Spanish government (projects AGL2014-52648-REDT and RTC-2015-3508-2–ERDF. A way to make Europe, funded by the Ministry of Economy and Competitiveness of the Government of Spain and by the European Regional Development Fund of the European Union with the objective of “Promoting technological development, innovation and quality research”) and to XaRTA (The Catalonian Reference Network in Food Technology) for funding. P. Vila-Donat thanks MINECO for her postdoctoral contract.

Corn (Zea mays) is a worldwide crop subjected to infection by toxigenic fungi such as Fusarium verticillioides during the pre-harvest stage. Fusarium contamination can lead to the synthesis of highly toxic mycotoxins, such as Fumonisin B1 (FB1) and Fumonisin B2 (FB2), which compromises human and animal health. The work aimed to study the antifungal properties of fermented yellow and oriental mustard extracts using nine lactic acid bacteria (LAB) in vitro. Moreover, a chemical characterization of the main phenolic compounds and organic acids were carried out in the extracts. The results highlighted that the yellow mustard, fermented by Lactiplantibacillus plantarum strains, avoided the growth of Fusarium spp. in vitro, showing Minimum Inhibitory Concentration (MIC) and Minimum Fungicidal Concentration (MFC) values, ranging from 7.8 to 15.6 g/L and 15.6 to 31.3 g/L, respectively. Then, the lyophilized yellow mustard fermented extract by L. plantarum TR71 was applied through spray-on corn ears contaminated with F. verticillioides to study the antimycotoxigenic activity. After 14 days of incubation, the control contained 14.71 mg/kg of FB1, while the treatment reduced the content to 1.09 mg/kg (92.6% reduction). Moreover, no FB2 was observed in the treated samples. The chemical characterization showed that lactic acid, 3-phenyllactic acid, and benzoic acid were the antifungal metabolites quantified in higher concentrations in the yellow mustard fermented extract with L. plantarum TR71. The results obtained confirmed the potential application of fermented mustard extracts as a solution to reduce the incidence of mycotoxins in corn ears.