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

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

Risks to human and animal health related to the presence of deoxynivalenol and its acetylated and modified forms in food and feed

Deoxynivalenol (DON) is a mycotoxin primarily produced by Fusarium fungi, occurring predominantly in cereal grains. Following the request of the European Commission, the CONTAM Panel assessed the risk to animal and human health related to DON, 3-acetyl-DON (3-Ac-DON), 15-acetyl-DON (15-Ac-DON) and DON-3-glucoside in food and feed. A total of 27,537, 13,892, 7,270 and 2,266 analytical data for DON, 3-Ac-DON, 15-Ac-DON and DON-3-glucoside, respectively, in food, feed and unprocessed grains collected from 2007 to 2014 were used. For human exposure, grains and grain-based products were main sources, whereas in farm and companion animals, cereal grains, cereal by-products and forage maize contributed most. DON is rapidly absorbed, distributed, and excreted. Since 3-Ac-DON and 15-Ac-DON are largely deacetylated and DON-3-glucoside cleaved in the intestines the same toxic effects as DON can be expected. The TDI of 1 μg/kg bw per day, that was established for DON based on reduced body weight gain in mice, was therefore used as a group-TDI for the sum of DON, 3-Ac-DON, 15-Ac-DON and DON-3-glucoside. In order to assess acute human health risk, epidemiological data from mycotoxicoses were assessed and a group-ARfD of 8 μg/kg bw per eating occasion was calculated. Estimates of acute dietary exposures were below this dose and did not raise a health concern in humans. The estimated mean chronic dietary exposure was above the group-TDI in infants, toddlers and other children, and at high exposure also in adolescents and adults, indicating a potential health concern. Based on estimated mean dietary concentrations in ruminants, poultry, rabbits, dogs and cats, most farmed fish species and horses, adverse effects are not expected. At the high dietary concentrations, there is a potential risk for chronic adverse effects in pigs and fish and for acute adverse effects in cats and farmed mink

Transformation of regulated and emerging mycotoxins upon food processing: from field to digestion

Nel presente lavoro di tesi sono stati studiati diversi aspetti riguardanti micotossine regolate ed emergenti al fine di fornire informazioni circa la loro mitigazione dal campo al prodotto finito, focalizzando l’attenzione sul potenziale impatto del processo tecnologico. Successivamente, il loro ruolo tossicologico, ancora in discussione, è stato indagato attraverso l’applicazione di modelli gastrointestinali. In particolare, lo studio è stato rivolto a DON, micotossina più comune nel grano, DON-3-Glc, che rappresenta la sua principale forma modificata, e alla micotossina ENN B, maggiore rappresentante tra il gruppo delle cosiddette micotossine “emergenti”. Questo studio è stato quindi suddiviso in tre sezioni. La prima è stata rivolta allo sviluppo di esperimenti in serra effettuati su diversi genotipi di grano duro al fine di comprendere meglio il meccanismo di detossificazione dal DON e la resistenza delle diverse varietà ad una delle malattie più gravi che colpiscono questa specie (fusariosi della spiga). La seconda sezione ha riguardato lo studio del reale impatto di alcune filiere di produzione strategiche sul contenuto finale in micotossine. Inoltre, una strategia di mitigazione è stata messa a punto tenendo conto del possibile sviluppo di altri contaminanti legati al processo stesso e ottenendo un prodotto finito adeguato per il consumatore. Nell'ultima sezione di questo lavoro sono invece state effettuate delle indagini circa i possibili risvolti tossicologici di questi composti, valutando il loro destino durante la digestione umana.This work is focused on the study of specific aspects of mycotoxins, regulated and emerging, in order to provide information concerning their mitigation from field to finished product, with a particular focus on the potential impact of food processing. Subsequently, their toxicological role, currently under discussion, has been investigated through a dedicated gastrointestinal model. Particularly, a special focus was devoted to DON, the most common contaminant in wheat, to DON-3-Glc, representing its main derivative, and to ENN B, the major compound among so-called “emerging” mycotoxins. The study has been divided into three sections. The first one is addressed to the development of greenhouse experiments on different durum wheat genotypes in order to better understand the detoxification pathway from DON and the resistance of the different varieties to one of the most severe disease affecting this species. The second section involved the investigation of the effective imoact of different strategic production chains on mycotoxin levels in the finished product. Furthermore, a strategy of mycotoxin mitigation has been approached in order to optimize technological parameters within bakery production, still obtaining an appreciable product for consuming. In the last part of this work, investigations on the toxicological effects of these compounds have been carried out through the evaluation of their fate after human digestion

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

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