Made available in DSpace on 2017-07-10T15:14:33Z (GMT). No. of bitstreams: 1 DissertacaodelvanBlanco.pdf: 1724467 bytes, checksum: f790bc7ea9e5fad4db1ebecded618e88 (MD5) Previous issue date: 2015-03-12 / The increase in productivity of soy beans in recent years happened, basically, for the adoption of technologies such as the use of inputs and equipments, which are directly related to the cost of production. Among the inputs used, fertilizers are more expensive. The western Parana has a large number of pigs and poultry integrators, generating a lot of animal waste. The use of animal waste as fertilizer is a rational alternative of great interest in terms environmental, economic, social and agronomic. Thus the objective of this study was evaluate the effects of different doses application of poultry litter and pig slurry, in to the macronutrient content into soil and soy bean leaves as on grains and soy bean oil compared to mineral fertilization. The experimental area is located at the Experimental Station of the Agronomic Institute of Paraná, in Santa Tereza do Oeste - PR. The treatments were generated by factorial (2x3)+1+1, being two waste (liquid swine and poultry litter) with three doses of each (poultry litter: 1.2, 2.4 and 3.6 t / ha (wet basis) and pig slurry: 48, 96 and 144 m3 (cubic meter) / ha). Others additional treatments were the control (no fertilization) and mineral fertilization (300 kg/ha of the formulated 04-30-10). Experimental delineation adopted was random blocks with three repetitions. The animal manure fertilization resulted in higher levels of P and K into soil and greater absorption of N and P by plants, beyond a higher percentage of lodged plants in relation to the control and mineral fertilization. Grains productivity and soy bean oil income were similar between animal waste and mineral fertilization and higher than the control. It demonstrated both fertilizations are effective providing nutrients for the soy bean culture. / O aumento da produtividade de soja nos últimos anos ocorreu, basicamente, pela adoção de tecnologias como o uso de insumos e equipamentos, o que está diretamente relacionado ao custo de produção. Dentre os insumos utilizados, os fertilizantes são os mais onerosos. A região Oeste do Paraná possui grande número de integradores de suínos e aves, gerando uma grande quantidade de dejetos. A utilização de dejetos de animais como fertilizantes é uma alternativa racional, de grande interesse em termos ambientais, econômicos, sociais e agronômicos. Desse modo, o objetivo deste trabalho foi avaliar os efeitos da aplicação de diferentes doses de cama de aviário e dejetos de suínos, nos teores de macronutrientes no solo e nas folhas de soja, bem como sobre a produtividade de grãos e óleo, em comparação à adubação mineral. A área experimental localiza-se na Estação Experimental do Instituto Agronômico do Paraná, no município de Santa Tereza do Oeste - PR. Os tratamentos foram gerados pelo esquema fatorial (2x3)+1+1, sendo dois dejetos (líquido de suínos e cama de aviário) e três doses de cada (cama de aviário: 1,2; 2,4 e 3,6 t/ha (base úmida) e dejeto líquido de suínos: 48; 96 e 144 m3/ha). Os tratamentos adicionais utilizados foram a testemunha (sem adubação) e o com adubação mineral (300 kg/ha do formulado 04-30-10). O delineamento experimental adotado foi o de blocos ao acaso, com três repetições. A adubação com dejetos de animais resultou em maior teor de P e K no solo e maior absorção de N e P pelas plantas, além de um maior porcentual de plantas acamadas em relação à testemunha e à adubação mineral. A produtividade de grãos e o rendimento de óleo foram equivalentes, entre adubação com resíduos animais e adubação mineral e superiores à testemunha, demonstrando que ambos são eficazes em fornecer nutrientes para a cultura da soja.

Made available in DSpace on 2017-07-10T15:14:43Z (GMT). No. of bitstreams: 1 Dangela Maria Fernandes.pdf: 4431136 bytes, checksum: c95d44de12c14904f785c2581dd1a253 (MD5) Previous issue date: 2012-02-28 / Coordenação de Aperfeiçoamento de Pessoal de Nível Superior / The intensive pig farming's development promoted the production of residual biomass large amounts, when improperly handled becomes a major environment s pollution sources. Therefore, waste management should be seen as an integral part of system's production pigs and should be included in planning this activity. In this context, aiming to evaluate the residual biomass' management generated in the production system for finishing pigs, Unit Colombari farm, there was a study about the water consumption, feed consumption and residues production during the period October 2010 to February 2011 with the goal of generating productivity indices related to the productive system The gotten results indicated that the rates for the daily water consumption per pig ranged from 1.2 to 2.4 liters, the daily feed intake per animal was 1.90 kg and the manure's daily production per animal appeared in 7.3 liters. It is considered that the productive unit is fitted in an efficient handling characteristic, having efficacy in reducing water consumption in the parameterization of the animal feed and residual biomass' control. / O desenvolvimento da suinocultura intensiva promoveu a produção de grandes quantidades de biomassa residual que, quando manejados inadequadamente, tornam-se uma das principais fontes de poluição do meio. Por isso, o manejo de resíduos deve ser visto como parte integrante do sistema produtivo de suínos, devendo estar incluído no planejamento desta atividade. Nesse contexto, visando avaliar o manejo da biomassa residual gerada no sistema produtivo de suínos em terminação, da Unidade Granja Colombari, realizou-se um estudo quanto ao consumo de água, o consumo de ração e a produção de dejetos, durante o período de Outubro de 2010 a Fevereiro de 2011, com o objetivo de gerar índices de produtividade relacionados ao sistema produtivo. Os resultados obtidos indicaram que os índices relativos ao consumo de água diário por suíno variou de 1,2 a 2,4 litros, o consumo de ração diário por suíno foi de 1,90 kg e a produção diária de dejetos por suíno apresentou-se em 7,3 litros. Considera-se que a unidade produtiva se enquadrou nas características de um manejo eficiente, possuindo eficácia na minimização do consumo de água, na parametrização da nutrição animal e no controle de produção de biomassa residual.

Poor egg quality is the leading cause of pregnancy loss and Down's syndrome. While even eggs in young women frequently contain an incorrect number of chromosomes and are therefore unlikely to give rise to a viable pregnancy, the incidence of chromosomally abnormal eggs increases strikingly with advancing maternal age. Why egg quality declines dramatically as women approach their forties remains one of the outstanding questions in developmental biology. This PhD thesis demonstrates how unforeseen features of kinetochore organization that are unique to meiosis render this cell division process in mammals particularly prone to errors. Firstly, my results uncovered an unexpected multi-subunit organization of the meiotic kinetochore, which is widely conserved across mammals and biases eggs towards errors. Secondly, I identified two independent mechanisms that predispose eggs from older women to aneuploidy. The first mechanism affects the fidelity of meiosis I. My analysis revealed that human oocytes challenge the paradigm that sister kinetochores are fully fused. Instead, I demonstrated that sister kinetochores disjoin as women get older, which promoted erroneous kinetochore-microtubule attachments. This in turn allowed chromosomes to rotate on the spindle and provided a mechanistic explanation for reverse segregation - a recently discovered meiotic error that is unique to humans. Secondly, I pioneered the use of super-resolution microscopy to study chromosome architecture in human eggs and discovered that individual kinetochores during meiosis II in mammals are composed of previously unidentified subdomains. In young females, these subdomains are joined together by cohesin complexes. With age, kinetochores fragment into two pieces. Fragmented kinetochores frequently attach merotelically to spindle microtubules, which predisposes aged eggs to errors. What severely hinders our progress in identifying causes of human infertility is that numerous features of human meiosis are not represented in mice. To overcome this challenge, I developed an experimental platform to mimic the age-related changes that occur in humans in oocytes from young mice. I achieved this by extending the applications of Trim-Away, a novel method to degrade endogenous proteins even in primary cells, to partially deplete proteins. Furthermore, I established a new experimental model system to study human-like aspects of meiosis in live non-rodent cells in real time: pig oocytes. Together, these results set foundations for new therapeutic approaches to extend reproductive lifespan by counteracting the age-related loss in kinetochore integrity that this study identified. Furthermore, partial Trim-Away and studying meiosis in pigs opens new directions for meiotic research.

Every fall, Eduardo Zayas-Bazán, a native of Cuba who was a veteran of the 1961 Bay of Pigs invasion and who taught Spanish at East Tennessee State University for over 30 years, hosts a Cuban-style pig roast in Tennessee for family and friends. "Among my fondest memories are the pig roasts we had in Cuba on special occasions," said Zayas-Bazán, who marinates his pigs in grapefruit juice mixed with garlic and oregano, and cooks them in a contraption that he says "looks like a shoe box with a grill inside."" With the annual pig roast as context, the film tells the story of Zayas-Bazán's life, both in Cuba and the U.S. / https://dc.etsu.edu/etsu_books/1098/thumbnail.jpg

Cystic fibrosis (CF) is the most common autosomal recessive genetic disease in Caucasian populations. CF affects multiple organ systems including pancreas, liver, intestines, sweat glands, and male reproductive organs, however the leading cause of morbidity and mortality in CF patients is chronic lung disease. CF is caused by a mutant cystic fibrosis transmembrane conductance regulator (CFTR) gene which leads to chloride (Cl-) and bicarbonate (HCO3-) anion dysregulation at the airway surface. Without adequate anion exchange, thick, viscous mucus accumulates at the airway surface allowing bacterial colonization to occur. Complementing CFTR in the appropriate airway cells restores the anion channel activity in CFTR-deficient cells. The ultimate goal for CF gene therapy is to design an integrating vector that would lead to persistent and efficient expression of CFTR in the airways. Performing gene therapy experiments is dependent upon a relevant animal model. The CF pig is a large animal model similar in size, anatomy, and physiology to humans. Importantly, the CF pig recapitulates human lung disease. From the CF pig, we have learned much about CF lung disease and have developed relevant assays to measure anion channel correction. We have learned that loss of CFTR leads to a decreased airway surface ASL pH, bacterial killing ability, and increased mucus viscosity. Standardized assays have been developed to evaluate the change in current by Ussing chambers, ASL pH, bacterial killing in vivo and ASL pH and viscosity on primary airway cultures in vitro. Ultimately, these metrics allow us to make conclusions about the efficiency of CFTR restoration. Viral vectors are promising candidates for CF gene therapy. Viral vectors such as adenovirus (Ad), adeno-associated virus (AAV), and pseudotyped lentiviral vectors such as feline immunodeficiency virus (FIV) or human immunodeficiency virus (HIV) can efficiently transduce airway cells and express CFTR. Ad and AAV have both been tested in CF clinical trials, but CFTR expression was transient, if detected at all. Understanding vector biology and overcoming barriers in the lung have allowed us to improve vector delivery to the airways. However, the next major hurdle was achieving persistent expression. Ad and AAV are both transiently expressing vectors, and vector readministration is implausible due to the presence of neutralizing antibodies that develop against the vector. Creating a hybrid nonviral/viral vector in which the integrating nonviral piggyBac transposon system is delivered by an Ad or AAV vector has allowed us to achieve persistent expression in mice. In a third integrating vector system, lentiviral vectors have historically been challenging to work with due to low titer levels. However, improvement in vector purification methods have allowed us to validate a lentiviral vector as a viable gene therapy option. In total, we have validated three integrating vector systems by restoring CFTR to CF pigs to correct the phenotypic defect.

Cancer is the second deadliest disease in the United States with an estimated 1.69 million new cases in 2017. Medical imaging modalities, such as computed tomography (CT) and magnetic resonance imaging (MRI), are widely used in clinical medicine to detect, diagnose, plan treatment, and monitor tumors within the body. Advances in imaging research related to cancer assessment have largely relied on consented human patients, often including varied populations and treatments. Tumor bearing mouse models have been highly valued for basic science research, but imaging focused applications are limited by the translational ability of micro imaging systems. Pig models are well suited to bridge the gap between human cohorts and mouse models due to similar anatomy, physiology, life-span, and size between pigs and humans. These models provide the opportunity to advance medical imaging while simultaneously characterizing progressive changes resulting from an intervention, exposure, or genetic modification. We present a foundation for effectively characterizing disease models in pigs, susceptible to tumor development, using longitudinal medical image acquisition and post-processing techniques for quantification of disease. Longitudinal, whole-body protocols were developed with CT and MRI. Focus was placed on systematic process, including transportation, anesthesia and positioning, imaging, and environmental controls. Demonstration of the methodology was achieved with six pigs (30-85 kg) with four to seven imaging time points acquired per animal. Consistent positioning across time points (CT to CT) and within time points (CT to MRI) was assessed with distance measures obtained from the skeleton following rigid registration between images. Alignment across time points was achieved with an average value of 16.51 (± 12.46) mm observed all acquired measurements. For consistent, retrievable, and complete qualitative assessment of acquired images, structured reports were developed, including assessment of imaging quality and emphasis on tumor development throughout the body. Reports were used to perform a systematic, semi-qualitative comparison of CT and MRI lung assessment with an overall agreement of 72% in detection of disease indicators. A multi-level registration algorithm was developed to align anatomic structures of interest in the acquired longitudinal datasets. The algorithm consisted of initialization followed by repeated application of a core registration framework as the input data reduced in image field of view. It was applied to align regions of interest in the brain, upper right lung, and right kidney. Validation was performed with overlap (range = [0.0,1.0], complete overlap = 1) and distance measures (range = [0.0, ∞], perfect match = 0.0) of corresponding segmentations with overall results of 0.85 (± 0.11) and 0.41 (± 0.83) mm, respectively. An extension of the algorithm was created, demonstrating the ability to incorporate directional growth and feature extraction measurements into longitudinal tumor progression monitoring. Techniques were applied to a phantom dataset showing solid tumor growth and transition from a non-solid to part-solid lesion in the lungs. Finally, the developed methods – imaging, structured reporting, registration, and longitudinal feature extraction – were applied to four different porcine models pre-disposed to tumor development. 1) A genetically modified Li-Fraumeni (TP53R167H/+/TP53R167H/R167H) background model showing the development of osteosarcoma and lymphoma. 2) A TP53R167H/+ animal with exposure to crystalline silica showing progression of silicosis in the lungs. 3) TP53R167H/+/TP53R167H/R167H animals with exposure to radiation for targeted sarcoma development and 4) TP53R167H/+ pigs with conditional KRASG12D/+ mutation activated in the lung and pancreas. Whole-body and targeted imaging protocols were developed for each model and qualitatively interpreted by a radiologist using structured reports. Multi-level registration was used to align identified tumors and longitudinal features were extracted to quantitatively track change over time. Overall, the developed methods aided in the effective, non-invasive characterization of these animals.

The initial focus of this PhD project was on comparative gene mapping. Comparative gene mapping is facilitated by consensus PCR primers which amplify homologous gene fragments in many species. As a part of an international co-ordinated programme of comparative mapping in pigs, 47 CATS (Comparative Anchor Tagged Sequence) consensus primer pairs for loci located on human chromosomes 9, 10, 20, and 22, were used for amplifying homologous loci in pigs. After optimization of PCR conditions, 23 CATS products have confirmed by comparison with homologous sequences in GenBank. A French somatic cell hybrid panel was used to physically map the 6 porcine CATS products distinguishable from rodent background product, namely ADRA1A, ADRA2A, ARSA, GNAS1, OXT and TOP1. Of these, the map location of ADRA1A and OXT showed inconsistency with the previously recognised conserved relationship between human and pig. The other four loci mapped to positions consistent with known syntenic relationships. Despite low levels of polymorphism, frequently indistinguishable rodent and porcine products in somatic hybrids and some confusion of identity of gene family members, these CATS primers have made a useful contribution to the porcine-human comparative map. The focus of the project then changed to genetic and molecular characterisation of endogenous retroviruses in pigs and their relatives. Pigs are regarded as a potentially good source of organs and tissues for transplantation into humans. However, porcine endogenous retroviruses have emerged as a possible problem as they can infect cultured human cells. Two main types of pig retrovirus, determined by envelope protein, PERV-A and PERV-B, are widely distributed in different pig breeds and a third less common type, PERV-C, has also been recognised. Endogenous retroviruses were analyzed from the Westran (Westmead transplantation) inbred line of pig, specially bred for biomedical research. Thirty-one 1.8 kb env PCR product clones were sequenced after preliminary screening with the restriction enzymes KpnI and MboI. Five recombinant clones between A and B were identified. 55% of clones (17/31) sequenced had stop codons within the envelope protein-encoding region, which would prevent the retrovirus from making full-length envelope protein recognizable by cell-surface receptors of the virus. The endogenous viruses were physically mapped in Westran pigs by FISH (Fluorescence In Situ Hybridisation) using PERV-A and PERV-B envelope clones as probes. Preliminary FISH data suggest that there are at least 22 PERVs (13 PERV-A and 9 PERV-B) and the chromosomal locations of these in the Westran strain are quite different from European Large White pigs. The sequences and mapping results of inbred Westran pig suggest that there are relatively few PERV integration sites compared with commercial pigs and further that a large proportion of clones are defective due to premature stop codons in the envelope gene. To investigate the relationship of endogenous retroviruses in peccaries and pigs, a set of degenerate primers was used to amplify peccary retroviral sequences. The sequences of two putative retroviral clones showed close homology, albeit with a 534 bp deletion, to mouse and pig retroviral sequences. Also, four non-target sequences were amplified from peccary with the degenerate retroviral primers. They are a part of the peccary cofilin gene, a SINE, and a sequence containing a microsatellite. The peccary endogenous retroviral sequences are significant in that they are the first such sequences reported in peccary species and repudiate old claims in the literature that peccaries do not have C-type retroviral sequences.

Immunological stressors, in the form of clinical and sub-clinical disease are currently controlled using both prophylactic antibiotics in-feed, and therapeutic antibiotic treatment. Respiratory disease, primarily Actinobacillus pleuropneumoniae (App) infection, is recognised as a major factor causing reduced productivity in pigs. This thesis reports investigations into the use of novel immunomodulators in particular Interleukin 4 (IL-4) and Interleukin 1 receptor antagonist (IL-1ra) as alternatives to antibiotics to treat App infection. Immunological and molecular biological assays were used to investigate and accumulate data. An in vitro study undertaken to find potential anti-inflammatory substances, revealed that Interleukin 8 (IL-8) mRNA production stimulated by PMA or LPS in whole pigs' blood was suppressed by IL-4. IL-1ra also suppressed stimulated IL-8 mRNA production by heat killed App bacteria (KB) in vitro. An acute LPS challenge in pigs in vivo however, showed no variation in illness or weight loss between pigs treated prophylactically with anti-inflammatory substance (IL-4 and IL-1ra) and saline treated pigs. The use of plasmids as a delivery system for anti-inflammatory substance did not show promise since it did not enhance growth or prolong the expression of the substances in the pigs. However, in the chronic App challenge model IL-4 and IL-1ra administered prophylactically in vivo showed an ability to improve growth. The therapeutic administration of IL-4 and IL-1ra to App challenged pigs showed no difference in pigs' growth, regardless of the treatment or control administered. To conclude, IL-4 and IL-1ra showed promise when administered prophylactically and improved growth and abrogated disease under conditions of App challenge. However when IL-4 and IL-1ra where administered therapeutically they did not perform as well. Moreover these compounds have potential as a commercial application to reduce the growth reduction caused by disease such as App.

<p>Septic shock is frequently seen in intensive care units and is associated with significant mortality. Endotoxin – a major mediator of the pathophysiologic responses – is released during lysis of Gram-negative bacteria. These responses can be mimicked in the endotoxaemic pig.</p><p>This thesis focuses on the following topics: the inflammatory and pathophysiological responses to various endotoxin doses and infusion patterns; covariations between endotoxin induced inflammatory and pathophysiological responses; whether the biological effects of endotoxin can be modulated by clopidogrel and whether tobramycin or ceftazidime reduce plasma cytokine levels.</p><p>Endotoxin induced linear log-log cytokine and F2-isoprostane responses. Leukocyte and platelet responses, pulmonary compliance, circulatory variables as well as indicators of plasma leakage and hypoperfusion exhibited log-linear responses to the endotoxin dose. Biological responses to endotoxaemia such as inflammation, hypotension, hypoperfusion and organ dysfunction were more expressed when the organism was exposed to endotoxin at a higher rate. These results may facilitate the possibility to choose relevant endotoxin administration, when experiments are set up in order to evaluate certain responses to endotoxaemia.</p><p>Correlation studies between cytokines, leukocytes, platelets and the endotoxin dose were in agreement with the well-known ability of endotoxin to induce cytokine expression and to activate both primary haemostasis and leukocytes. Free radical mediated lipid peroxidation and COX-mediated inflammation correlated to cytokine expression and organ dysfunction in endotoxaemic shock. </p><p>Endotoxaemic pigs pretreated with clopidogrel, exhibited a trend towards less expressed deterioration of renal function, although blocking of ADP-induced primary haemostasis is not a key mediator of endotoxin induced deterioration of renal function.</p><p>Tobramycin did not neutralise the biological effects of endotoxin or the plasma levels of endotoxin, suggesting that these antibiotics do not bind to endotoxin.</p><p>Reduction in IL-6 was greater in pigs treated with ceftazidime and tobramycin as compared with those given saline, indicating a possible anti-inflammatory effect of both antibiotics.</p>

Pig follicle development is a complex but poorly understood process involving both gonadotrophins and local ovarian factors. A series of studies sought to investigate these intrafollicular cell-to-cell interactions. Microarray analysis combined with gene ontology revealed that the oocyte, granulosa and theca cells each expressed more than 650 potential secreted factors and receptors, including members of the TGF-β, IGF1, EGF and FGF families. Using a well-defined in vivo experimental paradigm that generates follicles and oocytes of different quality, the temporal expression of several growth factors in the oocyte, granulosa and theca cells collected from sows during the recruitment and mid-selection phases, as well as the final selection of the preovulatory follicle population before and after the LH surge was studied. IGF1 expression patterns indicated its potential for modulating granulosa and theca cell function during the selection stages, and an involvement in creating differences in follicular quality between the first and second preovulatory wave post-weaning. Transient up-regulation of AREG and EREG mRNA around the LH surge, suggested their involvement in ovulation. Results of a second study investigating TGF-β superfamily expression, suggested a role for GDF9 in follicle selection through the up-regulation of TGFBR1 expression, while BMP15 could be involved in ovulation through the up-regulation of BMPR1B. Expression of angiogenesis-related genes during follicle development was also investigated. During mid-selection, ANGPT2 may allow VEGFA and similar factors to stimulate vascular development or destabilize the vasculature and cause follicular atresia, while ANGPT1 may be required in the preovulatory follicle population. Associations among the transcription factor HIF1A, VEGFA and ANGPT1, suggested interactions between the ligands in regulation of angiogenesis. Finally, the effects of the pig oocyte on cumulus cell function was assessed by co-culturing cumulus cell complexes with or without denuded oocytes isolated from large oestrogenic follicles. Presence of oocytes decreased FSHR and increased HSD3B expression, potentially stimulating progesterone while attenuating oestradiol production. In conclusion, oocytes were shown to control cumulus cell function in a way that reflects their specific environment and further evidence was obtained for a complex network of growth factors and receptors in the follicle and their involvement in regulation of pig follicle development. / Animal Science