L'epatite E è una malattia umana con caratteristiche di epatite acuta, causata da un ssRNA virus (HEV). Nel 1997, HEV è stato identificato per la prima volta nei suini (SwHEV). In seguito, diverse evidenze, tra cui la vicinanza genetica tra ceppi umani e suini, suggerirono la trasmissione zoonotica del virus. Nella presente tesi, l’identificazione di SwHEV è stata condotta mediante ricerca di porzioni di genoma virale attraverso RT-PCR. Dal 2011 al 2013, sono stati analizzati 343 campioni fecali (da 19 allevamenti) e 70 bili (da 2 macelli) prelevati da altrettanti suini, in diverse Regioni italiane. E’ stato inoltre condotto uno studio retrospettivo su 78 feci (da 3 allevamenti) raccolte nel 2000. Il virus è stato identificato nel 24,5% e 19,2% delle feci raccolte rispettivamente nel 2011-2013 e nel 2000. Nessuna bile è risultata positiva. Mediante sequenziamento del genoma intero di uno dei virus identificati, è stata condotta l’analisi filogenetica per valutarne il grado di correlazione con alti ceppi suini e umani. La presenza di HEV è stata valutata lungo la filiera di produzione suina, dal macello al punto vendita. Trentaquattro campioni di feci, fegato e muscolo sono stati raccolti in un macello da altrettanti suini sani (età:6-7 mesi). Quattordici feci e 2 fegati, sono risultati positivi per HEV. Sono state prelevate 129 salsicce sia allo stabilimento di trasformazione sia alla vendita, ma nessuna è risultata positiva. La presenza di HEV è stata valutata anche nelle salsicce di fegato, fresche e secche, acquistate presso una macelleria. Il genoma virale è stato rilevato nel 22,2% delle salsicce fresche e nel 4,3 % di quelle secche ma la vitalità del virus non è stata dimostrata. In conclusione, lo studio condotto ha confermato l’ampia circolazione di HEV nei suini e la possibile contaminazione dei prodotti carnei derivati, confermando la necessità di una continua sorveglianza. / Hepatitis E is an acute disease of humans, caused by a positive ss RNA virus, Hepatitis E virus (HEV). Several evidences, including genetic similarity between human and swine HEV strains from the same area, suggest zoonotic transmission. From 2011 to 2013, we conducted a survey for HEV in 19 swine farms and 2 slaughterhouses in different Italian regions, collecting 343 feces and 70 bile samples. Moreover, a retrospective study was conducted on 78 animals housed in 3 farms locate in Northern Italy and collected in 2000. Overall HEV was detected in 24.5% and 19.2% of fecal samples collected in 2011-2013 and in 2000, respectively. On the contrary, none of bile samples was positive. Furthermore, the genome of one SwHEV strain was completely sequenced to evaluate possible correlation with swine and human HEV strains. We investigated presence of HEV in a swine food chain from farm to finish products, enrolling a big Italian factory which trades pig meat across the country. Thirty-four feces, liver and muscle samples were collected in a slaughterhouse from 34 clinically healthy pigs, aged 6-7 month old. At slaughterhouse 14 fecal samples and 2 liver, resulted positive for HEV. Moreover, none of the sausages (129) collected at processing and point of sale were positive. We have also evaluated the presence of HEV and fecal cross-contamination in pork liver sausages bought at a grocery in Italy. The genome of HEV was detected in both raw (22.2%) and dry (4.3%) liver sausages, but the viability of the virus was not demonstrated. Finally, several cell lines were inoculated with HEV-positive fecal and pork liver samples to attempt HEV replication but virus growth was not established.

In veterinary medicine, the ability to classify mammary tumours based on the molecular profile and also determine whether the immunophenotype of the regional lymph node and/or systemic metastases is equal to that of the primary tumor may be predictive on the estimation of the effectiveness of various cancer treatments that can be scheduled. Therefore, aims, developed as projects, of the past three years have been (1) to define the molecular phenotype of feline mammary carcinomas and their lymph node metastases according to a previous modified algorithm and to demonstrate the concordance or discordance of the molecular profile between the primary tumour and lymph node metastasis, (2) to analyze, in female dogs, the relationship between the primary mammary tumor and its lymph node metastasis based on immunohistochemical molecular characterization in order to develop the most specific prognostic-predictive models and targeted therapeutic options, and (3) to evaluate the molecular trend of cancer from its primary location to systemic metastases in three cats and two dogs with mammary tumors. The studies on mammary tumours, particularly in dogs, have drawn gradually increasing attention not exclusively to the epithelial component, but also to the myoepithelial cells. The lack of complete information on a valid panel of markers for the identification of these cells in the normal and neoplastic mammary gland and lack of investigation of immunohistochemical changes from an epithelial to a mesenchymal phenotype, was the aim of a parallel research. While investigating mammary tumours, it was noticed that only few studies had focused on the expression of CD117. Therefore, it was decided to further deepen the knowledge in order to characterize the immunohistochemical staining of CD117 in normal and neoplastic mammary tissue of the dog, and to correlate CD117 immunohistochemical results with mammary histotype, histological stage (invasiveness), Ki67 index and patient survival time.

Toxoplasma gondii is an obligate intracellular parasite capable of infecting virtually all warm-blooded species, including humans, but cats are the only definitive hosts. Humans or animals acquire T. gondii infection by ingesting food or water contaminated with sporulated oocysts or by ingesting tissue cysts containing bradyzoites. Toxoplasmosis has the highest human incidence among zoonotic parasitic diseases, but it is still considered an underreported zoonosis. The importance of T. gondii primary infection in livestock is related to the ability of the parasite to produce tissue cysts in infected animals, which may represent important sources of infection for humans. Consumption of undercooked mutton and pork are considered important sources of human Toxoplasma gondii. The first aim of this thesis was to develop a rapid and sensitive in- house indirect ELISA for the detection of antibodies against T. gondii in sheep sera. ROC-curve analysis showed high discriminatory power (AUC=0.999) and high sensitivity (99.4%) and specificity (99.8%) of the method. The ELISA was used to test a batch of sheep sera (375) collected in the Forli-Cesena district. The overall prevalence was estimated at 41.9% demonstrating that T. gondii infection is widely distributed in sheep reared in Forli-Cesena district. Since the epidemiological impact of waterborne transmission route of T.gondii to humans is now thought to be more significant than previously believed, the second aim of the thesis was to evaluate PCR based methods for detecting T. gondii DNA in raw and finished drinking water samples collected in Scotland. Samples were tested using a quantitative PCR on 529 bp repetitive elements. Only one raw water sample (0.3%), out of the 358 examined, tested T. gondii positive demonstrating that there is no evidence that tap water is a source of Toxoplasma infection in Scotland.

La Sindrome da Asfissia Perinatale (PAS) è una delle più comuni patologie che colpiscono il puledro neonato nelle prime 72 h di vita. È una patologia difficile da diagnosticare in quanto non esistono parametri o segni clinici specifici, la sintomatologia è molto variabile in base alla durata e all’intensità dell’insulto ipossico ischemico e al tipo di organo maggiormente colpito. Lo scopo di questo studio è la ricerca e la valutazione di alcuni parametri biochimico-clinici e di alcuni biomarkers per la diagnosi precoce e il corretto trattamento dei puledri affetti da PAS. Nei puledri neonati che presentano questa patologia è stata riscontrata un’ipermagnesiemia al momento del ricovero associata a prognosi infausta, probabilmente causata da un grave danno cellulare con rilascio in circolo del magnesio intracellulare. La PAS potrebbe essere un’ulteriore causa di Euthyroid Sick Syndrome, in quanto abbiamo riscontrato una diminuzione delle concentrazioni di T3 e T4 nei puledri malati rispetto ai sani della stessa età, come avviene in altre malattie sistemiche. Lo studio del profilo proteomico ha permesso di separare le più importanti frazioni proteiche del liquido amniotico di cavalla, mettendo in evidenza similitudini e differenze qualitative e quantitative nei ferogrammi dei puledri sani e di quelli affetti da PAS ed una maggiore variabilità è stata riscontrata nei profili dei liquidi amniotici dei puledri malati. Il glutatione è risultato poco espresso nel puledro neonato, i puledri sani presentano concentrazioni più basse sia rispetto ai malati della stessa età sia agli adulti ma con una tendenza all’aumento nelle prime 24 ore di vita per i sani ed un calo nei malati. La somministrazione della terapia antiradicalica non influisce sulle concentrazioni di glutatione totale ed i puledri deceduti presentano concentrazioni più alte. / Perinatal Asphyxia Syndrome (PAS) that occurs during the perinatal period is one of the most common diseases affecting foals within the first 72 h of life. The diagnosis of PAS is very difficult and relies on an accurate history, identification of neurological signs, and exclusion of other cause of neurological deficits. No specific clinical-pathological and laboratory findings have been highly suggestive of neonatal asphyxia, many of the noted clinical signs can occur with other clinical conditions and depending on the duration and the intensity of hypoxic insult and the tissue injury. The aim of this study was to discover Perinatal Asphyxia Syndrome biomarkers in neonatal foals having an early identification and interventions of the foals at highest risk developing this syndrome. Foals affected by PAS present hypermagnesaemia at admission compared to healthy foals, may be the results of serious tissue damage with cell death and release of intracellular Mg. Higher concentrations is associated to poor outcome. Data obtained in this trial suggest that PAS may cause lower T3 and T4 concentrations in affected foals than in age-matched healthy foals, as reported for other systemic illnesses. PAS could be a cause of Euthyroid Sick Syndrome. The differences revealed between electrophoretic patterns of healthy and sick foals amniotic fluids have showed a qualitative and quantitative variability in the proteomic profile. We found a high proteomic profiling variability also among sick foals. The acute period of hypoxia-ischemia is followed by a period of reperfusion, the second tissue injury occurs in this phase. The cell damage is caused by post-ischemic release of oxygen radicals. Glutathione (endogenous antiradical) concentration (tGSH) is lower in healthy foals compared to sick foals and adults. The antiradical therapy didn’t change the tGSH concentration and no-surviving foals had higher concentration.

This was a retrospective study including ninety samples of dogs with a histological diagnosis of intermediate grade cutaneous mast cell tumour (MCT). The objectives of the study were to validate Minichromosome Maintenance Protein 7 (MCM7) as a prognostic marker in MCTs and to compare the ability of mitotic index (MI), Ki67 and MCM7 to predict outcome. The median survival for the entire population was not reached at 2099 days. The mean survival time was 1708 days. Seventy-two cases were censored after a median follow up of 1136 days and eighteen dogs died for causes related to the MCT after a median of 116 days. For each sample MI, Ki67 and MCM7 were determined. The Receiver Operating Characteristic (ROC) curve was obtained for each prognostic marker to evaluate the performance of the test, expressed as area under the curve, and whether the published threshold value was adequate. Kaplan-Meier and corresponding logrank test for MI, Ki67 and MCM7 as binary variables was highly significant (P<0.0001). Multivariable regression analysis of MI, Ki67 and MCM7 corrected for age and surgical margins indicated that the higher risk of dying of MCT was associated with MCM7 > 0.18 (Hazard Ration [HR] 14.7; P<0.001) followed by MI > 5 (HR 13.9; P<0.001) and Ki67 > 0.018 (HR 8.9; P<0.001). Concluding, the present study confirmed that MCM7 is an excellent prognostic marker in cutaneous MCTs being able to divide Patnaik intermediate grade tumours in two categories with different prognosis. Ki67 was equally good confirming its value as a prognostic marker in intermediate grade MCTs. The mitotic index was extremely specific, but lacked of sensitivity. Interestingly, mitotic index, Ki67 and MCM7 were independent from each other suggesting that their combination would improve their individual prognostic value.

The nature of objectives of vocational education and training (VET) in Botswana identifies it as a national strategy aimed at augmenting economic growth and development. This assigns importance to investments on VET as a form of investment in human capital targeted at the economy and specifically to meet labour market needs. Thus, educational accountability, prompted by economic, technological and structural changes is of great importance to the major stakeholders of VET. This research examines a VET curriculum, specifically the Botswana Technical Education Programme-building construction curriculum, in regard to the degree of alignment between the curricula and the training needs of the building construction curricula. At the core of the research is the need to investigate the fitness for purpose of the curricula designed to provide skilled manpower to the building and construction industry. The research results indicates that: <ul> <li>-- there is a sharp contrast between the broad occupational needs of the industry and the focus of the written curriculum. The written curriculum is skewed towards the provision of training at the unskilled occupation levels while the human resource needs of the industry are within the technical and professional skilled occupational areas;</li> <li>-- vocational educators have developed the written curriculum at the micro level of the building construction industry, while the industry requires programmes to address macro level goals of the industry;</li> <li>-- the BTEP validation model presented responds to concerns of internal quality control processes in curriculum development (performing various vetting functions on the document). It does not provide overall guidance for alignment strategies to be used to ensure alignment of the curriculum and the training needs of the industry;</li> <li>-- the tension between employers from the industry on the one hand, and vocational educators on the other, stems from criticisms that vocational programmes are out¨Cof-touch with the realities of the industry;</li> <li>-- VET educators are not acting as brokers of government policy. Government has a very clear policy on the role and purpose of VET and has provided a direction for the role of VET in this regard.</li> </ul> Further research needs to be concerned with the wider economic implications of having a supply driven VET sector as opposed to, at minimum, striving to create a balance between the supply and demand side of vocational training. The VET sector in Botswana also needs to be concerned with building industry/education partnerships to address issues of workforce development. Future research needs to identify ways in which VET educators can maintain currency of knowledge and thus keep VET curriculum relevant. Copyright 2005, University of Pretoria. All rights reserved. The copyright in this work vests in the University of Pretoria. No part of this work may be reproduced or transmitted in any form or by any means, without the prior written permission of the University of Pretoria. Please cite as follows: Bennett, S 2005, An investigation into curriculum alignment in building construction curricula, MEd dissertation, University of Pretoria, Pretoria, viewed yymmdd < http://upetd.up.ac.za/thesis/available/etd-09052005-100444 / > / Dissertation (MEd (Curriculum and Instructional Design and Development))--University of Pretoria, 2006. / Curriculum Studies / unrestricted

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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.

Seafood is known for its valuable and healthy nutrients; however scientific studies demonstrated the unavoidable presence of contaminants in fish and shellfish. Authorities strongly recommend fish consumption, especially to sensible groups of population (pregnant or nursing women, infants and children), and ask the scientific community for novel and coherent data to establish clear guidelines for consumers. There is also a serious need to extend the monitoring to emerging chemical pollutants, other than methylmercury, dioxins and polychlorinated biphenyls, to better understand the level of seafood contamination. An innovative UHPLC-MS/MS method for the quantification of glycine betaine (GB) in Tapes philippinarum was developed and validated, taking the 2002/657 European Decision as guideline; GB is an endogenous osmolyte abundant in molluscs, and represent a vital methylating agent in humans. Moreover, a previously developed and validated UHPLC-MS/MS method was used to conduct a preliminary monitoring of the presence of the two main perfluorinated contaminants, perfluorooctane sulfonate (PFOS) and perfluorooctanoic acid (PFOA), in six fish species from Italian markets (Dicentrarchus labrax, Merluccius merluccius, Mugil cephalus, Pleuronectes platessa, Scomber scombrus and Sparus aurata). The daily contribution of seafood to PFOS and PFOA dietary intake in Italian population was calculated and data were compared to the corresponding Tolerable Daily Intakes established by EFSA in 2008. Results revealed that the risk of contamination related to fish consumption is unlikely, but suggested the need to conduct further surveys focused on certain species from different sampling site. Subsequently, a mono-specie monitoring was carried out in 140 farmed and wild subjects of Dicentrarchus labrax, from 14 different locations in the Mediterranean area. The obtained results showed that wild subjects are sensibly more contaminated than farmed ones, and that levels of fish contamination are strongly influenced by their geographical origin and the proximity to polluted areas.

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