Browsing by Author "Magan, Naresh"
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Item Open Access Acclimatisation of Fusarium langsethiae, F. poae and F. sporotrichioides to elevated CO2: impact on fungal growth and mycotoxin production on oat-based media(Elsevier, 2023-03-28) Kahla, Amal; Verheecke-Vaessen, Carol; Delpino-Deelias, Mariluz; Gutierrez-Pozo, Maria; Medina, Angel; Magan, Naresh; Doohan, FionaOats are highly susceptible to infection by Fusarium species, especially F. langsethiae, F. poae and F. sporotrichioides which contaminate the grain with mycotoxins. Climate change is expected to affect fungal colonisation and associated mycotoxin production. The objective of this study was to examine the effect of acclimatisation to elevated CO2 on the growth and mycotoxin production capacity of these fungal species. Strains of F. langsethiae (FL; seven strains), F. poae (FP; two strains) and F. sporotrichioides (FS; one strain) were acclimatised by sub-culturing for 10 generations at either 400 or 1000 ppm CO2 under diurnal temperature conditions. At each sub-culturing, the effect of acclimatisation to elevated CO2 on (a) lag phase prior to growth, (b) growth rate on oat-based media was assessed. Additionally, the production of type A trichothecenes and related toxic secondary metabolites of sub-cultures after 1, 7 and 10 generations were assessed using LC-MS/MS qTRAP. The results showed that Fusarium strains had an increased lag time and growth rate in response to the combined effect of sub-culturing and elevated CO2 levels. T-2 + HT-2 production was affected by elevated CO2 in strain FL4 (7.1-fold increase) and a decrease in strain FL1 (2.0-fold decrease) at the first sub-culturing and FS (1.3-fold decrease) after 7 sub-cultures compared to ambient conditions. The effect of sub-culturing on T-2 + HT-2 production varied depending on the fungal strain. For strain FL4, significantly less T-2 + HT-2 toxins were produced after 10 generations (4.4-fold decrease) as compared to that under elevated CO2 conditions after one sub-culture, and no change was observed under ambient conditions. The FS strain showed significant stimulation of T-2 + HT-2 toxin production after 10 sub-cultured generations (1.1-fold increase) compared to the initial sub-culture of this strain under elevated CO2 conditions. The production of other toxic secondary metabolites was generally not impacted by elevated CO2 conditions or by sub-culture for 10 generations, with the exceptions of FL1 and FP1. FL1 produced significantly more neosolaniol after 10 generations, when compared to those after 1 and 7, regardless of the CO2 conditions. For FP1, elevated CO2 significantly triggered beauvericin production after an initial sub-culture when compared to ambient conditions at the same sub-culture stage (29-fold). FP1 acclimatisation to elevated CO2 led to a decrease of beauvericin production after 10 generations when compared to 1 (6-fold). In contrast, sub-culturing for 10 generations compared to 1 under ambient CO2 conditions resulted in an increase in this toxin (12-fold).Item Open Access Advances in molecular and genomic research to safeguard food and feed supply from aflatoxin contamination(Wageningen Academic Publishers, 2018-02-23) Bhatnagar, D; Rajasekaran, K; Gilbert, M; Cary, J. W.; Magan, NareshWorldwide recognition that aflatoxin contamination of agricultural commodities by the fungus Aspergillus flavus is a global problem has significantly benefitted from global collaboration for understanding the contaminating fungus, as well as for developing and implementing solutions against the contamination. The effort to address this serious food and feed safety issue has led to a detailed understanding of the taxonomy, ecology, physiology, genomics and evolution of A. flavus, as well as strategies to reduce or control pre-harvest aflatoxin contamination, including (1) biological control, using atoxigenic aspergilli, (2) proteomic and genomic analyses for identifying resistance factors in maize as potential breeding markers to enable development of resistant maize lines, and (3) enhancing host-resistance by bioengineering of susceptible crops, such as cotton, maize, peanut and tree nuts. A post-harvest measure to prevent the occurrence of aflatoxin contamination in storage is also an important component for reducing exposure of populations worldwide to aflatoxins in food and feed supplies. The effect of environmental changes on aflatoxin contamination levels has recently become an important aspect for study to anticipate future contamination levels. The ability of A. flavus to produce dozens of secondary metabolites, in addition to aflatoxins, has created a new avenue of research for understanding the role these metabolites play in the survival and biodiversity of this fungus. The understanding of A. flavus, the aflatoxin contamination problem, and control measures to prevent the contamination has become a unique example for an integrated approach to safeguard global food and feed safety.Item Open Access Alternaria in food: Ecophysiology, mycotoxin production and toxicology- Erratum(Hangug Gynnhaghoi, 2015-09-01) Lee, Hyang Burm; Patriarca, Andrea; Magan, NareshItem Open Access Applications of volatile fingerprint sensor arrays for rapid detection of environmental contaminants(Cranfield University, 2005-03) Canhoto, Olinda; Aldred, David; Magan, NareshThe electronic nose (e-nose) technology has rapidly evolved in the past decade with a range of applications in the food industry, medical diagnosis, and recently environmental monitoring. This is the first time that this technology has been examined in detail for a range of specific environmental applications including: detection of low concentrations of bacterial, fungal and heavy metal contaminants in potable water; analyses of changes in the microbial activity of soil samples amended with heavy metals; and the detection of fungal contaminants in paper samples from library material. In some studies comparisons between different e-nose systems has also been carried out. The e-nose system based on a conducting polymer (CP) sensor array Bloodhound (BH114) was able to detect different bacterial species (Escherichia coli, Pseudomonas aeruginosa and Enterobacter aerogenes), initially inoculated in tap, reverse osmosis and bottled water with a concentration of 102 cells mL-1, after 24 hrs incubation. In the presence of low concentrations (0.5 ppm) of a mixture of heavy metal ions including cadmium, lead and zinc, the volatile pattern produced by the bacterial species was discriminated from that where no metal was added, probably due to a change in the microbial metabolism. The Bloodhound e-nose system was also used to detect fungal spores of Aspergillus fumigatus, Fusarium culmorum and a Penicillium species, inoculated in water samples. The initial concentrations were 102 – 105 spores mL-1. Good discrimination was observed between the control samples after 24 hrs incubation at 25oC. After 48 hrs incubation, it was possible to differentiate between the various spore concentrations present in water samples. Good reproducibility was achieved as results from different days were consistent and data could be pooled and combined for analysis. A comparative study was performed with three e-nose instruments, two of them had CP sensor arrays (Bloodhound (BH-114); Neotronics (eNOSE 4000), and the third was a metal oxide (MO) sensor-based system, the NST 3220. The experiments carried out with the CP based-systems showed similar results when analysing water samples contaminated with 104 and 102 bacterial cells mL-1 after 24 hrs incubation. Both CP and MO based e-nose systems could differentiate control water samples from those contaminated with both bacteria and fungal spores. GC-SPME analyses confirmed the results obtained with the e-nose system of metal ions and bacterial cells in water samples. At-line studies were performed with the MO array-based system (NST 3220), for the detection of contamination episodes. E. coli and P. aeruginosa cells were used as contamination agents for tap and reverse osmosis sterile water, in two concentration levels, 102 and 106 cells mL-1. The samples collected downstream in a simulated watercourse, were analysed by the e-nose over a period of 1-2 hrs. The results suggested the potential of this technique to monitor episodes of bacterial cells at a low concentration in water samples. Experiments performed in soil samples artificially and naturally contaminated with heavy metal ions were analysed with the MO-based e-nose system. Results indicated that for artificially contaminated soil samples, after 40 days incubation the control samples could be discriminated from those containing 3 and 100 ppm of metal ions. For naturally contaminated soils, the sensor array was only able to separate samples containing a high concentrations of metal ions. Headspace analysis of cellulose-based agar showed good discrimination between Aspergillus terreus, A. hollandicus and Eurotium chevallieri, after 20 hrs incubation at 25oC. An increase in the incubation period to 40 hrs resulted in better separation between the control and fungal treatments. In situ studies performed on paper samples suggested that the e-nose was able to discriminate between control samples and paper inoculated with 103 fungal spores mL-1. The substrate was a determinant factor in the headspace analysis of microbial species. It was shown that the same fungal species produced different volatile profiles according to the growth substrate.Item Open Access Aspergillus species and mycotoxins: occurrence and importance in major food commodities(Elsevier, 2018-05-18) Taniwaki, Marta Hiromi; Pitt, John I; Magan, NareshAspergillus species produce important mycotoxins, in particular aflatoxins, produced by A. flavus and related species, and ochratoxin A, produced by A. ochraceus and related species and also A. carbonarius and (less commonly) A. niger. In this review we briefly discuss the distribution of toxigenic Aspergillus species in nuts, coffee and cocoa beans, dried fruits, grapes, maize, rice and small grains. Future perspectives of distribution of Aspergillus species in foods are briefly discussed taking into account the impacts of climate change and the resilience of these mycotoxigenic species.Item Open Access Assessment of intraspecies variability in fungal growth initiation of Aspergillus flavus and aflatoxin B1 production under static and changing temperature levels using different initial conidial inoculum levels(Elsevier, 2018-02-16) Aldars-García, Laila; Marín, Sonia; Sanchis, Vicente; Magan, Naresh; Medina-Vayá, ÁngelIntraspecies variability in fungal growth and mycotoxin production has important implications for food safety. Using the Bioscreen C we have examined spectrophotometrically intraspecies variability of A. flavus using 10 isolates under different environments, including temperature shifts, in terms of growth and aflatoxin B1 (AFB1) production. Five high and five low AFB1 producers were examined. The study was conducted at 5 isothermal conditions (from 15 to 37 °C) and 4 dynamic scenarios (between 15 and 30 °C). The experiments were carried out in a semisolid YES medium at 0.92 aw and two inoculum levels, 102 and 103 spores/mL. The Time to Detection (TTD) of growth initiation was determined and modelled as a function of temperature through a polynomial equation and the model was used to predict TTD under temperature upshifts conditions using a novel approach. The results obtained in this study have shown that a model can be developed to describe the effect of temperature upshifts on the TTD for all the studied isolates and inoculum levels. Isolate variability increased as the growth conditions became more stressful and with a lower inoculum level. Inoculum level affected the intraspecies variability but not the repeatability of the experiments. In dynamic conditions, isolate responses depended both on the temperature shift and, predominantly, the final temperature level. AFB1 production was highly variable among the isolates and greatly depended on temperature (optimum temperature at 30–35 °C) and inoculum levels, with often higher production with lower inoculum. This suggests that, from an ecological point of view, the potential isolate variability and interaction with dynamic conditions should be taken into account in developing strategies to control growth and predicting mycotoxin risks by mycotoxigenic fungi.Item Open Access Assessment of occupational exposure to gasoline vapour at petrol stations(Cranfield University, 2016-01) Alyami, Ahmed Rashid; Crump, Derrick; Walton, Christopher; Magan, Naresh; Salama, KhaledPetrol station attendants’ exposure to gasoline vapours while refuelling vehicles has raised health concerns, especially in tropical countries like Saudi Arabia. This is due to the increase of gasoline vaporisation by the high temperatures and related weather conditions. This represents an increase risk of inhaling more vapours than its counterpart temperate countries. Furthermore, exposure during extended working hours (12 hrs shifts), with no vapour recovery system and the handling of gasoline containing a high percentage of volumes of toxic substances (e.g. BTEXs) have not been adequately addressed previously in Saudi Arabia. Therefore, this study was designed and carried out to investigate the validity of this concern by assessing and quantifying full shift exposures to gasoline vapours during the petrol filling process. Different exposure assessment methodologies were employed and evaluated for their suitability. The study assessed the exposures of 41 attendants via passive, active, and direct reading methods at twelve petrol stations with both high and low sales in the Eastern Province of Saudi Arabia. The study was conducted during the winter and summer months to test the seasonal variation of the pattern of exposure. The effects of the quantity of gasoline sold, the locations of the stations, weather variations (e.g. wind speed, temperature, and humidity) were tested. A purpose built mini-weather stations and modified thermometres were utilized to accurately monitor the prevailing weather conditions. Forward-looking infrared (FLIR) thermal image cameras were utilised to visualise the size and movement behaviour of the vapour plumes during petrol refuelling. Furthermore, analytical lab trials were carried out to characterise the gasoline vapour component under different temperatures. These were used to propose a new OEL. The geometric means of the personal passive results for BTEX and MTBE (0.18 ppm, 0.24 ppm, 0.09 ppm, 0.18 ppm, 1.57 ppm, respectively) were found to be relatively higher than those reported previously for Europe and North America. These results are discussed in the context of the impact that such exposure will have on people involved in this industry in petrol stations in Saudi Arabia.Item Open Access Bacteria in heavy metal contaminated soil: diversity, tolerance and use in remediation(Cranfield University, 2010) Pires, Carlos; Magan, Naresh; Castro, PaulaThe objectives of this project were to determine the bacterial diversity in a heavily contaminated metal region of Portugal. Both traditional and molecular based methods were used to identify tolerant strains and species. The most tolerant species were subsequently identified and utilized for examining the potential for using them to immobilize specific metals from contaminated waste streams by comparing different support materials. Heterotrophic bacterial populations were isolated and characterized from a contaminated industrial area in Northern Portugal. In a first sampling, 278 strains were isolated in different solid media. To assess the diversity of this ecological site and to select representative strains, the isolates were screened by using Random Amplified Polymorphic DNA (RAPD)-PCR profiles. Phenotypic characterization, phylogenetic analysis by sequencing the 16S rRNA genes and metal tolerance tests with zinc (Zn), cadmium (Cd) and arsenic (As) were performed with the selected strains. Recovered gram-positive isolates were related to class Actinobacteria and Bacilli. The majority of the isolates were related to genera Microbacterium and Bacillus. Strains from the genus Arthrobacter were also well represented. 16S rRNA gene sequence similarity of the gram-negative isolates showed that they were related to classes γ-Proteobacteria, ß-Proteobacteria, a-Proteobacteria and Flavobacteria. The most frequently isolated taxa were γ-Proteobacteria, related with the genus Pseudomonas, where a large number of isolates were clustered. These genera are common in metal contaminated environments. Many of the strains (approx. 17) had a high level of tolerance to the heavy metals tested. A total of 13 isolates were not able to grow when metals were present. In a second sampling the soil rhizosphere was screened for bacterial populations, using metal-based selective media for isolation. About 42 strains were recovered when metal supplemented media was used. The gram-positive population were predominantly Bacilli and Actinobacteria members. Bacillus, Microbacterium and Arthrobacter were the most common gram-positive genera. Gram-negative genera were from the same classes as in the first sampling however Sphingobacteria was present. γ-Proteobacteria and ß-Proteobacteria were the most common taxa. The isolates were shown to be very resistant to Zn and As, with about half of the isolates able to grow with Cd present. Interestingly, no strains could grow in the presence of metal mixtures. Despite the number of strains recovered in both samplings the majority of the isolates were clustered within a very small number of genera. During the sampling periods two strains showing low similarity to other bacteria were isolated. These strains were characterized and studied in detail justifying their classification as representing two novel species of the genus Chryseobacterium. The names proposed for these organisms are Chryseobacterium palustre sp. nov. (type strain 3A10[type strain) ) and Chryseobacterium humi sp. nov. (type strain ECP37[type strain] ). Three isolates 1C2, 1ZP4 and EC30 belonging to genera Cupriavidus, Sphingobacterium and Alcaligenes respectively, showing high tolerance to heavy metals, were selected for further study in immobilised systems for Zn and Cd removal. In most cases, matrices (alginate, pectate and a synthetic cross-linked polymer) with immobilised bacteria showed better metal removal. 1C2, a strain belonging to the Cupriavidus genera, was able to increased the removal of Zn; EC30, a bacteria related to Alcaligenes, was the most promising candidate for Cd removal, especially when combined with the synthetic polymer. Removal of metals as single or in binary mixtures was also assessed. Cd removal was most effective when single metal solutions were tested using immobilised bacteria and examining metal matrixes. Based on the strains used and the matrices tested, best results were obtained for removal of Zn from binary mixtures with Cd. Potential exists for further studies to exploit these bacterial strain to develop effective bioremediation approaches for the removal of heavy metals from waste water streams.Item Open Access Biocontrol of mycotoxins: dynamics and mechanisms of action(Elsevier, 2017-09-18) Medina-Vayá, Ángel; Mohale, Sejakhosi; Samsudin, Nik Iskandar Putra; Rodriguez-Sixtos, Alicia; Rodriguez, Alicia; Magan, NareshThis paper discusses the relationship between biocontrol agents (BCAs) and mycotoxigenic fungi and mycotoxin control. In most cases BCAs are examined for control of growth of fungal pathogens and disease symptoms. However, for mycotoxin control the approach and focus needs to be different. The mechanism of action and the inoculum dose necessary for control of toxin production by Aspegillus, Penicillium and Fusarium species may be different from that for traditional fungal plant pathogens. The mechanisms of action, the relative inoculum potential and the impact that interacting environmental conditions have on control of key components of the life cycle of mycotoxigenic fungi are considered. The practical aspects of production and formulation hurdles are discussed and potential future approaches and strategies which may need to be considered for more effective biocontrol of mycotoxigenic fungi and mycotoxins are presented.Item Open Access Biodiversity of mycotoxigenic aspergillus species in Egyptian peanuts and strategies for minimizing aflatoxin contamination(Cranfield University, 2010-12) Sultan, Yousef Yasseen Abdel-Rahmin; Magan, NareshPeanuts are an important crop grown in Egypt for either local consumption or export to European markets. The present study examined the importance of mycotoxigenic Aspergilli in Egyptian peanuts from five different regions (Alexandria, El-Beheira, El- Daqahliya, El-Sharqiya, Asyut) in two seasons (2007, 2008). This led to consideration of different potential strategies to control aflatoxigenic A. flavus strains and associated aflatoxin contamination of peanuts. The most common species in peanuts were from Aspergillus section Flavi, Aspergillus section Nigri and Aspergillus section Circumdati. Both qualitative (coconut cream agar) and quantitative analyses (HPLC) were used to analyse the potential mycotoxin production by strains isolated from peanuts. Of a total of 88 Aspergillus section Flavi strains examined, 90% were aflatoxigenic. Cont/d.Item Open Access Bioremediation of the pesticides Dieldrin, Simazine, Trifluralin using tropical and temperate white-rot fungi(Cranfield University, 1997-08) Elyassi, Ali; Magan, NareshThe natural breakdown of three pesticides on the UK Red Lis~ dieldrin~ simazine and trifluralin in water and soil varied with environmental conditions. In both sterile and unsterile water trifluralin was degraded to some extent at 20 and 30°C. In contras~ dieldrin and simazine were stable over the 42 days incubation period. A gradient HPLC method was developed for the simultaneous quantification of the three pesticides in soil. In field capacity soil mixtures of the three pesticides (5 and 10 ppm) showed a similar stability with limited degradation at 20°C but increased rates of degradation at 30°C. At the higher concentration the pesticides naturally degraded at a slower rate. Simazine and trifluralin degradation was significantly enhanced with increasing temperature from 20 to 30°C. Water potential (field capacity~ -0.065 MPa~ and - 0.28 MPa) had little effect on the natural breakdown rate of dieldrin. Simazine showed a greater breakdown in the mid-wetness soil~ while trifluralin was degraded rapidly in the field capacity soil, but not at all in the driest treatment over the 70 day experimental period. In vitro studies on solid agar media overlayed with cellophane showed that of four fungi examined~ Trametes cingulata, Trametes socotrana (tropical species) and Phanerochaete chrysosporium and Polystictus versicolor (temperate species) all except P.chrysosporium were able to grow in the presence of 5 ppm of any of the three pesticides at 20 and 30°C, with the latter only growing at 30°C. At 10 ppm concentration P. chrysosporium did not grow, regardless of temperature or time of incubation (up to 56 days). HPLC was used to quantify the temporal rates of degradation in the solid agar media and this showed that P. versicolor and T. socotrana were very effective at breaking down the three pesticides, at 20 and 30°C. The chosen fungi were grown on chopped straw as a carrier and incorporated into soil microcosms in the ratio of 1:10 containing mixtures of the three pesticides (5, 10 ppm) at 20 and 30°C, and subsequently under different water potential regimes at 20°C only, over periods of 70 days. P. versicolor alone significantly increased breakdown of 5 ppm dieldrin by 26% over untreated controls, while simazine breakdown was increased by 16%. However, for simazine at 30°C there was no difference between temporal rates of natural breakdown and those containing fungal inocula, regardless of concentration. With 5 ppm trifluralin, a maximum breakdown in untreated soil was 67% after 70 days. By contras~ this pesticide was undetectable after 28 days in the presence of the inoculant P . versicolor. This increased to 42 days where a mixture of the two fungi were used. Generally the mixture of fungi used in this study were not as effective in bioremediation of these pesticides as a single species. Field capacity soil appeared to be the best condition for P. versicolor to degrade dieldrin and trifluralin added at 10 ppm. However, for simazine this occurred in the driest water potential (-0.28 MPa) used.Item Open Access Carbon dioxide mediates the response to temperature and water activity levels in Aspergillus flavus during infection of maize kernels(MDPI, 2017-12-22) Gilbert, Matthew K.; Medina-Vayá, Ángel; Mack, Brian M.; Lebar, Matthew D.; Rodriguez, Alicia; Bhatnagar, Deepak; Magan, Naresh; Obrian, Gregory; Payne, GaryAspergillus flavus is a saprophytic fungus that may colonize several important crops, including cotton, maize, peanuts and tree nuts. Concomitant with A. flavus colonization is its potential to secrete mycotoxins, of which the most prominent is aflatoxin. Temperature, water activity (aw) and carbon dioxide (CO2) are three environmental factors shown to influence the fungus-plant interaction, which are predicted to undergo significant changes in the next century. In this study, we used RNA sequencing to better understand the transcriptomic response of the fungus to aw, temperature, and elevated CO2 levels. We demonstrate that aflatoxin (AFB1) production on maize grain was altered by water availability, temperature and CO2. RNA-Sequencing data indicated that several genes, and in particular those involved in the biosynthesis of secondary metabolites, exhibit different responses to water availability or temperature stress depending on the atmospheric CO2 content. Other gene categories affected by CO2 levels alone (350 ppm vs. 1000 ppm at 30 °C/0.99 aw), included amino acid metabolism and folate biosynthesis. Finally, we identified two gene networks significantly influenced by changes in CO2 levels that contain several genes related to cellular replication and transcription. These results demonstrate that changes in atmospheric CO2 under climate change scenarios greatly influences the response of A. flavus to water and temperature when colonizing maize grain.Item Open Access Carbon dioxide production as an indicator of Aspergillus flavus colonisation and aflatoxins/cyclopiazonic acid contamination in shelled peanuts stored under different interacting abiotic factors(Elsevier, 2019-10-24) Garcia-Cela, Esther; Gari Sanchez, F. J.; Sulyok, Michael; Verheecke-Vaessen, Carol; Medina, Angel; Kruska, Rudolf; Magan, NareshAspergillus flavus is the main xerophylic species colonising stored peanuts resulting in contamination with aflatoxins (AFs) and cyclopiazonic acid (CPA). This study evaluated the relationship between storage of shelled peanuts under interacting abiotic conditions on (a) temporal respiration (R) and cumulative CO2 production, (b) dry matter losses (DMLs) and (c) aflatoxin B1 (AFB1) and CPA accumulation. Both naturally contaminated peanuts and those inoculated with A. flavus were stored for 7-days under different water activities (aw; 0.77–0.95) and temperatures (20–35°C). There was an increase in the temporal CO2 production rates in wetter and warmer conditions, with the highest respiration at 0.95 aw + A. flavus inoculum at 30°C (2474 mg CO2kg−1h−1). The DMLs were modelled to produce contour maps of the environmental conditions resulting in maximum/minimum losses. Maximum mycotoxin contamination was always at 0.95 aw although optimal temperatures were 25-30°C for AFs and 30-35°C for CPA. These results showed a correlation between CO2 production and mycotoxin accumulation. They also provide valuable information for the creation of a database focused on the development of a post-harvest decision support system to determine the relative risks of contamination with these mycotoxins in stored shelled peanuts.Item Open Access Climate change, food security and mycotoxins: do we know enough?(Elsevier, 2017-05-18) Medina-Vayá, Ángel; Akbar, Asya; Baazeem, Alaa; Rodriguez, Alicia; Magan, NareshClimate change (CC) scenarios are predicted to have significant effects on the security of staple commodities. A key component of this impact is the infection of such crops by mycotoxigenic moulds and contamination with mycotoxins. The impacts of CC on mycotoxigenic fungi requires examination of the impacts of the three-way interactions between elevated CO2 (350–400 vs 650–1200 ppm), temperature increases (+2–5 °C) and drought stress on growth/mycotoxin production by key spoilage fungi in cereals and nuts. This review examines the available evidence on the impacts of interacting CC factors on growth and mycotoxin production by key mycotoxigenic fungi including Alternaria, Aspergillus, Fusarium and Penicillium species. Aspergillus flavus responsible for producing aflatoxin B1 (AFB1) is a class 1A carcinogen and its growth appears to be unaffected by CC factors. However, there is a significant stimulation of AFB1 production both in vitro and in vivo in maize. In contrast, studies on Aspergillus section Circumdati and Nigri species responsible for ochratoxin A contamination of a range of commodities and F. verticillioides and fumonisins suggest that some species are more resilient than others, especially in terms of mycotoxin production. Acclimatisation of mycotoxigenic fungal pathogens to CC factors may result in increased disease and perhaps mycotoxin contamination of staple cereals. Predictive modelling approaches to help identify regions where maximum impact may occur in terms of infection by mycotoxigenic fungi and toxin contamination of staple crops is hindered by the lack of reliable inputs on effects of the interacting CC factors. The present available knowledge is discussed in the context of the resilience of staple food chains and the impact that interacting CC factors may have on the availability of food in the future.Item Open Access Comparative growth inhibition of bread spoilage fungi by different preservative concentrations using a rapid turbidimetric assay system(Frontiers, 2021-06-08) Valle Garcia, Marcelo; Garcia-Cela, Esther; Magan, Naresh; Venturini Copetti, Marina; Medina, AngelBread and intermediate moisture bakery products are mainly spoiled by yeasts and filamentous fungi. The inoculum load and preservation system used determines their shelf life. To extend the shelf life of such commodities, the use of chemical preservatives is the most common way to try and control the initiation of mold spoilage of bread. This study has utilized a rapid turbidimetric assay system (Bioscreen C) to examine the temporal efficacy of calcium propionate (CP) and potassium sorbate (PS) for controlling the growth of important bread spoilage fungi. The objectives were to compare the temporal growth of strains of three important spoilage fungi Hyphopichia burtonii (HB17), Paecilomyces variotii (PV11), and Penicillium roqueforti (PR06) isolated from visibly molded bread to (a) different concentrations of CP and PS (0–128 mM), (b) temperatures (25°C, 30°C), (c) water activity (aw; 0.95, 0.97), and (d) pH (5.0, 5.5). All three abiotic factors, pH, aw, and temperature, and preservative concentrations influenced the relative growth of the species examined. In general, PS was more effective than CP in inhibiting the growth of the strains of these three species. In addition, the Time to Detection (TTD) for the efficacy of the preservatives under the interacting abiotic factors was compared. The strain of Paecilomyces variotii (PV10) was the most tolerant to the preservatives, with the shortest TTD values for both preservatives. P. roqueforti was the most sensitive with the longest TTD values under all conditions examined. These results are discussed in the context of the evolution of resistance to food-grade preservatives by such spoilage fungi in bakery products.Item Open Access Comparison of different bead-beating RNA extraction strategies: An optimized method for filamentous fungi(Elsevier Science B.V., Amsterdam., 2012-03-01T00:00:00Z) Leite, Goncalo M.; Magan, Naresh; Medina-Vayá, ÁngelMolecular studies, especially in relation to the activity of secondary metabolite gene clusters, require the ability to extract good quality RNA from fungal biomass. This is often hindered by the cell wall structure and endogenous RNase activity in filamentous fungi. There is thus a need for rapid methods for the extraction of good quality RNA for use in microarrays and for quantitative PCR assays. The objective of this study was to examine the use of different systems for the high throughput method to extract intact RNA from filamentous fungi. Two bead beating systems with different motion patterns and speed capacities were tested in the development of the extraction protocol. They were evaluated based on the total RNA yield and overall RNA quality. The high speed bead beating with glass beads associated with an automated purification method gave more than three times higher total RNA yields with less than a quarter of the amount of mycelium required. Furthermore the integrity and overall quality was conserved, with RNA Quality Indicator (RQI) numbers consistently >7.5. This method also reduced cross contamination risks and kept RNA handling to a minimum while still being capable of multiple sample processing, reducing the time required to obtain RNA from filamentous fungi.Item Open Access Comparison of different physical methods and preservatives for control of Fusarium proliferatum rot in garlic(MDPI, 2022-12-15) La Placa, Laura; Giorni, Paola; Mondani, Letizia; Magan, Naresh; Battilani, PaolaDry rot is an emerging issue for garlic production worldwide and Fusarium proliferatum is its major causal agent. Since the disease is seed-transmitted, sowing healthy cloves is crucial. In this study, some disinfection strategies were tested on garlic seeds, including steam, dry heat, chemical disinfectants and gaseous ozone (O3). Steam reduced the Colony Forming Units·g−1 (CFUs·g−1) by up to 92% in garlic seeds, but, at the same time, it affected their germination (−36%). Similarly, hydrogen peroxide (H2O2) and peracetic acid (C2H4O3) reduced the CFUs·g−1 by up to 83%; however, these methods also severely impaired germination (−40%). Dry heat did not negatively impact germination, but fungal contamination was not significantly reduced. The most promising strategy was gaseous O3 treatment; it decreased CFUs·g−1 by up to 96%, without causing any reduction of germination. The treatments applied were partially effective because the fungus is predominantly located in the outer layer of the seed, although it is also found in the inner portions. Some of these treatments can contribute to garlic protection from seed-borne pathogens and possibly reduce the occurrence of garlic dry rot.Item Open Access Comparison of dry matter losses and aflatoxin B1 contamination of paddy and brown rice stored naturally or after inoculation with Aspergillus flavus at different environmental conditions(Elsevier, 2017-07-07) Martin, Sara; Medina-Vayá, Ángel; Magan, NareshThe 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.Item Open Access Comparison of growth and aflatoxin B1 production profiles of Aspergillus flavus strains on conventional and isogenic GM-maize-based nutritional matrices(Elsevier, 2021-10-30) Gasperini, Alessandra M.; Medina, Angel; Magan, NareshMaize grown in both North and South America are now predominantly genetically modified (GM) cultivars with some resistance to herbicide, pesticide, or both. There is little information on the relative colonisation and aflatoxin B1 (AFB1) production with maize meal-based nutritional matrices based on kernels of non-GM maize and isogenic GM-ones by strains of Aspergillus flavus. The objectives were to examine the effect of interacting conditions of temperature (25–35 °C) and water availability (0.99–0.90 water activity, aw) on (a) mycelial growth, (b) AFB1 production and (c) develop contour maps of optimum and marginal conditions of these parameters for four strains of A. flavus on three different non-GM and isogenic GM-maize based nutritional media. The growth of the four strains of A. flavus (three aflatoxigenic; one non-aflatoxigenic) was relatively similar in relation to the temperature × aw conditions examined on both non-GM and GM-based matrices. Optimum growth overall was at 30–35 °C and 0.99 aw for all four strains. Under water stress (0.90 aw) growth was optimum at 35 °C. Statistically: non-GM, GM cultivars, temperature and aw all significantly affected growth rates. For AFB1 production, all single and interacting factors were statistically significant except for non-GM × GM cultivar. In conclusion, colonisation of GM- and non-GM nutritional sources was similar for the different A. flavus strains examined. The contour maps will be very useful for understanding the ecological niches for both toxigenic and non-toxigenic strains in the context of the competitive exclusion of those producing aflatoxins.Item Open Access Comparison of multiple mycotoxins in harvested maize samples in three years (2018-2020) in four continents(Taylor and Francis, 2022-01-19) Raj, Jog; Farkaš, Hunor; Jakovčević, Zdenka; Medina, Angel; Magan, Naresh; Čepela, Robert; Vasiljević, MarkoThis study has examined the pattern of mycotoxin contamination of maize destined for animal feed in different global regions over a period of 3 years (2018–2020) with up to 1000+ samples analysed in each year. Overall, >75% of samples in each of the survey years were contaminated with multiple mycotoxins regardless of the global region (Europe, Africa, Asia, South Americas countries). Using LC-MS/MS, it was possible to quantify the relative contamination present in the samples in each year from the different regions of eight different mycotoxins including aflatoxin B1 (AFB1), ochratoxin A (OTA) deoxynivalenol (DON), fumonisin B1 (FB1) and B2, zearalenone (ZEA), T-2 and HT-2 toxins. The trends in mycotoxin contamination showed that there was a consistent contamination with DON in the 3 sampling years in all four regions. Interestingly, AFB1 contamination was prevalent in all regions in 2018, but more predominant in Europe and in 2019. In contrast, in 2020 it was found to be the major contaminant in Africa only. However, FB1 contamination of maize which was prevalent in Europe in 2018, became more prevalent in Asia and LATAM countries in 2019 and even in African maize in 2020. Comparisons of contamination with different mycotoxins in each of the years globally showed significant differences for AFB1, FB1, DON and ZEA between the different years. These results are discussed in relation to the trends of contamination of maize with mixtures of mycotoxins and the implication for their control in this key commodity used as an important ingredient in animal feed.