Ochratoxin A (OTA) is a potent pentaketide nephrotoxin diffusely distributed in food and feed products; it is also carcinogenic, neurotoxic, teratogenic and immunotoxic. The mycotoxin is produced by species of genus Aspergillus and Penicillium. OTA is the primarily mycotoxin risk in wine and dried wine fruits, and the main source of OTA contamination in grapes is A. carbonarius, followed by A. niger and A. welwitschiae. Geographical regions, climatic conditions, crop and pest management, and grape genotypes influence the contamination risk. So, the availability of validate predictive models could be very useful in supporting the optimization of grape management for the mitigation of OTA content in grapes. So far, the only predictive model for OTA risk in grapes is that developed by Battilani et al. [1]. It uses hourly data on air temperature, relative humidity and rainfall as inputs and provides a risk assessment during the growing season. The model has not yet been validated with real field data. In this framework, the OTA model was implemented in a digital platform, developed within the project "Digital Grape", aiming at supporting the management of main agronomic and phytosanitary practices for precision viticulture (https://digitalgrape.it/). The model was elaborated and preliminary tested with experimental data, and successively used to predict OTA risk on 43 Apulian vineyards in two consecutive years (2021-2022). Grape samples were collected at harvest from each field and analyzed for OTA content and A. carbonarius contamination. The results were compared with the Toxin Index generated by the OTA model at harvest time. Chemical analysis evidenced for both years an OTA content always below the legal limit of 2 µg/Kg [2] and A. carbonarius contamination ranging from 0 to 1,1 x 107 CFU/g must, while the Toxin Index varied from 75 to 6097. In general, the correlation between the Toxin Index and the OTA level was low, mainly for the vineyards located in Salento (South of Apulia), while a higher correlation was observed for vineyards in North of Apulia, especially in 2022. On the opposite, a good correlation between the OTA level and A. carbonarius contamination was observed in most of the vineyards in both years. These preliminary results highlighted that the OTA predictive model need to be improved through fine-tuning on experimental data for being useful in managing OTA risk in a smart agriculture system. References 1.Battilani, Paola, and Marco Camardo Leggieri. "OTA-grapes: a mechanistic model to predict ochratoxin A risk in grapes, a step beyond the systems approach." Toxins 7.8 (2015): 3012-3029. 2.Commission Regulation (EC) No 1881/2006 of 19 December 2006 setting maximum levels for certain contaminants in foodstuffs. Aknowledgements This work was partially funded by Puglia Region - Project "Digital Grape - New Digital Technologies and Decision Support Systems for the Improvement of Quality and Sustainability in Viticulture" - P.S.R. Puglia 2014/2020 - Misura 16 - Cooperazione - Sottomisura 16.2 "Sostegno a progetti pilota e allo sviluppo di nuovi prodotti, pratiche, processi e tecnologie".

In recent years, the importance of the conservation of microbial biodiversity has achieved high awareness. Conservation biologists have highlighted that weaknesses in microbial preservation can lead to the loss of valuable bio-resources and gene pools that provide high levels of resistance against changing environmental conditions and adaptation to novel production processes. Community, habitat and intact microbial communites preservation are promising solutions to mitigate and counteract these emerging problems, favouring the biotechnological valorisation of microbiomes. Generally, microbiologists focus their consideration only on culturable microbes and on pure cultures, thus preserving only a tiny fraction of microbial diversity and functional potential of that microbiota. Besides, very few studies have been conducted to study the optimal conditions of storage of microbial consortia and their effect on microbial community structure and the related functional potential. An efficient storing method must be able to maintain microbial strains in a viable state to protect their morphological and genetic stability for a long time under laboratory conditions. Several methods are used for the preservation and maintenance of microorganisms. Usually, lyophilisation and cryopreservation are versatile and widely applicable for a large number of microorganisms. Moreover, the presence of cryoprotectants often increases the survival of microorganisms, and in particular, glycerol and DMSO were shown to protect microbial cells against freezing damage. This study aimed to evaluate the effectiveness of freezing procedures to cryopreserve the microbiota isolated from the surface of an Apulian cured sausage, using glycerol or DMSO as cryoprotectants and two different storing temperatures (-80 °C and -135 °C) for the short and mid-term period. The microbial population of the sausage was studied by culture-dependent method and metagenomic analysis comparing analyses before and after storing period. Results indicated that after 1 and 8 months of cryopreservation, the viability of the bacterial population of microbial consortium decreased at the same rate for both cryoprotectants and storing temperatures. A similar trend was also observed for fungi and yeasts with the exception of storage conditions that include glycerol and temperature of -135°C, which better preserve the viability of yeasts and fungi after eight months. This study pointed out the first scientific evidence on the effect of cryopreservation of the whole microbial consortia collected from the surface of cured sausages. Part of this work was granted by the European Commission - NextGenerationEU, Project "Strengthening the MIRRI Italian Research Infrastructure for Sustainable Bioscience and Bioeconomy", code n.IR0000005.

Fermented foods and beverages represent an important part of the gastronomic heritage, about one-third of human diet, but also a dynamic and constantly evolving sector, offering solutions to improve the well-being and nutrition of different population targets. Fermentation also makes it possible by promoting sustainable innovation and enhancing the quality of food and beverages. This family of microbial-based bioprocesses combine traditional knowledge, food microbiology and food sciences, also concretising interesting models of study for ecology, biodiversity and biological sciences. PIMENTO COST Action CA20128 is a pan-European transdisciplinary community that works to i) study the diversity of European fermented products, ii) create an original synthesis in the definition of the benefits and risks associated with these product categories, and iii) analyse the dynamics of innovation in the field. Here, we report the recent advances of the PIMENTO project, underlying how the selected activities from a constellation of Italian National Recovery and Resilience Plan (NRRP) projects financed by the European Commission's Next Generation EU programme contribute to shape new trajectories in research activities in the field, particularly about: a) food microbiomes preservation (SUS-MIRRI.IT); b) fermented products in marginal areas (AGRITECH); c) fermentation for sustainable food systems (ONFOODS); d) sensors to promote and speed up innovation in the sector (iNEST).

Eukaryotic microorganisms of protechnological interest in food chains represent essential resources for manufacturing fermented foods and beverages, helping to determine the quality and safety of finished products. In the first part of this contribution, an examination of the role of yeasts and filamentous fungi in the various categories of traditional fermented products is proposed, with a particular interest in geographical indications and typical European products. The overview looks at the contribution with respect to the main components of quality: sensorial, nutritional, functional and hygienic-sanitary. The relevance of microbiomes of fermentative interest containing eukaryotic microbes is then discussed, with particular reference to the ex situ preservation practices of microbiomes associated with specific productions, also in the framework of the activities of microbial collections. In fact, the interest in conserving microbiomes to preserve microbiodiversity and their functionality, as also for study their activities and to guarantee adequate biotechnological valorisation, is increasingly evident. This is an important future challenge involving numerous biological resource centers worldwide. Finally, with a series of case studies, the importance of microbiological cross-over is underlined as a driver of innovation in the sector of fermented products, as a strategy for the development of new products and as an element of regionalisation of non-traditional productions. In other words, it is underlined how the preservation, characterisation and enhancement of microbial diversity can represent a reservoir of solutions capable of improving the attributes of existing products, increasing the added value and promoting the design of new foods and fermented beverages. "Gulden Goksen would like to acknowledge the direct support by COST (European Cooperation in Science and Technology; www.cost.eu), in the framework of COST Action CA20128 (Promoting Innovation of ferMENTed fOods; https://fermentedfoods.eu/)."

Sequencing fungal genomes has now become very common and the list of genomes in this manuscript reflects this. Particularly relevant is that the first announcement is a re-identification of Penicillium genomes available on NCBI. The fact that more than 100 of these genomes have been deposited without the correct species names speak volumes to the fact that we must continue training fungal taxonomists and the importance of the International Mycological Association (after which this journal is named). When we started the genome series in 2013, one of the essential aspects was the need to have a phylogenetic tree as part of the manuscript. This came about as the result of a discussion with colleagues in NCBI who were trying to deal with the very many incorrectly identified bacterial genomes (at the time) which had been submitted to NCBI. We are now in the same position with fungal genomes. Sequencing a fungal genome is all too easy but providing a correct species name and ensuring that the fungus has in fact been correctly identified seems to be more difficult. We know that there are thousands of fungi which have not yet been described. The availability of sequence data has made identification of fungi easier but also serves to highlight the need to have a fungal taxonomist in the project to make sure that mistakes are not made.

Motivation. Although the knowledge of the contribution of molds in fermented foods is still very limited, compared to bacteria and yeasts, several fungal species are commonly used which contribute to the improvement of food safety, nutritional value, organoleptic quality, and food preservation. However, filamentous fungi can also be considered food contaminants leading to food spoilage and the production of toxic metabolites. Penicillium is among the predominant fungal genera capable of growing on the surface of dry fermented meats, and several species, such as P. nalgiovense, have been domesticated by continuos selection for beneficial traits. Here we investigate the genome of six Penicillium species isolated from Italian salami, which have been shown to be adapted to the curing conditions occurring during the production of fermented sausage, with the aim of characterizing the differences in secretomic potential, which may have been related to the maturation of meat products. Methods. The genome sequencing and annotation of Penicillium brevicompactum ITEM 18316, P. biforme ITEM 15300, P. chrysogenum ITEM 4680, P. cvjetkovicii ITEM 18317, P. nalgiovense ITEM 18323, P. salamii ITEM 15291, and P. solitum ITEM 18327 isolated from dry cured meat was part of a collaborative sequencing project involving JGI, PNNL and CNR-ISPA. Draft genomes and transcriptomes were generated using Illumina technology. Genome assemblies were masked for repeats using RepeatMasker, with the RepBase library and RepeatScout, and the measures of genome assembly completeness were performed with BUSCO. Nuclear genomes were annotated using the JGI Annotation pipeline, and further improved. Protein-coding gene models were generated using a combination of ab initio, homology-based, and transcriptome-based gene predictors. Predicted proteins were functionally annotated using SignalP, TMHMM for transmembrane domains, InterProScan for protein domains, and Blastp alignments against the NCBI NR, SwissProt, KEGG, and KOG. Transcription factors were assigned based on Pfam. GO terms were assigned based on InterPro and SwissProt hits. TCDB and MEROPS were used for transporter and peptidase classifications, respectively. CAZymes, secondary metabolites, and cytochrome 450 subfamilies were also annotated and antiSMASH fungi was used to improve cluster detection. Subcellular localization of proteins was predicted with DeepLoc and proteins with extracellular localization were realigned with Diamond and reannotated with eggNOG-mapper. Proteins and peptide domains with interesting catalytic or antimicrobial activities that can describe the contribution of molds in meat curing have been interrogated and retrieved. Additioanlly, the mycotoxin production potential of each fungal species was also evaluated. Orthofinder analysis was also implemented to provide comprehensive statistics for the comparative genomics of the six Penicillium isolates, and genome of other isolates of the same species deposited on NCBI, if available. Results. The draft genome assembly resulted in 165 to 750 contigs and 156 to 692 scaffolds for Penicillium brevicompactum ITEM 18316 and P. salamii ITEM 15291, respectively. The gene prediction identified 9657 to 12781 protein-coding genes for P. cvjetkovicii ITEM 18317 and P. biforme ITEM 15300, respectively. Using whole genome sequence information and bioinformatics tools, various genomic features were explored to characterize the secretory potential of the studied fungal species, focusing on several interesting properties. Complete clusters for mycotoxin biosynthesis were identified in P. brevicompactum ITEM 18316 (mycophenolic acid), and P. biforme ITEM 15300 (PR-toxin). Protease and peptidase enzymes that complete the hydrolysis of proteins and peptides into amino acids, and lipase and esterase enzymes that hydrolyze fats to release the fatty acids have been catalogued. The catalytic activities of catalase, amino-oxidase and thioredoxin, which play important roles in the modulation of oxidation of lipids, biogenic amines, and allergenic proteins, respectively, have been listed. P. brevicompactum ITEM 18316, P. cvjetkovicii ITEM 18317 and P. salamii ITEM 15291, excreted a gamma glutamyl transpeptidase/transferase enzyme that may contribute to glutamate formation that is well known for improve sensory characteristic of cured meat. Peptides or domains related to antifungal proteins, killer toxins, and bubble proteins, which may contribute to the competitive advantage of fungal growth on the meat matrix over other microorganisms, were investigated. P. biforme ITEM 15300 and P. nalgiovense ITEM 18323 secreted the same PAF antifungal proteins as P. chrysogenum, meanwhile, P. salamii ITEM 15291 appears to excrete an antifungal protein, partially similar to PaAFPa of P. expansum and PeAFPb of P. digitatum. The functionally diverse activities secreted by these six Penicillium isolates, will shed lights on the role they may play in the dry-cured meat environment.

Ochratoxin A (OTA) contamination and the associated issues of food security, food safety and economic loss are widespread throughout the world. The occurrence of OTA depends on ochratoxigenic fungi, foodstuffs and their environment. In this review, natural occurrence and control strategy of OTA, with a focus on the impact of environmental factors, are summarized. First, this manuscript introduces potentially contaminated foodstuffs, including the emerging ones which are not regulated in international legislation. Secondly, it gives an update of native producers based on foodstuffs and OTA biosynthesis. Thirdly, complicated environmental regulation is disassembled into individual factors in order to clarify their regulatory effect and mechanism. Finally, to emphasize control OTA at all stages of foodstuffs from farm to table, strategies used at crop planting, harvest, storage and processing stages are discussed.

Filamentous fungi possess a wide diversity of metabolic pathways, among which the production of mycotoxins and the resultant contamination of agricultural commodities cause severe health impacts on humans and animals. Understanding the biology, ecology and genetics of mycotoxins biosynthesis is fundamental to counteract their spread in food and feed products and reduce the human and animal health risk. The gene clusters responsible for the biosynthesis of mycotoxins of agricultural importance, including aflatoxins, fumonisins, ochratoxins, patulin, citrinin and trichothecenes, have been mostly identified and characterized. However, due to the complex organization of fungal secondary metabolisms and interaction with climatic, environmental and biotic factors, numerous new researches have been recently published on structural, regulatory and epigenetics mechanisms underlying mycotoxin biosynthesis. This review provides an overview of the recent new insight into understanding genes, molecular mechanisms and factors involved in biosynthesis regulation of the principal mycotoxins.

In recent years, the cultivation of tropical fruit crops has increased in the Mediterranean basin, especially in southern Italy. In surveys conducted from 2014 to 2019 woody canker and shoot blight were observed on mango plants (cvs. Kent, Keitt, Sensation, Osteen, and Kensington Pride) and litchi plants (cvs. Way Chee and Kwai Mai Pink) cultivated in Sicily. Botryosphaeriaceae and Diaporthaceae were consistently isolated from symptomatic samples. Morphological characterization and multi-locus phylogenies using three genomic loci (a portion of translation elongation factor 1-? gene, a portion of the ?-tubulin gene, and an internal transcribed spacer) identified these fungi as Neofusicoccum parvum, Lasiodiplodia theobromae, Botryosphaeria dothidea, Diaporthe foeniculina, and Diaporthe baccae on mango and Diaporthe foeniculina and Diaporthe rudis on litchi. Pathogenicity tests on healthy mango (cv. Kensington Pride) and litchi (cv. Way Chee) plants demonstrated the pathogenicity of the isolates used in the study, and Koch's postulates were fulfilled for all pathogens. To our knowledge, this is the first report of L. theobromae, B. dothidea, and Diaporthe species on mango in Italy and the first report worldwide of woody canker and shoot blight caused by D. foeniculina and D. rudis on litchi plants.

In this study, eight lactic acid bacteria (LAB) strains, previously isolated from traditional and gluten-free sourdoughs, and selected for their potential in improving the sensory and rheological quality of bakery products, were screened against some common spoilage agents. The anti-mould activity was tested using strains of the species Fusarium graminearum, Aspergillus flavus, Penicillium paneum and Aspergillus niger. Regarding the antibacterial activity, it was assessed against four strains of the species Escherichia coli, Campylobacter jejuni, Salmonella typhimurium and Listeria monocytogenes. Furthermore, LAB strains were evaluated for their ability to produce exopolysaccharides, which are gaining considerable attention for their functional properties and applicability in different food industrial applications. A strain-specific behaviour against the moulds was observed. In particular, F. graminearum ITEM 5356 was completely inhibited by all the LAB strains. Regarding the antibacterial activity, the strains Leuconostoc citreum UMCC 3011, Lactiplantibacillus plantarum UMCC 2996, and Pediococcus pentosaceus UMCC 3010 showed wide activity against the tested pathogens. Moreover, all the LAB strains were able to produce exopolysaccharides, which were preliminarily characterized. The assessed features of the LAB strains allow us to consider them as promising candidates for single or multiple starter cultures for food fermentation processes.

Relazione tecnica delle attività espletate durante il primo anno del progetto con primi risultati ottenuti nei diversi workpackage e task del progetto.

Ochratoxin A (OTA) is a well-known mycotoxin with wide distribution in food and feed. Fungal genome sequencing has great utility for identifying secondary metabolites gene clusters for known and novel compounds. A comparative analysis of the OTA-biosynthetic cluster in A. steynii, A. westerdijkiae, A. niger, A. carbonarius, and P. nordicum has revealed a high synteny in OTA cluster organization in five structural genes (otaA, otaB, ota, otaR1, and otaD). Moreover, a recent detailed comparative genome analysis of Aspergilli OTA producers led to the identification of a cyclase gene, otaY, located in the OTA cluster between the otaA and otaB genes, encoding for a predicted protein with high similarity to SnoaLs domain. These proteins have been shown to catalyze ring closure steps in the biosynthesis of polyketide antibiotics produced in Streptomyces. In the present study, we demonstrated an upregulation of the cyclase gene in A. carbonarius under OTA permissive conditions, consistent with the expression trends of the other OTA cluster genes and their role in OTA biosynthesis by complete gene deletion. Our results pointed out the involvement of a cyclase gene in OTA biosynthetic pathway for the first time. They represent a step forward in the understanding of the molecular basis of OTA biosynthesis in A. carbonarius.

The MycoKey-Mycotwin Conference (Horizon 2020 projects) will take place over three days, (9-12 November 2021) with six open scientific sessions dedicated to different aspects/topics aiming to cover the management of mycotoxin risk in food and feed chain in an holistic view. This book is a collection of the best and interesting achievements of the MycoKey project and the dissemination messages on mycotoxins and toxigenic fungi of MycoTwin project. Worldwide and renowned international scientists will present in the following sessions: 1) biodiversity and functional genomic of toxigenic fungi 2) innovative monitoring methods for toxigenic fungi and mycotoxins 3) prevention and control of toxigenic fungi and myctoxins in the field 4) food and feed remediation, intervention and processing for mycotoxin reduction Combined to the Conference is the 2nd International Forum on Mycotoxins in Animal Production that will deal with the main mycotoxins effect on animal production at global level.

The widespread use of Next-Generation Sequencing (NGS) for fungal genome sequencing has led to identification of SM clusters for known metabolites as well as a significant number of novel predicted SM gene clusters. Fungal genome sequencing has great utility for identification of secondary metabolites gene clusters for known as well as novel compounds. Ochratoxin A (OTA) is a well-known mycotoxin with wide distribution on food and feed. A comparative analysis of gene cluster structure in 19 Aspergillus and 2 Penicillium OTA producers has revealed a high synteny across these species in OTA cluster, encoding five structural genes: otaA, otaB, otaC, otaR1, and otaD. Furthermore, we identify a previously undescribed additional gene. The new otaY gene is located between the otaA and otaB genes and encodes a protein with predicted SnoaL domain. These snoaL-domain containing proteins have been shown to catalyze ring closure steps in the biosynthesis of polyketide antibiotics produced in Streptomyces. Gene expression analysis has demonstrated an upregulation of the cyclase gene in A. carbonarius under OTA permissive conditions, consistent with the expression trends of the other OTA cluster genes. The role of in OTA biosynthesis has been demonstrated by CRISPR/Cas9 complete gene deletion. The presented results reveal for the first time the involvement of a cyclase gene in OTA biosynthetic pathway and redefine the structure of the OTA core cluster, consisting in six genes.

Prediction and warnings are important for growers to timely respond to treats and optimize the growth of crops. The MycoKey app was developed to aid mycotoxin risk mitigation by growers, grain collectors, governmental planners and policy makers. The MycoKey app predicts the amounts of the most important mycotoxins in winter wheat (DON) and grain maize (Aflatoxin B1 and Fumonisin) based on local weather data and predictive models. It provides direct links to the scientific articles describing the models in detail. Both access to the platform and the MycoKey app is free of charge and user data are private. Enlist at https://akkerweb.eu/en-gb/ and download the free MycoKey app. Growers can calculate the predicted amount of mycotoxins in their crops with local weather information and relevant agronomic measures such as ploughing or the level of plant resistance based on a list of cultivars. Grain collectors, governmental planners and policy makers have access to public databases including satellite data, and can calculate mycotoxin risks based on weather data and land use (if publicly available). Recalculation allows integration of management strategies in the risk model and calculations of "what if" scenarios. The risk analysis was performed for the mycotoxin DON in winter wheat and in 2019 and 2020 DON mycotoxin risks for winter wheat were low in the Belgium and the Netherlands. In contrast in 2021 conditions were more conducive for Fusarium and the risks for DON were much higher particular in the northern part of the Netherlands. The Mycokey app was used to create awareness and helped to initiate responses in the chain to mitigate mycotoxin risks in the Netherlands. The early warning was made possible by filling the gap between forecast and harvest time using historic weather available for the last 40 years. These historic data can also be applied to assess the impact of weather in climate change scenarios. The need for an integrated approach to mitigate mycotoxin risks in crop management systems will be discussed.

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.

Recently, Aspergillus flavus arise as an emerging problem for maize in Southern Europe. After the first European aflatoxins outbreak in 2003, followed by severe contamination in 2012 and 2015, the prevention of aflatoxins in maize field became a relevant issue. Competitive exclusion of toxigenic Aspergillus flavus by atoxigenic strains is worldwide accepted as one of the most effective action to minimize aflatoxins risk. USA, Africa and Italy have developed some commercial biocontrol agent products and Eastern Europe countries start working in the same direction. This study aimed to select an A. flavus biocontrol agents in Romania. Therefore, Aspergillus section Flavi strains were isolated, identified and characterized, from 139 maize flour samples, representative of the whole Romanian maize growing areas. Deletions of the aflatoxins gene cluster were investigated by multiplex PCR analyses. Microsatellite alleles were bioinformatically analysed to depict the population structure. 169 strains were confirmed as A. flavus; 71 strains seems to lack at least one gene of the aflatoxins cluster. Competition tests, involved 7 chemically confirmed atoxigenic strains with huge deletions in aflatoxins cluster. Microsatellites analysis identify two main group by bayesian clustering, genetic distance-based analysis and population assignment. Strain 36.3 was selected as potential biocontrol agent, among the 7 atoxigenic strains that exhibited high aflatoxins reduction (80-96%) during in vitro competition tests. Thermally treated sorghum coated with the inoculum of this atoxigenic strain was distributed in maize field trials (approximately 1 ha) in Romania (2018 and 2019). Since, very low aflatoxins contamination was measured, even in untreated fields, during both years, the biocontrol efficacy needs to be confirmed by further field data.

Negli ultimi anni, è crescente l'interesse verso la biodiversità microbica, la cui preservazione sta assumendo sempre più maggiore importanza per il miglioramento della qualità e sicurezza degli alimenti, per lo sviluppo di biotecnologie verdi e della bioeconomia agro-alimentare. Infatti, i microrganismi di interesse agro-alimentare svolgono un ruolo chiave in molti aspetti della produzione agraria quali fertilità del suolo, nutrizione delle colture, controllo biologico e fertilizzanti biologici. Inoltre, i microrganismi possono svolgere un ruolo importante anche nella produzione di alimenti trasformati, quali latte, formaggio, vino, olio, pane, carne; ed durante la loro conservazione, in quanto ne possono causare alterazione, possono produrre metaboliti tossici, ed alcuni possono rivelarsi microrganismi patogeni. L'esistenza e la biodiversità dei microrganismi sono funzionali al mantenimento degli organismi viventi sulla terra, ed alla sostenibilità dei diversi cicli biologici. La conservazione delle risorse genetiche microbiche e la loro custodia svolgono, dunque, un ruolo significativo sia per migliorare le conoscenze, sia per sfruttamenti e sviluppi futuri nelle produzioni agrarie ed alimentari. La Collezione ITEM di microorganismi di interesse agro alimentare del CNR-ISPA, comprende più di 13.500 ceppi microbici, di cui 7.600 pubblici. Per queste accessioni pubbliche, le informazioni relative sono consultabili sul database: http://www.ispa.cnr.it/Collection. La Collezione ITEM è stata riconosciuta a livello internazionale nell'ambito della European Culture Collection Organization (ECCO) e della World Federation of Culture Collection (WFCC). Inoltre, dal 2018, è certificata ISO 9001: 2015 (Cert. N 35674/17/S). Tutti i ceppi sono conservati in purezza e in duplicato mediante due sistemi di conservazione: crioconservazione a - 150 ° C e colture fresche a 4 ° C. La biodiversità microbica (funghi, lieviti e batteri) rappresentata in collezione è molto ampia e ricopre i principali sistemi agroalimentari del Mediterraneo. Tra i microrganismi conservati, si ritrovano miceti dei generi Aspergillus, Alternaria, Beauveria, Fusarium, Penicillium, Trichoderma, lieviti dei generi Saccharomyces, Hanseniaspora, batteri dei generi Acetobacter, Lactobacillus, Oenococcus, Pseudomonas, Staphylococcus, tutti di rilevante importanza patologica, tossicologica e agroalimentare. Alcuni di loro sono coinvolti in malattie vegetali e animali, altri sono responsabili della contaminazione da micotossine di colture e prodotti alimentari, nonché della produzione di vari metaboliti secondari con diverse attività biologiche (es. antibiotici, entomo, fito e zoo-tossine). Altri, ed in particolare lieviti e batteri autoctoni (compresi i probiotici), sono stati preservati e caratterizzati per la loro applicazione nei settori dell'industria alimentare e delle bevande (es. alimenti funzionali, vini tipici pugliesi, olive da tavola e latticini). La collezione ITEM garantisce una fonte genetica originale, unica per la salvaguardia della biodiversità microbica, per il miglioramento delle conoscenze sui microrganismi tossigeni, patogeni ed alterativi, e per le possibili applicazioni biotecnologiche. Gli approcci multidisciplinari utilizzati hanno permesso di ottenere di recente una caratterizzazione biologica, molecolare e tossigena di miceti dei generi Fusarium, Aspergillus, Penicillium e Alternaria, e l'identificazione di nuove specie tossigene (1). Peraltro, sono stati ottenuti risultati importanti per microrganismi di interesse agroalimentare e agroindustriale quali: batteri lattici usati come starter nella produzione di pane senza lievito (2) e di vini con aroma migliorato (3); funghi filamentosi utili per la stagionatura delle carni (4); ceppi batterici e lieviti per la fermentazione delle olive, brevettati per sfruttamento industriale (5), così come è stato brevettato un ceppo probiotico afferente al genere Lacticaseibacillus (6), pronto per l'impiego nell'industria alimentare. Bibliografia 1.Moretti & Susca 2017. Mycotoxigenic Fungi Methods and Protocols, 1542 (pp.383). 2.De Bellis et al. 2019. Foods, 8(2), 70 3.Cappello et al. 2017. Int. J. Food Microbiol. 243: 16-27. 4.Magistà et al. 2016. Int. J. Food Microb. 231, 33-41. 5.Bleve et al. (2015). Food Microbiology, 46:368-382. 6.Valerio et al. 2015. J Funct. Foods, 17, 468-475.

Filamentous fungi are able to synthesise a remarkable range of secondary metabolites, which play various key roles in the interaction between fungi and the rest of the biosphere, deter-mining their ecological fitness. Many of them can have a beneficial activity to be exploited, as well as negative impact on human and animal health, as in the case of mycotoxins contaminating large quantities of food, feed, and agricultural products worldwide and posing serious health and eco-nomic risks. The elucidation of the molecular aspects of mycotoxin biosynthesis has been greatly sped up over the past decade due to the advent of next-generation sequencing technologies, which greatly reduced the cost of genome sequencing and related omic analyses. Here, we briefly highlight the recent progress in the use and integration of omic approaches for the study of mycotoxins bio-synthesis. Particular attention has been paid to genomics and transcriptomic approaches for the identification and characterisation of biosynthetic gene clusters of mycotoxins and the understanding of the regulatory pathways activated in response to physiological and environmental factors leading to their production. The latest innovations in genome-editing technology have also pro-vided a more powerful tool for the complete explanation of regulatory and biosynthesis pathways. Finally, we address the crucial issue of the interpretation of the combined omics data on the biology of the mycotoxigenic fungi. They are rapidly expanding and require the development of resources for more efficient integration, as well as the completeness and the availability of intertwined data for the research community.

Le applicazioni 'cross-over' nel campo della microbiologia degli alimenti rappresentano un approccio che prevede l'utilizzo di un microrganismo isolato/impiegato come starter in uno specifico processo di fermentazione tradizionale nel new product development e/o per migliorare la qualità e la sicurezza in un'altra filiera o altra produzione alimentare (Dank et al. 2021). Il settore viti-vinicolo da sempre manifesta un elevato interesse nello studio e valorizzazione della biodiversità microbica, fondamentale per garantire qualità e sicurezza delle produzioni enologiche. A questo riguardo, particolare rilievo, per importanza e versatilità, è riconosciuto a ceppi di lievito Saccharomyces e non-Saccharomyces, oltre che a diverse specie di batteri malolattici (Tempère et al. 2018; Berbegal et al. 2020; Capozzi et al. 2021). Per decenni è stato compiuto uno sforzo continuo in tutto il mondo per preservare e valorizzare la diversità microbica associata ai vini tradizionali e alle Indicazioni Geografiche, con particolare riguardo a studi sulla microflora associata a specifici vitigni autoctoni. Ad oggi, questo potenziale biotecnologico è stato spesso valorizzato per la produzione di vino, ma resta latente per possibili applicazioni in settori più o meno attigui, quali, ad esempio, la produzione di vini speciali, birra, prodotti da forno, fruity wines e bevande fermentate. Viste le potenzialità di questo tipo di approccio, presenta grande importanza l'elaborazione di una rassegna sistematica della letteratura scientifica, tale da evidenziare le tendenze esistenti e le possibili prospettive future. Accanto alla discussione sui risultati disponibili da studi su applicazioni "crossover" già documentati in letteratura (Capozzi et al. 2016; Agarbati et al. 2020; Canonico et al. 2021; Vilela et al. 2020), la presente revisione propone, inoltre, i) una panoramica della diversità microbica di potenziale interesse e delle caratteristiche fenotipiche ad essa associate funzionali alla progettazione di strategie di 'cross-over' ed ii) evidenze scientifiche da progetti in corso di realizzazione (acronimi di progetto 'INVISPUBA', 'SPUMAPULIA' e 'BE^2R, finanziati dalla Regione Puglia attraverso il P.S.R. 2014/2020 -Misura 16.2). Nel complesso, l'analisi qui proposta tende ad evidenziare l'alto potenziale di innovazione associato alla 'microbiodiversità' enologica, sottolineando la possibilità di esplorare percorsi di carattere regionale, che consentano di ottenere produzioni innovative da contesti produttivi tradizionali attraverso lo scambio di risorse microbiche. L'approccio proposto rimarca, inoltre, l'importanza delle collezioni microbiche (De Vero et al. 2019) nella valorizzazione delle risorse microbiche connesse a determinati contesti geografici e il loro contributo per lo sviluppo della bio-economia e di percorsi di innovazione sostenibile.