Social conflict is a part of everyday life, and it can be a potent stressor for both humans and other animals. In the laboratory, when two Syrian hamsters (Mesocricetus auratus) compete for territory, a dominance hierarchy is quickly formed. Becoming subordinate is a significant stressor resulting in increased release of adrenocorticotropic hormone, β-endorphin, and cortisol. Defeated hamsters will also subsequently fail to display territorial aggression in future social encounters and will instead display increased submissive behavior, even in the presence of a smaller, non-aggressive intruder. This change in behavior is consistent and long-lasting and has been termed conditioned defeat (CD). Corticotropin releasing factor (CRF) is an important neuropeptide in the control of the hypothalamo-pituitary-adrenal (HPA) axis response to stress. It is also involved in a number of behaviors such as anxiety, stress responding, food intake, learning, and memory. The widespread distribution of CRF, CRF-like peptides, and CRF receptors, particularly in brain sites related to anxiety, fear, and stress responses, suggests a role for CRF and CRF-like peptides in modulating emotional responses other than via HPA axis activity. It has also been shown that CRF may have a role in the acquisition and expression of CD. Non-specific and CRF type 2-specific CRF antagonists reduce the acquisition and expression of CD in male hamsters while injection of a CRF type 1-specific antagonist does not. Therefore, the goal of this dissertation was to investigate the role of CRF type 1 and 2 receptors in CD in hamsters and to identify neuroanatomical locations where CRF may be acting. It was found that non-specific or CRF type 1 receptor specific agonists enhance the expression, but not acquisition, of CD. Further, these agonists appear to enhance aggressive behavior in animals that were not previously defeated, suggesting a modulatory role for CRF type 1 receptors in agonistic behavior that depends on an animal’s previous social experience. Further, localization of CRF receptors was determined in hamster brain in sites thought important for CD and agonistic behavior, but changes in receptor binding following defeat were not observed. Implications of these results and future directions are discussed.

Les facteurs de risque individuels et environnementaux associés à la contamination par l'agent de la tremblante classique du mouton et à la durée d'incubation des animaux infectés demeurent mal connus. En l'absence de diagnostic antemortem précoce, la longue durée d'incubation de la maladie et les durées de vie limitées des animaux conduisent à une connaissance partielle du statut sanitaire des animaux. De plus, l'analyse doit tenir compte du fait qu'une partie seulement des animaux se contamine. <br />Nous utilisons des modèles d'analyse des données de survie prenant en compte l'existence d'une fraction non à risque. Nous proposons une approche par vraisemblance pénalisée, qui allie les avantages des modèles paramétriques et semi paramétriques existants. Nous nous intéressons ensuite aux modèles paramétriques de survie avec fraction non à risque et effets aléatoires afin de tenir compte du regroupement des animaux dans les élevages. Ces différentes approches sont évaluées à l'aide d'études de simulations.<br />L'application des ces modèles aux données issues du suivi longitudinal d'élevages infectés des Pyrénées Atlantiques (France) confirme le rôle déterminant du génotype au gène PRP sur le risque de contamination et les durées d'incubation. Nos résultats suggèrent de plus que la contamination par l'agent infectieux a principalement lieu en période néonatale. Enfin la forte hétérogénéité des risques de contamination et des durées d'incubation mise en évidence entre troupeaux pourrait être partiellement expliquée par la prise en compte de la structure génétique des élevages et du nombre d'animaux infectés présents.

Cache Valley virus (CVV) induced malformations have been previously reproduced in ovine fetuses; however, no studies have established the CVV infection sequence of the cells targeted by the virus or the development of the antiviral response of the early, infected fetus that results in viral clearance before development of immunocompetency. To address these questions, ovine fetuses at 35 dg were inoculated in utero with CVV and euthanized at 7, 10, 14, 21 and 28 dpi. On postmortem examination arthrogryposis and oligohydramnios were observed in some infected fetuses. Morphologic studies showed necrosis in the central nervous system (CNS) and skeletal muscle of earlier infected fetuses and hydrocephalus, micromyelia and muscular loss in later infected fetuses. Using immunohistochemistry and in situ hybridization, intense CVV viral antigenic signal was detected in the brain, spinal cord, skeletal muscles and fetal membranes of infected fetuses. Viral signal decreased in targeted and infected tissues with the progression of the infection. To determine specific cell types targeted by CVV in the CNS, indirect immunofluorescence was applied to sections of the CNS using a double labeling technique with antibodies against CVV together with antibodies against neurons, astrocytes and microglia. CVV viral antigen was shown within the cytoplasm of neurons in the brain and spinal cord. No viral signal was observed in microglial cells; however, infected animals had marked microgliosis. The antiviral immune response in immature fetuses infected with CVV was evaluated. Gene expression associated with an innate, immune response was quantified by real-time, quantitative PCR. Upregulated genes in infected fetuses included ISG15, Mx1, Mx2, IL-1, IL-6, TNF-?, TLR-7 and TLR-8. The amount of Mx protein, an interferon stimulated GTPase capable of restricting growth of bunyaviruses, was elevated in the allantoic and amniotic fluid in infected fetuses. ISG15 protein expression was significantly increased in target tissues of infected animals. B lymphocytes and immunoglobulin-positive cells were detected in lymphoid tissues and in the meninges of infected animals. This demonstrated that the infected ovine fetus is able to stimulate an innate and adaptive immune response before immunocompetency that presumably contributes to viral clearance in infected animals.

Burn injuries represent a major cause of mortality and morbidity as well as a significant drain on limited resources, particularly in the developing world. Advances in resuscitation, critical care, protective ventilatory strategies, earlier complete debridement complemented by more aggressive treatment of burn wound sepsis have reduced the mortality of thermal injuries. There has also been a move to focus on education and prevention campaigns, which have borne fruit and resulted in some reduction in the incidence of burns in the paediatric population. Burn care, once a Cinderella specialty, has become a well focused multi-disciplinary specialty in its own right. Burn injury is dissimilar to many forms of trauma. In major burns, the initial trauma may be limited to the skin alone or skin and lungs, but all organ systems are rapidly involved as the physiological derangement becomes systemic. The burden of this multi-system insult is substantial. Globally, the World Health Organisation estimated that fire-associated burns alone directly resulted in over 320,000 deaths in 2002, and in the USA, approximately 1million children sustain burns each year. Australasian guidelines suggest that all adult burns with greater than 15% total body surface area (TBSA) and >10% TBSA burn in children will require fluid resuscitation and possibly critical care support at some point in their hospitalisation. Mortality in these patients with severe burns follows a bimodal pattern of early and late deaths. Causes of early death comprise refractory shock, inability to obtain a safe airway or provide adequate oxygenation, co-existent trauma, non-survivable carboxyhaemoglobin poisoning, and decisions that injuries are non-compatible with recovery, leading to therapy withdrawal. With improved resuscitation strategies, 95% of patients survive the early resuscitation phase. Late deaths are secondary to sepsis normally associated with wound infection and multiple organ failure. The morbidity associated with burn injury continues well after the acute hospital admission, frequently for up to several years post burn injury, as is witnessed by prolonged rise in basal metabolic rate and worsening scar tissue1, 2. It has been calculated that in children approximately 60% of the cost of burn care occurs post wound closure; that is, dealing with the disabling and disfiguring contractures associated with hypertrophic scar3, 4. Hence, research and new modalities are being aimed at reducing cost of treatment and improving quality of life for survivors of burn injury. Thus, this PhD aims to reflect the multidisciplinary nature of modern day burn care, with the inclusion of seven published papers and one book chapter covering prevention and education relevant to paediatric burns, treatment and minimisation of wound infection , and scar minimisation, along with the first study into the relevance of fetal wound healing post burn injury.

[Truncated abstract] In obstetric practice, pregnant women at risk of pre-term delivery between 24 and 34 weeks of gestation are administered synthetic glucocorticoids (betamethasone or dexamethasone) to induce fetal organ maturation. During this gestational period, the fetal kidney is undergoing a phase of rapid organogenesis with an increase in renal growth and active nephrogenesis occurring. The studies comprising this thesis examine the effects of prenatal betamethasone exposure on the fetal and adult ovine kidney. The central hypothesis of these studies was that exposure of the fetal kidney to betamethasone in late gestation would change renal structure and induce long-term alterations in the expression of glucocorticoid-sensitive genes and proteins. In the fetal studies, pregnant Merino ewes bearing single fetuses received single or repeated-weekly intra-muscular (i.m.) injections of betamethasone (0.5 mg/kg body weight) or saline commencing on day 104 of gestation (term is 150 days). Kidneys were collected from fetuses at 109, 116, 121 and 146 days of gestation (d). Using gold standard unbiased stereological techniques, the physical disector/fractionator method, total glomerular (nephron) number and glomerular volume were determined in 146 d fetal kidneys exposed to repeated maternal saline or betamethasone administration. In the adult study, kidneys were collected from 3.5-year-old sheep that had been exposed to ... In this thesis I have demonstrated that renal growth restriction as a result of betamethasone exposure is associated with a reduction in fetal nephron endowment. Although betamethasone does not appear to consistently alter nephron number or glomerular size, it may indirectly affect total nephron endowment through effects on renal growth. I have also provided evidence which suggests that lategestation betamethasone exposure in sheep does not program permanent alterations in the renal expression of genes or proteins involved in glucocorticoid hormone action or components of the renin-angiotensin system. Therefore, exposure of the fetal kidney to betamethasone during nephrogenesis may alter renal structure if kidney growth is perturbed; however, there are no persistent alterations in the expression of glucocorticoid-sensitive genes. These findings are consistent with the preservation of normal basal blood pressure in the adult sheep I studied and with the limited results from human studies of late-gestation maternal glucocorticoid administration.

Orientador: Wilter Ricardo Russiano Vicente / Banca: Luiz Fernando de Souza Rodrigues / Banca: Paulo Henrique Franceschini / Resumo: Foram realizados 20 programas de superovulação e colheita de embriões em ovelhas Santa Inês, com objetivo de testar a hipótese que a adição de LH ao final do protocolo de superovulação causa incremento nas taxas de ovulação e de produção de embriões. Para tanto, o experimento foi delineado em cross-over, sendo o estro sincronizado com CIDR™ por 14 dias. No D7, substituiu-se o implante por um novo, e administrou-se 37,5 μg de d-cloprostenol, IM. Para superestimulação, 256 mg de FSHp foram administradas em 8 doses decrescentes, iniciando no D12. No D14, as fêmeas receberam 200 UI de eCG e 37,5 μg de d-cloprostenol. No grupo controle (G-C) a taxa FSH/LH foi constante, enquanto no G-LH o protocolo preconizava o acrescido de 7,5 mg de LH, administrado 24 h após a retirada do implante (D15). As fêmeas foram inseminadas com sêmen congelado, por laparoscopia, as 42 e 48 h após a retirada do implante, concomitantemente, as estruturas ovarianas foram avaliadas. No D21, os embriões foram cirurgicamente colhidos e avaliados. Os resultados demonstraram que o LH administrado ao final do tratamento superovulatório não incrementou de maneira significativa a taxa de ovulação e o número de embriões viáveis (78,7% ± 22,72 e 3,77 ± 4,29, respectivamente, G-C; 84,75 ± 10,13 e 4,22 ± 5,19, G-LH). Quanto ao período das ovulações, observou-se grande distribuição nos grupos, sendo possivelmente correlacionado ao grande número de estruturas não fecundadas e embriões degenerados encontrados. A ausência de diferença estatística no número de corpos lúteos, de embriões recuperados e viáveis pode ser associada à grande variabilidade individual. / Abstract: Twenty superovulation programs and embryos recovery were accomplished in Santa Inês sheep, with objective to test the hypothesis that addition of LH at the end of the superovulatory regimen causes an increase in the ovulation rate and of embryos production. For so much, the experiment was delineated in cross-over, being the estrus synchronized with CIDR, for 14 days. On D7, the implant was substituted by a new one, and 37.5 μg of d-cloprostenol were administered. Then, 256 mg of FSHp were administered in 8 decreasing doses, starting on D12. On D14, all females received 200 IU of eCG and 37.5 μg of d-cloprostenol. In control group (G-C), the FSH/LH ratio was constant, while in G-LH 7.5 mg of LH, administered 24 h after implant withdrawal (D15). The laparoscopic inseminations occurred 42 and 48 h after implant withdrawal, concomitantly, the evaluated ovarian structures. On D21, the embryos were surgically collected and evaluated. Overall, the results showed that LH administration at the end of the FSH treatment did not significant increase the ovulation rate and number of viable embryos (78.7% ± 22.72 and 3.77 ± 4.29, respectively, G-C; 84.76 ± 10.14 and 4.22 ± 5.22, G-LH). For the period of the ovulations, a great distribution was observed inside of the groups, being correlated possibly to the great number of unfertilized oocytes and degenerated embryos. The no statistical difference in the number of corpus luteum, embryos recovery and viability can be associated the great individual variability. / Mestre

SOUSA, Solange Damasceno. Proteômica do fluido da Rete Testis de carneiros Morada Nova. 2015. 133 f. : Dissertação (Mestrado) - Universidade Federal do Ceará, Centro de Ciências, Departamento de Zootecnia, Programa de Pós-Graduação em Zootecnia. Fortaleza-CE, 2015. / Submitted by Eric Santiago (erichhcl@gmail.com) on 2016-08-10T12:26:45Z No. of bitstreams: 1 2015_dis_sdsousa.pdf: 3355993 bytes, checksum: 3f9c9ea989725756a91c3d6679b0614a (MD5) / Approved for entry into archive by Nádja Goes (nmoraissoares@gmail.com) on 2016-08-10T15:18:50Z (GMT) No. of bitstreams: 1 2015_dis_sdsousa.pdf: 3355993 bytes, checksum: 3f9c9ea989725756a91c3d6679b0614a (MD5) / Made available in DSpace on 2016-08-10T15:18:50Z (GMT). No. of bitstreams: 1 2015_dis_sdsousa.pdf: 3355993 bytes, checksum: 3f9c9ea989725756a91c3d6679b0614a (MD5) Previous issue date: 2015 / The aim of the study was to identify and characterize the proteins of the rete testis fluid from Morada Nova rams. The testicles, obtained from six slaughtered Morada Nova rams, were immediately dissected. The head of the epididymis was separated to gain access to the efferent ducts. The fluid from the efferent ducts was obtained by testis massage. Thereafter, the fluid was centrifuged to remove cell debris and sperm. Proteins were precipitated with acetone at -20°C and quantified by the Bradford assay. Each sample (400 µg) was focused in strips of 13 cm (pH 4-7) and the second dimension was conducted on SDS-PAGE 15%. The gels were scanned with an ImageScanner II (GE Lifesciences, USA) and analyzed using the PDQuest® version 8.0.1 (Bio-Rad Laboratories, USA). Spots detected after PDQuest analysis of 2-D maps were cut from gels and submitted to trypsin digestion. Proteins were identified using tandem mass spectrometry. Protein information obtained by MASCOT was analyzed using the software tool for searching annotations of proteins (STRAP). Protein-protein interaction networks were obtained from STRING version 9.0 database. In the gels were detected 227 ± 32.1 spots (mean ± SD), where 51% of the proteins were found above 40 kDa, corresponding to 65% of the intensity of all spots detected. Based on gene ontology analysis, the most common biological processes associated with proteins from rete testis fluid were regulation (24.28%) and cellular process (23.27%). Binding (27.42%) and catalytic activity (19.30%) corresponded to the most frequent molecular functions for those proteins. The most intensely expressed proteins were: albumin, clusterin, serotransferrin, immunoglobulin gamma-1 chain and alpha-2-HS-glycoprotein. The rete testis fluid has a large quantity of proteins related to the spermatogenesis. This feature is important in view of the fact that these molecules contribute to the development of the germ cells, as a result of the transport and conversion of substances required to the production of male gametes. / O objetivo do estudo foi identificar e caracterizar as proteínas do fluido da rete testis de carneiros Morada Nova. Os testículos, obtidos de seis carneiros Morada Nova abatidos, foram imediatamente dissecados. A cabeça do epidídimo foi separada para ter acesso aos ductos eferentes. O fluido oriundo dos ductos eferentes foi obtido por massagem do testículo. Posteriormente, o fluido foi centrifugado para remoção dos debris celulares e espermatozoides. As proteínas foram precipitadas com acetona a -20°C e quantificadas pelo método de Bradford. Cada amostra (400 μg) foi focalizada em tiras de 13 cm (pH 4-7) e a segunda dimensão foi realizada em SDS-PAGE a 15%. Os géis obtidos foram escaneados com um ImageScanner II (GE Lifesciences, EUA) e analisados utilizando o aplicativo PDQuest® versão 8.0.1 (Bio-Rad Laboratories, EUA). Os spots detectados após a análise dos mapas 2-D no PDQuest foram cortados dos géis e submetidos a digestão com tripsina. As proteínas foram identificadas por espectrometria de massa em tandem. A informação sobre a proteína adquirida pela busca no MASCOT foi analisada através da utilização do programa para anotações de proteínas (STRAP). Redes de interação proteína-proteína foram obtidas a partir do banco de dados STRING versão 9.0. Nos géis foram detectados em média 227 ± 32,1 spots, onde 51% das proteínas se encontraram acima de 40 kDa representando 65% da intensidade de todos os spots detectados nos géis. Com base na análise da ontologia gênica, os processos biológicos mais comuns associados às proteínas do fluido rete testis foram regulação (24,28%) e processos celulares (23,27%). Ligação (27,42%) e a atividade catalítica (19,30%) corresponderam às funções moleculares mais frequentes. As proteínas mais intensamente expressas foram: albumina, clusterina, serotransferrina, imunoglobulina gama-1 e alfa-2-HS-glicoproteína. A maioria das proteínas identificadas desempenha importantes papéis nos processos de espermatogênese, proteção espermática, motilidade, capacitação e reação acrossômica. O fluido da rete testis possui várias proteínas envolvidas na espermatogênese, o que representa um importante fator, uma vez que estas moléculas contribuem para o desenvolvimento das células germinativas, participando no transporte e conversão de substâncias requeridas para a produção dos gametas masculinos.

A perda anual de ovelhas em criações de sistema extensivo apresenta altos níveis na região Sul do Brasil. Na Serra Gaúcha essa porcentagem chega a 30% e torna-se um fator alarmante para propriedades com sistemas de produção de carne, leite e lã. As perdas reprodutivas em ovinos desta região estão relacionadas com a baixa taxa de concepção do rebanho e alta mortalidade perinatal de cordeiros (MPC) e frequentemente esses fatores são decorrentes de enfermidades, deficiências nutricionais durante o período gestacional e pós-parto ou por manejo inadequado de ovelhas e cordeiros. A MPC, definida como a morte de recém-nascidos imediatamente antes ou durante o parto e até os primeiros vinte e oito dias de vida é uma das principais causas de baixa taxa de desmame no Rio Grande do Sul (RS). Sua etiologia envolve ações complexas que circundam a ação individual e a interação de muitos fatores relacionados entre si. Estudos sobre as causas de MPC no RS têm apontado o complexo exposição/inanição e hipotermia, além da distocia como as duas principais patologias envolvidas nesses óbitos. Condições ambientais adversas, como o frio severo, também causam a morte em consequência da falta de adaptação do recém-nascido às novas condições de vida. A busca de informações que venham a elucidar as causas dessas perdas motivou a realização do presente estudo e o uso da esquila durante a gestação foi utilizado como uma possível ferramenta para minimizar a MPC. O efeito da esquila préparto (74 dias de gestação) sobre sobre o perfil metabólico e produtivo de ovelhas e no peso e desenvolvimento de seus cordeiros durante o primeiro mês de vida foram avaliados neste estudo. Um rebanho de 40 ovelhas gestantes da raça Corriedale foi dividido aleatoriamente em dois grupos: ovelhas com esquila completa (EC) e ovelhas mantidas com velo ou ovelhas controle (OC). As ovelhas e seus respectivos cordeiros foram avaliados em três momentos distintos durante o experimento: no parto, entre 15 e 21 dias de lactação e entre 22 e 45 dias de lactação. Os parâmetros de escore de condição corporal, dosagem de beta-hidroxibutirato, hematócrito, hemoglobina, lactato, glicose, peso corporal, peso da placenta e produção do leite foram mensurados e correlacionados. Os valores médios de hematócrito, hemoglobina e peso dos cordeiros apresentaram diferenças significativas (p< 0,05) entre os grupos EC e OC. O peso médio da placenta e a produção de leite apontaram diferença significativa enquanto que os achados para escore de condição corporal (ECC) e beta-hidroxibutirato não evidenciaram diferenças entre os grupos EC e OC, considerando todos os períodos em conjunto (p> 0,05). A produção leiteira de ovelhas do grupo EC (1.261,25 mL/dia) foi maior (p <0,05) do que no grupo OC (937,79 mL/dia). A esquila de ovelhas aos 74 dias de gestação mostrou-se como importante ferramenta para o melhor desenvolvimento de cordeiros na fase pós-nascimento de forma a contribuir para a diminuição da taxa de mortalidade perinatal. / The annual loss of sheep in extensive system has expressive levels in southern Brazil. It can reach 30% in some areas and becomes an alarming factor for properties with systems producing meat, milk and wool. Reproductive losses in sheep in this region are related to low conception rate of the herd and a high lamb perinatal mortality (LPM). Often these factors are due to diseases, nutritional deficiencies during pregnancy and postpartum period or inadequate management of ewes and lambs. Perinatal mortality is defined as lambs in the death of the neonate until the first twenty-eight days. The LPM is responsible for the low rate of weaning in Rio Grande do Sul (RS), southern Brazil. Its etiology involves complex actions and interaction of many factors related to each other. Studies about causes of the LPM in RS have showed the complex exposure, hypothermia and starvation, in addition to dystocia as the two main pathologies involved in these deaths. Adverse environmental conditions, such as severe cold, cause death due to the lack of adaptation of the newborn to the new living conditions also. The search for more information that might elucidate the causes of these losses motivated the present study and the use of shearing during pregnancy has been used as a possible tool to minimize LPM. The effect of pre-partum shearing (74 days gestation) on the metabolic and productive profile and weight of ewes and their lambs develop during the first month of life were studied. A flock of 40 pregnant Corriedale ewes were randomly divided into 2 groups: ewes completely sheared (EC) and ewes maintained with fleece or sheep control (OC). The ewes and their lambs were evaluated at three different times during the experiment: at birth, between 15 and 21 days of lactation and between 22 and 45 days of lactation. The parameters such as body condition score, betahydroxybutyrate, PVC, hemoglobin, lactate, glucose, body weight, placental weight and milk production were measured and correlated. The mean values of PVC, hemoglobin and lambs weight showed significant differences (p <0.05) between the EC and OC groups. The average weight of placenta and milk production showed a significant difference while the findings for body condition score (ECC) and beta-hydroxybutyrate showed no differences between groups EC and OC, considering all periods together (p> 0.05). Milk production of ewes of group EC (1261.25 mL / day) was higher (p < 0.05) than in the OC group (937.79 mL / day). Shearing of sheep at 74 days of pregnancy was a important tool for the better development of lambs in the post-birth in order to contribute to reducing the rate of perinatal mortality.

Resumo: Avaliou-se a segurança e a eficácia da técnica de biopsia pulmonar transtorácica percutânea e seus efeitos sobre o metabolismo oxidativo na espécie ovina. Foram utilizados 20 ovinos clinicamente sadios e destinados ao abate. Inicialmente, foram coletadas amostras de sangue da veia jugular em tubos de ensaio contendo heparina. Após a colheita, foi realizado o exame clínico seguido da biopsia pulmonar com agulha semi-automática no 7º espaço intercostal direito cerca de 5 cm acima do olécrano. Avaliou-se o número de tentativas para a realização do procedimento e o tamanho dos fragmentos. As amostras obtidas foram analisadas histologicamente. Uma nova colheita de sangue foi realizada 30 minutos após a biopsia, e as amostras levadas ao laboratório para a detecção indireta dos radicais livres [superóxido dismutase (SOD), glutationa total (GSH-t) e peroxidase (GSH-Px) e substâncias reativas ao ácido tiobarbitúrico (TBARS)]. Após o abate foi realizado o exame pos mortem para avaliação de complicações da técnica e das eventuais lesões provocadas. Entre todos os animais submetidos à biopsia apenas dois demonstraram resistência à técnica, sendo obtidos fragmentos pulmonares que mediram entre 4 mm a 7 mm de comprimento, com média de 1,8 1 tentativas por fragmento. As principais alterações relacionadas à técnica foram: tosse, dispnéia inspiratória, elevação das freqüências cardíaca e respiratória e aumento do ruído broncobronquiolar. Ao exame pos mortem observou-se hemorragia dos músculos intercostais e pleura visceral de tamanho variado em todos os cordeiros, além de feridas lineares de até 14 mm nos dois animais que não reagiram bem ao procedimento. Das 20 tentativas de execução da técnica, 18 obtiveram sucesso, enquanto que em duas o fígado foi equivocadamente puncionado. As amostras de tecido pulmonar foram consideradas representativas... (Resumo completo, clicar acesso eletrônico abaixo) / Abstract: It was evaluated the safety and effectiveness of the percutaneous transthoracic lung biopsy technique and its effects of the procedure on oxidative metabolism in sheep. It was used 20 clinically healthy sheep. Initially, blood samples were collected from the jugular vein in test tubes containing heparin. After collecting the blood, the clinical examination was carried out followed by lung biopsy with a semi-automatic needle biopsy. The number of attempts and size of the fragments were measured and the samples were analyzed histologically. New blood samples were collected 30 minutes after the biopsy. All blood samples were taken to the laboratory for the indirect detection of free radicals [superoxide dismutase (SOD), total glutathione (GSH-t), glutathione peroxidase (GSH-Px) and thiobarbituric acid reactive substances (TBARS)]. After the slaughter, a post mortem examination assessed the technical complications and possible injuries. Among all animals underwent biopsy only two did not tolerate the technique. It were obtained Lung fragments from 4 mm to 7 mm and it were required on average 1,8 attempts. The main changes related to the procedure were: cough, labored breathing, increased heart and breathing rate and increased broncobronquiolar noise. The post-mortem examination revealed bleeding of the intercostal muscles and visceral pleura of various sizes in all lambs, and a wound up to 14 mm in the lung of two animals that did not tolerate the procedure. From 20 attempts to implement the technique, 18 were successful, while in two of them the liver was mistakenly punctured. Samples of lung tissue were representative. It was possible to visualize intact structures, including bronchioles and alveoli. The activity of SOD and GSH-Px decreased thirty minutes after the biopsy. There were no changes in GSH-t and TBARS concentrations after the procedure. The percutaneous transthoracic lung... (Complete abstract click electronic access below) / Orientador: Roberto Calderon Gonçalves / Coorientador: Débora Cristina Damasceno / Banca: Simone Biaggio Chiacchio / Banca: Fernando José Benesi / Mestre

Made available in DSpace on 2014-06-11T19:27:15Z (GMT). No. of bitstreams: 0 Previous issue date: 2010-07-02Bitstream added on 2014-06-13T18:55:45Z : No. of bitstreams: 1 barbante_p_me_botfmvz.pdf: 14043998 bytes, checksum: de62a414c23b2ef4f92ffe0263efd2d7 (MD5) / Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP) / Nos últimos anos a ovinocultura reapareceu na região Sudeste, principalmente no estado de São Paulo, como solução econômica para os pecuaristas. Entretanto, havendo um estado sanitário deficiente na criação, pode-se haver a instalação de doenças nos rebanhos e diminuição na produção, como ocorre na leptospirose, uma das zoonoses mais representativas. Com o intuito de estudar a presença de Leptospira spp. nos rebanhos ovinos, foram colhidas 100 amostras sanguíneas e os respectivos fragmentos de fígado e rim dos animais de 20 diferentes propriedades, durante o abate em um frigorífico da região de Sorocaba/SP. Pela prova de Soroaglutinação Microscópica (SAM) obteve-se 23 amostras sorológicas positivas (23%) para um ou mais sorovares de Leptospira spp., com prevalência do sorovar Autumnalis. Para a pesquisa do agente em fragmentos de fígado e rim, 100 amostras de cada tecido foram cultivadas em meio de Fletcher, obtendo-se oito (4%) amostras positivas, sendo quatro para rim e quatro para fígado. Destes, cinco animais apresentaram sorologia positiva (um animal positivo simultaneamente para fígado e rim) e dois negativos. Com a prova de Reação em Cadeia pela Polimerase (PCR) para Leptospira spp. houve positividade em 14 (7%) amostras, sendo sete de rim e sete de fígado. Destes, dez eram animais positivos sorologicamente (dois animais positivos simultaneamente para fígado e rim) e dois negativos. Em relação a técnica de cultivo em meio de Fletcher®, a prova PCR demonstrou-se mais rápida, prática e sensível na detecção de leptospira. Pelos resultados obtidos ressalta-se a importância da espécie ovina no contexto epidemiológico da leptospirose / In recent years, sheep farming has reappeared in the Southeast region. However, having a deficient sanitary state, disease can appear in flocks and production reduce, which occurs with leptospirosis. With the aim of studying Leptospira spp. in sheep herds, 100 blood samples and respective kidney and liver fragments were collected from animals from 20 different properties during slaughter at a meat company in the Sorocaba region, SP, Brazil. The microscopic agglutination test (MAT) found 23 serologically positive samples for one or more Leptospira spp. serovars, with an antileptospire antibody occurrence rate of 23% and more significantly for the Autumnalis serovar. To study the agent in the liver or kidney, 100 samples of each tissue were submitted to culture in Fletcher medium using the Pasteur pipette method and analysed by the Polymerase Chain Reaction test, obtaining 12 positive animals considering both agent detection techniques. The Leptospira spp. cultures presented eight positive samples (four kidney and four liver), corresponding to five animals with positive serology (one animal simultaneously positive for both kidney and liver) and two negatives. PCR detected Leptospira in 14 samples (seven kidney and seven liver) corresponding to 12 positive animals (two animals simultaneously positive for kidney and liver), of these, ten were serologically positive and two negative.PCR was faster, more practical and sensitive than culture for detecting leptospire. Our results stress the importance of ovine specie in the epidemiological context of leptospirosis, acting as disease reservoirs and carriers for the environment and for rural and meat process workers