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In Europe, as well as in many other western countries, jellyfish (JF) are not a traditional food, they are not usually consumed and the JF market is probably limited to Asiatic communities. The use and marketing of JF in Europe is still hindered by the regulation on novel food (Commission Regulation EU 2283/2015) and mainly by the absence of standard methods for the treatment and processing of the raw material according to the EU safety standards. JF is considered a traditional food in Asian countries, such as China, Japan, Korea, Thailand which host the majority of consumers and producers. Although JF did not make its way to European consumers, some JF species originating from the EU were exported to East Asia for consumption. Traditionally JF are soaked in a mixture of NaCl and aluminum salts. This procedure reduces the water content and changes the JF gelatinous tissue into the consistency expected for the final edible product. This product is generally characterized by a crispy and firm texture highly appreciated by the Eastern market. However, aluminium, that is so extensively used in this procedure, is supposed to cause memory impairment and cognitive dysfunctions, which would lead to neurodegenerative diseases such as Alzheimer's and Parkinson's disease. Then, a reduction of dietary intake of aluminium is highly recommended. In Europe a Tolerable Weekly Intake (TWI) was established of 1 mg aluminium /kg body weight/week; however, the limits could even be more restrictive for overexposed populations. Then, new processes have to be designed to obtain final products observing safety and quality standards, maintaining nutritional traits, having sensory properties suitable for Western consumers. This is particularly true for raw material that has no history as food in western Countries and specifically in the European area (Torri et al., 2019). In within the GoJelly H2020 project, a new process and jellyfish-based products were developed.

The massive blooms of jellyfish (JF), that more and more frequently explode in European seas, negatively impact human health and activities in coastal waters, including aquaculture. As an alternative, these marine gelatinous organisms, should be viewed through a more positive perspective as a new important bio-resource. Indeed, known for their nutritional and medical value in the Chinese pharmacopeia, increasing attention has been pointed to jellyfish as an unexploited source of essential nutrients, novel bioactive metabolites, and lead compounds. Within the H2020 European GoJelly project, JF and their biodiversity, were analysed as an available and abundant source of new natural health-promoting compounds. Different species, Rhizostoma pulmo (Macrì, 1778) that undergoes recurrent outbreaks in the Mediterranean coastal waters, and the zooxanthellatae JF species, as Cotylorhiza tuberculate and Cassiopea andromeda, this last one not native of Mediterranean Sea), were considered as source of new bioactive compounds, both proteinaceous (such as collagen) and non-proteinaceous compounds, as the hydroalcoholic-soluble extracts. R. pulmo peptides were analysed for their antioxidant activity in vitro and in HEKa cell cultures, and the potential immunomodulatory activity of low molecular weight peptides were investigated in monocytes-macrophages couture system (De Domenico et al., 2019). Hydroalcoholic extracts from C. tuberculata and C. andromeda moreover, were assayed for their cytotoxicity effects on cancer cells and for their ability to modulate Gap Junction Intercellular Communication (GJIC) (De Rinaldis et al., in preparation).

La caratterizzazione e l'utilizzo a livello industriale di ceppi selezionati di lieviti e batteri lattici, definiti colture starter, è ormai una parte fondamentale delle biotecnologie applicate alle produzioni enologiche. Come starter sono definite le biomasse microbiche che servono a guidare un determinato processo fermentativo. I vantaggi del loro uso in enologia sono molteplici: i) consentono di condurre la fermentazione con maggiore rapidità; ii) controllano il processo evitando lo sviluppo di fermentazioni anomale ad opera di microrganismi indesiderati e/o alteranti; iii) diminuiscono le problematiche relative ai rallentamenti e agli arresti di fermentazione; iv) permettono di ottenere un prodotto finale di caratteristiche organolettiche costanti.

The setting up of the ARGO detector at the YangBaJing Cosmic Ray Laboratory (4300 m a.s.l., Tibet, P.R. China) has been completed during the last spring (2007). It consists of a central carpet made of 130 identical sub-units of 12 RPCs each (a "cluster"), covering a surface of about 5800 m2 with 93% active area, and a guard ring of 24 further clusters of the same type surrounding the central carpet with a lower sampling density. Signals are picked up by external electrodes of small size, thus allowing the sampling of EAS with high space-time granularity. Shower events are detected at a trigger rate of about 4 kHz. Events with a few particles detected by a single cluster are counted in scaler mode on a time base of 500 ms. The intrinsic modularity of the ARGO detector allowed us to collect data even during the setting-up period, using only the central carpet (or even part of it). Some preliminary results from the analysis of events collected in a few months of data taking are presented. © 2008 Elsevier B.V. All rights reserved.

Following the completion of the 5700m 2 RPC carpet-like AS array at YangBaJing this year, a further plan based on the ARGO Carpet is raised. It characterizes with its high altitude site, its full-coverage detector array and the multi-parameter measurements. Apart from studies on TeV and sub-TeV energy range, UHE Gamma-ray sources and the Knee Physics will become its characteristic subjects. In the first phase of the plan, the Yangbajing Super Complex Array (YSCA) will include a 10 4 m 2 RPC Carpet, five indoor μ -detectors (∼170 m 2 each), four big outdoor μ -detectors (∼432 m 2 each) and a traditional field scintillation detector array surrounding the Carpet. Monte-Carlo study shows that, with such an array in YBJ (4300m a.s.l.), γ -ray induced air showers can be separated from proton induced air showers clearly using only the observed electron and μ data event by event. To assist the classification of UHE air showers induced by different primary nuclei groups without severe interference by Composition/Model Entanglement, a second phase plan having a Central Burst Detector Array (CBDA) and some pulse shape detectors is schemed.

Xylella outbreak in Puglia is ravaging olive plants and research is trying different paths moving through serious social and practical difficulties. The laboratory of Botany in Lecce, in collaboration with CNR ISPA local section, University of Firenze and University of Neuchatel supports the scientific community effort investigating the Olea europea xylem lumen through microscopic analysis. This apoplastic space is the channel through which the pathogen may be targeted by drugs but also a battleground on which the plant itself deploys its natural defensive barriers. In particular woody plants are able to selectively block the traffic through vessels with an active process known as tylose. It is known that tylose impairs water transport during the development of Pierce's disease (PD), caused by Xylella fastidiosa in grape. The cavitation-induced embolisms of xylem due to the bacterial colonization appears to develop before tyloses (Perez-Donoso et al 2016), anyhow both embolisms and tyloses develop with the same pattern and correlate to symptoms. Scanning Electron Microscope observations performed in the xylem of olive plants allowed the identification of different forms of xylem defects, from tylose to biotic obstructions and allowed the correct interpretation of vessels obstructions visualized at low magnification by optical microscopy and confocal microscopy. A visual screening of sections from branches of the three most representative Olea europea cultivars present in the infected area of South Puglia, Cellina di Nardò, Leccino and Ogliarola salentina, was performed. The results will be discussed. Essential bibliography Perez-Donoso, A. G., Lenhof, J. J., Pinney, K., Labavitch, J. M. (2016) Vessel embolism and tyloses in early stages of Pierce's disease. AUSTRALIAN JOURNAL OF GRAPE AND WINE RESEARCH Volume: 22 Issue: 1 Pages: 81-86

Table olives have been a component of the Mediterranean diet for centuries and their consumption is increasing worldwide. They contain several nutritional components such as unsaturated fatty acids, vitamin E, triterpenic acids, sterols, polyphenols and carotenoids [1]. The content of these bioactive molecules depends on the olive variety, the ripening stage of the olive fruit, the cultivation conditions and the processing method [2]. There are numerous commercial preparations of table olives and in a recent work we have developed and validated a new procedure for table olive production based on the use of selected autochthonous microbial starters and a productive process optimized to mimic the microbial evolution observed during spontaneous fermentations. [3]. In this work, we characterized some bioactive compounds and coloring pigments of fermented black table olives, belonging to two Italian (Cellina di Nardò and Leccino) and two Greek (Kalamàta and Conservolea) cultivars. Greek table olives resulted richer in carotenoids, vitamin E and triterpenic acids (maslinic and oleanoic acids) than Italian cv, contents ranged from 2.28 to 6.91 µg/g fw (edible portion), 53.64 to 59.15 µg/g fw and 2.19 to 3.28 mg/g respectively. Polyphenol compounds are known to be responsible for the bitter taste of the olive fruit and most of them are involved in table olives colour changes. In all tested black table olive cultivars, polyphenols content ranged from 0.87 mg/g fw (in Conservolea) to 23.49 mg/g fw (Cellina di Nardò). Moreover, we analysed the content of tyrosol, hydroxytyrosol, oleuropein aglycone, verbascoside, isoverbascoside, rutin, pyrocathecol, luteolin glucoside, quercetin, cyanidine rutinoside and cyniadine glucoside. In particular, Cellina di Nardò is characterized by the highest levels of anthocyanin pigments such as cyanidin 3-O-glucoside (14.77 mg/g fw) and cyanidin-3-O-rutinoside (3.17 mg/g fw) responsible for the characteristic black color. The results indicated that fermented black table olives are an excellent natural source of bioactive compounds and can represent a potential important functional food.

DNA-based approaches were used to characterize a strain (Salento-1) of Xylella fastidiosa obtained from an olive plant suffering from the syndrome of quick decline in Apulia (South Italy). Salento-1 was indistinguishable from strain CoDiRO previously isolated from olive in Apulia and assigned to X. fastidiosa subsp. pauca. Based on our results and comparative analysis with reported data, the subspecies pauca, multiplex, and fastidiosa may invade olive throughout the world (California, Italy, Argentina and Brazil). The strain Salento-1 has been deposited in the National Collection of Plant Pathogenic Bacteria (NCPPB), England, and in the Belgian Coordinated Collections of Microorganisms (BCCM), Belgium.

Laccases are biotechnologically interesting enzymes belonging to the polyphenol oxidases family. They are widely distributed throughout the phylogenetic scale from bacteria to mammals. In fungi the analysis of three-dimensional crystal structures of laccases indicate that ascomycete laccases are processed at their C-termini, at a conserved cleavage site, resulting in the proteolytic removal of C-terminal residues. We have isolated and cloned in expression vectors the cDNAs encoding two laccase isoforms (Ery3 and Ery4) from the basidiomycete Pleurotus eryngii. The Ery3 enzyme expressed in Saccharomyces cerevisiae, is functional, whereas the recombinant Ery4 protein does not show enzymatic activity. In order to explain this evidence, we investigated the relationship between the structure of the C-terminal extension and laccase enzymatic activity. The tasks of the present study were to determine the biological role of laccase C-terminal, and to validate a "molecular engineering" approach for the production of recombinant laccases with novel biochemical properties. Genetically engineered mutant genes were produced from Ery4 by: i) progressive 3'-terminal deletions, ii) point mutations, iii) Ery3/Ery4 chimeras. The mutant genes were expressed in S. cerevisiae and active recombinant laccase isoforms were produced, exhibiting each a different biological behaviour. The correlations between the structural information deriving from both biochemical and bioinformatic analyses shed light on the role of Cterminal region in determining laccase functions. The obtained data also indicated that our approach could represent an efficient method for laccase genetic engineering. To our knowledge, this study has produced the first evidences obtained by biotechnological approach of the involvement of the C-terminal tail in the inactivation/activation process of a basidiomycete laccase.

Laccases are biotechnological relevant enzymes belonging to the group of polyphenol oxidases. Three-dimensional crystal structures analyses of fungal laccases showed that ascomycete laccases are processed at their C-termini at a conserved cleavage site, resulting in the proteolytic removal of C-terminal residues. We have isolated and cloned the cDNAs encoding the Ery3 and Ery4 laccases from Pleurotus eryngii. The Ery3 gene was functionally expressed in Saccharomyces cerevisiae, whereas the recombinant Ery4 protein did not show enzymatic activity. In order to explain this evidence, we investigated the relationship between the structure of the carboxyl-terminal extension and the enzymatic laccase activity. The tasks of the present study were to determine the biological role of laccase C-terminal and to validate a “protein engineering” approach for the production of recombinant laccases with novel biochemical properties. We produced several mutant genes derived from the ERY4 by: i) progressive 3’-terminal deletions, ii) point mutations, iii) ERY3/ERY4 chimeras. The obtained genes were expressed in S. cerevisiae and several recombinant active laccase isoforms were produced, each showing different biological behaviors. The correlations between the structural information deriving from the biochemical and bioinformatic analyses shed light on the role of laccase C-terminal region in determining laccase functions. The obtained data also indicated that the employed approach could represent an efficient method for laccase protein engineering. To our knowledge, this study has produced the first evidences of the involvement of the C-terminal tail in the inactivation/activation mechanism of a basidiomycete laccase

The ARGO-YBJ array for EAS detection at high altitude (4300 m a.s.l.) is the largest RPC-based detector in operation at present (active area of 6700 m 2 out of a total instrumented surface of about 10 4 m 2 ). The constant monitoring of the main operational and environmental parameters is essential for both keeping the detector status under control and understanding the detector behaviour more deeply. Exploiting the information provided by the ARGO-YBJ Detector Control System, the present status of the detector is described and some correlations between the environmental and the operational parameters of the ARGO-YBJ RPCs are shown. This study was performed using the monitored data from the complete ARGO-YBJ array which has been in operation since October 2007. Preliminary results on the observations of the Moon shadow and of γ -ray point sources are also presented.

The ARGO-YBJ detector at the YangBaJing Cosmic Ray Observatory (4300 m a.s.l., Tibet, P.R. China) has been put into operation in the full congration since November 2007. It is the rst EAS detector combining a very high mountain altitude with a full coverage detection surface. The high time-space granularity combined with the full coverage make ARGO-YBJ a unique device to study the EAS characteristics. In this paper we report a few selected results in Gamma-Ray Astronomy and Cosmic Ray Physics

ARGO-YBJ is the first EAS detector combining a very high mountain altitude (4300 m a.s.l.) to a "full coverage" detection surface. These features allow ARGO-YBJ to work with an energy threshold as low as a few hundreds GeV. The high duty cycle and the large field of view (~2 sr) make ARGO-YBJ suitable to monitor the gamma ray sky searching for unknown sources and unexpected events, like Active Galactic Nuclei flaring episodes or high energy Gamma Ray Bursts. In this paper we present the first observations of ARGO-YBJ concerning gamma ray astronomy, in particular the detection of the Crab Nebula and the blazar Markarian 421 during the 2006 and 2008 outbursts, and the results of a search for Gamma Ray Bursts emission in the GeV energy range using the scaler mode technique.

We report on the search for Gamma Ray Bursts (GRBs) in the energy range 1-100 GeV in coincidence with the prompt emission detected by satellites using the Astrophysical Radiation with Ground-based Observatory at YangBaJing (ARGO-YBJ) air shower detector. Thanks to its mountain location (Yangbajing, Tibet, P.R. China, 4300 m a.s.l.), active surface (about 6700 m**2 of Resistive Plate Chambers), and large field of view (about 2 sr, limited only by the atmospheric absorption), the ARGO-YBJ air shower detector is particularly suitable for the detection of unpredictable and short duration events such as GRBs. The search is carried out using the "single particle technique", i.e. counting all the particles hitting the detector without measurement of the energy and arrival direction of the primary gamma rays. Between 2004 December 17 and 2009 April 7, 81 GRBs detected by satellites occurred within the field of view of ARGO-YBJ (zenith angle < 45 deg). It was possible to examine 62 of these for >1 GeV counterpart in the ARGO-YBJ data finding no statistically significant emission. With a lack of detected spectra in this energy range fluence upper limits are profitable, especially when the redshift is known and the correction for the extragalactic absorption can be considered. The obtained fluence upper limits reach values as low as 10**{-5} erg cm**{-2} in the 1-100 GeV energy region. Besides this individual search for a higher energy counterpart, a statistical study of the stack of all the GRBs both in time and in phase was made, looking for a common feature in the GRB high energy emission. No significant signal has been detected., Comment: accepted for publication in ApJ

L'approccio biotecnologico prevede l'utilizzo di nuovi microrganismi isolati da differenti fonti con capacità di metabolizzare i composti tossici contenuti nelle AV e ridurne quindi il carico inquinante delle. I microrganismi possono essere prodotti come starter per la fermentazione e il trattamento delle AV in forma di cellule libere o immobilizzate. In tal modo si può ottenere una riduzione sensibile, dal 30 al 50% della domanda chimica di ossigeno (COD) delle acque, della concentrazione di molecole fenoliche presenti, della carica antimicrobica e fitotossica. L'approccio biotecnologico proposto potrebbe garantire nel medio periodo di ottenere delle acque con ridotta o nulla carica inquinante ed umificate, da impiegare senza limitazioni per la ferti-irrigazione dei terreni agricoli. Le AV trattate con la tecnologia proposta, rappresentano inoltre un substrato più facilmente utilizzabile da specifici ceppi batterici per la produzione di biogas.

The understanding of the yeast population dynamics during spontaneous alcoholic fermentation allows to preserve the microbial biodiversity, to use as indigenous fermentation starters so and to improve the organoleptic and sensory properties of the produced wines. However, it is similarly important to investigate the safety aspects of microbial biodiversity, in particular, on the undesired production of biogenic amines (BAs), low-molecular-weight organic bases produced in wine by the activity of microbial-specific amino acid decarboxylases. This study is the first large-scale investigation on vineyard-associated yeast strains from Apulia (Southern Italy). Eight natural must fermentations were carried out by sampling grape (Vitis vinifera) in the most significant production areas for Negroamaro and Primitivo cultivars: Torchiarolo, Copertino, Cutrofiano and Melissano for the former, Galatina, Torchiarolo, Manduria and Gioia del Colle areas for the latter. Yeasts isolates were identified by PCR ribotyping and bioinformatic analysis of the rRNA Internal region denoted as Transcribed Spacer (ITS). The Saccharomyces cerevisiae strains were further identified and differentiate as strain level by evaluating the polymorphism of their interdelta elements. The results of the molecular analyses revealed the presence of twenty different species belonging to 9 genera. In particular, Hanseniaspora uvarum, Metschnikowia pulcherrima and Aureobasidium pullulans were the dominant strains on the grape surface, whereas M. pulcherrima and H. uvarum were predominant during the early fermentation stage. We identified 692 S. cerevisiae isolates and a number of different strain in each of the 8 fermentations strains, ranging from 26 to 55, The strains were assayed for BAs production, either in synthetic media or grape must. Two Pichia manshurica, an Issatchenkia terricola and a M. pulcherrima strains were capable to produce in wine histamine and cadaverine. The production of BAs in the synthetic medium was dissimilar than that detected in wine, thus enhancing significance to assess the yeast BAs production by an "in grape must" assay, in order to reducethe potential health risk for consumer represented by these spoilage yeasts. To the best of our knowledge, this is the primary study regarding the biodiversity and safety aspects of grape-associated yeast strains in this important wine-producing area of Southern Italy.

La caratterizzazione e l'utilizzo a livello industriale di ceppi selezionati di lieviti e batteri lattici, definiti colture starter, è ormai una parte fondamentale delle biotecnologie applicate alle produzioni enologiche. Come starter sono definite le biomasse microbiche che servono a guidare un determinato processo fermentativo. I vantaggi del loro uso in enologia sono molteplici: i) consentono di condurre la fermentazione con maggiore rapidità; ii) controllano il processo evitando lo sviluppo di fermentazioni anomale ad opera di microrganismi indesiderati e/o alteranti; iii) diminuiscono le problematiche relative ai rallentamenti e agli arresti di fermentazione; iv) permettono di ottenere un prodotto finale di caratteristiche organolettiche costanti.

Table olives are one of the most important traditional fermented vegetables in Southern European countries and their consumption is constantly increasing throughout the world. Today, the industrial production of black table olives is carried out by spontaneous fermentation processes which are not predictable and are strongly influenced by the autochthonous microflora, the physical-chemical conditions, the availability of fermentable substrates and salt content. Evolution of sugars, organic acids, alcohols, mono and polyphenol compounds and volatile compounds associated with the fermentative metabolism of yeasts and bacteria throughout the natural fermentation process of the two Italian olive cultivars Cellina di Nardò and Leccino were determined. A new protocol was developed and applied aimed at the selection of LAB and yeast strains as candidate autochthonous starters for table olive fermentation from Cellina di Nardò and Leccino cultivars.

This work is the first large-scale study on vineyard-associated yeast strains from Apulia (Southern Italy). Yeasts were identified by Internal Transcribed Spacer (ITS) ribotyping and bioinformatic analysis. The polymorphism of interdelta elements was used to differentiate Saccharomyces cerevisiae strains. Twenty different species belonging to 9 genera were identified. Predominant on the grape surface were Metschnikowia pulcherrima, Hanseniaspora uvarum and Aureobasidium pullulans, whereas M. pulcherrima and H. uvarum were dominant in the early fermentation stage. A total of 692 S. cerevisiae isolates were identified and a number of S. cerevisiae strains, ranging from 26 to 55, was detected in each of the eight fermentations. The strains were tested for biogenic amines (BAs) production, either in synthetic media or grape must. Two Pichia manshurica, an Issatchenkia terricola and a M. pulcherrima strains were able to produce histamine and cadaverine, during must fermentation. The production of BAs in wine must was different than that observed in the synthetic medium. This feature indicate the importance of an "in grape must" assessment of BAs producing yeast. Overall, our results suggest the importance of microbiologicalcontrol during wine-making to reduce the potential health risk for consumer represented by these spoilage yeasts.

La fermentazione malolattica (FML) è la conversione dell'acido L-malico in acido L-lattico e CO2 attuata da parte dei batteri malolattici (BML) a seguito della loro crescita nel vino. Questo processo fermentativo provoca la disacidificazione del vino, in quanto un acido di-carbossilico, l'acido malico, viene trasformato in un acido mono-carbossilico quale l'acido lattico. Associate a questo processo avvengono altre trasformazioni importanti dal punto di vista organolettico quali modificazioni del colore, rivelazione di aromi e modificazioni dell'impatto gustativo. Essa può avvenire spontaneamente ad opera della flora indigena o mediante l'utilizzo di colture starter selezionate, solitamente appartenenti alla specie Oenococcus oeni. I vantaggi dell'induzione della FML con l'inoculo di BML selezionati sono un maggiore controllo sull'avvio e sul completamento della degradazione dell'acido malico ed un effetto positivo sull'aroma e sul gusto del vino. Il momento dell'inoculo batterico gioca un ruolo importante nella definizione del profilo sensoriale del vino . Generalmente si raccomanda l'inoculo dei batteri nel vino dopo la fermentazione alcolica (FA), quando la concentrazione di zuccheri è bassa. Infatti, una possibile conseguenza indesiderata del metabolismo eterofermentativo dei BML è lo spunto lattico ovvero la degradazione degli zuccheri con conseguente produzione di acido acetico e di acido D-lattico. In vini che al termine delle FA presentano particolari condizioni chimico-fisiche come elevato livello di etanolo, carenze nutrizionali, pH basso, elevati tenori di SO2 molecolare, rendono difficile lo sviluppo dei BML si consiglia il coinoculo (inoculo simultaneo di lieviti e batteri). Una conseguenza importante della tecnica del coinoculo è la notevole riduzioni dei tempi di fermentazione, che risulta vantaggiosa sia dal punto di vista economico sia dal punto di vista tecnico-pratico di gestione della cantina e riduce inoltre il rischio di alterazioni microbiologiche dei vini prodotti . Va comunque detto che la pratica della coinoculazione è un'operazione molto delicata, da gestire con cura ed estrema attenzione [6] e sulla quale poche informazioni sono disponibili in letteratura.Scopo di questo lavoro sperimentale è stato quello di studiare gli effetti a livello fisiologico e chimico dell'inoculo simultaneo in mosto d'uva del lievito Saccharomyces cerevisiae e del batterio O. oeni, per lo svolgimento in contemporanea delle fermentazioni alcolica e malolattica per una sua applicazione in condizioni difficili di vinificazione, ovvero, ovvero alto tenore alcolico e elevata acidità, tipiche dell'enologia Meridionale.

Spontaneous grapes must fermentations are promoted by the indigenous yeasts, that are able to confer a distinctive style and quality to the produced wine. The spontaneous fermentations of grape must are at first dominated by non-Saccharomyces yeasts and, in a final stage the alcoholic fermentation process is completed by dominant S. cerevisiae strains (Bauer and Pretorius, 2000).The autochthonous yeast strains are associated to a specific vineyard niche habitat and their role in natural fermentation allows the production of wines with particular features in each microclimatic area (Pérez-Coello et al., 1999). However, in order to avoid the unpredictability of must spontaneous fermentation, the winemakers employ commercial dry active yeast culture for wine industrial productions. Increasing interest in the application of locally selected yeasts for fermentation management has been reported (Tristezza et al., 2012). The employment of autochthonous strains of S. cerevisiae as starters seems to be preferable since they are adapted to all the constraints related to a specific wine-production area (Lopes et al., 2007) and are thus capable to dominate more efficiently the indigenous microflora during the fermentation process. Moreover, autochthonous yeast strains can assure the preservation and/or the enhancement of the typical oenological and sensory features which could be considered representative of an oenological region (Rodríguez et al., 2010). In the present study, we developed and applied a strategy to select S. cerevisiae strains from a larger number of yeast isolates. This was achieved adopting a number of key parameters indicative of the strains technological and enological properties. S. cerevisiae population has been isolated from natural fermentations of grape musts, which derive from grapes sampled from the six most representative Negroamaro and Primitivo producing-areas in Apulia (Southern Italy). The yeast populations were identified by molecular assays (AFLP and sequencing) and some selected representative strains were subjected to physiological, oenological and technological characterization. At the end of the selection procedure, that lasted three years, three indigenous S. cerevisiae strains (one for Primitivo and two for Negroamaro), characterized by interesting technological and oenological properties, were selected. The three selected strains were evaluated by both laboratory tests and semi-industrial scale fermentations to confirm their ability to act as autochthonous fermentation starters. An optimized procedure was worked out for the production of starter biomasses, in order to test them in Negroamaro and Primitivo wine production, at an industrial scale, in six different wineries. The employment of autochthonous starter cultures for the industrial production of typical wines in Apulia will be discussed.

Two approaches were developed in order to select microorganisms suitable to be used in olive mill wastewaters bioremediation. By the first approach, three hundred yeasts were isolated from five industrial mills and identified by molecular analysis. The different strains were selected according to their capacity to grow in OMW (olive mill wastewaters) as the sole carbon source and to reduce phenolics, chemical oxygen demand (COD) and antimicrobial compounds. One Geotrichum candidum isolate was used to set up a whole-cell immobilization system in calcium alginate gel and the COD and phenolic reduction obtained using the immobilized cells showed respectively a 2.2- and 2-fold increase compared to the removal obtained using free cells. By the second approach, a new protocol was developed to isolate and select aerobic microorganisms from different industrial samples and environmental niches (soils, OMWs) and able to detoxify olive mill wastewaters.

Apulia is the second Italian wine-producing region and the most important producer of red-rosé wine. One peculiar characteristic of Apulian wines is that the podologic characteristics and the climatic conditions of this region contribute to enrich the wine of aromatic essences and to give, therefore, a characteristic and intense taste to the finished product. The Apulian wine industry is living a moment of great qualitative transformation and the challenge of the market had therefore addressed the productive row towards the use of innovative systems to guarantee and exalt the qualitative characteristics of regional wines. A heartfelt requirement is to being able to pilot and to control the productive activity to obtain wines with peculiar characteristics, with respect of the typicality, that is guaranteed by the denominations of origin. The selection and the employment of new combinations of microorganisms, obtained from the native micro flora would be a powerful instrument to improve the organoleptic and sensory characteristics of the product. Autochthonous yeasts are the micro-organisms better adapted to a specific must, which detain characteristics determined by the variety of the grapes and the terroir and, therefore they are able to exalt the peculiarities (aromas, structure and colour) of a wine. In these last year the research activity of the Institute of Science of Food Production of C.N.R. has been directed to the exploitation of autochthonous microbiota to enhance the quality of regional wines. This research line has been granted by a number of National and Regional Projects and it has been conducted in collaboration between the ISPA and a large number of wine companies. The research activity has generated a virtuous circle, thus allowing the standardization of protocols for wine yeast enological selection and biomass production, the constitution of a Yeast Collection but, most of all, the transfer of technology to a large number of SMEs belonging of the wine production chain. Natural fermentations of Negroamaro, Primitivo and Susumaniello musts have been performed and the Saccharomyces cerevisiae population has been analyzed and characterized by molecular, physiological, enological and technological tests allowing the identification of four indigenous S. cerevisiae strains candidate as autochthonous fermentation starters. The enological properties of the above strains have been evaluated during the vintages 2006-2009, by performing 37 large scale vinification trials in 21 different industrial cellars in Apulia. The obtained results and their implications for the valorisation and the improvement of Apulian typical wines will be discussed.

The wine is the product of the spontaneous grape must fermentation, induced by the indigenous micro flora, which catalyse the rapid, complete and efficient conversion of grape sugar into ethanol, carbon dioxide and other minor, but important metabolites by the complex biochemical process denoted as "alcoholic fermentation". The first step of the fermentation is characterized by the growth of yeast belonging to Kloeckera, Hanseniaspora, Candida, Metschnikowia e Pichia genera. These yeasts produce very low amount of alcohol but, they generate during the fermentation a large number of other secondary metabolites, which could positively influence the final bouquet of the wine. When the alcohol concentration raise to 4%, the second step of the fermentation process begins, which is mainly dominated by Saccharomyces cerevisiae. This yeast, commonly denoted as the "wine yeast", completes the fermentation, with high alcohol quantity but low production of secondary metabolites. The present study was aimed to the molecular and technological characterization of autochthonous yeast strains belonging to the Hanseniaspora uvarum species, isolated from spontaneous fermentation of Negroamaro and Primitivo grapes. The criterion for H. uvarum strain differentiation was provided by the PCR analysis of Random Amplified Polymorphic DNA (RAPD) markers and this assay which already demonstrated to allow the differentiation of yeast strains. The identified strains have been employed to fermentation laboratory tests after their inoculation in grape must and their oenological and technological properties have been evaluated. The fermented musts were further analyzed by high pressure liquid chromatography (HPLC) and gas-chromatography assays. The results of these molecular and technological analyses for the characterization of autochthonous H. uvarum strains will be discussed.

La fermentazione malolattica (FML) è un evento fermentativo successivo alla fermentazione alcolica compiuta dai batteri lattici, in particolare da Oenococcus oeni. La FML influenza la qualità del vino ed è considerata, per questo, un evento importante nel processo di vinificazione. Tale processo avviene sia spontaneamente ad opera della flora indigena o artificialmente mediante inoculo di colture starter. Quest'ultimo processo è una pratica comunemente utilizzata in cantina e le fermentazioni condotte con l'inoculo di ceppi batterici selezionati sono più facili da controllare e garantiscono lo standard qualitativo del prodotto. Scopo di questo studio é stata la caratterizzazione biomolecolare e tecnologica di ceppi isolati da mosto di uve Salentine "Primitivo" tipica cultivar della Puglia. Durante le differenti fasi del processo di FML (inizio, media e fine) è stata isolata una popolazione di 220 isolati di batteri lattici, da cui sono stati identificati 87 ceppi di Oenococcus oeni. L'identificazione è stata effettuata mediante analisi PCR specie-specifica utilizzando oligonucleotidi specifici disegnati per il gene MLE e successivo sequenziamento dei prodotti di PCR relativi all'RNA contenente la regione per il gene 16S rRNA. Questo ha permesso di ottenere delle sequenze discriminanti le diverse specie. L'analisi genotipica è stata eseguita con lo sviluppo della tecnica molecolare AFLP, per la prima volta adottata sul genoma di questo batterio, al fine di sviluppare profili molecolari discriminanti che differenziano e caratterizzano gli Oenococcus oeni a livello di ceppo. In seguito a questa analisi è stato possibile differenziare quattro differenti gruppi di isolati, tre dei quali hanno mostrato significativi livelli di omologia intraspecifica. L'analisi AFLP, si è dimostrata una valida alternativa ad altri metodi molecolari. Un totale di 28 ceppi, scelti dai differenti sottogruppi dei cluster ottenuti dagli AFLP, sono stati testati per la loro capacita di metabolizzare il malato. Significative differenze sono state osservate tra i ceppi relativamente alla a) velocità del consumo di acido malico, b) biomassa prodotta e al c) tasso specifico di consumo di acido malico. I risultati ottenuti e la loro implicazione biotecnologia verranno discusse.

One hundred and fourty-four strains of epiphytic micro-organisms, isolated from berries of "Negroamaro ", an apulian grape wine variety, were screened for antagonistic activity against Aspergillus carbonarius and A. niger. These black Aspergilli are considered the main ochratoxigenic species responsible for the accumulation of ochratoxin A in grape. Twenty eight yeast isolates were selected for their inhibitory effects on the above fungal species and assayed by an in vitro nutritional competition test for their antagonistic capacity towards three selected ochratogenic strains. Six yeast isolates belonging to five species, namely 2 isolates of Issatchenkia orientalis and one each of Metschnikowia pulcherrima, Kluyveromyces thermotolerans, Issatchenkia terricola and Candida incommunis species, were finally selected and screened on wounded grape berries for their ability to inhibit the contemporary infection by ochratoxigenic molds and reduce the formation of ochratoxin A. With the exception of the K. thermotolerans isolate, when inoculated at 109 CFU/wound, the other five challenger yeasts reduced the A. carbonarius and A. niger colonization on grape berry (P < 0.05). In particular, the best antagonistic activity was shown by the two I. orientalis isolates. Results suggest that antagonist yeasts with the potential to control of A. carbonarius and A. niger on grape can be found among the microflora associated with the berries.