Assessment of the Effect of Satureja montana and Origanum virens Essential Oils on Aspergillus flavus Growth and Aflatoxin Production at Different Water Activities

© 2020 by the authors. Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (http://creativecommons.org/licenses/by/4.0/).Aflatoxin contamination of foodstuffs poses a serious risk to food security, and it is essential to search for new control methods to prevent these toxins entering the food chain. Several essential oils are able to reduce the growth and mycotoxin biosynthesis of toxigenic species, although their efficiency is strongly influenced by the environmental conditions. In this work, the effectiveness of Satureja montana and Origanum virens essential oils to control Aspergillus flavus growth was evaluated under three water activity levels (0.94, 0.96 and 0.98 aw) using a Bioscreen C, a rapid in vitro spectrophotometric technique. The aflatoxin concentrations at all conditions tested were determined by HPLC-FLD. Aspergillus flavus growth was delayed by both essential oil treatments. However, only S. montana essential oil was able to significantly affect aflatoxin production, although the inhibition percentages widely differed among water activities. The most significant reduction was observed at 0.96 aw, which is coincident with the conditions in which A. flavus reached the highest levels of aflatoxin production. On the contrary, the treatment with S. montana essential oil was not effective in significantly reducing aflatoxin production at 0.94 aw. Therefore, it is important to study the interaction of the new control compounds with environmental factors before their application in food matrices, and in vitro ecophysiological studies are a good option since they provide accurate and rapid results.Peer reviewedFinal Published versio

Interacting Abiotic Factors Affect Growth and Aflatoxin B1 Production Profiles of Aspergillus flavus Strains on Pistachio-Based Matrices and Pistachio Nuts

© 2021 Baazeem, Garcia-Cela, Medina and Magan. This is an openaccess article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.Pistachio nuts are an economically important commodity produced by many countries. They can be colonized by mycotoxigenic fungi, especially Aspergillus flavus, resulting in contamination with aflatoxins (AFs), especially aflatoxin B1 (AFB1), a Class 1a carcinogen. The objectives were to examine the effect of interactions between the two key abiotic factors, temperature and water activity (aw) on (a) in vitro growth and AFB1 production by four strains of A. flavus isolated from pistachio nuts, on a milled pistachio nut medium modified ionically (NaCl) and non-ionically (glycerol) in the range 20–35°C and 0.995–0.85 aw, (b) colonization of layers of raw pistachio nuts stored at different interacting temperature x aw conditions and on relative AFB1 production and (c) develop models to produce contour maps of the optimal and marginal boundary conditions for growth and AFB1 production by up to 4 strains of this species. On pistachio nut-based media, optimum growth of four strains of A. flavus was at 0.98–0.95 aw and 30–35°C. Optimum AFB1 production was at 30–35°C and 0.98 aw. No significant differences in growth was found on ionic and non-ionically modified media. Colonization of layers of raw pistachio nuts was slower and contamination with AFB1 significantly less than in in vitro studies. Contour maps based on the pooled data for up to four strains (in vitro, in situ) showed the optimum and marginal conditions for growth and AFB1 production. These data can be used to identify those conditions which represent a high, intermediate or low risk of colonization and AFB1 contamination in the pistachio nut processing chain. These results are discussed in the context of the development of appropriate intervention strategies to minimize AFB1 contamination of this economically important commodity.Peer reviewedFinal Published versio

Application of Risk Management Metrics for ochratoxin-A control in the coffee chain

© 2023 The Authors. Published by Elsevier Ltd. This is an open access article under the CC BY license (http://creativecommons.org/licenses/by/4.0/).Coffee is one of the most consumed beverages worldwide. Like other agricultural products, coffee is susceptible of colonization by mycotoxin-producing fungi and therefore, the presence of mycotoxins. These chemical hazards can pose a risk for consumers, as some of them are potentially carcinogenic, neurotoxic, or immunosuppressive. Several countries worldwide have established maximum legal limits on the final product regarding ochratoxin A (OTA), but this leads to inefficiencies in the coffee value chain, as there is uncertainty if a batch may be under the legal limits at the time of reaching the consumers. The application of Risk Management Metrics can be a useful tool for managers to forecast if a particular batch of coffee could be suitable for a determined target market. In this study, a cross-case analysis of the coffee production chain was performed, and quantitative thresholds were established along the different steps. This information can provide managers with up-to-date information regarding the potential use of each batch, minimizing food waste, assuring food safety, and improving chain efficiency.Peer reviewe

Electrospinning Alginate/Polyethylene Oxide and Curcumin Composite Nanofibers

© 2020 Elsevier Ltd. All rights reserved. This manuscript is licensed under the Creative Commons Attribution-NonCommercial-NoDerivatives 4.0 International Licence http://creativecommons.org/licenses/by-nc-nd/4.0/.Manufacturing a sodium alginate (SA) and polyethylene oxide (PEO) composite loaded with curcumin (CU) was accomplished in this study by using electrospinning. These composite nanofibers were crosslinked using trifluoroacetic acid (TFA) mechanically characterized along with the morphological properties of the composite nanofiber mesh. We were successful in manufacturing the composite nanofibers with a wide range of CU concentrations ranging from 10 to 40 wt%. Firstly, dissolved in a saturated water/CU solution it was added to SA/PEO blending, homogenized and electrospun. Mechanical properties were affected by both CU addition and the cross-linking process, resulting in a higher ultimate tensile stress (MPa) (from 4.3±2 to 15.1±2 at 10% CU) and Young modulus (GPa) (0.0076±0.003, 0.044±0.003 before and after TFA). CU was successfully encapsulated in the SA nanofibers and excellent mechanical properties were obtained. By using a biocompatible TFA crosslinking and the natural properties of alginate this nanofiber composite could potentially be used for filtering, environmental pollution control, food packaging and for tissue engineering.Peer reviewe

Influence of two garlic-derived compounds, propyl propane thiosulfonate (PTS) and propyl propane thiosulfinate (PTSO), on growth and mycotoxin production by Fusarium species in vitro and in stored cereals

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

Influence of two garlic-derived compounds, propyl propane thiosulfonate (PTS) and propyl propane thiosulfinate (PTSO), on growth and mycotoxin production by Fusarium species in vitro and in stored cereals

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

Fusarium graminearum in stored wheat: use of CO2 production to quantify dry matter losses and relate this to relative risks of Zearalenone contamination under interacting environmental conditions

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

Biological control products for aflatoxin prevention in Italy: Commercial field evaluation of atoxigenic Aspergillus flavus active ingredients

Since 2003, non-compliant aflatoxin concentrations have been detected in maize produced in Italy. The most successful worldwide experiments in aflatoxin prevention resulted from distribution of atoxigenic strains of Aspergillus flavus to displace aflatoxin-producers during crop development. The displacement results in lower aflatoxin concentrations in harvested grain. The current study evaluated in field performances of two atoxigenic strains of A. flavus endemic to Italy in artificially inoculated maize ears and in naturally contaminated maize. Co-inoculation of atoxigenic strains with aflatoxin producers resulted in highly significant reductions in aflatoxin concentrations (>90%) in both years only with atoxigenic strain A2085. The average percent reduction in aflatoxin B1 concentration in naturally contaminated maize fields was 92.3%, without significant differences in fumonisins between treated and control maize. The vegetative compatibility group of A2085 was the most frequently recovered A. flavus in both treated and control plots (average 61.9% and 53.5% of the A. flavus, respectively). A2085 was therefore selected as an active ingredient for biocontrol products and deposited under provisions of the Budapest Treaty in the Belgian Co-Ordinated Collections of Micro-Organisms (BCCM/MUCL) collection (accession MUCL54911). Further work on development of A2085 as a tool for preventing aflatoxin contamination in maize produced in Italy is ongoing with the commercial product named AF-X1™

Solute and matric potential stress on Penicillium verrucosum : Impact on growth, gene expression and ochratoxin A p production

Funding Information: S.A. is grateful to the British Council and the Newton Musharraf Programme for financial support. Publisher Copyright: © 2020 Wageningen Academic Publishers. All rights reserved. Copyright: Copyright 2020 Elsevier B.V., All rights reserved.Penicillium verrucosum survives in soil and on cereal debris. It colonises grain during harvesting, drying and storage. There is no information on the relative tolerance of P. verrucosum to solute and matric stress in terms of colonisation, or on the biosynthetic toxin gene clusters or ochratoxin A (OTA) production. The objectives were to examine the effect of ionic and non-ionic solute and matric potential stress on (a) growth, (b) expression of two toxin biosynthetic genes otapksPV and otanrpsPV, and (c) OTA production by a strain of P. verrucosum. Optimum growth and OTA production were at-7.0 MPa (= 0.95 water activity, aw) and-1.4 MPa (= 0.99 aw), respectively, regardless of whether solute (Ψs) or matric (Ψm) stress was imposed. P. verrucosum was more sensitive to ionic solute stress (NaCl) with no growth at-19.6 MPa (=0.86 aw) while growth still occurred in the non-ionic solute (glycerol) and matric stress treatments. Relative gene expression of the biosynthetic genes using PCR (RT-qPCR) showed that the otapksPV gene was expressed over a wide range of ionic/non-ionic solute stress conditions (-1.4 to-14.0 MPa; = 0.99-0.90 aw). The highest expression was in the non-ionic Ψs stress treatments at-7.0 MPa (= 0.95 aw). However, the otanrpsPV gene was significantly up regulated under Ψm stress, especially with freely available water (-1.4 MPa = 0.99 aw). OTA production was significantly decreased as Ψs or Ψm stress were imposed. Limited OTA production occurred in the driest treatments under Ψs and Ψm stress respectively. The impact of these two types of stresses on the growth of P. verrucosum was quite different from that for OTA production. The results are discussed in the context of the life cycle and ecological characteristics of this species in contaminating cereals with OTA in the post-harvest phase of the cereal chain.Peer reviewedFinal Accepted Versio

Fungal diversity and metabolomic profiles in GM and isogenic non-GM maize cultivars from Brazil

The Author(s) 2020. This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons licence, and indicate if changes were made. To view a copy of this licence, http://creativecommons.org/licenses/by/4.0/.There is little knowledge of the microbial diversity, mycotoxins and associated secondary metabolites in GM maize and isogenic non-GM cultivars (cvs). This study has quantified the microbial populations and dominant fungal genera in 6 cvs of each type representative of herbicide, pesticide or stacked resistance to both. The predominant mycotoxins and targeted metabolomics profiles were also compared between the two sets of cvs. This showed that the overall fungal populations were 8.8 CFUs g−1 maize. The dominant genera, isolated from maize samples, whether surface-sterilised or not, in all maize cvs were Fusarium, followed by Penicillium, Aspergillus and occasionally Cladosporium and Alternaria. The analysis of the targeted metabolomics showed that approx. 29 different metabolites were detected. These were dominated by fumonisins and minor Penicillium spp. metabolites (questiomycin A and rugulovasine A). Interestingly, the range and number of mycotoxins present in the GM cvs were significantly lower than in the non-GM maize samples. This suggests that while the fungal diversity of the two types of maize appeared to be very similar, the major contaminant mycotoxins and range of toxic secondary metabolites were much lower in the GM cvs.Peer reviewedFinal Published versio