Impacts of Temperature and Water Activity Interactions on Growth, Aflatoxin B1 Production and Expression of Major Biosynthetic Genes of AFB1 in Aspergillus flavus Isolates

The contamination of peanuts, with Aspergillus flavus and subsequent aflatoxins (AFs) is considered to be one of the most serious, safety problems in the world. Water activity (a(w)) and temperature are limiting, factors for fungal growth and aflatoxin production during storage. The objectives of this study were to integrate data on the effects of temperature (34, 37, and 42 °C) and water activity (a(w); 0.85, 0.90, and 0.95) on growth rate aflatoxin B1 (AFB1) production and up- or-downregulation of the molecular expression of biosynthetic AFB1 genes divided into three types based on their A. flavus isolate composition and AFB1 capacity in vitro: A. flavus KSU114 (high producer), A. flavus KSU114 (low producer), and A. flavus KSU121 (non-producer). The A. flavus isolates were shown to be resilient in terms of growth on yeast extract sucrose agar media when exposed to temperature and water activity as pivotal environmental factors. The optimal conditions for the fungal growth of three isolates were a temperature of 34 °C and water activity of 0.95 a(w); there was very slow fungal growth at the highest temperature of 42 °C, with different a(w) values causing inhibited fungal growth. The AFB1 production for the three isolates followed the same pattern with one exception: A. flavus KSU114 failed to produce any AFB1 at 42 °C with different a(w) values. All tested genes of A. flavus were significantly up- or downregulated under three levels of interaction between temperature and a(w). The late structural genes of the pathway were significantly upregulated at 34 °C under a(w) 0.95, although aflR, aflS and most of the early structural genes were upregulated. Compared to 34 °C with an a(w) value of 0.95, most of the expressed genes were significantly downregulated at 37 and 42 °C with a(w) values of 0.85 and 0.90. Additionally, two regulatory genes were downregulated under the same conditions. The expression level of laeA was also completely associated with AFB1 production, while the expression level of brlA was linked to A. flavus colonization. This information is required to forecast the actual effects of climate change on A. flavus. The findings can be applied to improve specific food technology processes and create prevention strategies to limit the concentrations of potential carcinogenic substances in peanuts and their derivatives.