Miriam Haidukowski

Several Fusarium species have demonstrated the ability to thrive in saline soils and to tolerate or prefer high salt concentrations. In this context, the potential halophilic or halotolerant lifestyle of Fusarium pseudograminearum was investigated. Four isolates (3B, PVS-Fu 7, ColPat-1, and CBS 109956) were grown at different temperatures (10, 15, 20, 25, 30, and 35 °C) and NaCl concentrations (0, 7, 14, 21, and 28 g L⁻¹), and daily growth, mycotoxin production, and K⁺ and Na⁺ accumulation within hyphae were assessed. All F. pseudograminearum isolates exhibited strong adaptability to saline conditions, with significantly enhanced growth in the presence of NaCl. All isolates accumulated Na⁺ within their hyphae while retaining K⁺. The production of deoxynivalenol (DON) and zearalenone (ZEA) was generally suppressed following NaCl exposure, consistent with the known inhibitory effect of reduced water activity on mycotoxin biosynthesis. However, at 22 °C, two isolates, 3B and CBS 109956, showed no significant differences in ZEA production between the control (no salt) and the medium containing the lowest NaCl concentration tested (7 g L⁻¹). Notably, isolate 3B, obtained from the halophyte Salicornia europaea, retained the highest levels of both Na⁺ and K⁺ within hyphae and showed the greatest overall adaptation to salinity. These results confirm the hypothesis that the ability of F. pseudograminearum to colonize and infect a halophytic host is indicative of a halophilic lifestyle. In the context of increasing soil salinization, these findings help identify conditions that permit pathogen persistence without hazardous mycotoxin accumulation.

Alternaria species are important fungal pathogens occurring worldwide in wheat, causing both productive and qualitative losses, and posing a toxicological risk to human health due to the production of their mycotoxins in kernels. This study aimed to investigate the occurrence of Alternaria species and their mycotoxins in 48 wheat grain samples collected from the northeast to the southeast of Algeria. Seventy-two representative Alternaria strains were molecularly analyzed using a multi-locus sequence approach and evaluated for their capability to produce mycotoxins under in vitro conditions. Alternaria alternata, representing 42% of the strains, was the dominant species, followed to a lesser extent by species included in the Infectoriae section (26%). In addition, three species not previously reported in Algerian wheat, A. eureka, A. consortialis and A. tellustris, were identified, accounting for 5% of the total strains. Mycotoxin analyses showed high contamination of grains with alternariol monomethyl ether, alternariol and tenuazonic acid, occurring in 75, 69 and 35% of the samples, respectively. Moreover, 41 out of 48 samples showed the co-occurrence of multiple Alternaria mycotoxins. This study provides, for the first, time a clear picture of the occurrence and the distribution of Alternaria species on wheat in Algeria. Finally, the extensive monitoring activities carried out revealed the great biodiversity of Alternaria species able to colonize wheat grains. Moreover, findings on mycotoxin contamination raise concerns about the significant mycotoxigenic risk in Algerian wheat, emphasizing the need for strict monitoring and regulatory measures on Alternaria mycotoxins in food and feed.

Fusarium subglutinans and F. temperatum are widely distributed maize pathogens recognized as distinct species with a species-specific chemotype based on patterns of mycotoxins. Recent comparative genomic analysis revealed that genomes of both species carry a complete beauvericin (Bea) biosynthetic genes cluster, but the key gene Bea1 in F. subglutinans is not functional likely due to a large insertion (NRPS22ins) and multiple mutations (SNP298 and SNP528). We used the recently published genome sequences for these species to develop PCR markers for investigating the distribution of three main mutations in the Bea1 gene in a large collection of strains of both species from around the world. We also designed a PCR assay for a rapid and reliable discrimination of both species in the evaluation of crop exposure to mycotoxins. Overall, our results showed that SNP528 was the most common mutation, followed by NRPS22ins and SNP298. Moreover, phylogenetic analyses suggest that non-synonymous SNPs have occurred first, and that the resulting inactivation of BEA production has caused the accumulation of other polymorphisms, including the NRPS22ins, in the entire gene-coding region. The screening for genetic differences between these species could guide future crop management strategies.

Aflatoxins (AFs) are toxic secondary metabolites produced by Aspergillus spp. and are found in food and feed as contaminants worldwide. Due to climate change, AFs occurrence is expected to increase also in western Europe. Therefore, to ensure food and feed safety, it is mandatory to develop green technologies for AFs reduction in contaminated matrices. With this regard, enzymatic degradation is an effective and environmentally friendly approach under mild operational conditions and with minor impact on the food and feed matrix. In this work, Ery4 laccase, acetosyringone, ascorbic acid, and dehydroascorbic acid were investigated in vitro, then applied in artificially contaminated corn for AFB₁ reduction. AFB₁ (0.1 µg/mL) was completely removed in vitro and reduced by 26% in corn. Several degradation products were detected in vitro by UHPLC-HRMS and likely corresponded to AFQ₁, epi-AFQ₁, AFB₁-diol, or AFB₁dialehyde, AFB_2a, and AFM₁. Protein content was not altered by the enzymatic treatment, while slightly higher levels of lipid peroxidation and H₂O₂ were detected. Although further studies are needed to improve AFB₁ reduction and reduce the impact of this treatment in corn, the results of this study are promising and suggest that Ery4 laccase can be effectively applied for the reduction in AFB₁ in corn.