Browsing by Author "Verheecke-Vaessen, Carol"
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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 Embargo Actinobacteria: an underestimated source of potential microbial biocontrol agents against fusarium-related diseases in cultivated crops(Springer, 2023-07-28) Djemouai, Nadjette; Meklat, Atika; Yekkour, Amine; Verheecke-Vaessen, CarolCrop infections with Fusarium spp. and associated mycotoxins have devastating economic implications and are a significant limitation to fifteen of the seventeen United Nations Sustainable Development Goals. Therefore, it is necessary to develop innovative strategies to reduce or eliminate diseases caused by Fusarium spp. in cultivated crops. Management of Fusarium-related diseases using beneficial microorganisms is a promising and eco-friendly solution. In this context, actinobacteria are considered one of the best candidates because of their multifunctional properties, which consist in the reduction of Fusarium spp. diseases and the promotion of plant growth. Actinobacteria represent a large Gram-positive bacterial phylum that is widespread in different environments, especially in soil. They are principally known for their versatile capacity to produce biologically active molecules with potential applications in different fields, including agriculture. As Fusarium species threaten diverse crops worldwide and cause global yield and quality losses that can negatively impact food security, the use of actinobacteria for plant diseases management is gaining interest as a safer and more potent alternative to ineffective synthetic-based pesticides. Thus, this work aims to review and highlight the importance of actinobacteria as microbial biocontrol agents and discuss the current research on Fusarium spp. diseases with a focus on species that threaten economically important crops, such as F. graminearum, F. oxysporum, F. solani, and F. culmorum. This review also presents actinobacteria-based products that have been patented for Fusarium disease control. We aim to provide valuable data, guidance and advancements for future research on microbial biocontrol against Fusarium-related diseases. In addition, this review emphasizes the need for further research about the impact of actinobacteria-Fusarium interactions on mycotoxin production. It also highlights the need of in-depth understanding microbial biocontrol mechanisms to ensure safe and sustainable crop production worldwide.Item Open Access Assessment of agricultural practices by Ethiopian women farmers: existence of gender disparities in access to mycotoxins training(Wageningen Academic, 2023-01-01) Cervini, Carla; Abegaz, B.; Mohammed, A.; Elias, R.; Medina, Angel; Gebre, K.; Verheecke-Vaessen, CarolEthiopia is one of the countries with the lowest gender-equality performance in sub-Saharan Africa being ranked 121/134 in terms of the magnitude and scope of gender disparities by the United Nations Women’s Organisation. Within the farming communities, women represent 70% of the labour force, but they are neglected from accessing training events run by Ethiopian Universities (e.g. Haramaya University). A survey to assess the existence of gender disparities among Ethiopian women farmers with respect to agricultural labour and mycotoxins knowledge was conducted on three hundred and forty-nine women from the Oromia and Amhara regions. A higher illiteracy rate was found in women compared to men from both Oromia and Amhara regions. Women played a key role in agricultural activities while having limited access to modern technologies compared to their male counterparts. Women were mainly responsible for sorting spoiled crops. Especially in Amhara, these were intended for home consumption, representing a serious health risk for local people. Overall, women from Amhara were more aware than women from Oromia about what mycotoxins are (e.g. aflatoxins), their impact and risk of occurrence in crops. Women in Amhara were also more intended to act towards mycotoxins in the future compared to women from Oromia. Only 0.24% of women have previously attended a training on mycotoxins. The radio seemed to be the most efficient way to deliver training to Ethiopian women farmers from these regions. Mycotoxins trainings were the second option of choice by all women surveyed. Such findings clearly stated the existence of gender inequality in the two Ethiopian regions considered. Empower women’s knowledge about mycotoxins will not only benefit agricultural income and the national economy, but it will also provide women the recognition they equally deserve alongside their male counterparts in future agricultural training programs and interventions.Item Embargo Biocontrol activities of yeasts or lactic acid bacteria isolated from Robusta coffee against Aspergillus carbonarius growth and ochratoxin A production in vitro(Elsevier, 2024-03-01) López Rodrígue, Claudia; Strub, Caroline; Fontana, Angélique; Verheecke-Vaessen, Carol; Durand, Noël; Beugré, Corinne; Guehi, Tagro; Medina, Angel; Schorr-Galindo, SabineBiocontrol Agents (BCAs) can be an eco-friendly alternative to fungicides to reduce the contamination with mycotoxigenic fungi on coffee. In the present study, different strains of bacteria and yeasts were isolated from Ivorian Robusta coffee. Their ability to reduce fungal growth and Ochratoxin A (OTA) production during their confrontation against Aspergillus carbonarius was screened on solid media. Some strains were able to reduce growth and OTA production by 85 % and 90 % and were molecularly identified as two yeasts, Rhodosporidiobolus ruineniae and Meyerozyma caribbica. Subsequent tests on liquid media with A. carbonarius or solely with OTA revealed adhesion of R. ruineniae to the mycelium of A. carbonarius through Scanning Electron Microscopy, and an OTA adsorption efficiency of 50 %. For M. caribbica potential degradation of OTA after 24 h incubation was observed. Both yeasts could be potential BCAs good candidates for Ivorian Robusta coffee protection against A. carbonarius and OTA contamination.Item Open Access Biological activities of Streptomyces sp. Bts40 isolated from the rhizosphere of Artemisia herba-alba Asso(Universitatea din Oradea, 2022-01-04) Djemouai, Nadjette; Meklat, Atika; Gaceb-Terrak, Rabéa; Oulad Hadj Youcef, Khadidja; Nacer, Asma; Mokrane, Salim; Bouras, Noureddine; Verheecke-Vaessen, CarolActinobacteria isolated from the rhizosphere of plants are of interest as they produce a diverse range of molecules, such as antibiotics and enzymes. This study investigates the antibacterial activity, plant growth-promoting (PGP) abilities as well as the production of extracellular enzymes by the actinobacterial strain BTS40. This strain was isolated from the rhizospheric soil of the medicinal plant Artemisia herba-alba Asso that was naturally grown in a semi-arid environment. Morphological characteristics showed that the strain BTS40 belongs to the genus Streptomyces. Analysis of BTS40’s 16S rRNA gene sequence showed 99.45% similarity to Streptomyces alboniger NRRL B-1832T, in the EzTaxon database. This actinobacterium showed only antibacterial activity against Gram-positive bacteria. The strain also showed potential multiple traits for plant growth promotion and hydrolysis of enzymes. Hence, this study reveals that strain BTS40 has multiple PGP traits and produces many extracellular hydrolytic enzymes.Item Open Access Biovalorisation of crude glycerol and xylose into xylitol by oleaginous yeast Yarrowia lipolytica(BMC (part of Springer Nature), 2020-06-03) Prabhu, Vinayak Ashok; Thomas, Dominic J.; Ledesma-Amaro, Rodrigo; Leeke, Gary A.; Medina, Angel; Verheecke-Vaessen, Carol; Coulon, Frederic; Agrawal, Deepti; Kumar, VinodBackground Xylitol is a commercially important chemical with multiple applications in the food and pharmaceutical industries. According to the US Department of Energy, xylitol is one of the top twelve platform chemicals that can be produced from biomass. The chemical method for xylitol synthesis is however, expensive and energy intensive. In contrast, the biological route using microbial cell factories offers a potential cost-effective alternative process. The bioprocess occurs under ambient conditions and makes use of biocatalysts and biomass which can be sourced from renewable carbon originating from a variety of cheap waste feedstocks. Result In this study, biotransformation of xylose to xylitol was investigated using Yarrowia lipolytica, an oleaginous yeast which was firstly grown on a glycerol/glucose for screening of co-substrate, followed by media optimisation in shake flask, scale up in bioreactor and downstream processing of xylitol. A two-step medium optimization was employed using central composite design and artificial neural network coupled with genetic algorithm. The yeast amassed a concentration of 53.2 g/L xylitol using pure glycerol (PG) and xylose with a bioconversion yield of 0.97 g/g. Similar results were obtained when PG was substituted with crude glycerol (CG) from the biodiesel industry (titer: 50.5 g/L; yield: 0.92 g/g). Even when xylose from sugarcane bagasse hydrolysate was used as opposed to pure xylose, a xylitol yield of 0.54 g/g was achieved. Xylitol was successfully crystallized from PG/xylose and CG/xylose fermentation broths with a recovery of 39.5 and 35.3%, respectively. Conclusion To the best of the author’s knowledge, this study demonstrates for the first time the potential of using Y. lipolytica as a microbial cell factory for xylitol synthesis from inexpensive feedstocks. The results obtained are competitive with other xylitol producing organisms.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 Essential oils modulate gene expression and ochratoxin a production in Aspergillus carbonarius(MDPI, 2016-08-19) El Khoury, Rachelle; Atoui, Ali; Verheecke-Vaessen, Carol; Maroun, Richard; El Khoury, André; Mathieu, FlorenceOchratoxin A (OTA) is a mycotoxin, mainly produced on grapes by Aspergillus carbonarius, that causes massive health problems for humans. This study aims to reduce the occurrence of OTA by using the ten following essential oils (E.Os): fennel, cardamom, anise, chamomile, celery, cinnamon, thyme, taramira, oregano and rosemary at 1 µL/mL and 5 µL/mL for each E.O. As a matter of fact, their effects on the OTA production and the growth of A. carbonarius S402 cultures were evaluated, after four days at 28 C on a Synthetic Grape Medium (SGM). Results showed that A. carbonarius growth was reduced up to 100%, when cultured with the E.Os of cinnamon, taramira, and oregano at both concentrations and the thyme at 5 µL/mL. As for the other six E.Os, their effect on A. carbonarius growth was insignificant, but highly important on the OTA production. Interestingly, the fennel E.O at 5 µL/mL reduced the OTA production up to 88.9% compared to the control, with only 13.8% of fungal growth reduction. We further investigated the effect of these E.Os on the expression levels of the genes responsible for the OTA biosynthesis (acOTApks and acOTAnrps along with the acpks gene) as well as the two regulatory genes laeA and vea, using the quantitative Reverse Transcription-Polymerase Chain Reaction (qRT-PCR) method. The results revealed that these six E.Os reduced the expression of the five studied genes, where the ackps was downregulated by 99.2% (the highest downregulation in this study) with 5 µL/mL of fennel E.O. As for the acOTApks, acOTAnrps, veA and laeA, their reduction levels ranged between 10% and 96% depending on the nature of the E.O and its concentration in the medium.Item Open Access Genome‐wide association mapping of Fusarium langsethiae infection and mycotoxin accumulation in oat (Avena sativa L.)(Wiley, 2020-06-08) Isidro-Sánchez, Julio; Cusack, Kane D’Arcy; Verheecke-Vaessen, Carol; Kahla, Amal; Bekele, Wubishet; Doohan, Fiona; Magan, Naresh; Medina, AngelFusarium langsethiae is a symptomless pathogen of oat panicles that produces T‐2 and HT‐2 mycotoxins, two of the most potent trichothecenes produced by Fusarium fungi in cereals. In the last few years, the levels of these mycotoxin in oat grain has increased and the European commission have already recommended a maximum level for of 1000 μg kg−1 for unprocessed oat for human consumption. The optimal and most sustainable way of combating infection and mycotoxin contamination is by releasing resistant oat varieties. Here the objective was to determine if we could identify any genomic loci associated with either the accumulation of F. langsethiae DNA or mycotoxins in the grain. In each of two years, field trials were conducted wherein 190 spring oat varieties were inoculated with a mixture of three isolate of the pathogen. Mycotoxins were quantified using liquid chromatography–tandem mass spectrometry. Varieties were genotyped using 16,863 genotyping by sequencing markers. Genome‐wide association studies associated 5 SNPs in the linkage group Mr06 with T‐2 + HT‐2 mycotoxin accumulation. Markers were highly correlated, and a single QTL was identified. The marker avgbs_6K_95238.1 mapped within genes showing similarity to lipase, lipase‐like or lipase precursor mRNA sequences and zinc‐finger proteins. These regions have previously been shown to confer a significant increase in resistance to Fusarium species.Item Open Access Growth kinetic parameters and prediction of growth and zearalenone and deoxynivalenol production boundaries by three Fusarium asiaticum strains isolated from wheat(MDPI, 2022-10-25) Garcia-Cela, Esther; Verheecke-Vaessen, Carol; Ósk-Jónsdóttir, Inga; Lawson, Rita; Magan, NareshFusarium species can cause head blight of cereals worldwide. This is accompanied by impacts on yield and contamination of grains with mycotoxins. Regulations, with maximum limits, exist for the relevant Fusarium mycotoxins (e.g., type A and B trichothecenes, zearalenone and fumonisins). There is interest in a better understanding of the effect of key interacting abiotic factors which determine colonization and mycotoxin production in small grain cereals. Thus, this study examined the ecophysiological relationship between temperature and water availability (10–35 °C; water activity, aw, 0.87–0.98) on growth and production of Fusarium mycotoxins (zearelenone, ZEA; deoxynivalenol, DON; 3-acetyl deoxynivalenol, 3-Ac-DON and nivalenol, NIV) by three strains of F. asiaticum, a head blight pathogen isolated from China and becoming important in other global regions. These were carried out on simulated wheat-based matrices that identified the optimum (25 °C/0.98 aw) and marginal boundary conditions for growth (35 °C/0.90 aw) for all three strains. Contrarily, different mycotoxigenic profiles were observed between strains (p < 0.05). Four mycotoxins assessed were produced at 30 °C while cold temperature inhibited the production of NIV and ZEA, which were never detected at <20 and <15 °C, respectively. Optimal mycotoxin production conditions varied for each toxin with ZEA production which was best at 30 °C/0.93–0.95 aw, DON, 3-Ac-DON and NIV which was 0.98 aw/20–30 °C. Probabilistic models were used to predict growth and regulated mycotoxin production by the strains of F. asiaticum. This study will be beneficial in the development mitigation strategies for control of pre- and post-harvest colonization of cereals and mycotoxin contamination by this Fusarium species in cereals.Item Open Access Harvest and post-harvest handling practices associated with Fumonisin B1 Contamination in maize (Zea mays L.): dietary exposure and risk characterization in Eastern Ethiopia(Springer, 2022-09-30) Mohammed, Abdi; Seid, Awol; Terefe, Habtamu; Cervini, Carla; Verheecke-Vaessen, CarolMaize is the main staple food crop in the eastern part of Ethiopia. However, maize loss is a major issue due to fungal contamination especially at the post-harvest stage owing to inadequate handling practices. This study aimed to assess post-harvest handling and awareness against fungal development and fumonisin B1 (FB1) in maize and to calculate risk exposures of FB1. A total of 197 maize samples (grain and flour) were collected from five districts (Haramaya, Kersa, Meta, Oda Bultum, and Tullo). FB1 was detected using LC–MS/MS qTRAP. Exposure assessment was done based on the maize consumption rate per day in Ethiopia for different age groups (infants, children, and adults). Risk characterization depends on the margin of exposure (MoE) combined with the lower confidence limit of the benchmark dose level (BMDL). About 81% of farmers were not physically separating undamaged maize ears with damaged from either birds or fungi. Moreover, 100% were not using improved storage material. In storage samples, FB1 was detected as high as 1058 μg/kg ± 234 in the Kersa district while a minimum of 22.60 μg/kg ± 5.27 in Meta. In flour samples, the maximum FB1 (327 μg/kg) was detected from the Oda Bultum district. The maximum exposure of infants was estimated at Kersa (1131 µg/kg bw/day), followed by Oda Bultum (1073 µg/kg bw/day) and Haramaya (854 µg/kg bw/day). Overall, FB1 exposures ranged from 6.09 to 1131 µg/kg bw/day, which is 3 to 500 µg/kg bw/day higher than the maximum tolerable daily intake of 2 µg/kg bw/day recommended by the World Health Organization. The MoE ranged from 0.15 to 176, with infants being at higher risk than adults. The study highlights the urgent need to enhance growers’ awareness and knowledge of good post-harvest practices to reduce mycotoxin contamination in maize. Further biomarker analysis must be pursued to determine the risk exposure assessment for different age groups in these areas with a priority for the Kersa district.Item Open Access Impact of predicted climate change environmental conditions on the growth of Fusarium asiaticum strains and mycotoxins production on a wheat-based matrix(Elsevier, 2024-03-13) Cervini, Carla; Naz, Naoreen; Verheecke-Vaessen, Carol; Medina, AngelFusarium asiaticum is a predominant fungal pathogen causing Fusarium Head Blight (FHB) in wheat and barley in China and is associated with approximately £201 million in annual losses due to grains contaminated with mycotoxins. F. asiaticum produces deoxynivalenol and zearalenone whose maximum limits in cereals and cereals-derived products have been established in different countries including the EU. Few studies are available on the ecophysiological behaviour of this fungal pathogen, but nothing is known about the impact of projected climate change scenarios on its growth and mycotoxin production. Therefore, this study aimed to examine the interacting effect of i) current and increased temperature (25 vs 30 °C), ii) drought stress variation (0.98 vs 0.95 water activity; aw) and iii) existing and predicted CO2 concentrations (400 vs 1000 ppm) on fungal growth and mycotoxin production (type B trichothecenes and zearalenone) by three F. asiaticum strains (CH024b, 82, 0982) on a wheat-based matrix after 10 days of incubation. The results showed that, when exposed to increased CO2 concentration (1000 ppm) there was a significant reduction of fungal growth compared to current concentration (400 ppm) both at 25 and 30 °C, especially at 0.95 aw. The multi-mycotoxin analysis performed by LC-MS/MS qTRAP showed a significant increase of deoxynivalenol and 15-acetyldeoxynivalenol production when the CH024b strain was exposed to elevated CO2 compared to current CO2 levels. Zearalenone production by the strain 0982 was significantly stimulated by mild water stress (0.95 aw) and increased CO2 concentration (1000 ppm) regardless of the temperature. Such results highlight that intraspecies variability exist among F. asiaticum strains with some mycotoxins likely to exceed current EU legislative limits under prospected climate change conditions.Item Open Access Improvements within the peanut production chain to minimize aflatoxins contamination: an Ethiopian case study(Elsevier, 2021-10-15) Cervini, Carla; Verheecke-Vaessen, Carol; He, Tingting; Mohammed, Abdi; Magan, Naresh; Medina, AngelPeanuts (Arachis hypogaea L.) are a worldwide crop appreciated by consumers and known to have several health benefits (e.g. decreasing risk of cardiovascular diseases, lowering LDL cholesterol). Nonetheless, during the production chain, peanuts can be contaminated by mycotoxigenic fungi, especially Aspergillus flavus and A. parasiticus, responsible for contamination of this commodity with aflatoxins (AFs). Chronic exposure to these mycotoxins is known to have carcinogenic, teratogenic and immunosuppressive effects, while acute exposure can be fatal. Peanut-based products are considered the “new-gold” in Lower Middle-Income Countries (LMICs) as they are used to produce therapeutic food to fight malnutrition in children in the form of high energy bars (e.g. Plumpy’Nut®). The present study has been focused on an examination of the existing Ethiopian peanut supply chain to identify components that require improvements during pre-harvest, harvesting and post-harvest where practical and affordable intervention practices can be implemented. The lack of Good Agricultural Practices (GAPs), the use of traditional and rudimentary methods for drying and shelling peanuts, as well as inadequate storage locations are some of the main factors associated with the high contamination of Ethiopian peanuts with AFs. While control of AFs is complex, in-depth examination of existing practices has facilitated an overview of the potential for efficient and low-cost strategies to reduce and minimize these toxin risks in the Ethiopian peanut production chain. This includes the use of Drycard™ as a tool to check moisture content, the creation of homemade solar dryer and wooden nutcrackers, and the use of either the Purdue Improved Crop Storage (PICS) bags or novel bags with slow-release food-grade preservatives. The implementation of some of these approaches should ensure a safer home-grown peanut supply chain that would contribute to the growth of the economy of the country by reducing the high costs of importing such raw materials and supporting the use of local smallholder farmers peanuts. This would have significant economic benefits in the local peanut production chain.Item Open Access Incidence of mycobiota and aflatoxin B1 in Algerian feed(Inderscience, 2022-03-29) Bouti, Karima; Mimoune, Nouara Ait; Mokrane, Salim; Djemouai, Nadjette; Verheecke-Vaessen, Carol; Mathieu, Florence; Riba, AmarThe presence of fungi and aflatoxin B1 (AFB1) in 101 animal feed samples randomly collected from different vendors and factories in Algeria was investigated. For fungi, the main genera isolated were Aspergillus, Penicillium and Fusarium. Furthermore, the 459 strains of Aspergillus section Flavi were screened for their ability to produce aflatoxins and cyclopiazonic acid. 49% of the strains produced AFB1. The highest incidence of aflatoxigenic strains was recorded in maize (61%) and ground poultry feed (60%). The presence of AFB1 in feed samples was evaluated using HPLC-FLD. The obtained data showed that 36.6% of samples were contaminated in the range of 0.34 to 171.06 μg/kg. Six samples exceeded the Algerian maximum limit of 20 µg/kg for AFB1. This study highlights the potential presence of aflatoxigenic strains belonging to section Flavi and AFB1 in animal feed at post-harvest in Algeria, strategic information for the Algerian policies makers.Item Open Access Interacting climate change environmental factors effects on Fusarium langsethiae growth, expression of Tri genes and T-2/HT-2 mycotoxin production on oat-based media and in stored oats(Elsevier, 2019-05-11) Verheecke-Vaessen, Carol; Diez-Gutierrez, Lucia; Renaud, Justin; Sumarah, Mark; Medina, Angel; Magan, NareshThe objectives of this study were to investigate the impact that interactions between key climate change (CC) related environmental factors of temperature (20, 25, 30°C), water activity (aw; 0.995, 0.98) and CO2 exposure (400, 1000 ppm) may have on (a) growth, (b) gene expression of biosynthetic toxin genes (Tri5, Tri6, Tri16), and (c) phenotypic T-2/HT-2 production by Fusarium langsethiae on oat-based agar medium and in stored oats. Fungal growth was optimum at 25°C and 0.995 aw and reduced significantly at 30°C and intermediate stress (0.98 aw, elevated CO2 (1000 ppm) exposure by approx. 4-fold. Lag phases prior to growth paralleled these results with the longest lag phase in this treatment (24 hrs). On oat-based medium, the relative Tri5 gene expression was increased in elevated CO2 conditions. The expression of both the Tri6 and Tri16 genes was reduced when compared to control (20°C, 0.995 aw, 400 ppm), especially in elevated CO2 conditions. In stored oats, the Tri5 gene expression was reduced in all conditions except at 30°C, 0.98 aw, elevated CO2 where there was a significant (5.3-fold) increase. The expression of the Tri6 was slightly over-expressed in elevated CO2 and the Tri16 gene was upregulated, especially in elevated CO2 conditions. For mycotoxin production, both on oat-based medium and in stored oats the production was higher at 25°C when compared to 30°C. In stored oats, at 0.98 aw, elevated CO2 led to higher T2/HT-2 toxin production at both 25 and 30°C with a significant increase (73-fold higher) at 30°C. In elevated CO2 conditions, Tri16 (Spearman test; 0.68; p-value=0.0019) and Tri5 gene expression (Spearman test; 0.56; p-value=0.0151) were correlated with T-2+HT-2 production. Nine T-2 and HT-2 metabolites were detected by LC-MS/MS including a new dehydro T-2 toxin and the conjugate, HT-2 toxin glucuronide (in plantae). The new dehydro T-2 toxin was the most abundant metabolites and showed correlation (R2=0.8176) with T-2 production. This is the first study to examine the impact of CC factors on growth and mycotoxin production by a strain of F. langsethiae. The influence of such scenarios on relative risk of oats contamination with these toxins in relation to the food security agenda is discussed.Item Open Access Interacting climate change factors (CO2 and temperature cycles) effects on growth, secondary metabolite gene expression and phenotypic ochratoxin A production by Aspergillus carbonarius strains on a grape-based matrix(Elsevier, 2019-11-12) Cervini, Carla; Verheecke-Vaessen, Carol; Ferrara, Massimo; García-Cela, Esther; Magistà, Donato; Medina, Angel; Gallo, Antonia; Magan, Naresh; Perrone, GiancarloLittle is known on the impact that climate change (CC) may have on Aspergillus carbonarius and Ochratoxin A (OTA) contamination of grapes, especially in the Mediterranean region where in CC scenarios temperature are expected to increase by +2–5 °C and CO2 from 400 to 800/1200 ppm. This study examined the effect of (i) current and increased temperature in the alternating 11.5 h dark/12.5 h light cycle (15–28 °C vs 18–34 °C), representative of the North Apulia area, South Italy and (ii) existing and predicted CO2 concentrations (400 vs 1000 ppm), on growth, expression of biosynthetic genes (AcOTApks, AcOTAnrps, AcOTAhal, AcOTAp450, AcOTAbZIP) and regulatory genes of Velvet complex (laeA/veA/velB, “velvet complex”) involved in OTA biosynthesis and OTA phenotypic production by three strains of A. carbonarius. The experiments made on a grape-based matrix showed that elevated CO2 resulted in a general stimulation of growth and OTA production. These results were also supported by the up-regulation of both structural and regulatory genes involved in the OTA biosynthesis. Our work has shown for the first time that elevated CO2 concentration in the Mediterranean region may result in an increased risk of OTA contamination in the wine production chain.Item Open Access De novo genome assembly and functional annotation for Fusarium langsethiae(Springer, 2022-02-22) Zuo, Ya; Verheecke-Vaessen, Carol; Molitor, Corentin; Medina, Angel; Magan, Naresh; Mohareb, FadyBackground Fusarium langsethiae is a T-2 and HT-2 mycotoxins producing species firstly characterised in 2004. It is commonly isolated from oats in Northern Europe. T-2 and HT-2 mycotoxins exhibit immunological and haemotological effects in animal health mainly through inhibition of protein, RNA and DNA synthesis. The development of a high-quality and comprehensively annotated assembly for this species is therefore essential in providing the molecular understanding and the mechanism of T-2 and HT-2 biosynthesis in F. langsethiae to help develop effective control strategies. Results The F. langsethiae assembly was produced using PacBio long reads, which were then assembled independently using Canu, SMARTdenovo and Flye. A total of 19,336 coding genes were identified using RNA-Seq informed ab-initio gene prediction. Finally, predicting genes were annotated using the basic local alignment search tool (BLAST) against the NCBI non-redundant (NR) genome database and protein hits were annotated using InterProScan. Genes with blast hits were functionally annotated with Gene Ontology. Conclusions We developed a high-quality genome assembly of a total length of 59 Mb and N50 of 3.51 Mb. Raw sequence reads and assembled genome is publicly available and can be downloaded from: GenBank under the accession JAFFKB000000000.Item Open Access The "-omics" contributions to the understanding of mycotoxin production under diverse environmental conditions(Elsevier, 2018-08-24) Garcia-Cela, Esther; Verheecke-Vaessen, Carol; Magan, Naresh; Medina-Vayá, ÁngelExtreme environmental changes and fluctuations mainly driven by climate change will have a profound effect on natural food contaminants. Among these contaminants mycotoxins will be very important due the high adaptability of the producing fungal genera to the forecasted conditions. The availability of modern, high through-put –omic techniques, including genomics, transcriptomics, metagenomics, proteomics and metabolomics has facilitated a rapid expansion of data on the biology of mycotoxigenic fungi. This has facilitated a significant increase in our knowledge of the biological, biochemical and biophysical molecular processes regulating the production of mycotoxins, and the adaptation of these fungi to environmental stresses. In this paper we highlight recent advances where -omics approaches have been used and where they have contributed to the knowledge on how mycotoxigenic fungi adapt to diverse interacting environmental conditions and their relationship with phenotypic toxin production. We also highlight potential future directions where these approaches can be effectively utilised for the development of minimisation strategies in the context of expected climate change scenarios and the food security agenda.Item Open Access Polyphasic characterization of Aspergillus section Flavi isolated from animal feeds in Algeria(Wiley, 2019-12-13) Bouti, Karima; Verheecke-Vaessen, Carol; Mokrane, Salim; Meklat, Atika; Djemouai, Nadjette; Sabaou, Nasserdine; Mathieu, Florence; Riba, AmarIn Algeria, little information is available on the population structure of Aspergillus section Flavi in raw materials and resultant animal feeds. A total of 172 isolates belonging to Aspergillus section Flavi were recovered from 57 animal feeds and identified on the basis of macro and micro‐morphological characters, mycotoxin production and genetic relatedness. For the molecular analysis, sequencing of the calmodulin gene (CaM) and the internal transcribed spacer (ITS) regions were performed for representative isolates. Four distinct morphotypes were distinguished: Aspergillus flavus (78.5%), Aspergillus tamarii (19.2%), Aspergillus parasiticus (1.7%), and Aspergillus alliaceus (0.6%). All A. flavus isolates were of the L type and no correlation between sclerotia production and aflatoxigenicity was observed. Our results showed that 68% of the A. flavus strains produced aflatoxins B (AFB), and 72.7% were cyclopiazonic acid (CPA) producers. The three isolates of A. parasiticus were able to produce AFB and aflatoxins G but not CPA whereas, all the strains of A. tamarii produced only CPA. The obtained results revealed the presence of different species of Aspergillus section Flavi, among which were aflatoxin producers. This study provides evidence useful for considerations in aflatoxin control strategies.Item Open Access Proof of concept: could snake venoms be a potential source of bioactive compounds for control of mould growth and mycotoxin production(Wiley, 2020-06-05) Verheecke-Vaessen, Carol; Monte, J.; Garcia-Cela, Esther; Magan, Naresh; Medina, AngelThe objective was to screen 10 snake venoms for their efficacy to control growth and mycotoxin production by important mycotoxigenic fungi including Aspergillus flavus , Aspergillus westerdijkiae , Penicillium verrucosum , Fusarium graminearum and F. langsethiae . The Bioscreen C rapid assay system was used. The venoms from the Viperidae snake family delayed growth of some of the test fungi, especially F. graminearum and F. langsethiae and sometimes A. flavus . Some were also able to reduce mycotoxin production. The two most potent crude snake venoms (Naja nigricollis and N. siamensis; 41 and 43 fractions, respectively) were further fractionated and 83/84 of these fractions were able to reduce mycotoxin production by >90% in two of the mycotoxigenic fungi examined. This study suggests that there may be significant potential for the identification of novel fungistatic/fungicidal bioactive compounds as preservatives of raw and processed food commodities post‐harvest from such snake venoms.