Development of a liquid chromatography–tandem mass spectrometry for the determination of enniatins in biological samples from Wistar rats

FoodSimplex as a Mean to Improve Portuguese Restaurants’ Goods Manufacturing Practices - Audit and Microbial Assessment

Background: The consumption of meals at resturants has increased mass catering establishments. Such food services may present several risk factors for foodborne diseases, which have a substantial negative impact on public health. The smallest business with no technical or financial support struggles to comply with food safety European Regulation. The aim of this four years study was to assess Good Manufacture Practices (GMP) in Portuguese restaurants applying a designed food safety methodology, FoodSimplex, through audit data to determine good manufacture practices, and microbiological sampling of the meals before and after its implementation, by food groups according to their risk. Method…

Mycotoxin Analysis of Human Urine by LC-MS/MS: A Comparative Extraction Study

The lower mycotoxin levels detected in urine make the development of sensitive and accurate analytical methods essential. Three extraction methods, namely salting-out liquid–liquid extraction (SALLE), miniQuEChERS (quick, easy, cheap, effective, rugged, and safe), and dispersive liquid–liquid microextraction (DLLME), were evaluated and compared based on analytical parameters for the quantitative LC-MS/MS measurement of 11 mycotoxins (AFB1, AFB2, AFG1, AFG2, OTA, ZEA, BEA, EN A, EN B, EN A1 and EN B1) in human urine. DLLME was selected as the most appropriate methodology, as it produced better validation results for recovery (79–113%), reproducibility (RSDs < 12%), and repeatability (RSDs…

HPLC-UV/Vis-APCI-MS/MS Determination of Major Carotenoids and Their Bioaccessibility from "Delica" (Cucurbita maxina) and "Violina" (Cucurbita moschata) Pumpkins as Food Traceability Markers

Carotenoids are a widespread group of fat-soluble pigments, and their major nutritional importance comes from their pro-vitamin A activity and their antioxidant capacity. In this study, two different pumpkin cultivars (Cucurbita maxima, also named `Delica&rsquo

Toxicity of mycotoxins in vivo on vertebrate organisms: A review.

Mycotoxins are considered to be a major risk factor affecting human and animal health as they are one of the most dangerous contaminants of food and feed. This review aims to compile the research developed up to date on the toxicological effects that mycotoxins can induce on human health, through the examination of a selected number of studies in vivo. AFB1 shows to be currently the most studied mycotoxin in vivo, followed by DON, ZEA and OTA. Scarce data was found for FBs, PAT, CIT, AOH and Fusarium emerging mycotoxins. The majority of them concerned the investigation of immunotoxicity, whereas the rest consisted in the study of genotoxicity, oxidative stress, hepatotoxicity, cytotoxicity,…

Evolution of emerging Fusarium mycotoxins contents throughout the shelf-life period of food

which is cooked by steaming. It has been historically eaten in North African countries but nowadays its consumption is widely extended all around the world. As a cereal-based food, semolina is highly susceptible to contamination by mycotoxin-producing fungi. The presented procedure involves a modified QuEChERSbased extraction of 24 mycotoxins (15-ADON, 3-ADON, AFLAB1, AFLAB2, AFLAG1, AFLAG2, BEA, DON, DAS, ENA, ENA1, ENB, ENB1, FB1, FB2, FB3, FUSX, HT-2, NEO, NIV, OTA, STG, T2, ZEA) produced by Aspergillus, Penicillum and Fusarium fungi. The validation was performed by analyzing recovery samples at three different spiked concentrations with four replicates (n=4) at each concentration. Recov…

Bioaccessibility and bioavailability of fumonisin B2 and its reaction products with isothiocyanates through a simulated gastrointestinal digestion system

Abstract Fumonisins (FBs) are toxins produced mainly by the molds Fusarium verticillioides (also known as Fusarium moniliforme) and Fusarium proliferatum. These mycotoxins are contaminants of wheat, maize, maize-based foods and other grains worldwide. Isothiocyanates (ITCs) are natural compounds produced by the enzymatic hydrolysis of glucosinolates, which are found in plants of the Brassicaceae family. The use of ITCs as food preservatives has been extensively researched. In this study, allyl (AITC), phenyl (PITC) and benzyl isothiocyanates (BITC) fumigation systems (500 μL/L) were employed to reduce FB2 levels naturally produced in bread by Gibberella moniliformis CECT 2987. Reaction prod…

Procedimento estratégico em três estágios de seleção de variáveis para a obtenção de resultados equilibrados na pesquisa em saúde pública

Multidisciplinary research in public health is approached using methods from many scientific disciplines. One of the main characteristics of this type of research is dealing with large data sets. Classic statistical variable selection methods, known as “screen and clean”, and used in a single-step, select the variables with greater explanatory weight in the model. These methods, commonly used in public health research, may induce masking and multicollinearity, excluding relevant variables for the experts in each discipline and skewing the result. Some specific techniques are used to solve this problem, such as penalized regressions and Bayesian statistics, they offer more balanced results a…

Mycotoxins and their consequences in aquaculture: A review

Fish consumption has been increasing worldwide, mainly due to the availability, access and price in relation to other kinds of meat consumption, such as beef, pork, and poultry. Consequently, some concerns begin to emerge, primarily regarding the quality of fish available in the market. Residues could be present in any product of animal origin causing economic losses and putting into a risk human and animal health. Food contamination by mycotoxins is a risk to human and animal health, and it is responsible for significant economic losses. It's very difficult to prove that a disease is a mycotoxicosis, and even when mycotoxins are detected, it is not easy to show that they are the etiologica…

Analysis of trichothecenes in laboratory rat feed by gas chromatography-tandem mass spectrometry

A method for the determination of seven trichothecenes, neosolaniol (NEO), diacetoxyscirpenol (DAS), deoxynivalenol (DON), nivalenol (NIV), fusarenon-X (FUS-X), 3-acetyldeoxynivalenol (3-ADON) and 15-acetyldeoxynivalenol (15-ADON), in laboratory rat feed by GC-MS/MS was developed. Sample extraction and purification was performed by an acidified mixture of acetonitrile/water (80-20% v/v). Limits of quantitation (LOQs) were between 1 and 10 μg kg(-1) for all studied trichothecenes. Eight concentration levels between the LOQ and 100 × LOQ were used for the calibration curves. Matrix-matched calibration was used for quantitation purposes to compensate the detector signal enhancement obtained fo…

Transcriptional Changes after Enniatins A, A1, B and B1 Ingestion in Rat Stomach, Liver, Kidney and Lower Intestine

Enniatins (ENs) are depsipeptide mycotoxins produced by Fusarium fungi. They are known for their capacity to modulate cell membrane permeability and disruption of ionic gradients, affecting cell homeostasis and initiating oxidative stress mechanisms. The effect of the acute toxicity of ENs A, A1, B and B1 at two different concentrations after 8 h of exposure was analysed in Wistar rats by a transcriptional approach. The following key mitochondrial and nuclear codified genes related to the electron transport chain were considered for gene expression analysis in stomach, liver, kidney and lower intestine by quantitative Real-Time PCR: mitochondrially encoded NADH dehydrogenase 1 (MT-ND1), mit…

Amylase–Trypsin Inhibitors in Wheat and Other Cereals as Potential Activators of the Effects of Nonceliac Gluten Sensitivity

Nonceliac gluten sensitivity (NCGS) is a gluten-related gastrointestinal disorder distinct from celiac disease (CD) and gluten allergy that is not easy to diagnose due to the lack of biomarkers. It is characterized by intestinal symptoms and extraintestinal manifestations with the consumption of gluten-containing foods. In contrast to CD, NCGS patients do not present a genetic predisposition or intestinal villi atrophy. Recent studies question the proinflammatory triggering activity of α-gliadin fraction contained in wheat, since it has been demonstrated that the amylase-trypsin inhibitors (ATIs) exert a strong activating effect on the innate immune response. We aimed to analyze the role of…

Influence of prebiotics, probiotics and protein ingredients on mycotoxin bioaccessibility

The aim of this study was to investigate the influence of prebiotic compounds (cellulose and inulin), food ingredients (milk whey, β-lactoglobulin and calcium caseinate) and several probiotic microorganisms on the bioaccessibility of beauvericin (BEA), enniatins (ENs A, A1, B, B1), deoxynivalenol (DON) and zearalenone (ZEA) present in wheat crispy bread produced with wheat flour previously fermented with F. tricinctum, F. culmorum and G. zeae. The bioaccessibility of mycotoxins was determined by a dynamic simulated gastrointestinal digestion system, imitating the human digestive physiological conditions of the gastrointestinal tract. Mycotoxins were determined in the simulated intestinal fl…

Transcriptional study after Beauvericin and Enniatin B combined exposure in Jurkat T cells

Simultaneous mycotoxins toxicity is complex and non-predictable based on their individual toxicities. Beauvericin and Enniatins are emerging mycotoxins highly co-occurrent in food and feed, and their cytotoxicity has been reported in several human cell lines. RNA-seq studies of individual exposure in Jurkat cells demonstrated human genome perturbation mainly affecting mitochondrial pathways, however, both mycotoxins showed differences between their toxic responses. This study investigates the transcriptional effects of combined exposure to Beauvericin and Enniatin B (1:1) (0.1, 0.5, 1.5 μM; 24 h) in Jurkat cells by qPCR on 30 selected target genes (10 mitochondrial, 20 nuclear). Gene expres…

Transcriptomic study of the toxic mechanism triggered by beauvericin in Jurkat cells

Beauvericin (BEA), an ionophoric cyclic hexadepsipeptide mycotoxin, is able to increase oxidative stress by altering membrane ion permeability and uncoupling oxidative phosphorylation. A toxicogenomic study was performed to investigate gene expression changes triggered by BEA exposure (1.5, 3 and 5 mu M; 24 h) in Jurkat cells through RNA-sequencing and differential gene expression analysis. Perturbed gene expression was observed in a concentration dependent manner, with 43 differentially expressed genes (DEGs) overlapped in the three studied concentrations. Gene ontology (GO) analysis showed several biological processes related to electron transport chain, oxidative phosphorylation, and cel…

A preliminary study in Wistar rats with enniatin : A contaminated feed

A 28-day repeated dose preliminary assay, using enniatin A naturally contaminated feed through microbial fermentation by a Fusarium tricinctum strain, was carried out employing two months-old female Wistar rats as in vivo experimental model. In order to simulate a physiological test of a toxic compound naturally produced by fungi, five treated animals were fed during twenty-eight days with fermented feed. As control group, five rats were fed with standard feed. At the 28th day, blood samples were collected for biochemical analysis and the gastrointestinal tract, liver and kidneys were removed from each rat for enniatin A detection and quantitation. Digesta were collected from stomach, duode…

Transcriptional profiling reveals functional links between RasGrf1 and Pttg1 in pancreatic beta cells

This article is published under license to BioMed Central Ltd. This is an Open Access article distributed under the terms of the Creative Commons Attribution License .

Influence of pro- and prebiotics on gastric, duodenal and colonic bioaccessibility of the mycotoxin beauvericin

Abstract Beauvericin (BEA) is a bioactive compound produced by the secondary metabolism of several Fusarium strains and known to have various biological activities. This study investigates the influence of several dietary fibers (galactomanan, glucomannan, citrus fiber, bamboo fiber, carrot fiber, pie fiber, β-glucan, xilan, and cellulose) and probiotic strains ( Lactobacillus animalis , Lb. casei , Lb. casei , Lb. plantarum , Lb. rhuminis , Lb. casei casei , Bifidobacterium breve , Bf. Adolescents , Bf. bifidum , Corynebacterium vitaeruminis , Streptococcus faecalis , Eubacterium crispatus , and Saccharomyces cerevisiae ) on the minor Fusarium mycotoxin BEA bioaccessibility employing a mod…

A chemical approach for the reduction of beauvericin in a solution model and in food systems.

Abstract Beauvericin (BEA) is a bioactive compound produced by the secondary metabolism of several Fusarium strains with a strong antibacterial, antifungal, and insecticidal activities. This study evaluated the reduction of BEA added at 25 mg/kg in phosphate buffer saline (PBS) solutions at pH of 4, 7 and 10, or to different cereal products (kernels and flours) by the bioactive compounds phenyl isothiocyanate (PITC) and benzyl isothiocyanate (BITC). The concentration of the mycotoxin was evaluated using liquid chromatography coupled to the diode array detector (LC-DAD). In solution, BEA reduction ranged from 9% to 94% on a time-dependent fashion and lower pH levels resulted in higher BEA re…

Spatial learning and long-term memory impairments in RasGrf1 KO, Pttg1 KO, and double KO mice

© 2018 The Authors.

In vitro antifungal activity of allyl isothiocyanate (AITC) against Aspergillus parasiticus and Penicillium expansum and evaluation of the AITC estimated daily intake

Abstract Isothiocyanates (ITCs) are natural compounds derived from cruciferous vegetables produced by enzymatic conversion of metabolites called glucosinolates. They are potentially useful antimicrobial compounds for food applications have been shown to be promising agents against cancer in human cell culture, animal models, and in epidemiological studies. In this study, the antifungal activity of the allyl isothiocyanate (AITC) was evaluated on two mycotoxigenic fungi as Aspergillus parasiticus and Penicillium expnsum , aflatoxins (AFs) and patulin (PAT) producers, employing an assay on solid medium. Also an approximation of the risk evaluation associated to the intake of food treated with…

Reduction of the aflatoxins B1, B2, G1 and G2 in Italian piadina by isothiocyanates

Abstract Aflatoxins (AFs) are mycotoxins produced mainly by the molds Aspergillus flavus , Aspergillus parasiticus and A . nomius . These mycotoxins are contaminants of cereals. AFB 1 , the most abundant and toxic metabolite, is known to cause several toxic responses, such as hepatotoxicity, teratogenicity and mutagenicity. Isothiocyanates (ITCs) are natural compounds produced by the enzymatic hydrolysis of glucosinolates (GLs), which have shown potent antimicrobial activity in food applications. In this study, ITCs derived from oriental and yellow mustard (0.1, 0.5 and 1 g of flour) were used to avoid the production of AFs in piadina (a typical Italian flatbread) contaminated with A . para…

Estudios clínicos sobre la enfermedad celíaca (2014-2019): revisión sistemática de la prevalencia de la presentación clínica y enfermedades asociadas por edades

Introducción: La enfermedad celíaca (EC) se caracteriza por una gran variedad de signos, síntomas y enfermedades asociadas en su forma de presentación, incluso puede cursar de forma asintomática. Recientes estudios muestran la variación del espectro clínico según la edad. En niños y niñas pequeñas predomina la forma clásica con síntomas como distensión abdominal, disminución del apetito, diarrea y pérdida de peso. Las manifestaciones atípicas frecuentes en niños mayores son dolor abdominal, estreñimiento, reflujo, vómitos, fatiga, talla baja. En adultos se observa una reducción de la forma de presentación clásica e incremento de la no clásica.Material y métodos: Se realizó una revisión sist…

Multi-mycotoxin analysis in North African semolina samples using a modified QuEChERS-based extraction procedure and HPLC–MS/MS

Multi-mycotoxin contamination of couscous semolina commercialized in Morocco.

The multi-mycotoxin contamination of ninety-eight (98) couscous semolina samples collected from various areas in Morocco was investigated in this study. Samples were surveyed for the presence of 22 mycotoxins (four aflatoxins, ochratoxin A, diacetoxiscyrpenol (DAS), three fumonisins, beauvericin (BEA), deoxynivalenol (DON), 15-acetyl-deoxynivalenol (15-ADON), 3-acetyl-deoxynivalenol (3-ADON), nivalenol (NIV), sterigmatocystin (STG), zearalenone (ZEA), four enniatins, T-2 and HT-2 toxins). Results showed that 96 out of 98 total couscous samples (98%) were contaminated by at least one mycotoxin. Enniatin B (ENB), Enniatin B1 (ENB1), Enniatin A1 (ENA1) and zearalenone (ZEA) have shown the high…

Mitochondrial proteomics profile points oxidative phosphorylation as main target for beauvericin and enniatin B mixture

Abstract Beauvericin (BEA) and enniatin B (EN B) are non-legislated Fusarium mycotoxins usually found in cereal and cereal-based products all around the world. By the proteomic analysis of mitochondria enriched extracts from Jurkat cells exposed for 24 h to three concentrations of BEA:EN B (0.01–0.1–0.5 μM), a number of 1821 proteins (202 mitochondrial) were identified and relatively quantified. 340 proteins (59 mitochondrial) were statistically significant altered in our samples (Anova p-value ≤ 0.05 and fold change (FC) ≥1.5). The protein mitochondrial translational release factor 1 like (MTRF1L) was the most abundant protein in the three mycotoxin exposures studied. The mycotoxins mixtur…

Occurrence of mycotoxins in laboratory rat feeds

Comparative study of two sample treatments for a liquid chromatography–tandem mass spectrometry determination of mycotoxin biomarkers in urine

Antifungal activity of gaseous allyl, benzyl and phenyl isothiocyanate in vitro and their use for fumonisins reduction in bread

Abstract Fumonisins (FBs) are toxins produced mainly by the molds Fusarium verticillioides (synonym Fusarium moniliforme ) and Fusarium proliferatum . These mycotoxins are contaminants of wheat, maize, maize-based foods and other grains worldwide. FB 1 , the most abundant and toxic metabolite, is known to cause a range of species-specific toxic responses. It has been associated with a high incidence of esophageal cancer in the Transkei region of South Africa and some provinces of China. In addition, FB 1 has been declared a class 2B carcinogen or “possibly carcinogenic to humans”. Isothiocyanates (ITCs) are natural compounds produced by enzymatic conversion of metabolites called glucosinola…

Determination of melatonin in Acyrthosiphon pisum aphids by liquid chromatography-tandem mass spectrometry.

Melatonin is a hormone mainly involved in the regulation of circadian and seasonal rhythms in both invertebrates and vertebrates. Despite the identification of melatonin in many insects, its involvement in the insect seasonal response remains unclear. A liquid chromatography tandem mass spectrometry (LC-MS/MS) method has been developed for melatonin analysis in aphids (Acyrthosiphon pisum) for the first time. After comparing two different procedures and five extraction solvents, a sample preparation procedure with a mixture of methanol/water (50:50) was selected for melatonin extraction. The method was validated by analyzing melatonin recovery at three spiked concentrations (5, 50 and 100 p…

Blood, breast milk and urine: potential biomarkers of exposure and estimated daily intake of ochratoxin A: a review.

The purposes of this review are to study potential biomarkers of exposure for ochratoxin A (OTA) in biological fluids (blood, urine and breast milk) for the period 2005-14, calculate the estimated daily intake (EDI) of OTA by using database consumption for the Spanish population, and, finally, to correlate OTA levels detected in blood and EDI values calculated from food products. The values of OTA detected in potential biomarkers of exposure for blood, breast milk and urine ranged from 0.15 to 18.0, from 0.002 to 13.1, and from 0.013 to 0.2 ng ml(-1), respectively. The calculated EDI for OTA in plasma ranged from 0.15 to 26 ng kg(-1) bw day(-1), higher than that obtained in urine (0.017-0.4…

In vitro and in vivo evaluation of AFB1 and OTA-toxicity through immunofluorescence and flow cytometry techniques: A systematic review

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 toxi…

Enniatin B induces expression changes in the electron transport chain pathway related genes in lymphoblastic T-cell line

Abstract Enniatin B is a ionophoric and lipophilic mycotoxin which reaches the bloodstream and has the ability to penetrate into cellular membranes. The purpose of this study was to reveal changes in the gene expression profile caused by enniatin B in human Jurkat lymphoblastic T-cells after 24 h of exposure at 1.5, 3 and 5 μM by next generation sequencing. It was found that up to 27% of human genome expression levels were significantly altered (5750 genes for both down-regulation and up-regulation). In the three enniatin B concentrations studied 245 differentially expressed genes were found to be overlapped, 83 were down and 162 up-regulated. ConsensusPathDB analysis of over-representation…

Bioaccessibility and bioavailability of bioactive compounds from yellow mustard flour and milk whey fermented with lactic acid bacteria.

Microbial fermentation with lactic acid bacteria (LAB) is a natural food biopreservation method. Yellow mustard and milk whey are optimum substrates for LAB fermentation. The aim of the present study was to evaluate the bioaccessibility and bioavailability of bioactive compounds from yellow mustard flour and milk whey both with and without LAB fermentation. All extracts were subjected to a simulated digestion process. Total polyphenols, DL-3-phenyllactic acid (PLA), lactic acid, and the antioxidant activity were determined in the studied matrices before and after simulated digestion. Yellow mustard flour was significantly richer in total polyphenols, whereas significantly higher concentrati…

In vitro antifungal activity of lactic acid bacteria against mycotoxigenic fungi and their application in loaf bread shelf life improvement

Food spoilage caused by mycotoxigenic fungi represents an important food safety problem. Lactic acid bacteria (LAB) are used as starter cultures in a larger number of food products. In this study, 16 strains of LAB were cultivated in MRS broth under anaerobiosis. Then, cell free supernatants were obtained by centrifugation and their antifungal activity against Aspergillus parasiticus and Penicillium expansum was tested using the disc-diffusion method. Furthermore, the LABs that showed in vitro antifungal activity were used in bread fermentation with yeast in order to study fungal growth inhibition and aflatoxin (AF) reduction in processed bread previously inoculated with A. parasiticus. The…

Mycotoxins in dry-cured meats: A review

Dry-cured meats products are consumed in various regions of the world and, consumers are increasingly demanding better quality and safety of these products. Some fungal species can produce mycotoxins in drycured meats, such as aflatoxins and ochratoxins, which, when ingested, can produce carcinogenic and mutagenic effects in humans. Contamination of these products can occur at different points of the production chain, from the field (animal contaminated with feed) to the production or storage of the final product. Although the presence of mycotoxins in drycured meats has been reported in several regions of the world, the presence of these contaminants are not legislated in most countries. T…

Risk analysis of main mycotoxins occurring in food for children: An overview

Mycotoxins are secondary metabolites produced by fungi contaminating the food chain that are toxic to animals and humans. Children up to 12 years old are recognized as a potentially vulnerable subgroup with respect to consumption of these contaminants. Apart from having a higher exposure per kg body weight, they have a different physiology from that of adults. Therefore they may be more sensitive to neurotoxic, endocrine and immunological effects. For these reasons, a specific and up-to-date risk analysis for this category is of great interest. In this review, an accurate analysis of the main mycotoxins occurring in food intended for children (deoxynivalenol, aflatoxins, ochratoxins, patuli…

Bioactive compounds from mustard flours for the control of patulin production in wheat tortillas

Abstract Patulin (PAT) is a toxic fungal metabolite produced by Penicillium, Aspergillus and Byssochlamys growing especially in fruit and cereals. PAT exhibits a number of toxic effects in animals and its presence in food is undesirable. In this study the reduction of the mycotoxin PAT produced by a strain of Penicillium expansum, on wheat tortillas was studied using volatile bioactive compounds present in the oriental and yellow mustard flour and also using the standard solution of the antifungal compound allyl isothiocyanate (AIT), developing an active packaging with two different systems of release of those bioactive compounds. Also the kinetic of volatilization of the compounds used in …

Study of the chemical reduction of the fumonisins toxicity using allyl, benzyl and phenyl isothiocyanate in model solution and in food products

Abstract Fumonisins (FBs) are bioactive compounds produced by several strains of Fusarium spp. which contain a polyketide structure similar to sphinganine. These mycotoxins contain a free amino group that could work as an electron donor and react with the electrophile carbon present within the isothiocyanate (ITC) group. The objective of this study was to determine the effect of ITCs (allyl, benzyl and phenyl) on the stability of FB 1 , FB 2 and FB 3 . Firstly, PBS solutions at three pH levels (4, 7 and 9) were prepared and added with pairs of one FB (1 mg/L) plus one ITC (1 mg/L). Then, gaseous ITC was used to fumigate corn kernels and corn flour contaminated with FBs produced by Gibberell…

Immunotoxicological in vitro effects of BEA and ENN B on Jurkat cell line

Evaluation of immunologic effect of Enniatin A and quantitative determination in feces, urine and serum on treated Wistar rats.

Abstract Study of dietary supplementation with ENN A mycotoxin during 28 days of exposure time on Wistar rats to determinate its levels in serum, urine and feces and, to evaluate the immunologic effect in peripheral blood lymphocytes (PBL) is presented. The first method for ENN A extraction, determination and detection by LC–MS/MS in serum, urine and feces samples is reported. ENN A food dose administrated was detected in serum samples and influenced lymphocyte phenotyping. Levels in serum were founded from the second week of the experiment; reaching values of 4.76 μg/ml on the fourth week, which corresponds to 3.24 μg/ml in blood. PBL as T helper (CD4 + ) were presented in greater percenta…

Alternaria Mycotoxins in Food and Feed: An Overview

Alternariais one of the major mycotoxigenic fungal genera with more than 70 reported metabolites.Alternariamycotoxins showed notably toxicity, such as mutagenicity, carcinogenicity, induction of DNA strand break, sphingolipid metabolism disruption, or inhibition of enzymes activity and photophosphorylation. This review reports on the toxicity, stability, metabolism, current analytical methods, and prevalence ofAlternariamycotoxins in food and feed through the most recent published research. Half of the publications were focused on fruits, vegetables, and derived products—mainly tomato and apples—while cereals and cereal by-products represented 38%. The most studied compounds were alternario…

In vitro blood brain barrier exposure to mycotoxins and carotenoids pumpkin extract alters mitochondrial gene expression and oxidative stress.

Abstract Food and feed are daily exposed to mycotoxin contamination which effects may be counteracted by antioxidants like carotenoids. Some mycotoxins as well as carotenoids penetrate the blood brain barrier (BBB) inducing alterations related to redox balance in the mitochondria. Therefore, the in vitro BBB model ECV304 was subcultured for 7 days and exposed to beauvericine, enniatins, ochratoxin A, zearalenone (100 nM each), individually and combined, and pumpkin extract (500 nM). Reactive oxygen species were measured by fluorescence using the dichlorofluorescein diacetate probe at 0 h, 2 h and 4 h. Intracellular ROS generation reported was condition dependent. RNA extraction was performe…

Beauvericin and enniatin B effects on a human lymphoblastoid Jurkat T-cell model

Abstract Several mycotoxins exert their effect on the immunological system; some are classified as immunotoxic. Jurkat T-cells were used to study toxic effects of beauvericin (BEA) and enniatin B (ENN B). Both are not legislated mycotoxins with increasing presence in feed and food. Concentrations studied were from 1 to 15 μM at 24, 48 and 72 h. Cell death by increasing the percentage of apoptotic/necrotic cells was: BEA > ENN B. IC50 values ranged from 3 to 7.5 μM for BEA. ENN B 15 μM decreased viability (21-29%). The percentage of apoptotic/necrotic cells was BEA > ENN B at 24 h but not at 48 h. Caspase-3&7 activation profile varied, although both mycotoxins increased this activation. No d…

Enniatin A1, enniatin B1 and beauvericin on HepG2: Evaluation of toxic effects

Hepatotoxicity of three Fusarium mycotoxins, beauvericin (BEA) and two enniatins (ENNs) ENN A1 and ENN B1, in hepatocarcinoma cells (HepG2) were evaluated and compared. Concentrations used were 1.5 and 3 μM at 24, 48 and 72 h for each mycotoxin. Flow cytometry was used to examine enniatins effects on cell proliferation, to characterize the cell cycle phase where the cells blocked and to study the mitochondria role in ENNs-induced apoptosis. ENN B1 treated cells showed a time dependent G1 blockade at both concentrations used. ENN A1 and BEA decreased the apoptotic-necrotic percentage of cells comparing to control and disrupted the MMP as observed by TMRM and ToPro-3 fluorochromes signal. It …

In vivo toxicity studies of fusarium mycotoxins in the last decade: a review.

This review summarizes the information regarding the in vivo studies of Fusarium mycotoxins in the last decade. The most common studies are classified as subacute toxicity, subchronic toxicity, acute toxicity, toxicokinetic studies and teratogenicity in order of importance. The most used animals in in vivo studies are pigs, rats, chickens and mice. Fumonisin B1, deoxynivalenol, zearalenone, nivalenol and T-2 toxin are the most studied fusarotoxins. Studies with combinations of mycotoxins are also frequent, deoxynivalenol generally being one of them. The predominant route of administration is oral, administered mostly in the form of naturally contaminated feed. Other administration routes al…

Gaseous allyl isothiocyanate to inhibit the production of aflatoxins, beauvericin and enniatins by Aspergillus parasiticus and Fusarium poae in wheat flour

Abstract There is a growing concern about the presence of mycotoxins in foods, since up to 25% of cereals and cereal foods are contaminated with these compounds. Moreover, the general public is against the use of synthetic preservatives in foods and the use of natural antimicrobials in foods is a current trend. Allyl isothiocyanate (AITC) is a volatile antimicrobial derived from oriental and black mustard. The objective of this work was to evaluate the capacity of gaseous AITC in inhibiting the production of mycotoxins by Aspergillus parasiticus (aflatoxin producer) and Fusarium poae (beauvercin and enniatin producer) in wheat flour. Petri dish lids filled with 2 g of wheat flour were inocu…

Bioaccessibility Study of Aflatoxin B1 and Ochratoxin A in Bread Enriched with Fermented Milk Whey and/or Pumpkin

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 bio…

Multi-occurrence of twenty mycotoxinsin pasta and a risk assessment in the moroccan population

In the present study, the multi-occurrence of twenty (20) mycotoxins in pasta samples consumed in Morocco was assessed. For this, a modified Quick, Easy, Cheap Effective, Rugged, and Safe method was validated. The mycotoxins studied were identified and quantified by liquid chromatography&ndash

Nutrients associated with diseases related to aging: a new healthy aging diet index for elderly population

espanolIntroduccion: suelen usarse diversos indices dieteticos para medir la calidad de la nutricion en edades avanzadas. Sin embargo, ninguno de ellos fue disenado para evaluar la nutricion con el objetivo de evitar discapacidades en la poblacion anciana. Objetivos: identificar en la literatura cientifica los “nutrientes e ingestas” involucrados en el envejecimiento, asi como proponer un nuevo indice, considerando esta informacion, que sirva para evaluar la calidad de la nutricion con objeto de prevenir enfermedades relacionadas con el envejecimiento. Metodos: se realizo una revision bibliografica, obteniendo informacion sobre los nutrientes asociados con el envejecimiento. Todos estos nut…

Mitochondrial transcriptional study of the effect of aflatoxins, enniatins and carotenoids in vitro in a blood brain barrier model

C. maxima (var. Delica), a variety of pumpkin, is well known for its high concentration on carotenoids, possessing dietary benefits and antioxidant properties. Aflatoxins and enniatins are common mycotoxins present in food and feed with an extended toxicity profile in humans and animals. Both types of substances reach a wide range of tissues and organs and have the capability to penetrate the blood brain barrier. Since carotenoids and mycotoxins have been reported to modify diverse mitochondrial processes individually, transcriptional in vitro studies on human epithelial cells ECV 304 were conducted to analyze the relative expression of 13 mitochondria related genes. ECV 304 cells were diff…

Proteomics evaluation of enniatins acute toxicity in rat liver

Abstract Enniatins (ENs) are emerging mycotoxins produced by Fusarium fungi which are cytotoxic also at low concentrations due to its ionophoric properties. The aim of this study was to evaluate the hepatic toxicity of ENs exposure at different concentrations in Wistar rats through a proteomic approach. Animals were intoxicated by oral gavage with medium (EN A 256, ENA1 353, ENB 540, ENB1 296 μg/mL) and high concentrations (ENA 513, ENA1 706, ENB 1021, ENB1 593 μg/mL) of an ENs mixture and sacrificed after 8 h. Protein extraction was performed using powdered liver. Peptides were analyzed using a liquid chromatography coupled with a quadrupole time-of-flight mass spectrometer. Proteins were …

Adherence to dietary treatment and clinical factors associated with anti-transglutaminase antibodies in celiac disease during the follow-up

Introduction In clinical practice, celiac disease (CD) is monitored through anti-transglutaminase (TGA-IgA) antibody levels. The normalization of serum levels in successive periodic measurements indicates good response and adherence to dietary treatment. Objectives To evaluate the factors associated with the evolution of TGA-IgA antibodies and their association with dietary non-compliance and diseases related to CD. Methods This prospective observational study was carried out in 254 participants, who were recruited from patients from a hospital in southern Spain. Information about sex, age, serological test results, HLA DQ2/DQ8 haplotypes, mucosal atrophy, gastrointestinal and extra-intesti…

Geographic conditioning in dietary, social, and health patterns in elderly population with disabilities

Abstract Objective The aim of this study was to identify the most relevant variables defining the dietary, social, and health patterns of elderly populations with disabilities, considering their geographic profile. Methods A cross-sectional study was carried out in a sample of 354 disabled, free-living elderly adults from three different geographic profiles (metropolitan, rural, and mixed profile). The dietary data were obtained through a validated food habit questionnaire. The data regarding health status, cohabitation unit, and social benefits were obtained through the public social services. A standardized principal component analysis was used to select the most relevant variables, by co…

Mycotoxin contamination in laboratory rat feeds and their implications in animal research

Compound feed is particularly vulnerable to multi-mycotoxin contamination. A method for the determination of 12 mycotoxins; enniatins A, A1, B, B1; aflatoxins B1, B2, G1, G2; OTA; ZEA; T-2 and HT-2 by liquid chromatography-tandem mass spectrometry has been developed and applied for the analysis of laboratory rat commercial feeds. The method trueness was checked by recovery assays at three different spiked levels (n = 9). Recoveries ranged from 73% to 112%, and the intra-day and inter-day precision were lower than 9% and 13%, respectively. Limits of quantitation were lower than 15 μg/kg. Twenty-seven laboratory rats feed samples showed multi-contamination by at least three up to six differen…

Involvement of enniatins-induced cytotoxicity in human HepG2 cells.

Enniatins (ENNs) are mycotoxins found in Fusarium fungi and they appear in nature as mixtures of cyclic depsipeptides. The ability to form ionophores in the cell membrane is related to their cytotoxicity. Changes in ion distribution between inner and outer phases of the mitochondria affect to their metabolism, proton gradient, and chemiosmotic coupling, so a mitochondrial toxicity analysis of enniatins is highly recommended because they host the homeostasis required for cellular survival. Two ENNs, ENN A and ENN B on hepatocarcinoma cells (HepG2) at 1.5 and 3 μM and three exposure times (24, 48 and 72 h) were studied. Flow cytometry was used to examine their effects on cell proliferation, t…

Binary and tertiary combination of alternariol, 3-acetyl-deoxynivalenol and 15-acetyl-deoxynivalenol on HepG2 cells: Toxic effects and evaluation of degradation products.

Fungi producers of mycotoxins are able to synthesize more than one toxin. Alternariol (AOH) is one of the mycotoxins produced by several Alternaria species, the most common one being Alternaria alternata. The toxins 3-Acetyl-deoxynivalenol (3-ADON) and 15-Acetyl-deoxynivalenol (15-ADON) are acetylated forms of deoxynivalenol (DON) produced by Fusarium graminearum. In the present work it is determined and evaluated the toxic effects of binary and tertiary combination treatment of HepG2 cells with AOH, 3-ADON and 15-ADON, by using the MTT assay (3-[4,5-dimethylthiazol-2-yl]-2,5 diphenyl tetrazolium bromide), to subsequently apply the isobologram method and elucidate if the mixtures of these m…

Risk assessment of mycotoxins in coffee beverages

Effect of different thermal processes in the reduction of enniatins in fish tissues

Influence of the antimicrobial compound allyl isothiocyanate against the Aspergillus parasiticus growth and its aflatoxins production in pizza crust.

Abstract Aflatoxins (AFs) are secondary metabolites produced by different species of Aspergillus, such as Aspergillus flavus and Aspergillus parasiticus, which possess mutagenic, teratogenic and carcinogenic activities in humans. In this study, active packaging devices containing allyl isothiocyanate (AITC) or oriental mustard flour (OMF) + water were tested to inhibit the growth of A. parasiticus and AFs production in fresh pizza crust after 30 d. The antimicrobial and anti-aflatoxin activities were compared to a control group (no antimicrobial treatment) and to a group added with commercial preservatives (sorbic acid + sodium propionate). A. parasiticus growth was only inhibited after 30 d…

Effect of allyl isothiocyanate on transcriptional profile, aflatoxin synthesis, and Aspergillus flavus growth.

Abstract The goals of this study were to determine the efficacy of allyl isothiocyanate (AITC) against the growth of A. flavus and Aflatoxin B1 (AFB1) production as well as to evaluate changes in the transcriptome profile when colonizing maize. A. flavus was inoculated in potato dextrose agar (PDA), the plates were placed inside glass jars and the mycelial growth (MG) was monitored for 7 d. Likewise, maize grains were contaminated with A. flavus in glass jars of 1 L and treated with 0.125, 0.25, 0.5, 1 and 5 µL of AITC. The moisture content (MC) of grains was 15 and 21%. After 7 days of storage, the MG was significantly reduced in doses higher than 0.125 µL/L of AITC. All doses of AITC redu…

The role of pumpkin pulp extract carotenoids against mycotoxin damage in the blood brain barrier

Some mycotoxins such as beauvericin (BEA), ochratoxin A (OTA), and zearalenone (ZEA) can cross the blood brain barrier, which is why we tested the anti-inflammatory action of a pumpkin carotenoid extract (from the pulp) against these mycotoxins and their combinations (OTA+ZEA and OTA+ZEA+BEA) on a blood brain barrier model with co-cultured ECV304 and C6 cells using an untargeted metabolomic approach. The cells were added with mycotoxins at a concentration of 100 nmol/L per mycotoxin and pumpkin carotenoid extract at 500 nmol/L. For control we used only vehicle solvent (cell control) or vehicle solvent with pumpkin extract (extract control). After two hours of exposure, samples were analysed…

Applications of flow cytometry to toxicological mycotoxin effects in cultured mammalian cells: a review.

This review gives an overview of flow cytometry applications to toxicological studies of several physiological target sites of mycotoxins on different mammalian cell lines. Mycotoxins are secondary metabolites of fungi that may be present in food, feed, air and water. The increasing presence of mycotoxins in crops, their wide distribution in the food chain, and their potential for toxicity demonstrate the need for further knowledge. Flow cytometry has become a valuable tool in mycotoxin studies in recent years for the rapid analysis of single cells in a mixture. In toxicology, the power of these methods lies in the possibility of determining a wide range of cell parameters, providing valuab…