Laccases (LCs) are multicopper oxidases that find application as versatile biocatalystsfor the green bioremediation of environmental pollutants and xenobiotics. In this study weelucidate the degrading activity of Lac2 pure enzyme form Pleurotus pulmonarius towards aflatoxinB1 (AFB1) and M1 (AFM1). LC enzyme was purified using three chromatographic steps andidentified as Lac2 through zymogram and LC-MS/MS. The degradation assays were performedin vitro at 25 °C for 72 h in buffer solution. AFB1 degradation by Lac2 direct oxidationwas 23%. Toxin degradation was also investigated in the presence of three redox mediators,(2,20-azino-bis-[3-ethylbenzothiazoline-6-sulfonic acid]) (ABTS) and two naturally-occurring phenols,acetosyringone (AS) and syringaldehyde (SA). The direct effect of the enzyme and the mediatedaction of Lac2 with redox mediators univocally proved the correlation between Lac2 activity andaflatoxins degradation. The degradation of AFB1 was enhanced by the addition of all mediators at10 mM, with AS being the most effective (90% of degradation). AFM1 was completely degraded byLac2 with all mediators at 10 mM. The novelty of this study relies on the identification of a pureenzyme as capable of degrading AFB1 and, for the first time, AFM1, and on the evidence that themechanism of an effective degradation occurs via the mediation of natural phenolic compounds.These results opened new perspective for Lac2 application in the food and feed supply chains as abiotransforming agent of AFB1 and AFM1

Milk protein crosslink through the action of enzymes represents a feasible strategy to impart new functionalities to cheese. In this work we reported the effects of a laccase mediator system (LMS) on protein crosslink and antioxidant property of curd. The crosslinking activity of a purified recombinant laccase Ery4 and a commercial enzyme preparation (cLC), with three mediators was firstly evaluated in milk and then applied before curd manufacture. Only Ery4-LMS significantly increased curd weight compared to that of the control sample. SDS-PAGE revealed that similar high molecular weight bands produced by both LMSs in milk were also retained in curds. The antioxidant activity recorded in curds with Ery4-LMS was the highest among all samples both before and after gastro-pancreatic digestion. This is the first time that a CGA-based LMS is used in manufacture of curd with improved antioxidant properties. These results open new perspectives for dairy applications.

Ochratoxin A (OTA) is a mycotoxin with a main nephrotoxic activity contaminatingseveral foodstuffs. In the present report, five soil samples collected from OTA-contaminatedvineyards were screened to isolate microorganisms able to biodegrade OTA. When cultivated inOTA-supplemented medium, OTA was converted in OTalpha by 225 bacterial isolates. To revealclonal relationships between isolates, molecular typing by using an automated rep-PCR systemwas carried out, thus showing the presence of 27 different strains (rep-PCR profiles). The16S-rRNA gene sequence analysis of an isolate representative of each rep-PCR profiles indicatedthat they belonged to five bacterial genera, namely Pseudomonas, Leclercia, Pantoea, Enterobacter, andAcinetobacter. However, further evaluation of OTA-degrading activity by the 27 strains revealedthat only Acinetobacter calcoaceticus strain 396.1 and Acinetobacter sp. strain neg1, consistentlyconserved the above property; their further characterization showed that they were able to convert82% and 91% OTA into OTalpha in six days at 24 °C, respectively. The presence of OTalpha, asthe unique OTA-degradation product was confirmed by LC-HRMS. This is the first report onOTA biodegradation by bacterial strains isolated from agricultural soils and carried out underaerobic conditions and moderate temperatures. These microorganisms might be used to detoxifyOTA-contaminated feed and could be a new source of gene(s) for the development of a novelenzymatic detoxification system.

Le micotossine sono metaboliti tossici prodotti da diversi tipi di fungi, appartenenti prevalentemente al genere Fusarium, Aspergillum e Penicillium che entrano la filiera agroalimentare attraverso la contaminazione di colture utilizzate nella produzione di alimenti e mangimi. La contaminazione da micotossine genera grande preoccupazione in tutto il mondo per le gravi implicazioni a livello economico e di salute per consumatori, sia animali che umani. Per tali motivi, in molti stati, livelli limite sono stati fissati all'interno di rigorosi quadri normativi. Tra le strategie più innovative per ridurre le micotossine l'utilizzo di enzimi si sta dimostrando molto promettente. L'invenzione descritta rivendica l'utilizzo di un enzima estratto dal fungo edibile Pleurotus eryngii per la decontaminazione di fumonisine e tossina T-2, ed il suo utilizzo per la detossificazione di alimenti e mangimi.

Ochratoxin A (OTA) is a mycotoxin denoted by a nephrotoxic activity contaminating several foodstuffs. Nowadays, the biological systems for OTA degradation to the less toxic OT? aroused great interest by the scientific community. In the present study, bacteria able to biodegrade OTA were isolated from soil samples collected in OTA-contaminated vineyards. Soil samples were collected from five vineyards of Negroamaro and Primitivo grape cultivars in Salento (Southern Italy). They were cultured in appropriate media added with OTA, mycotoxin degradation was determined by HPLC/FLC analysis and bacterial colonies were isolated by plating. Clonal relationships between isolates was assessed by using an automated rep-PCR system and then each strain was identified by 16S rRNA gene sequencing. A total of 225 bacterial isolates were able to convert OTA in OT?. The molecular analysis of the above isolates showed the presence of 27 different strains (rep-PCR profiles). The sequence analysis of the 16S-rRNA gene indicated that they belonged to five bacterial genera: Pseudomonas, Leclercia, Pantoea, Enterobacter and Acinetobacter. Additional assessment of OTA-degrading capacity of the 27 strains indicated that only the Acinetobacter calcoaceticus strain 396.1 and the A. sp. strain neg1 conserved the above property: both strains were further studied thus showing that they were able to convert 82% and 91% OTA into OT? in 6 days at 24°C, respectively. The occurrence of OT?, as the sole OTA-degradation product was established by LC-MS/MS.This is the first description on OTA biodegradation under aerobic conditions and moderate temperature by bacterial strains from agricultural soils. These microorganisms might be used to detoxify OTA contaminated feed and could be a resource for the development of a new enzymatic detoxification system.

Ochratoxin A (OTA) is a nephrotoxic and potentially carcinogenic mycotoxin produced by several species of Aspergillus and Penicillium. It is one of the major mycotoxins contaminating grain, grapes and a variety of food products, and the development of methods for reducing pre-and post-harvest contamination has drawn considerable attention. In the current study, we isolated and sequenced the genome of a novel free-living Acinetobacter strain able to degrade OTA. Biochemical studies suggest that the degradation reaction proceeds via peptide bond hydrolysis.

Pseudomonas fluorescens is a genetically and phenotypically heterogeneous species that is often reported as a spoiler of fresh foods, but it has recently been implicated in clinical infection. In this study, we sequenced the genome of P. fluorescens strain ITEM 17298, isolated from mozzarella cheese and able to cause several alterations under cold storage.

Broccoli (Brassica oleracea L. var. italica) is largely cultivated in southern Italy. It is an important source of phytonutrients, which are partially lost during postharvest storage. The aim of this work was to evaluate the overall effect of five different low-intensity light-emitting diodes (LEDs) on the quality parameters of broccoli florets over 20 d of cold storage. The level of ascorbic acid, chlorophylls, carotenoids, phenolic compounds and soluble proteins, as well as colour analysis, were evaluated. Green LED increased the chlorophyll and ascorbic acid content; white, red and yellow LEDs had a positive effect on the redox status of broccoli. Globally, only green LED had a statistically significant positive effect when considering all analysed parameters and could be proposed to prolong the shelf life of broccoli during cold storage.

Leber's hereditary optic neuropathy is a maternally inherited blinding disease caused as a result of homoplasmic point mutations in complex I subunit genes of mitochondrial DNA. It is characterized by incomplete penetrance, as only some mutation carriers become affected. Thus, the mitochondrial DNA mutation is necessary but not sufficient to cause optic neuropathy. Environmental triggers and genetic modifying factors have been considered to explain its variable penetrance. We measured the mitochondrial DNA copy number and mitochondrial mass indicators in blood cells from affected and carrier individuals, screening three large pedigrees and 39 independently collected smaller families with Leber's hereditary optic neuropathy, as well as muscle biopsies and cells isolated by laser capturing from post-mortem specimens of retina and optic nerves, the latter being the disease targets. We show that unaffected mutation carriers have a significantly higher mitochondrial DNA copy number and mitochondrial mass compared with their affected relatives and control individuals. Comparative studies of fibroblasts from affected, carriers and controls, under different paradigms of metabolic demand, show that carriers display the highest capacity for activating mitochondrial biogenesis. Therefore we postulate that the increased mitochondrial biogenesis in carriers may overcome some of the pathogenic effect of mitochondrial DNA mutations. Screening of a few selected genetic variants in candidate genes involved in mitochondrial biogenesis failed to reveal any significant association. Our study provides a valuable mechanism to explain variability of penetrance in Leber's hereditary optic neuropathy and clues for high throughput genetic screening to identify the nuclear modifying gene(s), opening an avenue to develop predictive genetic tests on disease risk and therapeutic strategies.

Members of the fungal genus Fusarium can produce numerous secondary metabolites, including the nonribosomal mycotoxins beauvericin (BEA) and enniatins (ENNs). Both mycotoxins are synthesized by the multifunctional enzyme enniatin synthetase (ESYN1) that contains both peptide synthetase and S-adenosyl-l-methionine-dependent N-methyltransferase activities. Several Fusarium species can produce ENNs, BEA or both, but the mechanism(s) enabling these differential metabolic profiles is unknown. In this study, we analyzed the primary structure of ESYN1 by sequencing esyn1 transcripts from different Fusarium species. We measured ENNs and BEA production by ultra-performance liquid chromatography coupled with photodiode array and Acquity QDa mass detector (UPLC-PDA-QDa) analyses. We predicted protein structures, compared the predictions by multivariate analysis methods and found a striking correlation between BEA/ENN-producing profiles and ESYN1 three-dimensional structures. Structural differences in the ? strand's Asn789-Ala793 and His797-Asp802 portions of the amino acid adenylation domain can be used to distinguish BEA/ENN-producing Fusarium isolates from those that produce only ENN.

This study reports the fungal and bacterial metabolites associated with natural contamination of 38 composite samples of locally processed rice from five Agro-ecological Zones of Nigeria (AEZs). The samples were evaluated for the presence of microbial metabolites with the Liquid Tandem Mass Spectrometry (LC-MS/MS) technique. Among the identified metabolites, 63 fungal and 5 bacterial metabolites were measured at varying concentrations and occurrence levels. Fusarium toxins had the highest incidence of 79% but occurred in low amounts with fumonisin B1 (FB1) having the highest percentage incidence of 39.5% and a mean of 18.52 µg/kg. Among the Aspergillus toxins, aflatoxins (AFs) occurred in 36.9% of the rice samples with aflatoxin B1 (AFB1) having the highest occurrence level of 18.4% and a mean value of 5 µg/kg. About 12 metabolites had incidence levels >50%, including beauvericin (BEA) and tryptophol which had occurrence levels of 100%. Among the emerging toxins under evaluation by international organizations such as the European Food Safety Authority (EFSA) and the Food and Agriculture Organization of the United Nations (FAO), citrinin, sterigmatocystin (STER) and beauvericin were detected with maximum values of 207, 125 and 131 ?g/kg respectively. This paper also reports the first documented evidence of the contamination of Nigerian rice by bacterial and Alternaria metabolites, nivalenol, kojic acid, STER, moniliformin, fusaric acid, fumonisin B3, citrinin, 3-nitropropionic acid, andrastin A, cytochalasins, emodin and physicon.

This study reports the distribution of fungal isolates and fungal metabolites that naturallycontaminate locally processed rice from five agro-ecological zones of Nigeria. The fungal specieswere isolated by the dilution plate technique and identified by appropriate diagnostics, whilemetabolites were determined by a liquid chromatographic tandem mass spectrometric method.Aspergillus and Penicillium species were the predominant isolates found in the rice samples whileFusarium spp. were not isolated. The mean fungal count differed significantly (p < 0.05) across thezones and ranged from 9.98 × 102 cfu g-1 in the Southern Guinea Savannah to 96.97 × 102 cfu g-1in the Derived Savannah. For 16 fungal metabolites, selected from 63 positively identified fungalmetabolites based on their concentration and spread across the zones, an occurrence map wasconstructed. The Northern Guinea Savannah recorded the highest contamination of fungalmetabolites while the Sudan Savannah zone recorded the least.

Among the woody invasive alien species colonizing non-crop areas, Ailanthus altissima (Tree ofhaven) is one of the worst and most damaging weeds. It spreads everywhere in urban, suburban andnatural areas creating dense stands and causing much damage including biodiversity loss. Its controlis very difficult because of its fast growth, its capability of regenerating as 'suckers' from buds on theroots and stem after mechanical interventions, and its production of large numbers of seeds that areeasily scattered in the environment. Within the "LIFE Alta Murgia" Project, founded by the EuropeanCommission, aimed at eradicating A. altissima from the Alta Murgia National Park using innovativeand eco-friendly control techniques, the suitability of potential microbial biological agents wereexplored. Two fungal pathogens were isolated from diseased plants, identified, tested forpathogenicity and aggressiveness, and studied for the production of bioactive metabolites. The firstresults of the studies on the two fungal agents, including the chemical and biological characterizationof the metabolites and their potential as natural herbicides, are presented.

Comparisons of draft genome sequences of three geographically distinct isolates of Fusariumfujikuroi with two recently published genome sequences from the same species suggest diverseprofiles of secondary metabolite production within F. fujikuroi. Species- and lineage-specific genes,many of which appear to exhibit expression profiles that are consistent with roles in host-pathogeninteractions and adaptation to environmental changes, are concentrated in sub-telomeric regions.These genomic compartments also exhibit distinct gene densities and compositional characteristicswith respect to other genomic partitions, and likely play a role in the generation of moleculardiversity. Our data provide additional evidence that gene duplication, divergence and differentialloss play important roles in F. fujikuroi genome evolution and suggest that hundreds of lineagespecific genes might have been acquired through horizontal gene transfer.

Genomic comparative analysis of secondary metabolite clusters in Fusarium, perspectives in the big data era.

Trichoderma atrobrunneum F.B. Rocha, P. Chaverri& W. Jaklitsch strain ITEM 908 (formerly known as T. harzianum ITEM 908), is a biocontrol strain that is being registered under the European Union regulation as an active ingredient for the production of commercial biopesticides. The strain ITEM 908 proved to be able to inhibit completely the formation of perithecia by Fusarium graminearum in dual cultures and to release in the agar medium metabolites that reduce the number of perithecia by over 70% (Altomare et al., poster presentation in this Congress). Therefore, ITEM 908 appears to be a good candidate biocontrol strain for prevention of Fusarium Head Blight (FHB) in the field by treatment of plant residues of the preceeding crop, thus reducing the primary inoculum. For the univocal characterization of this commercially valuable isolate and as a base for further studies aimed at elucidating its mechanisms of action and its physiological and molecular interactions with plants and target pathogens and pests, we sequenced the whole genome of the strain ITEM 908. The genome was sequenced on an Ion S5 platform generating around 7M bpand assembled using the Spades v5.0 software. The resulting genome sequence has an estimated size of 39,131,654 bp. The reference gene sequences of ITS and TEF1 were extracted from the genome assembly. The identification of the strain ITEM 908 at species level was obtained by phylogenetic analysis with Maximum likelihood (ML) analysis performed with one-hundred ITS-TEF1 manually concatenated dataset retrieved from sequences of Trichoderma spp. deposited in GenBank. The analysis placed ITEM 908 within the T. atrobrunneum group, close to T. afroaharzianum and T. guizhouense. Genome was annotated using the Augustus v3.1 software and 8649 genes were predicted. Approximately 3000 different pfam domains were detected and used to group genes within functional categories, including glycoside hydrolases, proteases and gene involved in stress tolerance and in secondary metabolites biosynthesis. Among these, 20 putative PKS, 8 putative NRPS and 5 putative PKS-NRPS were identified. Moreover, the secretome of T. atrobrunneum ITEM 908, consisting of 761 proteins, was in silico predicted by the software SignalP (http://www.cbs.dtu.dk/services/SignalP/), that detects the presence of the secretion signal peptide at the N-terminus in amino acid sequences of a protein.The preliminary analysis of predicted genes highlights the potential ability of T. atrobrunneum ITEM 908 to produce a broad range of enzymes involved in the biocontrol activity

So far, biocontrol agent selection has been performed mainly by time consuming in vitroconfrontation tests followed by extensive trials in greenhouse and field. An alternative approach is offered byapplication of high-throughput techniques, which allow extensive screening and comparison among strains fordesired genetic traits. In the genus Trichoderma, the past assignments of particular features or strains to one speciesneed to be reconsidered according to the recent taxonomic revisions. Here we present the genome of a biocontrolstrain formerly known as Trichoderma harzianum ITEM 908, which exhibits both growth promoting capabilities andantagonism against different fungal pathogens, including Fusarium graminearum, Rhizoctonia solani, and the rootknotnematode Meloidogyne incognita. By genomic analysis of ITEM 908 we investigated the occurrence and therelevance of genes associated to biocontrol and stress tolerance, providing a basis for future investigation aimingto unravel the complex relationships between genomic endowment and exhibited activities of this strain.Results: The MLST analysis of ITS-TEF1 concatenated datasets reclassified ITEM 908 as T. atrobrunneum, a speciesrecently described within the T. harzianum species complex and phylogenetically close to T. afroharzianum and T.guizhouense. Genomic analysis revealed the presence of a broad range of genes encoding for carbohydrate activeenzymes (CAZYmes), proteins involved in secondary metabolites production, peptaboils, epidithiodioxopiperazinesand siderophores potentially involved in parasitism, saprophytic degradation as well as in biocontrol andantagonistic activities. This abundance is comparable to other Trichoderma spp. in the T. harzianum speciescomplex, but broader than in other biocontrol species and in the species T. reesei, known for its industrialapplication in cellulase production. Comparative analysis also demonstrated similar genomic organization of majorsecondary metabolites clusters, as in other Trichoderma species.Conclusions: Reported data provide a contribution to a deeper understanding of the mode of action andidentification of activity-specific genetic markers useful for selection and improvement of biocontrol strains. Thiswork will also enlarge the availability of genomic data to perform comparative studies with the aim to correlatephenotypic differences with genetic diversity of Trichoderma species.

Advanced age is characterized by several changes, one of which is the impairment of the homeostasis of intestinal microbiota. These alterations critically influence host health and have been associated with morbidity and mortality in older adults. "Inflammaging," an age-related chronic inflammatory process, is a common trait of several conditions, including sarcopenia. Interestingly, imbalanced intestinal microbial community has been suggested to contribute to inflammaging. Changes in gut microbiota accompanying sarcopenia may be attenuated by supplementation with pre- and probiotics. Although muscle aging has been increasingly recognized as a biomarker of aging, the pathophysiology of sarcopenia is to date only partially appreciated. Due to its development in the context of the age-related inflammatory milieu, several studies favor the hypothesis of a tight connection between sarcopenia and inflammaging. However, conclusive evidence describing the signaling pathways involved has not yet been produced. Here, we review the current knowledge of the changes in intestinal microbiota that occur in advanced age with a special emphasis on findings supporting the idea of a modulation of muscle physiology through alterations in gut microbial composition and activity.

Mycotoxin contamination of staple food commodities is a relevant health and economic issue worldwide.The development of green and effective reduction strategies to counteract the contamination by multiplemycotoxins has become an urgent need. The aim of this work was to evaluate the capability of a laccase(LC) from Pleurotus eryngii and a laccase-mediator systems (LMSs) to degrade aflatoxin B1 (AFB1),fumonisin B1 (FB1), ochratoxin A (OTA), deoxynivalenol (DON), Zearalenone (ZEN) and T-2 toxin in in vitroassays. In addition, the simultaneous mycotoxin degradation capability with selected LMSs was evaluatedwith combinations of AFB1 and ZEN, and FB1 and T-2 toxin. Redox mediators were found to drasticallyincreasethe degradation efficiencies of the enzyme. AFB1, FB1, OTA, ZEN and T-2 toxin degradation by thebest performing LMS were 73%, 74%, 27%, 100% and 40%, respectively. No degradation was registered forDON. Notably, AFB1 and ZEN were simultaneously degraded by 86% and 100%, while FB1 and T-2 by 25%and 100%, respectively. LMS proved to be a promising approach to enhance degradation properties of LCenzymes and for the potential development of a multi-mycotoxin reducing method.

Aspergillus niger is a fungus able to produce the carcinogenic mycotoxins ochratoxin A (OTA) and fumonisins. We analysed the influence of light of various wavelengths on growth, conidiation, fumonisin B2 (FB2) and OTA biosynthesis by A. niger ITEM 7097. Light from both sides of the spectrum, from long (627 nm) to short wavelengths (470-455 nm), had a stimulating effect on growth, with the highest stimulation under blue (455 nm, 1,700 Lux) and short-wave blue light (390 nm). Conidiation was reduced by 40% under a short blue wavelength (455 nm, 200 Lux), but strongly promoted under light at an even shorter wavelength (390 nm), with an increase of about 200 fold in comparison to the dark. Production of FB2 and OTA was mutually regulated by light. FB2 production was promoted under light conditions: red and blue light in particular increased FB2 biosynthesis by 40%. Conversely, OTA production was greatly inhibited under red and blue light in comparison to dark incubation, with a mean reduction of about 40 fold, indicating a reverse regulation of both biosynthetic pathways. Incubation under a 390 nm wavelength repressed the production of both toxins to non-detectable levels.

Light is a very important signal for fungi since it influences many different physiological responses. We analyzed the influence of light of varying wavelength and intensity on growth, conidiation and biosynthesis of fumonisin B(1) (FB(1)), B(2) (FB(2)), and B(3) (FB(3)) by Fusarium verticillioides ITEM 10027. Wavelengths across the visible spectrum, from red (627 nm) to blue (470-455 nm), stimulated the growth and increased the fumonisin production, by up to 150 % over dark incubation. If the intensity of the 455 nm blue light increased from 200 to 1700 lx, the fumonisin biosynthesis decreased. Incubation under a short wave blue light (390 nm) showed reduced fungal growth and fumonisin production by up to 85 %. White pulsing light had no effect on growth but reduced fumonisin production to half of what observed during dark incubation. Real time reverse transcriptase (RT)-PCR was used to measure the expression level of Fum1, Fum21 and FvVE1 transcripts, which encode proteins involved in fumonisin biosynthesis. There was a significant correlation between gene expression and fumonisin production.

Ophiobolin A (O-A) is a sesterpenoid with numerous biological activities, including potential anticancer effects. Its production at an industrial level is hampered due to inability of fungus Bipolaris maydis to biosynthesise it in vitro in large amount. Among the environmental factors regulating fungal metabolism, light plays a crucial role. In this study, the use of different light wavelength (light emitting diodes (LEDs)) was evaluated to increase the O-A production. The white light allowed the highest production of the metabolite. The blue and green lights showed an inhibitory effect, reducing the production to 50%, as well as red and yellow but at a lower level. No correlation between fungal growth and metabolite production was found in relation to the light type. A novel application of LED technologies, which can be optimised to foster specific pathways and promote the production of metabolites having scientific and industrial interest was proposed.

Fumonisins are a group of mycotoxins, mainly found in maize and maize-based food and feed, associated with several diseases in animals. The impact of these toxins on the economy and health worldwide has driven several efforts to clarify the role of environmental factors that can influence fumonisin biosynthesis by the toxigenic species. We analyzed the influence of light of varying wavelength on growth and fumonisin biosynthesis by the fungus Fusarium proliferatum ITEM 1719. Light in general had a positive influence on growth, with a mean increase of the grow rate of about 40% under light exposure in comparison to the dark incubation. Wavelengths from both sides of the spectrum, from long (627 nm) to short wavelength (470-455 nm) had a stimulating effect on fumonisin biosynthesis compared to the dark incubation: fumonisins B(1) (FB(1)) and B(2) (FB(2)) production increased of about 40 fold under red, 35 fold under blue, 20 fold under royal blue, 10 fold under green, 5 fold under yellow and 3 fold under white light in comparison to the dark incubation. The transcriptional regulation of the FUM1 fumonisin biosynthesis gene was analyzed by Real time reverse transcriptase PCR quantification, revealing a correlation between fumonisin biosynthesis and gene expression. These findings show a role of light on the growth and the modulation of fumonisin biosynthesis and provide new information on the physiology of an important toxigenic maize pathogen.

Fungal culture collections are important to biologists, microbiologists,epidemiologists and others involved in health and natural sciences. Theimprovement of techniques and methods for fungal isolation and preservation hascontributed to maintain large microbial collections, which represent a rich sourceof biological sciences research, especially taxonomic, pathological and biodiversitystudies as well as industrial applications. The collection centers are responsiblefor repository reference strains and for the maintenance of these microorganisms.The ITEM Microbial Culture Collection of ISPA (Institute of Sciences and of FoodProduction) includes more than 10,000 strains belonging to various agro-food microorganismswith phytopathological and toxicological signi?cance. These microorganismsare mainly fungal pathogens belonging to toxigenic genera of Fusarium, Aspergillus, Alternaria and Penicillium. This collection is a remarkable resource inthe ?ght against mycotoxins: the increasing number of toxigenic fungi included inthis collection ensures an original genetic source for biotechnological applicationsin several ?elds of research, contributing to knowledge improvement about fungalbiology and strategies development for reducing mycotoxin contamination.

Mycotoxins are secondary metabolites produced by toxigenic fungi contaminating foods and feeds in pre-, postharvestand processing, and represent a great concern worldwide, both for the economic implications and for thehealth 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 physiologicalresponses 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 lightwavelength and intensity on mycotoxin biosynthesis in the main toxigenic fungal genera. We describe the molecularmechanism underlying light perception and its involvement in the regulation of secondary metabolism, focusing onVeA, global regulator in Aspergillus nidulans, and the White-Collar proteins, key components of light response inNeurospora crassa. Light of specific wavelength and intensity exerts different effects both on growth and on toxinproduction 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. Royalblue wavelength (455 nm) of high intensity (1,700 Lux) is capable of completely inhibit fungal growth and OTAproduction in Aspergillus stenyii and Penicillum verrucosum. In Fusarium spp. the effect of light exposure is lesseffective; mycotoxin-producing species, such as Fusarium verticillioides and Fusarium proliferatum, grow betterunder light conditions, and fumonisin production increased. This review provides a comprehensive picture onlight regulation of mycotoxin biosynthesis and discusses possible new applications of this resource in food safety.

Light is an important environmental signal which influence many different physiological responses such as pigmentation, sexual development, asexual conidiation, the circadian clock and secondary metabolism. Our studies about light regulation on metabolic pathways in fungi proved that light of specific wavelength and intensity influences fungal growth and mycotoxin production.In Fusarium proliferatum light generally had a positive influence on growth, with a mean increase of the grow rate of about 40% under light exposure in comparison to the dark. Wavelengths from both sides of the spectrum, from long (627 nm) to short wavelength (470-455 nm) had a stimulating effect on fumonisin biosynthesis: fumonisins B1 and B2 production increased of about 40 fold under red, 35 fold under blue, 20 fold under royal blue, 10 fold under green, 5 fold under yellow and 3 fold under white light in comparison to the dark incubation. In Fusarium verticillioides wavelengths from red (627 nm) to blue (470-455 nm) stimulated the growth and increased the fumonisin production, by up to 150 % over dark incubation. On the contrary if the intensity of the 455 nm blue light increased from 200 to 1700 lx, the fumonisins biosynthesis decreased. Incubation under a short wave blue light (390 nm) showed reduced fungal growth and fumonisin production by up to 85 %. White pulsing light had no effect on growth but reduced fumonisin production to half of what observed during dark incubation. In both these Fusarium spp Real time reverse transcriptase analysis showed a correlation between fumonisins biosynthesis and FUM gene expression.These findings demonstrated a role of light on the growth and the modulation of fumonisin biosynthesis and provide new information on the physiology of important toxigenic pathogens.

There is an urgent need to study in Europe the plant exposure to mycotoxin risk due to the identification of new toxigenic species, the continuous evolution of species profile on the food crops and climate changes that influence the quality of level of toxigenic fungi colonization of plant hosts. In particular, Fusarium and Aspergillus problem in Europe has enormous importance; recent epidemics in wheat in some areas of Northern and Central Europe and in grape in southern Europe have brought this problem into focus again. This concern has driven many efforts at EU level aimed to harmonize strategies for mycotoxinreduction in food and feed chain. This is the target of a large collaborative project of four-year duration (MYCORED as acronym), that was approved within the European FP7- Food, Agriculture, and Biotechnologies Work Programmes (www.mycored.eu). MYCORED aims to develop strategic solutions for reducing mycotoxin contamination in major crops. Novel methodologies, efficient handling procedures and information/dissemination, and educational strategies are considered in a context of multidisciplinary integration of know-how and technology to reduce mycotoxin exposure worldwide. The direct involvement of ICPC countries (Argentina, Egypt, Russia, South Africa) and international organizations (CIMMYT, IITA) together with strong scientific alliances with international experts and national and international societies for mycotoxicology is a strong point of the project through sharing experiences and resources from several past/ongoing mycotoxin projects in a global context. Similarly, the International Society for Mycotoxicology (ISM) (http://www.mycotoxsociety. org) aims at increasing scientific knowledge concerning biology, chemistry and any sciences/disciplines related to mycotoxins and toxigenic fungi, through membership networking, scientific meetings, symposia, discussions, technical courses and publications. In this context, it would be extremely important that MYCORED and ISM develop a network of cooperation-interaction with the whole scientific community in order to contribute to the efforts for harmonizing both research and legislation on mycotoxins.

IntroductionEnergy crisis and environmental pollution have led to an increasing interest in renewable energies. Biogas production from plant material, agricultural residual products and food wastes represents one of the most economically attractive alternative technology for biofuel production. In this regards, anaerobic digestion has been widely applied to produce methane for biofuel. Complex consortia of microorganisms are responsible for biomass degradation and biogas production involving several stages such as substrate hydrolysis, acidogenesis, acetogenesis and methanogenesis. In this sense, the next-generation high-throughput sequencing provides a powerful tool for dissecting microbial community structure and methane-producing pathways in anaerobic digestion. Here, a taxonomic and functional metagenomic analysis of microbial community residing in an industrial-scale biogas fermenter has been carried out at different steps of biogas production.MethodsSample were collected from an industrial-scale mesophilic plant, daily fed with maize silage, consisting of a three steps production taking place in a bioreactor, post-reactor and a storage tank. Total DNA was extracted from samples belonging to each stage of biogas production. Metagenomic analysis were carried out by 16S and shotgun sequencing approach. The 16S datasets were generated by sequencing the bacterial and archaeal V4 hypervariable region. Reads from 16S sequencing were aligned against SILVA ribosomal RNA sequence database by using MALT (1), while shotgun reads were aligned against NCBI-nr sequence database by using DIAMOND (2). Taxonomic binning and functional annotation were performed with MEGAN 6 software (3). ResultsOver 14.5 million high quality reads (about 3.4 gigabases) were generated on the Ion Torrent S5 Sequencing System. About 2.4 and 3 million reads were assigned for 16S and shotgun approach, respectively. Although the average number of assigned taxa for 16S analysis was considerably lower than shotgun analysis, the overall taxa distribution resulting from both sequencing strategies was conserved. In detail, metagenomic analysis revealed that the superkingdom of Bacteria was dominant (~93%) along the production steps, whereas Archaea were less represented (~4%). Within Bacteria the most abundant phyla were Firmicutes, mostly represented by Clostridia, followed by Bacteroidetes, Synergistetes and Proteobacteria. Within the superkingdom of Archaea, only microorganisms belonging to the phylum of Euryarchaeota were detected. Within Euryarchaeota the dominant genera were Methanosarcina and Methanoculleus, notably to be key microorganisms involved in methanogenesis. Data showed that during biogas production steps the abundance of Methanosarcina genus decreased from bioreactor to storage tank, with a simultaneous increase of Methanoculleus genus. Functional analysis of assigned reads also supported a shift from acetotrophic methanogens to hydrogenotrophic methanogens.

Biogas production represents one of the most economically attractive alternative technology for biofuel production from renewable resources. Generally, biogas plants are fed with agricultural residual products and food wastes, but the rising up of agricultural products contaminated by mycotoxins, such as maize silage not suitable for animal feeding, has pointed the question on the possibility to use this agricultural productfor biogas production. In this regards, a preliminary metagenomic analysis of microbial community residing in a mesophilic industrial-scale biogas fermenter, daily fed with contaminated maize silage, has been carried out to characterize the evolution of microbial community under the operating conditions and the mycotoxin content. Sample were collected from a biogas plant consisting of a three steps production taking place in a bioreactor, post-reactor and a storage tank. Total DNA was extracted from samples belonging to each steps of biogas production. Metagenomic analysis was carried out by analyzing the V4 variable region of bacterial and archaeal 16S rRNA gene. Mycotoxin content was analyzed in maize silage feeding the biogas plant and in the digestate from bioreactor, post-reactor and storage tank by immunoaffinity column clean-up (Myco6in1+®) and detected with liquid chromatography-tandem mass spectrometry (LC-MS/MS). Over 3million high quality reads (about 1Gb) were generated on the Ion Torrent S5 Sequencing System. About 2.4 million reads were assigned for 16S analysis. In detail, metagenomic analysis revealed that Bacteria superkingdom was dominant (~96%) along the production steps, whereas Archaea were less represented (~4%). Within Bacteria the most abundant phylum was Firmicutes, mostly represented by Clostridia, followed by Bacteroidetes and Synergistetes. Within the superkingdom of Archaea, only microorganisms belonging to the phylum of Euryarchaeota were detected. Within Euryarchaeota the dominant genera were Methanosarcina and Methanoculleus. Chemical analysis on maize silage feeding the plants showed an initial mycotoxin contamination by DON (410 µg/kg), FB1 (3570 µg/kg), FB2 (810 µg/kg) and T-2 toxin (20 µg/kg), while AfB1, HT-2 Toxin, NIV, OTA and ZEA were not detected. After the first step of biogas production, a complete reduction of DON and T-2 content was achieved. These preliminary results suggest a possible absorption/degradation of mycotoxins in bioreactor tank and therefore further studies are needed to better elucidate the possible involvement of specific microbial taxa capable of mycotoxins reduction and the enzymatic pathways potentially involved in mycotoxin degradation.

Energy crisis and environmental pollution have led to an increasing interest in renewable energies. Biogas production from plant material, agricultural residual products and food wastes represents one of the most economically attractive alternative technology for biofuel production. Complex consortia of microorganisms are responsible for biomass degradation and biogas production involving several stages. Next-generation high-throughput sequencing provides a powerful tool for dissecting microbial community structure and methane-producing pathways in anaerobic digestion. A taxonomic and functional metagenomic analysis of microbial community residing in an industrial-scale biogas fermenter has been carried out at different steps of biogas production. Sample were collected from an industrial-scale mesophilic plant consisting of a three steps production taking place in a bioreactor, post-reactor and a storage tank. Total DNA was extracted from samples belonging to each steps of biogas production. Metagenomic analysis was carried out by using 16S and shotgun sequencing approach. The 16S datasets were generated by sequencing the bacterial and archaeal V4 hypervariable region. Reads from 16S sequencing were aligned against SILVA ribosomal RNA sequence database by using MALT(1), while shotgun reads were aligned against NCBI-nr protein database by using DIAMOND(2). Taxonomic binning and functional annotation were performed with MEGAN 6 software(3). About 2.9 and 11.5 million high quality reads were generated on the Ion Torrent S5 Sequencing System for 16S and shotgun approach, respectively. Metagenomic analysis revealed that the overall taxa distribution resulting from both sequencing strategies was conserved. In details, the superkingdom of Bacteria was dominant (~93%) along the production steps, whereas Archaea were less represented (~4%). Within the superkingdom of Archaea, only microorganisms belonging to the phylum of Euryarchaeota were detected. Within the key microorganisms involved in methanogenesis, data showed that during biogas production steps the abundance of Methanosarcina genus decreased from bioreactor to storage tank, with a simultaneous increase of Methanoculleus genus. Considering the key methanogenesis pathways, functional analysis supported a shift from acetotrophic methanogens to hydrogenotrophic methanogens. Results showed that the combination of both 16S and shotgun sequencing approach successfully addressed the taxonomical and functional analysis of microbial community, revealing new insights in microbial and functional dynamics during biogas production steps.

Worldwide mycotoxins contamination has a significant impact on animal and human health, and leads to economic losses accounted for billions of dollars annually. Since the application of pre- and post- harvest strategies, including chemical or physical removal, are not sufficiently effective, biological transformation is considered the most promising yet challenging approach to reduce mycotoxins accumulation. Although several microorganisms were reported to degrade mycotoxins, only a few enzymes have been identified, purified and characterized for this activity. This review focuses on the biotransformation of mycotoxins performed with purified enzymes isolated from bacteria, fungi and plants, whose activity was validated in in vitro and in vivo assays, including patented ones and commercial preparations. Furthermore, we will present some applications for detoxifying enzymes in food, feed, biogas and biofuel industries, describing their limitation and potentialities.

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 ?g/kg. OTA was measured in 4 samples of currants (mean value of 6.6 ?g/kg) and 13 samples of raisins (mean value of 1.4 ?g/kg). In one sample of currants and one of raisins from Turkey OTA exceeded the limits set by European Commission of 10 ?g/kg, being contaminated with 12.61 and 15.99 ?g/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.

Fusarium verticillioides and Fusarium proliferatum are the main source of fumonisins, a group of mycotoxins that can contaminate maize-based food and feed and cause diseases in humans and animals. We analyzed the effect of temperature (15°C-35°C), water activity (aw: 0.999-0.93), salinity (0-125 g/l NaCl), pH (5-8) and light of different wavelength (650-390 nm) on the growth, the production of fumonisins B (FB) and the expression of FUM1 and FUM21. For F. verticillioides the highest growth rate was measured at 25°C, aw of 0.998-0.99, 0-25 g/l of NaCl and white light. Optimal conditions for fumonisin production were 30°C, aw of 0.99, 25 g/l of NaCl, pH 5, red and blue light. For Fusarium proliferatum the highest growth rate was measured at 25°C, aw of 0.99 and 0-25 g/l of NaCl, pH 7 and green light. Optimal conditions for fumonisin production were 25°C, aw of 0.998, 0 g/l of NaCl, pH 6, red and blue light. Fusarium verticillioides showed a better adaptability compared to F. proliferatum and was able to produce moderate levels of fumonisins under a wide range of conditions. FUM gene expression not always mirrored FB production, indicating a post-transcriptional mechanism regulationg fumonisin production. The study of the effect of different environmental conditions on toxin production should provide information that can be used to develop strategies to minimize the risk.

Fusarium verticillioides and Fusarium proliferatum are the mainsource of fumonisins, a group of mycotoxins that can contaminate maize-based food and feed and cause diseases in humans and animals.We analyzed the effect of temperature (15-35°C), water activity(aw: 0.999-0.93), salinity (0-125 g/l NaCl), pH (5-8) and light ofdifferent wavelength (650-390 nm) on the growth, the productionof fumonisins B (FB) and the expression of FUM1 and FUM21. ForF. verticillioides the highest growth rate was measured at 25°C, awof 0.998-0.99, 0-25 g/l of NaCl and white light. Optimal conditionsfor fumonisin production were 30°C, aw of 0.99, 25 g/l of NaCl,pH 5, red and blue light. For Fusarium proliferatum the highestgrowth rate was measured at 25°C, aw of 0.99 and 0-25 g/l of NaCl,pH 7 and green light. Optimal conditions for fumonisin productionwere 25°C, aw of 0.998, 0 g/l of NaCl, pH 6, red and bluelight. F. verticillioides showed a better adaptability compared toF. proliferatum and was able to produce moderate levels of fumonisinsunder a wide range of conditions. FUM gene expression notalways mirrored FB production, indicating a post-transcriptionalmechanism regulationg fumonisin production. The study of theeffect of different environmental conditions on toxin productionshould provide information that can be used to develop strategiesto minimize the risk.

Fusarium verticillioides is the main source of fumonisins, a group of mycotoxins that can contaminate maize-based food and feed and cause diseases in humans and animals. The study of the effect of different environmental conditions on toxin production should provide information that can be used to develop strategies to minimize the risk. This study analysed the effect of temperature (15°C-35°C), water activity (a (w): 0.999-0.93), salinity (0-125 g l(-1) NaCl) and pH (5-8) on the growth and production of fumonisins B(1) (FB1), B(2) (FB2) and B(3) (FB3) and the expression of FUM1 and FUM21 in F. verticillioides. The highest growth rate was measured at 25°C, a (w) of 0.998-0.99 and 0-25 g l(-1) of NaCl. Optimal conditions for fumonisin production were 30°C, a (w) of 0.99, 25 g l(-1) of NaCl and pH 5; nevertheless, the strain showed a good adaptability and was able to produce moderate levels of fumonisins under a wide range of conditions. Gene expression mirrored fumonisin production profile under all conditions with the exception of temperature: FUM1 and FUM21 expression was highest at 15°C, while maximum fumonisin production was at 30°C. These data indicate that a post-transcriptional regulation mechanism could account for the different optimal temperatures for FUM gene expression and fumonisin production.

L'aflatossina M1 (AFM1) è il principale metabolita derivante dall'idrossilazionedell'aflatossina B1 (AFB1) presente nel latte di animali alimentati con mangimi contaminati da AFB1 ed è classificato nel gruppo 2B, potenzialmente cancerogeno per l'uomo, dall'AgenziaInternazionale per la Ricerca sul Cancro (IARC). Il livello limite della AFM1 nel latte crudo,trattato termicamente e destinato alla produzione di prodotti a base di latte è fissato a50ng/kg dal regolamento europeo numero 1881 del 2006. Essendo resistente ai comunitrattamenti dell'industria alimentare, la presenza di AFM1 è documentata in tutti i prodottidella filiera lattiero casearia, inclusi yogurt e formaggi, e rappresenta un serio pericolo perla salute. Lo sviluppo di metodi per la riduzione della contaminazione di aflatossine è untema cruciale e attuale, ed è complicato dalla necessità di preservare le qualità organolettichee nutrizionali della matrice trattata. In questo lavoro è stata valutata la capacità degradativadi una laccasi da Pleurotus eryngii verso l'AFM1, sia in buffer che in latte, ed il suo effettosulla componente proteica di questa matrice al fine di verificarne l'applicazione per ilmiglioramento della sicurezza di prodotti lattiero caseari. La riduzione di AFM1 in bufferdi sodio acetato pH 6.5 1mM, a 25°C, è di ca 50% dopo 1h di incubazione e risulta completadopo 72h. Simili risultati sono stati ottenuti in latte, sebbene la cinetica di degradazioneabbia registrato un rallentamento nelle prime tre ore di trattamento. L'analisi dei patternproteici in SDS-PAGE ha evidenziato una riduzione nell'intensità delle bande di ? e ?caseine, di ?-lattoglobulina e sieroalbumina bovina, contemporaneamente alla comparsadi aggregati proteici di peso molecolare superiore ai 200kDa. I dati presentati dimostranoil potenziale applicativo della laccasi per lo sviluppo di metodologie green di degradazionedi AFM1 in prodotti a base di latte e per applicazioni tecnologiche volte al miglioramentodella reologia e alla riduzione della componente allergenica in prodotti lattierocaseari.Parole chiave: sicurezza, latte, laccasi, aflatossina M1, cross-link di proteine, reologia dellatte, allergeni

Fusarium genus is able to produce several metabolites including the emerging mycotoxins beauvericin (BEA) and enniatins (ENs). Due to their ionophoric property, BEA and ENs exert many biological properties, including antimicrobial, insecticidal, and cytotoxic activity in human cell lines, so that they are recently proposed as novel anticancer drugs. BEA and ENs are cyclic hexadepsipeptides with an alternating sequence of three N-metyl-L-amino acids and three D-?-hydroxyisovaleric acids; in BEA the amino acid residues are aromatic N-metyl-pheylalanines, whereas in ENs the amino acid residues are aliphatic N-metyl-valine, -leucine or -isoleucine. Both mycotoxins are synthesized by the multifunctional enzyme enniatin synthetase (ESYN1) representing hybrid system of peptide synthetase and S-adenosyl-L-methionine-dependent N-methyltransferase. Several Fusarium species have been reported to produce ENs, BEA or both, and our hypothesis is that the different production profile depends on the esyn1 sequence. Our aim was to investigate this relation by mean of a bioinformatics approach, based on structural investigations.The esyn1 sequences of 18 Fusarium isolates belonging to 8 different species were extracted from published and unpublished genomes by BLASTN search using as query the available sequences of Fusarium scirpi and Fusarium proliferatum. The protein sequences were predicted using the Sequence Translation tools of the EMBOSS Programs (EMBL-EBI), manually curated using exon/intron boundary predictions from SpliceView (http://bioinfo4.itb.cnr.it/), and confirmed by sequencing the RT-PCR products.The selected sequences have been processed by the web server RaptorX to obtain structural predictions by homology modelling and threading methods. While the full sequences have been submitted for prediction, it has been accomplished only for part of the whole enzyme. In particular, the region containing the N-methyltransferase activity has been not completely structured. A comprehensive analysis of the predicted structures has allowed identifying common structural domains, which have been compared by considering their dihedral backbone angles and by using the tool T-PAD, developed to characterize the protein flexibility and identify hot spot residues responsible for hinge motions. The residue-by-residue flexibility profiles have been analysed by multivariate analysis, to produce a classification by Principal Component Analysis, followed by hierarchical clustering.We have thus identified the conserved structural regions and the pivotal residues responsible for the structural variability. The common domains for each of the 18 esyn1 sequences have been grouped in clusters, and the residues responsible for the classification have been singled out. Overall, we have achieved a comprehensive view of the structural features of the analysed esyn1 sequences, where the structural variability has been related to the sequence variability, and interpreted in terms

Ochratoxin A (OTA) is a nephrotoxic and potentially carcinogenic mycotoxin produced by several species of Aspergillus and Penicillium, contaminating grapes, wine and a variety of food products.We recently isolated from OTA contaminated soil vineyard a novel free-livingstrain of Acinetobacter sp. neg1, ITEM 17016, able to degrade OTA into the nontoxic catabolic product OTalpha (OT?). Biochemical studies suggested that the degradation reaction proceeds via peptide bond hydrolysis with phenylalanine (Phe) release. In order to identify genes responsible for OTA degradationwe performed a differential gene expression analysis of ITEM 17016 grown in the presence or absence of the toxin. Among the differentially expressed genes, 6 peptidases up-regulated at 6 hours were identified. The degrading activity of the carboxypeptidase PJ_1540 was confirmed in vitro in a heterologous system. The enrichment analysis for Gene Ontology terms confirmed that OTA degradation proceeds through peptidase activities and revealed the over-representation of pathways related to Phe catabolism. These results indicate that Phe may represent an energy source for this Acinetobacter sp. neg1strain and that OTA degrading reaction triggers the modulation of further catabolic activities.

The aim of this study was to analyse the transcriptional regulation of enniatins (ENs) production in Fusarium avenaceum. Methods and Results: We develop a new method to quantify ENs in FDM agar medium. We performed an LC/MS/MS analysis to evaluate enniatin A, A1, B, B1 and B4 production by seven F. avenaceum strains and, in a time-course experiment, by ITEM 3404 to analyse the transcriptional regulation of the esyn1 gene. The expression profile, achieved by Real time reverse transcriptase assay, showed an activation of gene transcription at the seventh day of incubation, corresponding to the higher increase of total ENs production. Enniatin B was the most abundant ENs analogues, representing the 90% of total ENs. The relative percentage of ENs remained unaltered during the experiment. Conclusions: We reported a transcriptional regulation of esyn1 responsible for the modulation of ENs biosynthesis. Significance and Impact of the Study: Enniatins are cyclic depsipeptides metabolites with a wide range of biological activities. They are also widespread contaminants in grains and cereals due to infection by enniatin-producing Fusarium species. This is the first article describing the transcriptional regulation of esyn1 gene that modulates ENs production in Fusarium avenaceum and provides new knowledge about the molecular mechanism underlying the biosynthesis of these important fungal metabolites in this toxigenic fungal species. © 2013 The Society for Applied Microbiology.