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International audience ; This study examined the relationship between storage environmental factors (water activity (aw) (0.89-0.97) and temperature (15-30ºC)), colonisation of wheat and maize by Fusarium graminearum and F. verticillioides respectively and the dry matter losses (DMLs) caused, and quantified the contamination with deoxynivalenol (DON), zearalenone (ZEA) and fumonisins (FUMs) during storage. Fungal growth was assessed by the amount of CO2 produced under different interacting conditions of aw and temperature. DMLs were quantified using the cumulative CO2 data and these were shown to increase as temperature and aw increased. The amount of DON, ZEA (wheat for human consumption) and FUMs (feed maize) produced was significantly affected by the storage conditions. The three toxins however showed different patterns of production. Optimum for DON was at the wettest conditions (0.97 aw) and the highest temperature assessed (30ºC), while for ZEA this shifted to 25ºC. FUMs were produced in higher amounts in maize at 30ºC and 0.97 aw, however, at intermediate aw levels (0.955 aw) the highest production occurred at 25ºC followed by 20ºC. Polynomial models were developed for the effect of the storage factors on DMLs and toxin production. DMLs under different environmental conditions were significantly correlated with DON and FUMs. DON contamination was above the EU limits in at least 80% of the wheat samples with DMLs>1%, while at least 70% of the same samples contained ZEA above the respective EU legislative limits. Similarly, at least 75% of the maize samples with DMLs≥0.9% exceeded the EU limits for the sum of fumonisins in feed. These results show that it may be possible to use temporal CO2 production during storage of grains as an indicator of the level of contamination of the grain with mycotoxins.

Chillies are an important spice in many regions of the world. During the production and processing phases, they are prone to infection by mycotoxigenic fungi, especially Aspergillus Section Flavi species and contamination with aflatoxins. There is significant interest in controlling aflatoxin B1 (AFB1) contamination in such spices to ensure they remain below the legislative limits for human consumption. This study initially examined the potential efficacy of sodium metabisulphite (NaMBS, up to 5000 mg/L) for the control of growth and AFB1 production by Aspergillus flavus strains in vitro on a chilli-based medium modified to 0.93–0.995 water activity, aw). Subsequent studies examined efficacy of NaMBS in stored naturally contaminated chilli powder and whole red chillies, or when inoculated with A. flavus at 0.70, 0.80, 0.90, 0.95 aw and 30 °C for 20 days. NaMBS was effective in vitro in controlling growth of the three strains of A. flavus at the different aw levels examined. No growth occurred with >500–1250 NaMBS mg/L. AFB1 was only produced at 0.98 aw with complete inhibition at ≥1250 mg/L of the preservative at 30 °C. The in vitro ED50, ED90 for NaMBS varied with aw conditions. In situ studies with chilli powder and whole red dry chillies (naturally contaminated or + conidial inoculum of A. flavus) showed that at 0.70 and 0.80 aw, NaMBS treatments of 1000 and 2000 mg/L resulted in AFB1 contamination levels below the legislative limits for spices. However, under wetter conditions at 0.90 and 0.95 aw, AFB1 contamination of stored chilli powder, −/+ A. flavus inoculum, were above the legislative limits, even with 2000 mg/L NaMBS and 20 days storage at 30 °C. Stored whole chillies inoculated with A. flavus resulted in a reduction of populations, especially at 2000 mg/L NaMBS. Control of AFB1 was achieved only with 2000 mg/L NaMBS treatment after 10 and 20 days storage. Studies with commercial laminated sheets containing immobilised NaMBS with slow release of SO2 properties significantly reduced fungal populations and effectively controlled AFB1 contamination of the stored and packaged whole red chillies.

The objective of this study was to compare the effect of different storage moisture conditions (0.70, 0.85, 0.90 and 0.95 water activity, aw) and temperatures (20, 25, 30 °C) on (a) respiration rates (R) and dry matter loss (DML) of paddy and brown rice and (b) quantify aflatoxin B1 (AFB1) production by isolates of Aspergillus flavus from the rice samples and (c) inoculation of both rice types with A. flavus under these storage conditions on R, DML and AFB1 contamination. There was an increase in temporal CO2 production with wetter and warmer conditions in naturally contaminated rice. Higher R and consequently, % DML, were generally found in the brown rice (21%) while in paddy rice this was only up to 3.5% DML. From both rice types, 15 (83.3%) of 18 A. flavus isolates produced detectable levels of AFB1 in a range 2.5–1979.6 μg/kg. There was an increase in DML in both rice types inoculated with A. flavus as temperature and aw were increased. Interestingly very little AFB1 was detected in paddy rice, but significant contamination occurred in the brown rice. The %DML in the control and A. flavus inoculated rice increased with temperature and aw at both 25 and 30 °C from 1-2% to 15–20% DML at 30 °C and 0.95 aw. All the inoculated rice samples had AFB1 levels above the EU legislative limits for contamination in other temperate cereals and products derived from cereals (=2 μg/kg). Even samples with % DML as low as 0.2% had AFB1 contamination levels twice the limits for other cereals. These results suggest that the mycotoxin contamination risk in staple commodities like rice, is influenced by whether the rice is processed or not, and that measurement of R rates can be used to predict the relative risk of AFB1 contamination in such staple commodities.

Batches of grain are accepted or rejected based on average mycotoxin concentrations in a composite grain sample. Spatial analysis of mycotoxins in two grain bulks was performed to determine the spatial distribution of toxins, whether they were co-located and the proportions of grain over legislative limits. The 2D distribution of deoxynivalenol (DON) and ochratoxin A (OTA) in a truck load of wheat grain was analysed, as was the distribution of fumonisins (FB1 and FB2) in a 3D maize grain pile. The data had been previously analysed, but results here show that highly skewed data would need to be transformed to investigate spatial autocorrelation properly. In the truck of wheat grain, DON and OTA showed co-variation and, in contrast to previous studies, OTA showed spatial structure when converted to normal scores. Spatial analysis of the maize pile showed that FB1 and FB2 contamination levels were each highest near the outer face and base of the grain pile. Simulations for both grain bulks showed that, for average toxin concentrations close to legislative limits, the proportion of grain over the legislative limits can vary greatly and could be very small when toxin contamination is highly positively skewed. The implications of the results for management were considered. Post-harvest, strategically placed sensors could be used to monitor environmental conditions within the stored grain in real time and detect the first signs of spoilage allowing swift remediative action so less grain is wasted. Pre-harvest approaches for mycotoxin management are suggested as additional food waste reduction strategies.

Two garlic-derived compounds, Propyl Propane Thiosulfonate (PTS) and Propyl Propane Thiosulfinate (PTSO), were examined for their efficacy against mycotoxigenic Fusarium species (F. graminearum, F. langsethiae, F. verticillioides). The objectives were to assess the inhibitory effect of these compounds on growth and mycotoxin production in vitro, and in situ in artificially inoculated wheat, oats and maize with one isolate of each respectively, at different water activity (aw) conditions when stored for up to 20 days at 25 °C. In vitro, 200 ppm of either PTS or PTSO reduced fungal growth by 50–100% and mycotoxin production by >90% depending on species, mycotoxin and aw conditions on milled wheat, oats and maize respectively. PTS was generally more effective than PTSO. Deoxynivalenol (DON) and zearalenone (ZEN) were decreased by 50% with 80 ppm PTSO. One-hundred ppm of PTS reduced DON and ZEN production in wheat stored at 0.93 aw for 20 days, although contamination was still above the legislative limits. Contrasting effects on T-2/HT-2 toxin contamination of oats was found depending on aw, with PTS stimulating production under marginal conditions (0.93 aw), but at 0.95 aw effective control was achieved with 100 ppm. Treatment of stored maize inoculated with F. verticilliodies resulted in a stimulation of total fumonsins in most treatments. The potential use of such compounds for mycotoxin control in stored commodities is discussed.

Two garlic-derived compounds, Propyl Propane Thiosulfonate (PTS) and Propyl Propane Thiosulfinate (PTSO), were examined for their efficacy against mycotoxigenic Fusarium species (F. graminearum, F. langsethiae, F. verticillioides). The objectives were to assess the inhibitory effect of these compounds on growth and mycotoxin production in vitro, and in situ in artificially inoculated wheat, oats and maize with one isolate of each respectively, at different water activity (a(w)) conditions when stored for up to 20 days at 25 °C. In vitro, 200 ppm of either PTS or PTSO reduced fungal growth by 50–100% and mycotoxin production by >90% depending on species, mycotoxin and a(w) conditions on milled wheat, oats and maize respectively. PTS was generally more effective than PTSO. Deoxynivalenol (DON) and zearalenone (ZEN) were decreased by 50% with 80 ppm PTSO. One-hundred ppm of PTS reduced DON and ZEN production in wheat stored at 0.93 a(w) for 20 days, although contamination was still above the legislative limits. Contrasting effects on T-2/HT-2 toxin contamination of oats was found depending on a(w), with PTS stimulating production under marginal conditions (0.93 a(w)), but at 0.95 a(w) effective control was achieved with 100 ppm. Treatment of stored maize inoculated with F. verticilliodies resulted in a stimulation of total fumonsins in most treatments. The potential use of such compounds for mycotoxin control in stored commodities is discussed.

Zearalenone (ZEN) contamination from Fusarium graminearum colonization is particularly important in food and feed wheat, especially during post-harvest storage with legislative limits for both food and feed grain. Indicators of the relative risk from exceeding these limits would be useful. We examined the effect of different water activities (aw; 0.95–0.90) and temperature (10–25 °C) in naturally contaminated and irradiated wheat grain, both inoculated with F. graminearum and stored for 15 days on (a) respiration rate; (b) dry matter losses (DML); (c) ZEN production and (d) relationship between DML and ZEN contamination relative to the EU legislative limits. Gas Chromatography was used to measure the temporal respiration rates and the total accumulated CO2 production. There was an increase in temporal CO2 production rates in wetter and warmer conditions in all treatments, with the highest respiration in the 25 °C × 0.95 aw treatments + F. graminearum inoculation. This was reflected in the total accumulated CO2 in the treatments. The maximum DMLs were in the 0.95 aw/20–25 °C treatments and at 10 °C/0.95 aw. The DMLs were modelled to produce contour maps of the environmental conditions resulting in maximum/minimum losses. Contamination with ZEN/ZEN-related compounds were quantified. Maximum production was at 25 °C/0.95–0.93 aw and 20 °C/0.95 aw. ZEN contamination levels plotted against DMLs for all the treatments showed that at ca. 1.0% DML, the risk was high. This type of data is important in building a database for the development of a post-harvest decision support system for relative risks of different mycotoxins.

Zearalenone (ZEN) contamination from Fusarium graminearum colonization is particularly important in food and feed wheat, especially during post-harvest storage with legislative limits for both food and feed grain. Indicators of the relative risk from exceeding these limits would be useful. We examined the effect of different water activities (aw; 0.95–0.90) and temperature (10–25 °C) in naturally contaminated and irradiated wheat grain, both inoculated with F. graminearum and stored for 15 days on (a) respiration rate; (b) dry matter losses (DML); (c) ZEN production and (d) relationship between DML and ZEN contamination relative to the EU legislative limits. Gas Chromatography was used to measure the temporal respiration rates and the total accumulated CO2 production. There was an increase in temporal CO2 production rates in wetter and warmer conditions in all treatments, with the highest respiration in the 25 °C × 0.95 aw treatments + F. graminearum inoculation. This was reflected in the total accumulated CO2 in the treatments. The maximum DMLs were in the 0.95 aw/20–25 °C treatments and at 10 °C/0.95 aw. The DMLs were modelled to produce contour maps of the environmental conditions resulting in maximum/minimum losses. Contamination with ZEN/ZEN-related compounds were quantified. Maximum production was at 25 °C/0.95–0.93 aw and 20 °C/0.95 aw. ZEN contamination levels plotted against DMLs for all the treatments showed that at ca. 1.0% DML, the risk was high. This type of data is important in building a database for the development of a post-harvest decision support system for relative risks of different mycotoxins.

Changes in environmental stress impact on secondary metabolite (SM) production profiles. Few studies have examined targeted SM production patterns in relation to interacting environmental conditions in stored cereals. The objectives were to examine the effect of water activity (aw; 0.95–0.90) x temperature (10–25 °C) on SM production on naturally contaminated stored wheat and that inoculated with Fusarium graminearum. Samples were analysed using Liquid Chromatography-Tandem Mass Spectrometry (LC-MS/MS) on (a) total number of known SMs, (b) their concentrations and (c) changes under environmental stress. 24 Fusarium metabolites were quantified. Interestingly, statistical differences (ChisSq., p < 0.001) were observed in the number of SMs produced under different sets of interacting environmental conditions. The dominant metabolites in natural stored grain were deoxynivalenol (DON) and nivalenol (NIV) followed by a range of enniatins (A, A1, B, B1), apicidin and DON-3-glucoside at 10 °C. Increasing temperature promoted the biosynthesis of other SMs such as aurofusarin, moniliformin, zearalenone (ZEN) and their derivatives. Natural wheat + F. graminearum inoculation resulted in a significant increase in the number of metabolites produced (ChisSq., p < 0.001). For ZEN and its derivatives, more was produced under cooler storage conditions. Fusarin C was enhanced in contrast to that for the enniatin group. The relative ratios of certain groups of targeted SM changed with environmental stress. Both temperature and aw affected the amounts of metabolites present, especially of DON and ZEN. This study suggests that the dominant SMs produced in stored temperate cereals are the mycotoxins for which legislation exists. However, there are changes in the ratios of key metabolites which could influence the relative contamination with individual compounds. Thus, in the future, under more extreme environmental stresses, different dominant SMs may be formed which could make present legislation out of step with the future ...

Aflatoxins (AFs) are produced by fungi in crops and can cause liver cancer. Permitted levels are legislated and batches of grain are rejected based on average concentrations. Corn grown in Southern Georgia (GA), USA, which experiences drought during the mid-silk growth period in June, is particularly susceptible to infection by Aspergillus section Flavi species which produce AFs. Previous studies showed strong association between AFs and June weather. Risk factors were developed: June maximum temperatures > 33 °C and June rainfall 33 °C and rainfall < 50 mm increased and then plateaued for both emissions scenarios. The percentage of years thresholds were exceeded was greater for RCP 8.5 than RCP 4.5. The spatial distribution of high-risk counties changed over time. Results suggest corn growth distribution should be changed or adaptation strategies employed like planting resistant varieties, irrigating and planting earlier. There were significantly more counties exceeding thresholds in 2010–2040 compared to 2000–2030 suggesting that adaptation strategies should be employed as soon as possible.

Aflatoxins (AFs) are produced by fungi in crops and can cause liver cancer. Permitted levels are legislated and batches of grain are rejected based on average concentrations. Corn grown in Southern Georgia (GA), USA, which experiences drought during the mid-silk growth period in June, is particularly susceptible to infection by Aspergillus section Flavi species which produce AFs. Previous studies showed strong association between AFs and June weather. Risk factors were developed: June maximum temperatures > 33 °C and June rainfall 33 °C and rainfall < 50 mm increased and then plateaued for both emissions scenarios. The percentage of years thresholds were exceeded was greater for RCP 8.5 than RCP 4.5. The spatial distribution of high-risk counties changed over time. Results suggest corn growth distribution should be changed or adaptation strategies employed like planting resistant varieties, irrigating and planting earlier. There were significantly more counties exceeding thresholds in 2010–2040 compared to 2000–2030 suggesting that adaptation strategies should be employed as soon as possible.

Changes in environmental stress impact on secondary metabolite (SM) production profiles. Few studies have examined targeted SM production patterns in relation to interacting environmental conditions in stored cereals. The objectives were to examine the effect of water activity (aw; 0.95–0.90) x temperature (10–25 °C) on SM production on naturally contaminated stored wheat and that inoculated with Fusarium graminearum. Samples were analysed using Liquid Chromatography-Tandem Mass Spectrometry (LC-MS/MS) on (a) total number of known SMs, (b) their concentrations and (c) changes under environmental stress. 24 Fusarium metabolites were quantified. Interestingly, statistical differences (ChisSq., p < 0.001) were observed in the number of SMs produced under different sets of interacting environmental conditions. The dominant metabolites in natural stored grain were deoxynivalenol (DON) and nivalenol (NIV) followed by a range of enniatins (A, A1, B, B1), apicidin and DON-3-glucoside at 10 °C. Increasing temperature promoted the biosynthesis of other SMs such as aurofusarin, moniliformin, zearalenone (ZEN) and their derivatives. Natural wheat + F. graminearum inoculation resulted in a significant increase in the number of metabolites produced (ChisSq., p < 0.001). For ZEN and its derivatives, more was produced under cooler storage conditions. Fusarin C was enhanced in contrast to that for the enniatin group. The relative ratios of certain groups of targeted SM changed with environmental stress. Both temperature and aw affected the amounts of metabolites present, especially of DON and ZEN. This study suggests that the dominant SMs produced in stored temperate cereals are the mycotoxins for which legislation exists. However, there are changes in the ratios of key metabolites which could influence the relative contamination with individual compounds. Thus, in the future, under more extreme environmental stresses, different dominant SMs may be formed which could make present legislation out of step with the future contamination which might occur.

Poor storage of cereals, such as maize can lead to both nutritional losses and mycotoxin contamination. The aim of this study was to examine the respiration of maize either naturally contaminated or inoculated with Aspergillus flavus to examine whether this might be an early and sensitive indicator of aflatoxin (AF) contamination and relative storability risk. We thus examined the relationship between different interacting storage environmental conditions (0.80–0.99 water activity (aw) and 15–35°C) in naturally contaminated and irradiated maize grain + A. flavus on relative respiration rates (R), dry matter losses (DMLs) and aflatoxin B1 and B2 (AFB1-B2) contamination. Temporal respiration and total CO2 production were analysed by GC-TCD, and results used to calculate the DMLs due to colonisation. AFs contamination was quantified at the end of the storage period by HPLC MS/MS. The highest respiration rates occurred at 0.95 aw and 30–35°C representing between 0.5% and 18% DMLs. Optimum AFs contamination was at the same aw at 30°C. Highest AFs contamination occurred in maize colonised only by A. flavus. A significant positive correlation between % DMLs and AFB1 contamination was obtained (r = 0.866, p < 0.001) in the irradiated maize treatments inoculated with A. flavus. In naturally contaminated maize + A. flavus inoculum loss of only 0.56% DML resulted in AFB1 contamination levels exceeding the EU legislative limits for food. This suggests that there is a very low threshold tolerance during storage of maize to minimise AFB1 contamination. This data can be used to develop models that can be effectively used in enhancing management for storage of maize to minimise risks of mycotoxin contamination.