The role of bifidobacteria in newborn health and the intestinal microbial balance

Gut microbial acquisition during the early stage of life is an extremely important event since it affects the health status of the host. In this contest the healthy properties of the genus Bifidobacterium have a central function in newborns. The aim of this thesis was to explore the dynamics of the gut microbial colonization in newborns and to suggest possible strategies to maintain or restore a correct balance of gut bacterial population in infants. The first step of this work was to review the most recent studies on the use of probiotics and prebiotics in infants. Secondly, in order to prevent or treat intestinal disorders that may affect newborns, the capability of selected Bifidobacterium strains to reduce the amount of Enterobacteriaceae and against the infant pathogen Streptococcus agalactiae was evaluated in vitro. Furthermore, the ability of several commercial fibers to stimulate selectively the growth of bifidobacterial strains was checked. Finally, the gut microbial composition in the early stage of life in response to the intrapartum antibiotic prophylaxis (IAP) against group B Streptococcus was studied using q-PCR, DGGE and next generation sequencing. The results globally showed that Bifidobacterium breve B632 strain is the best candidate for the use in a synbiotic product coupled to a mixture of two selected prebiotic fibers (galactooligosaccharides and fructooligosaccharides) for gastrointestinal disorders in infants. Moreover, the early gut microbial composition was affected by IAP treatment with infants showing lower counts of Bifidobacterium spp. and Bacteroides spp. coupled to a decrement of biodiversity of bacteria, compared to control infants. These studies have shown that IAP could affect the early intestinal balance in infants and they have paved the way to the definition of new strategies alternative to antibiotic treatment to control GBS infection in pregnant women.

Sublethal effects of a common neonicotinoid pesticide, thiamethoxam, on honey bees: impact on locomotion and thermoregulation

Neonicotinoids have been pointed to as a factor responsible for the increased honey bee colony losses in the last decades. Many studies have investigated the effects of the first marketed neonicotinoid, imidacloprid, while fewer have focused on thiamethoxam. One recent study showed that sublethal doses of thiamethoxam lead to colony failure by decreasing forager homing flight success. We thus decided to investigate the mechanism which caused this phenomenon. Our hypothesis was that this effect was caused by impairment of forager locomotion abilities. Therefore we tested the effects of sublethal acute and chronic exposures to thiamethoxam on forager walking (Chapter 2) and flight (Chapter 3) performances. The acute treatment (1.34 ng/bee) affected walking locomotion firstly triggering hyperactivity (30 min post-treatment) and then impairing motor functioning (60 min post-treatment). 2-day continuous exposures to thiamethoxam (32.5, 45 ppb) elicited fewer effects on walking locomotion, however both exposure modes elicited an increased positive phototaxis. Similarly, in flight experiments, the single dose (1.34 ng/bee) elicited hyperactivity shortly after intoxication (increased flight duration and distance), while longer and continuous exposures (32.5, 45 ppb) impaired forager motor functions (decreased flight duration, distance, velocity). It is known that flight muscles temperature needs to be precisely regulated by bees during flight. Therefore, we further hypothesized that the impaired flight performances of neonicotinoid intoxicated bees were caused also by thermoregulation anomalies. We tested the effects that acute thiamethoxam exposures (0.2, 1, 2 ng/bee) elicit on forager thorax temperature (Chapter 4). Foragers treated with high doses exhibited hyperthermia or hypothermia when respectively exposed to high or low environmental temperatures. In summary, we show that sublethal doses of thiamethoxam affected forager walking and flight locomotion, phototaxis and thermoregulation. We also display the intricate mode of action of thiamethoxam which triggered, at different extents, inverse sublethal effects in relation to time and dose.

Basi molecolari ed implicazioni fisio-patologiche dell'alterata glicosilazione di IgG nell'invecchiamento

Introduzione: Le catene N-linked associate al principale sito di N-glicosilazione (Asn297) delle IgG sono di tipo bi-antennario e presentano una grande microeterogeneità in quanto una o entrambe le antenne possono terminare con uno o due residui di acido sialico, galattosio o N-acetilglucosammina ed essere core-fucosilate. Nell’invecchiamento e in malattie infiammatorie aumenta la percentuale di glicani associati alle catene pesanti delle IgG privi del galattosio terminale (IgG-G0). La glicosilazione enzimatica delle proteine è classicamente un processo intracellulare, sebbene recenti studi abbiano messo in evidenza la possibilità di una glicosilazione ecto-cellulare in quanto le piastrine sono ottimi donatori di nucleotidi-zuccheri. Scopo: Misurare le attività delle glicosiltrasferasi ST6Gal1 e B4GalT plasmatiche (potenzialmente responsabili della glicosilazione di proteine plasmatiche) in soggetti di entrambi i sessi e di età compresa tra 5 e 105 anni e correlarle con lo stato di glicosilazione di IgG circolanti (analizzato mediante lectin-blot) e il GlycoAge test, un noto marcatore di invecchiamento, espresso come il logaritmo del rapporto tra gli N-glicani agalattosilati e di-galattosilati associati a glicoproteine plasmatiche. Risultati e conclusioni: I dati ottenuti indicano che: 1) l’attività B4GalT si propone come nuovo marcatore di invecchiamento perché aumenta linearmente con l’età; 2) la ST6Gal1 è maggiormente espressa solo nei bambini e negli over 80; 3) le attività delle due glicosilatransferasi non risultano correlate in modo significativo né tra loro né con il GlycoAge test, indicando che questi tre marcatori siano espressioni di diversi quadri fisio-patologici legati all’invecchiamento; 4) con l’età si ha una predominanza di glicoforme di IgG pro-infiammatorie, ovvero prive dell’acido sialico, del galattosio terminali e del core fucose; 5) l’attività della ST6Gal1 e B4GalT risultano in controtendenza con il grado di sialilazione e galattosilazione delle IgG, indicando quindi che la loro glicosilazione non avviene a livello extracellulare.

Water, sediment and soil physicochemical interactions in freshwater, brackish and saline systems

The physicochemical interactions between water, sediment and soil deeply influence the formation and development of the ecosystem. In this research, different freshwater, brackish and saline subaqueous environments of Northern Italy were chosen as study area to investigate the physicochemical processes which occur at the interface between water and sediments, as well as the effects of soil submergence on ecosystem development. In the freshwater system of the Reno river basin, the main purpose was to define the heavy metals hazard in water and sediments of natural and artificial water courses. Heavy metals partitioning and speciation allowed to assess the environmental risk linked to the critical action of dredging canal sediments, for the maintenance of the hydraulic safety of plain lands. In addition, some bioremediation techniques were experimented for protecting sediments from heavy metals contamination, and for giving an answer to the problem of sediments management. In the brackish system of S. Vitale park, the development of hydromorphic and subaqueous soils was investigated. The study of soil profiles highlighted the presence of a soil continuum among pedons subjected to different saturation degrees. This investigation allowed to the identification of both morphological and physicochemical indicators, which characterize the formation of subaqueous soils and describe the soil hydromorphism in transitional soil systems. In the saline system of Grado lagoon, an ecosystem approach was used to define the role of water oscillation in soil characterization and plants colonization. This study highlighted the close relationship and the mutual influence of soil submergence and aeration, tide oscillation and vegetation cover, on the soil development. In view of climate change, this study contribute to understand and suppose how soil and landscape could evolve. However, a complete evaluation of hydromorphic soil functionality will be achieved only involving physiological and biochemical expertise in these kind of studies.

La distribuzione del potere tra le Istituzioni Europee e la procedura di co-decisione. Il caso della riforma della Politica Agricola Comune

Questa ricerca analizza le implicazioni derivanti dall’introduzione della procedura di co-decisione come procedura legislativa ordinaria nel processo di riforma della politica agricola comune. La diversa distribuzione dei poteri tra le istituzioni europee modifica gli assetti istituzionali e fornisce al Parlamento il ruolo di colegislatore in materia agricola. La forma assunta dalla nuova politica agricola europea scaturisce dalla configurazione dei poteri di contrattazione che ciascun attore ha mostrato nella sede dei triloghi negoziali. La ricerca tenta di verificare la accuratezza predittiva di diversi modelli di contrattazione legislativa attraverso il confronto e la verifica degli errori di predizione sui risultati finali di alcune questioni salienti della riforma della Politica Agricola Comune e allo stesso tempo, cerca di identificare il peso del potere del Parlamento europeo in veste di co-legislatore nel processo di riforma della PAC post-2013.

Farming and processing factors for improving quality properties of poultry and rabbit meat

Recently, global meat market is facing several dramatic changes due to shifting in diet and life style, consumer demands, and economical considerations. Firstly, there was a tremendous increase in the poultry meat demand. Furthermore, current forecast and projection studies pointed out that the expansion of the poultry market will continue in future. In response to this demand, there was a great success to increase growth rate of meat-type chickens in the last few decades in order to optimize the production of poultry meat. Accordingly, the increase of growth rate induced the appearance of several muscle abnormalities such as pale-soft-exudative (PSE) syndrome and deep-pectoral-myopathy (DPM) and more recently white striping and wooden breast. Currently, there is growing interest in meat industry to understand how much the magnitude of the effect of these abnormalities on different quality traits for raw and processed meat. Therefore, the major part of the research activities during the PhD project was dedicated to evaluate the different implications of recent muscle abnormalities such as white striping and wooden breast on meat quality traits and their incidence under commercial conditions. Generally, our results showed that the incidence of these muscle abnormalities was very high under commercial conditions and had great adverse impact on meat quality traits. Secondly, there is growing market share of convenient, healthy, and functional processed meat products. Accordingly, the remaining part of research activities of the PhD project was dedicated to evaluate the possibility to formulate processed meat products with higher perceived healthy profile such as phosphate free-marinated chicken meat and low sodium-marinated rabbit meat products. Overall all findings showed that sodium bicarbonate can be considered as promising component to replace phosphates in meat products, while potassium chloride under certain conditions was successfully used to produce low marinated rabbit meat products.

Analysis of the pig genome for the identification of genomic regions affecting production traits

The aim of this work was to identify markers associated with production traits in the pig genome using different approaches. We focused the attention on Italian Large White pig breed using Genome Wide Association Studies (GWAS) and applying a selective genotyping approach to increase the power of the analyses. Furthermore, we searched the pig genome using Next Generation Sequencing (NSG) Ion Torrent Technology to combine selective genotyping approach and deep sequencing for SNP discovery. Other two studies were carried on with a different approach. Allele frequency changes for SNPs affecting candidate genes and at Genome Wide level were analysed to identify selection signatures driven by selection program during the last 20 years. This approach confirmed that a great number of markers may affect production traits and that they are captured by the classical selection programs. GWAS revealed 123 significant or suggestively significant SNP associated with Back Fat Thickenss and 229 associated with Average Daily Gain. 16 Copy Number Variant Regions resulted more frequent in lean or fat pigs and showed that different copies of those region could have a limited impact on fat. These often appear to be involved in food intake and behavior, beside affecting genes involved in metabolic pathways and their expression. By combining NGS sequencing with selective genotyping approach, new variants where discovered and at least 54 are worth to be analysed in association studies. The study of groups of pigs undergone to stringent selection showed that allele frequency of some loci can drastically change if they are close to traits that are interesting for selection schemes. These approaches could be, in future, integrated in genomic selection plans.

Entomological monitoring in the cultural heritage facilities as prerequisite for a successful IPM programme

The European Committee for Standardization is working on a standard for the application of IPM (Integrated Pest Management) in museums and cultural heritage facilities. Since one of the aims of this research was to verify the approach against pests adopted in Italian conservation facilities, a survey was conducted. The results show that for the Italian museums, archives, libraries and historical houses pests are a problem, but IPM is unknown and prevention programmes to avoid damages caused by them, are not applied. In the most of cases pests problems are solved only when the risk is high and damages are visible. Also entomological monitoring, which represents a crucial part of IPM and could be very useful, is not included among the ordinary prevention activities. In addition, at present, the scientific researches on entomological traps, whether light or pheromones, for “cultural heritage pests” is extremely poor and only recently the behaviour and/or the physiology of the insects “of museums” have been investigated. For these reasons, tests to increase the traps using are performed. In particular, S. paniceum behaviour towards different attraction systems was investigated and the results indicate that the light traps efficiency could be improved using specific wavelengths and light sources.

Identification of CNV and QTL for productive and functional traits in dairy cattle using dense SNP chips

The thesis identify CNV structural variants as possible markers for genomic selection and identify QTL regions for Fatty Acid Content in the Italian Brown Swiss population. Additionally it maps the QTL for mastitis resistance in the Valdostana Red Pied cattle.

Sviluppo di mezzi e sistemi di sorveglianza e monitoraggio dei ditteri culicidi di importanza sanitaria

Nel corso degli ultimi anni le problematiche legate al ruolo vettore delle zanzare stanno emergendo sia per quanto riguarda l’uomo che gli animali allevati e selvatici. Diversi arbovirus come West Nile, Chikungunya, Usutu e Dengue, possono facilmente spostarsi a livello planetario ed essere introdotti anche nei nostri territori dove possono dare avvio a episodi epidemici. Le tecniche di monitoraggio e sorveglianza dei Culicidi possono essere convenientemente utilizzate per il rilevamento precoce dell’attività virale sul territorio e per la stima del rischio di epidemie al fine dell’adozione delle opportune azioni di Sanità Pubblica. Io scopo della ricerca del dottorato è inserito nel contesto dei temi di sviluppo del Piano regionale sorveglianza delle malattie trasmesse da vettori in Emilia Romagna. La ricerca condotta è inquadrata prevalentemente sotto l’aspetto entomologico applicativo di utilizzo di dispositivi (trappole) che possano catturare efficacemente possibili insetti vettori. In particolare questa ricerca è stata mirata allo studio comparativo in campo di diversi tipi di trappole per la cattura di adulti di zanzara, cercando di interpretare i dati per capire un potenziale valore di efficacia/efficienza nel rilevamento della circolazione virale e come supporto alla pianificazione della rete di sorveglianza dal punto di vista operativo mediante dispositivi adeguati alle finalità d’indagine. Si è cercato di trovare un dispositivo idoneo, approfondendone gli aspetti operativi/funzionali, ai fini di cattura del vettore principale del West Nile Virus, cioè la zanzara comune, da affiancare all’unica tipologia di trappola usata in precedenza. Le prove saranno svolte sia in campo che presso il laboratorio di Entomologia Medica Veterinaria del Centro Agricoltura Ambiente “G. Nicoli” di Crevalcore, in collaborazione con il Dipartimento di Scienze e Tecnologie Agroambientali della Facoltà di Agraria dell’Università di Bologna.

From fossil to sustainable diets: an assessment of farming energy footprint

The dependence of industrial agricolture on fossil fuels has been assessed in two comparative case studies between Italy (Emilia-Romagna and Piemonte)and Missouri. The first is related to dairy farming; 15 different farms were surveyed, divided into three different groups: grain based, pasture based and organic. The second is devoted to rice cropping; 12 holdings were examined divided into two groups: conventional and organic. Energy footprint was determined for structures, machinery, fertilizers, pesticides, fuel, electricity, feed and seeds. Possible scenarios of transition to a more sustainable agricolture based on renewable energy sources were analized in detail for all the farms analized.

Study of the effect of minor compounds on the oxidative stability and physical properties of bulk oil, oil-in-water emulsion, and food emulsions

Minor components are of particular interest due to their antioxidant and biological properties. Various classes of lipophilic minor components (plant sterols (PS) and α-tocopherol) were selected as they are widely used in the food industry. A Fast GC-MS method for PS analysis in functional dairy products was set up. The analytical performance and significant reduction of the analysis time and consumables, demonstrated that Fast GC-MS could be suitable for the PS analysis in functional dairy products. Due to their chemical structure, PS can undergo oxidation, which could be greatly impacted by matrix nature/composition and thermal treatments. The oxidative stability of PS during microwave heating was evaluated. Two different model systems (PS alone and in combination) were heated up to 30 min at 1000 W. PS degraded faster when they were alone than in presence of TAG. The extent of PS degradation depends on both heating time and the surrounding medium, which can impact the quality and safety of the food product destined to microwave heating/cooking. Many minor lipid components are included in emulsion systems and can affect the rate of lipid oxidation. The oxidative stability of oil-in-water (O/W) emulsions containing PS esters, ω-3 FA and phenolic compounds, were evaluated after a 14-day storage at room temperature. Due to their surface active character, PS could be particularly prone to oxidation when they are incorporated in emulsions, as they are more exposed to water-soluble prooxidants. Finally, some minor lipophilic components may increase oxidative stability of food systems due to their antioxidant activity. á-tocopherol partitioning and antioxidant activity was determined in the presence of excess SDS in stripped soybean O/W emulsions. Results showed that surfactant micelles could play a key role as an antioxidant carrier, by potentially increasing the accessibility of hydrophobic antioxidant to the interface.

Entrepreneurship influential factors in development of rural tourism as diversified rural activity in Republic of Macedonia

Rural tourism is relatively new product in the process of diversification of the rural economy in Republic of Macedonia. This study used desk research and life story interviews of rural tourism entrepreneurs as qualitative research method to identify prevalent success influential factors. Further quantitative analysis was applied in order to measure the strength of influence of identified success factors. The primary data for the quantitative research was gathered using telephone questionnaire composed of 37 questions with 5-points Likert scale. The data was analyzed using Partial Least Squares Structural Equation Modeling (PLS-SEM) by SmartPLS 3.1.6. Results indicated that human capital, social capital, entrepreneurial personality and external business environment are predominant influential success factors. However, human capital has non-significant direct effect on success (p 0.493) nonetheless the effect was indirect with high level of partial mediation through entrepreneurial personality as mediator (VAF 73%). Personality of the entrepreneur, social capital and business environment have direct positive affect on entrepreneurial success (p 0.001, 0.003 and 0.045 respectably). Personality also mediates the positive effect of social capital on entrepreneurial success (VAF 28%). Opposite to the theory the data showed no interaction between social and human capital on the entrepreneurial success. This research suggests that rural tourism accommodation entrepreneurs could be more successful if there is increased support in development of social capital in form of conservation of cultural heritage and natural attractions. Priority should be finding the form to encourage and support the establishment of formal and informal associations of entrepreneurs in order to improve the conditions for management and marketing of the sector. Special support of family businesses in the early stages of the operation would have a particularly positive impact on the success of rural tourism. Local infrastructure, access to financial instruments, destination marketing and entrepreneurial personality have positive effect on success.

Linking rural tourism to agriculture. Evidences from comparative case studies in Kosovo, Italy and Spain

Rural tourism has been widely promoted in the European Union as an effective measure counteracting economic and social challenges facing rural areas especially those with declining agriculture economies. Particularly its role is seen in provision and maintenance of public goods which are more and more demanded by the public and considered in the policymaking. In Kosovo, rural tourism has been developed through the support of the international organizations and private sector initiatives, with primary aim to generate additional income for rural households and sustainable management of natural and cultural resources. Anyhow, it could be stated that the use of territorial capital to enhance the quality of the tourist offer and undertake promotion at wider circles of people has not been well explored so far, particularly possible links with agriculture that would satisfy visitors demand. In this regard this research study analyzes involvement of local stakeholders and use of territorial capital to develop tourist offer in rural areas of Kosovo. Beside, study applies comparative approach with other two areas of the European Union, Appennino Bolognese in Italy and Alpujara in Spain, to understand and compare the process of rural tourism development and demand characteristics between Kosovo and these areas. A survey has been conducted in all three study areas with rural tourism visitors to understand their preferences for public and private goods and services when visiting rural areas and the role of agriculture in sustaining rural tourism. Results show that there is a potential to link rural tourism with agriculture in Kosovo, which would help in sustaining agriculture and add additional value to local food products, which in return would enhance the tourist offer and make it more attractive for the visitors but also for the farmers as an additional revenue generating sector.

Biochar in perennial crops: nutritional, agronomical and environmental implications

Biochar is the solid C-rich matrix obtained by pyrolysis of biomasses, currently promoted as a soil amendment with the aim to offset anthropogenic C emissions, while ameliorating soil properties and growth conditions. Benefits from biochar seem promising, although scientific understandings are beginning to be explored. In this project, I performed a suite of experiments in controlled and in field conditions with the aims to investigate the effect of biochar on: a) the interaction with minerals; b) Fe nutrition in kiwifruit; c) soil leaching, soil fertility, soil CO2 emissions partitioning, soil bacterial profile and key gene expression of soil nitrification-involved bacteria; d) plant growth, nutritional status, yield, fruit quality and e) its physical-chemical changes as affected by long-term environmental exposure. Biochar released K, P and Mg but retained Fe, Mn, Cu and Zn on its surface which in turn hindered Fe nutrition of kiwifruit trees. A redox reaction on the biochar surface exposed to a Fe source was elucidated. Biochar reduced the amount of leached NH4+-N but increased that of Hg, K, P, Mo, Se and Sn. Furthermore, biochar synergistically interacted with compost increasing soil field capacity, fertility, leaching of DOC, TDN and RSOC, suggesting a priming effect. However, in field conditions, biochar did not affect yield, nutritional status and fruit quality. Actinomadura flavalba, Saccharomonospora viridis, Thermosporomyces composti and Enterobacter spp. were peculiar of the soil amended with biochar plus compost which exhibited the highest band richness and promoted gene expression levels of Nitrosomonas spp., Nitrobacter spp. and enzymatic-related activity. Environmental exposure reduced C, K, pH and water infiltration of biochar which instead resulted in a higher O, Si, N, Na, Al, Ca, Mn and Fe at%. Oxidation occurred on the aged biochar surface, it decreased progressively with depth and induced the development of O-containing functional groups, up to 75nm depth.