Dissertations / Theses on the topic 'Porcia'

La Serenissima nel corso dei secoli si è sempre distinta per aver dato i natali ad artisti illustri, che hanno contribuito ad esportare nel mondo l’immagine della città lagunare e dei suoi principali esponenti. Del resto non stupisce che in un contesto sociale in cui l’affermazione personale rappresentava motivo di vanto e prestigio, si vedano fiorire scuole di ritrattisti, tutt’ora tra le più famose al mondo. Se a cavallo tra il XVI e XVII le punte di diamante del ritratto a Venezia erano, un ormai affermato ed anziano Tintoretto e un fiorente Leandro Bassano, nell’entroterra troviamo dei veri e propri alter ego dei maestri lagunari, pur con tutte le ristrettezze proprie dell’arte di provincia. È questo il caso di Francesco Apollodoro detto il Porcìa (1531-1612) che si configura come il ritrattista per eccellenza dell’aristocrazia padovana. Nonostante le fonti storiche a lui coeve lo descrivano come “omo stimato in Padova nel far de’ ritratti”, la critica moderna non è stata altrettanto clemente, in quanto, per scarsità di fonti si è riusciti solo ad avere un’idea frammentaria della sua produzione. In questo lavoro si cercherà di fare luce sulle criticità ed i pregi dell’artista, seguendo un’ipotesi di ordinazione delle opere a lui attribuibili e lo stato dell’arte, al fine di produrre un catalogo ragionato.

As lesões cutâneas e queimaduras são considerados os principais causadores de danos e perdas dos tecidos moles. Em casos severos de trauma, os processos naturais de regeneração são insuficientes no reparo dos danos, resultando em lesões cutâneas crônicas. A desvitalização de matrizes homólogas ou heterólogas é uma alternativa para a produção de matrizes dérmicas. A pele porcina é bastante similar à pele humana, podendo ser utilizada como matriz de colágeno na regeneração de tecido mole. Além disso, ela tem como constituinte principal o colágeno tipo I, e, assim, pode ser utilizada em queimaduras de segundo grau. Este trabalho teve como objetivo a preparação e caracterização de matrizes extracelulares de colágeno tipo I por meio de hidrólise alcalina e reticulação com glutaraldeído (GA). As matrizes de colágeno foram obtidas a partir da hidrólise alcalina de pele porcina, com posterior reticulação com GA, em diferentes concentrações (0-0,1%) e tempos de reação (15 e 45 min). As matrizes foram caracterizadas através de determinação do conteúdo de elastina, estabilidade biológica (tripsina), Calorimetria Exploratória Diferencial (DSC), termogravimetria (TG/DTG), Microscopia Eletrônica de Varredura (MEV) e citotoxicidade in vitro. Através da determinação do conteúdo de elastina, foi possível determinar a massa média de colágeno presente nas matrizes, a qual foi de 95,2?0,2% (m/m), e a massa média de elastina, que foi de 4,8?0,2% (m/m), e também verificar que independente do tratamento, a elastina estava presente nas matrizes. O ensaio de estabilidade biológica mostrou que o tratamento com GA diminui a biodegradação do material; sendo obtidos porcentagens de degradação que variaram de 83,6%?1,1 (0% GA) a 46,1%?0,7 (0,085%-45min), indicando, assim, que com o aumento da concentração de GA e do tempo de reação, há uma diminuição da porcentagem de degradação. Pela análise termogravimétrica, foi observado que o colágeno presente nas matrizes tornou-se mais estável termicamente em conseqüência do aumento do grau de reticulação e, portanto, mais resistentes à degradação térmica. Os resultados de DSC confirmam os de termogravimetria devido ao aumento nos valores das temperaturas de desnaturação das matrizes em função do aumento do tempo de reação e da concentração de GA. Pela análise das fotomicrografias, pôde ser observado que após a reticulação com GA, as fibras de colágeno tornam-se mais organizadas e definidas; e essa definição torna-se maior com o aumento da concentração de GA. Os resultados de citotoxicidade in vitro mostraram que as matrizes analisadas são citotóxicas possivelmente devido a gordura remanescente, sendo necessário a realização de um pré-tratamento. Assim, a preparação de matrizes derivadas de pele porcina com diferentes tempos de degradação, as quais podem ser utilizadas na reconstrução de tecidos moles, é viável.
Cutaneous lesions and burns are considered the main causes of damage of soft tissues. In severe cases of trauma, the natural processes of regeneration are insufficient in the repair of the damage, resulting in chronic cutaneous lesions. Desvitalization of homologous or heterologous matrices is an alternative for the production of dermal matrices. The porcine skin is quite similar to the human skin and can be used as collagen matrix in soft tissue regeneration. Besides, it contains type I collagen as the main constituent and thus, it can be used in second degree burns. The objective of this work was the preparation and characterization of type I collagen extracellular matrices with alkaline hydrolysis and glutaraldehyde (GA) crosslinking. The collagen matrices were obtained from the alkaline hydrolysis of porcine skin, with subsequent GA crosslinking, in different concentrations (0 - 0,1%) and reaction time (15 and 45 min). Matrices were characterized by determination of the elastin content, biological stability (trypsin), differential scanning calorimetry (DSC), termogravimetry (TG/DTG), scanning electron microscopy (SEM) and a preliminar assay of in vitro cytotoxicity. Elastin and collagen content were 4,8±0,2% (m/m) and 95,2±0,2% (m/m), respectively. Biological stability results showed that GA crosslinking reduces matrix biodegradation; as degradation varied from 83,6%±1,1 (0% GA) to 46,1%±0,7 (0,085% - 45min), demonstrating, thus, that with the increase of GA concentration and reaction time, there was a decrease of degradation. For termogravimetric analysis it was observed that the collagen present in the matrices become termically more resistant as a consequence of the increasing crosslink degree and, therefore, more resistant to thermal degradation. DSC results, similar to termogravimetric ones, showed an increase in denaturation temperatures as a function of increasing reaction time and GA concentration. SEM analysis showed that after the GA crosslinking, collagen fibers become more organized and defined; and that definition improved with increasing GA concentration. Preliminar assay of in vitro cytotoxicity showed that treated matrices are cytotoxic possibly due to remaining fat, being necessary the accomplishment of a pre-treatment. Therefore, porcine skin matrices preparation with different degradation times, which can be used in the soft tissue reconstruction, are viable.

A potencialidade do uso de uma matriz de colágeno e de um creme preparado pela mistura de um creme hidratante comercial com um gel de colágeno (90:10 m/m) foi avaliada no processo de cicatrização de feridas planas produzidas em pele de ratos. A matriz acelular de colágeno e o gel de colágeno foram obtidos a partir de serosa porcina por meio de um tratamento alcalino que não altera a estrutura do colágeno nativo e remove componentes celulares. Este estudo fez um comparativo macroscópico e histológico do processo cicatricial das feridas planas tratadas com soro fisiológico ou creme comercial (controles) e as tratadas com a sutura de uma membrana de colágeno (matriz) ou creme com colágeno. As feridas foram produzidas pela retirada de um flap de pele de 20 'MM POT.2' e receberam curativos diários. O material para histologia foi coletado nos 3º, 5º, 7º e 9º dias pós-cirurgia. Apesar de não ter havido uma acentuada diferença na cicatrização das feridas planas dos dois grupos de controle e no grupo que recebeu tratamento do creme contendo colágeno, a presença deste no creme indicou uma pequena diferença no grau de colageinização, o que demonstra serem válidas mais investigações nesta direção, buscando uma melhor proporção creme:gel e/ou diferentes concentrações para o gel de colágeno. A membrana demonstrou ser uma ótima opção para o reparo de lesões por ser de fácil obtenção e armazenamento, ter baixo custo e ser excelente para manuseio (maleável e resistente), além de atender às principais exigências mencionadas na literatura para qualquer curativo biológico oclusivo
The potentiality of the use of a collagen based matrix and of a cream prepared by the mixture of a commercial cream plus collagen (90:10 w/w) were evaluated in the healing process of rats’ skin. The acellular collagen matrix and the collagen gel were obtained by an alkaline treatment of porcine serosa which does not damage the native collagen structure and removes cellular components. This study compared by macroscopy analysis and histology the skin healing repair of wounds treated with physiological solution or commercial cream (control groups) and those treated with collagen based matrix suture or commercial cream plus collagen mix. The wounds were made by removing a skin flap with 20 'MM POT.2' and have received treatment every day. The material for histology was retired on the 3rd, 5th, 7th and 9th days after surgery. Even without an accentuated difference in the healing process of both control groups and the wounds treated with the commercial cream plus collagen, its presence in the cream showed a small difference of the collagen level in the new skin what validate more investigations on this way, searching better cream:gel proportion and/or different gel concentration. The matrix demonstrated to be a very good option to help wound healing because it is easily shelf able and obtainable, it has cheap cost and it is extremely nice to handle (resistant and manipulation able), besides to follow the main requirements present in the literature citation for any biologic occlusive dressing

El objetivo del estudio fue evaluar la persistencia de los anticuerpos pasivos contra el virus de la Peste Porcina Clásica (vPPC) en lechones de dos granjas tecnificadas A y B con distintos programas de vacunación contra el vPPC. En la granja A, las marranas son vacunadas a los 90 días de gestación y en B a los 18 a 21 días post-parto. Se colectaron 60 muestras de sangre de lechones por granja, durante la primera (n igual a 15), tercera (n igual a 15), quinta (n igual a 15) y séptima (n igual a 15) semana de edad, así como de las marranas (n igual a 15) de cada granjas para la detección de los anticuerpos mediante la prueba de ELISA indirecta o de cloqueo. En la primera semana de edad el 100% de lechones de ambas granjas presentaron anticuerpos pasivos, persistiendo dichos anticuerpos en la mayoría de lechones por encima de séptima semana de edad. Se detectaron diferencias en los niveles de anticuerpos pasivos en los lechones durante la primera y tercera semana de edad de ambas granjas siendo estadísticamente significativa (p menor a 0.05). Al comparar los coeficientes de variación de los resultados de los lechones de ambas granjas, se observó una mayor variabilidad en los niveles de anticuerpos en lechones de la granja A. Así mismo, hubo variación en los niveles de anticuerpos en las marranas de la granja A en comparación a los resultados de las marranas de la granja B pero, esta variación no tuvo significancia estadística (p mayor a 0.05). Estos resultados sugieren que los niveles y la persistencia de los anticuerpos pasivos dependen del sistema de manejo de cada granja. Palabras Claves: virus de la Peste Porcina Clásica (vPPC), anticuerpos maternales, lechones, marranas, vacunación, granjas porcinas.
--- The persistence of the levels of maternal antibodies against the of the Classical Swine Fever virus (CSFV) in piglets born of vaccinated sows from two farms (A and B) with different vaccination programs against CSFV located in Lima valley, Peru, was evaluated. In the farm A, the sows are vaccinated at 90 days of gestation and in farm B at 18 - 21 days post furrowed. Serum samples were taken from a total of 60 piglets by farm, at first (n is equal to 15), third (n is equal to15), fifth (n is equal to 15) and seventh (n is equal to 15) weeks old and from sows from farm A (n is equal to 15) and B (n is equal to15) for antibodies detection against CSFV by indirect ELISA test. The 100% (30/30) of piglets of both farms had maternal antibodies against CSFV at first week old. In the majority of piglets the maternal antibodies persisted up to seventh week old. The levels of maternal antibodies in the piglets from both farms showed a statistically significant (p menor a 0,05) differences at first and third week old. The comparison of the maternal antibodies titers indicated more variation in piglets from farm A, a high and uniform antibodies titers were observed in piglets from farm B during the study. The sows had high level of antibodies against CSFV indicating a good passage of these antibodies to their piglets. These results suggest that the level and persistence of the maternal antibodies in the piglets depend of the management system of each pig farms. Key Words: Classical Swine Fever Virus (CSFV), maternal antibodies, piglet, sows, vaccination, pig farms.
Tesis

Memoria para optar al Título Profesional de Médico Veterinario.
El circovirus porcino tipo 2 (PCV-2) es un virus muy pequeño (17 nm. de diámetro), de estructura icosaédrica y con ADN circular simple. Se conoce ampliamente por ser el agente causal de una compleja lista de enfermedades multifactoriales denominadas PCVAD (del inglés, Porcine circovirus associated diseases). Entre estas, una de las más importantes en términos económicos y sanitarios para la industria porcina intensiva, es el PMWS (del inglés, Postweaning multisystemic wasting syndrome), que se caracteriza clínicamente por adelgazamiento progresivo, retraso del crecimiento y desmedro. La infección por si sola por PCV-2 es necesaria, pero no suficiente para desencadenar el cuadro clínico, existen otros factores, tanto individuales como prediales, que participan en el desarrollo del síndrome. Por lo tanto, es más común encontrar individuos infectados con una presentación subclínica, inmunocomprometidos y mayormente susceptibles a coinfecciones. En el presente estudio, se caracterizan los perfiles serológicos de 11 granjas de producción porcina, cuya ubicación geográfica es representativa de la producción a nivel nacional. Un perfil serológico es un estudio efectuado a grupos de animales de diferentes edades o etapas productivas, para detectar la presencia de anticuerpos contra algún patógeno en particular y determinar patrones de infección. De cada granja se tomaron 16 muestras de suero en 3 edades (3-4, 10 y 18-20 semanas). Cada muestra fue evaluada mediante una prueba de ELISA para la detección de anticuerpos anti PCV-2. A modo complementario, los títulos de anticuerpos a las 3 semanas fueron transformados a valores de IPMA (técnica de inmunoperoxidasa en monocapa de cultivo celular), a modo de evidenciar posible interferencia materna con la vacunación. Finalmente, se realizó una recopilación de información respecto a las granjas muestreadas y mediante un Análisis de Varianza (ANOVA) en el programa estadístico Infostat, los factores: status PRRS, vacunación, sistema y tamaño fueron analizados, a modo de establecer posibles diferencias significativas entre las granjas al considerar estos factores. Los resultados muestran un comportamiento característico de los perfiles serológicos, con un alto título de anticuerpos a las 3 semanas, los cuales disminuyen drásticamente en el tiempo, lo que sugiere un eficiente traspaso de inmunidad materna, pero se cuestiona la efectividad en la generación de anticuerpos a través de la vacunación. Además, se observa una alta presencia del fenómeno de interferencia de la inmunidad materna con la vacunación. Por último, los resultados del análisis estadístico muestran que todos los factores analizados generan diferencias significativas entre los perfiles serológicos de las granjas. El estudio permite concluir robustamente que existe evidencia serológica de PCV-2 en todas las granjas analizadas y que existen diferencias estadísticamente significativas entre ellas al considerar ciertos factores prediales, aceptándose la hipótesis de este estudio
Porcine circovirus type 2 (PCV-2) is a very small virus (17 nm of diameter), of icosahedral structure and simple circular DNA. PCV-2 is widely known to be the causative agent of several multifactorial diseases named PCVAD (Porcine circovirus asociated diseases). The most economic important disease caused by PCV-2 is the Postweaning multisystemic wasting syndrome (PMWS), which is characterized by progressive emaciation, growth retardation and wasting. PCV-2 infection is necessary but not sufficient to trigger the clinical presentation, there are other factors, individual and predial, that are involved in the development of the syndrome. Therefore, it is more common to find individuals infected with a subclinical presentation, immunocompromised and most susceptible to coinfections. In the present study, the serological profiles of 11 pig farms were characterized, whose geographic location is representative of production at the national level. A serological profile is a study carried out on groups of animals at different ages, to detect the presence of antibodies against PCV-2, and to determine patterns of infection. From each farm, sera was collected in 3-4 weeks; 10 weeks and 18-20 weeks old pigs, obtaining 16 samples per age. Each sample was evaluated by a comercial PCV-2 ELISA test for antibody detection. Additionally, antibody titres at 3 weeks were transformed to IPMA equivalent values (inmunoperoxidase monolayer assay), in order to evidence possible maternal interference with vaccination. Also, information about farms characteristics and management such as; PRRS status, vaccination, type of production system and size, were collected. All variables were statistically analyzed in order to establish if there were significant differences between the farms when considering these factors. The results show a characteristic behavior of the serological profiles, with a high antibody titre in 3 weeks old pigs, which decrease drastically over time, suggesting an efficient transfer of maternal immunity, but the effectiveness of the vaccination to generate antibodies is questioned. In addition, there is a high presence of the phenomenon of interference of maternal immunity with vaccination. Finally, the results of the statistical analysis show that all the analyzed factors generate significant differences between the serological profiles of the farms. The study allows us to conclude robustly that there is serological evidence of PCV-2 in all farms analyzed and that there are statistically significant differences between them when considering certain property factors, accepting the hypothesis of this study
Financiamiento: Proyecto Zoetis-Favet PCV2.

La present tesi doctoral tenia com a objectiu complementar el coneixement actual sobre l’eficàcia de la vacuna enfront a PCV2 en condicions d’infecció subclínica i explorar dos conceptes nous (en quan a estratègies de vacunació enfront a aquest patogen) que poguessin augmentar l’eficàcia d’aquest tractament. El primer estudi pretenia avaluar la possible interferència de la presència de diferents nivells d’anticossos d’origen maten (AOM) en el moment de la vacunació, en l’evolució del guany mig diari de pes (GMDP). En aquest estudi, només es va detectar una possible interferència en l’eficàcia de la vacuna sobre el GMDP, quan es va considerar la subpoblació d’animals amb els valors S/P més alts. Per tant, l’impacte d’aquesta possible interferència en condicions de camp es probablement negligible en la majoria d’animals i de les granges. En el segon estudi, es va avaluar la viabilitat d’eradicar la infecció de PCV2 en una granja convencional infectada subclínicament amb PCV2 mitjançant una estratègia de vacunació en massa. L’aplicació durant una any de la vacunació en massa enfront a PCV2 (sense implementar mesures específiques de maneig o de bioseguretat) no va ser capaç d’eliminar l’infecció per PCV2. De fet, un cop la vacunació es va aturar, el virus es va detectar de nou. De totes maneres, la disminució dels nivells d’anticossos i la no detecció del virus durant la segona meitat del període de vacunació en massa deixa entreveure que la eradicació de la infecció de PCV2 mitjançant un programa de vacunació més llarg i més extensiu podria ser possible.
This thesis aimed to complement the current knowledge on PCV2 vaccination efficacy under subclinical infection conditions and give new creative concepts (“thinking out of the box”) for future related studies. The first study had the objective to assess the putative interference of different maternally derived antibody (MDA) levels at the time of vaccination on the average daily weight gain (ADWG) evolution. In this study, an apparent interference of vaccine efficacy on ADWG was noticed only when a small subpopulation of pigs with the highest ELISA S/P ratios was considered, Therefore, the impact of this possible interference under field conditions is probably negligible for most of the animals and farms. In the second study, the feasibility to eradicate PCV2 in a conventional PCV2 infected farm by using a mass vaccination strategy was assessed.. One year period of mass PCV2 vaccination (without implementing further farm management practices or biosafety measures) was not able to clear out PCV2 infection. Indeed the virus became detectable again when vaccination was stopped. However, the decreasing antibody levels and the lack of viral detection during the second half of the vaccination period shed a light on eradicating this virus by applying a longer term vaccination in a wider area would be feasible.

A utilização e o desenvolvimento de biomateriais para a regeneração tecidual são de grande importância, principalmente para a área médica e odontológica. Matrizes de colágeno derivadas de tecidos de origem animal são utilizadas devido o colágeno apresentar características como biodegradabilidade e biocompatibilidade. Essas matrizes podem ser obtidas a partir de várias fontes, sendo uma delas o pericárdio porcino, que apresenta vantagens como grande disponibilidade, baixo custo, fácil obtenção e possibilidade de sofrer modificações químicas. Além disso, tecidos de origem suína são muito similares aos tecidos humanos, podendo ser utilizados para a produção de biomateriais para a regeneração de tecido mole. Este trabalho teve como objetivo a preparação de membranas acelulares derivadas de pericárdio porcino por hidrólise alcalina em diferentes tempos, para posterior utilização em engenharia de tecido. As membranas de colágeno foram obtidas por hidrólise alcalina de pericárdio porcino durante 4, 8, 12 e 24 h e caracterizadas por análise histológica, microscopia eletrônica de varredura (MEV), avaliação da citoxicidade in vitro, estabilidade biológica in vitro (colagenase), titulação potenciométrica, porcentagem de absorção de água, calorimetria exploratória diferencial (DSC), termogravimetria (TG), análise térmica dinâmico-mecânica (DMTA) e ensaios de tração. A análise histológica mostrou que após 4h de hidrólise as células foram removidas das membranas. A avaliação da citotoxicidade in vitro mostrou que as membranas preparadas neste trabalho não são citotóxicas. Os ensaios de estabilidade biológica in vitro por colagenase mostraram que as membranas hidrolisadas degradaram mais rapidamente que a não hidrolisada e, quando comparadas com matrizes derivadas de pericárdio bovino, as derivadas de pericárdio porcino foram mais resistentes à degradação por colagenase. A titulação potenciométrica possibilitou determinar o número de grupos carboxílicos das membranas e o incremento desses grupos por molécula de colágeno. Os resultados mostraram que houve um aumento no número de grupos carboxílicos tituláveis nas membranas hidrolisadas e, consequentemente, houve um aumento do número de cargas negativas incorporadas na molécula de colágeno. As membranas hidrolisadas apresentaram uma maior absorção de água, uma diminuição das temperaturas de desnaturaçãoe e menor estabilidade térmica em função do aumento do tempo de hidrólise. Os ensaios de tração mostraram que após a hidrólise alcalina as membranas apresentaram maiores valores de resistência à tração e que a deformação é dependente do tempo de hidrólise alcalina. Esses resultados mostraram que a preparação de membranas de colágeno derivadas de pericárdio porcino com diferentes tempos de hidrólise alcalina é um procedimento viável para ser utilizado na produção de biomateriais para engenharia de tecido.
The use and development of biomaterials for tissue regeneration are of great importance, especially for medical and dental care. Collagen matrices derived from animal tissues are widely used because collagen has characteristics such as biodegradability and biocompatibility. These matrices can be obtained from various sources, such as porcine pericardium, which is a tissue that can be used due its low cost, wide availability and because it can be chemically modified. Besides, porcine tissues are very similar to human tissue and can be used to produce biomaterials for soft tissue regeneration. The aim of this study was the preparation and characterization acellular membranes by alkaline hydrolysis of porcine pericardium. Membranes were characterized by histological analysis, scanning electron microscopy (SEM), in vitro cytotoxicity evaluation, in vitro biological stability (collagenase), potentiometric titration, water absorption percentage, differential scanning calorimetry (DSC), thermogravimetry (TG), dynamic mechanical thermal analysis (DMTA) and tensile tests. Histological analysis showed that after 4h of hydrolysis, cells were totally removed from matrices. In vitro cytotoxicity showed that all matrices prepared in this work are not cytotoxic. In vitro biological stability tests (collagenase) showed that the hydrolyzed membranes degraded more quickly than the non hydrolized matrix and more resistant to collagenase degradation when compared to matrices derived from bovine pericardium. The potentiometric titration allowed the determination carboxylic groups and the increase of these groups per collagen molecule. Hydrolyzed matrices had an increase in water absorption, a decrease in denaturation temperature and a small decrease in thermal stability with the increase of hydrolysis time. Tensile tests showed that after alkaline hydrolysis matrices showed higher tensile strength and the deformation was independent of the time of alkaline hydrolysis. These results showed that the preparation of collagen biological matrices derived from porcine pericardium with different times of alkaline hydrolysis is a viable procedure to be subsequently used in the production of biomaterials for tissue engineering.

El Circovirus porcino 3 (PCV-3) es un virus descubierto recientemente en cerdos domésticos y jabalíes. El virus fue hallado por primera vez en 2016 mediante estudios metagenómicos, en animales afectados por fallo reproductivo, e inflamación cardíaca y multisistémica. Desde entonces, el virus se ha descrito circulando de forma generalizada en animales con diferentes presentaciones clínico/patológicas como en cerdos sanos. Por lo tanto, el objetivo principal de esta Tesis fue generar nueva información sobre la epidemiología molecular del PCV-3 en muestras de cerdos domésticos y jabalíes en España. En el primer estudio, la presencia de PCV-3 en la población porcina española se evaluó retrospectivamente de 1996 a 2017 en sueros de animales de diferentes fases de producción y condiciones clínico/patológicas. La detección del genoma de PCV-3 en estas muestras se realizó mediante PCR y secuenciación del genoma. Los datos obtenidos confirmaron que PCV-3 ha estado circulando en la población porcina española desde el año 1996. La frecuencia global de muestras PCR positivas para PCV-3 en el período de estudio fue 11.47% (75 de 654). El análisis filogenético de las secuencias obtenidas de PCV-3 mostró una alta identidad con las secuencias de PCV-3 ya conocidas. Aunque la información obtenida fue limitada, la presencia de PCV-3 no pareció estar relacionada con ninguna condición patológica específica ni asociada a ninguna fase de producción del cerdo. En el segundo estudio se evaluó la dinámica de la infección por PCV-3. Para ello se analizaron mediante PCR los sueros de 152 cerdos de 4 granjas de alto estatus sanitario y sin problemas clínicos. Los animales fueron monitorizados longitudinalmente 5-6 veces desde las 2 a 4 semanas de edad hasta el final de la fase de engorde. El genoma del PCV-3 se detectó en cerdos de todas las edades y granjas evaluadas; algunos animales presentaron una aparente infección a largo plazo durante un período que varió de 4 a 23 semanas. El análisis filogenético mostró una gran similitud entre las secuencias obtenidas y los genomas de PCV-3 de diferentes países disponibles en las bases de datos. Los resultados confirman que PCV-3 circuló en las granjas estudiadas en España, lo que sugiere que la infección probablemente sea generalizada en el país. La mayoría de los cerdos se infectaron durante su vida productiva, aunque no se encontró asociación con una edad específica. En el tercer estudio, se verificó la frecuencia retrospectiva de la infección por PCV-3 entre 2004 y 2018, así como en una población española de jabalíes capturados y recapturados. Los resultados obtenidos confirmaron la susceptibilidad del jabalí a la infección por el virus, mostrando alta frecuencia de detección de PCV-3 (221 de 518, 42.66%) y demostrando circulación al menos desde el año 2004. Los datos compilados sugieren que PCV-3 es aparentemente capaz de causar una infección persistente, ya que 5 de 10 jabalíes capturados/re-capturados positivos a PCV-3 mostraron positividad en muestreos separados por más de 5 meses. La frecuencia de detección del genoma de PCV-3 también fue investigada por primera vez en diferentes muestras de tejido y heces. Se detectó el genoma de PCV-3 en todos los tipos de tejido analizados. La cantidad de ADN en todas las muestras de PCR positivas para PCV-3 analizadas fue de moderada a baja. Todas las secuencias parciales y completas de PCV-3 obtenidas de jabalíes mostraron una elevada similitud nucleotídica (> 98%).  En conclusión, los resultados obtenidos en esta Tesis proporcionan datos relevantes sobre la epidemiología de este nuevo virus, PCV-3, tanto en cerdos domésticos como en jabalíes. Además, la información filogenética sugiere una baja variabilidad genética de PCV-3, en contraste con otros virus de ADN monocatenario.
Porcine circovirus 3 (PCV-3) is a recently discovered circovirus species found in domestic pigs and wild boar. The virus was found in 2016, through metagenomic sequencing approach, in animals affected by reproductive failure, cardiac and multisystemic inflammation. Since then, the virus has been described in pigs with different clinical/pathological presentations as well as in healthy ones, with a widespread circulation. Therefore, the main objective of this Thesis was to gain insights into the molecular epidemiology of PCV-3 in samples from domestic pigs and wild boar from Spain. In the first study, the presence of PCV-3 in the Spanish pig population was retrospectively evaluated from 1996 to 2017 in sera from animals of different production phases and clinical/pathological conditions. The detection of PCV-3 genome in such samples was attempted by PCR and partial genome sequences were obtained from selected PCV-3 positive samples from different years. Compiled data confirmed that PCV-3 has been circulating in the Spanish pig population since 1996. The overall frequency of PCV-3 PCR positive samples in the study period was 11.47% (75 out of 654). Phylogenetic analysis of the PCV-3 obtained sequences showed high identity with the already known PCV-3 sequences, with low variations among years. Although the available information was limited, PCV-3 did not appear to be linked to any specific pathological condition or pig age-group. The second study aimed to assess the dynamics of PCV-3 infection by means of PCR in serum. A total of 152 pigs from 4 different healthy farms, which were sampled longitudinally five or six times from 2-4 weeks of age until the end of the fattening period, were analyzed. PCV-3 genome was found in pigs from all tested ages and farms; few animals had an apparent long-term infection during a period ranging from 4 to 23 weeks. Phylogenetic analysis showed high similarity among the obtained sequences and with available PCV-3 genomes from different countries. Results confirmed that PCV-3 circulated in all studied farms from Spain, suggesting that infection is probably widespread in the country. Most pigs got infection during their life, although PCV-3 did not appear to circulate mostly at any specific age. In the third study, the frequency of PCV-3 infection was retrospectively assessed in Spanish wild boar from 2004 to 2018, as well as in captured and re-captured animals. Obtained results confirmed the susceptibility of wild boar to the virus, showing high frequency of PCV-3 detection (221 out of 518, 42.66%) and demonstrating circulation at least since 2004. Compiled data suggests that PCV-3 is apparently able to cause persistent infection, since 5 out of 10 PCV-3 PCR positive captured/re-captured boars showed positivity in samplings separated for more than 5 months. The frequency of PCV-3 genome was also investigated for the first time in different tissue samples and feces, where all tested tissue types’ harbored PCV-3 genome. The amount of DNA in all tested PCV-3 PCR positive samples was moderate to low. All partial and complete PCV-3 sequences obtained from wild boar displayed high nucleotide similarity (>98%). In conclusion, the obtained results of this Thesis provide relevant data on the epidemiology of this novel virus, in both domestic pig and wild boar, which appear to be widespread. Moreover, the phylogenetic information suggests low genetic variability of PCV-3, in contrast with other single stranded-DNA viruses.

En les últimes dècades, el sector porcí ha experimentat un fort desenvolupament caracteritzat per un augment en la producció, en el cens, en les exportacions, així com en la productivitat de les seves explotacions. L'objectiu principal de la present investigació és identificar factors que impulsen la rendibilitat econòmica de les empreses dedicades a la producció de bestiar porcí, els quals permetin millorar les decisions de gestió i localització. Aquests factors poden ser intrínsecs a l'empresa, sectorials i referits a nivell territorial i geogràfic. Les dades utilitzades en l'estudi provenen d'una mostra de 1.810 empreses espanyoles que proporcionen un conjunt de dades de panell no balancejat per al període 2003-2018, el tractament estadístic s'ha dut a terme mitjançant l'aplicació d'eines estadístiques per a dades de panell, que han permès la detecció dels factors més influents en la rendibilitat de les empreses tenint en compte la possible existència de problemes d'endogeneïtat entre algunes de les variables analitzades. Els resultats permeten deduir implicacions en dos sentits; d'una banda, per als directius existents amb motiu de millorar l'acompliment empresarial i, d'altra banda, per a futurs inversors que pretenguin emprendre noves inversions en el sector. Aquests resultats poden ser d'utilitat per al negoci, ja que revelen com les empreses poden obtenir majors rendiments econòmics.
En las últimas décadas, el sector porcino ha experimentado un fuerte desarrollo caracterizado por un aumento en la producción, en el censo, en las exportaciones, así como en la productividad de sus explotaciones. El objetivo principal de la presente investigación es identificar factores que impulsan la rentabilidad económica de las empresas dedicadas a la producción de ganado porcino, los cuales permitan mejorar las decisiones de gestión y localización. Estos factores pueden ser intrínsecos a la empresa, sectoriales y referidos a nivel territorial y geográfico. Los datos utilizados en el estudio provienen de una muestra de 1.810 empresas españolas que proporcionan un conjunto de datos de panel no balanceado para el período 2003-2018, cuyo tratamiento estadístico se ha llevado a cabo mediante la aplicación de herramientas estadísticas para datos de panel, que han permitido la detección de los factores más influyentes en la rentabilidad de las empresas teniendo en cuenta la posible existencia de problemas de endogeneidad entre algunas de las variables analizadas. Los resultados permiten deducir implicaciones en dos sentidos; por un lado, para los directivos existentes con motivo de mejorar el desempeño empresarial y, por otro lado, para futuros inversores que pretendan acometer nuevas inversiones en el sector. Estos resultados pueden ser de utilidad para el negocio, ya que desvelan cómo las empresas pueden obtener mayores rendimientos económicos.
In the last decades, swine sector has experienced a strong development characterized by an increase in production, census, exports as well as in productivity. For this reason, the main objective of this research is to identify factors that drive the profitability of companies dedicated to the production of pigs which allow them to improve their management and location decisions. These factors can be intrinsic to the company, sectorial and referred to territorial and geographical level. The data used in this study come from a sample of 1,810 spanish companies that provide an unbalanced panel data set for the period 2003-2018, whose statistical treatment has been carried out through the application of frequent statistical tools. As expected, there are certain factors that are decisive in the profitability of companies, so the results yield relevant conclusions in two ways; on the one hand, for existing managers in order to improve business performance and, on the other hand, for future investors who intend to undertake new investments in the sector. These results are important because they have practical implications for the business.

La presente tesis plantea un modelo de sistema experto que permita realizar la identificación del virus de la peste porcina clásica. Se considera como objeto de estudio el diagnostico de la peste porcina clásica que pertenece al conjunto de enfermedades víricas. Este tipo de afección se presenta con frecuencia dentro de una explotación por la mala práctica de la sanidad porcina. Se diseña el sistema experto que cuente con las siguientes características: los síntomas del paciente consideradas variables de entrada, una base de hechos que almacene síntomas particulares, una base de conocimientos diseñado con lógica de predicados formalizadas que contienen el conocimiento del experto veterinario, un motor de inferencia que obtiene conclusiones partir de hechos y reglas por medio del encadenamiento hacia atrás, y finalmente la variables de salida (diagnostico). Con la ayuda de un sistema experto diferentes personas con poca experiencia pueden resolver problemas en lugar de personas que requieran un conocimiento mas especializado. Esto se debe a que el sistema experto pueden obtener conclusiones y resolver problemas de forma mas rápida que los expertos humanos ya que el razonamiento humano esta en base a un conocimiento adquirido. Posteriormente con los datos obtenidos de las pruebas, se evalúa el sistema experto llegándose a la conclusión que los diagnósticos obtenidos tienen un grado de confiabilidad de un 91% respecto a los resultados reales

La cisticercosis porcina es una enfermedad parasitaria de carácter zoonótico. Los estudios de cisticercosis porcina en nuestro país se han realizado en diferentes puntos geográficos, obteniéndose prevalencias altas. Esta última característica ha calificado al Perú como área endémica. Sin embargo, no existe ningún trabajo de investigación para esta enfermedad en la costa central, lugar donde se desarrollan centros de crianza informal de cerdos que aparentemente podrían ser focos difusores de la enfermedad. El objetivo del presente estudio fue determinar la prevalencia de la cisticercosis porcina en el área de ampliación del Parque Porcino: “Pampas de los Perros” que se divide en cinco sectores (A, B, C, D y E), y se localiza en el Distrito de Ventanilla, Provincia Constitucional del Callao. Se muestraron un total de 299 cerdos de los cuales se encontraron 55 animales positivos a la presencia de anticuerpos mediante la prueba de EITB. La seroprevalencia a la prueba fue 18 + 4% y la seroprevalencia corregida de 19 + 4%. No se encontró diferencia estadística significativa entre los sectores de procedencia ni el sexo. Se halló diferencia significativa (p<0.05) entre el grupo etáreo mayor de 12 meses y los otros tres grupos (<4, 4-8 y de 8-12 meses). Mediante la prueba de regresión logística el grupo (> 12 meses) y los sectores A y B, fueron calificados como factores de riesgo para la presentación de la cisticercosis (p<0.05). Se empleó la prueba de simulación estocástica beta-binomial para obtener un valor más confiable de la presentación de la enfermedad, determinándose una prevalencia real de 7% y que el 90% de las observaciones se encontraría en el intervalo de 4% a 10%.
Porcine cysticercosis is a parasitic zoonotic disease. Studies in Peru have been conducted in several different geographic areas, where prevalence rates are high. Consequently, Peru has been declared an endemic country. So far, no studies into porcine cysticercosis have been undertaken on the central coast. In this region non intensive pig farming exists, which may generate a focal point for disease transmission. The aim of this study was to determine the prevalence of porcine cysticercosis in the porcine breeding area called “Parque Porcino Pampas de los Perros” in the Ventanilla district, Callao Province, Peru. This Porcine breeding area is divided into 5 sections; A, B, C, D and E. A total of 299 pigs were sampled, from which 55 were positive to EITB test. Seroprevalence results from the test were 18 ± 4%. There was no statistically significant difference between gender of the pigs or section (A B C D or E). Statistically significant difference was found in the age group greater than 12 months old (p <0.05). Logistical regression tests revealed that the age group> 12 months old and the sections A and B are risk factors for contracting the disease (p<0.05). A beta binomial stochastic simulation test was applied to obtain reliable values for disease interpretation, obtaining a real prevalence of 7%. This demonstrated that 90% of observations were recorded between the 4% and 10% interval.
Tesis

En els darrers anys, l’ús dels fluids orals (FO) com a mostra diagnòstica en el control i monitoratge del Virus de la síndrome reproductiva i respiratòria porcina (PRRSV) s’ha estès en gran mesura per l’Amèrica del Nord i de manera més discreta per Europa. Tanmateix, no existeixen gaires estudis sobre els límits de detecció de la tècnica en el monitoratge de la malaltia en granges infectades amb el PRRSV1. Per aquesta raó l’objectiu de la present tesi va ser avaluar l’eficiència de l’ús dels fluids orals en el diagnòstic del PRRSV1, determinant els factors que influeixen en la seva eficàcia diagnòstica. L’objectiu del primer estudi va ser l’avaluació de l’excreció individual en FO del PRRSV1 al llarg del temps, en animals inoculats, infectats per contacte i infectats vacunats. Paral·lelament, es va valorar la sensibilitat en la detecció de l’excreció del virus en els FO per PCR quantitativa, prenent com a referència la virèmia. Els animals inoculats i infectats per contacte van mostrar un patró d’excreció en FO semblant, en canvi el patró dels animals vacunats va ser més inconstant. En els animals vacunats també es va produir una reducció significativa del període d’excreció (P<0.05). Els valors Kappa entre els FO i el sèrum van ser >0.68 en tots els casos. L’objectiu del segon estudi era identificar i valorar les pèrdues d’eficiència que es podrien donar durant el procés d’anàlisi de les mostres de FO per qRT-PCR: efecte de les diferents matrius per la recollida de les mostres, el temps i la temperatura fins a la seva anàlisi, les condicions de conservació, l’efecte de la centrifugació, de diferents mètodes d’extracció de l’RNA i dels reactius per la PCR. Es van utilitzar FO lliures de PRRSV infectats amb concentracions conegudes del virus. No es van trobar diferències entre les matrius de recol·lecció ni els mètodes de conservació dels FO. En canvi, es va observar una menor sensibilitat analítica (P<0.05) en les mostres mantingudes durant 72h a 4ᵒC. En les dilucions límit, la centrifugació a 15.000 g va produir un augment significatiu en la sensibilitat de les mostres (P<0.05). Els mètodes d’extracció amb majors eficiències van ser el TRIzol®, el MagMAX™ i el NuceoSpin®. En dilucions de FO iguals, el kit de qRT-PCR LSI VetMAX® va donar menors Cts que l’AgPath™. En el tercer estudi, a partir de la bibliografia existent i les dades obtingudes en els experiments anteriors, es va elaborar un càlcul de probabilitats per determinar els límits de la detecció del PRRSV1 en FO quan s’usa en granges amb baixes prevalences. Es van comparar diversos tipus de granges d’acord amb l’estructura de les maternitats, de les transicions i la mida del mostreig. Segons els resultats obtinguts, els factors determinants de la detecció del PRRSV van ser la incidència dels esdeveniments verticals, el nombre de truges en producció, la mida dels corrals de transició, així com el nombre de corrals analitzats. La qRT-PCR va ser capaç de detectar el virus en els FO en la majoria d’escenaris possibles, fins i tot quan es tenia un sol animal infectat en corrals de 50 o menys animals. En la majoria de casos, incidències de parts amb animals virèmics inferiors al 5% en granges de 1.000 o menys truges, feien que la presència d’animals infectats en els corrals de transició fos un fenomen rar, i la seva detecció depengués més del nombre de corrals mostrejats que de qualsevol altra circumstància. Com a conclusió, els resultats de la present tesi estableixen un protocol que maximitza la sensibilitat en la detecció del PRRSV1 en FO i en demostren la seva validesa en transicions amb baixes prevalences.
In recent years, the use of oral fluids (OF) for the diagnosis and monitoring of Porcine respiratory and reproductive syndrome virus (PRRSV) has increased popularity in North America while in Europe its use is still scarce. Few studies have been conducted on the performance of diagnostic techniques for PRRSV1 using OF. The general objective goal of the present thesis was to evaluate the efficiency of the use of OF in the diagnosis of PRRSV1, determining the factors influencing their diagnostic performance. The first study is aimed at evaluating at an individual level, shedding of PRRSV1 in OF over time of pigs inoculated experimentally and in naïve and vaccinated animals infected by contact with inoculated pigs. In addition, the agreement between virus shed in OF and viremia was also assessed. Inoculated and naïve pigs showed a similar pattern of shedding of PRRSV in OF while vaccinated showed a more variable and irregular pattern. The average period of shedding in OF was significantly shorter (P<0.05) in vaccinated animals compared to the others. Kappa values for the comparison of blood and OF were >0.68 in all cases. The purpose of the second study was to identify and assess the losses of efficiency during the process of analysis of OF samples by qRT-PCR: the effect of different matrices for collecting the samples, different times and temperatures until the analysis, conditions of storage, the effect of centrifugation and different PCR reagents and RNA extraction methods. OF collected from weaners of PRRSV-free commercial farms were spiked with known concentrations of PRRSV1. There were no differences between the results obtained with the different collection matrices or conservation methods. Regarding storage, a significant loss of analytic sensitivity (P<0.05) was observed when OF were stored for 72h at 4ᵒC. Centrifugation at 15,000 g increased significantly (P<0.05) the sensitivity for samples containing 100 TCID50/mL. The RNA extraction methods that resulted in the best efficiencies were TRIzol®, MagMAX™ and NucleoSpin®. At similar dilutions, the LSI VetMAX® produced less Ct for the detection of the virus in OF than AgPath™. In the third study, based on the existing literature and data obtained in previous experiments, we calculated the probability of detecting PRRSV1 when pen-based OF were used for monitoring by qRT-PCR in low prevalence farms. Different types of farms were compared according their farrowing and nursery structure, and the size of sampling. Regarding the results obtained, the most decisive factors influencing the detection of PRRSV were the incidence of vertical events, the size of the reproductive herd and the pens for weaners; and the number of pens examined. The qRT-PCR would be able to detect PRRSV in OF in most scenarios even when there was only one infected animal out of 50 penmates. The main factors which limited the detection of PRRSV were the proportion of viremic farrows and the distribution of infected animals from the farrowing units to the nurseries. In most cases, in farms with ≤1,000 sows and less than 5% of PRRSV1 incidence, the presence of infected animals in nursery pens were a rare event and the most important factor was the number of pens sampled. In conclusion, the results of the present thesis establish a protocol for optimising the sensitivity of PRRSV1 detection in OF samples. Taking into account these recommendations, OF are a good diagnostic tool for the detection of virus circulation in nurseries because PRRSV is detected even in the worst scenarios.

Tesis para optar al Grado de Magíster en Ciencias Animales y Veterinarias mención Medicina Preventiva Animal
En el año 2000 se declaró de interés nacional la erradicación de Peste Porcina Clásica (PPC) en el territorio Colombiano, por medio de la estrategia de zonificación el país estuvo libre de la enfermedad manteniendo algunas zonas con vacunación en el periodo de 2007 a 2013; sin embargo, como consecuencia del transporte ilegal de cerdos desde la frontera con Venezuela, la enfermedad reingresó avanzando a algunas zonas declaradas libres de la enfermedad. Este documento busca evaluar el impacto económico para el Estado de la reintroducción de la PPC en Colombia, simulando y cuantificando la presentación de un brote en una zona libre de la enfermedad (departamento de Antioquia) y un brote en la zona de control (departamento de Arauca) en un sistema de producción de traspatio y tecnificado siguiendo el manejo establecido por el plan de contingencia del Instituto Colombiano Agropecuario ICA y comparándolo con los costos asociados al programa de vigilancia epidemiológica para PPC en el año 2015. Producto de la simulación y valoración de escenarios, los costos de erradicación de un brote de PPC en el departamento de Arauca son menores que los costos asociados al programa de vigilancia para PPC en el año 2015 a diferencia de un brote de PPC en el departamento de Antioquia, donde los costos duplican el presupuesto del programa de vigilancia. Por lo cual, es necesario fortalecer las medidas de vigilancia y el programa de control y erradicación, con el fin de evitar pérdidas económicas para el Estado y fortalecer el estatus sanitario del país.
In 2000 it was declared as national interest the eradication of classical swine fever (CSF) in the Colombian territory, through zoning strategy the country was free from the disease by keeping some areas with vaccination between the period 2007 to 2013, but as a result of illegal transport of pigs from the border with Venezuela, the disease reentered forward to some areas declared free of the disease. This paper aims to assess the economic impact for the state of the reintroduction of the CFP in Colombia, simulating and quantifying the presentation of one outbreak in a free area of the disease (department of Antioquia) and one outbreak in the control area (Arauca) in a backyard production system and modernized following the management established by the contingency plan of the Instituto Colombiano Agropecuario ICA, and compared it to the costs associated with CSF monitoring epidemiologic program in 2015. Product of simulation and assessment of scenarios, the costs of an outbreak of CSF in the department of Arauca are less than the costs associated with the monitoring program for CSF in 2015 as opposed to an outbreak of CSF in the department Antioquia where costs double the budget of the monitoring program. Therefore it is necessary to strengthen surveillance and control program and eradication, in order to avoid economic losses for the state and strengthen the health status of the country.

L’échoendoscopie thérapeutique occupe une place croissante et incontournable de l’activité médico-chirurgicale en pathologie digestive depuis le développement des sondes sectorielles. Les domaines d’application de cette technique interventionnelle mini-invasive sont de plus en plus nombreux, d’une part en pathologie biliopancréatique et entérale permettant la réalisation d’anastomoses évitant un recours à la chirurgie traditionnelle, et d’autre part en cancérologie digestive car elle permet de délivrer un agent thérapeutique, une source de rayonnement ou des cellules liées au système immunitaire dans une lésion ciblée. L’abord vasculaire est une voie récente et prometteuse de l’échoendoscopie interventionnelle offrant des perspectives thérapeutiques en hépatologie notamment dans le domaine de l’hypertension portale. Ce travail se décompose en trois parties décrivant successivement l’état de l'art des applications de l’échoendoscopie interventionnelle, puis deux travaux originaux expérimentaux : dans un premier temps, le transfert de la technique d’anastomose biliodigestive (réalisée régulièrement dans notre centre) à l’abord vasculaire portosystémique sur animal sain, puis dans un deuxième temps la validité de la technique sur modèle animal de fibrose hépatique. La première série d’expérimentations a permis de mettre au point la technique de création d’un shunt intrahépatique portosystémique par échoendoscopie chez 23 cochons sains, en collaboration avec l’équipe de radiologie vasculaire. Ces procédures ont été réalisées dans une salle de cathétérisme vasculaire expérimentale (laboratoire Caviti) dans le cadre du laboratoire d’accueil ISIT (Image Science for Interventional Techniques, Pr. L. Boyer, Pr. J.-Y. Boire, UMR CNRS/UDA 6284). Les résultats ont été concluants puisque le shunt est apparu faisable dans 91% des cas, fonctionnel dans 81% des cas avec une morbidité de 14,2%. La deuxième série d’expérimentations a permis de valider cette technique d’échoendoscopie chez un modèle porcin de fibrose hépatique par embolisation radiologique, en collaboration avec l’Institut National de Recherche Agroalimentaire de Theix pour la stabulation des animaux. Ce travail multidisciplinaire a permis une collaboration étroite entre endoscopistes, radiologues, chirurgiens viscéraux et anatomopathologistes, s’inscrivant dans une démarche de recherche transversale. Les résultats sont encourageants puisqu’un shunt intrahépatique porto-systémique par échoendoscopie s’est révélé techniquement faisable et fonctionnel sur modèle de fibrose hépatique avec une survie à 7 jours des animaux dans deux tiers des cas. Avant d’envisager une étude princeps chez l’homme, des études complémentaires sont néanmoins nécessaires sur des modèles porcins présentant une fibrose hépatique associée à une hypertension portale en augmentant la durée de stabulation, en homogénéisant la procédure de squelettisation de l’artère hépatique et en adaptant la dose de solution injectée. Cette technique pourrait constituer à terme une alternative en cas d’échec de la technique standard pour complications de l’hypertension portale (shunt intrahépatique portosystémique par voie transjugulaire) chez des patients alors en situation d’impasse thérapeutique
Therapeutic endoscopic ultrasonography is a growing and essential part of the medical and surgical activity in digestive pathology since the development of sectoral probes. The fields of application of this minimally invasive interventional technique are more and more numerous, on the one hand in biliopancreatic and enteral pathology, allowing the realization of anastomoses without traditional surgery, and on the other hand in digestive oncology, because it makes it possible to deliver a therapeutic agent, a source of radiation or cells linked to the immune system in a targeted lesion. The vascular approach is a recent and promising way of interventional endoscopic ultrasound offering therapeutic perspectives in hepatology, especially in the field of portal hypertension.This work is divided into three parts successively describing the state of the art of the applications of interventional ultrasonoscopy, then two original experimental works: initially, the transfer of the technique of biliodigestive anastomosis (performed regularly in our center ) at the portosystemic vascular approach on healthy animal, then in a second time the validity of the animal model technique of liver fibrosis.The first series of experiments led to the development of the technique for the creation of an intrahepatic shunt portosystemic by echoendoscopy in 23 healthy pigs, in collaboration with the vascular radiology team. These procedures were performed in an experimental vascular catheterization room (Caviti laboratory) as part of the ISIT host laboratory (Image Science for Interventional Techniques, Prof. L. Boyer, Pr. J.-Y.Boire, UMR CNRS / UDA 6284). The results were conclusive since the shunt appeared feasible in 91% of cases, functional in 81% of cases with a morbidity of 14.2%. The second series of experiments validated this endoscopic ultrasound technique in a porcine model of liver fibrosis by X-ray embolization, in collaboration with Theix National Institute for Food Research for Animal Stabling. This multidisciplinary work has allowed close collaboration between endoscopists, radiologists, visceral surgeons and anatomopathologists, as part of a transversal research approach. The results are encouraging since a porto-systemic intrahepatic shunt by endoscopic ultrasonography proved to be technically feasible and functional on a hepatic fibrosis model with a 7-day survival of the animals in two-thirds of the cases. Before considering a primary study in humans, additional studies are nevertheless necessary on porcine models presenting hepatic fibrosis associated with portal hypertension by increasing the duration of stabulation, by homogenizing the procedure of skeletonization of the hepatic artery and by adjusting the dose of injected solution. This technique could eventually be an alternative in case of failure of the standard technique for complications of portal hypertension (intrahepatic portosystemic shunt transjugular) in patients then in a situation of therapeutic impasse

La polispermia (entrada de más de un espermatozoide al ovocito) es una condición patológica en mamíferos ya que impide el desarrollo embrionario. En la especie porcina, la polispermia es un problema común y aún sin resolver en los sistemas de fecundación in vitro (FIV). Los gránulos corticales (GCs) de los ovocitos de mamíferos estan implicados en el bloqueo de la polispermia, sin embargo, poco se sabe acerca de la composición y función de estas organelas. Esta ampliamente descrito que las moléculas liberadas de los GCs durante la fecundación o activación ovocitaria mediante estimulación química o eléctrica producen importantes modificaciones, particularmente en la zona pelúcida (ZP), el espacio perivitelino y, probablemente, el oolema del ovocito. Estas modificaciones repercuten directamente en el bloqueo de la polispermia. El conocimiento del contenido de los GCs y proteínas del oolema así como su papel durante la fecundación en la especie porcina es escaso. La mayoría de la información que se tiene en la actualidad se ha obtenido del modelo murino. En la presente tesis doctoral, investigamos la presencia de posibles proteínas de los GCs en ovocitos porcinos así como la presencia de metaloproteasas ADAM-10 y ADAM-17 en ovocitos porcinos y bovinos. Para ello, este estudio se dividió en tres apartados:
Polyspermy (entering of more than a spermatozoon into the oocyte) is a pathological condition in mammals since it avoids the normal embryonic development. In the pig species, Polyspermy is a common problem still unsolved in the current systems of in vitro fertilization (IVF). The cortical granules (CGs) from mammal's oocytes are involved in the block to polyspermy. However, little is known about the composition and function of these organelles. It is widely described that the molecules released of the CGs during the fertilization or oocyte activation, by means of chemical or electrical stimulation, produce important modifications, particularly in the zona pelucida (ZP), the periviteline space and, probably, oolema of the oocyte. These modifications have a direct role in the block to polyspermy. The knowledge about the content of the CGs and the oolema proteins of as well as their role during fertilization in pigs is still scarce. The majority of the information that we currently have it has been obtained from the murine model. In the present Doctoral Thesis, we investigated the presence of possible proteins of the CGs in pig oocytes as well as the presence of metaloproteases ADAM-10 and ADAM-17 in pig and bovine oocytes. For that, this study was divided into three sections

As matrizes acelulares são utilizadas na engenharia de tecidos com o objetivo da reconstrução ou substituição de tecidos danificados. Nesse trabalho matrizes acelulares foram preparadas a partir de pericárdio porcino e pericárdio bovino, por possuírem uma grande quantidade colágeno tipo I, usando-se ao tratamento de hidrólise alcalina nos tempos de 12 e 24 horas. Essas matrizes foram desenvolvidas para serem utilizadas como barreiras de proteção no processo de cicatrização de feridas como também na indução de formação de tecidos ósseos. Foram submetidas ao teste de biocompatibilidade in vivo, com a implantação das matrizes no subcutâneo de 65 ratos Wistar, machos, adultos que sofreram eutanásia aos 7, 14, 21, 28 e 35 dias de pós-operatório. A avaliação microscópica no 7º dia mostrou existência da matriz e uma grande quantidade de infiltrado inflamatório em volta do material, sendo que ao 14º dias além do infiltrado inflamatório, foi observado uma neovascularização. Aos 21º e 28º dias foi observado uma diminuição gradual do infiltrado inflamatório, crescimento celular no local em que as fendas se formaram nas matrizes e uma grande revascularização. Ao 35º dias a matriz de pericárdio bovino e porcino foram reabsorvidas totalmente. Comparando o pericárdio bovino com o porcino tratado por 12 e 24 horas em relação ao tempo de degradação, foi observado que as matrizes tanto bovina como a porcina tratadas por 24 horas degradaram mais rápido que as tratadas por 12 horas. Portanto as matrizes de pericárdio bovino e porcino são biocompativeis, pois não causaram reações citotóxicas, reações inflamatórias e nem imunológica adversa.
Acellular matrices are used in tissue engineering for reconstruction or replacement of damaged tissues .In this study acellular matrices were developed using porcine pericardium and bovine pericardium, which contain a large quantity of type I collagen, using an alkaline hydrolysis treatment at the times of 12 hours and 24 hours. The matrices were developed to be used as protective barriers in the process of wound healing, and also in bone tissue inducing formation. In the test of in vivo biocompatibility, matrices were implanted subcutaneously in 65 adult male Wistar rats, sacrificed at 7, 14, 21, 28 and 35 days postoperatively. The microscopic evaluation on day 7 showed a large amount of inflammatory infiltrate around the material; at the 14th day, beyond the inflammatory infiltrate was observed a neovascularization. At 21st and 28th days were observed a gradual decrease in the inflammatory infiltrate, cellular growth and a large neovascularization. By day 35, porcine and bovine pericardium matrices were completely resorbed. Comparing the degradation time between the matrices treated with 12 and 24 hours, it was observed that both matrices treated for 24 h, degraded faster than those treated for 12 hours. Both matrices are biocompatible, do not cause cytotoxic reactions, inflammatory or immune adverse reactions.

Memoria para optar al Título Profesional de Médico Veterinario
La presente memoria de título buscó identificar y valorizar los principales impactos económicos asociados a la presencia de Peste Porcina Clásica (PPC) en países de la subregión andina, considerando de que se trata de la enfermedad porcina más importante en términos económicos, y desde el año 2000 está bajo un plan continental de erradicación a cargo de la Organización de las naciones unidas para la alimentación (FAO). Para poder realizar la valoración de los impactos, se hizo necesario conocer el cuerpo normativo que rige los programas de control y erradicación de la PPC en países de la subregión andina, identificar los tipos de pérdidas que genera la presencia de la enfermedad, y finalmente determinar la cantidad de factores de producción que se ven afectados en presencia de la enfermedad. Se recopilaron datos mediante revisión bibliográfica y mediante el uso de documentos de trabajo interno de la FAO. Adicionalmente se realizaron encuestas a expertos de 6 países, y se realizaron entrevistas a profesionales encargados de los Servicios Veterinarios Oficiales (SVO) de los países de la subregión andina. La investigación evidenció una ausencia de estandarización de conceptos entre países, lo que dificultó los análisis comparativos, junto con una muy baja existencia de datos previos sobre pérdidas económicas y la ausencia de registro de las pérdidas generadas por la PPC en la subregión. Lo anterior obligó a destinar muchos esfuerzos a la generación de información primaria de las pérdidas a través de expertos y SVO, que finalmente permitió tener una primera aproximación hacia datos cuantitativos. Los datos obtenidos mediante la presente memoria, otorgan una base para el análisis y para la generación de simulaciones del impacto de PPC, lo que permitirá a los SVO tomar decisiones con una mayor cantidad de antecedentes, menor incertidumbre, y mayor eficiencia económica.
Proyecto FAO, TCP/RLA/3305

Tesina sobre la inseminación artificial en cerdos
El presente documento se orientará a ser una recapitulación de información sobre la colecta, dilución e inseminación artificial en la especie porcina, para tener una versión actualizada de los procesos técnicos a seguir para una efectiva inseminación porcina, tanto para estudiantes, técnicos y especialistas en el área porcícola.

El objetivo de la tesis aquí presentada fue el de proporcionar información sobre la epidemiología del circovirus porcino tipo 2 (PCV2). Los cuatro estudios incluidos en esta Tesis Doctoral se resumen a continuación: El primer estudio fue dirigido a averiguar la prevalencia de PCV2 y otros virus porcinos, en concreto el virus reproductivo y respiratorio porcino (PRRSV), el virus de la influenza porcina (SIV), el virus de la enfermedad de Aujeszky (ADV) y parvovirus porcino (PPV) en granjas porcinas españolas. Se obtuvo que a principio-mitad de los 2000, PCV2 y PPV mostraron evidencias de una distribución ubicua en cerdos; PRRSV y SIV también estaban extendidos. La seroprevalencia del virus salvaje de ADV disminuyó con el tiempo. La seroprevalencia en verracos era inferior que en madres y engorde. El Segundo trabajo consistió en un estudio exploratorio de casos-controles dirigido a encontrar factores de riesgo que, en asociación con la infección por PCV2, inducían la expresión de la enfermedad sistémica por PCV2 (ES-PCV2), una enfermedad multifactorial. Se concluyó que la infección temprana por PCV2, medida por la evidencia de seroconversión, es un factor predisponente para el desarrollo de ES-PCV2. El tercer estudio se enfocó a los antecedentes genéticos, un factor de riesgo específico para la ES-PCV2. Se concluyó que los antecedentes genéticos son un factor de riesgo para el desarrollo de ES-PCV2. Los lechones procedentes de verracos Pietrain mostraron el mejor rendimiento clínico seguido de los de verracos Large White x Pietrain. Los lechones de verracos Large White x Duroc fueron los más afectados por ES-PCV2. Finalmente, el último estudio fue dirigido a averiguar el efecto de la carga de PCV2 en suero en la ganancia media diaria de peso (ADWG) durante el periodo post-destete. Se concluyó que la variación de ADWG entre cerdos en granjas afectadas por ES-PCV2 está parcialmente explicada por la cantidad de PCV2 en suero desde el destete al sacrificio. Se identificaron 3 subpoblaciones de cerdos con diferentes cargas de PCV2 en este periodo. Estas subpoblaciones experimentaron diferentes ADWG, donde cuanto mayor era la carga de PCV2 menor era el ADWG.
The present thesis aimed to provide information on porcine circovirus type 2 (PCV2) epidemiology. The four studies included in this PhD Thesis are summarised below: The first study aimed to assess the prevalence of PCV2 and other swine viruses, namely reproductive and respiratory syndrome virus (PRRSV), swine influenza virus (SIV), Aujeszky’s disease virus (ADV) and porcine parvovirus (PPV) in Spanish pig herds. It was obtained that in the early-mid 2000s, PCV2 and PPV showed evidence of ubiquitous distribution in pigs; PRRSV and SIV were also widespread. Seroprevalence against ADV wild virus decreased over time. Boar studs had lower seroprevalences than sow and fattening herds. The second work consisted in an exploratory case-control study aimed to assess risk factors that, in association with PCV2 infection, induced the expression of porcine circovirus type 2-systemic disease (PCV2-SD), a multifactorial disease. It was concluded that early infection by PCV2, measured by evidence of seroconversion, is a predisposing factor for PCV2-SD occurrence. The third study focused on the pig genetic background, a specific risk factor for PCV2-SD. It was concluded that the genetic background is a risk factor for PCV2-SD development. Piglets from pure Pietrain boars showed the best clinical performance followed by piglets from Large White x Pietrain boars. Piglets from Large White x Duroc boars were the most affected by PCV2-SD. Finally, the last study aimed to assess the effect of PCV2 loads in pig serum on average daily weight gain (ADWG) during the postweaning period. It was concluded that ADWG variation among pigs in PCV2-SD affected farms is partly explained by serum PCV2 load from weaning to slaughter age. Three subpopulations of pigs with different serum PCV2 loads from weaning to slaughter age were identified. These subpopulations experienced significantly different ADWG, in which the higher the PCV2 load the lower the ADWG.

La regulació de l’engreixament porcí i la qualitat de la carn són encara poc coneguts. Inicialment, vam investigar la variabilitat de gens candidats localitzats a regions QTL associades amb caràcters de qualitat de la carn i contingut i composició de greix intramuscular. Vam identificar polimorfismes en aquests gens candidats a partir de dades d’RNA-seq i seqüències de genoma complet (WGS) de cinc porcs Duroc. El genotip d’ATP1A2 va ser significativament associat a la conductivitat elèctrica (CE) del múscul longissimus dorsi (LD) a nivell cromosòmic. Els nostres resultats suggereixen que el gen ATP1A2 pot estar involucrat en la regulació de la CE en el múscul. D’altra banda, vam fer ús de les dades de WGS per localitzar mutacions stop gained segregant en una població Duroc (Lipgen). Van ser identificats set porcs homozigots per una mutació potencialment letal en el gen ASS1. Després de seqüenciar aquesta regió a nivell genòmic i transcriptòmic, es va revelar la presència d’una mutació immediatament abans, que eliminaria el codó d’aturada, generant un polimorfisme dinucleotídic que causa un canvi benigne d’aminoàcid en la seqüència d’ASS1. Seguidament, vam utilitzar dades prèvies d’expressió diferencial d’RNA-seq per investigar l’associació de gens candidats amb caràcters de qualitat de la carn. Dos polimorfismes localitzats en els gens CRY2 i MIGA2 van mostrar associacions significatives amb el contingut d’àcid esteàric en LD, i amb la concretació d’LDL en sèrum, respectivament. Aquests polimorfismes també van ser associats amb l’expressió dels seus gens respectius. Anàlisis a nivell cromosòmic van mostrar que aquests polimorfismes poden no ser els SNPs casuals. També vam analitzar els polimorfismes localitzats en gens microRNA a partir d’un total de 120 WGS de porcs domèstics i salvatges d’Àsia i Europa. La variabilitat de regions miRNA va ser molt reduïda en la seed comparat amb altres regions del miRNA i amb la resta del genoma. Quinze SNPs en gens miRNA van ser genotipats en la població Lipgen. Els resultats van revelar, entre d’altres, una variant localitzada en el bucle apical d’ssc-miR-326 significativament associada amb l’expressió d’alguns dels seus mRNAs diana. Aquest SNP pot contribuir a la reestructuració de l’aparellament de bases a la forquilla del miRNA, modificant l’eficiència de la maduració del propi miRNA. Altrament, ens vam proposar millorar l’encara limitada anotació del miRNAoma porcí mitjançant el desenvolupament d’un pipeline bioinformàtic per a la identificació i anotació de gens miRNA. La fracció d’RNA petit de 48 porques Duroc va ser seqüenciada per detectar miRNAs nous i ja coneguts. Els transcrits de dades d’small RNA-seq i miRNAs madurs anotats en humà van ser cartografiades en el genoma porcí. Es van reconstruir seqüències candidates mitjançant la cerca de motius nucleotídics. Es van obtenir un conjunt de paràmetres de seqüència i termodinàmics de cada seqüència i es va fer servir un algoritme de Machine Learning basat en grafs per predir miRNAs, tant nous com coneguts. Van ser descoberts un total de 47 miRNAs porcins putatius. L’expressió de tres d’ells va ser avaluada mitjançant tècniques d’RT-qPCR i confirmada en una població independent de porcs de raça Göttingen minipig. Finalment es van utilitzar les dades d’small RNA-seq per determinar miRNAs diferencialment expressats (DE) entre porques en dejú i després de rebre aliment. Les xarxes de regulació gènica d’interaccions miRNA-mRNA van revelar mòduls de coexpressió de gens relacionats amb el metabolisme de lípids. A més, es va evidenciar la potencial influència de miRNAs DE en regular l’expressió de mRNAs amb funcions en el metabolisme de la glucosa i l’homeòstasi energètica.
La regulación del engrasamiento porcino y la calidad de la carne son aún poco conocidos. Inicialmente, investigamos la variabilidad de genes candidatos localizados en regiones QTL asociadas con caracteres de calidad de la carne y contenido y composición de grasa intramuscular. Identificamos polimorfismos en dichos genes candidatos a partir de datos de RNA-seq y secuencias de genoma completo (WGS) de cinco cerdos Duroc. El genotipo de ATP1A2 fue significativamente asociado a la conductividad eléctrica (CE) del músculo longissimus dorsi (LD) a nivel cromosómico. Nuestros resultados sugieren que el gen ATP1A2 puede estar involucrado en la regulación de la CE en el músculo. Por otra parte, hicimos uso de los datos de WGS para identificar mutaciones stop gained segregando en una población Duroc (Lipgen). Siete cerdos homocigotos para una mutación potencialmente letal en el gen ASS1 fueron identificados. Tras secuenciar dicha región a nivel genómico y transcriptómico, se reveló la presencia de una mutación inmediatamente antes, que eliminaría el codón de parada, generando un polimorfismo dinucleotídico que causa un cambio benigno de aminoácido en la secuencia de ASS1. Seguidamente, utilizamos datos previos de expresión diferencial de RNA-seq para investigar la asociación de genes candidatos con caracteres de calidad de la carne. Dos polimorfismos localizados en los genes CRY2 y MIGA2 mostraron asociaciones significativas con el contenido de ácido esteárico en LD, y con la concentración de LDL en suero, respectivamente. Estos polimorfismos también fueron asociados con la expresión de sus respectivos genes. Análisis a nivel cromosómico mostraron que estos polimorfismos pueden no ser los SNPs causales. Además, analizamos los polimorfismos localizados en genes microRNA a partir de un total de 120 WGS de cerdos domésticos y salvajes de Asia y Europa. La variabilidad de regiones miRNA estuvo muy reducida en la seed comparado con otras regiones del miRNA y con el resto del genoma. Quince SNPs en genes miRNA fueron genotipados en la población Lipgen. Nuestros resultados revelaron, entre otros, una variante localizada en el bucle apical de ssc-miR-326 significativamente asociada con la expresión de algunos de sus mRNAs diana. Este SNP puede contribuir a la reestructuración del apareamiento de bases en la horquilla del miRNA, modificando la eficiencia de la maduración del propio miRNA. Además, nos propusimos mejorar la aún limitada anotación del miRNAoma porcino mediante el desarrollo de un pipeline bioinformático para la identificación y anotación de genes miRNA. La fracción de RNA pequeño de 48 cerdas Duroc fue secuenciada para detectar miRNAs nuevos y ya conocidos. Los transcritos de datos de small RNA-seq y miRNAs maduros anotados en humano fueron cartografiados en el genoma porcino. Se realizó la reconstrucción de secuencias candidatas mediante la búsqueda de motivos nucleotídicos. Se obtuvieron un conjunto de parámetros de secuencia y termodinámicos de cada secuencia y se empleó un algoritmo de Machine Learning basado en grafos para predecir miRNAs, tanto nuevos como conocidos. Un total de 47 miRNAs porcinos putativos fueron detectados. La expresión de tres de ellos fue evaluada mediante técnicas de RT-qPCR y confirmada en una población independiente de cerdos de raza Göttingen minipig. Finalmente se utilizaron los datos de small RNA-seq para determinar miRNAs diferencialmente expresados (DE) entre cerdas en ayunas y tras recibir alimento. Las redes de regulación génica de interacciones miRNA-mRNA relevaron módulos de co-expresión de genes relacionados con el metabolismo de lípidos. Además, se evidenció la potencial influencia de miRNAs DE en regular la expresión de mRNAs con funciones en el metabolismo de la glucosa y la homeostasis energética.
The genetic modulators of porcine fatness and meat quality traits, as well as their mechanisms of action, are still poorly understood. First, we investigated the variability of candidate genes located within QTL regions associated with meat quality traits and intramuscular fat content and composition. Polymorphic sites located at candidate genes were identified based on RNA-seq data and whole-genome sequencing of five Duroc boars. Significant association between ATP1A2 genotype and electric conductivity (CE) in the longissimus dorsi (LD) muscle, as well as in a chromosome-wide analysis, were revealed. Our results suggest that the ATP1A2 gene might be involved in the regulation of the CE of the skeletal muscle. Moreover, we employed whole-genome sequencing data from the five Duroc boars to identify putative stop gained mutations segregating in a Duroc population (Lipgen). Seven pigs homozygous for a potentially lethal nonsense recessive mutation in the ASS1 gene were detected. After sequencing such region at the genomic and transcriptomic levels, the presence of an additional polymorphism located immediately before the nonsense mutation that disrupts the stop codon was revealed, forming a dinucleotide polymorphism that causes a benign amino acid substitution in the ASS1 sequence. Furthermore, we used previous RNA-seq differential expression data to investigate the association of candidate genes with meat quality traits. Two polymorphisms located in the CRY2 and MIGA2 genes showed significant associations with stearic acid content in LD and with LDL serum concentration, respectively. These SNPs were also associated with the mRNA levels of the corresponding genes. Joint chromosome-wide association analyses showed that these polymorphisms are not the ones showing the most significant associations. We also studied polymorphisms residing in microRNA genes. A total of 120 whole-genome sequences from European and Asian wild boars and domestic pigs were used for variant calling analyses, and polymorphisms within miRNA loci were investigated. Variability within miRNA loci was strongly reduced in the seed region compared with the rest of the miRNA sequence and other regions in the genome. Fifteen SNPs mapping to miRNA genes were genotyped in the Lipgen population. Our results revealed, among others, one variant located in the apical loop of ssc-miR-326 as significantly associated with the expression of some of its targets. This SNP might contribute to a structural rearrangement of the miRNA hairpin pairing, thus modifying the efficiency of the miRNA maturation. Subsequently, we aimed to improve the yet poorly annotated porcine miRNAome by developing a bioinformatic pipeline for the discovery and annotation of miRNA genes. The small RNA fraction of 48 Duroc gilts was sequenced and used to detect novel and known expressed miRNAs. Small RNA-seq transcripts and annotated human mature miRNAs were mapped to the porcine genome. Reconstruction of candidate hairpin sequences was performed by applying a motif search correction approach. A series of sequence and thermodynamic features were obtained from each sequence and a Machine-Learning graph-based transductive algorithm was employed for predicting novel and annotated miRNA sequences. A total of 47 unreported putative porcine miRNAs were detected with this approach. The expression of three of the unreported miRNAs was assessed by using RT-qPCR analyses and their expression in an independent Göttingen minipig population was confirmed. Finally, we employed the muscle small RNA-seq data set to determine differentially expressed (DE) miRNAs between fasting and fed pigs. Gene regulatory networks for miRNA-mRNA interactions highlighted co-expression modules containing lipid-related genes. The potential influence of several DE miRNAs in regulating the expression of mRNA genes with key roles in glucose metabolism and energy homeostasis was evidenced.

El descobriment dels microRNAs (miRNAS) com a nous reguladors de l’expressió gènica ha obert un nou camp en l’estudi de quin és el comportament d’aquests RNAs de mida petita i descriure en quins processos actuen i de quina manera regulen l’expressió gènica. L’aparició de les tècniques de seqüenciació massiva ha permès la descripció i estudi dels perfils d’expressió de miRNAs en diferents situacions, a fi d’observar la seva implicació en els processos biològics, tant fisiològics com patològics. Aquesta tesi exemplifica, mitjançant dues aproximacions diferents, l’ús d’aquestes tècniques per a la descripció, descobriment i estudi de perfils de miRNAs en l’espècie porcina. La primera part del treball es va dissenyar amb l’objectiu d’ampliar el nombre de miRNAs descrits en el porc. Les aproximacions utilitzades per a la determinació de nous miRNAs varen ser, primer de tot, la descripció del perfil d’expressió de miRNAs en el ronyó del porc, incloent els miRNAs ortòlegs i, segon, l’ús d’un protocol de descobriment i validació de nous miRNAs porcins. Una altra motivació del treball va ser el fet d’estudiar possibles canvis en l’expressió dels miRNAs en races de porc de diferents orígens, des de races europees fins a asiàtiques, incloent races europees amb influència asiàtica, on hem descrit miRNAs diferencialment expressats i s’ha estudiat la seva possible funcionalitat. En la segona part del treball es va dur a terme una infecció experimental amb el virus de la malaltia d’Aujeszky (ADV), l’herpesvirus porcí tipus 1 (SHV-1), amb una soca virulenta (NIA-3) i una soca vacunal (Begonia). Es varen realitzar dues aproximacions diferents: un aproximació in vitro utilitzant les línies cel·lulars PK-15 derivades del ronyó de porc, i una aproximació d’infecció experimental in vivo utilitzant el bulb olfactori i el gangli trigemin com a teixits diana per l’estudi. Amb l’objectiu d’estudiar la implicació dels miRNAs, tant virals com de l’hoste, en les interaccions hoste – patogen durant una infecció vírica, s’han descrit els perfils de miRNAs i s’han avaluat les diferències en la seva expressió, no només entre el grup infectat i el grup control, sinó també entre les soques utilitzades i entre les dues aproximacions. També s’han descrit nous miRNAs virals que s’expressen durant la infecció. Finalment, s’ha elaborat, amb estudis funcionals in silico, una xarxa d’interaccions entre els miRNAs virals, els miRNAs de l’hoste diferencialment expressats i els gens que codifica l’agent infecciós SHV-1. Donada la importància de la tècnica de l’RT-qPCR per a la validació de l’expressió de miRNAs, també s’ha realitzat un estudi addicional per avaluar l’estabilitat en l’expressió d’alguns miRNAs per a poder ser utilitzats com a gens de referència en estudis de quantificació relativa de dades d’RT-qPCR, ja que fins l’actualitat s’han fet servir poc per a aquest propòsit.
The discovery of microRNAs (miRNAs) as novel gene expression regulators has opened a new field in the study of the roles of these small RNAs, as well as on describing in what processes they act and how they regulate gene expression. The emergence of next generation sequencing methods has allowed the description and study of miRNA expression profiles in different situations, in order to observe its involvement in biological processes, both physiological and pathological. This thesis illustrates, through two different approaches, the use of these techniques for the description, discovery and study of miRNA profiles in the porcine species. The first part of the study was designed with the aim of increasing the number of described miRNAs in pigs. The approaches used for the determination of novel miRNAs were, first of all, the expression profiling of miRNAs in the swine kidney, including the orthologous ones and, second, using a pipeline for the discovery and validation of new porcine miRNAs. Another motivation of this work was to study the possible changes in the kidney miRNAs expression patterns among pig breeds from different origins, from European to Asian breeds, including European breeds with Asian influences. In this sense, differentially expressed miRNAs have been described and their functional roles have been studied. In the second part of the study, an experimental infection with Aujeszky's disease virus (ADV), also known as suid herpesvirus type 1 (SHV-1), was carried out, using a virulent strain (NIA-3) and a vaccine strain (Begonia). Two different approaches were conducted: an in vitro approach using PK-15 cell lines, derived from pig kidney, and an in vivo approach using the olfactory bulb and trigeminal ganglia as target tissues. With the aim of studying the role of both host and viral miRNAs in host – pathogen interactions during SHV-1 infection, miRNAs expression profiles have been described and their expression differences evaluated, not only between infected and mock-infected groups, but also between strains and between the two performed approaches. New viral miRNAs have been described and their expression during the infection has been confirmed. Finally, a network of interactions between viral miRNAs, host differentially expressed miRNAs and SHV-1 encoded genes was developed using in silico functional studies. Given the importance of RT-qPCR method for validating the expression of miRNAs, we also performed an additional study to assess the expression stability of some miRNAs to be used as reference genes in relative quantification studies of RT-qPCR data, which they have not been widely used for this purpose.

Memoria para optar al Título Profesional de Médico Veterinario
La irrupción de la Encefalopatía Espongiforme Bovina en varios países europeos provocó un aumento en las preocupaciones respecto al tema de la seguridad alimentaria. Como consecuencia, se establecieron procedimientos de trazabilidad alimentaria, basados en una red de información capaz de afrontar los diferentes problemas acaecidos. Lo anterior, sumado a las nuevas exigencias del mercado exportador, trajo como consecuencia en la industria porcina nacional la necesidad de disponer de normas específicas sobre el tema, lo que se tradujo en la formulación de un Manual de Trazabilidad Porcina, patrocinado por la Asociación Gremial de Productores de Cerdo de Chile (ASPROCER). El presente trabajo comparó el modelo propuesto por ASPROCER, respecto a otros modelos internacionales, determinando similitudes y diferencias. Complementariamente, se realizó una encuesta a profesionales especialistas en el rubro, quienes participan y colaboran con dicha agrupación gremial, a modo de determinar su nivel de aproximación, aplicación y conocimiento respecto de dicho Manual. Entre sus resultados fundamentales destacan: a) Respecto al nivel de identificación de cerdos, existe una tendencia generalizada a realizar este proceso de manera individual; b) Dentro del estudio se presentaron serias diferencias de modelos respecto a los procesos de trazado propuestos; c) Frente el tiempo de respuesta, a las auditorías, al recall y a las relaciones humanas, se probó la existencia de poca información en comparación con el Manual preparado por ASPROCER y que éste, a su vez, es muy amplio para definir una única postura; d) A nivel país, no existe una cohesión entre el ámbito público-privado que apoye o valide el Manual de Trazabilidad Porcina; y, e) Se determinó por medio de la encuesta que aún no existe un adecuado nivel de conocimiento, aproximación y aplicación de este Manual a nivel de los encuestados. Por tanto, queda en evidencia que es necesario una mayor difusión de dicho manual, tanto a nivel de asociados como colaboradores, dentro de ASPROCER

This research examines how, when and why the Romans assigned and defined the tasks of preparing and commanding fleets during the Republic. In doing so, it brings new evidence to bear on the wider debates about the nature of the prouincia and the institutional and administrative development of the Roman empire. The communis opinio is that a prouincia originally represented a functional “sphere of operation” that was allotted or assigned to a magistrate and that it only later developed a geographical meaning with territorial connotations through the process of “provincialisation.” This research challenges that view through an analysis of the evidence for the definition, assignment and practical use of the prouincia classis and other prouinciae connected with the command of fleets. Drawing upon and analysing the lists of administrative arrangements to be found in the “annalistic” sections of the surviving books of Livy’s History, it argues that prouinciae were defined in specific geographical and functional terms long before the development of permanent territorial empire. This offers a new perspective which points to and elucidates the flexible use of the prouincia as a means of separating magistrates and promagistrates in space or by function in space. It argues that the rationale for this was to limit conflicts between commanders over command and triumphal rights. By combining evidence from a wide range of sources after the loss of Livy’s History from 167, the research shows that the above rationale for demarcating prouinciae still applied in the first century B.C. However, it also demonstrates that there were significant changes with the assignment of vast Mediterranean-wide naval prouinciae in the first half of the first century B.C. It argues that the definition of these prouinciae was made possible by the development of a singular collective Mediterranean-wide ora maritima, which was brought about by the Romans’ increasing “acknowledgement of empire.” The negative political and institutional implications of these developments are also assessed. Finally, in discussing the above, this research also provides new insights into the role and auctoritas of the Senate, the function and freedom of magistrates, and the Romans’ conceptualisation of their empire.

As lesões na pele, quando extensas, representam um grande problema de saúde pública e elevados custos associados aos tratamentos. A laserterapia tem sido aplicada por décadas a fim de acelerar o processo de cicatrização cutânea. Já a matriz de pericárdio porcino apresenta potencial para ser utilizado como curativo biológico, porém o patch tem rápida biodegradação, por isso adicionou-se quitosana. Muitos autores observaram esses tratamentos individualmente, no entanto não há dados concisos sobre a associação dessas terapias. O objetivo desse estudo foi avaliar a resposta clínica, histológica e histomorfométrica da ação fotobiomoduladora do laser no processo de reparação de úlceras cutâneas com o auxílio do curativo de pericárdio porcino revestido com quitosana. A úlcera cutânea foi induzida cirurgicamente em 48 ratos distribuídos em 6 grupos distintos com 8 animais cada, sendo eles: Matriz (curativo de pericárdio porcino); ML (curativo de pericárdio porcino com a aplicação da laserterapia); MQ (curativo de pericárdio porcino revestido com quitosana); MQL (curativo de pericárdio porcino revestido com quitosana com a aplicação da laserterapia); Ctrl (o grupo controle que não recebeu curativo algum) e Laser (o grupo controle que recebeu somente a laserterapia). Nos grupos que receberam imediatamente após o procedimento cirúrgico a radiação laser, com as seguintes características: comprimentos de onda da luz = 660nm, irradiância = 30mW/\'CM POT.2\', e densidade de energia de 30j/\'CM POT.2\'. O registro fotográfico das úlceras foi realizado no 10º dia pós-operatório e para fornecer a área ulcerada foi utilizado o software Image J. Foi realizada uma análise histológica e histomorfométrica (contagem de fibroblastos, células inflamatórias, vasos sanguíneos e colágenos) a partir de biópsias coletadas no 10º dia pós-operatório. Os resultados obtidos foram dados através das médias dos grupos com relação a cada análise, sendo eles: área da lesão (Laser = 0,48 \'CM POT.2\', Matriz = 0,46 \'CM POT.2\', Ctrl = 0,28 \'CM POT.2\', ML = 0,23 \'CM POT.2\', MQ = 0,22 \'CM POT.2\' e MQL = 0,19 \'CM POT.2\'); formação colagênica (Laser = 54,51%, Matriz = 51,75%, Ctrl = 58,84%, ML = 68,77%, MQ = 69,50% e MQL = 71,72%); células inflamatórias (Laser = 126,31, Matriz = 132,40, Ctrl = 101,85, ML = 97,67, MQ = 95,27 e MQL = 94,15); vasos sanguíneos (Laser = 6,83, Matriz = 4,48, Ctrl = 5,37, ML = 7,46, MQ = 5,42 e MQL = 3,98) e fibroblastos (Laser = 55,83, Matriz = 71,31, Ctrl = 78,63, ML = 85,40, MQ = 105,06 e MQL = 108,58). Os dados foram analisados pela análise de variância, com post hoc de Tukey, a fim de comparar os grupos com um nível de significância de 95% (p<0,05). Com base nesses achados, observou-se que houve melhora estatisticamente significante no grupo com curativos de pericárdio porcino revestido com quitosana associado à aplicação da laserterapia (MQL) quando comparado ao controle, mostrando-se mais eficaz no processo cicatricial de úlceras.
Large skin injuries represent a major public health issue, with high costs associated with the treatments. Laser therapy has been used for decades to accelerate the process of skin healing. The porcine pericardium matrix has the potential to be used as a biological dressing, but the patch has rapid biodegradation, so chitosan was added to delay this effect. Many authors have used these treatments individually; however there is no accurate data on the combination of these therapies. The aim of this study was to assess the clinical, histological and histomorphometric photobiomodulation laser action in the repair of skin ulcers associated with porcine pericardium dressing coated with chitosan. The skin ulcer was surgically induced in 48 rats, which were distributed into 6 groups with 8 animals each: Matrix (porcine pericardium dressing), ML (porcine pericardium dressing with laser therapy), MQ (porcine pericardium dressing coated with chitosan), MQL (porcine pericardium with a bandage coated with chitosan and laser therapy), Ctrl (the control group did not receive treatment) and Laser (the control group that received only laser therapy). The laser radiation groups received the treatment immediately after the surgery, with the following characteristics: light wavelength = 660nm, irradiance = 30mW/\'CM POT.2\' and of energy density 30j/\'CM POT.2\'. The photographic record of the ulcers was performed on the 10th postoperative day and to provide the ulcerated area was used the software ImageJ. We performed a histological and histomorphometric (count of fibroblasts, inflammatory cells, blood vessels and collagen) from biopsies also on the 10th postoperatively day. The results were given as means of groups from each analysis, as follows: lesion area (Laser = 0,48 \'CM POT.2\', Matriz = 0,46 \'CM POT.2\', Ctrl = 0,28 \'CM POT.2\', ML = 0,23 \'CM POT.2\', MQ = 0,22 \'CM POT.2\' e MQL = 0,19 \'CM POT.2\'), collagen formation (Laser = 54,51%, Matriz = 51,75%, Ctrl = 58,84%, ML = 68,77%, MQ = 69,50% e MQL = 71,72%), inflammatory cells (Laser = 126,31, Matriz = 132,40, Ctrl = 101,85, ML = 97,67, MQ = 95,27 e MQL = 94,15); blood vessels (Laser = 6,83, Matriz = 4,48, Ctrl = 5,37, ML = 7,46, MQ = 5,42 e MQL = 3,98) and fibroblasts (Laser = 55,83, Matriz = 71,31, Ctrl = 78,63, ML = 85,40, MQ = 105,06 e MQL = 108,58). Data were analyzed by analysis of variance with Tukey\'s post hoc to compare the groups with a significance level of 95% (p<0.05). Based on these findings, we observed that there was a statistically significant improvement in the group with porcine pericardium dressings coated with chitosan associated with the application of laser therapy (MQL) compared to control, displaying more effective in healing ulcers.

El presente manual se elabora con el propósito de brindar una guía al personal que labora en la subsección porcina de la granja SEDENA No. 1, “La Estancia, Querétaro”, buscando establecer un orden en las actividades de manejo zootécnico para promover el bienestar, la salud animal y eficientar la producción mediante la aplicación de buenas prácticas de manejo. La granja SEDENA, es una instalación dedicada a la producción de cerdos para abasto mediante un sistema de ciclo completo, además, de manejar otras especies como bovinos productores de leche, aves de postura y ciervo rojo. El trabajo se llevó a cabo mediante la búsqueda, selección y recuperación de información especializada para la integración del documento final que incluye los siguientes capítulos: Análisis situacional de la granja y Manual de operación; ambos con subtítulos que abordan diferentes aspectos como genética y reproducción, nutrición, infraestructura y medidas zoosanitarias. Los parámetros productivos de la granja SEDENA se encuentran dentro de los estándares manejados en la mayoría de las explotaciones porcinas, sin embargo, debido a un manejo zootécnico inadecuado, los resultados productivos no arrojan las ganancias económicas para el mantenimiento redituable de la explotación, condición que se maximiza debido a que no existen lineamientos que rijan las actividades de manejo en las diferentes áreas, omitiéndose el establecimiento y la aplicación de buenas prácticas pecuarias.

Currently, the in vitro production (IVP) of porcine embryos suffers from low efficiency and reduced blastocyst quality. Poor outcomes from in vitro matured oocytes and in vitro fertilised embryos have limited the use of assisted reproductive technologies (ARTs) within commercial porcine herds, reducing the potential for global genetic improvement programs. It is believed that this reduced developmental competency compared to in vivo embryos is attributable to altered metabolism resulting from in vitro culture. Improper or incomplete metabolic support from the culture media leads to production of inferior embryos. Much of the prior research centres on metabolism of carbohydrates by oocytes and embryos, with the formulation of media based on this knowledge. However, oocytes and embryos also contain endogenous lipid substrates, and there is a lack of understanding as to how and when these stores are utilised. Lipids are a dense form of energy storage, and there is evidence of their metabolism by oocytes and embryos for energy generation. Porcine oocytes and embryos have higher intracellular lipid content than other domestic livestock species, and this makes them an excellent model for studying aspects of lipid metabolism in vitro. The aim of this study was to examine the impact of lipid metabolism on the acquisition of developmental competence during porcine IVP, and how this is affected by the presence or absence of exogenous carbohydrates. Stimulation or inhibition of the β-oxidation pathway was used to discern the importance of fatty acid oxidation to oocyte maturation and embryo development during in vitro maturation (IVM), in vitro fertilisation (IVF) and in vitro embryo culture (IVC). During IVM, it was identified that porcine oocytes are capable of using different substrates to compensate for deficiencies in others. While pyruvate and glucose are preferentially utilised to support maturation, upregulation of β-oxidation can compensate for a low glucose concentration and an absence of pyruvate to support nuclear maturation. Although there was no discernible decrease in lipid content associated with this, lipids provide such a dense energy reserve that any usage may have been beyond the limit of detection. Inhibition of β-oxidation in the absence of carbohydrates had a greater effect on nuclear maturation compared to inhibition in complete media. This indicates that lipid metabolism plays a minor role during oocyte maturation in the presence of carbohydrates and is likely to be more important when other substrates are deficient. Energy generation prior to fertilisation is an important factor in the developmental outcomes of subsequent embryos. Upregulation of β-oxidation for the duration of IVF increased cleavage rates, but doses above 6mM L-carnitine led to decreased blastocyst development. This effect may be attributable to the antioxidant activity of L-carnitine, with low levels of reactive oxygen species (ROS) being required at fertilisation for normal sperm function and sperm-oocyte interactions. Oocyte incubation in media supplemented with 3mM L-carnitine for an hour prior to insemination increased cleavage and improved cryosurvival of Day 7 embryos after vitrification. While ATP content of oocytes did not increase over this period, it is unclear if lipid content was reduced. Previous studies have shown that L-carnitine treatment of oocytes and embryos decreased lipid content, thereby increasing cryotolerance. It would therefore appear that there is a limited role for β-oxidation during the IVF period itself, although upregulation immediately prior to fertilisation may have beneficial effects on metabolic processes and may provide antioxidant protection leading to improved development in early cleavage stage embryos. During embryo culture, there was a greater effect of upregulating lipid metabolism seen in the absence of carbohydrate substrates than in complete media. However, this could not support embryo development to the same extent as carbohydrate substrates. Changing nutrient requirements of embryos has led to the development of sequential media, leading to the production of better quality IVP embryos. Upregulation of β-oxidation for the first three days of culture in a single media system increased embryo quality to the same extent as a sequential carbohydrate media system, implying there is some level of plasticity to embryo metabolism allowing for adaptability to different substrates. Inclusion of L-carnitine for either a three day period or the duration of culture increased cryosurvival, suggesting decreased lipid content due to increased β-oxidation activity. Similarly for oocyte maturation, β-oxidation appears to be able to compensate for carbohydrate deficiencies during embryo culture to some extent, and oxidation of lipids has a greater role in promoting embryo quality over increasing production rates. The findings reported in this thesis represent a contribution to the understanding of lipid metabolism during the in vitro production of porcine embryos. These results provide evidence to support a level of adaptability of porcine oocytes and embryos to different substrates available during maturation and culture. There is a preference shown for carbohydrates substrates, with the ability to utilise lipids to compensate for certain deficiencies. This would justify the inclusion of co-factors of lipid metabolism such as L-carnitine in culture media, to ensure that any deficiencies in other substrates might be corrected for and to promote higher embryo quality. Upregulation of β-oxidation also increased the cryosurvival of porcine embryos following vitrification, with this being a major development in the global movement of superior genetics for herd improvement programs. These findings will also have implications for improving in vitro culture of oocytes and embryos of other species, most notably advancements in human ARTs where research is predominantly limited to work in animal models. The understanding of how lipids are metabolised alongside exogenous carbohydrates will contribute to improving media formulations for better metabolic support in vitro, further to improving embryo production and quality.

El objetivo de la presente tesis fue estudiar qué factores pueden explicar la variabilidad existente en la congelabilidad espermática entre eyaculados, la integridad del ADN en los espermatozoides descongelados, la posibilidad de adaptar el protocolo de criopreservación a las necesidades cada verraco y, el posible papel del plasma seminal en la congelabilidad espermática. Los resultados revelaron que el verraco es el factor más importante capaz de explicar la diferente susceptibilidad entre eyaculados para soportar el proceso de criopreservación y que la variabilidad dentro de un mismo verraco es baja. El grado de fragmentación del ADN de los espermatozoides porcinos descongelados es relativamente bajo, y ligeros ajustes en la concentración de glicerol y la velocidad de descongelación, permiten mejorar la supervivencia espermática, especialmente de aquellos eyaculados considerados como "malos congeladores. Finalmente, la suplementación del medio de congelación con plasma seminal de individuos con buena congelabilidad tiene un efecto beneficioso sobre la congelabilidad espermática.
The aim of this thesis was to study which factors could explain part of the ejaculate variation in sperm freezability, the incidence of DNA fragmentation in frozen-thawed boar spermatozoa, whether different boars require modifications in the freezing and thawing protocol to enhance sperm cryosurvival and, the possible role of seminal plasma in boar sperm freezability. The results revealed that boar is the most important factor explaining the different susceptibility among ejaculates to sustain cryopreservation and that the intra-boar variability is relatively low. The overall DNA damage in frozen-thawed boar sperm is low and slight adjustment of the glycerol concentration and warming rate provides a better sperm cryosurvival, particularly in those ejaculates which are considered as "bad" freezers. Finally, supplementation of freezing media with seminal plasma from males with "good" freezability, has a beneficial effect on post-thaw sperm cryosurvival

Depuis toujours la viande représente un élément important de l'alimentation de l'homme. Cet apport alimentaire constitue le but premier de toute production domestique animale. En particulier l'industrie porcine fournit une partie très importante (environ 40%) de la production totale en viande. Le contexte économique dans lequel évolue l'industrie porcine est en constante évolution. Comme toute entreprise commerciale ayant comme objectif de maximiser son profit, une ferme porcine doit être capable d'ajuster sa stratégie de gestion rapidement. En particulier l'alimentation du bétail (la diète) et les coûts s'y rattachant doivent être contrôlés de près. Dans ce travail, nous présentons une modélisation du problème de l'engraissement porcin. Nous développons deux modèles: un premier statique et un second dynamique. Nous analysons le premier modèle à l'aide de la programmation multicritère (2 et 3 objectifs). Pour le second modèle, nous suggérons et exposons une méthode de résolution tirée du contrôle optimal et de la programmation dynamique. Finalement, nous analysons nos résultats qui, nous espérons, seront profitables à l'industrie porcine.

Dans le milieu rural malgache assez complexe et diversifie, l'elevage porcin peut se rencontrer un peu partout mais son importance varie d'une region a une autre. Il est pratique essentiellement dans les haut-plateaux et le moyenouest de madagascar et aussi dans les regions ou la migration des populations des hautes terres a ete importante. La promotion de la filiere porcine passe d'abord par une maitrise de la production agricole dont les sous-produits peuvent etre valorises par cette activite et ensuite par des meilleurs encadrement et soutien des paysans eleveurs. L'importance de la relance de cette activite dans une politique de developpement agricole repose sur son caractere complementaire aux activites agricoles des paysans et sur son role d'intermediaire qui permet a une economie de subsistance d'autoconsommation de passer a une economie d'echange. En plus, l'elevage porcin peut resoudre en partie les problemes alimentaires de la population malgache par une amelioration du regime alimentaire des producteurs et une transformation de la structure de consommation alimentaire par un apport supplementaire de proteines animales surtout dans la mesure ou la viande bovine
In the malagasy rural area, which is complicated and diversified, one can find porcine breeding everywhere, but its importance, is different from a region to another? the breeding specially takes place in the high plateau and in the middle-west of madagascar and also at the coast side of the country where the migration of the high land population is very im1ortant the promotion of the branch of the porcine breeding goes through of a control of the agricultural production whose value of the by-products can be enhanced by this activity and a better framing and then by supporting the farmers. In a politic of agricultural development, the revival of this activity is based on the fact that it comes to complete the farmers' agricultural activities and on its role of transition, which entitles an economy of subsistence of self-consumption to change into an economy of exchange. Moreover, the porcine breeding could be a solution to the food problems in madagascar by changing the structure of food consumption in supplying more animal proteins, specially when cattle-meat is

Surgical castration of piglets is a common practice in farms to avoid boar taint (androstenone and skatole) in meat, and to improve its sensory quality. The European Commission is planning to ban surgical castration by 2018 for animal welfare purposes. The aim of this thesis is to analyse the effect that a possible banning of castration would have on the Spanish pig sector. Three studies were performed: study the prevalence of boar taint in different Spanish regions; study the stakeholder’s opinion regarding the possible banning of castration; study different masking strategies for androstenone. The results showed that one out of ten entire male pigs may present high levels of boar taint, banning castration would be a problem for high quality production but not for conventional production, and breaded with garlic and parsley and fried and vacuum cooking may be useful to mask androstenone in meat.
La castració quirúrgica dels garrins és una pràctica molt utilitzada pels productors per evitar l’olor sexual (androstenona i escatol) en la carn, i per millorar-ne la qualitat sensorial. La Comissió Europea pretén eliminar la castració quirúrgica a partir de l’any 2018 per motius de benestar. L’objectiu d’aquesta tesi és analitzar l’efecte que tindria aquesta prohibició sobre el sector porcí espanyol. Es van realitzar 3 estudis: estudiar la prevalença d’olor sexual en diferents comunitats autònomes; estudiar l’opinió del sector sobre la possible prohibició de la castració; estudiar estratègies d’emmascarament d’androstenona. Els resultats mostren que 1 de cada 10 porcs mascles sencers poden presentar nivells alts d’olor sexual, que la prohibició de la castració seria problemàtica per a la producció d’alta qualitat però no per a la convencional, i que l’arrebossat amb all i julivert i fregit i la cuina al buit poden ser estratègies útils per emmascarar l’androstenona en la carn.

Dissertação de Mestrado Integrado em Medicina Veterinária
A diarreia é o sinal clínico mais frequente de presença de doença e o principal motivo de uso de antimicrobianos em leitões durante a fase de cria. No contexto da One Health, a redução de uso de antimicrobianos na produção animal é uma necessidade no combate ao desenvolvimento de resistências a antimicrobianos. Assim, o objetivo deste estudo foi avaliar o efeito da suplementação de leitões com dois extratos de algas e argila comerciais, em simultâneo, na ocorrência de diarreia neonatal, no uso consequente de antimicrobianos e o desempenho produtivo dos leitões e das ninhadas ao desmame. Para isso, ao longo de 4 bandas semanais, selecionaram-se 30 ninhadas para o grupo controlo e 36 ninhadas para o grupo tratamento, nas quais se selecionaram 7 a 8 leitões por ninhada. Tendo sido eliminados os leitões da amostra que morreram ao longo deste período, obteve-se uma amostra final de 188 leitões do grupo controlo e 177 leitões do grupo tratamento. No grupo tratamento, administrou-se oralmente Seagut® pasta a todos os leitões da ninhada nas primeiras 24h de vida e o Ecopiglet® em pó foi colocado na zona de descanso, a partir do terceiro dia de vida, durante uma semana. Os leitões foram pesados ao nascimento e na véspera do dia de desmame. Registaram-se o número de nascidos vivos, nascidos mortos e nascidos mumificados, a mortalidade, o número de leitões desmamados por porca, a ocorrência de diarreia nas ninhadas, os tratamentos antimicrobianos convencionais da ninhada completa e os tratamentos de leitões individualmente. Não se verificaram diferenças na mortalidade nem no número de leitões desmamados por porca. O ganho médio diário, por sua vez foi superior nos leitões do grupo que foi suplementado. Verificou-se uma tendência para uma diminuição da ocorrência de diarreia nas ninhadas do grupo tratado e foi possível obter uma redução significativa dos tratamentos das ninhadas completas, a par de uma tendência para uma redução dos tratamentos individuais. Em conclusão, apesar do custo superior do uso destes suplementos antimicrobianos relativamente ao uso de antimicrobianos convencionais, deve ser tido em conta que se melhorou a saúde animal e foi possível reduzir o uso de antimicrobianos.
ABSTRACT - Diarrhoea is the most frequent clinical sign of disease and the main reason for the use of antimicrobials in suckling piglets. In the context of One Health, reduction of antimicrobial use in animal production is a requirement in fighting antimicrobial resistance. Thus, the aim of this study was to evaluate the effect of piglet supplementation with two commercial algae extracts and clay products simultaneously, on the presence of neonatal diarrhoea, its consequent use of antimicrobials and the productive performance of piglets and litters. In 4 herds 30 litters were selected for the control group and 36 litters for the treatment group, in which 7 to 8 piglets per litter were selected. 188 piglets from the control group and 177 piglets from the treatment group reached the end of the experiment. In treatment group Seagut® paste was administered orally to all piglets from the litter one day postpartum and the Ecopiglet® dry powder was put in the creep area, from the third day postpartum during one week. Piglets were weighed after delivery and on day before weaning. The number of born alive, stillborn and mummified, mortality, piglets weaned per sow, occurrence of diarrhoea in litters, conventional antimicrobial treatments of the complete litter and the treatments of piglets individually were recorded. Neither mortality rate nor number of piglets weaned per sow differed between the groups. On the other hand, average daily gain was higher in supplemented piglets. There was a tendency for a decrease in presence of diarrhoea in litters in the treatment group. There was a significant reduction in treatments of the whole litters and a tendency for a reduction of the individual piglet treatments. In conclusion, despite the higher cost these antimicrobial supplements comparing to conventional antimicrobials, it should be considered animal health and welfare have been improved and there was a reduction of antimicrobial use.
N/A

Les co-infections respiratoires chez le porc sont plus fréquentes que les infections causées par un seul micro-organismes. Dans un premier temps, un recensement des études sur les co-infections respiratoires porcines a permis de mettre à jour les connaissances sur ces co-infections virales et/ou bactériennes et de détailler les probables conséquences moléculaires sur l’hôte porcin. Le virus du syndrome dysgénésique et respiratoire porcin (ou Porcine Reproductive and Respiratory Syndrome Virus, PRRSV) et les virus responsables de l’influenza porcin A (swine Influenza A Virus, swIAV), sont des acteurs majeurs du complexe respiratoire porcin. Le swIAV infecte principalement les cellules épithéliales alors que le PRRSV infecte des cellules exprimant CD163 comme les macrophages alvéolaires (MA). Dans le but d’évaluer la réponse antivirale de l'hôte porcin et d'étudier l'effet d’une pré-infection par le PRRSV sur la réplication du swIAV, une série de co-infections et de surinfections a été effectuée sur des cellules épithéliales trachéales et sur des tranches pulmonaires fines. Les résultats montrent que le PRRSV est capable d'interférer avec l'infection par swIAV et d’altérer la réponse antivirale cellulaire sans infecter les cellules épithéliales. Cet effet du PRRSV parait moins important en augmentant le délai entre les inoculations virales. Finalement, une série d’expérimentations nous a permis d’identifier les agents pathogènes circulant chez des porcs provenant d’un abattoir local et d’évaluer l’effet des différentes infections bactériennes et virales résolues ou pas sur l’immunité entraînée des macrophages alvéolaires et leur capacité à répliquer les virus suite à une surinfection. Ces travaux contribuent à la compréhension de la réponse immune porcine suite aux co-infections respiratoires, pour une meilleure gestion des maladies respiratoire chez le porc
Respiratory co-infections in pigs are more common than infections caused by a single pathogen. First of all, we identified the viral and bacterial porcine co-infections studies and we detailed the possible molecular consequences on the porcine host. The porcine reproductive and respiratory syndrome virus (PRRSV) and the swine Influenza A Virus (swIAV), are major contributors to the porcine respiratory disease complex. SwIAV primarily infects epithelial cells while PRRSV infects cells expressing CD163 such as alveolar macrophages (AM). In order to evaluate the antiviral response of the porcine host and to study the effect of a pre-infection with PRRSV on the replication of swIAV, a series of co-infections and superinfections were carried out on tracheal epithelial cells and precision-cut lung slices. The results showed that PRRSV can interfere with swIAV infection and alter the cellular antiviral response without infecting epithelial cells. This effect of PRRSV appears to be less important following an increase in the delay between viral inoculations. Finally, a series of experiments enabled us to identify the pathogens circulating in pigs from a local slaughterhouse and to assess the effect of the various bacterial and viral infections, on the alveolar macrophages trained immunity and their ability to replicate viruses in case of superinfection. This study contributes to the understanding of porcine immune response to respiratory coinfections for a better management of respiratory diseases in swine

Two types of porcine circovirus (PCV) exist, referred to as PCV1 and PCV2. PCV2 has been associated with disease syndromes in pigs, including that designated postweaning multisystemic wasting syndrome (PMWS), which has been identified in all regions of the world bar Australia (Hamel et al., 1998; Allan et al., 1999a; Onuki et al., 1999; Martelli et al., 2000; Kyriakis et al., 2000; Wellenberg et al., 2000; Done et al., 2001; Trujano et al., 2001; Saradell et al., 2004; Castro et al., 2004; Jemersic et al., 2004; Maldonado et al., 2004; Wang et al., 2004; Motovski and Segales, 2004; Garkavenko et al., 2005). PMWS affects young weaner pigs and results in weight loss, tachypnea, dyspnea, enlarged lymph nodes and jaundice (Harding, 1998). PCV2 may also cause or contribute to other swine diseases such as congential tremors (CT) (Stevenson et al., 1999), porcine dermatitis and nephropathy syndrome (PDNS) (Rosell et al., 2000), reproductive failure (Meehan et al., 2001) and several other emerging disease syndromes. PCV1 is currently considered to be non-pathogenic. Although PMWS has not been reported in Australia, information on the distribution, variation and further characterisation of PCV in Australian pigs was necessary as it might provide insights into why there is no PCV-associated disease in this country. The results reported in this thesis involved the detection and further study of porcine circovirus in Australia. This chapter provides an outline of this thesis and the work undertaken, while Chapter 2 is a review of the relevant literature with particular reference to circoviral diseases. Chapter 3 describes the detection of both PCV1 and PCV2 in the Australian pig herd, using a multiplex PCR designed to differentiate between the two viral types. The association of Australian PCV with two disease outbreaks was also investigated. Following the detection of both viruses, it was important to genetically compare Australian PCV with overseas strains known to cause disease, and this was achieved with a sequencing and phylogenetic study as described in Chapter 4. Possible reasons for the genetic groupings and distribution of different PCV2 strains worldwide are also discussed in this chapter. As PMWS is as yet unidentified in Australian pigs, the importation of pig meat into Australia from countries with the disease requires careful monitoring. Current protocols for the cooking of imported pig meat were designed to inactivate porcine reproductive and respiratory disease virus (PRRSV), and as such may not be effective against PCV. In this study (Chapter 5), Australian PCV2 was successfully infected into cell culture, and detected using a variety of techniques. Subsequently, thermal stability experiments were performed using a newly-developed immunoperoxidase (IPMA) test. It was anticipated that this study would determine whether current importation protocols require revision, and the results would suggest that this is the case, with PCV2 unaffected by treatment comparable with current cooking protocols. While no animal experiments were undertaken in this study, it may become necessary to infect pigs with Australian PCV to determine viral pathogenicity. Cell culture inoculums have been used in the past overseas, but problems with contamination and viral titre have been encountered (Fenaux et al., 2001). Viral infectious clones can be used to overcome these problems, so an infectious clone of Australian PCV2 was constructed, as described in Chapter 6. While time constraints prevented the clone from being infected into culture, it is anticipated that the construct would be infectious as it is based on a previously published method (Hattermann et al., 2004). Chapter 7 is a general discussion of the results and conclusions from this study. The detection and characterisation of Australian PCV as described in this study has provided further information on the status of PCV in the Australian pig herd, and also developed diagnostic tests to assist in future research. These tools will be important when assessing and managing the risk of Australia experiencing PCV-associated diseases.

Two types of porcine circovirus (PCV) exist, referred to as PCV1 and PCV2. PCV2 has been associated with disease syndromes in pigs, including that designated postweaning multisystemic wasting syndrome (PMWS), which has been identified in all regions of the world bar Australia (Hamel et al., 1998; Allan et al., 1999a; Onuki et al., 1999; Martelli et al., 2000; Kyriakis et al., 2000; Wellenberg et al., 2000; Done et al., 2001; Trujano et al., 2001; Saradell et al., 2004; Castro et al., 2004; Jemersic et al., 2004; Maldonado et al., 2004; Wang et al., 2004; Motovski and Segales, 2004; Garkavenko et al., 2005). PMWS affects young weaner pigs and results in weight loss, tachypnea, dyspnea, enlarged lymph nodes and jaundice (Harding, 1998). PCV2 may also cause or contribute to other swine diseases such as congential tremors (CT) (Stevenson et al., 1999), porcine dermatitis and nephropathy syndrome (PDNS) (Rosell et al., 2000), reproductive failure (Meehan et al., 2001) and several other emerging disease syndromes. PCV1 is currently considered to be non-pathogenic. Although PMWS has not been reported in Australia, information on the distribution, variation and further characterisation of PCV in Australian pigs was necessary as it might provide insights into why there is no PCV-associated disease in this country. The results reported in this thesis involved the detection and further study of porcine circovirus in Australia. This chapter provides an outline of this thesis and the work undertaken, while Chapter 2 is a review of the relevant literature with particular reference to circoviral diseases. Chapter 3 describes the detection of both PCV1 and PCV2 in the Australian pig herd, using a multiplex PCR designed to differentiate between the two viral types. The association of Australian PCV with two disease outbreaks was also investigated. Following the detection of both viruses, it was important to genetically compare Australian PCV with overseas strains known to cause disease, and this was achieved with a sequencing and phylogenetic study as described in Chapter 4. Possible reasons for the genetic groupings and distribution of different PCV2 strains worldwide are also discussed in this chapter. As PMWS is as yet unidentified in Australian pigs, the importation of pig meat into Australia from countries with the disease requires careful monitoring. Current protocols for the cooking of imported pig meat were designed to inactivate porcine reproductive and respiratory disease virus (PRRSV), and as such may not be effective against PCV. In this study (Chapter 5), Australian PCV2 was successfully infected into cell culture, and detected using a variety of techniques. Subsequently, thermal stability experiments were performed using a newly-developed immunoperoxidase (IPMA) test. It was anticipated that this study would determine whether current importation protocols require revision, and the results would suggest that this is the case, with PCV2 unaffected by treatment comparable with current cooking protocols. While no animal experiments were undertaken in this study, it may become necessary to infect pigs with Australian PCV to determine viral pathogenicity. Cell culture inoculums have been used in the past overseas, but problems with contamination and viral titre have been encountered (Fenaux et al., 2001). Viral infectious clones can be used to overcome these problems, so an infectious clone of Australian PCV2 was constructed, as described in Chapter 6. While time constraints prevented the clone from being infected into culture, it is anticipated that the construct would be infectious as it is based on a previously published method (Hattermann et al., 2004). Chapter 7 is a general discussion of the results and conclusions from this study. The detection and characterisation of Australian PCV as described in this study has provided further information on the status of PCV in the Australian pig herd, and also developed diagnostic tests to assist in future research. These tools will be important when assessing and managing the risk of Australia experiencing PCV-associated diseases.

Two types of porcine circovirus (PCV) exist, referred to as PCV1 and PCV2. PCV2 has been associated with disease syndromes in pigs, including that designated postweaning multisystemic wasting syndrome (PMWS), which has been identified in all regions of the world bar Australia (Hamel et al., 1998; Allan et al., 1999a; Onuki et al., 1999; Martelli et al., 2000; Kyriakis et al., 2000; Wellenberg et al., 2000; Done et al., 2001; Trujano et al., 2001; Saradell et al., 2004; Castro et al., 2004; Jemersic et al., 2004; Maldonado et al., 2004; Wang et al., 2004; Motovski and Segales, 2004; Garkavenko et al., 2005). PMWS affects young weaner pigs and results in weight loss, tachypnea, dyspnea, enlarged lymph nodes and jaundice (Harding, 1998). PCV2 may also cause or contribute to other swine diseases such as congential tremors (CT) (Stevenson et al., 1999), porcine dermatitis and nephropathy syndrome (PDNS) (Rosell et al., 2000), reproductive failure (Meehan et al., 2001) and several other emerging disease syndromes. PCV1 is currently considered to be non-pathogenic. Although PMWS has not been reported in Australia, information on the distribution, variation and further characterisation of PCV in Australian pigs was necessary as it might provide insights into why there is no PCV-associated disease in this country. The results reported in this thesis involved the detection and further study of porcine circovirus in Australia. This chapter provides an outline of this thesis and the work undertaken, while Chapter 2 is a review of the relevant literature with particular reference to circoviral diseases. Chapter 3 describes the detection of both PCV1 and PCV2 in the Australian pig herd, using a multiplex PCR designed to differentiate between the two viral types. The association of Australian PCV with two disease outbreaks was also investigated. Following the detection of both viruses, it was important to genetically compare Australian PCV with overseas strains known to cause disease, and this was achieved with a sequencing and phylogenetic study as described in Chapter 4. Possible reasons for the genetic groupings and distribution of different PCV2 strains worldwide are also discussed in this chapter. As PMWS is as yet unidentified in Australian pigs, the importation of pig meat into Australia from countries with the disease requires careful monitoring. Current protocols for the cooking of imported pig meat were designed to inactivate porcine reproductive and respiratory disease virus (PRRSV), and as such may not be effective against PCV. In this study (Chapter 5), Australian PCV2 was successfully infected into cell culture, and detected using a variety of techniques. Subsequently, thermal stability experiments were performed using a newly-developed immunoperoxidase (IPMA) test. It was anticipated that this study would determine whether current importation protocols require revision, and the results would suggest that this is the case, with PCV2 unaffected by treatment comparable with current cooking protocols. While no animal experiments were undertaken in this study, it may become necessary to infect pigs with Australian PCV to determine viral pathogenicity. Cell culture inoculums have been used in the past overseas, but problems with contamination and viral titre have been encountered (Fenaux et al., 2001). Viral infectious clones can be used to overcome these problems, so an infectious clone of Australian PCV2 was constructed, as described in Chapter 6. While time constraints prevented the clone from being infected into culture, it is anticipated that the construct would be infectious as it is based on a previously published method (Hattermann et al., 2004). Chapter 7 is a general discussion of the results and conclusions from this study. The detection and characterisation of Australian PCV as described in this study has provided further information on the status of PCV in the Australian pig herd, and also developed diagnostic tests to assist in future research. These tools will be important when assessing and managing the risk of Australia experiencing PCV-associated diseases.

El objetivo del presente estudio ha sido evaluar la relación entre los parámetros de la calidad seminal y la capacidad de penetración in vitro de ovocitos homólogos (PIVh) con los resultados in vivo de fertilidad y prolificidad.Se han analizado 60 eyaculados realizando sobre cada uno las medicionesclásicas del espermiograma (motilidad, calidad de movimiento, volumen, concentración espermática, estado del acrosoma, morfoanomalías y tinción vital), pruebas bioquímicas (contenido de ATP, calcio, sodio, potasio, magnesio y zinc), ensayos de la funcionalidad del espermatozoide ( test de endósmosis, test del diacetato de carboxifluoresceína, test ORT) y la valoración mediante un test de penetración in vitro homóloga. Los resultados de la valoración in vitro del semen fueron contrastados con los resultados obtenidos en un ensayo in vivo con inseminaciones homospérmicas.En nuestras condiciones experimentales el estudio de los parámetros demotilidad, morfología, acrosomas y de los test de funcionalidad de la membrana puedeser una herramienta útil, en un primer término, para desechar eyaculados de baja calidad, aunque estas técnicas no alcanzan la sensibilidad suficiente para dar un resultado satisfactorio en la predicción de la fertilidad. Sin embargo, los resultados obtenidos muestran que el test de penetración in vitro es la única técnica de las estudiadas capaz de discriminar adecuadamente los eyaculados que darán lugar a los distintos grupos de fertilidad.
The purpose of the present study was to evaluate the relationship between theparameters of seminal quality and homologous oocytes in vitro penetration capacityversus the reproductive parameters, fertility and litter size which were bothdetermined in a field trial.Sixty boar ejaculates have been analysed, performing on each of them:conventional semen analysis (motility, volume, sperm concentration, normal acrosome, morphology and structural integrity), biochemical parameters (ATP, calcium, sodium, potassium, magnesium and zinc), functional sperm assays (Hypoosmotic swelling test (HOST), Carboxiflorescin Diacetate Test (DCF), Osmotic Resistance Test (ORT)) and the homologous in vitro penetration test. The results observed upon a in vitro semen valoration were checked with those obtained upon a in vivo trial with homospermic insemination.In our experimental conditions the study of motility, morphology, normalacrosomes and functional test may be a good tool, in a first analysis, to get rid of poor seminal quality ejaculates. Still, this analysis is not accurate enough to bring outsatisfactory results to predict the in vivo fertilization capacity. In the light of theseresults only the homologous in vitro penetration test has been found able todiscriminate the different groups of fertility and litter size. None the less an extensive study is required.

This thesis explores the role of the female characters in Williams Shakespeare's play The Merchant of Venice. Through contextualizing the characters of Portia, Nerissa and Jessica within the world of early modern England, this study explores the ways in which these characters do not conform to traditional Renaissance values regarding the role of women as daughters and wives. By using historical documents such as behavioral manuals, sermons, and "defenses" of women from the late sixteenth and seventeenth century, this thesis explores the ways in which Shakespeare's female characters challenge traditional social norms. Through the comparison of the female characters with Queen Elizabeth and Patient Griselda, this study discusses the implications of the rebellious behavior of the women in The Merchant of Venice. This thesis concludes that Shakespeare purposely challenges strict social views put forward on women by creating female characters who challenge male authority and are celebrated for their behavior.

La fracture vertébrale de type burst est un traumatisme grave aux répercussions importantes pour le patient et son entourage. La définition du traitement optimal est complexe et sujette à de multiples débats au sein de la communauté médicale. Les modèles expérimentaux se sont multipliés récemment pour mieux comprendre cette fracture. Or, les modèles actuels sont limités puisqu’ils ne permettent pas d’étudier en parallèle les deux aspects fondamentaux de la fracture vertébrale : la biomécanique et les fonctions neurologiques. Ce projet vise à poser les pierres d’assises pour la création d’un modèle expérimental vivant avec une fracture vertébrale instable de type burst accompagné d’atteintes neurologiques. L’objectif de cette thèse est d’étudier la fracture vertébrale et de développer une méthode pour produire une fracture vertébrale instable in situ sur un miniporc mature complet. Trois sous-objectifs ont été définis, soit : d’étudier les mécanismes lésionnels menant à la fracture voulue, d’étudier la biomécanique des vertèbres fracturées et de concevoir l’appareil pour produire la fracture in situ. La réalisation du premier objectif a permis d’identifier les paramètres de chargement favorisant l’apparition d’une fracture vertébrale comminutive instable avec une obstruction significative du canal vertébral. Un article a été publié au mois de septembre 2011 dans le Journal of Biomechanical Egineering pour présenter les résultats obtenus (Chapitre 3). Nous avons démontré que le chargement dynamique d’un segment de colonne sans flexion favorise l’apparition de la fracture désirée. Ces caractéristiques ont été récupérées pour la conception de la méthode applicable in situ. Le second objectif visait à évaluer comment l’application d’une charge physiologique à un spécimen fracturé influence l’obstruction du canal vertébral. Les résultats obtenus ont été publiés dans le Journal of Spinal Disorders and Techniques au mois de février 2012 (Chapitre 4). Nous avons observé que l’application de charges physiologiques peut augmenter l’obstruction du canal vertébral. Cette information est importante puisque les charges intervertébrales chez les quadrupèdes sont supérieures aux charges humaines. Il faut donc considérer que l’obstruction mesurée sur les spécimens vivants sera potentiellement supérieure à ce que l’on mesure sur les spécimens cadavériques. La conception du montage permettant de produire une fracture vertébrale in situ sur un miniporc complet a ensuite été entreprise à partir des résultats obtenus au cours des étapes précédentes. L’appareil a été fabriqué et les premières fractures obtenues sont décrites dans un article soumis a The Spine Journal au mois de mai 2012 (Chapitre 5). Des fractures comminutives instables avec une obstruction significative du canal vertébral ont été produites in situ sur des spécimens complets. La méthode développée n’a pas causé de fractures additionnelles aux structures adjacentes. La méthode et l’appareil développés dans le cadre de cette thèse sont sans précédent. Cette méthode produit des fractures d’intérêts pour la communauté scientifique et localise le traumatisme au rachis. Avec quelques modifications à l’équipement, un modèle expérimental vivant avec une fracture instable de type burst et des atteintes neurologiques potentielles pourra être réalisé. Ce modèle pourra être utilisé dans le cadre de plusieurs études exploratoires et précliniques.

Le porc est la seule espece de mammifere connue ou le trophectoderme de l'embryon, en cours d'implantation, secrete de fortes quantites d'interferon-gamma (ifn-), en meme temps qu'un autre ifn, l'ifn-delta. Nous avons tout d'abord etudie la structure et les proprietes biochimiques de cet ifn embryonnaire, en les comparant a celles de l'ifn- leucocytaire adulte. Du fait de son origine epitheliale, l'ifn- trophoblastique differe de l'ifn- leucocytaire par une structure polypeptidique tronquee, par le nombre de glycoformes (deux au lieu de quatre), et par le niveau de glycosylation des monomeres. Ces differences structurales entrainent des proprietes biochimiques specifiques concernant le point isoionique (pl), la stabilite a differents ph, et la capacite a se replier apres denaturation. Dans une seconde partie nous nous sommes interesses aux effets biologiques des deux ifn embryonnaires sur l'uterus gestant. Par coloration immunohistochimique de coupes d'uterus, nous avons pu montrer que l'ifn-gamma traverse l'epithelium endometrial, et penetre dans le stroma uterin, ou il induit fortement les antigenes d'histocompatibilite de classe ii, en particulier sur les endotheliums vasculaires. Dans un modele experimental utilisant deux lignees cellulaires epitheliales (luminale et glandulaire), cultivees sur membranes microporeuses, nous avons montre que l'ifn-delta est l'agent principal sinon exclusif d'une depolarisation de l'epithelium luminal. Nous concluons que l'ifn- a pour cible la muqueuse uterine (stroma, edotheliums, glandes), contribuant probablement a creer un environnement non-infectieux pour le conceptus. Le role principal de l'ifn- serait alors de preparer l'uterus a l'implantation et/ou de permettre a l'ifn- de traverser la barriere epitheliale.

Le but de cette étude était d'évaluer le potentiel antimicrobien de bactériocines pour le contrôle du pathogène porcin Streptococcus suis sérotype 2. Pour répondre au premier objectif de ce projet, la susceptibilité de S. suis à la nisine, une bactériocine produite par Lactococcus lactis, seule ainsi qu'en combinaison avec différents antibiotiques, a été démontré. Pour réaliser le second objectif, la purification et la caractérisation d'une bactériocine produite par une souche non-virulente de S. suis sérotype 2 ont été effectuées. La bactériocine a d'abord été purifiée par HPLC pour ensuite être caractérisée au niveau biochimique, ainsi que génique. Des études plus approfondies permettront d'évaluer la capacité de la nisine, de la suicine 90-1330 ou de leur souche productrice respective à prévenir des infections expérimentales à S. suis chez le porc en vue d'une potentielle application thérapeutique et préventive.

Durante años se ha tratado de simular in vitro el proceso de selección y capacitación espermática que lleva a cabo el tracto genital de la hembra en condiciones fisiológicas, pero los resultados mucho de la selección que ocurre en el oviducto in vivo. El objetivo principal de esta tesis fue: el estudio de las subpoblaciones espermáticas previamente seleccionadas a través de métodos in vitro (por gradientes de Percoll, Capítulo 1; en cultivo de células oviductales, Capítulo 2; y la combinación de ambos sistemas, Capítulo 3), ex vivo (en explantes oviductales, Capítulo 2) e in vivo (en el oviducto de la cerda tras una inseminación artificial, Capítulo 2), y su relación con la funcionalidad y el estado de capacitación, así como su capacidad fecundante. En el Capítulo 1 se analizó la centrifugación mediante gradientes de Percoll como método de selección y capacitación espermática y se evaluó la eficacia en la selección de espermatozoides en función del gradiente utilizado (45/60, 60/75 y 45/90%). En el experimento 1, se analizaron diferentes parámetros espermáticos de las poblaciones obtenidas a través de los 3 grupos experimentales. Los resultados demostraron que al aumentar la diferencia en la densidad del gradiente de Percoll se reduce el número de espermatozoides morfológicamente anormales o con fragmentación del ADN, recuperando principalmente, espermatozoides que han iniciado el proceso de capacitación (95% presentan fosforilación de tirosina en alguna zona). Podemos decir que la separación de espermatozoides mediante gradientes de Percoll se fundamenta en sus diferencias funcionales. En el experimento 2, se emplearon las poblaciones espermáticas en fecundación in vitro para evaluar su capacidad de penetración, y determinar qué gradiente selecciona los espermatozoides más aptos para fecundar y su posible relación con los parámetros estudiados en el experimento 1. Se evaluó el porcentaje de penetración y el número medio de espermatozoides que presentaba cada ovocito. Los resultados mostraron que aquellos espermatozoides obtenidos a partir del gradiente de Percoll más restrictivo (45/90%) dieron lugar a unos mayores niveles de penetración del ovocito. En el Capítulo 2 se estudió el efecto de las células epiteliales del oviducto sobre la capacitación espermática. Se evaluó el estado de capacitación (mediante la distribución de la fosforilación de la tirosina) de los espermatozoides seleccionados mediante células oviductales bajo tres condiciones experimentales: in vitro, ex vivo e in vivo. Los resultados mostraron que existen diferencias entre los espermatozoides unidos o no unidos a las células oviductales, lo que demuestra que este tipo de células seleccionan espermatozoides con bajo grado de fosforilación en tirosina en cualquiera de las condiciones estudiadas. En el Capítulo 3 se estudiaron qué características presentan los espermatozoides para ser seleccionados por las células oviductales y la capacidad de fecundación in vitro de estos espermatozoides doblemente seleccionados [a través de gradientes de Percoll (45/90%) y posteriormente incubados sobre cultivos in vitro de células epiteliales del oviducto]. En el experimento 1 se evaluó la capacidad para fecundar de las dos poblaciones de espermatozoides obtenidas mediante el análisis de diferentes parámetros de fecundación. Los resultados demostraron que los espermatozoides doblemente seleccionados dan mejores resultados de fecundación in vitro, mejorando las tasas de penetración del gameto femenino. En el experimento 2 se analizaron las características funcionales de los espermatozoides. Los resultados indicaron que las células oviductales unen espermatozoides de mayor calidad y realizan su propia selección incluso partiendo de espermatozoides previamente seleccionados. En resumen, los resultados de esta tesis doctoral muestran que las células epiteliales del oviducto son capaces de distinguir aquellos espermatozoides con una capacidad de fecundación superior, lo que en un futuro podría representar un incremento en la eficiencia de los sistemas de reproducción asistida evitando, de este modo, la selección parcialmente subjetiva que existe actualmente en las técnicas convencionales de selección espermática.
For many years, attempts have been made to control sperm selection and capacitation in order to simulate under in vitro conditions, but the results are highly variable and are far from in vivo ones. The aim of this thesis was the study of sperm subpopulations that have been previously selected by different methods: in vitro (Percoll gradients in Chapter 1, oviductal cells culture in Chapter 2 and by combining both in Chapter 3), ex vivo (in oviductal explants, Chapter 2) and in vivo (in the oviduct of the sow after insemination, Chapter 2), and their relationship with functionality, sperm capacitation status and fertilizing capacity. In Chapter 1 centrifugation by Percoll gradients was analysed as a sperm selection and capacitation method, and its efficiency for selecting sperm was evaluated. Three different Percoll gradient combinations (45/60, 60/75 and 45/90%) were used. In experiment 1, several parameters related to sperm function were analysed in the different sperm populations obtained through the three experimental techniques. The results of this experiment showed that as the Percoll gradient increased, the number of morphologically abnormal sperm or sperm with DNA fragmentation is reduced, recovering especially those sperm that have begun the process of sperm capacitation (since 95% of spermatozoa present tyrosine phosphorylation in any of the studied areas). So, we can say that the separation of sperm by Percoll gradients is primarily based on their functional differences. In experiment 2, the different sperm populations were used in an in vitro fertilization system to evaluate the penetration rate and checking which Percoll gradient selects the best sperm for fertilization purposes. The penetration rate and the mean number of spermatozoa found in each oocyte were evaluated. The results showed that spermatozoa obtained from the most restrictive Percoll gradient (45/90%) had higher levels of penetration. In Chapter 2 the capacitation status of sperm selected by the oviductal epithelial cells was evaluated (by measuring levels of tyrosine phosphorylation) under three conditions: in vitro, ex vivo and in vivo. The results showed that the phosphorylation pattern changes when sperm are incubated with the epithelial cells of the oviduct, with differences between sperm bound or unbound to the cells. It was concluded that the epithelial cells of the oviduct are able to select spermatozoa with a low level of tyrosine phosphorylation in any of the studied conditions. In Chapter 3 sperm selected by Percoll gradients (45/90%) were incubated with in vitro cultured oviductal epithelial cells. This incubation resulted in two distinct sperm populations: sperm bound to cells and sperm unbound. The study was divided into two experiments. In experiment 1 the ability of the two populations to fertilize was evaluated using different parameters. The results showed that the doubly selected sperm led to better results in the in vitro fertilization systems, significantly improving penetration rates. In experiment 2 the functional characteristics of sperm from the two populations were analysed. The results indicated that the oviductal epithelial cells bind higher quality sperm according to different functional parameters and perform their own sperm selection. In summary, the results of this thesis show that the oviductal epithelial cells are able to distinguish those sperm with a higher fertilization capacity. In the future this could provide an increase in the efficiency of assisted reproduction systems thereby preventing the partially subjective selection that currently exists in conventional sperm selection techniques.

El cerdo es una de las principales fuentes de carne consumida por el hombre y las preferencias de los consumidores hacia productos de alta calidad han aumentado durante los últimos años. Por lo tanto, conocer los mecanismos moleculares que afectan a la producción y a la calidad de esta carne ayudaría a la selección de estos caracteres. La calidad de la carne está determinada en gran medida por la composición de los ácidos grasos (AG) y la comprensión de los procesos moleculares subyacentes a éstos son el objetivo general de esta tesis. En este trabajo, se han identificado QTLs en el cromosoma 8 porcino (SSC8) para la composición de AG en grasa dorsal (GD) y se han identificado dos regiones cromosómicas con SNPs asociados, localizadas a 93 y 119 Mb. Las señales estadísticamente más significativas para ambas regiones se observaron para el ácido palmitoleico y los índices C18:0/C16:0 y C18:1(n-7)/C16:1(n-7). Los genes MAML3 y SETD7 fueron estudiados como genes candidatos posicionales para la región localizada a 93 Mb. Los dos nuevos microsatélites analizados en el gen MAML3 y el SNP del gen SETD7 (SETD7:c.700G>T) no mostraron las asociaciones más significativas en esta región, descartando estos polimorfismos como las mutaciones causales. Además, en la región localizada a 119 Mb, el SNP ELOVL6:c.-533C>T mostró la asociación más significativa con el porcentaje de los ácidos palmítico y palmitoleico y los índices de elongación en GD. Los resultados obtenidos para el gen ELOVL6, gen candidato posicional del QTL localizado a 119 Mb refuerzan la hipótesis de su efecto pleiotrópico sobre la composición de AG en GD y en músculo, y su papel en la determinación del QTL del SSC8 para el perfil de AG. Por otra parte, se utilizaron datos del genoma completo de cerdos ibéricos y landrace para identificar 1.279 variaciones en el número de copias (CNV), las cuales se fusionaron en 540 regiones de CNVs (CNVRs). El impacto de cuatro de ellas fue estudiado para caracteres de crecimiento y composición de AG. Se encontró asociación con la longitud de la canal y la composición de AG en grasa intramuscular y GD para el CNVR112. Este CNVR contiene el gen GPAT2 que cataliza la biosíntesis de triglicéridos y glicerofosfolípidos. Los resultados obtenidos subrayan la importancia de los CNVRs en la determinación de caracteres económicamente importantes en el cerdo. Finalmente, se analizó la expresión de 44 genes candidatos relacionados con el metabolismo lipídico en 115 animales. El estudio de asociación genómico con los datos de expresión (eGWAS) identificó 193 eSNPs localizados en 19 eQTLs. Tres de los eQTLs correspondientes a los genes ACSM5, FABP4 y FADS2 se clasificaron como cis-eQTLs; mientras que los 16 eQTLs restantes mostraron efectos reguladores en trans. Estos hallazgos, junto con los polimorfismos evaluados para alguno de estos genes, mejoran nuestro conocimiento sobre los mecanismos funcionales implicados en la variación de los caracteres relacionados con la calidad de la carne porcina.
Pork is one of the main sources of human-consumed meat and consumer’s preference towards high quality meat is increasing. Hence, understanding the molecular mechanisms affecting meat production and quality would help in the selection of these traits. Meat quality is determined largely by its fatty acid (FA) composition and understanding the underlying molecular processes of FA composition is the general objective of this thesis. We analyzed quantitative trait loci (QTL) on porcine chromosome 8 (SCC8) for FA composition in backfat, identifying two trait-associated SNP regions at 93 Mb and 119 Mb. The strongest statistical signals for both regions were observed for palmitoleic acid and, C18:0/C16:0 and C18:1(n-7)/C16:1(n-7) elongation ratios. MAML3 and SETD7 genes were analyzed as positional candidate genes in the 93 Mb region. The two novel microsatellites analyzed in the MAML3 gene, and the SETD7:c.700G>T SNP in the SETD7 gene did not show the strongest signal in this region, discarding these polymorphisms as the causal mutations. Furthermore, in the 119 Mb region, the ELOVL6:c.-533C>T SNP showed a strong association with the percentage of palmitic and palmitoleic acids and elongation ratios in backfat. These results for ELOVL6 gene, support the hypothesis that it has a pleiotropic effect in backfat and muscle for the 119 Mb QTL, and reinforce this gene as a strong candidate for the SSC8 QTL for FA composition. Moreover, whole genome sequence (WGS) data from Iberian and Landrace pigs were used to identify 1,279 copy number variations (CNVs), merging into 540 swine CNV regions (CNVRs). The impact of four of them in growth and FA composition in intramuscular fat and backfat was studied. Association with carcass length and FA composition in backfat and intramuscular fat was showed for the CNVR112, containing the GPAT2 gene which catalyse the biosynthesis of triglycerides and glycerophospholipids. These results underline the importance of CNVRs affecting economically important traits in pigs. Finally, the adipose tissue mRNA expression of 44 candidate genes related with lipid metabolism was analyzed in 115 animals. The expression genome-wide association (eGWAS) identified 193 eSNPs located in 19 expression QTLs (eQTLs). Three out of 19 eQTLs corresponding to ACSM5, FABP4, and FADS2 were classified as cis-acting eQTLs, whereas the remaining 16 eQTLs had trans-regulatory effects. These findings and the polymorphisms evaluated for some of these genes provide new data to further understand the functional mechanisms implicated in the variation of meat quality traits in pigs.

Mestrado em Engenharia de Materiais
O trabalho realizado tem como principal objectivo obter cerâmicos tricamada porosa/densa/porosa de YSZ (sigla inglesa para Yttria-Stabilized Zirconia), com micro e macroporosidade de geometria ajustável, recorrendo a réplicas de materiais orgânicos fibrosos e particulados, bem como obter células electroquímicas de alta temperatura por inflitração de soluções de precursores adequados nas camadas porosas de YSZ. Recorreu-se a várias técnicas de caracterização tais como o cálculo de densidades aparentes pela relação volume:massa e pela espessura das diferentes camadas, Método de Arquimedes, Microscopia Electrónica de Varrimento, Análises Térmicas de ATD/TG e Difracção de Raios X. Foram estudadas diversas variáveis nomeadamente, escolha e selecção de agentes porogéneos – fibrosos e particulados –, composição da mistura que dará origem às camadas porosas, isto é, equilíbrio entre o volume incorporado de agente porogéneo e de pó cerâmico, bem como a relação entre o agente fibroso e o particulado; testes para seleccionar entre o ciclo de sinterização convencional e o ciclo de sinterização em duas etapas e eventuais melhorias no processamento cerâmico. Os agentes porogéneos seleccionados foram as fibras de celulose enquanto agente fibroso, e amido de milho enquanto agente particulado. Relativamente à composição, veio a provar-se dos testes efectuados que aquela que mais se adequa e melhores resultados apresentou apresenta 70% de agente porogéneo e 30% de pó cerâmico, 3-YSZ. Quanto à relação entre os dois agentes porogéneos na mistura, veio a evidenciar-se a necessidade de incorporar uma quantidade substancialmente maior de fibras de celulose, 70 - 80% e, 20 a 30% de amido de milho. A co-sinterização de multicamadas cerâmicas, com e sem agentes porogéneos, recorrendo a ciclos em duas etapas permitiu obter estruturas multicamada densa-porosa em que as camadas porosas apresentam, num corpo mecanicamente estável, simultaneamente macroporosidade e nanoporosidade, enquanto que as camadas densas apresentam níveis de densificação superiores a 95%. ABSTRACT: The main goal of this work was to obtain triple-layered ceramics porous/dense/porous of YSZ (Yttria-Stabilized Zirconia), with micro and macroporosity of adjustable geometry, using fibrous and particles organic materials, as also obtain high temperature electrochemical cells by impregnation of appropriate precursors solutions on the porous layers of YSZ. Several characterization techniques were used in particular the determination of the apparent density by the relationship between volume:mass and the thickness of the different layer, Archimedes’s Method, Scanning Electronic Microscopy, Thermal Analysis of DTA/TG and X Ray Diffraction. Several variables were studied like the choice and selection of the materials – fibrous and particulate – in order to create porosity; composition of the mixture that will become the porous layers, the relation between organic elements and ceramic powder, as well as the combination between the fibrous and particulate agent; selection of the best sintering cycle – conventional cycle or two step sintering cycle –, and also improvements in ceramic processing. The organic materials selected were cellulose fibers as fibrous element, and corn starch as particulate element. About the composition, the results showed that the best was 70% of porous former element, and 30% of ceramic powder, 3-YSZ. The relation between the two organic elements was 75% in volume for cellulose fibers, and 25% for corn starch. The results showed that it was necessary to incorporate much more volume of fibers than starch. The sintering of ceramic, with and without porous former, in two steps seems to offer better results in densification of dense layers, and also The co-sintering of multilayer ceramics, with and without pore formers, using cycles in two steps, allow to obtain dense-porous multilayered structures, where porous layers were mechanically stable, with macro- and nanoporosity, while the dense layers present levels of densification above 95%.