Type A Trichothecene Metabolic Profile Differentiation, Mechanisms, Biosynthetic Pathways, and Evolution in Fusarium Species-A Mini Review.

Trichothecenes are the most common Fusarium toxins detected in grains and related products. Type A trichothecenes are among the mycotoxins of greatest concern to food and feed safety due to their high toxicity. Recently, two different trichothecene genotypes within Fusarium species were reported. The available information showed that Tri1 and Tri16 genes are the key determinants of the trichothecene profiles of T-2 and DAS genotypes. In this review, polymorphisms in the Tri1 and Tri16 genes in the two genotypes were investigated. Meanwhile, the functions of genes involved in DAS and NEO biosynthesis are discussed. The possible biosynthetic pathways of DAS and NEO are proposed in this review, which will facilitate the understanding of the synthesis process of trichothecenes in Fusarium strains and may also inspire researchers to design and conduct further research. Together, the review provides insight into trichothecene profile differentiation and Tri gene evolutionary processes responsible for the structural diversification of trichothecene produced by Fusarium.

Intestinal toxicity of the new type A trichothecenes, NX and 3ANX.

NX and its acetylated form 3ANX are two new type A trichothecenes produced by Fusarium graminearum whose toxicity is poorly documented. The aim of this study was to obtain a general view of the intestinal toxicity of these toxins. Deoxynivalenol (DON), which differs from NX by the keto group at C8, served as a benchmark. The viability of human intestinal Caco-2 cells decreased after 24 h of exposure to 3 µM NX (-21.4%), 3 µM DON (-20.2%) or 10 µM 3ANX (-17.4%). Histological observations of porcine jejunal explants exposed for 4 h to 10 µM of the different toxins showed interstitial edema and cellular debris. Explants exposed to NX also displayed cell vacuolization, a broken epithelial barrier and high loss of villi. Whole transcriptome profiling revealed that NX, DON and 3ANX modulated 369, 146 and 55 genes, respectively. Functional analyses indicated that the three toxins regulate the same gene networks and signaling pathways mainly; cell proliferation, differentiation, apoptosis and growth, and particularly immune and pro-inflammatory responses. Greater transcriptional impacts were observed with NX than with DON. In conclusion, our data revealed that the three toxins have similar impacts on the intestine but of different magnitude: NX > DON ≫ 3ANX. NX and 3ANX should consequently be included in overall risk analysis linked to the presence of trichothecenes in our diet.

Ecophysiology of Fusarium chaquense a Novel Type A Trichothecene Producer Species Isolated from Natural Grasses.

Fusarium chaquense, a recently formally described novel species, has been identified as an T-2 toxin (T-2), HT-2 toxin (HT-2) and other toxins producer in natural grasses (Poaceae) from Argentina. The major objective of this study was to describe the effect of water activity (aW, 0.995, 0.98, 0.95, 0.93 and 0.91), temperature (15, 25 and 30 °C) and incubation time (5, 15 and 25 days) on growth and to evaluate the production of T-2, HT-2 toxins and beauvericin (BEA) by two F. chaquense strains in a grass-based media. The results showed a wide range of conditions for F. chaquense growth and mycotoxin production. Both strains had a maximum growth rate at the highest aW (0.995) and 25 °C. Regarding mycotoxin production, more T-2 than the other analysed mycotoxins were produced by the two strains. T-2 production was favoured at 0.995 aW and 30 °C, while HT-2 production at 0.98-0.95 aW and 15 °C. The maximum levels of BEA were produced at 0.995 aW and 25-30 °C. Two-dimensional profiles of aW by temperature interactions were obtained from these data in order to identify areas where conditions indicate a significant risk of mycotoxins accumulation on grass. For its versatility on growth and mycotoxin production in a wide range of aW and temperatures, F. chaquense would have an adaptive advantage over other Fusarium species, and this would explain its high frequency of isolation in natural grasses grown up in the Chaco wetlands.

Development of isotope dilution-liquid chromatography/tandem mass spectrometry for the accurate determination of type-A trichothecenes in grains.

We report the development of a highly accurate method based on isotope dilution liquid chromatography-tandem mass spectrometry (ID-LC-MS/MS) for the simultaneous determination of four major type-A trichothecenes in cereal grains. Uniformly labeled [13C] analogues of the target analytes were used as internal standards. An expedient sample preparation procedure was established. After extraction with acetonitrile/water (84:16; v/v), further clean-up was performed using MycoSep 227 solid-phase extraction cartridges. Unlike the commonly used immunoaffinity columns having strong selectivity for only certain target analytes, the cartridges allowed the simultaneous recovery of all four mycotoxins and efficient elimination of co-extracted matrix interferences. The ID-LC-MS/MS method exhibited very good analytical performance in the concentration range of 10-200 µg/kg; accuracy ranged from 97 to 103% with intra-day and inter-day relative standard deviations of less than 5% and 4%, respectively. Measurement uncertainties were generally below 5%. The applicability of the method was assessed by measuring the target mycotoxins in several samples at sub-µg/kg levels.

Genetic dereplication of Trichoderma hypoxylon reveals two novel polycyclic lactones.

Previous study demonstrated large scale production of trichochecenes which limited the discovery of novel metabolites in Trichoderma hypoxylon. By genetic deletion of trichothecene synthase encoding gene thtri5, we created the dereplication mutant which eliminated the production of trichothecenes. Through chemical isolation, we characterized a couple of rare new polycyclic lactones tricholactones A and B from the thtri5 deletion strain. The structures of these two compounds were well determined by NMR, HR-ESI-MS and IECD analysis.