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Mycotoxins contamination is one of the most important problems worldwide in maize that can cause serious threat for human and animal health. The aim of this study was to determine the ability of Fusarium species associated with maize ear rot to produce diverse mycotoxins. The results showed, One out of three isolates of F. subglutinans produced detectable level of Beauvericin (BEA); the only isolate of F. temperatum produced 302 µg/g of BEA. Two out of five isolates of F. redolens produced enniatin B1and four isolates of this species produced high levels of BEA. As well, 21 isolates of Fusarium incarnatum-equiseti species complex (FIESC) and one isolates of F. brachygibbosum were evaluated for production of trichothecens (T-2 toxin, HT-2 toxin, Diacetoxyscirpenol (DAS), nivalenol (NIV) and deoxynivalenol (DON)), zearalenone (ZEN), enniatins (ENN A, A1, B and B1) and BEA by HPLC. Production of trichothecens, ZEN and BEA with one, three and four isolates of FIESC, respectively were observed and one isolate of F. brachygibbosum only produced detectable level of DAS and ENN B1. Analyzing of moniliformin production of F. proliferatum (26 isolates), FIESC (21 isolates) and F.thapsinum (10 isolates) showed none of them produce this toxin. These results revealed the ability of Fusarium spp. from maize to produce a varied range of mycotoxins which are harmful to human and animal’s health. Therefore, the occurrence of such broad number of different species on Iranian maize could be reason of great concern because of the toxigenic risk associated to these species.

Results of mycotoxicological survey of representative samples of feed and procured wheat, barley, oat and maize grain batches are demonstrated. The samples were submitted by the Veterinary Service officials, livestock farmers and feed mill operators, agricultural producers, specialized commercial business operators and farm owners in seven Federal Districts of the Russian Federation in 2009-2019. Similar amounts of wheat and barley grain samples were received from the Central, Volga, Ural and Siberian Federal Districts. The amount of wheat samples delivered from the Southern Federal District prevailed over the number of barley samples, and the maize samples were mostly delivered from the regions of the Central Federal District. Fusarium toxins including T-2 toxin, diacetoxyscirpenol, deoxynivalenol, zearalenone and fumonisins of group B as well as alternariol, ochratoxin A, citrinin, aflatoxin В1, sterigmatocystin, cyclopiazonic acid, mycophenolic acid, ergot alkaloids and emodin were detected and measured according to the validated competitive ELISA procedure. Generalization of the results demonstrated domination of fusarium toxins and active involvement of alternariol in the contamination of all types of feed grains as well as high occurrence of emodin in ear cereals and increased occurrence of T-2 toxin and ochratoxin A in barley. Shift of medians and 90%-percentile of the basic contaminants to lower values as compared to mean and maximal ones was reported thus being indicative of their possible accumulation at the levels outside the typical range. The highest levels of T-2 toxin, deoxynivalenol and ochratoxin A as well as 90%-percentile values exceeded the acceptable levels. The maize grains demonstrated the whole complex of the tested fusarium toxins with the prevalence of T-2 toxin, deoxynivalenol, zearalenone and fumonisins; and the maximal amounts of these mycotoxins by several times exceeded the accepted regulatory levels. Diacetoxyscirpenol, aflatoxin В1, sterigmatocystin, cyclopiazonic acid and ergot alkaloids are classified as rare feed grain contaminants. High prevalence of alternariol and emodin known as “diarrhea factor” as well as maize grain contamination with mycophenolic acid (mycotoxin having an immunosuppressive effect) are for the first time reported in this paper. These data support the need of their introduction in the group of regulated substances significant for public health. Original monitoring data systematized and summarized in the paper are given in electronic format in section Additional materials.Part 1: Veterinary Science Today. 2020; 1 (32): 60–65. DOI: 10.29326/2304-196X-2020-1-32-60-65.

Results of the ten-year annual mycotoxicological testing of complete mixed feeds for pigs and poultry submitted by holdings and processing establishments located in the Northwestern, Central, Southern, Volga and Ural Federal Districts are presented. Competitive ELISA tests showed that the occurrence of T-2 toxin, deoxynivalenol, zearalenon, fumonisines B, alternariol, ochratoxin A, citrinin, mycophenolic acid, ergoalkoloids and emodin was about 5% and quantities thereof varied within one or three orders; quantities of T-2 toxin, deoxynivalenol, zearalenon, fumonisines, and ochratoxin A might exceed maximum admissible levels for feed grains. Diacetoxyscirpenol, aflatoxin В1, sterigmatocystin and cyclopiazonic acid belonged to group of rare contaminants. Level of feed contamination with T-2 toxin and emodin was found to be consistently high during the said period; in some of the years occurrence of deoxynivalenol, fumonisines as well as ochra­toxin A, citrinin, mycophenolic acid and ergoalkoloids increased. In 2016–2018, mixed feed contamination with alternariol increased whereas contamination with fumonizin steadily decreased and level of zearalenon occurrence remained consistently low. Evidence for a wide occurrence of emodin known as “diarrheic factor” as well as for sporadic increase in mixed feed contamination with alternariol, citrinin, mycophenolic acid, mycotoxins having the highly dangerous toxic impact and long-term adverse effects, was detected for the first time. These data confirmed the need for their inclusion into the regulated group of substances significant for public health. General features of pig and poultry feed contamination as well as usefulness of regional surveys for intoxication risk prediction are described. Special attention is paid to the importance of the projects for creation of common information resources that could become a unique scientific basis for innovations in feed poisoning prevention. Original monitoring data systematized and summarized in the paper are given in electronic format in section Additional materials.

Diacetoxyscirpenol (DAS) is a type A trichothecene mycotoxin with low molecular weight, and with respect to its toxicity and the occurrence in food and feed, it is known as a potential risk for public and animal health. In the present study, first, a sensitive and specific monoclonal antibody (5E7) was developed. Then, the antibody was applied to develop a competitive-type pressure-dependent immunosensor (CTPDI). The Au@PtNP was synthesized and labeled with goat antimouse antibody (Au@PtNPs-IgG). Finally, the concentration of DAS was negatively correlated with the pressure signal. In the presence of optimal conditions, matrix-matched calibration curves were plotted for wheat samples, in which an optimal IC50 value (half maximal inhibitory concentration) of 3.08 ng/g was achieved. The CTPDI was further applied to detect natural and blind wheat samples, and validation was carried out by liquid chromatography-tandem mass spectrometry. The results showed that CTPDI was highly appropriate and accurate for detection of DAS in wheat.

Fusarium mycotoxins are common contaminants in cereals and often co-occur with plant-derived mycotoxin sugar conjugates. Several of these modified mycotoxins are not degraded in the small intestine and hence carried through to the large intestine where microbial transformation may occur. This study aims to assess the gastrointestinal stability of the trichothecenes HT-2 toxin (HT-2), HT-2-β-glucoside (HT-2-Glc), diacetoxyscirpenol (DAS), DAS-α-glucoside (DAS-Glc) and fumonisin B1 (FB1), N-(1-deoxy-d-fructos-1-yl) fumonisin-B1 (NDF-FB1). All tested modified mycotoxins were stable under upper gastrointestinal (GI) conditions. In faecal batch culture experiments, HT-2-Glc was hydrolysed efficiently and no further microbial biotransformation of HT-2 was observed. DAS-Glc hydrolysis was slow and DAS was de-acetylated to 15-monoacetoxyscripenol. NDF-FB1 was hydrolysed at the slowest rate and FB1 accumulation varied between donor samples. Our results demonstrate that all tested modified mycotoxins are stable in the upper GI tract and efficiently hydrolysed by human gut microbiota, thus potentially contributing to colonic toxicity. Hence the microbial biotransformation of any novel modified mycotoxins needs to be carefully evaluated.

For maize grain, which has been important for the forage production of the country over the past two decades, persistent contamination with toxins of fusarium fungi, more often T-2/HT-2 toxins, fumonisins, and more rarely with deoxynivalenol and zearalenone in quantities that pose risks for animal intoxication was showed by enzyme immunoassay. The cases of superintensive accumulation of cyclopiazonic acid, citrinin, mycophenolic acid, and ochratoxin A were revealed. In 2016–2018, for maize grain from four subjects of the Central Federal District of the Russian Federation, a significant prevalence of T-2/HT-2 toxins, deoxynivalenol, and fumonisins with fluctuations in the frequency of occurrence of zearalenone over the years was found; diacetoxyscirpenol was detected in several samples from the Kursk and Voronezh oblasts. Contamination of grain with alternariol in 2016 and 2017 was mild both by frequency (5.3%) and accumulation levels (20–85 μg/kg), but the proportion of samples containing this toxin was 40.7% with a content range of 25–295 μg/kg in the yield of 2018. The influence of soil and climatic factors on the nature of mycotoxin contamination of the maize grain yield, the contribution of fungi belonging to dematiaceous hyphomycetes, and the prevailing variants of combined contamination with fusariotoxins are discussed.

For corn grain traded in the country's forage production sector over the past two decades, enzyme immunoassay showed persistent contamination with toxins of fusarium fungi, more often T-2 / HT-2 toxins, fumonisins, and more rarely with deoxynivalenol and zearalenone in quantities that pose risks of animal intoxication. The cases of superintensive accumulation of cyclopiazonic acid, citrinin, mycophenolic acid and ochratoxin A. were revealed. In 2016-2018 For maize grain from 4 subjects of the Central Federal District of the Russian Federation, a significant prevalence of T-2 / HT-2 toxins, deoxynivalenol and fumonisins was found with fluctuations in the frequency of occurrence of zearalenone over the years, diacetoxyscirpenol was detected in several samples from the Kursk and Voronezh regions. Contamination of grain with alternariol in 2016 and 2017 was mild both in frequency (5.3%) and accumulation levels (20–85 µg / kg), but in the collection of 2018, the proportion of samples containing this toxin was 40.7% with a content range of 25 295 µg / kg. The influence of soil and climatic factors on the nature of mycotoxin contamination of corn grain crops, the contribution of fungi belonging to dark-colored hyphomycetes and the prevailing variants of combined contamination with fusariotoxins are discussed.

Introduction and objective Mycotoxins are toxic metabolites produced by certain types of fungi, causing pathological changes in humans and animals. The aim of this study was to assess the degree of contamination of selected cereal grains, bran and cereal products intended for children, with mycotoxins using GCxGC-TOF-MS technique. The study involved mycotoxins belonging to the type A and B trichothecenes group, including T-2 toxin (T-2), HT-2 toxin (HT-2), scirpenol (SCI), 15-monoacetoxyscirpenol (15-MAS), diacetoxyscirpenol (DAS), triacetoxyscirpenol (TAS), fusarenon-X (FUS-X), nivalenol (NIV), deoxynivalenol (DON), 3-acetyl-DON (3-Ac-DON), 15-acetyl-DON (15-Ac-DON). The study also assessed the effect of conditions in which the samples were stored, including temperature (6°C and 28°C) and time (14 and 28 days), on fungal growth and mycotoxin production. Among all studied compounds, only DAS and HT-2 toxins were detected in tested samples, with the exception of products intended for children. Measured HT-2 mycotoxin content in tested samples was in the range 83.9 - 196.4 µg kg -1. Results and conclusions Experiments with storage conditions showed a statistically significant increase in the HT-2 toxin level after 14 days of storage in all samples, irrespective of temperature. Prolonged storage (additional 14 days) did not cause significant changes in the HT-2 content. Further analyses showed a statistically significant effect of storage temperature on HT-2 toxin levels only in cereal products intended for children after both 14 and 28 days. Interestingly, lower temperature (6°C) was more optimal then higher temperature (28°C) for the HT-2 toxin production. No significant effect of storage temperature on HT-2 level was observed for cereal grains and bran.

Background When present in animal feedstuff, mycotoxins contaminants and antibiotic residues can have negative implications for animal production and Public Health, including the transmission of carcinogenic compounds and the selection of antibiotic resistant bacteria, respectively. So far there are no available methods in which both mycotoxins and antibiotic residues are analyzed using a parallel extraction approach. To address this issue, we developed a LC/MS methodology with high sensitivity (0.005 to 6.42 and 24.55 to 132.73 μg kg− 1 for mycotoxins and antimicrobials) and specificity (unique target ion mass/charge) that allows the detection of 26 mycotoxins and 23 antibiotic residues in animal feedstuff and validated it through the determination of these analytes in 294 animal feed and feed ingredient samples in the framework of a country-wide surveillance program. Two hundred and five of these samples were analyzed for mycotoxins and 89 for antibiotics. Findings Fumonisin was the most frequently toxin found, with FB1 and FB2 presenting prevalences of 50 and 52% and maximum concentrations of 14,927.61 and 8646.67 μg kg− 1, respectively. Other toxins, including diacetoxyscirpenol n = 4/101 (3.96%), fusarenon-X n = 2/101 (1.98%), citrinin n = 2 (1.98%), and patulin n = 1 (0.99%) were rarely found. Toxicologically relevant concentrations of toxin metabolites, such as HT-2 (6.38–485.49 μg kg− 1) and 3−/15-acetoxydeoxynivalenol (877.89–3236.56/5.44–1685.3 μg kg− 1), were also found. Few samples exceeded threshold mycotoxin concentrations defined in current EU guidelines. Dairy cattle and swine feeds included the higher number of samples exceeding guideline values (n = 6 and n = 5, respectively). From the total of samples analysed for antibiotics, 7.7% (n = 7/89) were classified as medicated for poultry and pigs. Unexpectedly, 57% of these medicated samples contained no detectable antibiotics (n = 4/7). The remaining 43% of the samples (n = 3/7) presented inconsistencies regarding the concentration of analytes declared on the labels or the antibiotics found. Likewise 74.6% (n = 50/67) of the non-medicated feed samples analyzed had antibiotic residues. Additionally, we analyzed commercial monensin standards for purity and evaluate batch-to-batch flushing feed industry practices. Conclusions Herein we report the results for a year-wide analysis for mycotoxins and antibiotics in feed samples. Mycotoxins, several metabolites, and the occurrence of these emerging contaminants were evaluated and antibiotic residues in non-medicated feed samples were found using a targeted MS-based LC approach. This validated multi-analyte method is expected to facilitate the monitoring and surveillance of contaminants, from natural and anthropogenic origin, in animal feed.

To assess the in vitro activity of three phenolic acids (ferulic acid, p-hydroxybenzoic acid, vanillic acid) and two flavonols (quercetin, rutin) on mycelial growth and mycotoxin accumulation of Fusarium graminearum (FG), F. langsethiae (FL) and F. poae (FP), two different approaches were chosen. First, grains from oat varieties were inoculated with a suspension of three FL isolates to determine the influence of phenolic compounds on the accumulation of mycotoxins. The oat variety Zorro showed a tendency for lower accumulation of T-2/HT-2, diacetoxyscirpenol and neosolaniol. Second, a mycelium growth assay was conducted to follow FG, FL and FP growth on cereal based media supplemented with phenolic compounds. Increasing concentrations of ferulic acid substantially inhibited growth of FG and FL, while FP growth was reduced to 57%. In contrast, p-hydroxybenzoic acid, vanillic acid, quercetin, and rutin slightly stimulated mycelium growth. Results about mycotoxin production in cereal based media were less conclusive.

In this study, a dietary exposure assessment of mycotoxins was conducted for the Romanian population using the contamination data of a various categories of wheat-based products for direct human consumption. Wheat-based foods (n = 181) commercialized in Romania, including flour, bread, biscuits, breakfast cereals and pasta, were evaluated by GC-QqQ-MS/MS for the occurrence of deoxynivalenol (DON), 3-acetyldeoxynivalenol (3AcDON), 15-acetyldeoxynivalenol (15AcDON), fusarenon-X, nivalenol, HT-2 and T-2 toxins, diacetoxyscirpenol, neosolaniol and zearalenone (ZEA). DON and 15AcDON were detected in 63 and 5% of all the analyzed samples, whereas 13-AcDON, HT-2, T-2, NIV and ZEA were not detected. Exposure of Romanian adult population was assessed, the EDIs for the sum of DON+3AcDON+15AcDON were 669 ng kg−1 bw day−1 at low-bound estimation, and 690 ng kg−1 bw day−1 at upper-bound estimation, being lower than the TDI set (1000 ng kg−1 bw day−1).

Mycotoxins have been widely studied by many research groups but further multidisciplinary research is needed to better understand and clarify many issues. This study describes the use of high-performance liquid chromatography coupled with ion trap mass spectrometry (HPLC-MS) to measure T-2 toxin and its metabolites, such as HT-2 toxin, neosolaniol (NEO) and diacetoxyscirpenol (DAS), as well as masked glucosylated mycotoxins in Fusarium-infected Czech spring barley. In total, 152 spring barley samples from the 2018 harvest were analyzed by the ELISA screening method for the presence of T-2 toxin. The most contaminated samples (15), which exceeded the recommended maximum level set by the EU for the sum of T-2 and HT-2 toxin in unprocessed cereals (200 μg/kg), were analyzed by HPLC-MS/MS and microbiological testing. Isolated fungi were evaluated microscopically and identified by polymerase chain reaction (PCR) assays. The prevalence of Fusarium species in spring barley across the Czech Republic in 2018 showed a predominance of F. poae (12 barley samples) and F. tricinctum (9 barley samples). Other strains (F. sporotrichioides and F. langsethiae) were present at a lower frequency, in 1 and 2 samples, respectively. The average concentration of T-2 plus HT-2 toxin was 107.7 μg/kg, while NEO and DAS were found in a few samples at values close to their limit of quantification. HT-2 glucoside was identified in all samples.

A pilot study to investigate the occurrence of 10 mycotoxins (deoxynivalenol, DON, 3-acetyldeoxynivalenol, 3-ADON, 15-acetyldeoxynivalenol, 15-ADON, fusarenon-X, FUS-X, diacetoxyscirpenol, DAS, nivalenol, NIV, neosolaniol, NEO, zearalenone, ZON, zearalanone, ZAN, T-2 toxin, T-2, and HT-2 toxin, HT-2) in esophageal cancer patients was performed with the urinary biomarkers approach in Golestan, Iran. Urine multimycotoxin analysis was performed by dispersive liquid–liquid microextraction and gas chromatography–tandem mass spectrometry (GC–MS/MS) analysis, and values were normalized with urinary creatinine (μg/g). Four mycotoxins, namely NEO (40%), HT-2 (17.6%), DON (10%), and HT-2 (5.8%), were detected in the analyzed urine samples. DON was only detected in the control group (5.09 μg/g creatinine), while T-2 (44.70 μg/g creatinine) was only present in the esophageal cancer group. NEO and HT-2 were quantified in both control and case groups, showing average of positive samples of 9.09 and 10.45 μg/g creatinine for NEO and 16.81 and 29.09 μg/g creatinine for HT-2, respectively. Mycotoxin co-occurrence was observed in three samples as binary (NEO/HT-2 and T-2/HT-2) and ternary (DON/NEO/HT-2) combinations, reaching total concentrations of 44.58, 79.13, and 30.04 µg/g creatinine, respectively. Further investigations are needed to explore a causal association between mycotoxin contamination and esophageal cancer. For this pilot study in Golestan, the low sample size was a very limiting factor.

This study was conducted to investigate human exposure to 19 compounds (mycotoxins and their metabolites) in plasma samples from healthy adults (n = 438, aged 19&ndash, 68 years) from Navarra, a region of northern Spain. Samples were analyzed by LC-MS/MS, before and after enzymatic hydrolysis for the detection of possible glucuronides and/or sulfates (Phase II metabolites). The most prevalent mycotoxin was ochratoxin A (OTA), with an incidence of 97.3%. Positive samples were in the concentration range of 0.4 ng/mL to 45.7 ng/mL. After enzymatic treatment, OTA levels increased in a percentage of individuals, which may indicate the presence of OTA-conjugates. Regarding ochratoxin B, it has also been detected (10% of the samples), and its presence may be related to human metabolism of OTA. Sterigmatocystin was detected with a high incidence (85.8%), but only after enzymatic hydrolysis, supporting glucuronidation as a pathway of its metabolism in humans. None of the other studied mycotoxins (aflatoxins B1, B2, G1, G2 and M1, T-2 and HT-2 toxins, deoxynivalenol, deepoxy-deoxynivalenol, 3-acetyldeoxynivalenol, 15-acetyldeoxynivalenol, zearalenone, nivalenol, fusarenon-X, neosolaniol, and diacetoxyscirpenol) were detected in any of the samples, neither before nor after enzymatic treatment. To the best of our knowledge, this is the first report carried out in Spain to determine the exposure of the population to mycotoxins and some of their metabolites using plasma, and the obtained results justify the need for human biomonitoring and metabolism studies on mycotoxins.

Fusarium incarnatum-equiseti species complex (FIESC) is considered as one of the richest insecticolous species. Fusarium species synthesize toxic secondary metabolites that are not fully understood. Mycotoxin production and pathogenicity on germinating seeds, seedlings, and leaves must be carefully studied for the use of Fusarium species in the biological control of insect pests. In this study, we evaluated the mycotoxin production and phytopathogenic potential of entomopathogenic strains of Fusarium sulawesiensis (1), F. pernambucanum (3), and F. caatingaense (23). The phytopathogenicity tests of F. caatingaense (URM 6776, URM 6777, URM 6778, URM 6779, and URM 6782) were performed during the development of bean (Phaseolus vulgaris, Vigna unguiculata, and Phaseolus lunatus), and corn (Zea mays) seedlings, using four treatments (soil infestation with the inoculum, spraying on leaves, root dip, and negative control). The mycotoxins, monoacetyl-deoxynivalenols (AcDON), deoxynivalenol (DON), beauvericin (BEA), fusarenone-X (FUS), T-2 toxin (T2), diacetoxyscirpenol (DAS), and zearalenone (ZEA), were detected in the study; BEA (detected in 25 strains) and FUS (detected in 21 strains) were found to be predominant. None of the strains showed any ability to cause disease or virulence in beans and corn. The FIESC strains showed a highly variable production of mycotoxins without the potential to be used as phytopathogenic agents for the cultures tested.

Several studies have reported a wide range of severe health effects as well as clinical signs, when livestock animals are exposed to high concentration of mycotoxins. However, little is known regarding health effects of mycotoxins at low levels. Thus, a long-term feeding trial (between May 2017 and December 2019) was used to evaluate the effect of low doses of mycotoxin mixtures on performance of broiler chickens fed a naturally contaminated diet. In total, 18 successive broiler performance trials were carried out during the study period, with approximately 2200 one-day-old Ross-308 chicks used for each trial. Feed samples given to birds were collected at the beginning of each trial and analysed for multi-mycotoxins using a validated LC-MS/MS method. Furthermore, parameters including feed intake, body weight and feed efficiency were recorded on a weekly basis. In total, 24 mycotoxins were detected in samples analysed with deoxynivalenol (DON), zearalenone (ZEN), fumonisins (FBs), apicidin, enniatins (ENNs), emodin and beauvericin (BEV), the most prevalent mycotoxins. Furthermore, significantly higher levels (however below EU guidance values) of DON, ZEN, FBs, BEV, ENNs and diacetoxyscirpenol (DAS) were detected in 6 of the 18 performance trials. A strong positive relationship was observed between broilers feed efficiency and DON (R2 = 0.85), FBs (R2 = 0.53), DAS (R2 = 0.86), ZEN (R2 = 0.92), ENNs (R2 = 0.60) and BEV (R2 = 0.73). Moreover, a three-way interaction regression model revealed that mixtures of ZEN, DON and FBs (p = 0.01, R2 = 0.84) and ZEN, DON and DAS (p = 0.001, R2 = 0.91) had a statistically significant interaction effect on the birds&rsquo, feed efficiency. As farm animals are often exposed to low doses of mycotoxin mixtures (especially fusarium mycotoxins), a cumulative risk assessment in terms of measuring and mitigating against the economic, welfare and health impacts is needed for this group of compounds.

Rye (Secale cereale L.) is one of the most important cereals and is used in both the food and feed industries. It is produced mainly in a belt extending from Russia through Poland to Germany. Despite the great economic importance of this cereal, there is little research on rye contamination with mycotoxins. In this study, the occurrence of Fusarium mycotoxins (deoxynivalenol, nivalenol, 3-acetyl-deoxynivalenol, monoacetoxyscirpenol, diacetoxyscirpenol, T-2 toxin, HT-2 toxin, and zearalenone), as well as ochratoxin A, in 60 winter rye samples of four varieties (KWS Binntto, KWS Serafino, Dańkowskie Granat and Farm Saved Seed) cultivated in three consecutive growing seasons in five different regions of Poland was determined using liquid chromatography with tandem mass spectrometry and fluorescence detection. Deoxynivalenol, T-2 toxin, HT-2 toxin, and zearalenone had the highest occurrence in samples (90%, 63%, 57%, and 45% positive results, respectively). The mean concentrations of these analytes were 28.8 &micro, g/kg (maximum 354.1 &micro, g/kg), 0.98 &micro, g/kg (maximum 6.63 &micro, g/kg), 2.98 &micro, g/kg (maximum 29.8 &micro, g/kg), and 0.69 &micro, g/kg (maximum 10.2 &micro, g/kg), respectively. The mean concentrations for individual mycotoxins were highest in the 2016/2017 growing season. In the 2016/2017 growing season, at least two mycotoxins were detected in 95% of the samples, while in the 2018/2019 growing season, 70% of samples contained one or no mycotoxins. The frequencies of mycotoxin occurrence in different rye varieties were similar. Although a high frequency of mycotoxin occurrence was noted (especially deoxynivalenol), their concentrations were low, and none of the analyzed rye samples exceeded the maximum acceptable mycotoxin level set by the European Commission.

The trichothecene mycotoxins contaminate cereal grains and have been related to alimentary toxicosis resulted in emetic response. This family of mycotoxins comprises type A to D groups of toxic sesquiterpene chemicals. Diacetoxyscirpenol (DAS), one of the most toxic type A trichothecenes, is considered to be a potential risk for human and animal health by the European Food Safety Authority. Other type A trichothecenes, T-2 toxin and HT-2 toxin, as well as type B trichothecene deoxynivalenol (DON), have been previously demonstrated to induce emetic response in the mink, and this response has been associated with the plasma elevation of neurotransmitters peptide YY (PYY) and serotonin (5-hydroxytryptamine, 5-HT). However, it is found that not all the type A and type B trichothecenes have the capacity to induce PYY and 5-HT. It is necessary to identify the roles of these two emetogenic mediators on DAS-induced emesis. The goal of this study was to determine the emetic effect of DAS and relate this effect to PYY and 5-HT, using a mink bioassay. Briefly, minks were fasted one day before experiment and given DAS by intraperitoneally and orally dosing on the experiment day. Then, emetic episodes were calculated and blood collection was employed for PYY and 5-HT test. DAS elicited robust emetic responses that corresponded to upraised PYY and 5-HT. Blocking the neuropeptide Y2 receptor (NPY2R) diminished emesis induction by PYY and DAS. The serotonin 3 receptor (5-HT3R) inhibitor granisetron totally restrained the induction of emesis by serotonin and DAS. In conclusion, our findings demonstrate that PYY and 5-HT have critical roles in DAS-induced emetic response.

Mycotoxins are toxic fungal metabolites, contaminating cereal grains in field or during processing and storage periods. These environmental contaminants pose great threats to humans and animals' health due to their toxic effects. Type A trichothecenes, fumonisins and fusaric acid (FA) are commonly detected mycotoxins produced by various Fusarium species. Trichoderma spp. are promising antagonists in agriculture for their activities against plant pathogens, and also regarded as potential candidates for bioremediation of environmental contaminants. Managing toxigenic fungi by antagonistic Trichoderma is regarded as a sustainable and eco-friendly strategy for mycotoxin control. However, the metabolic activities of Trichoderma on natural occurring mycotoxins were less investigated. Our current work comprehensively explored the activities of Trichoderma against type A trichothecenes, fumonisins and FA producing Fusarium species via co-culture competition and indirect volatile assays. Furthermore, we investigated metabolism of type A trichothecenes and FA in Trichoderma isolates. Results indicated that Trichoderma were capable of bio-transforming T-2 toxin, HT-2 toxin, diacetoxyscirpenol and neosolaniol into their glycosylated forms and one Trichoderma strain could bio transform FA into low toxic fusarinol. These findings proved that Trichoderma isolates could manage toxigenic Fusarium via direct competition and volatile-mediated indirect inhibition. In addition, these antagonists possess defensive systems against mycotoxins for self-protection, which enriches our understanding on the interaction mechanism of Trichoderma spp. on toxigenic fungus.

A mycotoxicological survey was conducted in breakfast (n = 172) and infant (n = 43) cereals commercialized in Brazil. Samples were collected in 2018 for analyses of: aflatoxins (AFs) B1 (AFB1), B2, G1 and G2; fumonisins (FBs) B1 (FB1) and B2; zearalenone (ZEN); the trichothecenes (TRCs) deoxynivalenol (DON), T-2 toxin, HT-2 toxin, nivalenol, fusarenon X, 3-acetyldeoxynivalenol, 15-acetyldeoxynivalenol and diacetoxyscirpenol; and ochratoxin A. FB1 was the most prevalent metabolite in breakfast cereals, being detected in 26.7% of the samples (mean 105 μg/kg); ZEN had the second highest positivity, 14.8% (mean 17 μg/kg), followed by DON with 10% (mean 44 μg/kg). In infant cereals, FB1 also had the highest incidence, 27.8% (mean 55 μg/kg), followed by DON with 10.3% (mean 36 μg/kg) and ZEN with 6.9% (mean 3 μg/kg). Mycotoxins contamination was found in 31.4% (n = 54) of the breakfast cereals and in 18.6% (n = 8) of the infant cereals. In these positive samples, co-occurrence of two or three mycotoxins was detected in 31.5% (n = 17) of the breakfast cereals and in 25% (n = 2) of the infant cereals. The mycotoxins found co-contaminating the breakfast cereals belong to the genera Aspergillus and Fusarium; ZEN, followed by AFB1, were the most prevalent ones. As for the infant cereals, the associated fungal metabolites are produced by the genus Fusarium; the highest incidence was seen for ZEN. Low contamination and positivity of mycotoxins were found herein; nonetheless, in some samples these substances were present at levels which transgress those preconized in the Brazilian legislation. Therefore, mycotoxicological monitoring of this type of product throughout the nation is crucial in order to identify the potential risk to which the Brazilian population is exposed, particularly the children.

Mycotoxins found in randomly selected commercial milk thistle dietary supplement were evaluated for their toxicity in silico and in vitro. Using in silico methods, the basic physicochemical, pharmacological, and toxicological properties of the mycotoxins were predicted using ACD/Percepta. The in vitro cytotoxicity of individual mycotoxins was determined in mouse macrophage (RAW 264.7), human hepatoblastoma (HepG2), and human embryonic kidney (HEK 293T) cells. In addition, we studied the bioavailability potential of mycotoxins and silibinin utilizing an in vitro transwell system with differentiated human colon adenocarcinoma cells (Caco-2) simulating mycotoxin transfer through the intestinal epithelial barrier. The IC50 values for individual mycotoxins in studied cells were in the biologically relevant ranges as follows: 3.57−13.37 nM (T-2 toxin), 5.07−47.44 nM (HT-2 toxin), 3.66−17.74 nM (diacetoxyscirpenol). Furthermore, no acute toxicity was obtained for deoxynivalenol, beauvericin, zearalenone, enniatinENN-A, enniatin-A1, enniatin-B, enniatin-B1, alternariol, alternariol-9-methyl ether, tentoxin, and mycophenolic acid up to the 50 nM concentration. The acute toxicity of these mycotoxins in binary combinations exhibited antagonistic effects in the combinations of T-2 with DON, ENN-A1, or ENN-B, while the rest showed synergistic or additive effects. Silibinin had a significant protective effect against both the cytotoxicity of three mycotoxins (T-2 toxin, HT-2 toxin, DAS) and genotoxicity of AME, AOH, DON, and ENNs on HEK 293T. The bioavailability results confirmed that AME, DAS, ENN-B, TEN, T-2, and silibinin are transported through the epithelial cell layer and further metabolized. The bioavailability of silibinin is very similar to mycotoxins poor penetration.

An analytical method for the simultaneous quantitation of ten trichothecenes of type A (HT-2 toxin, T-2 toxin, diacetoxyscirpenol, and neosolaniol) and type B (3-acetyldeoxynivalenol, 15-acetyldeoxynivalenol, deoxynivalenol, deoxynivalenol-3-glucoside, nivalenol, and fusarenon-X) in feed has been developed using liquid chromatography with tandem mass spectrometry. Mycotoxins extracted twice from samples using aqueous acetonitrile were purified using a multifunctional clean-up column, followed by a phospholipid removal column. Trichothecenes were analysed using liquid chromatography atmospheric pressure chemical ionization tandem mass spectrometry. The extraction efficiency of the mycotoxins and the repeatability of some were improved by repeated extractions. Ionization enhancement (signal enhancement) of some mycotoxins was improved by using the phospholipid removal column at the clean-up step. Spike and recovery tests of trichothecenes were conducted on maize, barley, soybean meal, rapeseed meal, and formula feeds (for starting broiler chicks, suckling pigs, and beef cattle). The mean recovery values were 70.6–119% with relative standard deviations

The occurrence of mycotoxins has become a problem to be discussed, due to its harmfulness to humans and animal’s health, and may be an obstacle to the poultry economy. Mycotoxins are toxic metabolites produced by certain species of fungi and may contaminate food. Aflatoxins are mainly produced by Aspergillus flavus and Aspergillus parasiticus, and B1, B2, G1 and G2 are its best known types. Fumonisin, with its B1, B2 or B3 types, are produced by Fusarium, while ochratoxin A is produced by Penicillium and Aspergillus. The main trichothecenes mycotoxins are T-2 toxin, deoxynivalenol and diacetoxyscirpenol. Zearalenone, produced by different species of Fusarium fungi affects chickens only when they are exposed to extremely high levels of contamination. Generally, immunosuppression, hepatotoxicity and nephrotoxicity as a decrease in performance and production gains are the most observed effects. There are several laboratory methods that can be used for the determination of mycotoxins. In order to control the contamination, it is necessary to adopt proper farming practices which prevent fungi growth. Once grains and feed are contaminated, biological, physical and/or chemical decontamination methods may be employed, although the physical process with adsorbents mixed to the feed is more widely used. Due to the importance of mycotoxins to poultry production, it is necessary to adopt measures to prevent contamination, and also develop a control and an anti-fungal growth and toxin production program by reviewing the critical points favorable to the emergence of toxin-producing fungi.

To date, the use of biomarkers has become generally accepted. Biomarker-driven research has been proposed as a successful method to assess the exposure to xenobiotics by using concentrations of the parent compounds and/or metabolites in biological matrices such as urine or blood. However, the identification and validation of biomarkers of exposure remain a challenge. Recent advances in high-resolution mass spectrometry along with new analytical (post-acquisition data-mining) techniques will improve the quality and output of the biomarker identification process. Chronic or even acute exposure to mycotoxins remains a daily fact, and therefore it is crucial that the mycotoxins' metabolism is unravelled so more knowledge on biomarkers in humans and animals is acquired. This review aims to provide the scientific community with a comprehensive overview of reported in vitro and in vivo mycotoxin metabolism studies in relation to biomarkers of exposure for deoxynivalenol, nivalenol, fusarenon-X, T-2 toxin, diacetoxyscirpenol, ochratoxin A, citrinin, fumonisins, zearalenone, aflatoxins, and sterigmatocystin.

Fusarium Head Blight (FHB) is a destructive disease of small grain cereals and a major food safety concern. Epidemics result in substantial yield losses, reduction in crop quality, and contamination of grains with trichothecenes and other mycotoxins. A number of different fusaria can cause FHB, and there are significant regional differences in the occurrence and prevalence of FHB pathogen species and their associated mycotoxins. Information on FHB pathogen and mycotoxin diversity in Mexico has been extremely limited, but is needed to improve disease and mycotoxin control efforts. To address this, we used a combination of DNA sequence-based methods and in-vitro toxin analyses to characterize FHB isolates collected from symptomatic wheat in Mexico during the 2013 and 2014 growing seasons. Among 116 Fusarium isolates, we identified five species complexes including nine named Fusarium species and 30 isolates representing unnamed or potentially novel species. Significant regional differences (P 0.001) in pathogen composition were observed, with F. boothii accounting for90% of isolates from the Mixteca region in southern Mexico, whereas F. avenaceum and related members of the F. tricinctum species complex (FTSC) accounted for nearly 75% of isolates from the Highlands region in Central Mexico. F. graminearum, which is the dominant FHB pathogen in other parts of North America, was not present among the isolates from Mexico. F. boothii isolates had the 15-acetyldeoxynivalenol toxin type, and some of the minor FHB species produced trichothecenes, such as nivalenol, T-2 toxin and diacetoxyscirpenol. None of the FTSC isolates tested was able to produce trichothecenes, but many produced chlamydosporol and enniatin B.

The awareness of mycotoxin-related issues in aquaculture industry has grown as manufacturers and producers realise the importance of mycotoxins, beyond aflatoxin, and their potential to impact production. During a period of one year (January 2016 – December 2016), 175 samples of different plant proteins, aquaculture/fishery by-products and finished aquaculture feeds, were analysed. Samples were tested for aflatoxins (aflatoxin B1, B2, G1 and G2); zearalenone; type B trichothecenes (deoxynivalenol (DON); nivalenol; 3-acetyldeoxynivalenol; 15-acetyldeoxynivalenol and fusarenon X-glucoside); fumonisins (fumonisin B1, B2 and B3); type A trichothecenes (T-2 and HT-2 toxin; diacetoxyscirpenol and neosolaniol); and ochratoxin A. All samples were sourced in Southeast Asia. Within the analysed samples, 4% were free of detectable mycotoxins. 8% of the samples had one mycotoxin and 88% of the samples were contaminated with more than one mycotoxin. The current work show similarity to previous studies, that mycotoxin occurrence in plant-meals, and consequently in finished feeds, is quite variable and confirms that aflatoxin is not the main mycotoxin in aquafeeds. Mycotoxin occurrence in fish and shrimp feeds in the present samples were higher than reported in previous studies for the same region. In samples of shrimp feed with DON detected, the mean average DON level was 882 μg/kg with a maximum level of 2,287 μg/kg. These values are within the reported sensitivity levels of white leg shrimp (Litopenaeus vannamei). Despite the limited number and short period of sampling, this inventory study helps address a knowledge gap of mycotoxin occurrence in feeds specifically destined for the aquaculture industry.

Fusarium mycotoxins were determined by a multi-mycotoxin liquid chromatography-tandem mass spectrometry (LC-MS/MS) method in cassava ( garri and lafun ) and yam ( amala ) products collected from some selected Nigerian markets. Of the 94 samples analysed, 71% were contaminated with at least one mycotoxin at an incidence rate of 54% ( garri ), 72% ( lafun ), and 82% ( amala ). Deoxynivalenol which ranged between 35 and 99 μg/kg was the most prevalent Fusarium mycotoxin in garri with an incidence rate of 38%, while the dominant mycotoxin in lafun (61%, range = 30–392 μg/kg) and amala (68%, range = 29–155 μg/kg) was fumonisin B 2 . Other mycotoxins detected from the food products included fumonisin B 1 and B 3 , zearalenone, 3-acetyl-deoxynivalenol, 15-acetyl-deoxynivalenol, deoxynivalenol-3-glucoside, T-2 toxin, diacetoxyscirpenol, and fusarenon-X. The consumption of these contaminated products may be a possible route of exposure to a cocktail of Fusarium mycotoxins and can cause potential synergistic health effects on the Nigerian population.

We surveyed the presence of 22 mycotoxins in 191 Spanish cow milk samples. Mycotoxins could be carried over from diet into animal milk and have toxic effects on human and animal health. The interaction of different mycotoxins may be additive or synergetic. Therefore, surveillance of mycotoxin co-occurrence in milk is recommended. Aflatoxins M1, B1, B2, G1, and G2, ochratoxins A and B, nivalenol, deoxynivalenol, deepoxy-deoxynivalenol, 3- and 15-acetyldeoxynivalenol, diacetoxyscirpenol, neosolaniol, fusarenon X, T-2 and HT-2 toxins, fumonisins B1, B2, and B3, sterigmatocystin, and zearalenone were analyzed. Samples were treated by liquid-liquid extraction with acidified acetonitrile, followed by an acetonitrile-water phase separation using sodium acetate. The analysis was carried out by HPLC coupled to a triple quadrupole mass spectrometer. None of the analyzed mycotoxins had a concentration level higher than their detection limit (0.05-10.1 µg/L). The aflatoxin M1 in the samples never exceeded the level established by the European Union.

Exposure to environmental contaminants might play an important role in neurodegenerative disease pathogenesis, such as Parkinson´s disease (PD) and Alzheimer´s disease (AD). For the first time in Spain, the plasmatic levels of 19 mycotoxins from patients diagnosed with a neurodegenerative disease (44 PD and 24 AD) and from their healthy companions (25) from La Rioja region were analyzed. The studied mycotoxins were aflatoxins B1, B2, G1, G2 and M1, T-2 and HT-2, ochratoxins A (OTA) and B (OTB), zearalenone, sterigmatocystin (STER), nivalenol, deoxynivalenol, 3-acetyldeoxynivalenol, 15-acetyldeoxynivalenol, deepoxy-deoxynivalenol, neosolaniol, diacetoxyscirpenol and fusarenon-X. Samples were analyzed by LC-MS/MS before and after treatment with β-glucuronidase/arylsulfatase in order to detect potential metabolites. Only OTA, OTB and STER were detected in the samples. OTA was present before (77% of the samples) and after (89%) the enzymatic treatment, while OTB was only detectable before (13%). Statistically significant differences in OTA between healthy companions and patients were observed but the observed differences might seem more related to gender (OTA levels higher in men, p-value = 0.0014) than the disease itself. STER appeared only after enzymatic treatment (88%). Statistical analysis on STER, showed distributions always different between healthy controls and patients (patients’ group &gt, controls, p-value &lt, 0.0001). Surprisingly, STER levels weakly correlated positively with age in women (rho = 0.3384), while OTA correlation showed a decrease of levels with age especially in the men with PD (rho = −0.4643).

Our research was focused to identify the Botrytis, Fusarium and Rhizopus species from grapes of the Slovak origin. A further goal of the project was to characterized toxinogenic potential of chosen strains of species Fusarium. 50 samples of grapes, harvested in years 2011, 2012 and 2013 from various wine-growing regions were analyzed in this study. For the isolation of species the of direct plating method was used: a) surface-sterilized berries (using 1% freshly pre-pared chlorine) b) berries and c) damaged berries on DRBC (Dichloran Rose Bengal Chloramphenicol agar). For each analysis were used 50 berries (or all damaged berries from sample). The cultivation was carried at 25 ±1°C, for 5 to 7 days in dark. After incubation, the colonies of Botrytis, Fusarium and Rhizopus were transferred to identification media and after incubation strains were identified to species level. Thirteen species of fusaria (F. acuminatum, F. avenaceum, F. culmorum, F. equiseti, F. graminearum, F. oxysporum, F. proliferatum, F. semitectum, F. solani, F. sporotrichioides,F. subglutinans F. tricinctum and F. verticilioides) were identified. Frequency of fusaria isolation was 92 %. Botrytis cinerea was determined from 86% samples and Rhizopus from 94%. Chosen strains of species of genus Fusarium were able to produce following mycotoxins: deoxynivalenol, T-2 toxin, HT-2 toxin and diacetoxyscirpenol in in vitro conditions as determinated by thin-layer chromatography. Thirty-two (68%) of tested isolates of Fusarium species were able to produce at least one mycotoxin.

Maize is the staple food of Zimbabweans and is consumed daily in the majority of households, particularly subsistence farming households. The objectives of this study were first, to determine the occurrence of mycotoxins in maize produced and consumed by subsistence farmers in Zimbabwe and second, to determine mycotoxin exposure through maize consumption and subsequently the human health risk. A total of 95 maize meal samples were collected from the household stores of randomly selected subsistence farming households. Maize intake data and agronomic practices of these households were investigated A multi-mycotoxin LC-MS/MS method was used to analyze and quantify mycotoxin contamination in the maize samples. Mycotoxin contamination was compared across agro-ecological zones in order to determine differences in mycotoxin contamination levels and presented. Of the toxicologically relevant mycotoxins, aflatoxin B1 (AFB1), fumonisin B1 (FB1), FB2, deoxynivalenol (DON) and zearalenone (ZEN) were detected in 1, 95, 31, 24 and 15 % of the samples at mean levels of 11, 242, 120, 217 and 110 μg/kg respectively. Other mycotoxins detected in the maize were 15-acetyl-deoxynivalenol (15-ADON), nivalenol (NIV), FB3, alternariol-methylether (AME), AFB2, AFG1 and diacetoxyscirpenol (DAS) and the percentage contamination ranged between 1 and 4 % in the maize samples. Contamination of the maize by the mycotoxins was observed at minimum levels below limit of detection for each mycotoxin and maximum levels of 105, 530, 67, 108, 3, 4 and 14 μg/kg for 15-ADON, NIV, FB3, AME, AFB2, AFG1 and DAS respectively. The median levels of each mycotoxin were reported below the limit of detection, with the exception of FB1 (median, 146 μg/kg), which was further considered in the exposure and risk assessment. Dietary exposure was derived from combining mean maize intake data and median FB1 contamination. Mean maize intake was estimated to be 26.8, 37.2, 30.1, 15.8 and 15.0 g/kg bodyweight (bw)/day for under 5s, children, adolescents, adults and the elderly respectively. Subsequently FB1 exposure from maize was calculated, to be 3.91, 5.40, 4.40, 2.30 and 2.20 μg/kg bw/day for these populations. Exposure to FB1 through maize intake was observed to equate to 196, 272, 220, 115 and 110 % of the provisional maximum tolerable daily intake (2 μg/kg bw/day) for under 5s, children, adolescents, adults and the elderly respectively. Subsistence farming communities in Zimbabwe are at risk of high exposure to FB1 and the risk was highest for under 5s, children and adolescents respectively.

Several studies reported adverse respiratory health effects in workers exposed to ambient contaminants in bakeries. The aim of this study was to examine worker exposure to fungi and mycotoxins in Portuguese bakeries in order to develop new policies in occupational health. Environmental samples such as air, surfaces, settled dust and electrostatic dust collector (EDC) were collected in 13 bakeries for fungal and mycotoxins assessment. Air samples obtained by impaction were performed applying malt extract agar (MEA) supplemented with chloramphenicol (0.05%) and dichloran glycerol (DG18) agar-based media. Air samples collected through impinger method were determined as well for fungal detection by molecular tools of Aspergillus sections and mycotoxins. The highest median value for fungal load was 1053 CFU&middot, m&minus, 3 and 65.3% (32 out of 49) of the sampling sites displayed higher fungal load than limits imposed by the World Health Organization. Aspergillus genera was found in air, surface swabs and EDC. Molecular tools were effective in measuring Aspergillus section Fumigati in 22.4% on air, 27.8% on surface swabs and in 7.4% in EDC and Aspergillus section Versicolores in one air sample. All settled dust samples showed contamination with six to eight mycotoxins in each sample. The mycotoxins detected were deoxynivalenol-3-glucoside, deoxynivalenol, zearalenone, 15-acetyldeoxynivalenol, monoacetoxyscirpenol, diacetoxyscirpenol, fumonisin B1, fumonisin B2, griseofulvin, HT2, ochratoxin A, ochratoxin B and mycophenolic acid. Industrial hygienists and exposure assessors should rely on different sampling methods (active and passive) and different assays (culture based and molecular methods) to obtain an accurate risk characterization regarding fungal burden (fungi and mycotoxins). Additionally, the awareness for the raw material as a potential mycotoxins indoor contamination source is important.

Twelve species of bacteria and two species of yeast were tested for sensitivity against 11 different mycotoxins using a disc diffusion assay. Among the bacterial species, Bacillus brevis appeared to be the most sensitive microorganism, being sensitive to eight mycotoxins (AFB1, ochratoxin A [OA], citrinin [CT], patulin [PAT], penicillic acid [PA], cyclopiazonic acid [CPA], penitrem A [PT-A], and zearalenone [ZEE]). This microorganism was not affected by a high concentration (500 μg per disc) of any of the trichothecenes (T-2 toxin, HT-2 toxin, diacetoxyscirpenol, and deoxynivalenol). Kluyveromyces marxianus , a species of yeast, was the only microorganism that was inhibited by all four of the trichothecenes but was not inhibited by the other mycotoxins. The area of the inhibition zone produced by some of the mycotoxins such as OA with the B. brevis assay was dramatically influenced by the pH of the medium, while the toxicity of other mycotoxins such as AFB1 was relatively pH independent. The sensitivity of the B. brevis assay also tended to decrease at agar volumes above 6 ml and as the number of microbes per plate increased. The lowest amounts of the different ochratoxins; OA, OB and OC (OA ethyl ester) that could be detected under optimal conditions were 0.5, 20, and 2 μg per disc, respectively. The lowest amounts of CPA, AFB1 CT, PAT, PA, PT-A, and ZEE that were detectable were 0.5, 1, 1, 1, 1, 1, and 10 μg per disc, respectively. These results demonstrate that B. brevis can be used as a positive indicator organism to detect the presence of several common non-trichothecene mycotoxins. The results demonstrate that, used together, B. brevis and K. marxianus can be used as indicator organisms in a bioassay approach to the detection of several of the most common mycotoxins.

To evaluate the influence of weather conditions on mycotoxin presence in wheat, deoxynivalenol (DON), 3-acetyldeoxynivalenol (3AcDON), 15-acetyldeoxynivalenol (15AcDON), fusarenon-X (FUS-X), nivalenol (NIV), HT-2 toxin (HT-2), T-2 toxin (T-2), diacetoxyscirpenol (DAS), neosolaniol (NEO) and zearalenone (ZEN) were evaluated in 102 Romanian wheat samples coming from five wheat growing areas during 2015. Only six mycotoxins were detected, while FUS-X, DAS, NEO and NIV were not present in the wheat samples. Mycotoxin concentrations were correlated with precipitation and temperature values during anthesis and the preharvest period. Overall, the highest frequency was registered for DON, while the lowest frequency was registered for NIV. In the North Muntenia, DON and ZEN registered high frequencies (68% and 16%, respectively). This region was characterized in June and July by medium to high values of rainfall (41&ndash, 100 mm/month) and normal temperatures (mean of 20.0 &deg, C in June and 24.0 &deg, C in July), suggesting that precipitation levels influence fungi and mycotoxin development to a greater extent compared to the influence of temperature.

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), &alpha, zearalenol (&alpha, ZAL), &beta, zearalenol (&beta, 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 &alpha, 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 &mu, g/kg (regulatory level in diets for lactating dairy animals) and HT-2 content in five barley (7%) and four wheat (5%) samples exceeded 100 &mu, g/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.

Data from many parts of the world on incidence and levels of aflatoxins in feeds and feed ingredients indicate cause for concern, especially in tropical and sub-tropical regions and in those countries importing feedstuffs from these areas. The main products affected are peanut (groundnut) meal, cottonseed products, corn, copra products, and mixed feeds containing any of these components. Reports of the presence of the metabolite aflatoxin M1 in dairy products provide further evidence of contaminated feeds. Although the main survey effort has been with the aflatoxins, surveys in several countries for other fungal toxins have revealed some contamination of certain grains or pasture grasses with ochratoxin A, citrinin, sterigmatocystin, patulin, penicillic acid, zearalenone, trichothecenes (T-2 toxin, deoxynivalenol, nivalenol, and diacetoxyscirpenol), alternariol and its methyl ether, PR toxin, kojic acid, ergot alkaloids, and sporidesmin. It is important to differentiate between biased and unbiased surveys in evaluation of these data. Mycotoxins usually result from improper storage of feedstuffs, although in certain cases they can also be formed before harvest. Tenuazonic acid has been detected in blast diseased rice plants. Dicoumarol may be produced in sweet clover as a result of fungal activity. Ipomeamarone and related compounds in the sweet potato are examples of toxins synthesized by plants in response to invasion by molds or to other stress conditions.

In recent years, the less-studied Alternaria mycotoxins have attracted increasing interest due to the lack of survey data and their ability to cause toxic effects in animals and humans. To fill the gap, the aim of this three-year survey was to investigate the presence and co-occurrence of Alternaria and other mycotoxins in a total of 433 cereal grain samples from Slovenian farms and agricultural cooperatives from 2014 to 2016. Using the multi-mycotoxin method, 14 mycotoxins were determined. In 53% of 433 analysed samples, contamination with at least one mycotoxin was found. Deoxynivalenol (DON) and tenuazonic acid (TeA) were present in 32% and 26% of cereal grain samples, respectively, whereas alternariol (AOH), tentoxin (TEN), alternariol monomethyl ether (AME), 3- and 15-acetyldeoxynivalenol (3- and 15-AcDON), and zearalenone (ZEN) were present in fewer than 15% of the samples. Ochratoxin A (OTA) was found in one rye sample, while diacetoxyscirpenol (DAS), HT-2 and T-2 toxin, and fumonisins B1 and B2 (FB1 and FB2) were not detected. The highest maximum and median concentrations of Alternaria toxins were determined in spelt in 2016 (TeA, 2277 µg/kg and 203 µg/kg, respectively), and those of Fusarium toxins in wheat in 2015 (DON, 4082 µg/kg and 387 µg/kg, respectively). The co-occurrence of two or more mycotoxins was found in 43% of the positive samples. The correlations between Alternaria toxins were very weak but statistically significant (r: 0.15–0.17, p: 0.0042–0.0165). A well-known correlation between Fusarium toxins DON and ZEN was weak and highly significant (r = 0.28, p &lt, 0.0001).

Multiple mycotoxins were tested in milled rice samples (n = 200) from traders at different milling points within the Mwea Irrigation Scheme in Kenya. Traders provided the names of the cultivar, village where paddy was cultivated, sampling locality, miller, and month of paddy harvest between 2018 and 2019. Aflatoxin, citrinin, fumonisin, ochratoxin A, diacetoxyscirpenol, T2, HT2, and sterigmatocystin were analyzed using ultra-high-performance liquid chromatography–tandem mass spectrometry (UHPLC–MS/MS). Deoxynivalenol was tested using enzyme-linked immunosorbent assay (ELISA). Mycotoxins occurred in ranges and frequencies in the following order: sterigmatocystin (0–7 ppb, 74.5%), aflatoxin (0–993 ppb, 55.5%), citrinin (0–9 ppb, 55.5%), ochratoxin A (0–110 ppb, 30%), fumonisin (0–76 ppb, 26%), diacetoxyscirpenol (0–24 ppb, 20.5%), and combined HT2 + T2 (0–62 ppb, 14.5%), and deoxynivalenol was detected in only one sample at 510 ppb. Overall, low amounts of toxins were observed in rice with a low frequency of samples above the regulatory limits for aflatoxin, 13.5%, ochratoxin A, 6%, and HT2 + T2, 0.5%. The maximum co-contamination was for 3.5% samples with six toxins in different combinations. The rice cultivar, paddy environment, time of harvest, and millers influenced the occurrence of different mycotoxins. There is a need to establish integrated approaches for the mitigation of mycotoxin accumulation in the Kenyan rice.

Trichothecene mycotoxins are sesquiterpenoid compounds primarily produced by fungi in taxonomical genera such as Fusarium, Myrothecium, Stachybotrys, Trichothecium, and others, under specific climatic conditions on a worldwide basis. Fusarium mold is a major plant pathogen and produces a number of trichothecene mycotoxins including deoxynivalenol (or vomitoxin), nivalenol, diacetoxyscirpenol, and T-2 toxin, HT-2 toxin. Monogastrics are sensitive to vomitoxin, while poultry and ruminants appear to be less sensitive to some trichothecenes through microbial metabolism of trichothecenes in the gastrointestinal tract. Trichothecene mycotoxins occur worldwide however both total concentrations and the particular mix of toxins present vary with environmental conditions. Proper agricultural practices such as avoiding late harvests, removing overwintered stubble from fields, and avoiding a corn/wheat rotation that favors Fusarium growth in residue can reduce trichothecene contamination of grains. Due to the vague nature of toxic effects attributed to low concentrations of trichothecenes, a solid link between low level exposure and a specific trichothecene is difficult to establish. Multiple factors, such as nutrition, management, and environmental conditions impact animal health and need to be evaluated with the knowledge of the mycotoxin and concentrations known to cause adverse health effects. Future research evaluating the impact of low-level exposure on livestock may clarify the potential impact on immunity. Trichothecenes are rapidly excreted from animals, and residues in edible tissues, milk, or eggs are likely negligible. In chronic exposures to trichothecenes, once the contaminated feed is removed and exposure stopped, animals generally have an excellent prognosis for recovery. This review shows the occurrence of trichothecenes in food and feed in 2011–2020 and their toxic effects and provides a summary of the discussions on the potential public health concerns specifically related to trichothecenes residues in foods associated with the exposure of farm animals to mycotoxin-contaminated feeds and impact to human health. Moreover, the article discusses the methods of their detection.

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 for all the analytes. The method accuracy was evaluated by recovery assays at three concentration levels, 25, 50 and 100 μg kg(-1) (n = 9). Recoveries ranged from 62% to 97% and precision, expressed as intra- and inter-day relative standard deviations, was evaluated for all compounds. The validated method was successfully applied to the analysis of 35 laboratory rat feed samples showing mycotoxin contamination in 66% of the samples. DON was the most prevalent trichothecene followed by 15-ADON, NIV and 3-ADON. The maximum DON concentration reached in real samples was 2156 ± 4.3 μg kg(-1), while NEO, DAS and FUS-X were not detected in any sample. Multi-contamination by at least two mycotoxins was observed in 17% of the analysed feed samples.

An LC-MS/MS (QqQ) method has been developed and validated for simultaneous determination of the following trichothecenes in UHT cow milk: nivalenol (NIV), deoxynivalenol (DON), deepoxy-deoxynivalenol (DOM-1), 3-acetyldeoxynivalenol (3-ADON), 15-acetyldeoxynivalenol (15-ADON), neosolaniol (NEO), diacetoxyscirpenol (DAS), fusarenon X (FUS-X), T-2 and HT-2 toxins. Sample treatment is simple and based on the extraction with acetonitrile (ACN), acidified with 0.2% formic acid, followed by a purification process, adding sodium acetate to the ACN/water extract in order to separate aqueous phase and, consequently, polar components of the milk. Validation of the method for all the 10 mycotoxins was successful; validation parameters taken into account were as follows: limits of detection (LOD) and quantification (LOQ), linearity, precision (within-day and between-day variability), recovery, matrix effect and stability. The LODs were 10.1, 2.5, 1.5, 1.9, 0.1, 0.5, 1.0, 0.08, 0.4 and 0.05ng/mL for NIV, DON, DOM-1, FUS-X, NEO, 3-ADON, 15-ADON, DAS, HT-2 and T-2, respectively. Mean recovery values (obtained in intermediate precision conditions) were between 63.5 and 75.8 (RSDR≤15%) for all the mycotoxins. All the mycotoxins suffered from matrix effects, especially DON.

Mycotoxins are secondary metabolites produced by filamentous fungi that usually contaminate food products. Coffee is a natural product susceptible to mycotoxin contamination. The present study evaluates the presence of nivalenol, deoxynivalenol, T-2 and HT-2 Toxin, diacetoxyscirpenol, aflatoxin B 1 , aflatoxin B 2 , aflatoxin G 1 , aflatoxin G 2 , fumonisin B 1 , fumonisin B 2 , ochratoxin A, zearalenone, enniatin A, enniatin A 1 , enniatin B, enniatin B 1 , and beauvericin in coffee samples, using liquid chromatography tandem mass spectrometry (LC-MS/MS). The results show that zearalenone was not present in any sample. In the positive samples the contents of fumonisins ranged from 58.62 to 537.45 μg/kg, emerging mycotoxins ranged from 0.10 to 3569.92 μg/kg, aflatoxins ranged from 0.25 to 13.12 μg/kg, and trichothecenes, excepting nivalenol, ranged from 5.70 to 325.68 μg/kg. Nivalenol presented the highest concentrations, from 0.40 to 25.86 mg/kg. Ochratoxin A ranged from 1.56 to 32.40 μg/kg, and five samples exceeded the maximum limit established by the European Commission.

A multi-mycotoxin chromatographic method was developed and validated for the simultaneous quantitation of aflatoxins (AFB1, AFB2, AFG1 and AFG2), ochratoxin A (OTA), zearalenone (ZON), deoxynivalenol (DON), nivalenol (NIV), diacetoxyscirpenol (DAS), fumonisins (FB1, FB2 and FB3), T-2 toxin (T-2) and HT-2 toxin (HT-2) in feed. The three most popular sample preparation techniques for determination of mycotoxins have been evaluated, and the method with highest recoveries was selected and optimized. This modified QuEChERS (quick, easy, cheap, effective, rugged and safe) approach was based on the extraction with acetonitrile, salting-out and cleanup with lipid removal. A reconstitution process in methanol/water was used to improve the MS responses and then the extracts were analyzed by LC-MS/MS. In this method, the recovery range is 70&ndash, 100% for DON, DAS, FB1, FB2, FB3, HT-2, T-2, OTA, ZON, AFG1, AFG2, AFB1 and AFB2 and 55% for NIV in the spike range of 2&ndash, 80 &micro, g/kg. Method robustness was determined with acceptable z-scores in proficiency tests and validation experiments.

We report the methodology for the quantification of 19 mycotoxins in human plasma using high performance liquid chromatography-mass spectrometry (triple quadrupole). The studied mycotoxins were: deepoxy-deoxynivalenol, aflatoxins (B1, B2, G1, G2 and M1), T-2 and HT-2, ochratoxins A and B, zearalenone, sterigmatocystin, nivalenol, deoxynivalenol, 3-acetyldeoxynivalenol, 15-acetyldeoxynivalenol, neosolaniol, diacetoxyscirpenol and fusarenon-X. Sample deproteinization and cleanup were performed in one step using Captiva EMR-lipid (3 mL) cartridges and acetonitrile (with 1% formic acid). The extraction step was simple and fast. Validation was based on the evaluation of limits of detection (LOD) and quantification, linearity, precision, recovery, matrix effect, and stability. LOD values ranged from 0.04 ng/mL for aflatoxin B1 to 2.7 ng/mL for HT-2, except for nivalenol, which was 9.1 ng/mL. Recovery was obtained in intermediate precision conditions and at three concentration levels. Mean values ranged from 68.8% for sterigmatocystin to 97.6% for diacetoxyscirpenol (RDS ≤ 15% for all the mycotoxins). Matrix effects (assessed at three concentration levels and in intermediate conditions) were not significant for most of the mycotoxins and were between 75.4% for sterigmatocystin and 109.3% for ochratoxin B (RDS ≤ 15% for all the mycotoxins). This methodology will be useful in human biomonitoring studies of mycotoxins for its reliability.

International audience; 4,15-Diacetoxyscirpenol (DAS) is a mycotoxin primarily produced by Fusarium fungi and occurring predominantly in cereal grains. As requested by the European Commission, the EFSA Panel on Contaminants in the Food Chain (CONTAM) assessed the risk of DAS to human and animal health related to its presence in food and feed. Very limited information was available on toxicity and on toxicokinetics in experimental and farm animals. Due to the limitations in the available data set, human acute and chronic health-based guidance values (HBGV) were established based on data obtained in clinical trials of DAS as an anticancer agent (anguidine) after intravenous administration to cancer patients. The CONTAM Panel considered these data as informative for the hazard characterisation of DAS after oral exposure. The main adverse effects after acute and repeated exposure were emesis, with a no-observed-adverse-effect level (NOAEL) of 32 lg DAS/kg body weight (bw), and haematotoxicity, with a NOAEL of 65 lg DAS/kg bw, respectively. An acute reference dose (ARfD) of 3.2 lg DAS/kg bw and a tolerable daily intake (TDI) of 0.65 lg DAS/kg bw were established. Based on over 15,000 occurrence data, the highest acute and chronic dietary exposures were estimated to be 0.8 and 0.49 lg DAS/kg bw per day, respectively, and were not of health concern for humans. The limited information for poultry, pigs and dogs indicated a low risk for these animals at the estimated DAS exposure levels under current feeding practices, with the possible exception of fattening chicken. Assuming similar or lower sensitivity than for poultry, the risk was considered overall low for other farm and companion animal species for which no toxicity data were available. In consideration of the similarities of several trichothecenes and the likelihood of co-exposure via food and feed, it could be appropriate to perform a cumulative risk assessment for this group of substances.

Simultaneous determination of twenty-seven mycotoxins in ready-to-eat food samples using &ldquo, Quick Easy Cheap Rough and Safe&rdquo, (QuEChERS) extraction and chromatographic methods coupled to mass spectrometry in tandem is described in this study. Mycotoxins included in this survey were aflatoxins (B1, B2, G1, G2), enniatins (A, A1, B, B1), beauvericin (BEA), fumonisins (FB1, FB2), sterigmatocystin (STG), deoxynivalenol (DON), 3-acetyl-deoxynivalenol (3-ADON), 15-acetyl-deoxynivalenol (15-ADON), nivalenol (NIV), neosolaniol (NEO), diacetoxyscirpenol (DAS), fusarenon-X (FUS-X), zearalenone (ZEA), &alpha, zearalanol (&alpha, ZAL), &beta, zearalenone (&beta, ZAL), &alpha, zearalenol (&alpha, ZOL), &beta, zearalenol (&beta, zol), T2, and HT-2 toxin. The method showed satisfactory extraction results with recoveries ranging from 63 to 119% for the different food matrix samples. Limits of detection (LODS) and quantification (LOQs) were between 0.15&ndash, 1.5 &micro, g/kg and 0.5&ndash, 5 &micro, g/kg, respectively. The method was successfully applied to the analysis of 25 ready-to-eat food samples. Results showed presence of deoxynivalenol at 36% of samples (2.61&ndash, 21.59 &micro, g/kg), enniatin B at 20% of samples (9.83&ndash, 86.32 &micro, g/kg), HT-2 toxin at 16% of samples (9.06&ndash, 34.43 &micro, g/kg), and aflatoxin G2 at 4% of samples (2.84 &micro, g/kg). Mycotoxins were detected mainly in ready-to-eat food samples prepared with cereals, vegetables, and legumes, even at levels below those often obtained from raw food.

4,15-Diacetoxyscirpenol (4,15-DAS) is a type A trichothecene mycotoxin produced by Fusarium species. Four modified forms of 4,15-DAS including 7-hydroxydiacetoxyscirpenol, 7,8-dihydroxydiacetoxyscirpenol, 4β,8α,15-triacetoxy-3α,7α-dihydroxy-12,13-epoxytrichothec-9-ene and 4,15-diacetylnivalenol were purified from cultures of F. equiseti. An analytical method using a multifunctional column has been developed for the simultaneous determination of 4,15-DAS, its four modified forms, T-2 toxin, HT-2 toxin and neosolaniol in cereals. The performance of the current method was evaluated, and a total of 248 samples of five different commodities were analyzed for over two years by this method. 4,15-DAS was detected in Job’s tears products, corn flour and azuki bean, but it was not found in wheat flour or rye flour. The four modified forms of 4,15-DAS were detected in samples of Job’s tears products, contaminated by 4,15-DAS. This is the first report on quantification of the modified forms of 4,15-DAS in cereals.