Deoxynivalenol and zearalenone in Brazilian barley destined for brewing.

Barley is an important cereal worldwide. However, fungal contamination during pre and postharvest is a recurrent problem for barley production, causing a direct impact on the quality of the grains and their by-products due to spoilage and mycotoxin accumulation. The Fusarium graminearum species complex is the main contaminant during preharvest and some species can produce deoxynivalenol and zearalenone, important mycotoxins that represent a risk to human and animal health. This study evaluated the fungal diversity and the levels of DON and ZEN in barley grains produced in Brazil. The results showed high frequency (60%) of Fusariumcontamination in barley grains. Additionally, mycotoxin levels ranged from 46 to 2074 µg/kg for DON and from 74 to 556 µg/kg for ZEN. Co-occurrence of DON and ZEN was observed in 40% of the samples and 30% of barley samples had DON and ZEN levels higher than the maximum levels established by Brazilian and European legislations. .

Rapid assessment of enniatins in barley grains using near infrared spectroscopy and chemometric tools.

Barley is an important crop worldwide, and it can be affected by various fungi, among them Fusarium is one of the most relevant due to the economic losses caused by mycotoxin contamination. Enniantins (ENNs) are one of the emergent group of mycotoxins that have been found in grains around the world. Nowadays, the main analytical tools available to evaluate these contaminants are based on chromatographic techniques that are efficient but time-consuming and expensive. In this context, the present study aimed to assess the performance of near infrared (NIR) spectroscopy to detect and/or classify the enniatin (ENN) content on barley grains. Sixty samples of barley grains from three different regions of Brazil were investigated and the ENN content determined by UPLC-MS/MS. The levels found were then used to develop multivariate models based on infrared spectral data. The results indicated high incidence off ENN presence in the samples (>70 %) and the PLS-DA model determined by NIR data showed adequate values of sensitivity and sensibility (100 % and 94.2 %, respectively) distinguishing between contaminated and non-contaminated barley samples, demonstrating NIR as a promising tool to monitoring this emergent mycotoxin.

Characterization of the Fusarium sambucinum species complex and detection of multiple mycotoxins in Brazilian barley samples.

This study investigated the fungal diversity in Brazilian barley samples, focusing on the Fusarium sambucinum species complex and the presence of multiple mycotoxins: aflatoxins B1, B2, G1, G2 beauvericin (BEA), enniatins (ENNs) A, A1, B, and B1, deoxynivalenol (DON), fumonisins (FB) B1 and B2, HT-2 and T-2 toxins, nivalenol (NIV) and ochratoxin A (OTA) from two different regions, São Paulo (SP) and Rio Grande do Sul (RS). The majority of the isolates belonged to the Fusarium sambucinum species complex (FSAMSC), with F. graminearum s.s. characterized as the major contaminant. F. meridionale and F. poae were the second most frequent fungi isolated from SP and RS, respectively. All of the F. graminearum s.s. isolates demonstrated 15-ADON genotype, whereas F. poae and F. meridionale were all NIV. The majority of the F. cortaderiae isolates were NIV, with only one 3-ADON genotype. Mycotoxin analysis revealed that none of the samples were contaminated by aflatoxins, OTA, FB2 and type A trichothecenes, however, all of the samples were contaminated with at least one Fusarium toxin. Contamination by DON, ZEA, ENNB and ENNB1 levels were significantly higher in RS. Co-contamination of BEA, DON, ENNs, NIV and ZEA in 18.5% and 24.2% of the analyzed samples was observed, from SP and RS respectively. More than 20% of the samples from RS presented DON and ZEA levels above the regulations established by Europe and Brazil. The results provide further information on the FSAMSC from South America and detected multiple Fusarium toxins in barley samples. This highlights the importance for further studies on the possible interactions of these mycotoxins in order to determine potential risks to animal health.

NIR spectroscopy and chemometric tools to identify high content of deoxynivalenol in barley.

Deoxynivalenol (DON) is one of the mycotoxins produced mainly by the Fusarium graminearum species complex in small grain cereals, including barley. This toxin can cause alimentary disorders, immune function depression and gastroenteritis. The negative health effects associated with DON coupled to the increasing concern about green and rapid methods of analysis motivated this study. In this context, near infrared (NIR) spectroscopy data were applied for exploratory analysis to distinguish barley with high and low levels of DON contamination (> or <1250 µg/kg according to the European Union threshold), by Partial Least Squares-Discriminant Analysis (PLS-DA), and to verify the performance of Partial Least Squares-Regression (PLS-R) to predict DON concentration in barley samples. Maximum values of specificity and sensitivity were achieved in the calibration set; 90.9% and 81.9% were observed in the cross-validation set for the PLS-DA classification model. PLS-R quantification of DON in barley presented low values of error (RMSEC = 101.94 µg/kg and RMSEP = 160.76 µg/kg). Thus, we found that NIR in combination with adequate chemometric tools could be applied as a green technique to monitor DON contamination in barley.

Assessment of Toxigenic Fusarium Species and Their Mycotoxins in Brewing Barley Grains.

Fusarium species threaten yield and quality of cereals worldwide due to their ability to produce mycotoxins and cause plant diseases. Trichothecenes and zearalenone are the most economically significant mycotoxins and are of particular concern in barley, maize and wheat. For this reason, the aim of this study was to characterize the Fusarium isolates from brewing barley and to assess deoxynivalenol and zearalenone contamination in grains. Characterization of the Fusarium strains was carried out by the phylogeny based on two loci (EF-1α and RPB2). Mycotoxin detection and quantification were performed by LC-MS. The results show that Fusarium was the predominant genus. Phylogenetic study demonstrated that the majority of the strains clustered within the Fusarium sambucinum species complex followed by the Fusarium tricinctum species complex. The results revealed high incidence of deoxynivalenol (DON) and zearalenone (ZEA) contamination (90.6% and 87.5%, respectively). It was observed that 86% of the samples contaminated with ZEA were above the limits set by the EU and Brazilian regulations. These results may highlight the importance of controlling Fusarium toxins in barley, mainly because of its use in the brewing industry and the resistance of various mycotoxins to food processing treatments.

Fusarium Mycotoxins Stability during the Malting and Brewing Processes.

Mycotoxins are widely studied by many research groups in all aspects, but the stability of these compounds needs further research for clarification. The objective of this study is to evaluate deoxynivalenol and zearalenone stability during all steps of the malting and brewing processes. The levels of these compounds decreased significantly during the production process (barley to beer). During the malting process, the DON levels decreased significantly in the steeping, germination, and malting steps (62%, 51.5%, and 68%, respectively). Considering ZEN, when the levels were compared between barley and the last step of the process, a significant decrease was observed. Most of the mycotoxins produced were transferred to the rootlets and spent grains, which is advantageous considering the final product. Furthermore, the mycotoxin dietary intake estimation was included in this study. The results proved that if the concentrations of target mycotoxins in raw material are under the limits established by the regulations, the levels decrease during the malting and brewing processes and make the beer secure for consumers. The quality of the five commodities involved in the beer process plays a decisive role in the creation of a safe final product.

Occurrence of deoxynivalenol and zearalenone in brewing barley grains from Brazil.

Barley (Hordeum vulgare L.) is an important cereal crop for food and represents one of the main ingredients in beer production. Considering the importance of barley and its derived products, the knowledge about the mycotoxin contamination in the barley production is essential in order to assess its safety. In this study, the levels of deoxynivalenol (DON) and zearalenone (ZEN) in brewing barley were determined using a LC-MS/MS method. A survey was conducted in 2015 to estimate the mycotoxin levels in these products (n = 76) from four crop regions in Brazil. The results showed high levels of DON and ZEN in the analyzed samples, with contamination levels of 94 and 73.6%, respectively. The mean levels of DON and ZEN ranged from 1700 to 7500 µg/kg and from 300 to 630 µg/kg, respectively. Barley samples from regions 1 and 2 presented higher levels of ZEN and DON, respectively, and those from region 4 presented lower levels of both. Co-occurrence of DON and ZEN was seen in the majority of the barley grain samples, and the mycotoxin content was above the maximum levels established by the Brazilian and European regulations.

Incidence of toxigenic fungi and zearalenone in rice grains from Brazil.

Rice (Oryza sativa L.) is one of the most important food crops worldwide. In Brazil, the southern region is the area with the highest production of rice in the country and also has a high average daily intake of rice by the population. The mycoflora, mainly toxigenic Aspergillus and Fusarium species, the presence of AFB1, DON and ZEA in rice grains, as well as daily intake estimates for the Southern Brazilian population were evaluated. The rice grain samples were collected during the 2017 crop from different harvest periods. According to the mycological tests, the samples presented a high count of fungal colonies in the pre and post-harvest, where the incidence of the F. graminearum species complex (52%) was significantly predominant. This group can be responsible for ZEA production, as found in this study in parboiled rice, mainly because most of the isolated strains were producers of high ZEA levels in the pre-harvest (77%) and post-harvest after natural (79%) and artificial (75%) drying of the rice. Only ZEA showed significant results in the rice grain analyzed (60%) at levels of 90.56 to 126.31 µg/kg, where 36% of the samples were significantly higher than the current maximum limit stipulated in Brazilian regulations and by the European Commission. Despite this, the dietary exposure of ZEA estimated for the southern Brazilian population was below the provisional maximum tolerable daily intake level of 0.5 µg/kg body weight/day set at international regulations.

Mycotoxin analysis of industrial beers from Brazil: The influence of fumonisin B1 and deoxynivalenol in beer quality.

Worldwide, barley is the main source of carbohydrate in the brewing process. However, corn is often used as an adjunct to improve and accelerate the fermentation process. Considering that, these two substrates are susceptible to fungal contamination as well as mycotoxins. The objective of the current study is to determine the incidence of the mycotoxins deoxynivalenol (DON) and fumonisin B1 (FB1) in industrial beers. The method applied for mycotoxin analyses included high performance liquid chromatography. The mean levels for recovery experiments were 89.6% for DON and 93.3% for FB1. DON was not detected in any of the analyzed samples whereas FB1 was found in 49% of the 114 samples. The current survey demonstrated levels of FB1 contamination in industrial beer, possibly due to the addition of contaminated adjuncts. It is necessary to establish maximum levels of mycotoxins in beer in Brazil and other countries in order to reduce health risks.

Degradation of bifenthrin and pirimiphos-methyl residues in stored wheat grains (Triticum aestivum L.) by ozonation.

Pesticide insecticides are used on wheat grains in storage units but their efficiency is hindered by persistent residues in the grains. Therefore, this study aims to evaluate the effectiveness of ozone (O3) gas treatment on the degradation of residual bifenthrin and pirimiphos-methyl insecticides commonly used in storage wheat grains, as well as to evaluate degradation of their by-products. The residues of bifenthrin decreased after 180 min of exposure in a concentration of 60 µmol/mol (a 37.5 ± 7.4% reduction) with 20% moisture content and 0.9 water activity. On the other hand, under the same experimental conditions, the pirimiphos-methyl residues significantly decreased in the wheat grains (71.1 ± 8.6%) after 30 min of exposure. After O3 gas treatment, three by-products of pirimiphos-methyl (m/z=306.1) containing different molecular mass to charge ratios (m/z=278.1, 301.1 and 319.2) were identified by LC-MS. O3 is a strong oxidizer that has shown the potential to reduce pesticide residues in stored grain in order to ensure food quality and safety.

Fumonisins B1 and B2 in the corn-milling process and corn-based products, and evaluation of estimated daily intake.

The distribution of fumonisins (FBs: FB1 and FB2) in the corn-milling process and in corn-based products, as well as daily intake estimates for the Brazilian population were evaluated. Among corn fractions samples, corn meal had the highest mean concentration of FB1 (1305 µg kg(-1)) and FB2 (651 µg kg(-1)) and a distribution factors of 452% and 256% in relation to corn grain, respectively. On the other hand, the distribution factor of FB1 and FB2 in corn flour was found to be 144% and 88% respectively, which demonstrates that fumonisins in this fraction were reduced compared with corn grain. As a result, almost half the corn meal samples (47%) would be non-compliant with future Brazilian regulation (2017) for fumonisins. However, corn-based products, such as corn flakes and popcorn, were in compliance with the regulation. The average probable daily intake and maximum probable daily intake of fumonisins estimated for the Santa Catarina state (Brazil) population were below the provisional maximum tolerable daily intake of 2 µg kg(-1) body weight day(-1) for all corn samples. Despite this, the adoption of practices to control the occurrence of fumonisins should be applied to the corn-milling fractions that may contain a higher concentration of this toxin, such as corn meal, often used for animal feed in Brazil.

Fungi and the natural occurrence of deoxynivalenol and fumonisins in malting barley (Hordeum vulgare L.).

The industrial use of barley grain has experienced continuous growth, mainly due to its economic importance for malt production. From a technological perspective, fungal persistence can reduce product marketability and cause economic losses. In this sense, the aim of the present study was to determine the presence and identification of mycoflora and the occurrence of deoxynivalenol (DON) and fumonisins (FBs) in malting barley. The samples presented a low count of fungal colonies, with values ranging from 10.5 to 0.5 × 10(1)CFU g(-1) and the species most found were Fusarium graminearum and Fusarium verticillioides with 26% and 12% of incidence, respectively. In the samples analyzed for mycotoxins occurrence, DON and FBs were present in 18% and 12%, respectively. The high concentrations of toxins found in the malting samples may be strongly influenced by agricultural practices and the weather conditions during critical phases of plant growth.

Efficacy of zinc compounds in controlling Fusarium head blight and deoxynivalenol formation in wheat (Triticum aestivum L.).

The efficiency of zinc compounds (zinc sulfate, ZnSO4 and zinc oxide, ZnO in regular and nanosize, respectively) on wheat plants was evaluated against growth of Fusarium graminearum and DON formation. In addition, any possible effects on the grain microstructures were observed by scanning electron microscopy (SEM), and the remaining residue of Zn on wheat plants was analyzed. The plants were inoculated with F. graminearum and treated with Zn compounds (100mM) onto spikelets at the anthesis stage. When wheat plants reached maturation, grains were harvested and evaluated for Fusarium (number of colonies, CFU/g), DON formation, and SEM observation, followed by determination of possible remaining Zn residue. The groups treated with ZnSO4 and ZnO-NP showed a reduction in number of CFU of F. graminearum when compared to the control. Similarly for DON formation, i.e. the toxin was reduced to non-detected levels in the treated group. ZnO-NP efficiently reduced F. graminearum and DON formation in the grains at low concentration. Zn remained within the international recommended level for consumption and the treatment did not cause any damage to wheat grains. New strategies of control using Zn compounds in addition to conventional treatments could increase the efficiency against FBH and DON formation.

Mycoflora and deoxynivalenol in whole wheat grains (Triticum aestivum L.) from Southern Brazil.

The fungal species Fusarium graminearum is related to deoxynivalenol (DON) formation. The aim of this study was to evaluate mycoflora and DON occurrence in 53 whole wheat grain samples collected in Southern Brazil during the 2012 crop. Wheat grains showed adequate values of water activity ranging from 0.48 to 0.72, within the required limits of moisture content, ranging from 9.1% to 13.9%. In addition, low counts of fungal colonies, ranging from 10 to 8.2 × 10(2), were found. For Fusarium genera, there was predominance of Fusarium verticillioides (34%) and F graminearum (30.2%). For Aspergillus species, 37.7% of Aspergillus flavus was determined. Regarding the Penicillium species, Penicillium digitatum (49%) was the most found species. DON was detected in 47.2% (25 out of 53) of the samples analysed, with levels ranging from 243.7 to 2281.3 µg kg(-1) (mean: 641.9 µg kg(-1)).