Co-occurrence Of Mycotoxins Research Articles

Risk of dietary and breastmilk exposure to mycotoxins among lactating women and infants 2-4 months in northern India.

Mycotoxins are carcinogenic secondary metabolites of fungi that have been linked to infant growth faltering. In this study, we quantified co‐occurring mycotoxins in breast milk and food samples from Haryana, India, and characterized determinants of exposure. Deterministic risk assessment was conducted for mothers and infants. We examined levels of eight mycotoxins (Aflatoxin B1, B2, G1, G2, M1, M2; Ochratoxin A, B) in 100 breast milk samples (infants 2–4 months) using ultra‐high‐performance liquid chromatography tandem mass spectrometry. Aflatoxin B1 (AFB1), fumonisin B1 (FB1) and deoxynivalenol (DON) were detected in several food items (n = 298) using enzyme‐linked immunosorbent assays. We report novel data on the presence of mycotoxins in breast milk samples from India. Whereas breast milk concentrations (AFM1 median: 13.7; range: 3.9–1200 ng/L) remain low, AFM1 was detected above regulatory limits in 27% of animal milk samples. Additionally, 41% of infants were above provisional maximum tolerable daily intake (PMTDI) limits for AFM1 due to consumption of breast milk (mean: 3.04, range: 0.26–80.7 ng kg−1 bw day−1). Maternal consumption of breads (p < 0.05) was associated with breast milk AFM1 exposure. AFB1 (μg/kg) was detected in dried red chilies (15.7; 0–302.3), flour (3.13; 0–214.9), groundnuts (0; 0–249.1), maize (56.0; 0–836.7), pearl millet (1.85; 0–160.2), rice (0; 0–195.6), wheat (1.9; 0–196.0) and sorghum (0; 0–63.5). FB1 (mg/kg) was detected in maize (0; 0–61.4), pearl millet (0; 0–35.4) and sorghum (0.95; 0–33.2). DON was not detected in food samples. Mothers in our study exceeded PMTDI recommendations for AFB1 due to consumption of rice and flour (mean: 75.81; range: 35.2–318.2 ng kg−1 bw day−1). Our findings show the presence of Aflatoxin B1 and M1 at various levels of the food chain and in breast milk, with estimated intakes exceeding PMTDI recommendations. Aflatoxins are known carcinogens and have also been linked to stunting in children. Their presence across the food system and in breast milk is concerning, thus warranting further research to replicate and expand on our findings and to understand implications for maternal and child health.

Co-Occurrence and Levels of Mycotoxins in Fish Feeds in Kenya.

This study determined the presence, levels and co-occurrence of mycotoxins in fish feeds in Kenya. Seventy-eight fish feeds and ingredients were sampled from fish farms and fish feed manufacturing plants and analysed for 40 mycotoxins using high-performance liquid chromatography-high resolution mass spectrometry. Twenty-nine (73%) mycotoxins were identified with 76 (97%) samples testing positive for mycotoxins presence. Mycotoxins with the highest prevalences were enniatin B (91%), deoxynivalenol (76%) and fumonisin B1 (54%) while those with the highest maximum levels were sterigmatocystin (<30.5–3517.1 µg/kg); moniliformin (<218.9–2583.4 µg/kg) and ergotamine (<29.3–1895.6 µg/kg). Mycotoxin co-occurrence was observed in 68 (87%) samples. Correlations were observed between the fumonisins; enniatins B and zearalenone and its metabolites. Fish dietary exposure estimates ranged between <0.16 and 43.38 µg/kg body weight per day. This study shows evidence of mycotoxin presence and co-occurrence in fish feeds and feed ingredients in Kenya. Fish exposure to these levels of mycotoxins over a long period of time may lead to adverse health effects due to their possible additive, synergistic or antagonist toxic effects. Measures to reduce fish feed mycotoxin contamination should be taken to avoid mycotoxicosis in fish and subsequently in humans and animals through residues.

Quantification of multi-mycotoxin in cereals (maize, maize porridge, sorghum and wheat) from Limpopo province of South Africa

ABSTRACT Mycotoxins are secondary metabolites that are produced by filamentous mycotoxigenic fungi belonging to the Alternaria, Aspergillus, Fusarium and Penicillium genera amongst others. Multi-class mycotoxins were extracted from 55 cereal samples and analysed using liquid chromatography-tandem mass spectrometry. The adopted extraction method for maize, maize porridge, sorghum and wheat was comprehensively validated. This method was further tested to determine the natural occurrence of mycotoxins in foodstuffs. Twelve (12) out of 22 mycotoxins were detected in maize, maize porridge, sorghum and wheat, including α-zearalenol (α-ZEL) (89%), fumonisin B3 (FB3) (84%), fumonisin B1 (FB1) (80%), tenuazonic acid (TeA) (78%), ochratoxin B (42%), deoxynivalenol (DON) (12%), ochratoxin A (11%), 3-acetyldeoxynivalenol (7%), sterigmatocystin (STG) (6%), 15-acetyldeoxynivalenol (2%), cyclopiazonic acid (CPA) (2%) and aflatoxin B2 (2%). The data revealed high incidence rate of α-ZEL (range: 6.5–70.5 µg kg−1) in all matrices. Maize samples had high mycotoxin co-occurrence compared to other matrices. All recovered mycotoxins in food commodities were within the maximum regulatory limits, with the exception of fumonisins (FB1 and FB3) exceeded the South African and European Commission regulation, and the highest concentration was 2153 µg kg−1 in maize. It is essential to monitor the level of emerging mycotoxins in food commodities from rural areas as trace amount of CPA (< limit of quantification), STG (range: 0.30–0.74 µg kg−1) were detected and high concentration of TeA (292.7 µg kg−1) was detected in sorghum. The occurrence of these mycotoxins further encourages frequent analyses, their co-occurrence in the samples poses a significant threat to public health and more emphasis should thus be placed on reducing the contamination levels of these toxins in staples.

Mycotoxins and flours: Effect of type of crop, organic production, packaging type on the recovery of fungal genus and mycotoxins

Heat-stable mycotoxins are widely detected in flour and produced by Aspergillus spp., Fusarium spp. and Penicillium spp. Forty different flours purchased in Italy are used to assess potential risk factors via a systematically screening of a number of variables: the type of flour, organic, whole and white wheat, types of packaging (paper, plastic and weight). Fungal recovery and co-occurrence of specific mycotoxins was also assessed. The results showed that flour originated from fruits had a significant higher recovery of fungi, while seed/pseudocereals had the highest mycotoxins detection. Flours originating from organic agriculture are more prone to higher fungal recovery and mycotoxins detection when compared with not-organic flours. Packaging is also important: packaging weighting less than 376 g supports significantly more fungal recovery and the plastic packages was observed to retain more fungi and mycotoxins detection when compared with paper. Recovery measured as Log (CFU/g) of fungal genera is not directly proportional to the amount of mycotoxins. Finally, linear regression and mixed logit regression models show that the mean level of aflatoxins B1 (ng/g on the logarithmic scale) reduces by 0.485 when moving from an organic to a non-organic flour, while a significant increase of 0.369 when moving from paper to a plastic packaging.

Mass spectrometry-based detection and risk assessment of mycotoxin contamination of ‘kankankan’ used for roasted meat consumption in Abidjan, Côte d’Ivoire

ABSTRACT ‘Kankankan’ is a popular spice powder used to season roasted meat in Côte d’Ivoire. However, produced in a traditional way, the conditions of production and storage of kankankan favour the proliferation of mycotoxin-producing fungal strains. The aim of this study was to carry out an inventory of mycotoxin contamination of this spice powder and to assess risk exposure to consumers. In total, 75 samples of kankankan were collected from wholesalers (6), sellers of kankankan in the markets (35) and sellers of roasted meat (34) across three municipalities of Abidjan, the economic capital of Côte d’Ivoire. Ultra-high-performance liquid chromatography-tandem mass spectrometry (UHPLC-MS/MS) was used to detect and quantify nine different mycotoxins. Dietary exposure was calculated by using estimated daily intake (EDI), whereas risk characterisation was assessed using the margin of exposure (MOE) approach. Aflatoxins and fumonisins were found in 99% of samples assessed, while contamination with beauvericin was proportionally lowest (28%). At all the three types of actors within the food production chain (wholesalers, kankankan sellers and roasted meat sellers) the mean concentrations of aflatoxin B1 (AFB1) in samples exceeded the European standard for spice mixtures, with concentrations reaching up to 502 µg/kg. The estimated daily intakes of aflatoxins observed in the different populations were above the recommended level of 0.017 ng kg−1 b.w. day−1. The MOES values for adolescents and adults were 8.10 and 12.78, respectively, well below the safe margin of 10,000. The co-occurrence of mycotoxins in kankankan samples together with high aflatoxin exposure to consumers represent a potential risk to public health, calling for immediate risk management and education of kankankan producers and consumers.

Las micotoxinas: el enemigo silencioso

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.

Study on mycotoxin contamination of maize kernels in Spain

Mycotoxins are secondary metabolites produced mainly by fungal species belonging to the genera Fusarium, Aspergillus and Penicillium and belong to the most relevant contaminants of food and feed. Cereals are the main source of mycotoxins in the diet. The most prominent mycotoxins are aflatoxins B1, B2, G1 and G2 (AFB1, AFB2, AFG1 and AFG2), fumonisins B1 and B2 (FB1 and FB2), ochratoxin A (OTA), zearalenone (ZEA), deoxynivalenol (DON), 3- and 15-acetyl-deoxynivalenol (3- and 15-ADON), and T-2 and HT-2 toxins. Maximum levels allowed in food are very different depending on mycotoxin and food type, consumer susceptibility and current legislation in each country. Among cereals, maize and its derivatives deserve special attention. The present study was designed to investigate the occurrence of these mycotoxins in Spanish maize kernels collected in different regions for a five-year period (2015–2019) using a multi-mycotoxin UPLC-MS/MS method. Ninety-eight samples of maize kernels collected in 26 Spanish grain stores were ground to fine powder, extracted with acetonitrile-water-formic acid (80:19:1, v/v/v), centrifuged and analyzed by UPLC-ESI-MS/MS using matrix-matched calibration. The method was validated and mean recovery rates were 83–103.4% and mean relative standard deviations ranged from 4.1 to 8.5%. FB1, FB2, DON, 3-ADON, ZEA, AFB1, AFB2, AFG2, T-2 and HT-2 toxins were detected at levels higher than their respective limits of quantification in 71.4%, 56.2%, 31.6%, 5.02%, 24.5%, 9.2%, 9.2%, 2%, 5.1% and 5.1% of maize kernel samples, respectively. OTA was not detected. Among the positive samples, 20.4%, 5.1%, 3%, and 3% had levels exceeding the European Union permissible limits for FB1+FB2, ZEA, AFB1, and total aflatoxins, respectively. Co-occurrence of various mycotoxins (two to seven) in the same sample at quantifiable levels was observed in 33.5% of samples being the association of FB1, FB2 and DON the most frequent, followed by the presence of FB1, FB2, ZEA and DON.

Occurrence and co-occurrence of mycotoxins in apple and apple products from China

Mycotoxins are abiotic hazards produced by certain fungi that can grow on apples. Consequently, their prevalence in apple and appple products may be relatively high. In this study, 205 healthy apples and 104 rotten apples from three provinces of China (Liaoning, Hebei and Shandong), as well as 107 apple products purchased from Chinese local and online markets, were used to analyze 7 mycotoxins. The results showed that none of the mycotoxins were found in healthy apples. Six mycotoxins (patulin, alternariol, alternariol monomethyl ether, altenuene, tentoxin and tenuazonic acid) were detected in rotten apples and apple products, and patulin was the most common and frequent one. Contamination levels of patulin, altenuene, tentoxin and tenuazonic acid were significant difference in different parts of rotten apples among three provinces. Mycotoxin co-occurrence was common in rotten apples and apple products, and the rates of rotten apples and apple products contaminated by two or more mycotoxins were 73.08% and 4.67%, respectively.

Mycotoxins in maize: mitigation actions, with a chain management approach

Maize is the principal staple food/feed crop exposed to mycotoxins, and the co-occurrence of multiple mycotoxins and their metabolites has been well documented. This review presents the infection cycle, ecology, and plant-pathogen interactions of Aspergillus and Fusarium species in maize, and current knowledge on maize chain management to mitigate the occurrence of aflatoxins and fumonisins. Preventive actions include at pre-harvest, as part of cropping systems, at harvest, and at post-harvest, through storage, processing, and detoxification to minimize consumer exposure. Preventive actions in the field have been recognized as efficient for reducing the entrance of mycotoxins into production chains. Biological control of Aspergillus flavus has been recognized to minimize contamination with aflatoxins. Post-harvest maize grain management is also crucial to complete preventive actions, and has been made mandatory in government food and feed legislation.

Occurrence and Co-Occurrence of Mycotoxins in Cereal-Based Feed and Food.

Dietary (co)-exposure to mycotoxins is associated with human and animal health concerns as well as economic losses. This study aims to give a data-based insight from the scientific literature on the (co-)occurrence of mycotoxins (i.e., parent and modified forms) in European core cereals, and to estimate potential patterns of co-exposure in humans and animals. Mycotoxins were mainly reported in wheat and maize showing the highest concentrations of fumonisins (FBs), deoxynivalenol (DON), aflatoxins (AFs), and zearalenone (ZEN). The maximum concentrations of FB1+FB2 were reported in maize both in feed and food and were above legal maximum levels (MLs). Similar results were observed in DON-food, whose max concentrations in wheat, barley, maize, and oat exceeded the MLs. Co-occurrence was reported in 54.9% of total records, meaning that they were co-contaminated with at least two mycotoxins. In the context of parental mycotoxins, co-occurrence of DON was frequently observed with FBs in maize and ZEN in wheat; DON + NIV and DON + T2/HT2 were frequently reported in barley and oat, respectively. Apart from the occurrence of ZEN and its phase I and phase II modified forms, only a limited number of quantified data were available for other modified forms; i.e., mainly the acetyl derivatives of DON. Data gaps are highlighted together with the need for monitoring studies on multiple mycotoxins to identify co-occurrence patterns for parent mycotoxins, metabolites, and their modified forms.

Simultaneous Direct Analysis of Aflatoxins and Ochratoxin A in Cereals and Their Processed Products by Ultra-High Performance Liquid Chromatography with Fluorescence Detection

Abstract Background: Mycotoxins such as aflatoxins (AFs) and ochratoxin A (OTA) can pose severe health hazards because of their toxicity. Given a wide range of food matrices susceptible to fungal infections and possible cooccurrence of mycotoxins at different concentrations, validated multimycotoxin and multimatrix methods are strongly warranted. Objective: The aim of this research was to develop a simple and fast ultra-high performance LC (UHPLC) fluorescence detection (FLD)–based method to simultaneously determine AFs (B1, G1, B2, and G2) and OTA and, furthermore, to carry out single-laboratory validation in a range of cereals and processed product matrices. Methods: The sample preparation involved homogenization and extraction with methanol–water (80 + 20). For cleanup, an aliquot (3 mL) was diluted with phosphate-buffered saline, loaded on an immunoaffinity column (AFLAOCHRA PREP®), and eluted with methanol (1 mL). The cleaned extract was diluted with 0.2% acetic acid (at a 1:1 ratio) before injection into an ultra-high performance liquid chromatograph. To perform simultaneous analysis of AFs and OTA, the FLD program was developed by switching the excitation wavelength in a single chromatographic run. Results: The method provided LOQs of 0.25 and 1 ng/g for AFs and OTA, respectively, without involving any derivatization. In rice, the recoveries of AFs ranged from 84 to 106%, whereas OTA had a recovery above 72%, with the repeatability relative SDs &amp;lt;12% for both analytes. The method was successfully applied to a range of naturally contaminated market samples. Conclusions: The method is suitable for regulatory testing because of its significant time and cost effectiveness and sensitivity in compliance with the regulatory maximum levels. Highlights: The study achieves high-throughput analysis of AFs and OTA in raw and processed cereals using simultaneous extraction, cleanup, and UHPLC-FLD. Method sensitivity complies with the regulatory maximum levels. Single-laboratory validation results meet analytical QC requirements.

Efficacy of a Yeast Cell Wall Extract to Mitigate the Effect of Naturally Co-Occurring Mycotoxins Contaminating Feed Ingredients Fed to Young Pigs: Impact on Gut Health, Microbiome, and Growth.

Mycotoxins are produced by fungi and are potentially toxic to pigs. Yeast cell wall extract (YCWE) is known to adsorb mycotoxins and improve gut health in pigs. One hundred and twenty growing (56 kg; experiment 1) and 48 nursery piglets (6 kg; experiment 2) were assigned to four dietary treatments in a 2 × 2 factorial design for 35 and 48 days, respectively. Factors were mycotoxins (no addition versus experiment 1: 180 μg/kg aflatoxins and 14 mg/kg fumonisins; or experiment 2: 180 μg/kg aflatoxins and 9 mg/kg fumonisins, and 1 mg/kg deoxynivalenol) and YCWE (0% versus 0.2%). Growth performance, blood, gut health and microbiome, and apparent ileal digestibility (AID) data were evaluated. In experiment 1, mycotoxins reduced ADG and G:F, and duodenal IgG, whereas in jejunum, YCWE increased IgG and reduced villus width. In experiment 2, mycotoxins reduced BW, ADG, and ADFI. Mycotoxins reduced ADG, which was recovered by YCWE. Mycotoxins reduced the AID of nutrients evaluated and increased protein carbonyl, whereas mycotoxins and YCWE increased the AID of the nutrients and reduced protein carbonyl. Mycotoxins reduced villus height, proportion of Ki-67-positive cells, and increased IgA and the proportion of bacteria with mycotoxin-degrading ability, whereas YCWE tended to increase villus height and reduced IgA and the proportion of pathogenic bacteria in jejunum. The YCWE effects were more evident in promoting gut health and growth in nursery pigs, which showed higher susceptibility to mycotoxin effects.

Evaluation of High-Resolution Mass Spectrometry for the Quantitative Analysis of Mycotoxins in Complex Feed Matrices

The selective and sensitive analysis of mycotoxins in highly complex feed matrices is a great challenge. In this study, the suitability of OrbitrapTM-based high-resolution mass spectrometry (HRMS) for routine mycotoxin analysis in complex feeds was demonstrated by the successful validation of a full MS/data-dependent MS/MS acquisition method for the quantitative determination of eight Fusarium mycotoxins in forage maize and maize silage according to the Commission Decision 2002/657/EC. The required resolving power for accurate mass assignments (<5 ppm) was determined as 35,000 full width at half maximum (FWHM) and 70,000 FWHM for forage maize and maize silage, respectively. The recovery (RA), intra-day precision (RSDr), and inter-day precision (RSDR) of measurements were in the range of 94 to 108%, 2 to 16%, and 2 to 12%, whereas the decision limit (CCα) and the detection capability (CCβ) varied from 11 to 88 µg/kg and 20 to 141 µg/kg, respectively. A set of naturally contaminated forage maize and maize silage samples collected in northern Germany in 2017 was analyzed to confirm the applicability of the HRMS method to real samples. At least four Fusarium mycotoxins were quantified in each sample, highlighting the frequent co-occurrence of mycotoxins in feed.

Occurrence of Mycotoxins in Extruded Commercial Cat Food.

The occurrence of the most important mycotoxins (deoxynivalenol, fumonisin B1 and B2, aflatoxins B1, B2, G1, and G2, ochratoxin A, zearalenone, T-2, and HT-2 toxins) was determined in 64 extruded cat foods purchased in Italy through ultra-performance liquid chromatography coupled with tandem mass spectrometry. Deoxynivalenol and fumonisins were the most common contaminants (quantified in 80 and 95% of the samples, respectively). Conversely, aflatoxins B2, G1, and G2 were not identified in any sample. Some cat foods exceeded the regulatory limit for aflatoxin B1 (n = 3) or the guidance values for zearalenone (n = 3), fumonisins (n = 2), ochratoxin A (n = 1), and T-2 (n = 1) recently established for pets in the European Union. A widespread co-occurrence of mycotoxins was observed (28, 42, and 8% of the samples contained quantifiable amounts of two, three, and four mycotoxins, respectively). This study describes criticisms regarding the mycotoxin issue in pet food and suggests an improvement of the monitoring of the pet food chain.

Target Analysis and Retrospective Screening of Multiple Mycotoxins in Pet Food Using UHPLC-Q-Orbitrap HRMS.

A comprehensive strategy combining a quantitative method for 28 mycotoxins and a post-target screening for other 245 fungal and bacterial metabolites in dry pet food samples were developed using an acetonitrile-based extraction and an ultrahigh-performance liquid chromatography coupled to high-resolution mass spectrometry (UHPLC-Q-Orbitrap HRMS) method. The proposed method showed satisfactory validation results according to Commission Decision 2002/657/EC. Average recoveries from 72 to 108% were obtained for all studied mycotoxins, and the intra-/inter-day precision were below 9 and 14%, respectively. Results showed mycotoxin contamination in 99% of pet food samples (n = 89) at concentrations of up to hundreds µg/kg, with emerging Fusarium mycotoxins being the most commonly detected mycotoxins. All positive samples showed co-occurrence of mycotoxins with the simultaneous presence of up to 16 analytes per sample. In the retrospective screening, up to 54 fungal metabolites were tentatively identified being cyclopiazonic acid, paspalitrem A, fusaric acid, and macrosporin, the most commonly detected analytes.

A review on combined effects of moniliformin and co-occurringFusarium toxins in farm animals

Co-occurrence of mycotoxins in food and feed represents the rule rather than the exception. Information about combinatory toxic effects of co-occurring mycotoxins is scarce, in particular the effects that mixtures of mycotoxins in feed may have on farm animals. This review focusses on studies on the combined effects of moniliformin and co-occurring mycotoxins in feed on farm animals. Moniliformin is a mycotoxin of emerging scientific interest, which may co-occur with many other mycotoxins, especiallyFusarium mycotoxins. Oral exposure to moniliformin reduces feed consumption and body weight gain in poultry, in pigs and catfish, and induces cardiotoxic effects and/or alterations in serum biochemical and haematological parameters. In this review only experiments comparing effects as a result of the exposure to a combination of mycotoxins with effects due to the exposure to single mycotoxins were considered. Identified published studies on combined toxicity have been limited to combinations of moniliformin with either fumonisin B1 or deoxynivalenol, and were performed with poultry, pigs, and catfish. Most of the moniliformin/fumonisin B1 investigations involved poultry and focussed on adverse effects on feed intake, weight gain and immune response, as well as organ lesions. These studies mainly reported an interactive toxicity of moniliformin and fumonisin B1 but did not allow identification of the type of interaction. Likewise, no indication could be given for the interaction detected for both mycotoxins on weight gains of catfish. For the moniliformin/deoxynivalenol combination, only one study with broiler chickens was found relevant. This study concluded additive or less than additive toxicity, using kidney lesions and renal tubular epithelial degeneration as endpoints. While possible interactions between moniliformin and fumonisin B1 or deoxynivalenol were identified, the conclusions are based on limited studies and experimental designs. Further studies on the combined toxicity of moniliformin with other mycotoxins and other animal species would be needed.

Fungal and mycotoxin problems in grape juice and wine industries

The fungal diseases in the vineyard reduce grape quality influencing the volatile profile and consequently impair the aroma, taste, and color of juice and wine. In addition to this sensory effect, the occurrence of filamentous fungi in grapes may result in mycotoxins, including alternariol, aflatoxins, tenuazonic acid, fumonisin B2, ochratoxin A (OTA), patulin citrinin, among others. OTA is the most studied mycotoxin in grape products and can effectively represent a concern for industries. This short review discloses that promising results have been obtained with the use of antagonistic microorganisms or natural products for the toxigenic fungi control in the vineyard. Reduction of OTA levels throughout the production stages of juice and wine is well documented in the literature. However, studies taking into account the impacts of climate change on vine and the resilience of toxigenic fungi are still a challenge. In addition, the co-occurrence of OTA (including modified forms) and other mycotoxins in grapes products of different cultivars and regions needs to be evaluated to verify the risk of human exposure to these compounds.

Simultaneous Direct Analysis of Aflatoxins and Ochratoxin A in Cereals and Their Processed Products by Ultra-High Performance Liquid Chromatography with Fluorescence Detection.

Background: Mycotoxins such as aflatoxins (AFs) and ochratoxin A (OTA) can pose severe health hazards because of their toxicity. Given a wide range of food matrices susceptible to fungal infections and possible cooccurrence of mycotoxins at different concentrations, validated multimycotoxin and multimatrix methods are strongly warranted. Objective: The aim of this research was to develop a simple and fast ultra-high performance LC (UHPLC) fluorescence detection (FLD)-based method to simultaneously determine AFs (B1, G1, B2, and G2) and OTA and, furthermore, to carry out single-laboratory validation in a range of cereals and processed product matrices. Methods: The sample preparation involved homogenization and extraction with methanol-water (80 + 20). For cleanup, an aliquot (3 mL) was diluted with phosphate-buffered saline, loaded on an immunoaffinity column (AFLAOCHRA PREP®), and eluted with methanol (1 mL). The cleaned extract was diluted with 0.2% acetic acid (at a 1:1 ratio) before injection into an ultra-high performance liquid chromatograph. To perform simultaneous analysis of AFs and OTA, the FLD program was developed by switching the excitation wavelength in a single chromatographic run. Results: The method provided LOQs of 0.25 and 1 ng/g for AFs and OTA, respectively, without involving any derivatization. In rice, the recoveries of AFs ranged from 84 to 106%, whereas OTA had a recovery above 72%, with the repeatability relative SDs <12% for both analytes. The method was successfully applied to a range of naturally contaminated market samples. Conclusions: The method is suitable for regulatory testing because of its significant time and cost effectiveness and sensitivity in compliance with the regulatory maximum levels. Highlights: The study achieves high-throughput analysis of AFs and OTA in raw and processed cereals using simultaneous extraction, cleanup, and UHPLC-FLD. Method sensitivity complies with the regulatory maximum levels. Single-laboratory validation results meet analytical QC requirements.

Natural Occurrence and Co-Contamination of Twelve Mycotoxins in Industry-Submitted Cool-Season Cereal Grains Grown under a Low Heat Unit Climate Condition.

This study aims to evaluate the prevalence of mycotoxins in industry-submitted cool-season barley and wheat grown under low heat unit climate conditions. Seventy-two barley samples and 83 wheat samples were submitted by producers and industry from May 2016 to May 2017. The concentrations of twelve common mycotoxins, including nivalenol (NIV), deoxynivalenol (DON), 3-acetyldeoxynivalenol (3-ADON), 15-acetyldeoxynivalenol (15-ADON), ochratoxin A (OTA), zearalenone (ZEN), α-zearalenol (α-ZAL), β-zearalenol (β-ZAL), diacetoxyscirpenol (DAS), T-2 toxin (T-2), HT-2 toxin (HT-2), and aflatoxin B1 (AFB1), were determined using the liquid chromatography/tandem mass spectrometry method. Mycotoxins were detected in 40 barley (56%) and 35 wheat (42%) samples submitted by producers and industry. DON showed the highest incidence in barley (44%) and wheat (33%). None of the barley samples contained detectable DAS and no wheat samples tested positive for α-ZAL, DAS, T-2, or AFB1. Co-occurrence of DON and other mycotoxins was frequently observed. Among the mycotoxin-positive samples, 70% of barley samples and 54% of wheat samples were co-contaminated with at least two mycotoxins. Four barley (6%) and five wheat (6%) samples contained levels of DON above 1000 μg/kg (regulatory level in diets for lactating dairy animals) and HT-2 content in five barley (7%) and four wheat (5%) samples exceeded 100 μg/kg (regulatory level in diets for cattle and poultry). Overall, contamination of these mycotoxins was more frequent and more severe in barley in comparison with wheat that was submitted by producers and industry. Comprehensive strategies, including the prevention of Fusarium toxins contamination, the routine monitoring of their prevalence, the detoxification of them in feed and food, as well as the inhibition of their absorption in the gastrointestinal tract, are highly required. A rapid detection method needs to be developed to screen mycotoxins in industry-submitted cool-season cereal grains.

Co-occurrence of mycotoxins, including aflatoxins, trichothecenes, and zearalenone, in Chinese feedstuffs collected in 2013 and 2014

A total of 357 feedstuff samples, including cottonseed meal plus soybean meal (CSM), wheat bran (WB) and dried distillers grain with solubles (DDGS) collected from 14 provinces in China in 2013 and 2014 were analysed for the co-occurrence of mycotoxins, including 4 aflatoxins, 7 trichothecenes, and zearalenone (ZEA), by high-performance liquid chromatography/electrospray ionization tandem mass spectrometry (HPLC/ESI-MS/MS). Deoxynivalenol (DON) was the most prevalent mycotoxin found in Chinese feedstuffs, followed by ZEA and aflatoxin B1 (AFB1). It was found that out of 357 samples, 27 (7.6%), 11 (3.1%) and 5 (1.4%) samples were positive for DON, ZEA and AFB1 at levels exceeding the Chinese regulatory limit of 1000 μg/kg, 500 μg/kg, 50 μg/kg for DON, ZEA, and AFB1, respectively. DON, ZEA, and AFB1 levels ranged from 1,005.5 to 2,893.1 μg/kg, 510.8 to 4,845.9 μg/kg, and 53.9 to 72.9 μg/kg, respectively. These samples were more easily co-contaminated by other mycotoxins, such as DON, ZEA, AFB1, AFB2, 3-acetyldeoxynivalenol (3-ADON), and 15-ADON. CSM was more easily contaminated by aflatoxins than WB and DDGS, and WB was more easily contaminated by DON and ZEA. Besides, 339 samples (95.0%, 339/357) were contaminated by at least two mycotoxins, and 16 samples (4.5%, 16/357) were only contaminated by one mycotoxin, and 2 samples (0.6%, 2/357) were not contaminated by any mycotoxin studied. There were 30, 66, and 66 different mycotoxin combination types in CSM, WB, and DDGS, respectively, but DON+3-ADON+15- ADON+ZEA was the most frequent mycotoxin combination, with a frequency of 13.16% (5/38) in CSM, 17.29% (23/133) in WB, and 18.8% (35/186) in DDGS. These results indicate that mycotoxin co-occurrence is very common in Chinese feedstuffs, and it is very important to conduct multi-mycotoxin monitoring in Chinese feedstuffs.