Il marciume del grappolo, causato da Botrytis cinerea, è una delle principali malattie che colpiscono la vite. Botrytis cinerea si sviluppa e cresce attivamente come patogeno necrotrofo e saprofita su diversi substrati. In vigneto, B. cinerea può produrre un ampio numero di conidi in un’ampia gamma di condizioni ambientali. Botrytis cinerea ha diversi pathway d’infezione e le infezioni occorrono principalmente in due periodi: dalla fioritura allo sviluppo dei giovani grappoli, e dopo invaiatura alla raccolta. A causa della complessità del ciclo di vita di B. cinerea e all’esistenza di questi diversi pathway d’infezione, i fungicidi vengono normalmente applicati a fine della fioritura (A), pre-chiusura del grappolo (B), invaiatura (C), e prima della raccolta (D). Le applicazioni durante la prima parte della stagione (A e B) hanno come obiettivo: i) la riduzione della germinazione dei conidi e le infezioni fiorali, ii) la prevenzione delle infezioni latenti delle bacche, e iii) la disinfezione dei residui di fioritura. Le applicazioni successive all’invaiatura (C e D) hanno lo scopo non solo di prevenire le infezioni durante la maturazione ma anche di disinfestare i residui di fioritura rimasti all’interno del grappolo. Questa strategia, basata su quattro trattamenti, garantisce una buona protezione contro il marciume del grappolo in vigneto, tuttavia presenta alcuni limiti: i) si tratta di un approccio preventivo che non considera il reale rischio d’infezione, con il conseguente effetto di eseguire trattamenti non necessari; ii) aumenta la probabilità che si sviluppino popolazioni resistenti ai fungicidi; iii) implica potenziali risvolti negativi sulla salute umana e sull’ambiente. Negli ultimi anni, nuove tipologie di prodotti, come gli agenti di biocontrollo (BCAs) e i botanicals, sono state proposte per ridurre l’uso eccessivo di prodotti chimici e aumentare la sostenibilità del controllo del marciume del grappolo in vigneto. Lo scopo di questa tesi di Dottorato è stato: i) comparare diverse strategie di controllo e valutare l’interazione tra fungicidi con diversi timing di applicazione, ii) valutare l’effetto sulla colonizzazione e sporulazione di B. cinerea su residui fiorali di diversi prodotti (fungicidi, agenti di biocontrollo, e botanicals) applicati con diversi timing; iii) valutare un modello meccanicistico sviluppato da González-Domínguez et al. (2015); e iv) includere all’interno di un modello per il biocontrollo, sviluppato da Jeger et al. (2009), l’effetto delle condizioni ambientali sulle interazioni BCA-patogeno. / Botrytis bunch rot (BBR), caused by Botrytis cinerea, is one of the main diseases affecting grapevine. Botrytis cinerea develops and actively grows as necrotrophic pathogen and as saprophyte on different substrates. In grapevine, B. cinerea can produce a large number of conidia on bunch and leaf trash and rotted berries under a wide range of environmental conditions. Botrytis cinerea has multiple infection pathways and infection mainly occurs in two periods: from flowering to young cluster development, and after veraison to harvest. Due to the complexity of the B. cinerea life cycle and the existence of these different infection pathways, fungicides are usually applied at the end of flowering (A), pre-bunch closure (B), veraison (C), and before harvest (D). The early season applications (A and B) are aimed at: i) reducing conidial germination and infection of flowers, ii) preventing latent infections of berries, and iii) disinfesting the bunch trash. The later-season applications (i.e., the applications from veraison until harvest; C and D) are aimed not only at preventing berry infection during ripening but also at disinfesting bunch trash in order to reduce the inoculum load. This fungicide schedule may results in unnecessary sprays because the applications are preventive and do not take into account the real risk of BBR infections. In the last decades, new products, like biological control agents (BCAs) and botanicals, have been proposed for reducing the excessive use of chemicals in agriculture and increase the sustainable of BBR control in vineyards. The aim of this Doctoral work was to: i) compare different control strategies and evaluate the interactions among fungicides applied at different timings; ii) investigate the effects of different products (fungicides, biological control agents, and botanicals) applied at different timings on B. cinerea bunch trash colonization and sporulation; iii) evaluate a weather-driven mechanistic model developed by González-Domínguez et al. (2015); and iv) include into a model for biological control, developed by Jeger et al. (2009), the effect of environmental conditions on the pathogen-BCA interactions.

In immunodeficient patients, for example with cystic fibrosis (CF), the opportunistic pathogen Staphylococcus aureus causes chronic infections of respiratory tract that are treated with antibiotics (ATB) in the long term. However, exposure to antibiotics can lead to persistence, thereby result a recurrence of infection. The aim of this work was to examine in selected pairs of S. aureus chronological isolates from the respiratory tract of CF patients how their ability to form persisters is changing in time. I have found that the ability to persist within the clonal pair does not change significantly after two years of survival in the host, and that the ability to persist depends on the adaptative mutations of the isolates. Persister formation may depend on mutations in operon of the alternative sigma B factor (sigB) and the major virulence gene regulator (agr). By dual staining with DioC2(3) and To-pro-3, I was able to determine the changes in membrane potential and membrane permeability during the killing curve with ATBs. The distribution into subpopulations according to these parameters depends primarily on the antibiotic used. I conclude that various antibiotics can induce different mechanisms causing a persistent state. Futhermore, I have constructed plasmids with a labeled promoter to determine...

La torbidità proteica nei vini bianchi e la sostanza colorante instabile nei vini rossi sono due dei principali fenomeni di instabilità della vinificazione. I coadiuvanti tradizionali hanno un elevato impatto economico e ambientale e i vini possono essere meno accettati dai consumatori. Lo scopo del lavoro è studiare nuovi coadiuvanti sostenibili per risolvere questi problemi. Per prevenire la torbidità causata dalle proteine, i produttori trattano il vino con bentonite che è un efficace agente chiarificante, ma la sua applicazione ha mostrato inconvenienti che costano 1 miliardo $ all’anno nel mondo. Un’alternativa può essere l'utilizzo di vinaccioli che sono una risorsa rinnovabile e prontamente disponibile per le cantine. Inoltre, la diffusione dell'encefalopatia spongiforme bovina ha portato a una revisione delle pratiche di produzione alimentare. Molti degli agenti chiarificanti utilizzati nella vinificazione sono di origine animale (gelatina suina, albumina d'uovo e caseina). Questi problemi hanno determinato una crescente domanda di vini ottenuti senza prodotti di origine animale. Un'alternativa può essere rappresentata dalle proteine vegetali estratte da patata e pisello. I risultati hanno mostrato le capacità chiarificanti di questi prodotti e hanno portato a identificare i migliori in base alla rimozione di proteine e polifenoli e alla valutazione sensoriale dei diversi vini trattati. / Protein turbidity in white wines and unstable coloring matter in red wines are two of the main phenomena of instability in winemaking. Traditional adjuvants involved have both high economic and environmental impact and wines can be less acceptable by consumers. The aim of this work was to investigate new and sustainable adjuvants to solve these problems. To prevent protein turbidity, winemakers add to wine bentonite that is an effective fining agent, but its application has shown drawbacks that cost $1 billion worldwide per year. A possible alternative can be the use of grape seeds that are a renewable resource and readily available to wineries during vintage. Moreover, the spread of Bovine Spongiform Encephalopathy led to a review of food production practices. Several of the fining agents used in winemaking are of animal origin (porcine gelatin, egg albumin and casein). These issues resulted in an increasing demand for wines obtained without fining agents from animal sources. An alternative can be represented by vegetal proteins extracted from potato and pea. Results shown the clarifying capacities of these agents and led to identify the best ones based on proteins and phenolics removal and sensory evaluation of different treated wines.

Staphylococcus aureus is a opportunistic pathogen that can cause severe and chronic infections. The reason of the infections relapse is often the persistence. It is about adapting to stressful conditions by inducing a dormant state, which would allow bacteria to survive exposure to antibiotics and grow again after their elimination. Bacteria that persist in the patient acquire various adaptive mutations, which are transmited creating subpopulations that have a better ability to persist. The aim of this diploma thesis was to compare individual methods of persistent study that could be used in clinical practice in the future, and at the same time to try a closer molecular characterization of the persistent state with using methods for calculating gene expression. I had chronological isolates of Staphylococcus aureus at my disposal, the initial one being the primoisolate, an isolate taken at the diagnostics of cystic fibrosis before the start of antibiotic treatment. Another was taken at a distance of three-quarters of a year and the last with a half-year interval from the previous one. Following whole genome sequencing, genes in which adaptive mutations occurred were identified. The first method determines the degree of persistence by calculating CFU (Colony Forming Units) after antibiotic treatment....

In immunodeficient patients, for example with cystic fibrosis (CF), the opportunistic pathogen Staphylococcus aureus causes chronic infections of respiratory tract that are treated with antibiotics (ATB) in the long term. However, exposure to antibiotics can lead to persistence, thereby result a recurrence of infection. The aim of this work was to examine in selected pairs of S. aureus chronological isolates from the respiratory tract of CF patients how their ability to form persisters is changing in time. I have found that the ability to persist within the clonal pair does not change significantly after two years of survival in the host, and that the ability to persist depends on the adaptative mutations of the isolates. Persister formation may depend on mutations in operon of the alternative sigma B factor (sigB) and the major virulence gene regulator (agr). By dual staining with DioC2(3) and To-pro-3, I was able to determine the changes in membrane potential and membrane permeability during the killing curve with ATBs. The distribution into subpopulations according to these parameters depends primarily on the antibiotic used. I conclude that various antibiotics can induce different mechanisms causing a persistent state. Futhermore, I have constructed plasmids with a labeled promoter to determine...

In food manufacturing, the quality control procedure is a critical activity that consists in organizing, measuring, tracking, and filing the conditions of the production process and the final product, with the goal of guaranteeing the designed quality standard. During the last 30 years, due to a mounting concern by both consumers and lawmakers, the definition of quality and the application of quality control improved drastically, and new methodologies have been developed to ensure better control of food production and to understand the effect of raw materials and the process condition on the final quality of the food product. This thesis discusses the approaches to quality control procedures in food manufacture, focusing on the relationship between the conditions of the process and the quality profile of the final product, testing in a real-case scenario of a complex production process advanced data analysis procedures. The statistical and analytical procedures proposed have been applied in a real case studio from Trentingrana cheese production, a dairy consortium in the northeast region of Italy producing a ripened semi-artisanal hard cheese under the Protected Denomination of Origin (PDO) of Grana Padano. The aim is developing tailored statistical procedures that infer the effect of the critical factors of production on quality properties of this PDO product considering its semi-artisanal production process and the presence of multiple confounding factors. The statistical analyses were applied to a dataset of measurements of physical, sensory, and chemical properties collected on cheese wheels sampled systematically to represent the variability of the production of the Trentingrana wheels over two years of production. In the first introductory chapter, after a review of the different definitions of quality, the most important quality parameters for a food product and the standard measurement techniques adopted in quality control are presented. Then, in the chapter 2, the standard procedures of data analysis are reviewed, as well as the new approaches derived from the context of the foodomic sciences and machine learning models for the analysis of quality control data in food manufacturing. Two implemented and tested practical statistical procedures in the context of the Trentingrana consortium are reported: the results are discussed according to the objectives of the quality control process, the type of data, and the organization of food production. In the first case, reported in chapter 3, Linear Mixed Model ANOVA Simultaneous Component Analysis (LMM-ASCA) was developed to investigate the effect of the dairy factory, the bimester of production, and the variability within a cheese wheel using colorimetric and textural measurements. In the second case, reported in chapter 4, a standard ASCA model with the addition of a blocking factor to include systematic error was developed to investigate the relationship between the dairy factory and bimester of production and the volatile organic compounds (VOCs) profile of Trentingrana cheese wheels. In addition, in chapter 5, an approach to relate physical measurements on Trentingrana samples with sensory evaluations of texture by a trained panel is presented. The objective of this procedure is to incorporate the quality control procedure information from different quality parameters. The development of the Partial Least Squares (PLS) predictive model, its validation, and the evaluation of its performances are discussed. In the last section (chapter 6), the development of an image analysis procedure to measure the visual quality of the rind thickness of cheese wheels is reported, comparing the performances of two different algorithms. The data analysis tools proposed in this thesis have been proved to be useful for exploring, inferring, and plotting the process quality properties and suitable for analyzing complex and unbalanced experimental designs. Furthermore, the data analysis procedures proposed improve quality control activity both at the process level and at the product level, increasing the information that is possible to extract from the measurement collected in a context where standard statistical approaches cannot infer significant information.

The mite ectoparasite Varroa destructor, poses a serious threat for the survival of the Apis mellifera colonies. The intensive use of acaricidal products is one of the most common methods for defending bees from Varroa that can cause the contamination of the wax foundation used in beekeeping. The natural honeycomb management could provide a solution for this problem, because it involves the use of frames without wax foundation which allows the bees to build a complete comb ex novo. On the other hand, colonies which are free to build cells of their choice, usually build a number of drone cells higher than colonies managed with the wax foundation. This could potentially lead to several negative consequences because the V. destructor reproductive success is greater on drone broods than on worker broods. The aim of the present study was to examine the colony development, to evaluate the honey production and to monitor the growth of V. destructor infestations and associated virus infections in Apis mellifera colonies managed by using natural honeycombs compared with the conventional management. Several colony parameters were measured in spring and summer. The strength of the colony was used to estimate the worker and drone populations. In order to measure the V. destructor infestations were used several methods, such as the natural mite fall, the powdered sugar roll, the soapy water and the brood cell uncapping. Molecular analysis was performed in order to measure the viral load of five Apis mellifera viruses. The honey produced was measured by collecting the honey stored in the supers, which are boxes placed on a beehive for bees to store. The results showed that the higher presence of drone brood in the colonies managed using the natural honeycomb did not negatively affect the colony development nor the mite V. destructor population compared to control colonies. The molecular analysis showed that the DWV was the most common virus found in bee samples, and its viral load was more influenced from the mite infestation rate than from the treatment. The analysis carried out in this study showed that the natural honeycomb management can represent a valid alternative to the wax foundation. This kind of colony management thus appears to contradict our primary hypothesis which was that letting the bees build their own honeycomb would have led to a significant increase in the V. destructor infestation. Productivity data did not provide reliable results about the difference between the natural honeycomb and the conventional colony management due to climatic adversities. Further studies will be performed to better investigate this aspect. Data about the natural mite fall and the estimation of the mite population in the phoretic/reproductive phases provided a useful starting point for further studies on the correct timing to carry out acaricide treatments both in conventional and natural honeycomb managed colonies.

Over recent decades, forest land cover is dramatically changing in European mountains and in the Alps in particular. Since the 1950s the progressive urbanization of the valleys and the abandonment of mountain and rural dwellers has intensified. More than 60% of the Trentino land, is covered by forest and mainly by high forest. This human migration have brought to a progressive shrinking of meadows and pastures due to the natural forest expansion causing a dramatic change in the landscape, the consequences of which affect biodiversity, social and cultural dynamics and landscape perception as well as ecosystem services. The objective of this research focused on the application and experimentation of advanced GIS and modeling techniques to compare aerial imagery, historical maps and data and remote sensed images to understand the past landscape changes and their dynamics in Trentino and to build future scenarios based on long-term set ofobservations. The research produced a fine scale dataset representing past forest landscape for the Trentino territory. The analysis of these output data revealed a progressive afforestation process which interested homogeneously all the Trentino territory. A future forest landscape scenarios at a detailed scale (10 m) was as well produced, to simulate the future of the forest in a protected area of Trentino, to outline if the afforestation process will continue. Along with these main output of the research, new tools for image processing and evaluation of forest changes were developed.

The brown marmorated stink bug (BMSB), Halyomorpha halys (Stål) (Hemiptera: Pentatomidae), is an invasive phytophagous species native to eastern Asia. Due to its wide host plant range, it is considered among the most harmful agricultural pests in the invaded areas, including Europe, North and South America. In addition to its impact on crops, this insect causes a nuisance in urban areas, by invading buildings for overwintering using their aggregation pheromones and emitting an unpleasant smell when disturbed. The current management of this species is mainly based on the use of chemical insecticides, which show efficacy when applied frequently, with consequent negative impact on beneficial insects in the agroecosystems. As a result, there is a desire for more environmentally friendly solutions for the management of BMSB. A valid alternative is the use of exotic and coevolved biocontrol agents, for instance, the Samurai wasp, Trissolcus japonicus (Ashmead) (Hymenoptera: Scelionidae), which is already showing promising medium and long-term results against BMSB in classical biocontrol projects. Within the frame of the least-toxic alternatives, the use of the Sterile Insect Technique (SIT) in an Area-Wide management approach is potentially a valid strategy to be included in integrated control and eradication programs. The main objective of this research, therefore, was to develop a SIT for potential use in the eradication of BMSB in newly introduced areas and/or as an additional tool to reduce its impact in areas where invasive populations have been already established. However, the effective use of SIT requires producing a high number of insects, and mass rearing for such a species is considered too complicated and expensive. To overcome this limitation, we evaluated a harvest, irradiation, and release strategy using wild overwintering adults. New live traps baited with high-load pheromones have been developed for mass trapping, exploiting the natural phenomenon ruled behaviour of aggregation of BMSB in preparation for overwintering. The captures of the new pheromone traps were compared with those of identically baited sticky panels through daily services lasting 2 weeks. Following winter diapause, the collected males were irradiated with high-energy photons at four different dosages: 16, 24, 32 and 40 Gy. The effects of irradiation on the biology i.e., fertility, longevity, and mating behaviour of overwintering BMSB males were evaluated with respect to unirradiated males. To encourage the use of SIT, a linear accelerator from a hospital facility was used for irradiation instead of the classical method involving radioactive materials. A second control 0 Gy was adopted to evaluate the possible negative effects on the biology (longevity, fertility, and fecundity) of insects due to the preparation for the irradiation. Moreover, as part of an integrated pest management (IPM) strategy, the combined use of the egg parasitoid T. japonicus and SIT was considered. The suitability of sterile eggs as oviposition substrate for T. japonicus was evaluated by comparing irradiated BMSB eggs at 40 Gy, eggs obtained by fertile female mated with sterile males irradiated at 50 Gy, and refrigerated sterile eggs currently used for laboratory rearing and field monitoring. The results of the research are encouraging for SIT applications in IPM approach. 1) The new traps caught up to 15-times more adult BMSB than the sticky panels in the two weeks of collections. 2) The developed irradiation protocol that involved a 6 MV medical linear accelerator has allowed the achievement of a high level of sterility of overwintering males (over 95%) already at 32 Gy without compromising males’ longevity and mating behaviour (in no-choice conditions). No adverse effects due to the irradiation preparation protocol were observed. 3) Sterile eggs obtained by mating sterile irradiated males and fertile females (SIT eggs), showed a higher parasitoid emergence rate (%) than both those directly irradiated and refrigerated. The difference in the emergence rate is even more evident when the eggs are 20 days old, with a parasitoid emergence rate of (75.56 %) for SIT eggs compared to those irradiated (44.68 %) and refrigerated (37.57%). In conclusion: 1) the new live traps are effective in mass trapping, 2) wild harvested males of BMSB can be used in SIT, 3) the use of linear accelerators for irradiation of BMSB is a feasible alternative to classical radioactive sources and 4) sterile eggs obtained by mating sterile males and fertile females are a valid substrate for T. japonicus oviposition, supporting the potential use of SIT on BMSB.

Combining foods, combining tools. To date, most of the published works that have attempted to relate aroma release and sensory perception are far from mimicking the real consumption context, as assessments often do not consider food oral processing during mastication and are performed on single or model foods. A multidisciplinary approach integrating in vivo aroma release, oral processing behaviour, and dynamic sensory methods is needed to have a better understanding of how the release, evolution, and fading of the flavour of real complex food matrices are being perceived during consumption. The research described in this thesis aims to investigate the relative contributions that food structure and its composition and oral processing behaviour have on aroma release and perception of food matrices made up of different components and contrasting textures, so-called composite foods. Results showed that the processes of aroma release and perception of composite foods are multidimensional phenomena and seem to be modulated by cross-modal texture-aroma interactions. While aroma release increased when a food was assessed in combination with other foods, consumer’s ability to discriminate between foods was reduced and off-flavours and texture deficiencies were masked. Furthermore, the role of oral processing behaviour on the breakdown of the food impacting aroma release and perception was highlighted. The use of a multidisciplinary approach provided more useful and accurate sensory profile representing an ecological, valid food consumption context that will ultimately offer new strategies for successfully developing healthier food products with lower fat and sugar content or elaborated with plant protein, for which sensory acceptability remains a challenge.