APPLICATIONS OF BIOMOLECULAR METHODS ALONG THE PDO WINES CHAIN PRODUCTION

Il presente progetto di dottorato ha cercato di rispondere a questioni, esigenze e preoccupazioni riguardanti l'intera produzione della filiera vitivinicola. In particolare, si è articolato in due filoni di ricerca principali, strettamente integrati tra loro. La prima parte ha riguardato l'applicazione dell’analisi del DNA per la tracciabilità delle produzioni vinicole di qualità, utilizzando banche dati di profili SSR per l’attribuzione varietale ad uve, mosti e vini come VIVC - Vitis International Variety Catalogue, VitisDB -Italian Vitis Database, ecc. Per consolidare il lavoro di tracciabilità, in un secondo momento, si è prestata particolare attenzione alla delicata fase di conservazione del vino, al fine di valutare per quanto tempo fosse possibile tracciare le produzioni enologiche durante la fase di pre-imbottigliamento e vino confezionato. Mentre, la seconda parte del lavoro, ha avuto come focus l’analisi metagenomica della diversità batterica e la profilazione metabolomica di mosti (in varie fasi di lavorazione) e vini di Buttafuoco Storico prodotti, considerando diversi regimi colturali, in un'area produttiva di nicchia dell'Oltrepò Pavese. Conoscere i meccanismi d'azione dei microrganismi coinvolti e capire come questi possano influenzare positivamente o negativamente la qualità del prodotto finito è di grande aiuto per arrivare alla bottiglia con il miglior risultato.The present doctoral project has attempted to respond to the issues, needs and concerns regarding the entire wine chain production. In particular, it consisted of two main, closely integrated research strands. The first part dealt with the application of DNA analysis for the traceability of quality wine production by using databases of SSR profiles used as references for grapevine varietal identification as VIVC - Vitis International Variety Catalogue, VitisDB -Italian Vitis Database, etc (recognition of varieties used for wine production). To consolidate the traceability work, special attention was paid to the delicate phase of wine storage in order to assess how long it was possible to trace the productions in the case of pre-bottling stage and packaged wine. While the second part of the work focused on metagenomic bacterial diversity and metabolomics profiling of Buttafuoco Storico musts (at various stages of processing) and wines production, considering different cultural regimes, in a niche production area of Oltrepò Pavese. Knowing the mechanisms of action of the microorganisms involved and understanding how they can positively or negatively influence the quality of the finished product would be a great help in getting to the bottle with the best result

Scalable synthesis and purification of functionalized graphene nanosheets for water remediation.

PEI modified graphene oxide (GOPEI) is prepared by tandem MW amination–microfiltration and used for arsenic removal from water

The Rediscovery of Traditional Maize Agrobiodiversity: A Study Case from Northern Italy

Nowadays, agriculture is under the pressure of climate change and new pathogen outbreaks while farmers are requiring breeders to develop more resistant and resilient genotypes. The genetic base for breeding may be increased through appropriate conservation, description and characterization of local varieties and germplasm collections that have never been used in breeding, and which could be sources of useful alleles. In this framework, the present paper focuses on eight maize landraces of the eastern part of Emilia-Romagna, derived from the Italian maize collection sampled in 1954. Landraces are characterized by a short cycle length and different kernel types—mainly flint-like or an intermediate type of yellow or yellow–orange color—while dent landraces are less represented. Pigmented and white corns are absent even though one landrace (Va213) showed the presence of scattered blue kernels on yellow ears. Ear shape is frequently conical, a trait associated with drought-resistance and common in Italian traditional landraces. Genetic characterization was carried out on 529 individuals by using 10 SSR markers. A total of 68 different alleles, ranging from 4 for markers (phi084 and umc1401) to 11 (phi031) and from 27 (Va217) to 50 (Va211), were evidenced at the individual and population level. AMOVA analysis revealed a small amount (19%) of variability between populations, as supported also by PCoA, with the only exception of Va217, which is different from the others, as evidenced also by phylogenetic analysis. Population structure analysis resulted in the identification of three and four population levels, which are consistent with previous results

Graphene oxide-polysulfone hollow fibers membranes with synergic ultrafiltration and adsorption for enhanced drinking water treatment

Polysulfone-graphene oxide hollow fiber membranes (PSU-GO HFs) with simultaneous adsorption and ultrafiltration capabilities are herein described and proposed for enhanced and simplified Point-of-Use (POU) drinking water purification. The PSU-GO HFs were prepared by phase inversion extrusion by a customized semi-industrial plant and their morphology, surface properties, and porosity were investigated by combined Scanning Electron Microscopy (SEM), contact angle and Raman confocal microscopy, in relation to different GO:PSU ratios (1–5% w/w GO vs PSU) and to the final adsorption-ultrafiltration properties. Filtration modules of PSU-GO HFs of filtering surface (FS) in the range 0,015–0,28 m2 showed same ultrafiltration capability of PSU-HF standard filters. Synergic adsorption properties were demonstrated by studying the adsorption maximum capacity of ciprofloxacin antibiotic (CIPRO) vs GO ratio in dead end in-out configuration, the standard configuration used for PSU HFs commercial modules. Loading of 3,5% GO vs PSU was selected as case study, representing the best compromise between performance and GO nanofiller amount. Heavy metals (Pb, Cu and Cr(III)) and polyfluoroalkyl substances (PFAS) removal capabilities from tap water were competitive and in some cases outperformed Granular Activated Carbon (GAC), the standard industrial sorbent. Ciprofloxacin removal from tap water was also under real operational conditions. Moreover, release of GO from working PSU-GO modules was excluded by Surface Enhanced Raman Spectroscopy (SERS) analysis of treated water having the state-of-the-art limit of quantification of 0.1 μg/L for GO nanosheets

Defective graphene nanosheets for drinking water purification: Adsorption mechanism, performance, and recovery

Defect-rich graphene oxide (dGO) was used as sorbent for organic contaminants of emerging concern in tap water, including drugs and dyes, and the performance compared to those of lower-defects graphene types. The role of holes and carbonyl- carboxylic groups on graphene nanosheets surface on the adsorption mechanism and efficiency was investigated. dGO showed enhanced adsorption capacity toward two fluoroquinolone antibiotics (ofloxacin, OFLOX, and ciprofloxacin, CIPRO) in tap water with a maximum capacity of 650 mg/g, compared to 204 mg/g for Hummers derived commercial GO (hGO) and 125 mg/g for less defected Brodie derived GO (bGO) for OFLOX. The role of defects on the selective adsorption of OFLOX was also modelled by MD simulations, highlighting a mechanism mainly driven by the shape complementarity between the graphene holes and the molecules. Adsorption isotherms revealed different adsorption model for dGO, with a Langmuir fitting for dGO and BET fitting for all the other investigated samples. The maximum adsorption capacity of dGO for OFLOX was about six times higher than that of Granular Activated Carbon (95 mg/g), the industrial adsorption standard technology. Finally, it was also demonstrated that dGO can be recovered from treated water by ultrafiltration, this preventing secondary contamination risks and enabling safe use of graphene nanosheets for water purification

Multicolor, large-area fluorescence sensing through oligothiophene-self- assembled monolayers

We present a new strategy to realize self-assembled monolayers (SAMs) on quartz and silicon with a multicolour fluorescence pattern starting from a single, proton sensitive oligothiophene dye exposed at a defined pH. Fine tuning of the SAMs emission color over the entire visible range, including white, is demonstrated. Finally, integration of SAMs in patterned thin layer cells (TLCs) is exploited to demonstrate cation sensing potential in real devices. Keyword

Facile tuning from blue to white emission in silica nanoparticles doped with oligothiophene fluorophores

Oligothiophenes (TFs) with blue, green and orange emission have been used for the first time as doping fluorophores of silica nanoparticles (SiO(2)NPs). High purification of the new nanoparticles (TFsSiO(2)NPs) from free molecular fluorophores was achieved by means of asymmetrical flow field-flow fractionation on-line combined with multi-angle light scattering and fluorescent detection (AF4-MALS-FD). The synthesis, structural, compositional and optical characterizations of the new TFsSiO(2)NPs are reported. We show that the tailored co-assembly of TFs in bi- and tricomponent TFsSiO(2)NPs allows for the fine-tuning of the emission of the nanoparticles from blue to white by means of FRET processes between adjacent TFs. These unique optical signatures make TFsSiO(2)NPs potentially effective tools for fluorescent sensing and labeling

Effect of storage time on wine DNA assessed by SSR analysis

Wine traceability based on DNA analysis has been reported in several previous studies but no one until now had monitored how far molecular traceability was possible during wine storage, before and after bottling. The present study tries to fill this gap of knowledge by following, in the real case of a large wine cooperative, the possibility to trace wine production through DNA analysis during storage period. Two monovarietal productions: red sparkling Bonarda PDO and white Pinot gris PDO, were followed starting from the end of oenological practices until 1 year after bottling. During the pre-bottling period, samples were collected every 10 days during four consecutive months, after bottling samples were collected at day 1 and after 2, 8 and 12 months. DNA analysis evidences that for both wines, traceability by applying SSR analysis on the extracted DNA is possible, at least, until month 8, after that DNA degradation increases hindering, mainly for the red wine, the possibility to a correct varietal identification

Defective graphene nanosheets for drinking water purification : Adsorption mechanism, performance, and recovery

Defect-rich graphene oxide (dGO) was used as sorbent for organic contaminants of emerging concern in tap water, including drugs and dyes, and the performance compared to those of lower-defects graphene types. The role of holes and carbonyl- carboxylic groups on graphene nanosheets surface on the adsorption mechanism and efficiency was investigated. dGO showed enhanced adsorption capacity toward two fluoroquinolone antibiotics (ofloxacin, OFLOX, and ciprofloxacin, CIPRO) in tap water with a maximum capacity of 650 mg/g, compared to 204 mg/g for Hummers derived commercial GO (hGO) and 125 mg/g for less defected Brodie derived GO (bGO) for OFLOX. The role of defects on the selective adsorption of OFLOX was also modelled by MD simulations, highlighting a mechanism mainly driven by the shape complementarity between the graphene holes and the molecules. Adsorption isotherms revealed different adsorption model for dGO, with a Langmuir fitting for dGO and BET fitting for all the other investigated samples. The maximum adsorption capacity of dGO for OFLOX was about six times higher than that of Granular Activated Carbon (95 mg/g), the industrial adsorption standard technology. Finally, it was also demonstrated that dGO can be recovered from treated water by ultrafiltration, this preventing secondary contamination risks and enabling safe use of graphene nanosheets for water purification

Metagenomic bacterial diversity and metabolomics profiling of Buttafuoco wine production

Buttafuoco dell’Oltrepo’ Pavese (or Buttafuoco) is an important and renowned red wine, protected by a Denominazione di Origine Controllata (DOC) designation established in 2010, produced in the Northern Italy in the province of Pavia (Italy). The knowledge of factors as the typical microbial terroir and the metabolite composition of the wine is fundamental for producing excellent wines. In this work, two productions of Buttafuoco Storico dell’Oltrep`o Pavese were followed in order to assess the microbial populations through different stages of the wine production chain and the metabolomic composition of the final wines. Microbial terroir was investigated through a metagenomic analysis that revealed a wide microbial consortium which is, for the major taxonomic groups, affected by sampling time over location. Before the metagenomic analysis, being DNA extraction from wine a difficult task, two different approaches were compared for a precise quantification of microbial DNA (bacteria and yeast): digital and real time PCR. Obtained results clearly evidenced that digital PCR being was more sensitive than real time PCR and likely the method of choice for quantifying DNA extracted from processed matrices. Metabolomic profiling, focused on phenolic compounds, was able to clearly distinguish among vineyards and to highlight the presence of discriminating molecules that can be related to the different edaphic conditions