In the last decades, the increase of industrial activities and of the request for the world food requirement, the intensification of natural resources exploitation, directly connected to pollution, have aroused an increasing interest of the public opinion towards initiatives linked to the regulation of food production, as well to the institution of a modern legislation for the consumer guardianship. This work was planned taking into account some important thematics related to marine environment, collecting and showing the data obtained from the studies made on different marine species of commercial interest (Chamelea gallina, Mytilus edulis, Ostrea edulis, Crassostrea gigas, Salmo salar, Gadus morhua). These studies have evaluated the effects of important physic and chemical parameters variations (temperature, xenobiotics like drugs, hydrocarbons and pesticides) on cells involved in the immune defence (haemocytes) and on some important enzymatic systems involved in xenobiotic biotransformation processes (cytochrome P450 complex) and in the related antioxidant defence processes (Superoxide dismutase, Catalase, Heat Shock Protein), from a biochemical and bimolecular point of view. Oxygen is essential in the biological answer of a living organism. Its consume in the normal cellular breathing physiological processes and foreign substances biotransformation, leads to reactive oxygen species (ROS) formation, potentially toxic and responsible of biological macromolecules damages with consequent pathologies worsening. Such processes can bring to a qualitative alteration of the derived products, but also to a general state of suffering that in the most serious cases can provoke the death of the organism, with important repercussions in economic field, in the output of the breedings, of fishing and of aquaculture. In this study it seemed interesting to apply also alternative methodologies currently in use in the medical field (cytofluorimetry) and in proteomic studies (bidimensional electrophoresis, mass spectrometry) with the aim of identify new biomarkers to place beside the traditional methods for the control of the animal origin food quality. From the results it’s possible to point out some relevant aspects from each experiment: 1. The cytofluorimetric techniques applied to O. edulis and C. gigas could bring to important developments in the search of alternative methods that quickly allows to identify with precision the origin of a specific sample, contributing to oppose possible alimentary frauds, in this case for example related to presence of a different species, also under a qualitative profile, but morpholgically similar. A concrete perspective for the application in the inspective field of this method has to be confirmed by further laboratory tests that take also in account in vivo experiments to evaluate the effect in the whole organism of the factors evaluated only on haemocytes in vitro. These elements suggest therefore the possibility to suit the cytofluorimetric methods for the study of animal organisms of food interest, still before these enter the phase of industrial working processes, giving useful information about the possible presence of contaminants sources that can induce an increase of the immune defence and an alteration of normal cellular parameter values. 2. C. gallina immune system has shown an interesting answer to benzo[a]pyrene (B[a]P) exposure, dose and time dependent, with a significant decrease of the expression and of the activity of one of the most important enzymes involved in the antioxidant defence in haemocytes and haemolymph. The data obtained are confirmed by several measurements of physiological parameters, that together with the decrease of the activity of 7-etossi-resourifine-O-deetilase (EROD linked to xenobiotic biotransformation processes) during exposure, underline the major effects of B[a]P action. The identification of basal levels of EROD supports the possible presence of CYP1A subfamily in the invertebrates, still today controversial, never identified previously in C. gallina and never isolated in the immune cells, as confirmed instead in this study with the identification of CYP1A-immunopositive protein (CYP1A-IPP). This protein could reveal a good biomarker at the base of a simple and quick method that could give clear information about specific pollutants presence, even at low concentrations in the environment where usually these organisms are fished before being commercialized. 3. In this experiment it has been evaluated the effect of the antibiotic chloramphenicol (CA) in an important species of commercial interest, Chamelea gallina. Chloramphenicol is a drug still used in some developing countries, also in veterinary field. Controls to evaluate its presence in the alimentary products of animal origin, can reveal ineffective whereas the concentration results to be below the limit of sensitivity of the instruments usually used in this type of analysis. Negative effects of CA towards the CYP1A- IPP proteins, underlined in this work, seem to be due to the attack of free radicals resultant from the action of the antibiotic. This brings to a meaningful alteration of the biotransformation mechanisms through the free radicals. It seems particularly interesting to pay attention to the narrow relationships in C. gallina, between SOD/CAT and CYP450 system, actively involved in detoxification mechanism, especially if compared with the few similar works today present about mollusc, a group that is composed by numerous species that enter in the food field and on which constant controls are necessary to evaluate in a rapid and effective way the presence of possible contaminations. 4. The investigations on fishes (Gadus morhua, and Salmo salar) and on a bivalve mollusc (Mytilus edulis) have allowed to evaluate different aspects related to the possibility to identify a biomarker for the evaluation of the health of organisms of food interest and consequently for the quality of the final product through 2DE methodologies. In the seafood field these techniques are currently used with a discreet success only for vertebrates (fishes), while in the study of the invertebrates (molluscs) there are a lot of difficulties. The results obtained in this work have underline several problems in the correct identification of the isolated proteins in animal organisms of which doesn’t currently exist a complete genomic sequence. This brings to attribute some identities on the base of the comparison with similar proteins in other animal groups, incurring in the possibility to obtain inaccurate data and above all discordant with those obtained on the same animals by other authors. Nevertheless the data obtained in this work after MALDI-ToF analysis, result however objective and the spectra collected could be again analyzed in the future after the update of genomic database related to the species studied. 4-A. The investigation about the presence of HSP70 isoforms directly induced by different phenomena of stress like B[a]P presence, has used bidimensional electrophoresis methods in C. gallina, that have allowed to isolate numerous protein on 2DE gels, allowing the collection of several spots currently in phase of analysis with MALDI-ToF-MS. The present preliminary work has allowed therefore to acquire and to improve important methodologies in the study of cellular parameters and in the proteomic field, that is not only revealed of great potentiality in the application in medical and veterinary field, but also in the field of the inspection of the foods with connections to the toxicology and the environmental pollution. Such study contributes therefore to the search of rapid and new methodologies, that can increase the inspective strategies, integrating themselves with those existing, but improving at the same time the general background of information related to the state of health of the considered animal organism, with the possibility, still hypothetical, to replace in particular cases the employment of the traditional techniques in the alimentary field.

O presente trabalho visa à realização de um estudo sobre a tutela jurisdicional ao direito a alimentos, com a preocupação de investigar o roteiro processual a seguir no tocante ao resguardo de uma relação jurídica vital à existência digna, e, portanto, ao próprio direito à vida, com o qual estabelece uma relação na condição de direito garantia. A mirada sobre o tema se realiza sob a perspectiva das relações familiares, conquanto essa espécie de direito material também possa surgir em outras esferas responsabilidade civil ou relação contratual -, dadas as circunstâncias sempre particulares da vida familiar, ambiente onde o Estado só está autorizado a penetrar em caráter excepcional, mesmo pela via jurisdicional, dadas as delicadas relações internas entre seus integrantes, apoiadas de regra na frágil dimensão do afeto, cujo resguardo é imperativo. O campo das relações obrigacionais alimentares, de sabida origem no princípio da solidariedade (CF, art. 3º, I), e, por decorrência, no postulado da solidariedade familiar, é campo fértil a dissidências e a hipótese do inadimplemento está sempre a reclamar do Estado uma tutela jurisdicional efetiva, ao mesmo tempo em que suficientemente congruente com os objetivos alinhados ao superior interesse da família, dilema técnico repercutido na seara do processo, que há , portanto, de ajustar técnicas, procedimento e posturas de dues sujeitos, de forma a atender conveniente e eficazmente todos os interesses envolvidos com a questão. Nesse espírito, a proposta do presente trabalho é promover uma avaliação, ainda que sobre o essencial apenas, da tutela jurisdicional realizável por meio da Lei de Alimentos, a Lei Federal nº 5478/68, tanto sob o prisma da tutela cognitiva, como da tutela executiva, examinando em cada caso os aspectos mais relevantes, as virtuais controvérsias, e as soluções mais ajustadas a um modelo constitucional de processo civil, e, por que não dizer, a um modelo de processo efetivo de família. O que se procura evidenciar, dentro dessa perspectiva, é a problemática particular das relações familiares, tanto quanto a urgência em se modelar um processo civil que lhe seja apropriado, baseado na ampliação da legitimidade das deliberações, via participação mais aguda de seus sujeitos e, sobretudo, levando em consideração um gerenciamento do processo pelo magistrado de modo a não excluir, a qualquer instante, as técnicas parajurisdicionais de solução de conflito, abrindo-se espaço efetivo à conciliação, à mediação, mesmo na pendência da demanda. / Questo lavoro há per scopo la realizzazione di uno studio sulla tutela giurisdizionale del diritto agli alimenti, preoccupandosi di investigare l´iter processuale da osservare relativamente alla protezione di un rapporto giuridico vitale ad uma esistenza degna, e, pertanto, allo stesso diritto alla vita, com cui instaura uma relazione nella condizione di diritto-garanzia. La messa a fuoco del tema avviene sotto la prospettiva dei rapporti familiari, sebbene questo tipo di diritto materiale possa sorgere in altre sfere responsabilità civile o rapporti contrattuali -, date le circostanze sempre particolari della vita familiare, ambiente in cui lo Stato solamente è autorizzato ad entrare a carattere eccezionale, anche in via giurisdizionale, dati i delicati rapporti interni che sussistono tra i suoi partecipanti, che si appoggiano normalmente nella fragile dimensione dell´affetto, il cui rispetto è imperativo. Il campo dei rapporti obbligazionari alimentari, che hanno la loro chiara origine nel principio di solidarietà (Costituzione Federale, art. 3º , I), e, da quì, nel postulato della solidarietà familiare, è campo fertile di discordie e l´ipotesi di inadempimento sempre esige dallo Stato uma tutela giurisdizionale effettiva, Che sia allo stesso tempo coerente com gli obbiettivi consoni con il superiore interesse della famiglia, dilemma tecnico, questo, che si riflette nel campo del processo, il quale dovrà adattare tecniche, procedimenti e modi di essere di due soggetti, in modo da soddisfare convenientemente e efficacemente tutti gli interessi coinvolti nel caso. In tale spirito, la proposta del presente lavoro è promuovere uma valutazione, anche se appena sull´essenziale, della tutela giurisdizionale realizzabile attraverso la Legge degli Alimenti, la Legge Federale n.5478/68, tanto sotto il prisma della tutela cognitiva, quanto della tutela esecutiva, esaminando in ciascun caso gli aspetti più rilevanti, le possibili controversie, e le soluzioni che più si adattano ad um modello costituzionale di processo civile, e, si dica pure, ad um modello di processo effettivo della famiglia. Ciò che si cerca di mettere in evidenza, in tale prospettiva, è la problematica particolare dei rapporti familiari, tanto riguardo all´urgenza di plasmare um processo civile che gli sia appropiato, basato nell´ampliamento della legittimità delle deliberazioni, attraverso la partecipazione più effettiva dei suoi soggetti e, soprattutto, prendendo in considerzione uma gestione del processo da parte del magistrato in modo da non escludere, in nessun momento, le tecniche paragiurisdizionali di soluzione del conflitto, aprendosi spazio effettivo alla conciliazione, alla mediazione, anche in pendenza di giudizio.

Grape (Vitis vinifera L.) is among the most cultivated plants in the world. Its origin traces back to the Neolithic era, when the first human communities started to domesticate wild Vitis sylvestris L. grapes to produce wines. Domestication modified Vitis vinifera to assume characteristics imparted from the humans, selecting desired traits (e.g. specific aromas), and excluding the undesired ones. This process made this species very different from all the other wild grape species existing around the world, including its progenitor, Vitis sylvestris. Metabolomics is a field of the sciences that comparatively studies the whole metabolite set of two (or more) groups of samples, to point out the chemical diversity and infer on the variability in the metabolic pathways between the groups. Crude metabolomics observation can be often used for hypotheses generation, which need to be confirmed by further experiments. In my case, starting from the grape metabolome project (Mattivi et al. unpublished data), I had the opportunity to put hands on a huge dataset built on the berries of over 100 Vitis vinifera grape varieties, tens of grape interspecific hybrids and few wild grape species analyzed per four years; all included in a single experiment. Starting from this data handling, I designed specific experiments to confirm the hypotheses generated from the observation of the data, to improve compound identification, to give statistical meaning to the differences, to localize the metabolites in the berries and extrapolate further information on the variability existing among the grape genus. The hypotheses formulated were two: 1) several glyco-conjugated volatiles can be detected, identified and quantified in untargeted reverses-phase liquid chromatography-mass spectrometry; 2) The chemical difference between Vitis vinifera and wild grape berries is wider than reported in literature. Furthermore, handling a huge dataset of chemical standards injected under the same conditions of the sample set, I also formulated a third hypothesis: 3) metabolites with similar chemical structures are more likely to generate similar signals in LC-MS, therefore the combined use of the signals can predict the more likely chemical structure of unknown markers. In the first study (chapter 5), the signals putatively corresponding to glycoconjugated volatiles have been first enclosed in a specific portion of the temporal and spectrometric space of the LC-HRMS chromatograms, then they have been subjected to MS/MS analysis and lastly their putative identity have been confirmed through peak intensity correlation between the signals measured in LC-HRMS and GC-MS. In the second study (chapter 6), a multivariate regression model has been built between LC-HRMS signals and the substructures composing the molecular structure of the compounds and its accuracy and efficacy in substructure prediction have been demonstrated. In the third study (chapter 7), I comparatively studied some wild grapes versus some Vitis vinifera varieties separating the basic components of the grape berry (skin, flesh and seeds), with the aim to identify all the detected metabolites that differentiate the two groups, which determine a difference in quality between the wild versus domesticated grapes, especially regarding wine production.

In the recent years, the consumer choices have been focused on health-promoting plant-based food and their preferences are oriented towards regional foodstuff from local productions. Therefore, an important factor for vegetables grown Trentino-Alto Adige (Italy) is to point out the added value of alpine farming to evaluate the nutritional values of farming products. Omics technologies (e.g. genomics, transcriptomics, proteomics and metabolomics) are aimed at investigating the assessment of different pools of molecules and how they are translated into the structure, function, and dynamics of a biological system or systems in order to provide a comprehensive characterization of a specific organism. Research use the omics techniques to exhaustively understand the functionality of food components. Several sophisticated chromatographic methods, spectroscopic techniques and chemometric tools are applied to give an insight into a comprehensive overview of the intrinsic quality, typicality and regionality of specific plant-based foods in the present PhD thesis: apples and potatoes. The quality of these foods is evaluated by quantifying the secondary metabolites to investigate their nutraceutical values. The aim of this PhD project is to use several analytical techniques (LC-MS, UV-VIS) that are capable of comprehensively characterizing the food metabolome with particular emphasis on those components with high nutritional values. The data analysis and data handling of omics data requires advanced bioinformatic, statistical, and chemometric tools. Potatoes and apples are chosen as target matrices for these studies for their relevance in the local economy and for the peculiar chemical composition of particular interest for their health-promoting proprieties. The information is acquired using several sophisticated chromatographic and spectroscopic techniques, such as ultra-high performance liquid chromatography coupled with triple quadrupole mass spectrometry (UHPLC– MS/MS) and UV/VIS. It is integrated to chemometric approaches (principal component analysis (PCA), partial least square regression (PLS), and data fusion) to achieve a comprehensive targeted chemical characterization. The sampling procedures gathers, in the case of the potatoes study, reference cultivars that may be found in the common retailers of Trentino/Alto-Adige and different production areas, the apples of 22 cultivars were harvest from the fields of the Laimburg Research Centre (Vadena, Italy) to guaranty comparability of the obtained data. Our results may be used as solid foundation for a reliable evaluation of apples and potatoes healthy "potential" value based on cutting-edge techniques, which are capable of providing comprehensive data regarding the alpine food quality parameters with high efficiency and reliability

Il presente studio affronta il tema della rintracciabilità alimentare di prodotti agroalimentari, analizzando il caso del pomodoro da industria, cercando di trovare degli indicatori chimici (markers) che siano in grado di discriminare prodotti provenienti da aree geografiche differenti. Viene quindi affrontato il tema della tipicità che consiste nel legame esistente tra l'alimento e il territorio nel quale viene prodotto. I parametri chimici che vengono indagati sono elementi appartenenti al gruppo delle terre rare (o serie dei lantanidi) e dei rapporti isotopici degli elementi pesanti quali lo Stronzio. La determinazione analitica delle terre rare nel pomodoro è stata effettuata con ICP-MS e HR-ICP-MS (Inductivity Coupled Plasma Mass Spectrometry, Hight Resolution Inductivity Coupled Plasma Mass Spectrometry). I rapporti isotopici dello Stronzio sono stati determinati con la tecnica TIMS (Thermal-Ionization Mass Spectroscopy) Le tecniche analitiche utilizzate hanno permesso di determinare con precisione e accuratezza tutti gli elementi appartenenti al gruppo delle terre rare nei comparti della pianta di pomodoro e nel terreno. La misura dei rapporti isotopici dello stronzio ha permesso di discriminare prodotti provenienti da aree geografiche differenti. / Food traceability systems have a broader scope and aim to document the history of a product along the entire production chain from primary raw materials to the final consumable product. the present work has looked at ways of detecting rare earth and strontium isotope abundance ratios. It is fundamental to have knowledge of the REE concentrations of the soil where the plants are grown and the distribution of the REEs in the different parts of the plants: roots, stems, leaves, and berries. Tomato is one of the most widely grown vegetables in the world. The analytical procedure, using inductively coupled mass spectrometry (ICP-MS) and HR-ICP-MS proved to be suitable for the determination of REEs in plants and soil, and was validated using certified samples. Strontium isotope ratio is an important parameter allowing the discrimination of foods coming from different geographical origins. Very high precision is usually reached with Thermal Ionization Mass Spectrometer (TIMS). The analytical techniques have permitted to determine all REEs elements in compartments of tomato plants. The described method point out the measurement of the 87Sr/86Sr ratio as a very important parameter for the discrimination of tomato from different geographical origins, even if the present experimental work treating a limited number of samples have to be considered as a “first step”.

Gli isomeri del CLA (Acido Linoleico Coniugato) potrebbero avere effetti benefici sulla salute in relazione al raggiungimento di un determinato intake, ma, per quanto ci è dato sapere, non esiste letteratura in Italia riguardante l'effettiva quantità di CLA negli alimenti e al relativo intake nell'uomo. Quindi è stata condotta una quantificazione degli isomeri c9,t11 e t10,c12 negli alimenti disponibili sul mercato italiano, seguita da una stima dell'intake di questi isomeri in una coorte della popolazione italiana attraverso un diario alimentare. Lo step successivo del progetto di ricerca, dovuto al livello molto basso di CLA trovato negli alimenti, è stato di investigare i fattori genetici che influenzano il CLA nella carne, per poter aumentare la concentrazione finale di CLA per il consumatore finale. Quindi gli obiettivi del lavoro sono stati: la stima dell'effettivo intake di CLA in Italia attraverso una completa analisi della concentrazione di c9,t11 e t10,c12 nei prodotti lattiero-caseari e carnei comunemente disponibili, e una valutazione dei fattori genetici coinvolti nella sintesi di CLA nella carne. / CLA could exert some healthy properties in relation to a needed daily intake, but, at our knowledge, no literature in Italy is currently available on the effective CLA amount in foods and on the relative intake in humans. Thus a quantification of c9,t11 and t10,c12 CLA isomers in foods available in Italy large retail-scale trade was performed, followed by an estimation of the isomers intake in a cohort of the Italian population by a food diary. Background 6 The following step of the research project, due to the very low concentration of CLA found in foods, investigated genetic factors affecting CLA in meat, in order to enhance its concentration for final consumers. Thus the aims of the present work were to estimate the effective CLA intake in Italy through a complete analisys of c9,t11 and t10,c12 concentrations in dairy and meat products commonly purchasable, and an evaluation on the genetic factors involved in CLA synthesis in meat.

L’alimentazione proteica risulta di fondamentale importanza per l’ottimizzazione della produttività e per il benessere degli animali di interesse zootecnico. L’identificazione di fonti vegetali alternative e/o complementari a quelle attualmente utilizzate (soia in primis) nel razionamento degli animali si rende necessaria per motivi economici, ambientali e legislativi, cercando di evitare, tuttavia, penalizzazioni sull’appetibilità della razione e sulla risposta produttiva. La ricerca condotta ha riguardato lo studio della composizione nutrizionale di semi di cece (Cicer arietinum) e lupino azzurro (Lupinus angustifolius), con approfondimenti sul contenuto di composti nutrizionalmente attivi e/o antinutrizionali, la valutazione della fermentescibilità ruminale mediante tecniche in vitro, anche alla luce di possibili interazioni genetiche (varietà e linee genetiche) e colturali (epoca e densità di semina). I risultati ottenuti hanno permesso di comprendere eventuali limiti di utilizzo di tali leguminose per l’alimentazione degli animali, in particolar modo dei ruminanti, e di attribuire le fonti di variabilità per la massimizzazione produttiva e qualitativa. / Ruminants protein nutrition is an essential item for optimizing the productivity and welfare of animals. The identification of pulses sources as alternative or complementary to those currently used (primarily soya) in the ration of ruminants is necessary for economic, environmental and legislative reasons, trying to avoid, however, penalties on palatability and productive response. The research focused on the study of the nutritional composition of seeds of chickpea (Cicer arietinum) and blue lupine (Lupinus angustifolius), with insights on the content of nutritionally active compounds and / or antinutritionals, the evaluation of the ruminal fermentability using in vitro techniques, also considering possible genetic interactions (varieties and genetic lines) and farming practices (time and density of sowing). The results of the present study suggest that these pulses could be a potentially good feedstuff contributing at the same time to the energy and protein balance of animal diet. Moreover, the presence of active compounds like tannins at levels that should not be regarded as anti-nutritional, can be useful in ruminant feeding due to the moderating activities exerted by those compounds toward the ruminal degradation of proteins.

La presente tesi descrive i risultati del programma di controllo dei pesticidi in regione Lombardia da 1996 a 2008 ed analizza i dati per calcolarne, con metodi diversi, la valutazione dell’esposizione del consumatore. 9387 campioni sono stati analizzati con un numero di campioni irregolari pari all’1%. Il numero di campioni senza residuo era pari al 69% ed il numero di campioni con i residui al di sotto del valore limite stabilito per legge era del 30%. Successivamente per capire l'esposizione dei consumatori a residui di antiparassitari si è utilizzato un metodo deterministico sviluppato da EFSA (PRIMo). È stato trovato che fra i campioni irregolari analizzati, solo 31 potrebbero causare il danno alla salute del consumatore. Un’ ulteriore analisi è stata quella di effettuare una valutazione con metodo probabilistico (Creme) calcolando l'esposizione cumulativa di antiparassitari sulla salute dei consumatori. Coem primo passo, residui di uno stesso pesticida trovato su campioni di patate sono stati inseriti nel software. Inoltre, campioni contenenti residui di pesticidi organofosfati sono stati inseriti nel software accoppiati con i dati italiani di consumo. In entrambi i casi, la valutazione cumulativa probabilistica dimostrava un adeguato livello di sicurezza per adulti e bambini. / The presented thesis describes the results of the pesticide monitoring programme in Lombardy Region from 1996 to 2008 and analyses the data gathered to calculate consumer exposure assessment with different approaches. A total of 9387 samples were analysed and the number of irregular samples was equal to 1%. The number of samples without residues was 69% and the number of samples with residues within the MRL was 30%. A further step to understand the exposure of consumers to residue of pesticides was obtained with the use of a deterministic approach developed by EFSA (PRIMo Model). It was found that among the detected irregular samples, only 31 might cause harm to the health of the consumer. An additional step was constituted by the use of one probabilistic method (Creme Software) to calculate the cumulative exposure of pesticides for the consumers. As a first step, residues of Chlorprofam were plotted in the software on samples of potato. In addition, samples containing residues of Organophosphates were also plotted along with the Italian consumption data. In both the case studies, the probabilistic acute cumulative assessment indicated that the intake, for adults and toddlers was below the set toxicological endpoint.

Lo scopo della tesi è di caratterizzare i rischi per la salute umana, che derivano dall’inquinamento delle acque (acque sotterranee) e dal cibo (riso) tramite l’applicazione e lo sviluppo di nuove tecniche analitiche. La tecnica “diffusive gradients in thin films (DGT)” con resina a base di ferrihydrite, è stata studiata per l'accumulo delle forme inorganiche (As(III) e As(V)) e organiche (DMA(V) e MMA(V)) di Arsenico in matrici acquose. I DGT sono stati applicati su campioni di acque sotterranee, raccolti in sei diverse città del Nord Italia, dove la concentrazione di As è molto alta. Antimonio (Sb) è associato all’As in diversi studi perché le proprietà fisiche e chimiche di questi due elementi sono simili, ed esso è stato recentemente riconosciuto come contaminante dell'acqua. In questa tesi per la prima volta sono state riportate le prestazioni dettagliate dei DGT con ferrihydrite, impiegati in soluzioni acquose di Sb(III) e Sb(V). L’analisi delle forme chimiche di Sb(III) e Sb(V) in campioni acquosi è stata realizzata con l’applicazione della diluizione isotopica. In generale il riso, a differenza di altri prodotti alimentari di origine terrestre, contiene quantità significative di Arsenico inorganico. L’analisi della speciazione di As è stata realizzata in 70 campioni di riso italiano, rappresentativo di diversi tipi di coltivazione. Le forme più abbondanti nel riso erano As(III) e DMA(V). Inoltre è stato interessante studiare la localizzazione dell’As nel chicco di riso, sottoposto a diversi processi di lavorazione (risone, riso integrale e lavorato con o senza parboiling), perché sia la distribuzione che la presenza delle diverse forme chimiche nel chicco sono fattori chiave che controllano la biodisponibilità del contaminante. La distribuzione dell’ As nei chicchi di riso di due varietà (Gladio e Ronaldo) sottoposti a diversi processi, è stato determinato con Laser Ablation ICP-MS. In realtà il contenuto di As era più alto nei chicchi di riso non-parboiled rispetto a quello parboiled. La relazione tra intensità dell’As e le varie parti del chicco di riso ha rivelato che i livelli di As diminuisco dalla parte esterna verso la posizione centrale, mentre i valori di As sembrano essere simili tra la parte media e interna dei non parboiled. / The aim of this thesis was to explore new analytical techniques as well as to carry out further characterisations of human health risks, which derive from water pollution, in particular groundwater, and food, in particular rice. The prevention of water pollution is an environmental aspect, that includes monitoring of both natural enrichment and outside pollution with routine analysis but also with new techniques, e.g. the application of passive sampling techniques and advanced technologies. The diffusive gradients in thin films (DGT) technique with ferrihydrite adsorbent, has been investigated for the accumulation of different species of Arsenic (As), like Inorganic Species (arsenite and arsenate) and Organic Arsenic (dimethylarsinic and monomethylarsenate) in aqueous matrix. To evaluate the performance of DGT method for accumulation of arsenic species, after deployment in synthetic solutions, DGT devices were carried out on groundwaters collected in six different towns in the North of Italy, where the As concentration is very high. Recently, health effects at arsenic exposures have been observed in areas where levels of inorganic As in drinking water are not excessive. Antimony (Sb) is associated to As in several studies because the physical and chemical properties of these two elements are similar, and it has been recently recognized as water contaminant. In this thesis I reported for the first time detailed performance characteristics of the Fe-oxide gel associated to DGT devices deployed in known aqueous solutions of trivalent and pentavalent Sb. Speciation analysis of Sb(III) and Sb(V) in aqueous samples was performed through extraction and on-line determination of isotope dilution concentration after a chromatographic separation. Generally rice, unlike food products of terrestrial origin, contains significant amounts of inorganic arsenic. Recently some Government Organizations (e.g. EFSA) debated the possibility to set an upper limit for total and inorganic arsenic in rice. Arsenic speciation was realized in 70 Italian rice samples from different representative cultivation conditions. The most abundant forms in rice were As(III) and DMA(V). After that, it was fundamental to investigate the localization of As in rice grains in different processes (raw, brown and milled rice with or without parboiling technique), because both speciation and distribution throughout the grain are key factors controlling bioavailability of the contaminant. The As distribution in rice grains of two varieties (Gladio and Ronaldo) from different processes, was determined by LA-ICP-MS. The distribution of As varied between the various parts of the grains (exterior, medium and interior part). During parboiling, the partial boiling of food as the first step in the cooking process, arsenic might have released from the grain to the boiling water. Thus, parboiling of rice grain may reduce the magnitude of arsenic intake in human body. Actually the As content was higher in non-parboiled rice grain than in parboiled rice. The relationship between As intensities and the different parts of rice grain revealed that As levels decreased from the external part towards the middle position, and then the intensity values seem to be similar between medium and internal part in non parboiled products.

Enterococcus faecium è un componente fondamentale del microbiota di diversi alimenti fermentati quali formaggi e salumi e viene spesso isolato in alto numero in alimenti pronti al consumo. É inoltre largamente utilizzato come probiotico sia per l’uomo che per gli animali. Allo stesso tempo, però, questa specie batterica rappresenta una delle cause principali di infezioni nosocomiali quali endocarditi ed infezioni al tratto urinario. Studi recenti hanno dimostato che la specie E. faecium è costituita da due sub-popolazioni principali: la prima è denominate hospital associated (HA) clade “A” ed include la maggior parte dei ceppi responsabili di infezioni umane; la seconda è chiamata community associated (CA) clade “B”, e contiene principalmente ceppi commensali dell’uomo. Analisi più approfondite hanno rivelato un ulteriore suddivisione all’interno del clade A, nel sub-clade A1 (che raggruppa la maggioranza dei ceppi clinici) e nel sub-clade A2, associato agli animali e più sporadicamente ad infezioni umane. Nel 2012, EFSA ha redatto una linea guida per la valutazione della sicurezza di E. faecium usato come probiotico per gli animali, concludendo che i cepi appartenenti all’hospital-associated clade non devono essere utilizzati in nutrizione animale. Comunque, la distinzione tra le due sub-popolazioni è stata fatta utilizzando dati ottenuti prevalentemente da isolati umani e animali e solo un numero limitato di ceppi isolati dagli alimenti è stato considerato. Obiettivo di questa tesi di dottorato è stato quello di valutare la sicurezza di E. faecium negli alimenti fermentati, considerando ceppi isolati da formaggi artigianali e prodotti carnei e utilizzando sia tecniche di genomica che analisi fisiologiche. Nessuno dei ceppi alimentari studiati è risultato parte del clade A1, ma un ceppo isolato da un salame stagionato pronto al consumo ha rivelato diversi tratti tipici dei ceppi A1, tra cui particolari IS, transposase e geni di resistenza agli antibiotici. Questi risultati, così come altri dati, sottolineano la necessità di approfondire le conoscenze circa il ruolo dei ceppi di E. faecium isolati da alimenti come fattore di rischio per la salute umana. / Enterococcus faecium is commonly found in high numbers in ready to eat foods, being a member of the bacterial communities of a variety of fermented foods, including cheese and sausages, and is widely used as human and animal probiotic. However, this bacterial species is a leading cause of nosocomial infection, mainly endocarditis and urinary tract infections. Recent studies have demonstrated that E. faecium species consists of two very distinct clades: the hospital associated (HA) clade “A”, which includes most of the strains responsible for human infections, and the community associated (CA) clade “B”, that contains primarily human commensal isolates. Deeper analysis revealed a further split within clade A into sub-clade A1 (which groups the vast majority of clinical isolates), and sub-clade A2, associated with animals and sporadic human infections. In 2012, the European Food Safety Authority has issued a guideline for the safety assessment of E. faecium used as animal probiotics, concluding the strains belonging to the hospital-associated clade should not be used in animal nutrition. However, the differentiation of the two clades has been performed using data mainly deriving from human and animal isolates, and only a limited number of strains from the food chain were considered. Aim of this doctoral thesis was to assess the safety of E. faecium in fermented food, considering strains isolated from artisanal cheese and meat products, and using both whole genome-based techniques and physiological studies. None of the food isolates studied in this work belong to the epidemic clade A1, however a strain isolated from a ready to eat salami revealed several A1-specific traits, such as specific IS, transposases and antibiotic resistance genes. These results, as well as other data, underline the emergency of deeper understanding the role of E. faecium isolated from fermented foods as risk factor for human health.