Les mycotoxines sont des métabolites secondaires des moisissures qui peuvent naturellement contaminer de nombreux supports (céréales, fruits, papiers peints, compost). Dans ces travaux de thèse, nous avons focalisé notre intérêt sur deux mycotoxines produites principalement par les champignons du genre Fusarium, appartenant toutes deux au même groupe des trichothécènes, le déoxynivalénol (DON) et la toxine T-2 (T-2). Les objectifs de cette thèse ont été de déterminer les effets de ces deux toxines sur la mise en place des réponses immunitaires au sein de la muqueuse respiratoire et intestinale. L’ensemble de ces études a été réalisé chez le porc, espèce cible de ces contaminants et animal modèle pour l’Homme. Les résultats de ces travaux ont montré que ces deux toxines affectent la réponse immunitaire. Au niveau du tractus respiratoire, une faible dose de toxine T-2 altère l’activation des macrophages alvéolaires lorsqu’ils sont stimulés par les agonistes des TLRs -4, -2/6 (lipopolysaccharides et acides lipoteichoïques, respectivement) en diminuant la synthèse du composé antimicrobien NO et des cytokines pro-inflammatoires IL-1β et TNF-α. Cette immunosuppression pourrait alors conduire à une susceptibilité plus accrue des porcs à des infections opportunistes. Au niveau du tractus intestinal, à l’état basal, nous avons mis en évidence que le DON comme la toxine T-2 induit une forte réponse inflammatoire innée associée à la stimulation de la voie IL-17 en inhibant le développement des cellules T régulatrices. Des études mécanistiques ont permis de déterminer que les cytokines associées à la voie IL-17 suite à une exposition aux trichothécènes, sont produites par une des sous-populations de cellules T innées, les cellules Tγδ productrices d’IL-17. La troisième partie de ce travail a porté sur l’utilisation de la souche Lactobacillus sobrius DSM 16698T dans le but de lutter contre les effets immunomodulateurs générés lors d’une exposition aux trichothécènes. Les résultats de ce travail ont montré que cette souche bactérienne est capable de réduire les effets inflammatoires IL-17 et de rétablir les paramètres impliqués dans les fonctions de la barrière intestinale, suite à une exposition ex vivo et in vivo au DON. En revanche, cette souche a peu d’effet contre la toxine T-2. L’ensemble de ce travail de thèse suggère donc qu’une exposition à de faibles doses de trichothécènes pourrait accroître la susceptibilité des animaux aux perturbations de nature infectieuse ou inflammatoire. / Mycotoxins are fungi secondary metabolites that can contaminate a lot of environments worldwild such as cereals, fruits, wallpapers, and compost heaps. Throughout this phD work, we focused on two mycotoxins mainly produced by Fusarium species, both belonging to the trichothecenes group: the deoxynivalenol (DON) and the T-2 toxin (T-2). The aims of this study were to determine the effects of these two toxins on the immune response implementation in respiratory and intestinal mucosa. Studies were performed on swine being a target species of these contaminants and a model species for Humans. The results of these works proved that these two mycotoxins can affect the immune response. In the respiratory tract, a low dose of T-2 toxin alters the activation of the alveolar macrophages when they are stimulated by the agonists of TLRs -4 and -2/6 (lipopolysaccharides and lipoteichoic acids, respectively). This alteration is due to the decrease of the synthesis of the anti-microbial compound NO and the pro-inflammatory cytokines such as IL-1β and TNF-α. This immunosuppression can induce the emergence of opportunist infections in pig. In the intestinal tract, in background level, we demonstrated that DON as well as T-2 toxin induces a strong inflammatory immune response associated with stimulation of IL-17 pathway by inhibiting of the development of regulatory T cells. Mechanistic studies were used to determine the production origin of the cytokin associated to the IL-17 pathway. This cytokine is produced by one of the subpopulations of Tregs, the Tγδ cells IL-17 producing when exposed to trichothecenes. The third part of this work was about the use of Lactobacillus sobrius DSM 16698T strain to counteract the immunomodulatory effects induced after trichothecen exposure. The results of this study showed that this bacterial strain is able to reduce IL-17 inflammatory effect and is also able to re-etablish the parameters involved in the intestinal barrier functions in ex vivo and in vivo response to DON. Nevertheless, this strain is less effective against the T-2 toxin. Taken together, results of this phD suggest that an exposure to low doses of trichothecens could be intensify the susceptibility of animal to infectious or inflammatory disease.

Maize is the principal staple food/feed crop exposed to mycotoxins, and the co-occurrence of multiple mycotoxins and their metabolites has been well documented. Dietary (co)-exposure to mycotoxins is associated with human and animal health concerns as well as economic losses. The present thesis aims to apply a holistic approach for the risk assessment of mycotoxin mixtures in food and feed, i.e. from fungal production and occurrence to harmonised risk characterisation. This was done in three folds. Firstly, available environmental, ecological, and agronomic factors that may affect the relative abundance of co-occurring mycotoxins in the contaminated crops were collected from peer-reviewed literature, with focus on maize (Chapter I). Secondly, (co-)occurrence data on mycotoxins in core cereals was extracted from available articles in the scientific literature and analysed to estimate potential pattern of co-exposure in humans and animals (Chapter II). Finally, Chapter III investigates the applicability of the EFSA guidance to multiple mycotoxins through a scenario of possible co-exposure in humans and animals, using maize as a case study. In particular, a human and animal risk assessment to mycotoxin mixture in maize was conducted using a modelled component-based approach for selected mixture of mycotoxins, that, according to our data, co-occur in maize based feed and food products. / Maize is the principal staple food/feed crop exposed to mycotoxins, and the co-occurrence of multiple mycotoxins and their metabolites has been well documented. Dietary (co)-exposure to mycotoxins is associated with human and animal health concerns as well as economic losses. The present thesis aims to apply a holistic approach for the risk assessment of mycotoxin mixtures in food and feed, i.e. from fungal production and occurrence to harmonised risk characterisation. This was done in three folds. Firstly, available environmental, ecological, and agronomic factors that may affect the relative abundance of co-occurring mycotoxins in the contaminated crops were collected from peer-reviewed literature, with focus on maize (Chapter I). Secondly, (co-)occurrence data on mycotoxins in core cereals was extracted from available articles in the scientific literature and analysed to estimate potential pattern of co-exposure in humans and animals (Chapter II). Finally, Chapter III investigates the applicability of the EFSA guidance to multiple mycotoxins through a scenario of possible co-exposure in humans and animals, using maize as a case study. In particular, a human and animal risk assessment to mycotoxin mixture in maize was conducted using a modelled component-based approach for selected mixture of mycotoxins, that, according to our data, co-occur in maize based feed and food products.

Le déoxynivalénol (DON) est l'une des mycotoxines trichothécènes les plus abondantes et les plus importantes produites par les espèces de Fusarium. Les aliments contaminés par les mycotoxines peuvent affecter négativement la santé des poulets de chair. La présente étude a été menée pour évaluer les effets du déoxynivalénol (DON) sur les performances des poulets de chair, le poids des organes, les paramètres biochimiques sériques, la réponse immunitaire et l'histologie intestinale. L’expérience a permis d’évaluer l'efficacité d'un additif alimentaire commercial, basé sur les synbiotiques (SFA), et ayant la capacité d'oxyder le DON. Au total, six cents poussins mâles d'un jour d'une souche commerciale (Ross 308) ont été vaccinés avec un vaccin vivant combiné contre le virus de la maladie de Newcastle (NDV) et le virus de la bronchite infectieuse (IBV) et ont été répartis de façon aléatoire entre 6 traitements (100 oiseaux dans chaque groupe) pour 5 semaines. Ces groupes ont été nourris avec la même nourritures naturellement contaminés par DON mais à des concentrations différentes : < 0,5 (régime témoin), 1,5 et 3 mg/kg avec et sans SFA à 250 g/tonne. Les paramètres, y compris le poids corporel, la consommation d'aliments, et le taux de mortalité, ont été enregistrés sur une base hebdomadaire. De plus, des échantillons de sang ont été prélevés sur quatre oiseaux de chaque groupe pour déterminer les caractéristiques biochimiques sériques, et les titres d'anticorps contre NDV et IBV. Au cours de la semaine 5, quatre des poulets sélectionnés ont été euthanasiés, et les organes, coeur, foie, rate, bourse de Fabricius, ont été pesés. Des tissus de l'intestin, segments de 2 cm du duodénum, du jéjunum, de l'iléon et du cæcum, et de la bourse de Fabricius et thymus ont été prélevés pour des examens histologiques. Les résultats obtenus démontrent que les oiseaux dont l’alimentation était contaminée au DON présentaient une diminution (P < 0,05) du poids moyen aux jours 28 et 35 par rapport aux groupe témoin et groupes traités avec le SFA. De plus, l'inclusion de DON dans l'alimentation a réduit le titre d'anticorps contre le NDV (P < 0,05) et l'IBV (P < 0,001). Les paramètres biochimiques sériques, et le poids des organes pendant toute la période expérimentale (P > 0,05) n’ont pas montré de différences significatives. De plus, chez les oiseaux nourris avec des aliments contaminés, le DON a provoqué un processus de nécrose au niveau des villosités du duodénum et du thymus. Cependant, cela n'a pas modifié la morphométrie du tissu intestinal et de la bourse de Fabricius. Il a été conclu que les performances des poulets de chair, ainsi que la réponse vaccinale et paramètres histologiques, étaient négativement affectées par une alimentation contaminée par le DON. La supplémentation alimentaire avec un additif alimentaire à base de synbiotiques, serait une alternative pour atténuer les effets causés par cette mycotoxine chez les oiseaux. / Deoxynivalenol (DON) is one of the most abundant and important trichothecene mycotoxins produced by Fusarium species. Mycotoxin-contaminated feeds may negatively affect broiler chickens’ health, a sustainable approach to achieve mycotoxin elimination is necessary. An experiment was conducted to study the effects of deoxynivalenol (DON) on the performance of broilers, organ weights, serum biochemical parameters, immune response, and intestinal histology and to evaluate the efficacy of a commercial feed additive, based on synbiotics (SFA), with the ability to deepoxidize DON. In total, six hundred 1-d-old male broiler of a commercial strain (Ross 308) were vaccinated with combined live Newcastle disease virus (NDV) and infectious bronchitis virus (IBV) vaccine and were randomly allotted to 6 dietary treatments (100 birds in each group) for 5 wk. These groups were fed 3 diets naturally contaminated with DON at concentrations of < 0.5 (control diet), 1.5, and 3 mg/kg with and without SFA at 250 g/ton. The parameters including body weight, feed intake, and mortality rate were recorded on a weekly basis. In addition, weekly blood samples were collected from four birds in each group to determine serum biochemical characteristics and antibody titers to NDV and IBV. In week 5, four selected chickens were euthanized, and organs (heart, liver, spleen, bursa of Fabricius) were weighed. Tissues from intestine (2-cm segments of duodenum, jejunum, ileum, and caecum) and immune system (bursa of Fabricius and thymus) were collected for further histological examinations. The results indicated that DON-challenged birds had decreased (P < 0.05) average body weight (ABW) at days 28 and 35 as compared to control and treated groups with SFA. Furthermore, the inclusion of DON in the diet reduced antibody titer against NDV (P < 0.05), and IBV (P < 0.001). However, there were no significant differences in serum biochemical parameters and organs weight during the whole experimental period (P > 0.05). Moreover, in birds fed contaminated diets, DON caused necrosis in duodenum villus and in the thymus but did not alter the intestinal or the immune system morphometrics. It was concluded that broiler performance, histological, and immunological parameters were adversely affected by feeding diets contaminated with DON. However, the dietary supplementation with Synbiotics as a detoxifying agent would be an alternative to alleviate adverse effects on birds.

This Project identified the relationships between storage conditions, dry matter losses (DMLs) caused by Fusarium species in cereal grains and mycotoxin contamination and assessed novel control strategies for post-harvest grain management including chemical control and ozone. F. graminearum, F. verticillioides and F. langsethiae were inoculated on wheat, maize and oats and stored under environmental conditions where marginal to optimum spoilage and mycotoxin contamination can occur. DMLs were calculated from the CO2 produced and were significantly correlated with deoxynivalenol (DON), zearalenone (ZEA), fumonisins (FUMs) and T-2 and HT-2 toxins respectively. Mycotoxin levels in wheat and maize exceeded the EU legislative limits with 0.9-1% DMLs. Therefore, CO2 monitoring during storage can indicate the level of contamination in a stored batch. Using CO2 production data at different water activity (aw) and temperature conditions, the environmental regimes at which F. langsethiae can grow and contaminate oats with T-2 and HT-2 toxins were identified for the first time. Five acids were examined in vitro and little effect was observed on Fusarium growth, in the aqueous form, while the effect on mycotoxin production varied. Dissolved in ethanol, adipic, fumaric and ferulic acids inhibited fungal growth and controlled DON and FUMs, but T-2 toxin was stimulated by the ethanol. Two garlic essential oils, propyl-propylthiosulfinate (PTS) and propyl propylthiosulfonate (PTSO) were studied for the first time. In vitro, 200 ppm reduced fungal growth (50-100%) and mycotoxin production by >90%. The efficacy was species-dependent. In naturally contaminated oats of 0.93 aw stored for 20 days, 16 ppm PTSO reduced T-2 and HT-2 toxins by 66% and ochratoxin A (OTA) by 88%, while 200 ppm PTS reduced OTA by 95%. In wheat, 100 ppm PTS reduced DON and ZEA and 300 ppm PTS reduced fumonisins by 40-80%. PTSO:PTS (1:1) at 400 and 600 ppm was very effective against DON and ZEA in wheat of 0.92 aw. Ozone (O3) exposure at 200 ppm for 30 min delayed Fusarium spore germination on media of 0.98 aw and inhibited germination at 0.94 aw. O3 was more effective against fungal spores than mycelium and little effect was observed on growing cultures. In vitro, mycotoxin production after exposure depended on the stage of life of the fungi. O3 reduced fungal populations in grains. Mycotoxin production in wet grains treated with 100-200 ppm O3 for 60 min and stored for up to 30 days was reduced or completely inhibited, depending on the species and the exposure system. Simultaneous drying of the grain due to the O3 passage was observed.

No description available.