Lipid quality and oxidative stability in food products from animal origin, as affected by breeding factor

Nowadays it is requested more investigations on alternative rearing systems that are able to improve poultry welfare and to warrant high-quality and safe meat products. This thesis work was focused on the evaluation of the oxidative stability of poultry meats, obtained with different rearing systems, diets (supplemented with bioactive compounds), and packaging conditions. The thesis work was divided into the following parts: - Evaluation of the effects of different rearing systems on the quality, fatty acid composition and oxidative stability of poultry thigh and breast meat belonging to different product categories (“rotisserie” and “cut-up” carcasses); - Evaluation of the effects of different rearing systems and packaging conditions on the shelf-life of poultry thigh meat stored at 4°C for 14 days, and the effects of feed supplementation with thymol (control diet and diet with 2 different concentration of thymol) and packaging conditions on lipid oxidation of poultry thigh meat shelf-life (stored at 4°C for 14 days). The oxidative stability of poultry meat was studied by means of the spectrophotometric determinations of peroxide value and thiobarbituric acid reactive substances. - Evaluation of anti-inflammatory effects of different flavonoids (thymol, luteolin, tangeretin, sulforaphane, polymethoxyflavones, curcumin derivates) to detect their biological activity in LPS-stimulated RAW 264.7 macrophage cells in vitro, in order to study more in depth their action mechanisms. It was evaluated the cell vitality (MTT assay), nitrite concentration and protein profile. The study was focused on the identification of potential dietary bioactive compounds in order to investigate their biological activity and possible synergic effects, and to develop new suitable strategies for long-term promotion of human health, in particular against cancer.

Use of essential oils and biocontrol cultures for the improvement of shelf-life of fresh cut products

The demand of minimally processed fruits and vegetables has increased in the last years. However, their intrinsic characteristics may favor the growth of pathogens and spoilage microbiota. The negative effects on human health reported for some traditional chemical sanitizers have justified the search for substitutes to guarantee food safety and quality. In this work we have evaluate the potential of some essential oils and their components to improve the safety and the shelf life of Lamb’s lettuce (Valerianella locusta) and apples (Golden delicious). Moreover, the effects of selected lactic acid bacteria alone or in combination with essential oils or their components, on the shelf-life and safety as well as organoleptic properties of minimally processed products, were evaluated. Since the lack of knowledge of microbial cell targets of essential oils represent one of the most important limit to the use of these molecules at industrial level, another aim of this thesis was the study of the action mechanisms of essential oils and their components. The results obtained showed the beneficial effects of the natural antimicrobials as well as the selected lactic acid bacteria on minimally processed fruit and vegetable safety and shelf-life, without detrimental effects on the quality parameters. The beneficial effects obtained by the use of the selected biocontrol agents were further increased combining them with selected natural antimicrobials. The natural antimicrobial employed induced noticeable modifications of membrane fatty acid profiles and volatile compounds produced by microbial cells during the growth. The modification of the expression in genes involved in fatty acid biosynthesis suggesting that the cytoplasmic membrane of microbial cells is one of the major cellular target of essential oils and their components. The comprehension of microbial stress response mechanisms can contribute to the scaling up of natural antimicrobials and bio-control agents at industrial level.

Milk and dairy products: evaluation of bioactive components by analytical techniques

Milk and dairy products are important source of bioactive compounds useful to satisfy the nutritional and physiological needs of any newborns of mammalian species and useful to guarantee adequate growth and development of infants as well as provide a complete nourishment of adults. Physico-chemical, nutritional and organoleptic properties of the main constituents and the “minor” components have a crucial role in the quality of milk and milk products. Although in the past decades dietary milk fat was often regarded as harmful for the human health, recent researches suggest that milk contains specific fatty acids with nutritional and physiological health benefits. For these reasons, a major attention is given to the quantity and quality of total fat intake. In the recent years, as a result of the new concept of multifunctional agriculture and the changing behaviours about diet, consumer demands in favor of high-quality, security and safety dairy products are increased. Moreover, milk proteins and milk-derived bioactive peptides are recognized to have a high nutritive value, several health-promoting functional activities and excellent technological properties. Accordingly, growing interest in the development of functional dairy products and preparation of infant formulae for babies who cannot be breast-fed, has been give in order to meet the specific consumer’s requests. This manuscript presents the main results obtained during my PhD research aimed to evaluate the main bioactive lipids and proteins in milk and dairy products using innovative analytical techniques. The experimental section of this manuscript is divided in two sections where are reported the main results obtained during my research activities on dairy products and human milks in order to characterize their bioactive compounds for functional food applications.

Possibilities for the healthy and nutritional improvement of confectionery and sweet products.

The overall objective of this PhD was to investigate the possibility to increase the nutritional value of confectionary products by the use of natural ingredients with healthy functions. The first part of the thesis focused on the possible substitution of the most characteristic component of confectionary products, i.e. refined sugar. Many natural whole sweetening alternatives are available, though not widely used; the use of molasses, the byproduct of sugar beet and cane production, still rich in healthy components as minerals and phytochemicals is hereby discussed; after having verified molasses effectiveness in oxidative stress counteraction on liver cultured cells, the higher antioxidant capacity of a sweet food prepared with molasses instead of refined sugar was confirmed. A second step of the project dealt with another main ingredient of various sweet products, namely wheat. Particularly, the exploitation of soft and durum wheat byproducts could be another sustainable strategy to improve the healthy value of confectionery. The isolation of oligosaccharides with bioactive functions form different fractions of the wheat milling stream was studied and the new ingredients were shown to have a high dietary fiber and antioxidants content. As valid alternative, product developers should consider the appealing and healthy addition of ancient grains flour to sweet baked goods. The possibility of substituting the modern whole durum wheat with the ancient Kamut® khorasan was considered, and the antioxidant and anti-inflammatory effects of these grains were evaluated and compared both in vitro and in vivo on rats. Finally, since high consumption of confectionery is a risk factor for obesity, a possible strategy for the counteraction of this disease was investigated. The ability of three bioactives in inhibiting adipocytes differentiation was investigated. In fact, theoretically, compounds able to influence adipogenesis could be used in the formulation of functional sweet products and contribute to prevent obesity.

By-products of the dairy farming as raw material for the biotechnology production of polyhydroxyalkanoates

I Poliidrossialcanoati (PHA) sono poliesteri completamente biodegradabili, prodotti da microrganismi come fonte di energia e di carbonio per la sintesi di nuovo materiale cellulare, utilizzando come substrato materie prime rinnovabili. Questi poliesteri sono considerati potenziali candidati per la sostituzione delle materie plastiche convenzionali. Tuttavia, i più alti costi di produzione dei PHA in confronto a quelli delle materie plastiche derivanti dal petrolio, rappresentano il principale ostacolo per la parziale sostituzione di questi ultimi con i biopolimeri. Gli alti costi sono principalmente dovuti all'utilizzo di colture microbiche pure (in cui sia presente un solo ceppo batterico) e substrati puri e costosi. Nell'ultimo decennio è stato sviluppato un processo di produzione a tre stadi alternativo e potenzialmente a minor costo, basato sull'utilizzo di colture microbiche miste (Mixed Microbials Culture, MMC) e una varietà di substrati organici a costo contenuto o nullo, quali alcuni rifiuti dell’industria agro-alimentare. Il presente studio si è concentrato sulla prima fase del processo di produzione dei PHA da colture miste, la fermentazione acidogenica, utilizzando siero di latte come fonte di carbonio per produrre acidi organici. In particolare questo lavoro ha avuto come obiettivo quello di studiare come diverse condizioni operative utilizzate nella fase di fermentazione acidogenica possono influenzare la concentrazione e il profilo degli acidi organici prodotti. Sono stati valutati anche gli effetti dei diversi profili degli acidi organici sulla fase di selezione della coltura microbica, in termini di capacità di stoccaggio di PHA e composizione polimerica.

Identification and dissolution behavior of the secondary uranium minerals in the corrosion products of Depleted Uranium (DU) ammunition formed in soils

In der vorliegenden Studie wurden verschiedene Techniken eingesetzt um drei Proben (4, 7, and 8) die aus denrnKorrosionsprodukten von aus dem Kosovo Krieg stammenden Munitionskugeln, bestehend aus abgereichertem Uranrn(Depleted Uranium - DU), zu untersuchen. Als erstes Verfahren wurde die Raman-Spektroskopie eingesetzt. Hierbeirnzeigte sichin den Proben, charakterisiert durch einen Doppelpeak, die Anwesenheit von Schoepitrn(UO2)8O2(OH)12(H2O)12. Der erste und zweite Peakzeigte sich im Spektralbereich von 840,3-842,5 cm-1rnbeziehungsweise 853,6-855,8 cm-1. Diese Werte stimmen mit den Literaturwerten für Raman-Peaks für Schoepitrnüberein. Des Weiteren wurde bei dieser Untersuchungsmethode Becquerelite Ca(UO2)6O4(OH)6(H2O)8 mit einemrnPeak im Bereich zwischen 829 to 836 cm-1 gefunden. Aufgrund des Fehlens des Becquerelitespektrums in derrnSpektralbibliothek wurde eine in der Natur vorkommende Variante analysiert und deren Peak bei 829 cm-1 bestimmt,rnwas mit den Ergebnissen in den Proben korrespondiert. Mittels Röntgenbeugung (X-Ray Diffraction, XRD) zeigtenrnsich in allen Proben ähnliche Spektren. Das lässt darauf schließen, dass das pulverisierte Material in allen Probenrndas gleiche ist. Hierbei zeigte sich eine sehr gute Übereinstimmung mit Schoepit und/oder meta-rnSchoepit(UO2)8O2(OH)12(H2O)10, sowie Becquerelite. Weiterhin war weder Autunit, Sabugalit noch Uranylphosphatrnanwesend, was die Ergebnisse einer anderen Studie, durchgeführt an denselben Proben, wiederlegt. DiernAnwesenheit von P, C oder Ca im Probenmaterial konnte ausgeschlossen werden. Im Falle von Calciumkann diesrnmit der Anwesenheit von Uran erklärt werden, welches aufgrund seines Atomradius bevorzugt in Becquerelite (1:6)rneingebaut wird. Die beiden Hauptpeaks für Uran lagen im Falle von U 4f 7/2 bei 382.0 eV und im Falle von U 4f 5/2rnbei 392 eV. Diese Werte mit den Literaturwerten für Schoepit und meta-Schoepitüberein. Die Ergebnissernelektronenmikroskopischen Untersuchung zeigen U, O, Ca, Ti als dominante Komponenten in allen Messungen.rnElemente wie Si, Al, Fe, S, Na, und C wurden ebenfalls detektiert; allerdings kann nicht ausgeschlossen werden,rndass diese Elemente aus dem Boden in der unmittelbaren Umgebung der Munitionsgeschosse stammen. Gold wurdernebenfalls gemessen, was aber auf die Goldarmierung in den Probenaufbereitungsbehältern zurückgeführt werdenrnkann. Die Elektronenmikroskopie zeigte außerdem einige Stellen in denen elementares Uran und Bodenmineralernsowie sekundäre Uranminerale auftraten. Die Elementübersicht zeigt einen direkten Zusammenhang zwischen U andrnCa und gleichzeitig keine Korrelation zwischen U und Si, oder Mg. Auf der anderen Seite zeigte sich aber einrnZusammenhang zwischen Si und Al da beide Konstituenten von Bodenmineralen darstellen. Eine mit Hilfe derrnElektronenstrahlmikroanalyse durchgeführte quantitative Analyse zeigte den Massenanteil von Uran bei ca. 78 - 80%,rnwas mit den 78,2% and 79,47% für Becquerelite beziehungsweise Schoepit aufgrund ihrer Summenformelrnkorrespondiert. Zusätzlich zeigt sich für Calcium ein Massenanteil von 2% was mit dem Wert in Becquerelite (2.19%)rnrecht gut übereinstimmt. Der Massenanteil von Ti lag in einigen Fällen bei 0,77%, was auf eine noch nicht korrodierternDU-Legierung zurückzuführen ist. Ein Lösungsexperiment wurde weiterhin durchgeführt, wobei eine 0,01 M NaClO4-rnLösung zum Einsatz kam in der die verbliebene Probensubstanz der Korrosionsprodukte gelöst wurde;rnNatriumperchlorate wurde hierbei genutzt um die Ionenstärke bei 0,01 zu halten. Um Verunreinigungen durchrnatmosphärisches CO2 zu vermeiden wurden die im Versuch für die drei Hauptproben genutzten 15 Probenbehälterrnmit Stickstoffgas gespült. Eine Modelkalkulation für den beschriebenen Versuchsaufbau wurde mit Visual MINTEQrnv.3.0 für die mittels vorgenannten Analysemethoden beschriebenen Mineralphasen im pH-Bereich von 6 – 10 imrnFalle von Becquerelite, und Schoepit berechnet. Die modellierten Lösungskurven wurden unter An- und Abwesenheitrnvon atmosphärischem CO2 kalkuliert. Nach dem Ende des Lösungsexperiments (Dauer ca. 6 Monate) zeigten diernKonzentrationen des gelösten Urans, gemessen mittels ICP-OES, gute Übereinstimmung mit den modelliertenrnSchoepit und Becquerelite Kurven. Auf Grund des ähnlichen Löslichkeitverhaltens war es nicht möglich zwichen denrnbeiden Mineralen zu unterscheiden. Schoepit kontrolliert im sauren Bereich die Löslichkeit des Urans, währendrnbecquerelit im basichen am wenigsten gelöst wird. Des Weiteren bleibt festzuhalten, dass ein Anteil an CO2 in diernverschlossenen Probenbehälter eingedrungen ist, was sich mit der Vorhersage der Modeldaten deckt. Die Löslichkeitrnvon Uran in der Lösung als Funktion des pH-Wertes zeigte die niedrigsten Konzentrationen im Falle einer Zunahmerndes pH-Wertes von 5 auf 7 (ungefähr 5,1 x 10-6 mol/l) und einer Zunahme des pH-Wertes auf 8 (ungefähr 1,5 x 10-6rnmol/l bei). Oberhalb dieses Bereichs resultiert jeder weitere Anstieg des pH-Wertes in einer Zunahme gelösten Uransrnin der Lösung. Der ph-Wert der Lösung wie auch deren pCO2-Wert kontrollieren hier die Menge des gelösten Urans.rnAuf der anderen Seite zeigten im Falle von Becquerelite die Ca-Konzentrationen höhere Werte als erwartet, wobeirnwahrscheinlich auf eine Vermischung der Proben mit Bodensubstanz zurückgeführt werden kann. Abschließendrnwurde, unter Berücksichtigung der oben genannten Ergebnisse, eine Fallstudie aus Basrah (Irak) diskutiert, wo inrnzwei militärischen Konflikten Uranmunition in zwei Regionen unter verschiedenen Umweltbedingungen eingesetztrnwurden.

NMR Based Foodomics to Investigate the Digestibility of Protein-Rich Food Products

Nowadays, in developed countries, the excessive food intake, in conjunction with a decreased physical activity, has led to an increase in lifestyle-related diseases, such as obesity, cardiovascular diseases, type -2 diabetes, a range of cancer types and arthritis. The socio-economic importance of such lifestyle-related diseases has encouraged countries to increase their efforts in research, and many projects have been initiated recently in research that focuses on the relationship between food and health. Thanks to these efforts and to the growing availability of technologies, the food companies are beginning to develop healthier food. The necessity of rapid and affordable methods, helping the food industries in the ingredient selection has stimulated the development of in vitro systems that simulate the physiological functions to which the food components are submitted when administrated in vivo. One of the most promising tool now available appears the in vitro digestion, which aims at predicting, in a comparative way among analogue food products, the bioaccessibility of the nutrients of interest.. The adoption of the foodomics approach has been chosen in this work to evaluate the modifications occurring during the in vitro digestion of selected protein-rich food products. The measure of the proteins breakdown was performed via NMR spectroscopy, the only techniques capable of observing, directly in the simulated gastric and duodenal fluids, the soluble oligo- and polypeptides released during the in vitro digestion process. The overall approach pioneered along this PhD work, has been discussed and promoted in a large scientific community, with specialists networked under the INFOGEST COST Action, which recently released a harmonized protocol for the in vitro digestion. NMR spectroscopy, when used in tandem with the in vitro digestion, generates a new concept, which provides an additional attribute to describe the food quality: the comparative digestibility, which measures the improvement of the nutrients bioaccessibility.

Definition of Food Safety Criteria for Bacteria Food-Borne Pathogens in Ready to Eat products

The aims of this research study is to explore the opportunity to set up Performance Objectives (POs) parameters for specific risks in RTE products to propose for food industries and food authorities. In fact, even if microbiological criteria for Salmonella and Listeria monocytogenes Ready-to-Eat (RTE) products are included in the European Regulation, these parameters are not risk based and no microbiological criteria for Bacillus cereus in RTE products is present. For these reasons the behaviour of Salmonella enterica in RTE mixed salad, the microbiological characteristics in RTE spelt salad, and the definition of POs for Bacillus cereus and Listeria monocytogenes in RTE spelt salad has been assessed. Based on the data produced can be drawn the following conclusions: 1. A rapid growth of Salmonella enterica may occurr in mixed ingredient salads, and strict temperature control during the production chain of the product is critical. 2. Spelt salad is characterized by the presence of high number of Lactic Acid Bacteria. Listeria spp. and Enterobacteriaceae, on the contrary, did not grow during the shlef life, probably due to the relevant metabolic activity of LAB. 3. The use of spelt and cheese compliant with the suggested POs might significantly reduce the incidence of foodborne intoxications due to Bacillus cereus and Listeria monocytogenes and the proportions of recalls, causing huge economic losses for food companies commercializing RTE products. 4. The approach to calculate the POs values and reported in my work can be easily adapted to different food/risk combination as well as to any changes in the formulation of the same food products. 5. The optimized sampling plans in term of number of samples to collect can be derive in order to verify the compliance to POs values selected.

Characterization of hydrothermal carbonization products from woodchip

L’idrotermocarbonizzazione è un processo che permette di convertire un’elevata quantità di materie prime solide in carbone. Ciò viene realizzato inserendo come sostanza in input, oltre alla materia prima iniziale, acqua liquida e, successivamente, riscaldando fino a 180°C, temperatura alla quale inizia la reazione esotermica ed il processo diventa di tipo stand-alone. Tale reazione presenta un tempo di reazione variabile nel range di 4÷12 h. I prodotti in uscita sono costituiti da una sostanza solida che ha le caratteristiche del carbone marrone naturale e un’acqua di processo, la quale è altamente inquinata da composti organici. In questo elaborato viene illustrata una caratterizzazione dei prodotti in uscita da un impianto di idrotermo carbonizzazione da laboratorio, il quale utilizza in input pezzi di legno tagliati grossolanamente. Inizialmente tale impianto da laboratorio viene descritto nel dettaglio, dopodiché la caratterizzazione viene effettuata attraverso DTA-TGA dei materiali in ingresso ed uscita; inoltre altre sostanze vengono così analizzate, al fine di confrontarle col char ed i pezzi di legno. Quindi si riporta anche un’analisi calorimetrica, avente l’obiettivo di determinare il calore di combustione del char ottenuto; attraverso questo valore e il calore di combustione dei pezzi di legno è stato possibile calcolare l’efficienza di ritenzione energetica del processo considerato, così come la densificazione energetica riscontrata nel materiale in uscita. In aggiunta, è stata eseguita un’analisi delle specie chimiche elementari sul char ed il legno in modo da determinare i seguenti parametri: fattori di ritenzione e fattori di ritenzione pesati sulla massa in termini di concentrazione di C, H, N e S. I risultati ottenuti da tale analisi hanno permesso di effettuare una caratterizzazione del char. Un tentativo di attivazione del char viene riportato, descrivendo la procedura di attivazione seguita e la metodologia utilizzata per valutare il buon esito o meno di tale tentativo di attivazione. La metodologia consiste di uno studio isotermo dell’adsorbimento di acido acetico sul char “attivato” attraverso una titolazione. I risultati sperimentali sono stati fittati usando le isoterme di Langmuir e Freundlich e confrontati con le capacità di adsorbimento del semplice char e di un campione di carbone attivo preso da un’azienda esterna. Infine si è considerata l’acqua di processo, infatti un’analisi fotometrica ne ha evidenziato le concentrazioni di TOC, COD, ioni nitrato e ioni fosfato. Questi valori sono stati conseguentemente confrontati con i limiti italiani e tedeschi massimi ammissibili per acque potabili, dando quindi un’idea quantitativa della contaminazione di tale acqua di processo.

Craniofacial morphology in complete unilateral cleft lip and palate patients consecutively treated with 1-stage repair of the cleft

OBJECTIVE: To retrospectively evaluate the craniofacial morphology of children with a complete unilateral cleft lip and palate treated with a 1-stage simultaneous cleft repair performed in the first year of life. METHODS: Cephalograms and extraoral profile photographs of 61 consecutively treated patients (42 boys, 19 girls) who had been operated on at 9.2 (SD, 2.0) months by a single experienced surgeon were analyzed at 11.4 (SD, 1.5) years. The noncleft control group comprised 81 children (43 boys and 38 girls) of the same ethnicity at the age of 10.4 (SD, 0.5) years. RESULTS: In children with cleft, the maxilla and mandible were retrusive; the palatal and mandibular planes were more open, and sagittal maxillomandibular relationship was less favorable in comparison to noncleft control subjects. Soft tissues in patients with cleft reflected retrusive morphology of hard tissues--subnasal and supramental regions were less convex, profile was flatter, and nasolabial angle was more acute relative to those of the control subjects. CONCLUSIONS: Craniofacial morphology after 1-stage repair was deviated in comparison with noncleft control subjects. However, the degree of deviation was comparable with that found after treatment with alternative surgical protocols.

Comparison of two scoring systems for evaluation of treatment outcome in patients with complete bilateral cleft lip and palate

Objective : To compare two scoring systems: the Huddart/Bodenham system (HB system) and the Bauru-BCLP yardstick (BCLP yardstick), which classify treatment outcome in terms of dental arch relationships in patients with complete bilateral cleft lip and palate (CBCLP). The predictive value of these scoring systems for treatment outcome was also evaluated. Design : Retrospective longitudinal study. Patients : Dental arch relationships of 43 CBCLP patients were evaluated at 6, 9, and 12 years. Setting : Treatment outcome in BCLP patients using two scoring systems. Main Outcome Measures : For each age group, the HB scores were correlated with the BCLP yardstick scores using Spearman's correlation coefficient. The predictive value of the two scoring systems was evaluated by backward regression analysis. Results : Intraobserver Kappa values for the BCLP yardstick scoring for the two observers were .506 and .627, respectively, and the interobserver reliability ranged from .427 and .581. The intraobserver reliability for the HB system ranged from .92 to .97 and the interobserver reliability from .88 to .96. The BCLP yardstick scores of 6 and 9 years together were predictors for the outcome at 12 years (explained variance 41.3%). Adding the incisor and lateral HB scores in the regression model increased the explained variance to 67%. Conclusions : The BCLP yardstick and the HB system are reliable scoring systems for evaluation of dental arch relationships of CBCLP patients. The HB system categorizes treatment outcome into similar categories as the BCLP yardstick. In case a more sensitive measure of treatment outcome is needed, selectively both scoring systems should be used.