Interacting Abiotic Factors Affect Growth and Mycotoxin Production Profiles of Alternaria Section Alternaria Strains on Chickpea-Based Media.

Chickpea is susceptible to fungal infection and mycotoxin contamination. Argentina exports most of its chickpea production; thus, its quality is of concern. The Alternaria fungal genus was found to be prevalent in chickpea samples from Argentina. The species within this genus are able to produce mycotoxins, such as alternariol (AOH), alternariol monomethyl ether (AME), and tenuazonic acid (TA). In this context, we evaluated the effect of water activity (0.99, 0.98, 0.96, 0.95, 0.94, 0.92, and 0.90 aW), temperature (4, 15, 25, and 30 °C), incubation time (7, 14, 21, and 28 days), and their interactions on mycelial growth and AOH, AME, and TA production on chickpea-based medium by two A. alternata strains and one A. arborescens strain isolated from chickpea in Argentina. Maximum growth rates were obtained at the highest aW (0.99) and 25 °C, with growth decreasing as the aW of the medium and the temperature were reduced. A. arborescens grew significantly faster than A. alternata. Mycotoxin production was affected by both variables (aW and temperature), and the pattern obtained was dependent on the strains/species evaluated. In general, both A. alternata strains produced maximum amounts of AOH and AME at 30 °C and 0.99-0.98 aW, while for TA production, both strains behaved completely differently (maximum levels at 25 °C and 0.96 aW for one strain and 30 °C and 0.98 aW for the other). A. arborescens produced maximum amounts of the three toxins at 25 °C and 0.98 aW. Temperature and aW conditions for mycotoxin production were slightly narrower than those for growth. Temperature and aW conditions assayed are those found during chickpea grain development in the field, and also could be present during storage. This study provides useful data on the conditions representing a risk for contamination of chickpea by Alternaria toxins.

Fusarium chaquense, sp. nov, a novel type A trichothecene-producing species from native grasses in a wetland ecosystem in Argentina.

The Chaco wetland is among the most biologically diverse regions in Argentina. In collections of fungi from asymptomatic native grasses (Poaceae) from the wetlands, we identified isolates of Fusarium that were morphologically similar to F. armeniacum, but distinct from it by their production of abundant microconidia. All the isolates had identical, or nearly identical, partial sequences of TEF1 and RPB2. But they were distinct from reference sequences from F. armeniacum and Fusarium species closely related to it. Phylogenetic analysis of 34 full-length housekeeping gene sequences retrieved from whole genome sequences of three Chaco wetland isolates, 29 genes resolved the isolates as an exclusive clade within the F. sambucinum species complex. Based on results of the morphological and phylogenetic analysis, we concluded that the Chaco wetland isolates are a distinct and novel species, herein described as Fusarium chaquense, sp. nov., which is closely related to F. armeniacum. F. chaquense in culture can produce the trichothecenes T-2 and HT-2 toxin, neosolaniol, diacetoxyscirpenol, and monoacetoxyscirpenol, as well as beauvericin and the pigment aurofusarin. Genome sequence analysis also revealed the presence of three previously described loci required for trichothecene biosynthesis. This research represents the first study of Fusarium in a natural ecosystem in Argentina.

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.

Fumonisin occurrence in wheat-based products from Argentina.

In Argentina, wheat is the most consumed cereal by the human population. Since fumonisins occurence in wheat grains and wheat-based products have been reported worldwide, a survey was conducted in order to determine fumonisin contamination in 91 wheat-based products (white wheat flour samples, wheat flour used at bakery products and whole-wheat flour samples) collected from different retail stores of Rio Cuarto city in Argentina using HPLC-MS/MS. Sixty-seven samples (74%) showed contamination by fumonisins. From these samples, 16 showed fumonisin levels between LOD and LOQ (between 0.01 to 0.05 ng/g), while fumonisins (FB1 + FB2) in quantifiable samples ranged from 0.05 ng/g to 18.9 ng/g. Although FB1 was more prevalent, FB2 was foun3d in higher levels than FB1. Overall, fumonisin prevalence was high, but concentrations were far below EU or USA limits set for maize and maize-based products.

Preliminary Study on the Use of Chitosan as an Eco-Friendly Alternative to Control Fusarium Growth and Mycotoxin Production on Maize and Wheat.

The objectives of the present study were to determine the combined effects of chitosan and water activity (aW) on growth and mycotoxin production in situ on the two most important Fusarium species (F. proliferatum and F. verticillioides) present on maize, and on F. graminearum, the main pathogen causing Fusarium head blight on wheat. Results showed that low-molecular-weight chitosan with more than 70% deacetylation at the lowest dose used (0.5 mg/g) was able to reduce deoxynivalenol (DON) and fumonisin (FBs) production on irradiated maize and wheat grains. Growth rates of F. graminearum also decreased at the lowest chitosan dose used (0.5 mg/g), while F. verticillioides and F. proliferatum growth rates were reduced at 0.98 aW at the highest chitosan dose used (2 mg/g). Since mycotoxins are unavoidable contaminants in food and feed chains, their presence needs to be reduced in order to minimize their effects on human and animal health and to diminish the annual market loss through rejected maize and wheat; in this scenario, pre- and post-harvest use of chitosan could be an important alternative.

Presence of Multiple Mycotoxins and Other Fungal Metabolites in Native Grasses from a Wetland Ecosystem in Argentina Intended for Grazing Cattle.

The aim of this study was to evaluate the occurrence of several fungal metabolites, including mycotoxins in natural grasses (Poaceae) intended for grazing cattle. A total number of 72 and 77 different metabolites were detected on 106 and 69 grass samples collected during 2011 and 2014, respectively. A total of 60 metabolites were found across both years. Among the few mycotoxins considered toxic for ruminants, no samples of natural grasses were contaminated with aflatoxins, ochratoxin A, ergot alkaloids, and gliotoxin, among others. However, we were able to detect important metabolites (toxic to ruminants) such as type A trichothecenes, mainly T-2 toxin and HT-2 toxin (up to 5000 µg/kg each), and zearalenone (up to 2000 µg/kg), all at very high frequencies and levels. Other fungal metabolites that were found to be prevalent were other Fusarium metabolites like beauvericin, equisetin and aurofusarin, metabolites produced by Alternaria spp., sterigmatocystin and its precursors and anthrachinone derivatives. It is important to point out that the profile of common metabolites was shared during both years of sampling, and also that the occurrence of important metabolites is not a sporadic event. Considering that this area of temperate grassland is used for grazing cattle all year long due to the richness in palatable grasses (Poaceae), the present work represents a starting point for further studies on the occurrence of multi-mycotoxins in natural grasses in order to have a complete picture of the extent of cattle exposure. Also, the present study shows that the presence of zeranol in urine of beef cattle may not be a consequence of illegal use of this banned substance, but the product of the natural occurrence of zearalenone and α-zearalenol in natural grasses intended for cattle feeding.