Abstract
This study aimed to assess the occurrence of toxigenic fungi and mycotoxin contamination in stored wheat grains by using advanced molecular and analytical techniques. A multiplex polymerase chain reaction (PCR) strategy was established for rapid identification of mycotoxigenic fungi, and an improved analytical method was developed for simultaneous multi-mycotoxin determination in wheat grains by liquid chromatography-tandem mass spectrometry (LC/MS/MS) without the need for any clean-up. The optimized multiplex PCR method was highly specific in detecting fungal species containing species-specific and mycotoxin metabolic pathway genes. The method was applied for evaluation of 34 wheat grain samples collected from storage warehouses for the presence of mycotoxin-producing fungi, and a few samples were found positive for Fusarium and Aspergillus species. Further chemical analysis revealed that 17 samples contained mycotoxins above the level of detection, but only six samples were found to be contaminated over the EU regulatory limits with at least one mycotoxin. Aflatoxin B1, fumonisins, and deoxynivalenol were the most common toxins found in these samples. The results showed a strong correlation between the presence of mycotoxin biosynthesis genes as analyzed by multiplex PCR and mycotoxin detection by LC/MS/MS. The present findings indicate that a combined approach might provide rapid, accurate, and sensitive detection of mycotoxigenic species and mycotoxins in wheat grains.
Highlights
Wheat grain and associated by-products are important sources of energy and protein for humans and all classes of farm animals
When grains are colonized by moulds there is a significant risk of contamination with mycotoxins, which are toxic chemical products, formed as secondary metabolites by these fungi
Toxigenic fungi in crops have been divided into two distinct classes: ‘field fungi’, which invade and produce their toxins before harvest; and ‘storage’ fungi, which become a problem after harvest
Summary
Wheat grain and associated by-products are important sources of energy and protein for humans and all classes of farm animals. When grains are colonized by moulds there is a significant risk of contamination with mycotoxins, which are toxic chemical products, formed as secondary metabolites by these fungi. Many species of Fusarium, Aspergillus, Penicillium, and Alternaria are recognised as plant pathogens but are sources of important mycotoxins of concern in relation to animal and human health [1]. Toxigenic fungi in crops have been divided into two distinct classes: ‘field fungi’ (or plant pathogens), which invade and produce their toxins before harvest; and ‘storage’ (or saprophytic) fungi, which become a problem after harvest. Some fungi might belong to both classes and colonize grains before and after harvest.
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