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Aflatoxins (AFs) are toxic fungal secondary metabolites that are commonly detected in food commodities. Currently, there is a lack of generic methods capable of determining AFs both at postharvest stages in agricultural products and preharvest stages, namely, the agricultural soil. Here, we present a simple and reliable method for quantitative analysis of AFs in soil and food matrices at environmentally relevant concentrations for the first time, using the same extraction procedure and chromatography, either by HPLC-FLD or LC–MS. AFs were extracted from matrices by ultrasonication using an acetonitrile/water mixture (84:16, v + v) without extensive and time-consuming cleanup procedures. Food extracts were defatted with n-hexane. Matrix effects in terms of signal suppression/enhancement (SSE) for HPLC-FLD were within ±20% for all matrices tested. For LC–MS, the SSE values were mostly within ±20% for soil matrices but outside ±20% for all food matrices. The sensitivity of the method allowed quantitative analysis even at trace levels with quantification limits (LOQs) between 0.04 and 0.23 μg kg–1 for HPLC-FLD and 0.06–0.23 μg kg–1 for LC–MS. The recoveries ranged from 64 to 92, 74 to 101, and 78 to 103% for fortification levels of 0.5, 5, and 20 μg kg–1, respectively, with repeatability values of 2–18%. The validation results are in accordance with the quality criteria and limits for mycotoxins set by the European Commission, thus confirming a satisfactory performance of the analytical method. Although reliable analysis is possible with both instruments, the HPLC-FLD method may be more suitable for routine analysis because it does not require consideration of the matrix.

While mycotoxins are generally regarded as food contamination issues, there is growing interest in mycotoxins as environmental pollutants. The main sources of trichothecene and zearalenone mycotoxins in the environment are mainly attributed to Fusarium infested fields, where mycotoxins can wash off in infested plants or harvest residues. Subsequently, mycotoxins inevitably enter the soil. In this context, investigations into the effects, fate, and transport are still needed. However, there is a lack of analytical methods used to determine Fusarium toxins in soil matrices. We aimed to validate an analytical method capable of determining the toxins nivalenol (NIV), deoxynivalenol (DON), 15-acetyl-deoxynivalenol (15-AcDON), and zearalenone (ZEN), at environmentally relevant concentrations, in five contrasting agricultural soils. Soils were spiked at three levels (3, 9 and 15ng/g), extracted by solid-liquid extraction assisted with ultrasonication, using a generic solvent composition of acetonitrile:water 84:16 (v:v) and measured by LC–HRMS. Method validation was successful for NIV, DON, and 15-AcDON with mean recoveries > 93% and RSDr < 10%. ZEN failed the validation criteria. The validated method was applied to eight conventionally managed maize field soils during harvest season, to provide a first insight into DON, NIV, and 15-AcDON levels. Mycotoxins were present in two out of eight sampled maize fields. Soil mycotoxin concentrations ranged from 0.53ng/g–19.4ng/g and 0.8ng/g–2.2ng/g for DON and NIV, respectively. Additionally, we found indication that “hot-spot” concentrations were restricted to small scales ( <5cm) with implications for field scale soil monitoring strategies.

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Aflatoxins (AFs) are fungal secondary metabolites frequently detected in soil that exhibit in vitro toxicity to certain soil microorganisms. However, microbial responses at different levels and in complex systems such as the soil environment have not been systematically studied. Therefore, we investigated multiple microbial responses in two different soils (sandy loam and clay) to aflatoxin B1 (AFB1) at environmentally relevant concentrations (0.5–500 µg kg−1) during a 28-day incubation. General microbial parameters for biomass (microbial biomass carbon and ergosterol), activity (glucose-induced and basal respiration), and catabolic functionality (substrate utilization patterns) were assessed. We observed minor and transient effects in both soils. In sandy loam, we found negative effects on activity and catabolic functionality with increased metabolic quotient, while clay soil exhibited stimulation for the same parameters, suggesting a hormetic effect due to reduced bioavailability through sorption onto clay minerals. Our results indicate that AFB1 does not pose a threat to general microbial indicators under the test conditions in soils without previous AF contamination. Given the toxic potential of AFs to specific microorganisms, further studies should investigate responses at higher taxonomic and functional levels in natural environments of aflatoxigenic fungi, such as tropical soils, and including additional physicochemical stressors.

In a 28-days experiment, we investigated the dissipation of aflatoxin B1 (AFB1) (0.5-500 μg kg−1) by microbial (MD) and photodegradation (PD) in two contrasting soils (sandy loam and clay). Sterile incubation in darkness served as control (C). AFB1 was degraded in all scenarios according to simple first-order kinetics with 50% dissipation times of 20 - 32 (PD), 19 - 48 (MD), and 56 - 65 days (C), respectively. Dissipation rates were significantly lower (p<0.001) in the clay soil, likely due to photoquenching and strong binding of AFB1 by clay minerals and humic substances. In the sandy loam, dissipation rate of MD decreased in function of initial AFB1 concentration, probably due to toxic effects on degrading microbes. In contrast, in the clay soil the dissipation rate increased with increasing concentration up to 250 μg kg−1, followed by a sharp decrease at 500 μg kg−1, indicating an effect of soil texture on the bioavailability of AFB1 to soil microbes. AFB2a was identified as a transformation product in all scenarios. These results confirm the function of soil for AFB1 degradation, which is modulated by abiotic and biotic processes, soil characteristics and initial AFB1 concentration.

In a 28-days experiment, we investigated the dissipation of aflatoxin B1 (AFB1) (0.5–500 $${\upmu }\text {g}\,\text {kg}^{-1}$$ μ g kg - 1 ) by microbial (MD) and photodegradation (PD) in two contrasting soils (sandy loam and clay). Sterile incubation in darkness served as control (C). AFB1 was degraded in all scenarios according to simple first-order kinetics with 50% dissipation times of 20–32 (PD), 19–48 (MD), and 56–65 days (C), respectively. Dissipation rates were significantly lower ($$\text {p}<0.001$$ p < 0.001 ) in the clay soil than in the sandy loam soil, likely due to photoquenching and strong binding of AFB1 by clay minerals and humic substances. In the sandy loam, dissipation rate of MD decreased in function of initial AFB1 concentration, probably due to toxic effects on degrading microbes. In contrast, in the clay soil the dissipation rate increased with increasing concentration up to 250 $$\upmu \text {g}\,\text {kg}^{-1}$$ μ g kg - 1 , followed by a sharp decrease at 500 $$\upmu \text {g}\,\text {kg}^{-1}$$ μ g kg - 1 , indicating an effect of soil texture on the bioavailability of AFB1 to soil microbes. AFB2a was identified as a transformation product in all scenarios. These results confirm the function of soil for AFB1 degradation, which is modulated by abiotic and biotic processes, soil characteristics and initial AFB1 concentration.