Food safety and zoonotic enteric pathogens: sources, risk factors and transmission routes of human salmonellosis and campylobacteriosis

Salmonella and Campylobacter are common causes of human gastroenteritis. Their epidemiology is complex and a multi-tiered approach to control is needed, taking into account the different reservoirs, pathways and risk factors. In this thesis, trends in human gastroenteritis and food-borne outbreak notifications in Italy were explored. Moreover, the improved sensitivity of two recently-implemented regional surveillance systems in Lombardy and Piedmont was evidenced, providing a basis for improving notification at the national level. Trends in human Salmonella serovars were explored: serovars Enteritidis and Infantis decreased, Typhimurium remained stable and 4,[5],12:i:-, Derby and Napoli increased, suggesting that sources of infection have changed over time. Attribution analysis identified pigs as the main source of human salmonellosis in Italy, accounting for 43–60% of infections, followed by Gallus gallus (18–34%). Attributions to pigs and Gallus gallus showed increasing and decreasing trends, respectively. Potential bias and sampling issues related to the use of non-local/non-recent multilocus sequence typing (MLST) data in Campylobacter jejuni/coli source attribution using the Asymmetric Island (AI) model were investigated. As MLST data become increasingly dissimilar with increasing geographical/temporal distance, attributions to sources not sampled close to human cases can be underestimated. A combined case-control and source attribution analysis was developed to investigate risk factors for human Campylobacter jejuni/coli infection of chicken, ruminant, environmental, pet and exotic origin in The Netherlands. Most infections (~87%) were attributed to chicken and cattle. Individuals infected from different reservoirs had different associated risk factors: chicken consumption increased the risk for chicken-attributed infections; animal contact, barbecuing, tripe consumption, and never/seldom chicken consumption increased that for ruminant-attributed infections; game consumption and attending swimming pools increased that for environment-attributed infections; and dog ownership increased that for environment- and pet-attributed infections. Person-to-person contacts around holiday periods were risk factors for infections with exotic strains, putatively introduced by returning travellers.

Legal presumptions in national tax systems (Italy and Belgium) and in EU law

The thesis deals with the concept of presumptions, and in particular of legal presumptions, in the context of national tax systems (Italy and Belgium) and EU law. The purpose was to investigate the concept of legal presumption under a twofold comparative perspective. After having provided a general overview of the common core concept of presumption in the European context, an insight in the national approach to legal presumptions was given by examining two different national experiences, namely the Italian and Belgian tax systems. At this stage, the Constitutional framework and some of the most interesting and relevant at EU level presumptive measures were explored, with a view to underlining possible divergences and common grounds. The concept of (national) legal presumption was then investigated in the context of EU law, with the attempt to systematize under a uniform perspective a matter which has been traditionally dealt with either from the merely national point of view or, at EU level, through a fragmented form. In this instance, the EU law relevant framework and the most significant EUCJ case-law, in particular in the field of customs duties, VAT, on the issue of the repayment of taxes levied in breach of EU law and in the area of direct taxation, were examined so as to construe the overall EU approach to national legal presumptions. This was done with the finality of determining if and to what extent a common analytical framework may be identified, from which were extracted certain criteria governing the compatibility of national legal presumptions with EU law.

Alle origini delle case di correzione. Indagine storico-educativa sulle pratiche di internamento in Italia e in Inghilterra

La presente tesi intende compiere un’indagine, dal punto di vista storico-educativo, sulla storia delle case di correzione. Nello specifico si è tentato di analizzare il “caso” del Discolato bolognese, operando un confronto con altre analoghe istituzioni, sorte in due città campione, come Roma e Milano, per quanto riguarda il contesto italiano, e con la situazione inglese, per quanto riguarda il contesto internazionale. Il focus della ricerca si è incentrato sul rapporto tra devianza e internamento, considerato secondo una declinazione pedagogica. A tal proposito si è cercato di far emergere le modalità educative, nonché i principi e le finalità che sottostavano agli interventi istituzionali, non solo soffermandosi su quanto i regolamenti e gli statuti prescrivevano all’interno delle strutture correzionali, ma analizzando la reclusione nella sua effettiva organizzazione quotidiana. Per quanto riguarda la situazione bolognese è stata analizzata un’ampissima documentazione, conservata presso l’Archivio storico Provinciale di Bologna, che ha permesso di svolgere un’analisi di tipo quantitativo su un totale di 1105 individui al fine di delineare le principali caratteristiche demografiche e sociali delle persone internate nel Discolato bolognese. L’interpretazione delle fonti ha permesso anche un’indagine qualitativa, nel tentativo di ricostruire le storie di vita dei singoli reclusi. Da questa ricerca emerge un’immagine complessa delle case di correzione. Nel corso dei secoli, in modo particolare nelle diverse realtà prese in esame, esse hanno assunto caratteristiche e peculiarità differenti. La loro inefficacia fu la motivazione principale che condusse alla definitiva chiusura, quando si fece via via sempre più evidente la difficoltà a tradurre in pratica ciò che regolamenti e statuti - a livello teorico - prescrivevano.

Geotechnical characterization of mixed sandy and silty soils using piezocone tests: Analysis of partial drainage phenomena and rate effects on the experimental soil response

The cone penetration test (CPT), together with its recent variation (CPTU), has become the most widely used in-situ testing technique for soil profiling and geotechnical characterization. The knowledge gained over the last decades on the interpretation procedures in sands and clays is certainly wide, whilst very few contributions can be found as regards the analysis of CPT(u) data in intermediate soils. Indeed, it is widely accepted that at the standard rate of penetration (v = 20 mm/s), drained penetration occurs in sands while undrained penetration occurs in clays. However, a problem arise when the available interpretation approaches are applied to cone measurements in silts, sandy silts, silty or clayey sands, since such intermediate geomaterials are often characterized by permeability values within the range in which partial drainage is very likely to occur. Hence, the application of the available and well-established interpretation procedures, developed for ‘standard’ clays and sands, may result in invalid estimates of soil parameters. This study aims at providing a better understanding on the interpretation of CPTU data in natural sand and silt mixtures, by taking into account two main aspects, as specified below: 1)Investigating the effect of penetration rate on piezocone measurements, with the aim of identifying drainage conditions when cone penetration is performed at a standard rate. This part of the thesis has been carried out with reference to a specific CPTU database recently collected in a liquefaction-prone area (Emilia-Romagna Region, Italy). 2)Providing a better insight into the interpretation of piezocone tests in the widely studied silty sediments of the Venetian lagoon (Italy). Research has focused on the calibration and verification of some site-specific correlations, with special reference to the estimate of compressibility parameters for the assessment of long-term settlements of the Venetian coastal defences.

A 300 million year-long history. The metamorphic evolution of the Northern Apennine Variscan basement

The Variscan basement of Northern Apennines (Northern Italy) is a polymetamorphic portion of continental crust. This thesis investigated the metamorphic history of this basement occurring in the Cerreto Pass, in the Pontremoli well, and in the Pisani Mountains. The study comprised fieldwork, petrography and microstructural analysis, determination of the bulk rock and mineral composition, thermodynamic modelling, conventional geothermobarometry, monazite chemical dating and Ar/Ar dating of muscovite. The reconstructed metamorphic evolution of the selected samples allowed to define a long-lasting metamorphic history straddling the Variscan and Alpine orogenesis. Some general petrological issues generally found in low- to medium-grade metapelites were also tackled: (i) With middle-grade micaschist it is possible to reconstruct a complete P-T-D path by combining microstructural analysis and thermodynamic modelling. Prekinematic white mica may preserve Mg-rich cores related to the pre-peak stage. Mn-poor garnet rim records the peak metamorphism. Na-rich mylonitic white mica, the XFe of chlorite and the late paragenesis may constrain the retrograde stage. (ii) Metapelites may contain coronitic microstructures of apatite + Th-silicate, allanite and epidote around unstable monazite grains. Chemistry and microstructure of Th-rich monazite relics surrounded by this coronitic microstructure may suggest that monazite mineral was inherited and underwent partial dissolution and fluid-aided replacement by REE-accessory minerals at 500-600°C and 5-7 kbar. (iii) Fish-shaped white mica is not always a (prekinematic) mica-fish. Observed at high-magnification BSE images it may consist of several white mica formed during a mylonitic stage. Hence, the asymmetric foliation boudin is a suitable microstructure to obtain geochronological information about the shearing stage. (iv) Thermodynamic modelling of a hematite-rich metasedimentary rock fails to reproduce the observed mineral compositions when the bulk Fe2O3 is neglected or determined through titration. The mismatch between observed and computed mineral compositions and assemblage is resolved by tuning the effective ferric iron content by P-XFe2O3 diagrams.

[Case report: an exceptional case of a monozygotic thoracopagus parasiticus in a German Holstein calf]

A black and white German Holstein calf displayed a complex double malformation in shape of a thoracopagus parasiticus. By means of a molecular genetic investigation the genesis of the malformation from one zygote could be demonstrated. Both vertebral columns showed a pronounced lordosis, with the vertebral column of one animal ending in a rudimentary head. Close to this rudiment two derivates of branchial arches were found. The two thoracic cavities merged into one "thorax". In the shared thoracic cavity one heart was found. In its right atrium, a cherry-sized structure was found in which heart- and vascular smooth muscles were demonstrated histologically. The aorta split shortly after its origin to provide both animals with one aorta each. The larger pair of lungs was connected with a trachea leading to the head while the smaller pair of lungs originated from a trachea deriving from the rudimentary head. The diaphragm jejunum and split afterwards. The pedigree of the affected animal showed neither inbreeding nor any other affected animal.

Urinary metabolites and antioxidant products of exogenous melatonin in the mouse

Exogenous melatonin is widely used for sleep disorders and has potential value in neuroprotection, cardioprotection and as an antioxidant. Here, a novel method is described for the determination of melatonin and six metabolites in mouse urine by use of LC-MS/MS and GC-MS. LC-MS/MS is used for the measurement of melatonin, N1-acetyl-5-methoxykynuramine (AMK), N1-acetyl-N2-formyl-5-methoxykynuramine (AFMK) and 6-hydroxymelatonin (6-HMEL), while GC/MS is used for the determination of N-[2-(5-methoxy-2-oxo-2,3-dihydro-1H-indol-3-yl)-ethyl]-acetamide (2-OMEL) and cyclic 3-hydroxymelatonin (3-HMEL) with detection limits on column of 0.02-0.5 pmol, depending on the metabolite. Following oral administration of melatonin to mice, a 0-24 hr urine collection revealed the presence of melatonin (0.2% dose), 6-HMEL (37.1%) and NAS (3.1%) comprising >90% of the total metabolites; AMK and AFMK were also detected at 0.01% each; 2-OMEL was found at 2.2% of the dose, which is >100 times more than the AMK/AFMK pathway, and comprises >5% of the melatonin-related material detected in mouse urine. 3-HMEL was largely found as a sulfate conjugate. These studies establish sensitive assays for determination of six melatonin metabolites in mouse urine and confirm the potential for antioxidant activity of melatonin through the identification in vivo of AMK and AFMK, ring-opened metabolites with a high capacity for scavenging reactive oxygen species.

Acromioclavicular joint cyst: nine cases of a pseudotumor of the shoulder

OBJECTIVE: (1) To analyse the imaging appearances of nine patients with acromioclavicular joint cysts presenting as shoulder masses for tumor staging with operative, histopathological and joint aspiration findings.DESIGN AND PATIENTS: Retrospective review of imaging and correlation with clinical, operative and surgical notes. Images were reviewed by two musculoskeletal radiologists by consensus. Nine patients who presented clinically with a shoulder mass were evaluated by radiographs (n=9), ultrasound (n=1), conventional arthrography (n=3), MRI (n=6; with direct MR arthrography n=2, indirect MR arthrography n=4).RESULTS: All patients had a focal mass superior to the AC joint, with a size ranging from 1.5 cm to 6 cm and a mean of 3.27 cm. Correlation was available with surgery (n=7), histopathology (n=2) and cyst aspiration (n=2). Two patients were managed conservatively. Geyser sign was positive in all three arthrograms. All MRIs revealed extensive rotator cuff tears with a column of fluid extending from the glenohumeral joint through the rotator cuff tear into the acromioclavicular joint and acromioclavicular cyst. Chondrocalcinosis was seen in the acromioclavicular joint cyst (n=2) and in the glenohumeral joint (n=1). Aspirate in two patients contained calcium pyrophosphate dihydrate crystals.CONCLUSION: Acromioclavicular joint cysts may present as a tumor mass. They are associated with extensive rotator cuff tears and there is usually communication of the cyst with the joint space. This feature excludes a diagnosis of tumor. AC joint cysts may be associated with calcium pyrophosphate dihydrate deposition disease.

Rapid determination of gemcitabine in plasma and serum using reversed-phase HPLC

Gemcitabine (2'2'-difluorodeoxycytidine) is a pyrimidine analog used in the treatment of a variety of solid tumors. After intravenous (i.v.) administration, it is rapidly inactivated to 2'-deoxy-2',2'-difluorouridine (dFdU). A sensitive analytical method for the quantitation of gemcitabine is required for the assessment of alternative dosage and treatment schemes. A rapid and robust RP-HPLC assay for analysis of gemcitabine in human and animal plasma and serum was developed and validated using 2'-deoxyuridine (dU) and 5-fluoro-2'-deoxyuridine (5FdU) as internal standards. It is based on protein precipitation, the use of an Atlantis dC18 column of 100 mm length (inner diameter, 4.6 mm; particle size, 3 microm) and isocratic elution using a 10 mM phosphate buffer, pH 3.0, followed by isocratic elution with the same buffer containing 3% of ACN. For gemcitabine, RSD values for intraday and interday precision were < 4.4 and 5.3%, respectively, the LOQ was 20 ng/mL, and the assay was linear in the range of 0.020-20 microg/mL with an accuracy of > or =89%. The recovery for gemcitabine, dU and 5FdU was 86-98%. The assay was applied to determine gemcitabine levels in plasma samples of patients collected during and shortly after conventional infusion of 25-30 mg/kg body mass (levels: 2.0-18.9 microg/mL) and rats that received lower doses (1.5 mg/kg) via i.v., subcutaneous and oral drug administration (levels: 0.20-2.60 microg/mL). It could also be applied to estimate dFdU levels in human plasma.

Tracing the source of groundwater for three different coastal peatlands along Lake Superior

The goal of this project was to investigate the influence of a large inland lake on adjacent coastal freshwater peatlands. The specific aim was to determine the source of groundwater for three differently formed peatlands located on the southern shore of Lake Superior. The groundwater study was conducted at Bete Grise, a peatland complex in a dune-swale system; Pequaming, a peatland developed in the swale of a tombolo; and Lightfoot Bay, a peatland developed in a barrier beach wetland complex. To determine the source of groundwater in the peatlands, transects of six groundwater monitoring wells were established at each study site, covering distinctly different vegetation zones. At Pequaming and Lightfoot Bay the transects monitored two vegetation zones: transition zone from upland and open fen. At Bete Grise, the transects monitored dunes and swales. Additionally, at all three sites, upland groundwater was monitored using three wells that were installed into the adjacent upland forest. Biweekly measurements of well water pH and specific conductance were carried out from May to October of 2010. At each site, vegetation cover, peat depths and surface elevations were determined and compared to Lake Superior water levels. From June 14 – 17, July 20 – 21 and September 10 – 12, stable isotopes of oxygen (18O/16O) ratios were measured in all the wells and for Lake Superior water. A mixing model was used to estimate the percentage of lake water influencing each site based on the oxygen isotope ratios. During the sampling period, groundwater at all three sites was supported primarily by upland groundwater. Pequaming was approximately 80 % upland groundwater supported and up to 20 % Lake water supported in the uppermost 1 m layer of peat column of the transition zone and open fen. Bete Grise and Lightfoot Bay were 100 % upland groundwater supported throughout the season. The height of Lake Superior was near typical levels in 2010. In years when the lake level is higher, Lake water could intrude into the adjacent peatlands. However, under typical hydrologic conditions, these coastal peatlands are primarily supported by upland groundwater.