Investigating the in vitro catabolic fate of Enniatin B in a human gastrointestinal and colonic model.

Enniatin B is an emerging mycotoxin known to present biological activity because of its ionophoric characteristics. This compound has demonstrated strong in vitro cytotoxicity against different cancer cells, also at low molecular concentrations. Its natural occurrence in food commodities and feed is highly reported world-wide, but few information is available about its stability in the human gastro-intestinal tract. The present work evaluates the catabolic fate of enniatin B upon in vitro simulated digestion and colonic fermentation. LC-MS target and untargeted analysis have been performed to quantify the extent of enniatin B degradation and the formation of catabolic products. The results obtained showed significant degradation of enniatin B (degradation rate 79 ± 5%) along the gastrointestinal tract and further degradation of residual enniatin B was observed during colonic fermentation after 24 h of incubation. Moreover, 5 catabolic metabolites of enniatin B were putatively identified after gastrointestinal digestion resulting from the oxidation and opening of the depsipeptide ring. As a final step, the pharmacokinetic properties of enniatin B degradation products were tested in silico revealing that some of them may be adsorbed at the gastrointestinal level more than the parent compound. Additionally, the smaller degradation products showed moderate blood-brain-barrier crossing.

Acrylamide Reduction Strategy in Combination with Deoxynivalenol Mitigation in Industrial Biscuits Production.

Acrylamide is formed during baking in some frequently consumed food products. It is proven to be carcinogenic in rodents and a probable human carcinogen. Thus, the food industry is working to find solutions to minimize its formation during processing. To better understand the sources of its formation, the present study is aimed at investigating how acrylamide concentration may be influenced by bakery-making parameters within a parallel strategy of mycotoxin mitigation (focusing specifically on deoxynivalenol-DON) related to wholegrain and cocoa biscuit production. Among Fusarium toxins, DON is considered the most important contaminant in wheat and related bakery products, such as biscuits, due to its widespread occurrence. Exploiting the power of a Design of Experiments (DoE), several conditions were varied as mycotoxin contamination levels of the raw materials, recipe formulation, pH value of dough, and baking time/temperature; each selected treatment was varied within a defined range according to the technological requirements to obtain an appreciable product for consumers. Experiments were performed in a pilot-plant scale in order to simulate an industrial production and samples were extracted and analysed by HPLC-MS/MS system. Applying a baking temperature of 200 °C at the highest sugar dose, acrylamide increased its concentration, and in particular, levels ranged from 306 ± 16 µg/Kg d.m. and 400 ± 27 µg/Kg d.m. in biscuits made without and with the addition of cocoa, respectively. Conversely, using a baking temperature of 180 °C in the same conditions (pH, baking time, and sugar concentrations), acrylamide values remained below 125 ± 14 µg/Kg d.m. and 156 ± 15 µg/Kg d.m. in the two final products. The developed predictive model suggested how some parameters can concretely contribute to limit acrylamide formation in the final product, highlighting a significant role of pH value (correlated also to sodium bicarbonate raising agent), followed by baking time/temperature parameters. In particular, the increasing range of baking conditions influenced in a limited way the final acrylamide content within the parallel effective range of DON reduction. The study represents a concrete example of how the control and optimization of selected operative parameters may lead to multiple mitigation of specific natural/process contaminants in the final food products, though still remaining in the sensorial satisfactory range.

Formulation and processing factors affecting trichothecene mycotoxins within industrial biscuit-making.

Food processing, especially thermal treatment, may have implications on mycotoxins in products available for consumers. This research work aimed to study how mycotoxin levels may be influenced by modifying the technological parameters of both whole grain and cocoa biscuit-making processes. The study was mainly focused on the following mycotoxins: deoxynivalenol, deoxynivalenol-3-glucoside, and the minor metabolite culmorin. Special emphasis was given to the recipe formulation, and to the baking conditions, using an industrial-scale operation, starting from naturally contaminated raw materials. Exploiting the power of Design of Experiments (DoE) and a dedicated LC-MS/MS method, the complexity of the different processes was investigated. The models obtained within this study showed a high goodness-of-fit suggesting that the pH and the baking time play important roles for minimizing mycotoxins in the final products, while the recipe formulation has an impact on the mycotoxins extractability by affecting the biscuit microstructure.

Deoxynivalenol & Deoxynivalenol-3-Glucoside Mitigation through Bakery Production Strategies: Effective Experimental Design within Industrial Rusk-Making Technology.

In the scientific field, there is a progressive awareness about the potential implications of food processing on mycotoxins especially concerning thermal treatments. High temperatures may cause, in fact, transformation or degradation of these compounds. This work is aimed to study the fate of mycotoxins during bakery processing, focusing on deoxynivalenol (DON) and deoxynivalenol-3-glucoside (DON3Glc), along the chain of industrial rusk production. Starting from naturally contaminated bran, we studied how concentrations of DON and DON3Glc are influenced by modifying ingredients and operative conditions. The experiments were performed using statistical Design of Experiment (DoE) schemes to synergistically explore the relationship between mycotoxin reduction and the indicated processing transformation parameters. All samples collected during pilot plant experiments were analyzed with an LC-MS/MS multimycotoxin method. The obtained model shows a good fitting, giving back relevant information in terms of optimization of the industrial production process, in particular suggesting that time and temperature in baking and toasting steps are highly relevant for minimizing mycotoxin level in rusks. A reduction up to 30% for DON and DON3Glc content in the finished product was observed within an acceptable technological range.

Effects of orally administered fumonisin B1 (FB1), partially hydrolysed FB1, hydrolysed FB1 and N-(1-deoxy-D-fructos-1-yl) FB1 on the sphingolipid metabolism in rats.

Fumonisin B1 (FB1) is a Fusarium mycotoxin frequently occurring in maize-based food and feed. Alkaline processing like nixtamalisation of maize generates partially and fully hydrolysed FB1 (pHFB1 and HFB1) and thermal treatment in the presence of reducing sugars leads to formation of N-(1-deoxy-D-fructos-1-yl) fumonisin B1 (NDF). The toxicity of these metabolites, in particular their effect on the sphingolipid metabolism, is either unknown or discussed controversially. We produced high purity FB1, pHFB1a+b, HFB1 and NDF and fed them to male Sprague Dawley rats for three weeks. Once a week, urine and faeces samples were collected over 24 h and analysed for fumonisin metabolites as well as for the sphinganine (Sa) to sphingosine (So) ratio by validated LC-MS/MS based methods. While the latter was significantly increased in the FB1 positive control group, the Sa/So ratios of the partially and fully hydrolysed fumonisins were indifferent from the negative control group. Although NDF was partly cleaved during digestion, the liberated amounts of FB1 did not raise the Sa/So ratio. These results show that the investigated alkaline and thermal processing products of FB1 were, at the tested concentrations, non-toxic for rats, and suggest that according food processing can reduce fumonisin toxicity for humans.

Fusarium species, chemotype characterisation and trichothecene contamination of durum and soft wheat in an area of central Italy.

BACKGROUND: Fusarium head blight (FHB) of wheat is an important disease causing yield losses and mycotoxin contamination. The aim of the work was to detect and characterise trichothecene producing Fusarium species in durum and soft wheat cultivated in an area of central Italy in 2009 and 2010 and to determine trichothecene contamination by LC-MS/MS in the grain. RESULTS: F. graminearum s. str. was the most frequent species. In 2009, the occurrence of F. avenaceum and F. poae was higher than in 2010. Among F. graminearum strains, the 15-acetyl deoxynivalenol (15-ADON) chemotype could be found more frequently, followed by nivalenol (NIV) and 3-ADON chemotypes, while all F. culmorum isolates belonged to the 3-ADON chemotype. All F. poae strains were NIV chemotypes. In vitro trichothecene production confirmed molecular characterisation. Durum wheat was characterised by a higher average DON contamination with respect to soft wheat, NIV was always detected at appreciable levels while type-A trichothecenes were mostly found in durum wheat samples in 2009 with 6% of samples exceeding the contamination level recently recommended by the European Union. CONCLUSION: Climatic conditions were confirmed to be predominant factors influencing mycotoxigenic species composition and mycotoxin contaminations. However, NIV contamination was found to occur irrespective of climatic conditions, suggesting that it may often represent an under-estimated risk to be further investigated.

Durum wheat (Triticum Durum Desf.) lines show different abilities to form masked mycotoxins under greenhouse conditions.

Deoxynivalenol (DON) is the most prevalent trichothecene in Europe and its occurrence is associated with infections of Fusarium graminearum and F. culmorum, causal agents of Fusarium head blight (FHB) on wheat. Resistance to FHB is a complex character and high variability occurs in the relationship between DON content and FHB incidence. DON conjugation to glucose (DON-3-glucoside, D3G) is the primary plant mechanism for resistance towards DON accumulation. Although this mechanism has been already described in bread wheat and barley, no data are reported so far about durum wheat, a key cereal in the pasta production chain. To address this issue, the ability of durum wheat to detoxify and convert deoxynivalenol into D3G was studied under greenhouse controlled conditions. Four durum wheat varieties (Svevo, Claudio, Kofa and Neodur) were assessed for DON-D3G conversion; Sumai 3, a bread wheat variety carrying a major QTL for FHB resistance (QFhs.ndsu-3B), was used as a positive control. Data reported hereby clearly demonstrate the ability of durum wheat to convert deoxynivalenol into its conjugated form, D3G.