Abstract
Pistachio nuts are an important economic tree nut crop which is used directly or processed for many food-related activities. They can become colonized by mycotoxigenic spoilage fungi, especially Aspergillus flavus, mainly resulting in contamination with aflatoxins (AFs), especially aflatoxin B1 (AFB1). The prevailing climate in which these crops are grown changes as temperature and atmospheric CO2 levels increase, and episodes of extreme wet/dry cycles occur due to human industrial activity. The objectives of this study were to evaluate the effect of interacting Climate Change (CC)-related abiotic factors of temperature (35 vs. 37 °C), CO2 (400 vs. 1000 ppm), and water stress (0.98–0.93 water activity, aw) on (a) growth (b) aflD and aflR biosynthetic gene expression and (c) AFB1 production by two strains A. flavus (AB3, AB10) in vitro on milled pistachio-based media and when colonizing layers of shelled raw pistachio nuts. The A. flavus strains were resilient in terms of growth on pistachio-based media and the colonisation of pistachio nuts with no significant difference when exposed to the interacting three-way climate-related abiotic factors. However, in vitro studies showed that AFB1 production was significantly stimulated (p < 0.05), especially when exposed to 1000 ppm CO2 at 0.98–0.95 aw and 35 °C, and sometimes in the 37 °C treatment group at 0.98 aw. The relative expression of the structural aflD gene involved in AFB1 biosynthesis was decreased or only slightly increased, relative to the control conditions at elevated CO, regardless of the aw level examined. For the regulatory aflR gene expression, there was a significant (p < 0.05) increase in 1000 ppm CO2 and 37 °C for both strains, especially at 0.95 aw. The in situ colonization of pistachio nuts resulted in a significant (p < 0.05) stimulation of AFB1 production at 35 °C and 1000 ppm CO2 for both strains, especially at 0.98 aw. At 37 °C, AFB1 production was either decreased, in strain AB3, or remained similar, as in strain AB10, when exposed to 1000 ppm CO2. This suggests that CC factors may have a differential effect, depending on the interacting conditions of temperature, exposure to CO2 and the level of water stress on AFB1 production.
Highlights
Pistachio nuts (Pistacia vera L.) are one of the most economically important tree nut crops in countries such as the USA, Iran, Southern Europe and China
This required the development of an effective Hazard Analysis Critical Control Plan (HACCP) with appropriate Critical, Control Points in this chain focused on more efficient harvesting and post-harvest management of pistachios from Iran to meet the necessary legislative limits
Strains,growth growthon onthe thepistachio-based pistachio-basedmedium mediumwas wasrelatively relatively similar with no significant differences in relation to the three interacting abiotic condisimilar with no differences in relation to the three interacting abiotic conditions
Summary
Pistachio nuts (Pistacia vera L.) are one of the most economically important tree nut crops in countries such as the USA, Iran, Southern Europe and China. Poor drying and storage practices can exacerbate infection by A. flavus post-harvest [2,3]. Such infection results in significant contamination with aflatoxin B1 (AFB1 ), a class 1a carcinogen [4]. Iranian pistachio nuts were excluded from importation into the EU, because batches consistently exceeded the prevailing legislative limits for AFs [5]. This required the development of an effective Hazard Analysis Critical Control Plan (HACCP) with appropriate Critical, Control Points in this chain focused on more efficient harvesting and post-harvest management of pistachios from Iran to meet the necessary legislative limits
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