Suchergebnisse

Owing to their conditions of production, and sometimes to poor storage and transportation conditions, herbal products are susceptible to fungal contamination and development. This can lead to the accumulation of mycotoxins in this kind of commodities. Consequently, herbal products can be contaminated with fungal toxins, such as aflatoxins, ochratoxins and other mycotoxins, which pose a serious risk to public health. This paper reviews the main aspects regarding mycotoxin contamination of medical/aromatic herbs, in the context of the importance of this product in a global market. Moreover, the effect of processing on final contamination of derived foods, as well as the analytical methodology commonly employed in fungal and mycotoxin analysis in this type of products was reviewed. ; Debido a sus condiciones habituales de producción, así como a condiciones inadecuadas de almacenamiento y de transporte, los productos a base de plantas medicinales o aromáticas son susceptibles a la contaminación por mohos, lo que puede conducir a la acumulación de toxinas fúngicas tales como las aflatoxinas, ocratoxinas y otras micotoxinas, lo que supone un grave riesgo para la salud pública. Este artículo revisa los principales aspectos de la contaminación por micotoxinas de las hierbas medicinales y/o aromáticas, teniendo en cuenta el contexto de este producto en un mercado globalizado, así como el efecto del procesado sobre la contaminación final de estos productos. También se revisa la metodología más comúnmente empleada en el análisis de mohos y micotoxinas en este tipo de productos. ; The authors are grateful to the Spanish (Project AGL2007-66416-C05) and Catalonian (XaRTA- Reference Network on Food Technology) Government for their financial support.

In 2009 the EU Regulation 386/2009 established a new functional group of feed additives called 'substances for reduction of the contamination of feed by mycotoxin'. Later, di-octahedral bentonite (1 m558) was authorised, as an anti-aflatoxin additive, being the only additive of this group authorised to date. This work aims to demonstrate the effectiveness of other bentonites, such as tri-octahedral bentonites, versus Fusarium-mycotoxins, since very few adsorbents have proved their effectiveness in relation to this group of mycotoxins. For this purpose, 7 bentonites (six of them tri-octahedral) and 7 commercial adsorbents, added at 0.02% (w/v), were assayed in an in vitro adsorption experiment using two simulated gastro-intestinal (GI) juices (pH 1.3 and 6.8) versus zearalenone (ZEN: 0.1-5 mg/L), fumonisin B1 (FB1: 1-10 mg/L) and deoxynivalenol (DON: 2-10 mg/L). Mycotoxin adsorption data were fitted to Langmuir and Freundlich isotherms. In vitro adsorption experiments showed that ZEN and FB1 (in the latter case only in acid medium) were partially adsorbed, while the adsorption of DON was negligible. Moreover, the increase of adsorbent dose (up to 0.20%, w/v) significantly improved the in vitro adsorption of ZEN and FB1, reaching >90% of adsorption. The present work proposes the use of some tri-octahedral bentonites as feed additives for Fusarium-mycotoxin reduction. ; The authors are grateful to the following institutions: Spanish government (projects AGL2014-52648-REDT and RTC-2015-3508-2–ERDF. A way to make Europe, funded by the Ministry of Economy and Competitiveness of the Government of Spain and by the European Regional Development Fund of the European Union with the objective of "Promoting technological development, innovation and quality research") and to XaRTA (The Catalonian Reference Network in Food Technology) for funding. P. Vila-Donat thanks MINECO for her postdoctoral contract.

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

The stability of deoxynivalenol (DON), deoxynivalenol-3-glucoside (DON-3-glucoside), 3-acetyldeoxynivalenol (3-ADON), 15-acetyldeoxynivalenol (15-ADON), de-epoxy-deoxynivalenol (DOM-1) and ochratoxin A (OTA) during thermal processing has been studied. Baking temperature, time and initial mycotoxin concentration in the raw materials were assayed as factors. An improved UPLC-MS/MS method to detect DON, DON-3-glucoside, 3-ADON, 15-ADON and DOM-1 in wheat baked products was developed in the present assay. The results highlighted the importance of temperature and time in mycotoxin stability in heat treatments. OTA is more stable than DON in a baking treatment. Interestingly, the DON-3-glucoside concentrations increased (>300%) under mild baking conditions. On the other hand, it was rapidly reduced under harsh conditions. The 3-ADON decreased during the heat treatment; while DOM-1 increased after the heating process. Finally, the data followed first order kinetics for analysed mycotoxins and thermal constant rates (k) were calculated. This parameter can be a useful tool for prediction of mycotoxin levels. ; The authors are grateful to the Spanish Government (projects AGL2010-22182-C04-04 and AGL2011-24862) for the financial support. A. Vidal thanks the Spanish Government (Ministry of Education) for the pre-doctoral grant.

The mycotoxin deoxynivalenol (DON) is one of the most common mycotoxins of cereals worldwide, and its occurrence has been widely reported in raw wheat. The free mycotoxin form is not the only route of exposure; modified forms can also be present in cereal products. Deoxynivalenol-3-glucoside (DON-3-glucoside) is a common DON plant conjugate. The mycotoxin concentration could be affected by food processing; here, we studied the stability of DON and DON-3-glucoside during baking of small doughs made from white wheat flour and other ingredients. A range of common food additives and ingredients were added to assess possible interference: ascorbic acid (E300), citric acid (E330), sorbic acid (E200), calcium propionate (E282), lecithin (E322), diacetyltartaric acid esters of fatty acid mono- and diglycerides (E472a), calcium phosphate (E341), disodium diphosphate (E450i), xanthan gum (E415), polydextrose (E1200), sorbitol (E420i), sodium bicarbonate (E500i), wheat gluten and malt flour. The DON content was reduced by 40%, and the DON-3-glucoside concentration increased by >100%, after baking for 20 min at 180°C. This confirmed that DON and DON-3-glucoside concentrations can vary during heating, and DON-3-glucoside could even increase after baking. However, DON and DON- 3-glucoside are not affected significantly by the presence of the food additives tested. ; The authors are grateful to the Spanish government (project AGL2014-55379-P) for providing financial support. A. Vidal thanks the Spanish government (Ministry of Education) for the pre-doctoral grant.

The objective of this study was to characterize the mycoflora of sorghum grains commercialized in the Tunisian retail market and to identify aflatoxins (AFs), ochratoxin A (OTA) and zearalenone (ZEA) producing species. Sixty four samples of sorghum (37 samples of Tunisian sorghum and 27 samples of Egyptian sorghum) were analyzed. Dilution plating (CFU, colony forming units) was used for fungal enumeration. The isolation of mycobiota was carried out by plating of grains on PDA and malachite green medium. Aspergillus section Flavi and section Nigri and Fusarium isolates were sub-cultured in CYA to test their ability to produce AFs, OTA and ZEA, respectively. The selected Aspergillus section Flavi and section Nigri, Penicillium and Fusarium isolates were subjected to specific PCR assays using published species-specific primers. The results revealed the dominance of Fusarium (95.3%), followed by Aspergillus (87.2%) and Alternaria (81.2%) species. The fungal counts ranged from 100 to 1.3·104 CFU/g for Tunisian sorghum and from 100 to 5.7·103 CFU/g for Egyptian sorghum. Among Aspergillus section Flavi isolates identified by molecular biology, Aspergillus flavus was the most dominant (90.1%) while Aspergillus parasiticus represent 9.9% only. About Aspergillus section Nigri, results showed the dominance of Aspergillus niger aggregate species, including Aspergillus niger, Aspergillus tubingensis and other species. Among Fusarium species, Fusarium incarnatum was the most dominant in both Tunisian and Egyptian sorghum. Penicillium citrinum was the dominant Penicillium species in the studied samples. More than 890 isolates belonging to the genus Aspergillus and Fusarium were tested in order to test their capacity to produce AFs, OTA and ZEA. The percentage of mycotoxin producing isolates in Aspergillus section Flavi, A. section Nigri, and Fusarium was 30.0%, 4.6% and 11.1%, respectively. ; We thank Montse Prim for her encouragement and technical assistance. The authors are grateful to the European Union (MYCORED KBBE-2007-2-5-05 project) and Tunisian government for the financial support.

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

A study was undertaken to evaluate the impact of the application of several fungicide treatments used in Spanish vines on Aspergillus carbonarius growth and ochratoxin A production. Three trials were designed in order: (1) to screen 26 fungicides at the doses recommended by manufacturers on grape-like synthetic medium at 20 and 30 C; (2) to find out the minimum inhibitory concentration of each fungicide for A. carbonarius growth on synthetic medium; and (3) to investigate the effect of several fungicides on A. carbonariu s-inoculated grapes. In synthetic medium nine fungicides significantly reduced A. carbonarius growth rate. Meanwhile, 13 fungicides completely inhibited its growth. In general, growth was faster at 30 C than at 20 C, contrary to ochratoxin A production. Fungicides that stopped fungal growth also inhibited ochratoxin A production, but not all the fungicides that reduced growth reduced the ochratoxin A synthesis. In general, fungicides that contained copper or strobilurins reduced both growth and ochratoxin A production, contrary to sulphur fungicides. At the optimum temperature for A. carbonarius growth of 30 C, higher amounts of fungicide were needed to prevent fungal growth than at 20 C. Among the fungicides that inhibited A. carbonarius growth on synthetic medium at the initial doses, cyprodinil seemed to be the active ingredient more effective at stopping fungal growth when testing reduced doses. The fungicide effect on grapes was similar to that on synthetic medium. Both infection and ochratoxin A production were reduced when using cyprodinil (37.5%) plus fludioxonil (25%) and azoxystrobin (25%). Penconazole (10%) also showed a clear reduction in ochratoxin A production at both temperatures, although infection was only reduced at 20 C. Ochratoxin A reduction was strain and temperature-dependent. In general, fenhexamid (50%), mancozeb (80%) and copper hydroxide (80%) plus copper (50%) enhanced infection and ochratoxin A production. ; The work was funded by the Catalonian Government (Direcció General de Recerca, Generalitat de Catalunya), the Spanish Government (CICYT, Comisión Interministerial de Ciencia y Tecnologí, AGL2004-07549-C05-01, and Ramon y Cajal programme) and the European Commission, Quality of Life Programme (QoL), Key Action 1 (KA1) on Food, Nutrition and Health (QLRT- 2000-01761). The authors are grateful to Syngenta- Agro S.A. (Spain) and Le ́ rida Unio ́ n Quı ́ mica S.A. (Luqsa, Spain) for providing the fungicides used

Deoxynivalenol (DON) is one of the most frequently occurring mycotoxins in wheat crops worldwide and poses a risk to human and animal health due to its wide range of adverse effects. Deoxynivalenol-3-glucoside (DON-3-glucoside) is a DON plant conjugate that is widely found in cereal products. As DON accumulation in the field seems unavoidable, it is important to investigate all of the conditions that affect its stability during food processing. One of the most consumed cereal product around the world is bread, however the published information about DON stability in bread shows a large variability of results because a huge amount of factors affect DON and its modified forms. So, the aim of this research was to study the fate of DON and its modified forms through the breadmaking process with the addition of xylanase and α-amylase at different fermentation temperatures. Moreover, different α-amylase and xylanase concentrations were added to the dough to be fermented. To quantify DON and its derived forms in the samples, liquid chromatography with double mass spectrophotometer was used. DON was reduced during fermentation and baking; however, the reduction at each step was related to the fermentation temperature. The presence of α-amylase and xylanase caused increases in DON during fermentation and during early baking. DON-3-glucoside was slightly reduced after fermentation and was widely increased (>80%) after baking. Deepoxy-deoxynivalenol (DOM-1) increased during the breadmaking process. Breadmaking process can reduce DON concentration, however xylanase and α-amylase presence cause increases of DON. ; The authors are grateful to the Spanish government (project AGL2014-55379-P) for financial support. A. Vidal thanks the Spanish government (Ministry of Education) for the pre-doctoral grant.

The main mycotoxigenic fungi found in maize in the Mediterranean basin include Aspergillus section Flavi and several Fusarium species, Fusarium section Liseola being the more widespread. While Aspergillus section Flavi species can develop in the field or in stored maize in Mediterranean countries, Fusarium species colonise maize ears in the field. As a consequence, fumonisins are the major contaminants in Mediterranean maize, together with aflatoxins. The prevalence of Fusarium section Liseola in the Mediterranean countries is closely linked to the activity of insects such as Sesamia nonagrioides and Ostrinia nubilalis. The incidence of the different mycotoxigenic fungi and mycotoxins across the Mediterranean countries is extensively reviewed in this work. Furthermore, both pre- and postharvest strategies to reduce the presence of such toxins in the maize chain are described. Finally, the incidence and levels of mycotoxins encountered in maize products intended for direct human consumption in the Mediterranean countries are also assessed; they are much lower than those in maize grain as a result of food processing technologies. ; The authors wish to thank to FP7 EU project MycoRed (GA222690) for giving the opportunity to prepare this review article as well as to the Spanish Government (project AGL2010-22182-C04-04) for the financial support.

Novel strategies that address vitamin A deficiency have been developed, such as high-carotenoid maize, a biofortified transgenic maize line rich in carotenoids generated by genetic transformation. The South African white maize inbred (M37W), which is devoid of carotenoids, was engineered to accumulate high levels of β-carotene (provitamin A), lutein, and zeaxanthin. Maize seeds contaminated with fumonisins and other mycotoxins pose a serious threat to both humans and livestock. During three consecutive harvests, the fungal incidence and the fumonisin and aflatoxin content of maize seeds grown in an experimental field in Catalonia (Northeastern Spain) were evaluated. Fungal infection was similar in high-carotenoid maize and its isogenic line, with Fusarium verticillioides being the most prevalent fungus in all the harvests. Neither Aspergillus spp. nor aflatoxin contamination was found. Fumonisin levels were lower in high carotenoid than in its isogenic line, but this reduction was statistically significant in only 2 of the 3 years of study. Our results suggest that high carotenoid content reduces fumonisin levels in maize grains. ; The study was supported by La Caixa (Recercaixa project PC084082 VitaMaize: high-quality and safe food through antioxidant fortified maize), the Agrotecnio Center, the Catalonian Government (2014 SGR 1296 Agricultural Biotechnology Research Group and XaRTA Reference Network on Food Technology) and the Spanish Ministry of Economy and Competitiveness (BIO2014-54441-P). J. Díaz-Gómez thanks the University of Lleida for a pre-doctoral grant. The authors would like to thank the Agricultural Biotechnology group of the University of Lleida, Antonio Michelena, and Jaume Lloveras for their work in the experimental field.

Aflatoxin M1 (AFM1) is a hydroxylated metabolite of aflatoxin B1 (AFB1) and can be excreted in milk of cows after consuming aflatoxin (AF)-contaminated feed. The aim of this research was to assess the levels of total AFs in samples of feedingstuff for dairy cows (n = 193) and the levels of AFM1 in raw bulk tank milk samples (n = 375), in order to estimate the ratio of BAFB1 feed input^ versus BAFM1 milk output^ in four specific regions of Spain. Moreover, the correlation between the raw materials used as ingredients of the total mixed ration (TMR) and the presence of AFs was studied. About one-third (34.7%) of the feed samples were positive for total AFs in a range of 0.05-6.45 μg/kg, and 12.4% were positive for AFB1. AFM1 was detected in 18.9% of bulk milk samples, with concentrations ranging from 0.009 to 1.36 μg/kg. While none of the feed samples exceeded the European Union (EU) maximumcontent for AFB1 in feedingstuff for dairy animals of 5 μg/kg, three bulk milk samples exceeded the EU maximum level for AFM1 in milk of 50 ng/kg. The transfer ratio AFB1/AFM1, which was derived from AFB1 levels in feed, AFM1 levels in bulk tank milk, feed intake, and milk yield data, was 0.6-6%, which corresponded well with the range of published carry-over data for aflatoxins. Statistical analyses showed that the main sources of AFB1 in TMR were maize silage, bagasse, soya bean husk, maize, alfalfa hay, cotton seed and compound feed, thus special attention should be paid in controlling these raw materials when used in TMR preparation. Although the analysis of AFs in feed did not correlate with the presence of AFM1 in milk, monitoring feedstuffs is a useful tool in order to try and minimise AF-contamination of milk. ; This work was supported by the Spanish Ministry of Economy and Competitiveness (project AGL2014-55379-P). M. Rodríguez-Blanco thanks the University of Lleida for a pre-doctoral grant.

The fate of deoxynivalenol (DON) and ochratoxin A (OTA) during the breadmaking process was studied. In particular, toxin content was analysed in mixed baking ingredients before kneading, after fermentation and proofing, and finally after baking. Fermentation and proofing were carried out at 30 °C for 1 h, while baking was performed at different temperature levels (from 170 to 210 °C) and baking times from 45 to 135 min, in a full factorial design. DON increased from unkneaded mix to fermented dough, and decreased due to baking; this trend depended on the initial concentration of DON in the flour. The level in the bread was significantly lower than in the initial mix of ingredients. In contrast, deoxynivalenol-3-glucoside (DON-3-G) content increased both during kneading and fermentation, and also during baking. Moreover, the results confirmed the high stability of OTA as no significant change in its content could be observed as a result of the breadmaking process. As conclusion, the design of bakery product processes may help to control DON in final products, because although quite stable, its levels can be reduced to some extent. However, high levels of DON-3-G were released during baking, and this point should be further investigated. Mycotoxins have been always considered as stable compounds; however, in depth knowledge of the processing steps that may lead to some reduction (although limited) and those which can stimulate their release from conjugated forms, will definitely help in their control in finished foodstuffs. ; The authors are grateful to the Spanish government (projects AGL2010-22182-C04-04 and AGL2011-24862) for the financial support. A. Vidal thanks the Spanish Government (Ministry of Education) for the pre-doctoral grant. H. Morales is grateful to the Portuguese Government. (Ministério da Ciêcia, Tecnologia e Ensino Superior; FCT Fundação para a Ciência e a Tecnologia) Grant ref. SFRH/BPD/38011/2007.

Deoxynivalenol (DON) and ochratoxin A (OTA) are mycotoxins produced by fungal species which can contaminate, alone or simultaneously, cereal-based products such as bread. Due to the increasing interest in the beneficial effects of dietary bran, bran bread has attained high consumption. Usually, the higher mycotoxin concentrations in cereals are found in the external layers of the grain (bran), leading to higher concentration of DON and OTA in breads with added bran. Moreover, the use of sourdough in breadmaking is increasing, but no studies about its effect in the mycotoxins content exist. The objective of this study was to determine the variation of concentration of these mycotoxins during the breadmaking process including the following factors: two initial mycotoxin concentrations in the initial mix of ingredients, four different bran contents, and use of sourdough. OTA was confirmed to be quite stable during the breadmaking process, regardless of the assayed factors. DON concentration during breadmaking was not significantly affected by bran content of bread. However, it was significantly affected by kneading and fermentation steps in opposite way depending on sourdough use and flour contamination level: if DON reduction occurs during fermentation, this leads to a safer situation, but the possible increase in DON should be considered with care, as it can compensate the expected dilution effect by recipe. Finally, the results on deoxynivalenol-3-glucoside (DON-3-G), although preliminar, suggest an increase of this toxin during fermentation, but mainly during baking. ; The authors are grateful to the Spanish Government (Projects AGL2010-22182-C04-04 and AGL2011-24862) for the financial support. A. Vidal thanks the Spanish Government (Ministry of Education) for the pre-doctoral grant. H. Morales is grateful to the Portuguese Government (Ministério da Ciência, Tecnologia e Ensino Superior; FCT Fundação para a Ciência e a Tecnologia) Grant Ref. SFRH/BPD/38011/2007.

The occurrence of two Alternaria mycotoxins, alternariol (AOH) and alternariol monomethyl ether (AME) and the presence of conidia from Alternaria spp., were investigated throughout the food production chain of two businesses, one which uses organic fruit and the other non-organic. For this purpose, a propidium monoazide (PMA) treatment followed by a quantitative Real Time PCR (qPCR) was used to detect and quantify viable conidia exclusively. Results demonstrated that 68.4% of the total raw fruit analysed was contaminated with viable Alternaria spp. Regarding the mycotoxin occurrence, only a few samples were contaminated with AME, while 35% of raw tomatoes tested positive for AOH in the organic producer and 21% in the non-organic producer. AOH was present in samples analysed before heat treatment, while almost no mycotoxins were found in the final products of the organic producer. However, in the non-organic producer, 47% of the tomato concentrates were contaminated. ; We thank both companies for their collaboration in this study and for opening their doors to us. We also thank Montse Prim for kindly helping us with the HPLC analysis of samples and Arthur Kendall for correction of English usage and style. We are grateful to the Catalonian Government (XaRTA-Reference Network on Food Technology) and the University of Lleida for their financial support. N. Estiarte thanks the Secretaria de Universitats i Recerca del Departament de Economia i Coneixement of the Generalitat de Catalunya for the pre-doctoral grant (FI-DGR 2013).