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International audience; Scientific communication is facilitated by a data-driven, scientifically sound taxonomy that considers the end-user's needs and established successful practice. Previously (Geiser et al. 2013; Phytopathology 103:400-408. 2013), the Fusarium community voiced near unanimous support for a concept of Fusarium that represented a clade comprising all agriculturally and clinically important Fusarium species, including the F. solani Species Complex (FSSC). Subsequently, this concept was challenged by one research group (Lombard et al. 2015 Studies in Mycology 80: 189-245) who proposed dividing Fusarium into seven genera, including the FSSC as the genus Neocosmospora, with subsequent justification based on claims that the Geiser et al. (2013) concept of Fusarium is polyphyletic (Sandoval-Denis et al. 2018; Persoonia 41:109-129). Here we test this claim, and provide a phylogeny based on exonic nucleotide sequences of 19 orthologous protein-coding genes that strongly support the monophyly of Fusarium including the FSSC. We reassert the practical and scientific argument in support of a Fusarium that includes the FSSC and several other basal lineages, consistent with the longstanding use of this name among plant pathologists, medical mycologists, quarantine officials, regulatory agencies, students and researchers with a stake in its taxonomy. In recognition of this monophyly, 40 species recently described as Neocosmospora were recombined in Fusarium, and nine others were renamed Fusarium. Here the global Fusarium community voices strong support for the inclusion of the FSSC in Fusarium, as it remains the best scientific, nomenclatural and practical taxonomic option available.

Ergot alkaloids (EAs) are mycotoxins mainly produced by the fungus Claviceps purpurea. EAs are known to affect the nervous system and to be vasoconstrictors in humans and animals. This work presents recent advances in swine and dairy feeds regarding 11 major EAs, namely ergometrine, ergosine, ergotamine, ergocornine, ergocryptine, ergocristine, ergosinine, ergotaminine, ergocorninine, ergocryptinine, and ergocristinine. A reliable, sensitive, and accurate multiple mycotoxin method, based on extraction with a Mycosep 150 multifunctional column prior to analysis using UHPLC-MS/MS, was validated using samples of swine feed (100) and dairy feed (100) for the 11 targeted EAs. Based on the obtained validation results, this method showed good performance recovery and inter-day and intra-day precision that are in accordance with standard criteria to ensure reliable occurrence data on EA contaminants. More than 49% of the swine feed samples were contaminated with EAs, especially ergocryptine(-ine) (40%) and ergosine (-ine) and ergotamine (-ine) (37%). However, many of the 11 EAs were not detectable in any swine feed samples. In addition, there were contaminated (positive) dairy feed samples, especially for ergocryptine (-ine) (50%), ergosine (-ine) (48%), ergotamine (-ine), and ergocristine (-ine) (49%). The mycotoxin levels in the feed samples in this study almost complied with the European Union regulations.

Four taxa of the Lamiaceae family growing in Apulia (Clinopodium suaveolens, Satureja montana subsp. montana, Thymbra capitata, and Salvia fruticosa subsp. thomasii) that had not been previously studied for their potential use in the food sector, were analyzed for their essential oils (EOs) composition and antioxidant and antimicrobial properties against some microorganisms, isolated from bread and bakery products, including molds (Aspergillus niger, Penicillium roqueforti) and spore-forming bacteria (Bacillus amyloliquefaciens and Bacillus subtilis). Two different sites were considered for each plant species, and the strongest antimicrobial EOs, which were active against all of the microorganisms tested, were those from one S. montana subsp. montana sample (Sm2) and both T. capitata EOs (Tc1 and Tc2) with Minimal Inhibitory Concentration (MIC) values ranging between 0.093% and 0.375% (v/v) against molds, while higher values were registered for bacteria (0.75–1%). In particular, the biological activity of EOs from T. capitata and S. montana subsp. montana was maybe due to the high amount of thymol and carvacrol, which were also responsible for the highest antioxidant activity. S. fruticosa subsp. thomasii EOs had different chemical profiles but showed only a slight antibacterial effect and no antifungal activity. C. suaveolens showed no significant changes between EOs with an interesting antifungal activity (MIC 0.093%÷0.187% v/v), which may be due to the presence of pulegone. These plant species can be considered as promising sources of bioactive compounds to be exploited as biopreservatives in bread and bakery products mainly considering the low concentration needed to inhibit microorganism’s growth.

Yeasts are eukaryotic microorganisms which have a long history in the biotechnology of food production, as they have been used since centuries in bread-making or in the production of alcoholic beverages such as wines or beers. Relative to this importance, a lot of research has been devoted to the study of yeasts involved in making these important products. The role of yeasts in other fermentations in association with other microorganisms - mainly lactic acid bacteria - has been relatively less studied, and often it is not clear if yeasts occurring in such fermentations are contaminants with no role in the fermentation, spoilage microorganisms or whether they actually serve a technological or functional purpose. Some knowledge is available for yeasts used as starter cultures in fermented raw sausages or in the production of acid curd cheeses. This review aimed to summarize the current knowledge on the taxonomy, the presence and potential functional or technological roles of yeasts in traditional fermented plant, dairy, fish and meat fermentations.

The effect of oxalic acid (OA) treatment on visual properties and bioactive compounds of two green and one purple cultivars of fresh asparagus was investigated during twelve storage days at 5 °C. Cold storage and OA treatment positively affected the overall appearance of the investigated cultivars. Cut-end dehydration increased, all along the storage period, in all cultivars but, the negative effect of the storage, clear on control samples, was mitigated by OA. The most represented compounds in Grande and Vegalim cultivars were: quercetin rutinoside, feruloyl quinic acid and cumaroyl quinic acid. Cyanidin glucosyl rutinoside, cyaniding rutinoside and peonidin rutinoside were identified in Purple Passion cultivar. The bioactive compounds seemed to be affected by storage but not by OA treatment. The sensor-biosensor system indicated that the antioxidant activity is negatively affected by storage but not by OA. The decrease of antioxidant activity coincided with the reduction of ascorbic acid levels in all the cultivars.

Sugarcane is an important crop in Southern Iran for agri-food, energy, and pharmaceutical industries. Among the pathogens that colonize sugarcane, mycotoxigenic Fusarium species are reason of serious concern for both their pathogenicity on plants and ability to produce harmful mycotoxins to humans and animals. We studied 104 Fusarium strains, selected within a wider Fusarium set isolated from sugarcane in Southern Iran, for molecular identification, phylogeny and mycotoxin analyses. Most of Fusarium strains belonged to Fusarium fujikuroi Species Complex (FFSC) and identified mainly as F. proliferatum, at minor extent as F. sacchari, and rarely as F. thapsinum, and F. verticillioides. Moreover, 14 strains identified as FFSC could not be assigned to any known species, although they were phylogenetically closely related to F. andiyazi, likely representing a new phylogenetic species. A subset of FFSC strains were analyzed for in vitro production of fumonisins (FBs), beauvericin (BEA), and enniatins (ENNs). Fusarium proliferatum strains produced FBs at high amount, and, at a lesser extent, BEA, and ENNs; F.sacchari produced only BEA and B ENNs at very low level; Fusarium sp. strains produced only B ENNs. The paper provides new insights on the genetic diversity of Fusarium species and their mycotoxin profile occurring on sugarcane in Iran.

The prevalence of mycotoxins is often increased by the climatic conditions prevailing in tropical regions. Reports have revealed the contamination of mycotoxins in some types of vegetable oil. However, vegetable oil is one of the essential ingredients used in food preparation. Thus, this study determined the occurrence of multi-mycotoxins in six types of vegetable oils commercially available in Thailand to assess the consumer health risk. In total, 300 vegetable oil samples (olive oil, palm oil, soybean oil, corn oil, sunflower oil, and rice bran oil) collected from various markets in Thailand were analyzed for the presence of nine mycotoxins, namely, aflatoxin B1 (AFB1), aflatoxin B2 (AFB2), aflatoxin G1 (AFG1), aflatoxin G2 (AFG2), beauvericin (BEA), ochratoxin A (OTA), zearalenone (ZEA), fumonisin B1 (FB1), and fumonisin B2 (FB2) using a quick, easy, cheap, effective, rugged, and safe (QuEChERS)-based procedure and a triple quadrupole mass spectrometer equipped with an electrospray ionization source. The incidences of mycotoxin contamination varied among the different types of oil samples. AFB1, AFB2, ZEA, FB1, and FB2 were most frequently found in contaminated samples. AFB2, BEA, ZEA, FB1, and FB2 contaminated olive oil samples, whereas AFB1, AFB2, AFG2, and OTA contaminated palm oil samples. AFB1, AFB2, and ZEA were found in soybean oils, whereas ZEA, FB1, and FB2 contaminated corn oil samples. AFB1 and AFG1 contaminated sunflower oil samples, whereas AFB1, AFB2, AFG1, and OTA were detected in rice bran oil samples. However, the contamination levels of the analyzed mycotoxins were below the regulatory limits.

During the last decade, there have been many advances in research and technology that have greatly contributed to expanded capabilities and knowledge in detection and measurement, characterization, biosynthesis, and management of mycotoxins in maize. MycoKey, an EU‐funded Horizon 2020 project, was established to advance knowledge and technology transfer around the globe to address mycotoxin impacts in key food and feed chains. MycoKey included several working groups comprising international experts in different fields of mycotoxicology. The MycoKey Maize Working Group recently convened to gather information and strategize for the development and implementation of solutions to the maize mycotoxin problem in light of current and emerging technologies. This feature summarizes the Maize WG discussion and recommendations for addressing mycotoxin problems in maize. Discussions focused on aflatoxins, deoxynivalenol, fumonisins, and zearalenone, which are the most widespread and persistently important mycotoxins in maize. Although regional differences were recognized, there was consensus about many of the priorities for research and effective management strategies. For preharvest management, genetic resistance and selecting adapted maize genotypes, along with insect management, were among the most fruitful strategies identified across the mycotoxin groups. For postharvest management, the most important practices included timely harvest, rapid grain drying, grain cleaning, and carefully managed storage conditions. Remediation practices such as optical sorting, density separation, milling, and chemical detoxification were also suggested. Future research and communication priorities included advanced breeding technologies, development of risk assessment tools, and the development and dissemination of regionally relevant management guidelines.

Aspergillus-produced mycotoxins can enter the feed and food chain at many points in both preharvest and post-harvest. Although current climate changes seem to speed up the world-wide spread of mycotoxigenic fungi including the Aspergilli and also facilitate the production of these harmful secondary metabolites the factors governing these disadvantageous global processes are only partly understood or even have remained completely hidden until now. This Research Topic summarizes our knowledge on Aspergillus-derived mycotoxins especially focusing on three major areas of on-going research: (i) toxicological, medical, veterinary aspects, prevalence, detection, risk assessment, control strategies, (ii) ecology and biological control of mycotoxigenic Aspergilli in the fields, and (iii) pre-harvest and post-harvest management of mycotoxigenic Aspergilli and their mycotoxin production. We hope that the wealth of information generously provided by the Aspergillus mycotoxin research community will help the hard work of all those experts, who are active in this important field, and the papers collected here will be instructive and illuminating readings for students and the public as well. Aspergillus-produced mycotoxins can enter the feed and food chain at many points in both pre- harvest and post-harvest. Although current climate changes seem to speed up the world-wide spread of mycotoxigenic fungi including the Aspergilli and also facilitate the production of these harmful secondary metabolites the factors governing these disadvantageous global processes are only partly understood or even have remained completely hidden until now. This Research Topic summarizes our knowledge on Aspergillus-derived mycotoxins especially focusing on three major areas of on-going research: (i) toxicological, medical, veterinary aspects, prevalence, detection, risk assessment, control strategies, (ii) ecology and biological control of mycotoxigenic Aspergilli in the fields, and (iii) pre-harvest and post-harvest management of mycotoxigenic Aspergilli and their mycotoxin production. We hope that the wealth of information generously provided by the Aspergillus mycotoxin research community will help the hard work of all those experts, who are active in this important field, and the papers collected here will be instructive and illuminating readings for students and the public as well.

Purslane (Portulaca oleracea L.) is a wild edible plant, traditionally consumed in the Mediterranean area and recently proposed as a new ready-to-eat vegetable, it is also called the &ldquo, vegetable for long life&rdquo, because of the high contents of several healthy compounds. Although boron (B) is not considered to be essential for humans, a daily intake of about 2 mg to obtain positive effects on aging in adult men and women has been suggested. In this study, two genotypes of purslane (wild collected and commercial variety) are grown by using a hydroponic system with three boron (B) levels in the nutrient solution (NS) (0.3 mg/L&mdash, control, 3 mg/L&mdash, low level of biofortification, and 6 mg/L&mdash, high level of biofortification) in order to increase the B content in the edible parts of the plant. The crop yield, color traits, and content of glucose, fructose, total phenols, chlorophylls, carotenoids, mineral elements (Al, B, Ca, Cr, Fe, K, Mg, Mn, Na, and Zn), nitrate, and oxalate are analyzed. Independent of the genotype, the B content in edible purslane was successfully increased in comparison with the control, obtaining 1.8- to 10.7-fold higher values of B tissue concentrations by using, respectively, 3 and 6 mg/L of B in the NS without affecting crop performances. From a nutritional point of view, the average daily intake of B could be satisfied by consuming about 75 or 48 g of purslane, grown by using 3 and 6 mg/L B level in the NS, respectively. Apart from B and Fe, the content of mineral elements in edible parts of purslane was not strongly influenced by different B levels in the NS but it was affected by genotypes. A lower sugar content was found in wild purslane grown with the highest B level. A higher content of both chlorophylls and carotenoids was found in the control but only for the commercial genotype. No differences in oxalate content were observed among B levels in the NS, while only in the case of wild genotype, we found a lower nitrate content when a B concentration of 3 mg/L was used in the NS. In conclusion, we demonstrated the possibility of using the floating hydroponic system, combined with specific B concentrations in the NS composition, as a method to calibrate the B uptake in edible parts of purslane.

Anaerobic digestion represents an interesting approach to produce biogas from organic waste materials contaminated by mycotoxins. In this study a shotgun metagenomic analysis of lab-scale bioreactors fed with mycotoxin-contaminated silage has been carried out to characterize the evolution of microbial community under the operating conditions and the key enzymatic activities responsible for mycotoxin degradation. The study was conducted at two different level of contamination for fumonisins and aflatoxin B1. After 15 days biogas production was not influenced by the presence of mycotoxins. Metagenomic analysis revealed that a high contamination rate of mycotoxins interfere with microbial diversity. Degradation of mycotoxins accounted in about 54% for aflatoxin B1 and 60% for fumonisins. The degradation activity of fumonisins resulted in the presence of partially hydrolyzed forms in both tested contamination levels. Accordingly, metagenomic functional analysis revealed the presence of two new carboxylesterase genes belonging to D. bacterium and P. bacterium putatively involved in fumonisin degradation.

Liquid chromatography coupled with high-resolution mass spectrometry (LC-HRMS) represents the most powerful metabolomics platform to investigate biological systems. Reproducible and standardized workflows allow obtaining a meaningful biological interpretation. The purpose of this study was to set up and apply an open-source workflow for LC-HRMS plant metabolomics studies. Key steps of the proposed workflow were as follows: (1) experimental design, (2) sample preparation, (3) LC-HRMS analysis, (4) data processing, (5) custom database search, (6) statistical analysis, (7) compound identification, and (8) biochemical interpretation. Its applicability was evaluated through the study of metabolomics changes of two maize recombinant inbred lines with contrasting phenotypes with respect to disease severity after Fusarium verticillioides infection of seedlings. Analysis of data from the case-control study revealed abundance change in metabolites belonging to different metabolic pathways, including two amino acids (L-tryptophan and tyrosine), five flavonoids, and three N-hydroxynnamic acid amides.

Exopolysaccharides (EPS) are known for their positive contribute to the technological properties of many foods, including bakery products. These molecules can be obtained performing piloted fermentation with lactic acid bacteria (LAB). In order to select strains able to produce EPS, a screening test in agar medium containing sucrose, fructose or glucose as carbohydrate source was performed on 21 LAB strains. Results allowed to select 8 Weissella cibaria, 2 Weissella confusa and 2 Leuconostoc spp. strains as EPS producers only in the presence of sucrose. A further screening in liquid medium enriched with sucrose (10%) (mMRS_S) indicated the W. cibaria strain C43-11 as the higher EPS producer. The selected strain was used to develop liquid sourdoughs (LSs) with dough yield (DY) 500, fermented for 15 h and based on wheat flour and wheat gluten or pseudocereals (quinoa or amaranth) in ratio 1:1, in the presence or not of sucrose at 3% (w/w, LS weight), in comparison to Lactobacillus plantarum ITM21B, a strain not producing EPS in mMRS_S. Results indicated that the use of pseudocereals favored the EPS production. Formulations were optimized by modifying DY (500 or 250), sucrose concentration (3 or 6%) and flour ratio. LSs were characterized for the content of organic acids (lactic, acetic, phenyllactic, OH-phenyllactic), pH, TTA, EPS, viscosity, total protein degradation and protein pattern. The highest EPS production (20.79 g/Kg) and viscosity (1168 mPa s) were obtained in LS (DY 250, sucrose 6%) based on quinoa flour and started with C43-11 strain. The LS was characterized by the presence of phenyllactic and OH-phenyllactic acids, protein degradation by 51.7% and proteins in the range 14-80 kDa. In these conditions, also strain ITM21B was able to produce EPS at level of 4.61 g/Kg and to degrade proteins by 53.8% in LS based on wheat and quinoa flours (1:1) (DY250 and sucrose 3%). Therefore, results demonstrated that the use of selected conditions (flour type, DY, sucrose) can stimulate specific attributes of strains making them suitable for production of short fermented (15h) liquid sourdoughs which can be used as bread improvers.

Dehydroascorbate reductases (DHARs) are important enzymes that reconvert the dehydroascorbic acid (DHA) into ascorbic acid (ASC). They are involved in the plant response to oxidative stress, such as that induced by the mycotoxin beauvericin (BEA). Tomato plants were treated with 50 &micro, M of BEA, the main antioxidant compounds and enzymes were evaluated. DHARs were analyzed in the presence of different electron donors by native and denaturing electrophoresis as well as by western blot and mass spectrometry to identify a novel induced protein with DHAR activity. Kinetic parameters for dehydroascorbate (DHA) and glutathione (GSH) were also determined. The novel DHAR was induced after BEA treatment. It was GSH-dependent and possessed lower affinity to DHA and GSH than the classical DHARs. Interestingly, the mass spectrometry analysis of the main band appearing on sodium dodecyl sulphate polyacrylamide gel electrophoresis (SDS-PAGE) revealed a chloroplast sedoheptulose 1,7-bisphosphatase, a key enzyme of the Calvin cycle, and a chloroplast mRNA-binding protein, suggesting that the DHA reducing capacity could be a side activity or the novel DHAR could be part of a protein complex. These results shed new light on the ascorbate-glutathione regulation network under oxidative stress and may represent a new way to increase the plant antioxidant defense system, plant nutraceutical value, and the health benefits of plant consumption.

Aflatoxin B1 (AFB1) is a secondary metabolite produced by some Aspergillus spp. fungi affecting many crops and feed materials. Aflatoxin M1 (AFM1), the 4-hydroxylated metabolite of AFB1, is the main AFB1-related compound present in milk, and it is categorized by the International Agency for Research on Cancer (IARC) as a &ldquo, group 1 human carcinogen&rdquo, The aim of this work was to evaluate and compare the analytical performances of two commercial immunoassays widely applied for the detection of AFM1 in milk, namely strip test immunoassay and enzyme linked immunosorbent assay (ELISA). Assay validation included samples at AFM1 levels of 25, 50, 75 ng/kg and blank samples (AFM1 &lt, 0.5 ng/kg). With respect to a screening target concentration (STC) of 50 ng/kg the two assays showed cut-off values of 37.7 ng/kg and 47.5 ng/kg for strip test and ELISA, respectively, a false suspect rate for blanks &lt, 0.1% (for both assays) and a false negative rate for samples containing AFM1 at levels higher than STC, of 0.4% (for both assays). The intermediate precision (RSDip) was &lt, 32% for the strip test and &lt, 15% for the ELISA. Method verification through long-term intra-laboratory quality control (QC) measurements confirmed the results from the validation study. Furthermore, a satisfactory correlation of the results obtained with both immunoassays and the AOAC Official Method 2000.08 was obtained for the analysis of cow milk samples naturally contaminated with AFM1 at levels within &ldquo, not detected&rdquo, (&lt, 0.5 ng/kg) and 50 ng/kg. Finally, the extension of the scope of the strip test method to goat and sheep milk was evaluated by applying the experimental design foreseen in the EU regulation.

A reliable, sensitive and accurate multiple mycotoxin method was developed for the simultaneous determination of 17 mycotoxins in swine, poultry and dairy feeds using stable isotope dilution (13C-ISTD) and (ultra)-high performance liquid chromatography-tandem mass spectrometry (HPLC-MS/MS). A simple QuEChERS-based method (quick, easy, cheap, effective, rugged and safe) was developed consisting of soaking with a solution of 1% formic acid followed by extraction with acetonitrile, clean-up with C18 sorbent and finally adding 13C-ISTD before the UHPLC-MS/MS analysis. The chromatographic condition was optimized for separation and detection of the 17 mycotoxins using gradient elution. The method&rsquo, s performance complied with the SANTE/11813/2017 standard and had mean recovery accuracies in the range 70%&ndash, 120% and precision testing of % relative standard deviation (RSD) &pound, 20%. The limit of detection and limit of quantification values ranged from 0.25 to 40.0 ng/g and 0.5 to 100.0 ng/g, respectively. Finally, the method was applied to analyze feed samples, with the results showing that fumonisins, zearalenone, aflatoxin B1 and deoxynivalenol were the most prevalent mycotoxins contaminating the feed samples.

A prototype of shipping container equipped with controlled atmosphere (CA) system (CA-prototype), was used for the simulation of the cold transport of table grapes (cv Italia). The CAprototype was realised by IFAC SPA, within the Research Project Continnova. It is equipped with a cooling system in order to work at a temperature between –20°C and +15°C. The CA management is realised through the control system and the connections with the remote application. The experimental simulation of the cold transport was realised putting 20 boxes of table grapes inside the CA-prototype (set at 20% O2 + 10% CO2 in nitrogen), other 20 in a refrigerated room using a SO2 pad inside each box, the remaining 20 in a cold room (Control) without SO2 pad for 12 days at 5°C. At the end of the simulated transport, for each postharvest solution, 10 table grapes boxes were analysed, while the remaining 10 were stored for 3 days at 20°C in air with the aim to simulate a shelf-life period. CA-prototype allows preserving table grapes visual quality, delaying browning and maintaining berry turgidity, extending the shelf-life until 12 days at 5°C. Finally, CA-prototype resulted a valid alternative to the commonly used SO2 pads, which are under constant revision because of its potential allergenic effects.

The aim of this work was to evaluate the antifungal activity in vapor phase of thymol, p-cymene, and &gamma, terpinene, the red thyme essential oil compounds (RTOCs). The Minimum Inhibitory Concentration (MIC) of RTOCs was determined against postharvest spoilage fungi of the genera Botrytis, Penicillium, Alternaria, and Monilinia, by measuring the reduction of the fungal biomass after exposure for 72 h at 25 &deg, C. Thymol showed the lowest MIC (7.0 &micro, g/L), followed by &gamma, terpinene (28.4 &micro, g/L) and p-cymene (40.0 &micro, g/L). In the case of P. digitatum ITEM 9569, resistant to commercial RTO, a better evaluation of interactions among RTOCs was performed using the checkerboard assay and the calculation of the Fractional Inhibitory Concentration Index (FICI). During incubation, changes in the RTOCs concentration were measured by GC-MS analysis. A synergistic effect between thymol (0.013 &plusmn, 0.003 L/L) and &gamma, terpinene (0.990 &plusmn, 0.030 L/L) (FICI = 0.50) in binary combinations, and between p-cymene (0.700 &plusmn, 0.010 L/L) and &gamma, terpinene (0.290 &plusmn, 0.010 L/L) in presence of thymol (0.008 &plusmn, 0.001 L/L) (FICI = 0.19), in ternary combinations was found. The synergistic effect against the strain P. digitatum ITEM 9569 suggests that different combinations among RTOCs could be defined to control fungal strains causing different food spoilage phenomena.

Ligninolytic enzymes from white-rot fungi, such as laccase (Lac) and Mn-peroxidase (MnP), are able to degrade aflatoxin B1 (AFB1), the most harmful among the known mycotoxins. The high cost of purification of these enzymes has limited their implementation into practical technologies. Every year, tons of spent mushroom substrate (SMS) are produced as a by-product of edible mushroom cultivation, such as Pleurotus spp., and disposed at a cost for farmers. SMS may still bea source of ligninolytic enzymes useful for AFB1 degradation. The in vitro AFB1-degradative activity of an SMS crude extract (SMSE) was investigated. Results show that: (1) in SMSE, high Lac activity (4 U g&minus, 1dry matter) and low MnP activity (0.4 U g&minus, 1dry matter) were present, (2) after 1 d of incubation at 25 &deg, C, the SMSE was able to degrade more than 50% of AFB1, whereas after 3 and 7 d of incubation, the percentage of degradation reached the values of 75% and 90%, respectively, (3) with increasing pH values, the degradation percentage increased, reaching 90% after 3 d at pH 8. Based on these results, SMS proved to be a suitable source of AFB1 degrading enzymes and the use of SMSE to detoxify AFB1 contaminated commodities appears conceivable.

Maize is the principal staple food/feed crop exposed to mycotoxins, and the co-occurrence of multiple mycotoxins and their metabolites has been well documented. This review presents the infection cycle, ecology, and plant-pathogen interactions of Aspergillus and Fusarium species in maize, and current knowledge on maize chain management to mitigate the occurrence of aflatoxins and fumonisins. Preventive actions include at pre-harvest, as part of cropping systems, at harvest, and at postharvest, through storage, processing, and detoxification to minimize consumer exposure. Preventive actions in the field have been recognized as efficient for reducing the entrance of mycotoxins into production chains. Biological control of Aspergillus flavus has been recognized to minimize contamination with aflatoxins. Post-harvest maize grain management is also crucial to complete preventive actions, and has been made mandatory in government food and feed legislation., This review was prepared as part of MYCHIF EFSA project (GP/EFSA/AFSCO/2016/01). Roberta Palumbo carried out this work within the PhD school Agrisystem of Università Cattolica del Sacro Cuore, Italy. This study was supported by the Portuguese Foundation for Science and Technology (FCT) under the scope of the strategic funding of UID/BIO/04469 unit and COMPETE 2020 (POCI-01-0145-FEDER-006684) and BioTecNorte operation (NORTE-01-0145-FEDER-000004) funded by the European Regional Development Fund under the scope of Norte2020 - Programa Operacional Regional do Norte. This paper was critically reviewed in collaboration with MycoKey project (Horizon 2020, Grant Agreement No. 678781)., info:eu-repo/semantics/publishedVersion

Ochratoxin A (OTA) occurrence in grapes is caused by black Aspergilli (Aspergillus carbonarius followed by A. niger) vineyards contamination. It depends on climatic conditions, geographical regions, damage by insects, and grape varieties. Good agricultural practices, pesticides, and fungicides seem adequate to manage the problem during low OTA risk vintages, but the development of new strategies is always encouraged, especially when an extremely favourable condition occurs in the vineyard. Electrolysed oxidising water (EOW) has become an interesting alternative to chemicals in agriculture, mainly during the post-harvest phase. This study tested the fungicidal efficacy of EOW generated by potassium chloride, in vitro, on black Aspergilli conidia, and detached grape berries infected by A. carbonarius. Then, during field trials on Primitivo cv vineyard treated with EOW, A. carbonarius contamination, and OTA levels were compared with Switch® fungicide treatment (0.8 g/l). Black Aspergilli conidia were killed on plate assay after 2 min of treatment by EOW containing >0.4 g/l of active chlorine. EOW (0.6 g/l active chlorine) treatment reduced the rate of A. carbonarius infections in vitro of about 87–92% on detached berries and, more than half in the field trials, although Switch® showed better performance. A significant reduction in the OTA concentration was observed for the EOW and Switch® treatments in vitro (92% and 96%, respectively), while in the field trials, although the average decrease in OTA was recorded in the treated grapes, it was not statistically significant. These results highlighted that EOW could be considered effective, as a substitute for fungicides, to reduce the contamination of A. carbonarius and OTA on grapes.

Table grapes (cv. Italia) were stored at 5 °C in air or in modified atmosphere with low O 2 (1%) and low (0.03–10%) or high (>20%) CO 2 . Volatile organic compounds (VOCs) associated with quality (respiration rate, acetaldehyde and ethanol, water status of rachis) and sensory parameters (visual quality, rachis browning, characteristic odour and flavour, off-odour and off-flavour) were evaluated. Low CO 2 preserved the quality and sensory parameters, whereas high CO 2 caused a fermentative metabolism. A total of 126 VOCs were identified by HS-SPME/GC–MS analysis. Principal component analysis (PCA) identified ( Z )-2-hexenal and (E,Z)-2,4-hexadienal as potential marker of freshness; whereas, acetaldehyde, ethanol, 1-butanol, 3-methyl-1-butanol, 1-octanol and 1-nonanol were correlated to fermentation processes. Partial least-square regression analysis (PLSR) was used to identify (R 2 = 0.95) the main predictors of off-odour as ethanol and ethyl acetate (+) and hexanal and β-linalool (−).

Non-destructive evaluation of vegetables by Computer Vision Systems (CVSs) makes possible to check their quality level in an objective and consistent way along the whole supply chain up to the final users. CVSs have been proven to be successful when applied to unpackaged products. The proposed approach aimed to enable this analysis on packaged fresh-cut lettuce with minimum constraints on the acquisition phase and without any care to flatten the surface of the bag facing the camera. A deep-learning architecture, based on Convolutional Neural Networks (CNNs), was used to identify regions of the image where the vegetable was visible with minimum colour distortions due to packaging. To meaningfully assess the performance of the system, each lettuce's sample was acquired both through packaging material and without packaging material. The image analysis was applied to both the resulting images to automatically grade their quality level. The results showed that the performance loss due to the presence of packaging is negligible (83% instead of 86%) and that the proposed system can be used to monitor the quality level of fresh-cut lettuce regardless of packaging at all the critical check points along the supply chain.

As determined by the Intergovernmental Panel on Climate Change, warming of the climate system is unequivocal and has been associated with rising sea levels, diminished amounts of ice and snow and increasing oceanic and atmospheric temperatures. Such climate changes have a significant impact on stages and rates of toxigenic fungi development and can modify host-resistance and host-pathogen interactions, influencing deeply also the conditions for mycotoxin production that vary for each individual pathogen. Moreover, the new combinations mycotoxins/host plants/geographical areas are arising to the attention of the scientific community and require new diagnostic tools and deeper knowledge of both biology and genetics of toxigenic fungi. In this review, it is underlined that an extension of the aflatoxin contamination risk in maize in South and Central-Europe is highly likely in the next 30 years, due to favorable climatic conditions to the growth of Aspergillus flavus. Moreover, the mycotoxigenic Fusarium species profile on wheat in Europe is in continuous change in Northern, Central and Southern-Europe with, in particular, a worrisome growing contamination of F. graminearum in the Central and Northern Europe.

Barley can be contaminated with a wide range of fungal secondary metabolites, including various mycotoxins that reduce the quality and safety of raw materials as well as cause economic losses. A survey was conducted for the crop seasons 2016 and 2017 to analyse fungal metabolites, including mycotoxins, in grain and straw samples of barley, which originated from fields across Switzerland. In total, 253 grain and 237 straw samples were analysed by LC-MS/MS detecting 87 and 86 fungal metabolites, respectively, which are reported to be produced by Fusarium, Alternaria, Claviceps, Aspergillus, Penicillium and other genera. None of the grain samples exceeded the permitted limits of mycotoxins set by the European Commission. With regard to straw, three and six samples exceeded the guidance levels set for raw grains for deoxynivalenol and the sum of T-2 and HT-2, respectively. Nevertheless, some samples contained high concentrations of unregulated fungal metabolites, e.g. enniatins, infectopyron, zinniol and rubellin D. This was more frequently observed in straw and, to a lesser extent, in grains, suggesting that the presence of fungal metabolites in straw material should not be neglected. Our study demonstrated that both grain and straw matrices of barley represent large pools of various fungal secondary metabolites, most of them with undetermined toxicity. Hence, future studies should focus on the toxicology of the predominant fungal metabolites that occurred at elevated concentrations as well as the health impact of co-occurrence of toxins primarily with metabolites that revealed strong correlations.

The healthy consumers make a strong pressure to natural products that can prevent the chronic diseases and improve the general health status, and therefore an important aspect that have to be considered is the safe level of the nutraceuticals. This study reports the occurrence of Ochratoxin A (OTA) and associated fungal contamination in 35 samples of dried vine fruits imported in the European community potentially used for the development of new nutraceutical supplements. High pressure liquid chromatography analysis identified 18 samples as contaminated by OTA with an average level of 2.6 μg/kg. OTA was measured in 4 samples of currants (mean value of 6.6 μg/kg) and 13 samples of raisins (mean value of 1.4 μg/kg). In one sample of currants and one of raisins from Turkey OTA exceeded the limits set by European Commission of 10 μg/kg, being contaminated with 12.61 and 15.99 μg/kg, respectively. All the positive samples were confirmed by Orbitrap Q Exactive through their molecular weight and the corresponding fragmentation. The worldwide consumption of dried vine fruits contributed to OTA exposure in several group of consumers. In particular, considering the potential nutraceutical approach, this consumption may be represent a severe risk for healthy consumers that consider these products like healthy and salutistic for their contents in antioxidants, flavonoids, and polyphenols. Data reported in this study confirmed the need to regularly monitor mycotoxin levels in these food products and optimize the process of fruits drying in order to reduce the development of toxigenic molds.

Cakile maritima is a typical halophyte of the Mediterranean coasts. In addition to its ecological and industrial properties, C. maritima has antiscorbutic, diuretic and purgative roles in folk remedies. This plant is infected by different fungal species, mainly belonging to Alternaria genus. Two-hundred Alternaria strains were collected from four different pedo-climatic areas in Tunisia, from C. maritima fresh plant tissues showing symptoms of Alternaria infection. Phylogenetic analyses of 79 representative Alternaria strains, were carried out using multi-locus gene sequencing. All the strains clustered in the Alternaria Section: 47 strains had high homology with A. alternata reference strain, 13 grouped with A. arborescens reference strain, 12 grouped with A. mali reference strain, and seven strains were not well defined with A. mali as their closest species. In vitro production of tenuazonic acid (TA), alternariol (AOH), alternariol-monomethyl ether (AME), and altenuene (ALT) was evaluated. Approx. 68% of strains simultaneously produced AOH, AME and TA. Only two A. alternata and one A. mali strains were ALT producing. Pathogenicity tests on leaves of C. maritima were carried out with 41 representative strains. Alternaria arborescens showed the greatest pathogenicity compared to A. alternata and A. mali, although no statistically significant differences in pathogenicity were observed. This is the first study on Tunisian populations of Alternaria species isolated from the extremophile C. maritima.

Fusarium proliferatum causes diverse diseases of many economically important plants. The fungus produces several mycotoxins of which the fumonisins are the most toxic. Currently, deletion of key genes for mycotoxin biosynthesis is a laborious and time-consuming procedure. We developed a novel CRISPR/Cas9-based genome-editing tool for the direct delivery of preassembled Cas9 ribonucleoproteins into protoplasts of F. proliferatum. Our CRISPR–Cas9 system couples a site-specific double-strand DNA break mediated by two Cas9 ribonucleoproteins with microhomology recombination requiring only 50-bp regions flanking the target gene. This system reduces the risk of off-target mutations and minimizes the risk of altering any gene adjacent to the target region. We used this tool to delete a polyketide synthase gene (FUM1) required for fumonisin biosynthesis. The mutants generated are no longer able to produce fumonisins, confirming the key role of FUM1 in fumonisin biosynthesis. Our CRISPR-Cas9 system is an important new tool for genetic studies of Fusarium.

Fusarium fujikuroi species complex (FFSC) species are commonly encountered infecting rice, but knowledge of the diversity and toxigenic potential of the species is lacking in Brazil, the largest rice-producing country outside Asia. One hundred FFSC isolates obtained from national rice were identified using morphology and phylogeny of TEF, CAL and TUB genes. Eight previously known and one novel Fusarium species were identified. Three species accounted for around 60% of the strains: F. fujikuroi (n = 23), F. proliferatum (n = 22) and F. verticillioides (n = 16). The less frequent species were F. volatile (n = 8), F. anthophilum (n = 6), F. pseudocircinatum (n = 4), F. sterilihyphosum (n = 2) and F. begoniae (n = 1). The novel Fusarium species was represented by 18 isolates. All species produced at least one of the analyzed mycotoxins [beauvericin (BEA), fumonisins (FBs), moniliformin (MON) and enniatins (ENNs)]. BEA was produced by all species but F. verticillioides. The FBs (mainly FB1) were produced mostly by F. fujikuroi, F. proliferatum and F. verticillioides. F. begoniae and F. verticillioides did not produce ENNs and F. sterilihyphosum and F. begoniae did not produce MON, while the other species produced MON and ENNs. Our results add new knowledge of the diversity, geographical distribution and host range of FFSC species.

Tailored foods are specifically suitable for target groups of people with particular nutritional needs. Although most research on tailored foods has been focused on increasing the nutrient content in plant tissues (biofortification), in populations with specific physiological conditions, it is recommended to reduce the uptake of specific nutrients in order to improve their health. People affected by chronic kidney disease (CKD) must limit their consumption of vegetables because of the generally high potassium (K) content in the edible parts. This study aimed to define an appropriate production technique for two baby leaf vegetables, spinach (Spinacia oleracea L.) and Swiss chard (Beta vulgaris L. ssp. vulgaris), with reduced K tissue content, minimizing the negative effects on their crop performance and overall nutritional quality. Plants were grown in a hydroponic floating system. The K concentration in the nutrient solution (NS) was reduced from 200 mg/L (K200, the concentration usually used for growing baby leaf vegetables in hydroponic conditions) to 50 mg/L over the entire growing cycle (K50) or only during the seven days before harvest (K50-7d). The reduction of K in the NS resulted in a significant decrease of K tissue content in both species (32% for K50 and 10% for K50-7d, on average), while it did not, in general, compromise the crop performance and quality traits or the bioaccessibility of K, magnesium, and calcium. The production of reduced-potassium leafy vegetables is a feasible tailored nutrition approach for CKD patients in order to take advantage of the positive effects of vegetable consumption on health without excessively increasing potassium intake.

Aflatoxin B-1 (AfB(1)) is a carcinogenic mycotoxin that contaminates food and feed worldwide. We determined the AfB(1)-adsorption capability of non-viable Pleurotus eryngii mycelium, an edible fungus, as a potential means for removal of AfB(1) from contaminated solutions. Lyophilized mycelium was produced and made enzymatically inert by sterilization at high temperatures. The material thus obtained was characterized by scanning electron microscopy with regard to the morpho-structural properties of the mycotoxin-adsorbing surfaces. The active surfaces appeared rough and sponge-like. The AfB(1)-mycelium system reached equilibrium at 37 degrees C, 30 min, and pH 5-7, conditions that are compatible with the gastro-intestinal system of animals. The system remained stable for 48 h at room temperature, at pH 3, pH 7, and pH 7.4. A thermodynamic study of the process showed that this is a spontaneous and physical adsorption process, with a maximum of 85 +/- 13% of removal efficiency of AfB1 by P. eryngii mycelium. These results suggest that biosorbent materials obtained from the mycelium of the mushroom P. eryngii could be used as a low-cost and effective feed additive for AfB(1) detoxification.

Contamination with multiple mycotoxins was found in rice and barley. BEA, DAS, ZEA, and aflatoxins were the mycotoxins most frequently found in samples. The assessed mycotoxin exposure does not represent a health risk for consumers.

Fusarium incarnatum-equiseti species complex (FIESC) is commonly detected in Brazilian rice, but knowledge of the species limits and their toxigenic potential is lacking. Seventy strains morphologically identified as FIESC-like, isolated from the major rice-growing regions of Brazil, were subjected to sequencing of EF-1? gene. Among them, 18 strains were selected and analyzed for their RPB2 gene sequences. Nine phylogenetic species were identified, among which eight matched the previously reported FIESC 4 (F. lacertarum), 6, 16, 17 (F. pernambucanum), 20 (F. caatingaense), 24, 26 and 29. One new phylogenetic species was identified, and named FIESC 38. Five strains formed new singleton lineages. The most dominant species were FIESC 26 (22/70 strains) and FIESC 38 (21/70), the newly identified species. The incarnatum morphotype was dominant (10 phylogenetic species) over the equiseti (4 species). Among 46 strains selected to represent all species, only 16 strains produced detectable levels of mycotoxins in vitro. FIESC 26 produced ZEA and FIESC 38 produced both ZEA and DON. ZEA was produced by nine isolates of three other species, among which few isolates produced trichothecenes: DON (5/46), NIV (3/46), 4-ANIV (2/46), 15-ADON (1/46) and 3-ADON (1/46). The T-2 and HT-2 mycotoxins were not detected. Our results contribute novel information on species limits and mycotoxin production within cereal-infecting FIESC in the southern hemisphere and provide baseline data for further exploring morphological differences among the species.

Background The Fusarium incarnatum-equiseti species complex (FIESC) comprises 33 phylogenetically distinct species that have been recovered from diverse biological sources, but have been most often isolated from agricultural plants and soils. Collectively, members of FIESC can produce diverse mycotoxins. However, because the species diversity of FIESC has been recognized only recently, the potential of species to cause mycotoxin contamination of crop plants is unclear. In this study, therefore, we used comparative genomics to investigate the distribution of and variation in genes and gene clusters responsible for the synthesis of mycotoxins and other secondary metabolites (SMs) in FIESC. Results We examined genomes of 13 members of FIESC that were selected based primarily on their phylogenetic diversity and/or occurrence on crops. The presence and absence of SM biosynthetic gene clusters varied markedly among the genomes. For example, the trichothecene mycotoxin as well as the carotenoid and fusarubin pigment clusters were present in all genomes examined, whereas the enniatin, fusarin, and zearalenone mycotoxin clusters were present in only some genomes. Some clusters exhibited discontinuous patterns of distribution in that their presence and absence was not correlated with the phylogenetic relationships of species. We also found evidence that cluster loss and horizontal gene transfer have contributed to such distribution patterns. For example, a combination of multiple phylogenetic analyses suggest that five NRPS and seven PKS genes were introduced into FIESC from other Fusarium lineages. Conclusion Our results suggest that although the portion of the genome devoted to SM biosynthesis has remained similar during the evolutionary diversification of FIESC, the ability to produce SMs could be affected by the different distribution of related functional and complete gene clusters. Electronic supplementary material The online version of this article (10.1186/s12864-019-5567-7) contains supplementary material, which is available to authorized users.

Rapid screening methods are currently recognized as a strategic tool for mycotoxin issues management. Specific guidelines for validation and verification of mycotoxin screening methods are set in the Commission Regulation (EU) No 2014/519. This regulation establishes that the “aim of the validation is to demonstrate the fitness-for-purpose of the screening method„ and focuses the entire validation procedure on determining specific cut-off values ensuring a maximum rate of false negative results of 5%. In addition, the assessment of the rate of false suspect results is addressed. With regard to rapid test-kits, ‘fitness-for-purpose’ includes not only the criteria more commonly considered when discussing laboratory-based methods (specificity, accuracy, and precision), but also more “practical„ parameters such as speed and ease of implementation in a new operational environment. The latter means demonstrating under local conditions that performance parameters, as established during the validation, can be achieved by first time users. This goal can be achieved through “method verification„. The aim of the present study was to verify the fitness-for-purpose of mycotoxin screening methods when applied by first time users. This was achieved in one laboratory facility via results of a training course with multiple technicians attending. The verification study was organized similarly to a collaborative exercise and involved two groups comprising of 10 technicians each that used the methods for the first time. Different screening methods were applied for deoxynivalenol (DON) in wheat, which was mainly Enzyme Linked Immunosorbent Assay (ELISA), lateral flow device (LFD), fluorescence polarization immunoassay (FPIA), and liquid chromatography-high resolution mass spectrometry (LC-HRMS). An additional verification was done for aflatoxin B1 (AFB1) in maize and wheat using LFD and LC-HRMS, respectively. The results of analyses were used to calculate intermediate precision (RSDip, covering the inter-analyst variability in preparing the analytical samples and the precision under repeatability conditions) cut-off values and false suspect rates. RSDip ranged from 6.5% to 30% for DON, and from 16% to 33% for AFB1. The highest obtained variances were associated with the AFB1 analyses due to working with much lower mass fractions. The rate of false suspect results were lower than 0.1% for all tested methods. All methods showed a fit-for-purpose method performance profile, which allowed a clear distinction of samples containing the analytes at the screening target concentration (STC) from negative control samples. Moreover, the first time users obtained method performances similar to those obtained for validation studies previously performed on the screening methods included in the training course.

Fusarium head blight (FHB) is a devastating fungal disease of small-grain cereals that causes significant yield losses and mycotoxin contamination, diminishing food and feed safety worldwide. In contrast to wheat, little is known about the agricultural practices that influence FHB and Fusarium mycotoxins in barley. Thus, a nationwide survey was conducted across Switzerland for harvest samples in 2016 and 2017, accompanied with a questionnaire to obtain information about the agricultural practices in each barley field. In total, 253 grain and 237 straw samples were analyzed. In both years, F. graminearum was the predominant Fusarium species in grains followed by F. avenaceum and F. poae. Growing maize before barley was associated with increased amount of F. graminearum DNA in grains and straw as well as with elevated concentrations of deoxynivalenol in grains of barley. On the other hand, growing pasture before barley resulted in increased incidence of F. poae and concentration of numerous mycotoxins in grains (e.g., enniatins) and straw (e.g., beauvericin). Reduced tillage practices were linked to increased incidence of F. graminearum and deoxynivalenol content in grains and straw. In contrast, conventional tillage was linked to higher incidence of F. poae. Moreover, use of spring barley was associated with decreased amount of F. graminearum DNA in grains and straw, but increased incidence of F. poae and F. avenaceum. Use of the spring variety Eunova was linked to increased concentrations of several Fusarium mycotoxins in grains (e.g., enniatins and nivalenol). Furthermore, the application of strobilurin-based fungicides was associated with higher deoxynivalenol and beauvericin contents in grains. The application of plant growth regulators was associated with increased concentration of some Fusarium mycotoxins in grains (e.g., culmorin), while absence of growth regulators application was linked to elevated concentration of some other mycotoxins (e.g., nivalenol). We conclude that individual agricultural practices can suppress some FHB causing species and reduce the associated mycotoxins, but can promote others. Hence, integrated control measures combining numerous prevention and intervention strategies should be applied for the sustainable management of mycotoxins in barley.

Mycotoxins are secondary metabolites produced by toxigenic fungi contaminating foods and feeds in pre-, post-harvest and processing, and represent a great concern worldwide, both for the economic implications and for the health of the consumers. Many environmental conditions are involved in the regulation of mycotoxin biosynthesis. Among these, light represents one of the most important signals for fungi, influencing several physiological responses such as pigmentation, sexual development and asexual conidiation, primary and secondary metabolism, including mycotoxin biosynthesis. In this review we summarise some recent findings on the effect of specific light wavelength and intensity on mycotoxin biosynthesis in the main toxigenic fungal genera. We describe the molecular mechanism underlying light perception and its involvement in the regulation of secondary metabolism, focusing on VeA, global regulator in Aspergillus nidulans, and the White-Collar proteins, key components of light response in Neurospora crassa. Light of specific wavelength and intensity exerts different effects both on growth and on toxin production depending on the fungal genus. In Penicillium spp. red (627 nm) and blue wavelengths (455-470 nm) reduce ochratoxin A (OTA) biosynthesis by modulating the level of expression of the ochratoxin polyketide synthase. Furthermore a mutual regulation between citrinin and OTA production is reported in Penicillium toxigenic species. In Aspergillus spp. the effect of light treatment is strongly dependent on the species and culture conditions. Royal blue wavelength (455 nm) of high intensity (1,700 Lux) is capable of completely inhibit fungal growth and OTA production in Aspergillus stenyii and Penicillum verrucosum. In Fusarium spp. the effect of light exposure is less effective; mycotoxin-producing species, such as Fusarium verticillioides and Fusarium proliferatum, grow better under light conditions, and fumonisin production increased. This review provides a comprehensive picture on light regulation of mycotoxin biosynthesis and discusses possible new applications of this resource in food safety.

Modified mycotoxins in foodstuffs, animal feed, and herbal medicines present considerable threats to human and animal health. The diversity and wide distribution make their presence unavoidable and difficult to control. The European Food Safety Authority also recommends setting group health-based guidance values for modified mycotoxins. Significant changes in physicochemical properties of modified mycotoxins, making their detection difficult using routine analytical methods. Given the lack of commercial analysis standards and maximum limit regulations, there remains a great challenge for accurate identification and quantitation of modified mycotoxins. This review first summarizes the latest knowledge regarding target and non-target qualitative and quantitative analysis for modified mycotoxins comprehensively. In addition, this review elucidates the major transformation mechanisms of modified mycotoxins. Overall, this work lays the foundation for improving their early detection, consumer warning, safety evaluation, and control of modified mycotoxins.

Accurate identification of fungi occurring on agrofood products is the key aspect of any prevention and pest management program, offering valuable information in leading crop health and food safety. Fungal species misidentification can dramatically impact biodiversity assessment, ecological studies, management decisions, and, concerning toxigenic fungi, health risk assessment, since they can produce a wide range of toxic secondary metabolites, referred to as mycotoxins. Since each toxigenic fungal species can have its own mycotoxin profile, a correct species identification, hereby attempted with universal DNA barcoding approach, could have a key role in mycotoxins prevention strategies. Currently, identification of single marker for species resolution in fungi has not been achieved and the analysis of multiple genes is used, with the advantage of an accurate species identification and disadvantage of difficult setting up of PCR-based diagnostic assays. In the present paper, we describe our strategy to set up a DNA-based species identification of fungal species associated with maize ear rot, combining DNA barcoding approach and species-specific primers design for PCR based assays. We have (i) investigated the appropriate molecular marker for species identification, limited to mycobiota possibly occurring on maize, identifying calmodulin gene as single taxonomically informative entity; (ii) designed 17 sets of primers for rapid identification of 14 Fusarium, 10 Aspergillus, 2 Penicillium, and 2 Talaromyces species or species groups, and finally (iii) tested specificity of the 17 set of primers, in combination with 3 additional sets previously developed.