Muestras de músculo esquelético y sangre fueron tomadas a 48 toros bravos (novillos) lidiados en la plaza de toros de Calasparra (Murcia). Los músculos seleccionados, cabeza larga del tríceps braquial y semitendinoso, fueron analizados desde el punto de vista histológico, con el fin de estudiar las características morfológicas e histoquímicas de los distintos tipos de miocitos y las posibles alteraciones consecuencia de la lidia. Las técnicas histoquímicas han posibilitado reconocer tres tipos de fibras musculares, denominadas I (SO), IIA (FOG) y IIX (FG). El músculo tríceps braquial presentó un porcentaje mayor de fibras oxidativas (SO + FOG) en comparación con el músculo semitendinoso donde predominaron las fibras glicolíticas (FG). El estudio histológico del músculo esquelético reveló además que tras la lidia algunas fibras musculares sufrieron lesiones agudas de tipo degenerativo. Estas lesiones se presentaron de forma aislada, afectando a las fibras de tipo II, sin que se viera comprometida la integridad funcional de los fascículos musculares. En un porcentaje reducido de animales se observaron lesiones crónicas relacionadas con la presencia de fibras anguladas atróficas y quistes de Sarcocystis spp. Las muestras de sangre fueron procesadas con objeto de valorar diversas enzimas indicativas del posible daño renal, hepático y muscular consecuencia de la lidia. Los valores de enzimas estudiados en sangre demuestran que en la mayoría de los novillos la lidia no produjo daño renal, ni hepático, ni muscular. Tan solo 5 ejemplares presentaron niveles algo más elevados de lo normal en las enzimas LDH y CK que no se correspondieron con un mayor grado de lesión muscular. Este trabajo demuestra que el músculo esquelético de los novillos estudiados se encontraba bien preparado para afrontar el esfuerzo que supone la lidia. / The samples were taken from blood and skeletal muscle in 48 fighting bulls at bullring in Calasparra (Murcia). Selected muscles, long head of the triceps brachial and semitendinosus were analyzed histologically, in order to study morphological and histochemical characteristics of different types of myocytes and their possible alterations as consequence of the fighting. Three types of muscle fibers, called I (SO), IIA (FOG) and IIX (FG) were identified from histochemical techniques. The triceps brachii muscle presented a higher percentage of oxidative fibers (SO + FOG) when comparing with semitendinosus where glycolytic fibers (FG) were highest. Histological examination of skeletal muscle also revealed that after fighting some muscle fibers suffered acute degenerative injuries. These injuries were scarce, such that these only affected to the type II fibers, without affecting the functional integrity of the muscle bundles. A small percentage of animals showed chronic injuries corresponding with atrophic angulated fibers and cysts of Sarcocystis spp. Blood samples were processed in order to assess several enzymes indicative of kidney damage, liver damage and muscle damage as result of the fight. The enzymes values in blood showed that the fight did not cause damages in kidney, liver, and muscle in the most of the bulls. Only 5 bulls showed levels of LDH and CK enzymes higher than the normal levels without observing positive correlation with the degree of muscle injury. This work shows that the skeletal muscle of the bulls was well prepared to face the effort involved in the fight.

Un área muy actual de investigación es el estudio de las causas del envejecimiento. En el ámbito de la biología reproductiva también lo está siendo por sus implicaciones tanto en la medicina como en la veterinaria. Si durante muchos años los estudios se han centrado en la mujer o en las hembras para explicar el descenso reproductivo con la edad actualmente se está prestando más atención al factor masculino. Estos estudios abordan varios aspectos: cambios endocrinos, alteraciones de los espermatozoides, trasmisión de enfermedades hereditarias, o la necesidad de terapias sustitutivas de testosterona en la vejez. Respecto a los cambios histológicos del testículo envejecido, se han realizado diversos estudios desde hace décadas, pero han dejado ciertas cuestiones sin aclarar y se han centrado en el hombre y en la rata. Junto a esto, durante los últimos años, diversos autores han propuesto que el mecanismo tisular implicado en la progresiva disminución de la espermatogénesis con la edad, está relacionado con cambios en el equilibrio entre la proliferación/apoptosis en el epitelio seminífero. En la presente tesis doctoral nos hemos propuesto por un lado, realizar un completo análisis histológico tanto cualitativo como cuantitativo de los cambios que acontecen con la edad en el epitelio seminífero y pared tubular del hámster sirio. Por otro, hemos investigado también si la proliferación está o no alterada en este epitelio relacionándola con las alteraciones histológicas observadas en el mismo. Para ello, utilizamos hámsteres machos de 6, 12 y 24 meses de edad. Los testículos fueron fijados y procesados para microscopia de luz convencional e inmunohistoquimica y, electrónica de transmisión. Se realizó un estudio semicuantitativo del tipo histológico de degeneración de las secciones de los túbulos seminíferos determinando el grado de progresiva alteración de su epitelio con la edad. Inmunohistoquímicamente se identificó la inmunoreactividad a laminina y fibronectina en la pared tubular y las células en proliferación. Morfométricamente se calculó una serie de parámetros testiculares y el volumen total de inmunoreactividad a fibronectina y laminina. Respecto a la proliferación se calculó el índice total de proliferación como el número de células proliferando por tipo histológico de sección tubular. Por último, con las secciones ultrafinas se realizó una descripción ultrarestructural del epitelio seminífero y se calculó el grosor de la pared tubular. En los animales envejecidos se observó un incremento significativo de las secciones tubulares con diversas alteraciones del epitelio seminífero especialmente hipoespermatogénesis y detención de la espermatogénesis. El volumen del testículo, el diámetro y volumen del túbulo como el volumen de epitelio del túbulo seminífero sufrieron un descenso significativo con la edad, por el contario la longitud del túbulo se incrementó. El volumen de fibronectina y laminina aumento en los animales viejos siendo el índice de proliferación menor en estos animales. El número de células en proliferación por tipo histológico de sección de túbulo seminífero fue progresivamente y significativamente menor en las secciones alteradas respecto a las secciones normales. No se observó diferencias significativas respecto a la edad entre tipos histológicos de sección de túbulo seminífero. Ultraestructuralmente se observó alteraciones citológicas importantes en la célula de Sertoli así como en el proceso de espermiogénesis. La pared presentó un engrosamiento significativo tanto en los túbulos normales como en los hipoespermatogénicos o en detención de la espermatogénesis. En conclusión, el envejecimiento del túbulo seminífero del hámster supone una pérdida progresiva de la actividad espermatogénica que está relacionada con cambios del túbulo y de su pared. Estos cambios no se producen de forma uniforme en todo él y se asocian a una disminución de la proliferación celular en sus zonas alteradas histológicamente. Éstas presentan además células de Sertoli modificadas ultraestructuralmente. / A very current area of research is the study of the causes of aging. In the field of reproductive biology the aging is important by their implications both in medicine and veterinary. If for many years the studies have focused on women or females to explain the reproductive decline with age it is now being paid more attention to male factor. These studies address several aspects: endocrine changes, abnormal sperm, transmission of hereditary diseases, or the need for testosterone replacement therapy in older age. Regarding the histological changes of aging testes, they have conducted several studies for decades, but have left some issues unclear and have focused on humans and rats. Alongside this, in recent years, several authors have proposed that the mechanism involved in decrease of spermatogenesis with age, is related to changes in the balance between proliferation / apoptosis in the seminiferous epithelium. In this thesis we have proposed on the one hand, a complete histological analysis of qualitative and quantitative changes that occur with age in the seminiferous epithelium and tubular wall of the Syrian hamster. Furthermore, we have also investigated whether this proliferation in the epithelium altered is linked with the histological changes observed in it. To do this, we use male hamsters 6, 12 and 24 months old. The testes were fixed and processed for conventional light microscopy, immunohistochemistry and transmission electron microscopy. A semiquantitative histological study of sections of seminiferous tubules for determining the degree of gradual deterioration with age, in its epithelium was performed with age. Immunoreactivity to laminin and fibronectin in the tubular wall and proliferating cells was identified. Morphometrically a series of testicular parameters and the total volume of fibronectin and laminin immunoreactivity was calculated. The total proliferation index as the number of proliferating cells by histologic type of tubular section was calculated. Finally, with ultrathin sections, an ultrarestructural description of seminiferous epithelium, and thickness determination of tubular wall was performed. In aged animals a significant increase in the tubular sections with various disorders especially hypospermatogenesis of seminiferous epithelium and arrest of spermatogenesis was observed. The testicular volume, diameter and volume as epithelial volume of seminiferous tubule suffered a significant decrease with age, on the contrary an increased tubule length was observed. An increase in fibronectin and laminin volume in old animals and less proliferation rate in these animals was determined. The number of proliferating cells per histologic section of seminiferous type tubule, progressively and significantly decrease in altered sections respect to normal sections. No significant differences respect to age between histological types sections of seminiferous tubule was observed. Ultrastructurally important cytological changes were observed in the Sertoli cell and the process of spermiogenesis. The wall thickening showed an increase significant in normal, hipospermatogenic and spermatogenesis arrest tubules. The number of proliferating cells per histologic section of seminiferous type tubule, progressively and significantly decrease in altered sections respect to normal sections. In conclusion, the aging of the seminiferous tubule hamster suppose a loss of spermatogenic activity which is related to changes in the tubule and its wall. These changes do not occur uniformly throughout the tubule and they associated with a decrease in cell proliferation in altered tubular areas. These areas also have Sertoli cells with a modified ultrastructure.

Objetivos 1. Describir los abordajes ecográficos para la evaluación del plexo braquial (PB) y los principales nervios del miembro torácico, así como la anatomía y apariencia ecográfica normales correlacionando las imágenes ecográficas con la disección anatómica y las criosecciones. 2. Establecer los abordajes ecográficos para el bloqueo ecoguiado del PB. 3. Determinar la eficacia de los diferentes abordajes para el bloqueo anestésico ecoguiado del PB, mediante la evaluación de la distribución de tinta o contraste iodado inyectado alrededor de los nervios de interés. 4. Evaluar clínicamente la eficacia del bloqueo ecoguiado del PB. Material y métodos En la fase in vitro del estudio se emplearon 37 cadáveres de gatos adultos. Todos los animales fueron eutanasiados de forma humanitaria por razones no relacionadas con el presente estudio. En la fase in vivo se utilizaron trece gatos experimentales. Los exámenes ecográficos se efectuaron utilizando una sonda lineal de 4-13 MHz. Todos los exámenes ecográficos in vitro se realizaron inmediatamente después de la eutanasia. En el estudio anatómico se realizaron disecciones del PB y sus principales componentes en cuatro cadáveres. Otros cuatro cadáveres se congelaron durante 48 horas, y posteriormente, se realizaron criosecciones desde la columna vertebral hasta el codo. El examen ecográfico del PB se realizó en cinco cadáveres. El PB fue en dos zonas: axilar y humeral. Se evaluó la apariencia, ecogenicidad y ecotextura de los nervios evaluados. En el estudio “in vivo”, la evaluación ecográfica del PB se llevó a cabo en cinco gatos. El protocolo ecográfico utilizado para esta parte del estudio fue el mismo que el ya descrito en la parte anterior. Para determinar la eficacia de dos abordajes ecográficos diferentes se emplearon un total de 24 cadáveres. Los animales fueron divididos en dos grupos: (1) se posicionaron en decúbito dorsal con los miembros torácicos flexionados y orientados caudalmente (técnica FAD), (2) se posicionaron con el miembro torácico que iba a ser bloqueado en abducción 90º y el contralateral orientado caudalmente (técnica FAB). Una vez identificadas las raíces nerviosas del PB mediante ecografía, se insertó una aguja para bloqueo de nervios periféricos utilizando una técnica en plano y se inyectó 1 ml de una solución de tinta azul. Para el estudio de TC, se inyectó 1 ml de medio de contraste iodado, utilizando la misma técnica. Para la evaluación clínica de la eficacia del bloqueo ecoguiado del PB emplearon ocho gatos adultos. Los animales fueron colocados en decúbito dorsal con el miembro torácico que iba a ser bloqueado en abducción 90º y el contralateral orientado caudalmente. Una vez identificadas las raíces del PB, se insertó una aguja para el bloqueo de nervios periféricos utilizando una técnica en plano y se inyectó lidocaína al 1%. El éxito del bloqueo se determinó mediante la evaluación de la función sensorial y motora de la extremidad bloqueada. Conclusiones 1. Los abordajes para el estudio ecográfico del PB y sus componentes incluyeron diferentes zonas: axilar, humeral medial (a tres niveles: proximal, medio, distal) y humeral lateral (a nivel distal). 2. El aspecto ecográfico del PB es el de un cluster de estructuras redondeadas que se corresponden con las raíces nerviosas C6, C7, C8 y T1. En el abordaje humeral medial se visualizaron los nervios radial, mediano y cubital, y en el abordaje humeral lateral se observó la rama superficial del nervio radial. 3. La distribución de la tinta y del contraste iodado permitió establecer la eficacia de los abordajes evaluados para el bloqueo del PB. 4. El abordaje axilar con la extremidad a bloquear abducida 90º y la contralateral dirigida caudalmente se consideró óptimo para realizar el bloqueo ecoguiado del plexo braquial en el gato. / Objectives 1. To describe ultrasonographic approaches for scanning the brachial plexus (BP) and major nerves of the thoracic limb in the cat. To describe and the normal ultrasonographic anatomy and appearence of these nerve structures, and to correlate with the corresponding anatomical dissesctions and cryosections. 2. To describe ultrasonographic approaches for the ultrasound guided axillary BP block . 3. To assess the usefulness of these ultrasonographic approaches for the blockade of the BP, by means of injecting ink or contrast around the target nerves. 4. To evaluate the clinical efficacy of the ultrasound guided axilary brachial plexus block. Material and methods The in vitro study was performed in 37 fresh adult feline cadavers. The cadavers were humanely euthanized for reasons unrelated to thoracic limb lameness on the day of the study. Thirteen adult healthy experimental cats were used to perform the in vivo study. A 4–13 MHz linear array transducer was used for all the examinations. Anatomic dissections of the BP and major components were performed in 4 fresh feline cadavers. Another four cadavers were frozen for 48 hours, and transverse cryosections of the front limb were performed from the spine to the elbow. Ultrasonographic examination of BP was performed five fresh cadavers. The BP was scanned in two zones: axillary and humeral. The appearance, echogenicity and echotexture of components of the BP, major nerves, and surrounding structures were recorded. The “in vivo” ultrasonographic evaluation of the BP was carried out in five healthy experimental cats. The US protocol used for this study was the same as the one described in the second study above. To assess the usefulness of these ultrasonographic approaches for the blockade of the BP a total of 24 fresh cadaver cats were employed . The animals were divided into two groups: (1) cats were placed in dorsal recumbency with both forelimbs adducted (thoracic limbs flexed and orientated caudally) (FAD technique), (2) cats were positioned with the forelimb to be blocked abducted 90° and the other one adducted to the side (FAB technique). Once the nerve roots of the BP were identified by US, a peripheral nerve block needle was inserted in-plane with respect to the ultrasound transducer, and 1 ml of dye solution was injected. For the CT study, 1 ml of iodinated contrast medium was injected as described above. To evaluate the clinical efficacy of the ultrasound guided axilary brachial plexus block 8 adult experimental male cats were employed. The animals were placed in dorsal recumbency with the limb to be blocked abducted 90º. A peripheral nerve block stimulating needle was inserted in–plane with respect to the ultrasound transducer and Lidocaine 1% was injected. The success of the BP blockade was determined by the assessment of the motor and sensory function. Conclusions 1. The approaches for the scanning the BP and its components include: axillary, medial humeral (at three levels: proximal, middle, distal) and lateral humeral (distal). 2. The sonographic appearance of BP at the axillary level on transverse plane was a cluster of oval to rounded structures that correspond to nerve roots C6, C7, C8 and T1. In the medial humeral approach, the radial, median and ulnar nerves were displayed, and in the lateral humeral approach the superficial branch of the radial nerve was despicted. 3. The pattern of distribution of the ink and contrast allowed to establish the efficacy of the ultrasonographic approaches developed in this study for the blockade of the BP. 4. The axillary approach with the limb to be block abducted 90º and the contralateral limb directed caudally was considered optimal for performing ultrasound-guided brachial plexus block in the cat.

El uso de muestras seminales criopreservadas es muy habitual en la reproducción asistida humana y de animales con interés en ganadería, así como en la conservación de razas y especies en peligro de extinción. Sin embargo, la congelación y descongelación de muestras seminales implica alteraciones en la membrana y en el ADN del espermatozoide, procesos de apoptosis y modificaciones asociadas al estrés oxidativo (Polge y Jakobsen, 1959; Pursel y Johnson, 1975; Holt, 2000; Thomson et al. 2009). Los espermatozoides y las células del sistema reproductor masculino presentan sistemas de defensa frente a los radicales libres (Sies, 1993). En este contexto, el glutatión es uno de los antioxidantes más ubicuos e importantes en los sistemas biológicos (Gadea et al., 2005). Sin embargo, en ocasiones tiene lugar una producción excesiva de radicales libres, un detrimento en la capacidad total antioxidante o ambas situaciones a la vez. Esta circunstancia implica un desbalance denominado “estrés oxidativo” (Sies y Cadenas, 1985). La adición de antioxidantes a los medios de congelación y descongelación seminal es una alternativa frente al estrés oxidativo (Agarwal et al, 2013). El objetivo principal de esta Tesis fue evaluar el efecto antioxidante del GSH aplicado al medio de descongelación, sobre muestras seminales procedentes de tres especies de mamíferos con importancia en ganadería (porcina, bovina y caprina). En el primer trabajo (Experiencia 1) se evaluó el efecto de la adición de diferentes concentraciones de GSH (1 mM y 5 mM) al medio de descongelación de semen porcino. Las técnicas aplicadas incluyeron el análisis computarizado de imagen (CASA), citometría de flujo y ensayos de fecundación in vitro. En los resultados se observó una reducción en el número de espermatozoides viables capacitados, el número de espermatozoides con cambios en los grupos sulfhidrilo de sus proteínas de membrana, la producción de ERO y la condensación de la cromatina. Se obtuvo un incremento en el número de ovocitos penetrados y en la proporción de cabezas espermáticas descondensadas. Sin embargo, no se evidenció cambio alguno en los parámetros de movilidad evaluados con CASA. En el segundo trabajo (Experiencia 2), desarrollado en bovino, se realizaron ensayos similares a los de la experiencia 1 y se incluyeron nuevos análisis centrados en el estudio del ADN espermático, mediante las técnicas de TUNEL y SCSA. Además, se evaluaron los resultados de las técnicas de fecundación in vitro y desarrollo embrionario.. La adición de GSH supuso un incremento en el número de espermatozoides viables no capacitados, en el porcentaje de penetración y desarrollo embrionario in vitro. Se obtuvo un descenso en la producción de ERO y la fragmentación del ADN. No se observó mejoría en los parámetros de movilidad (CASA) y de reacción acrosómica. El último trabajo (Experiencia 3) tuvo lugar en caprino y fue diseñado con el objetivo de saber si la presencia de la forma reducida (GSH) y/o oxidadada del glutatión (GSSG) en el medio de descongelación tenía algún efecto sobre la funcionalidad espermática. Los ensayos incluyeron la valoración por CASA y citometría de flujo. La adición de GSH supuso un incremento en la movilidad y viabilidad espermática, el número de espermatozoides viables no capacitados y con membrana acrosomal intacta. Se observó una reducción en la producción de ERO y la condensación de la cromatina. A diferencia de lo ocurrido con GSH, la adición de GSSG no reveló efecto alguno sobre los parámetros seminales evaluados Estos trabajos demuestran que la adición de GSH al medio de descongelación seminal mejora los parámetros seminales y la fertilidad de las muestras. / SUMMARY Sperm cryopreservation is widely used in assisted reproduction procedures in humans and in animals, such us domestic livestock and endangered species. Although being the most effective method for the maintenance of seminal samples on a long term, cryopreservation has negative effect on semen quality and fertility (Aitken et al., 2004), because of the temperature drops and osmotic changes (Polge & Jakobsen, 1959). In addition to changes in the plasma membrane and in DNA of spermatozoa. (Pursel & Johnson, 1975; Holt, 2000). The spermatozoa and other masculine reproductive cells have different defense systems against free radicals, including antioxidant enzymatic and non-enzymatic defenses (Sies, 1993). In some conditions, an excess on the production of free radicals, a diminished antioxidant capacity or both situations can lead to an imbalanced condition called “oxidative stress” (Sies & Cadenas, 1985). Supplementation of antioxidants in the freezing and thawing media is an alternative against oxidative stress (Agarwal et al, 2013) In this sense, glutathione is one of the most ubiquitous and important antioxidants in the biological systems. (Gadea et al., 2005). The main aid of this thesis was to evaluate the antioxidant effect of GSH when added to the thawing solution in seminal samples from three species with importance in livestock. In the first study (Experiment 1) it was evaluated the addition of 3 different concentrations of GSH (1 mM and 5 mM) to the thawing solution of board semen, with an incubation time of 0 and 30 minutes. The techniques used were computer assisted sperm analysis CASA, flow cytometry and in vitro assays. The results obtained support the hypothesis, the presence of GSH had a protective effect in some of the seminal parameters studied. It was observed a reduction in the number of capacitated viable spermatozoa, in the number of sperm with changes in sulfhydryl group content in membrane proteins, in the ERO production as well as the chromatin condensation. An increase on the number of penetrated oocytes and decondensed sperm heads was also noted. Nevertheless, there was no evidence of any change in the motility scores recorded by CASA. The second study (Experiment 2), conducted on bovine, incudes apart from the studies developed on experiment 1 new analysis on sperm DNA, by TUNEL and SCSA techniques. Furthermore, it was evaluated the results of the in vitro techniques and the embryo development in presence of GSH in the thawing solution. The collected data from Experiment 2 showed concordance with the results from Experiment 1. With the addition GSH in the thawing solution, it was observed an increase in non-capacitated viable spermatozoa, in the percentage of penetration and in vitro development. ERO production and DNA fragmentation values were reduced. No changes were observed on sperm motility (CASA) o presence of spontaneous acrosome reaction. The third study (Experiment 3) on caprine, was designed with the propose of knowing if the presence of GSH 1 mM and 5 mM or GSSG 2.5 mM in the thawing media has any effect on sperm motility and sperm functionality. This assay included CASA evaluation and flow cytometry. In this case, the samples were incubated 30 and 60 minutes. The collected results demonstrated and increase on motility and sperm viability, in the number of non- capacitated viable spermatozoa and spermatozoa with acrosome membrane untouched. A reduction on the ERO production and on the condensation of chromatin was also observed. Opposite to GSH, the addition of GSSG did not show any effect on the evaluated seminal parameters independent to the exposure time. These studies demonstrate that the addition of GSH to the thawing media improves seminal parameters and fertility on the samples studied.

Con el presente trabajo hemos pretendido buscar los siguientes objetivos: La presencia de Wohlfahria magnifica in la provincia de Albacete, en relación con el padecimiento de las ovejas con W. magnifica en función de su aptitud productiva, tamaño del rebaño, nivel estructural e higiénico-sanitario y el manejo de los operarios. • Pare ello hemos utilizado la siguiente metodología. Se realizaron visitas-encuestas a 122 explotaciones. Se recogieron datos e información y ejemplares de larvas y moscas de W. magnifica. Seguidamente, se hizo el análisis estadístico se realizó con del programa R-gui para el cálculo del chi-cuadrado, con corrección de Yates, La Correlación de Pearsons y el Odds Ratio. • Los resultados obtenidos fueron los siguientes: De las 1.617 explotaciones existentes en el momento, se visitaron 122 ganaderías, lo que suponía el 7,54% de la muestra, repartida por 88 localidades. De un censo total de 646.893 ovejas se muestrearon 73.683 animales, únicamente de la especie ovina, lo que representaba el 11,39%. Del total de 122 explotaciones encuestadas, 117 se encontraban afectadas (95,90%). Del total de animales inspeccionados (73.683), 5.284 presentaban miasis cutáneas, de los cuales 5.283 estaban producidas por Wohlfahrtia magnifica, el 99,98% y 1 por Lucilia sericata, 0,02%. La prevalencia media provincial de animales afectados fue del 7,17%. La prevalencia encontrada, para las ovejas de aptitud láctea fue del 7,34%, frente a las ovejas de carne con el 7,03%. Esta prevalencia se situaba en el 7,16% para explotaciones con régimen semi-extensivo y del 7,29% para explotaciones extensivas. Asimismo, según la distribución geográfica, encontramos la prevalencia más elevada en la zona II con el 7,93%, frente a la prevalencia de la zona I, con el 7,33% y la zona III con el 6,52%. Un dato muy destacable de esta investigación, es el referente a que en la zona I, las ovejas de régimen de explotación semi-extensivo superaban en prevalencia 7,99% a las de régimen extensivo con el 6,88%. Este dato se hizo más relevante al saber que el 100% de las explotaciones semi-extensivas eran de producción láctea y que el 100% de las extensivas era de producción cárnica. En función del tamaño del rebaño, observamos que las de tamaños intermedio, entre 251 y 500 animales, tenían el mayor porcentaje con el 8,31%, seguida de las explotaciones pequeñas, menores de 250 animales con el 8,18%, y por último las mayores de 501 animales con el 6,74%. Según el nivel estructural de la explotación son las de peor nivel las que presentaban mayor prevalencia 9,98%, las de nivel regular y bueno 7,20% y 6,52%, respectivamente. De la misma manera que las explotaciones con peor nivel higiénico-sanitario, eran las de mayor prevalencia con el 10,53%, mientras que las de nivel regular y bueno representaban el 7,65% y 6,20% respectivamente. Y las explotaciones con peor manejo eran también la de mayor prevalencia con el 8,09%, seguida de las de manejo regular y bueno con el 7,76% y el 5,72%. Los machos tienen una mayor predisposición 57,14% al padecimiento de una wohlfahrtiosis, 22 veces superior a las hembras 5,71%. Las zonas más afectadas en ovino eran las zonas genitales 80,47%, seguida de las podales con el 5,24% y las auriculares con el 3,27%. Si estos porcentajes se desglosaban por sexos se aprecia que en machos, la afección en el pene se elevaba al 94,46%, en la cabeza el 3,27% y las afecciones podales y oculares con el 0,59%, mientras que en hembras entre la zona vulvar y perianal la prevalencia fue del 76,39%, la zona podal el 6,60%, la zona auricular al 4,15% y la mamaria al 4,11%. Por otro lado, las ovejas de pura raza segureña de aptitud cárnica y régimen extensivo con el 7,74% son más propensas que las ovejas de p-r manchega de la misma aptitud y régimen de explotación 7,12%. En 16 de las 122 explotaciones se pudo observar animales afectados por wohlfahrtiosis sin que estos hubieran salido de la explotación, dato relevante, pues nos demuestra que Wohlfahrtia magnifica es capaz de producir miasis en animales en régimen de explotación intensivo. / With this work we have tried to look for the following objectives: The presence of Wohlfahrtia magnifica in the Albacete province and the interrelationship of the suffering of sheep with W. magnifica according to productive capacity, folck size, structural and hygienic-sanitary level and livestock handling systems. • To do this we used the following methodology: Visits-sampled survey of 122 farms. With the collection of information data and samples of larvae and Spotted Flesh Fly. Subsequently, the statistical analysis was performed with the R-Gui program for the calculation of chi-square, with Yates correction, Pearsons Correlation and Odds Ratio. • The results were as follows. Of the 1,617 farms currently in existence, 122 livestock farms were visited, representing 7.54% of the sample, spread over 88 localities. From a total census of 646,893 ovine livestock, 73,683 animals were sampled, all of which were sheep and which represented 11.39% of the total. A total of 122 farms were surveyed, and 117 (95.90%) of those were found to be affected. Of the 73,683 animals inspected, 5,284 presented symptoms of skin myiasis, 5,283 (99.98%) of which were caused by W. magnifica and one (0.02%) by Lucilia sericata. The average provincial prevalence of infected animals was 7.17%. The prevalence in dairy sheep was 7.34% compared to 7.03% in sheep bred for meat. This prevalence was situated at 7.16% for farms with a semi-extensive pastoral system and 7.29% for extensive farms. Likewise, according to geographical distribution, the highest prevalence was found in zone II with 7.93% as opposed to the prevalence in zone I, of 7.33% and in zone III of 6.52%. A key statistic, revealed by this study, refers to zone I and the fact that the sheep belonging to the semi-extensive farms of that area exceeded a prevalence of 7.99% in comparison with that of 6.88% of those of the extensive farm systems. This information acquires more significance when it is considered that 100% of the semi-extensive farms were dedicated to milk production and 100% of the extensive farms were meat producing. According to the flock size, it can be observed that the intermediate sizes of between 251 and 500 animals had the highest percentage of 8.31% followed by the small farms, with less than 250 animals, with 8.18%, and finally the largest ones with flocks of over 501 animals, with 6.74%. According to the structural level of the farm, it is those with the poorest level that presented the greatest prevalence 9.98%. The prevalence of those with a normal or good level was 7.2% and 6.25%, respectively. Similarly, the farms with the lowest sanitary hygiene levels were those with the greatest prevalence at 10.53%, while those with a normal and good level represented 7.65% and 6.20%, respectively. Furthermore, the farms with the worst livestock handling systems were also those with the greatest prevalence at 8.09%, followed by those with normal handling systems and good handling systems at 7.76% and 5.72%. Males are more predisposed 57.14% to suffering wohlfahrtiosis, 22 times greater than the females 5.71%. The most affected areas for sheep were the genital areas at 80.47%, followed by the feet at 5.24% and the ears at 3.27%. If these percentages are broken down according to sex, it can be observed that in males, the infection on the penis reached 94.46%, that of the head 3.27% and that of the feet and eyes 0.59%. With regards to females, prevalence between the vulva and perianal areas reached 76.39% that of the area of the foot was 6.60%, the area of the ears 4.15% and the mammary area 4.11%. However, it was concluded that the purebred Segureña sheep, bred for meat and with an extensive pastoral system, at 7.74% were more prone to the infection than purebred Manchega sheep raised for meat and with the same extensive system at 7.12%. On 16 of the 122 farms, it was possible to observe animals affected by wohlfahrtiosis without having left the farm. This is important as it shows that W. magnifica is capable of producing myiasis in animals in an intensive farming system.

Orientador: Renée Laufer Amorim / Banca: Noeme Sousa Rocha / Banca: Antonio Carlos Alessi / Resumo: Os mastócitos são células provenientes do precursor CD34 da medula óssea, o mastocitoma (MCT) pertence ao grupo das neoplasias de células redondas e possui etiologia desconhecida, é a neoplasia cutânea mais comum em cães, sem predileção por raça, sexo ou idade. Seu comportamento biológico é bastante agressivo e de alta frequência. A graduação histopatológia proposta Patnaik et al., (1984), é o critério mais utilizado para a graduação histopatológica da neoplasia sendo o grau 1 uma neoplasia bem diferenciada, o grau 2 moderadamente diferenciada e o grau 3 pouco diferenciada ou anaplásica. O presente trabalho teve como objetivos a avaliação do ínidice proliferativo e apoptotico dos MCT em lâmina de Microarranjo de tecido (TMA) utilizando a técnica de imunoistoquiímica e TUNEL. Para tal utilizou-se 166 casos de MCT compondo um arranjo em matriz de amostra tecidual, ou tissue microarray (TMA) com 190 amostras, sendo avaliado o padrão de marcação da proteína Kit, a proliferação celular com o anticorpo primário Ki67 e apoptose com o anticorpo primário caspase-3 clivada e pela técnica da Terminal Deoxynucleotidyl Transferase Mediated dUTP Nick end Labeling Assey (TUNEL). Quando os casos foram avaliados quanto à imunoexpressão de c-KIT, os tumores (únicos e múltiplos) com expressão citoplasmática da proteína KIT tiveram uma taxa proliferativa maior e uma menor taxa apoptótica quando comparados aos com expressão membranosa. Os MCTs grau 3 tiveram um maior índice proliferativo (imunoexpressão de Ki67) quando comparados aos tumores de graus 1 e 2. Não foi observada correlação entre a imunomarcação do anticorpo primário caspase-3 clivada e a técnica de TUNEL, sendo que a técnica de TUNEL se mostrou mais eficaz na avaliação da apoptose que a imunomarcação para caspase-3 clivada / Abstract: Mast cells are from precursor CD34 cells bone marrow, the mast cell tumor (MCT) belongs to the group of round cell malignancies and has unknown etiology, is the most common skin cancer in dogs without distinction of race, sex or age. Its biological behavior is very aggressive and high frequency. The histopathological grading proposal from Patnaik et al., (1984), is the most commonly used criterion for the histopathological grade of the tumor is a tumor grade 1 are well-differentiated, grade 2 are moderately differentiated and grade 3 are poorly differentiated or anaplastic. This study aimed to evaluate the proliferative and apoptotic index of MCT in blade tissue microarray (TMA) using the TUNEL technique and immunohistochemistry. To this end we used 166 cases of MCT composing an tissue microarray (TMA) with 190 samples, and evaluated the pattern of protein labeling kit, cell proliferation, with the primary antibody Ki67 and apoptosis with primary antibody and cleaved caspase-3 by the technique of terminal deoxynucleotidyl transferase dUTP Nick End Labeling Mediated Assey (TUNEL). When the cases were evaluated for immunoexpression of c-KIT, tumors (single and multiple) with cytoplasmic KIT protein expression had a higher proliferative rate and a lower apoptotic rate when compared to those with membranous expression. Grade 3 MCTs had a greater proliferative index (Ki67 immunostaining) when compared with tumor grades 1 and 2. No correlation was observed between the immunostaining of primary antibody caspase-3 cleavage and TUNEL technique, and TUNEL technique was more effective in assessing apoptosis that immunostaining for cleaved caspase-3 / Mestre

Orientador: Maria Cecília Rui Luvizotto / Resumo: O objetivo desta pesquisa foi avaliar o comportamento dos biomarcadores renais dimetilarginina simétrica (SDMA), creatinina sérica (CRs) e a relação proteína:creatinina urinária (UPC) em gatos aparentemente saudáveis (n: 20) e gatos portadores de doença renal crônica (n:10). No grupo de gatos com doença renal crônica (DRC) os valores médios de SDMA, CRs e UPC foram 20,4 ± 3,1 µg/dL, 2,4 ± 0,7 mg/dl e 0,6 ± 0,23, respectivamente. Para o grupo aparentemente saudável a SDMA, CRs e UPC obtidos foram 9,8 ± 1,8 µg/dL, 0,9 ± 0,4 mg/dl e 0,28 ± 0,08. Para o grupo positivo para DRC, o coeficiente de correlação entre a SDMA e CRs foi r = 0,52, indicando ser correlação moderada entre estes marcadores renais. Pelo exposto conclui-se que os gatos sintomáticos para DRC necessitam serem subestadiados pelos critérios da IRIS (2017) (UPC, CRs, PAS e SDMA) e execução de medidas de renoproteção como tentativa de controle da progressão da DRC presente. Para os gatos aparentemente saudáveis o comportamento da SDMA deu-se dentro da normalidade para a espécie, com um valor de UPC limítrofe, sugerindo monitoramento e acompanhamento trimestral ou semestral, conforme avaliação clínica. / Abstract: The objective of this study was to evaluate the behavior of the renal biomarkers dimethylarginine symmetric (SDMA), serum creatinine (CRs) and protein: urinary creatinine ratio (UPC) in apparently healthy cats (n: 20) and cats with chronic kidney disease (10). In the group of cats with chronic kidney disease (CKD) the mean values of SDMA, CRs and UPC were 20,4 ± 3,1μg / dL, 2,4 ± 0,7 mg / dl and 0,6 ± 0,23, respectively. In the apparently healthy group the SDMA, CRs and UPC obtained were 9,8 ± 1,8 μg / dL, 0,9 ± 0,4 mg/dl and 0,28 ± 0,08. In the group of cats with chronic kidney disease the correlation of SDMA and CRs was r = 0.52, indicating a moderate correlation between these renal biomarkers. Based on those findings concluded that cats with CKD require a detailed diagnosis by IRIS (2017) (UPC, CRs, PAS and SDMA) to perform kidney protection and attempt to control the progression of CKD. For apparently healthy cats, the behavior of SDMA was the normal range for the species, with a borderline UPC value, suggesting monitoring and quarterly or semi-annual follow-up, according to clinical evaluation. / Doutor

El proyecto consiste en el diseño, por un lado, de una Clínica Veterinaria de Animales Menores de jerarquía interdistrital que se complementa con un Albergue de Animales (perros y gatos). El objetivo de este último es poder brindar a los animales rescatados un espacio adecuado a sus necesidades durante el tiempo que pasen en este refugio. Asimismo, la Clínica atiende tanto a usuarios externos como a los usuarios del Albergue. Por otro lado, se tiene una zona de capacitación y difusión dirigida a veterinarios, estudiantes de veterinaria y personas interesadas en temas relacionados al cuidado y tenencia responsable de mascotas. El terreno del proyecto se ubica en el distrito de Santiago de Surco. Para esta elección se tuvo en consideración los siguientes requerimientos: Zona céntrica para los demás albergues existentes; lugar en el que las personas del entorno puedan permitirse pagar tratamientos a sus mascotas y cercanía al “Colegio Médico Veterinario del Perú”. En cuanto al diseño arquitectónico, se optó por separarlo en dos volumetrías, esto debido a que se quiere evitar que los usuarios de las dos grandes áreas de este proyecto se mezclen. Adicionalmente, se busca controlar el ruido hecho por los animales, sin que afecte a las personas que se encuentran en la zona de capacitación. Por último, estos volúmenes están ubicados y diseñados de modo que se genera una relación visual entre ambos. / The project consists of the design, on the one hand, of a Veterinary Clinic of Minor Animals of interdistrict hierarchy that is complemented by an Animal Shelter (dogs and cats). The objective of the latter is to be able to provide the rescued animals with an adequate space for their needs during the time they spend in this refuge. Also, the Clinic serves both external users and users of the Shelter. On the other hand, there is a training and dissemination area for veterinarians, veterinary students and people interested in issues related to the care and responsible tenure of pets. The project land is located in the district of Santiago de Surco. For this election, the following requirements were taken into consideration: Central area for other existing hostels, place where the people of the environment can afford to pay treatments to their pets and Closeness to the "Colegio Médico Veterinario del Perú". For the architectural design, we chose to separate it into two volumes, this is because we want to avoid that the users of the two large areas of this project mix. Additionally, it seeks to control the noise made by the animals, without affecting the people who are in the training area. Finally, these volumes are located and designed so that a visual relationship between them is generated. / Tesis

Procura aprender o significado psicologico possuido pelo animal de estimacao no cotidiano das pessoas, atraves do exame de relatos de donos e nao-donos de animais. Realiza entrevistas com adultos de ambos os sexos, dos quais 13 convivem com animais de estimacao, 6 nao possuem nem pretendem a posse e 3 pretendem adquirir um animal no futuro. Inclui, tambem, no estudo, dados oriundos de observacoes da experimentadora em sua clinica veterinaria e de relatos espontaneos ouvidos em situacoes diversas. Apos a gravacao e transcricao literal das entrevistas seleciona e analisa as modalidades e razoes de obtencao de um animal; cuidados dispensados no seu tratamento e sobrevivencia; as maneiras como se efetua a comunicacao entre o dono e seu animal; as formas de rompimento do vinculo entre dono e animal de estimacao e as funcoes que o animal exerce junto ao ser humano.Verifica que a presenca de um animal de estimacao pode trazer beneficios psico-sociais ao dono, derivados nao do animal no sentido biologico, mas de um animal imaginario e que desempenha funcoes e papel de acordo com as necessidades de cada ser. Enfatiza a importancia do conhecimento dos aspectos psicologicos do relacionamento ser hunamo-animal para o profissional psicologo e para o veterinário / The experience related by a number of persons regarding their relationship with pet animals formed the background material from which the attempt was made to extract the psychological significance of such pets in everyday life. The underlying data were obtained from interviews with 13 adults who owned pets and 9 adult non-pet-owners, 3 of which intended to acquire a pet animal. All interwiews were tape-recorded and subsequently transcribed verbatim. Further data - when deemed necessary for completeness and back-up purposes - were obtained from veterinary medical records. After pinpointing and eliminating as irrelevancies certain fortuitious nos-recurring elements, conclusions could clearly be drawn on the following: motivation for acquiring a pet; forms of such an acquisition; routines and patterns of caring for, and ways to communicate with the animal; vicissitudes for both owner and pet; and finally, options to continue, discountinue or temporarily interrupt the relationship

A Fazenda Três Lagoas, localizada no município de Angatuba, região do Alto Paranapanema (centro-sul do Estado de São Paulo), é uma área na qual houve introdução de espécies exóticas, como Eucalyptus sp e Brachiaria sp, que fragmentaram o habitat de cerrado original. Uma vez em que foi estabelecida atividade humana em um ambiente antes natural, houve expansão do contato entre humanos e animais domésticos com as populações de animais silvestres; sendo a influência destes fatores sobre o ecossistema local ainda pouco conhecido. O presente estudo teve como objetivo pesquisar a exposição de pequenos mamíferos silvestres e animais domésticos da região a agentes infecciosos e transmissores de zoonoses (Leptospira spp., Lyssavirus, Toxoplasma gondii, Neospora caninum e Rickettsia spp.). O levantamento dos patógenos nos animais silvestres revelou resultados sorológicos positivos para Leptospira spp. (5,5%, n= 73), Rickettsia spp. (7,9%, n=38) e Toxoplasma gondii (16,1%, n= 56). Em relação aos animais domésticos, foram encontrados 40% de cães (n=10) soropositivos para N. caninum, 20% de soropositivos para Leptospira spp. e T. gondii (n=10), e 23,27% dos cães (n=11) apresentaram títulos de anticorpos menores ou iguais a 0,5 UI/ml para Raiva. Os bovinos (n=42) apresentaram resultados sorológicos positivos para N. caninum (23,8%) e Leptospira spp. (40,5%). 47,6% do gado não havia sido vacinado ou a vacinação para o vírus rábico era desconhecida. Dos 22 bovinos conhecidamente vacinados para a Raiva, 36% foram pouco reagentes ao exame sorológico. Os resultados apresentados, além de contribuírem para a avaliação do status sanitário local, contribuem para o maior conhecimento da interface entre fauna doméstica e silvestre em agroecossistemas. / Três lagoas farm, which is located in Angatuba municipality, in High Paranapanema (southeast of São Paulo state) is an area where exotic species were introduced, such as Eucalyptus sp and Brachiaria sp, which fragmented the habitat in the original Cerradão (a tropical savanna ecoregion). Once the human activity has been established in a natural environment, the contact among human beings and domestic animals with wild animal populations has increased. Nevertheless, the influence of such factors on the ecosystem has not been clarified yet. The aim of the present study was to research the exposition between wild small mammals and domestic animals of this area against infecctious and transmitters of zoonoses agents (Leptospira spp., Toxoplasma gondii, Lyssavirus, Neospora caninum and Rickettsia spp.). The raising of pathogens in the wild animals revealed positive serological results for Leptospira spp. (5,5%, n= 73), Rickettsia spp. (7,9%, n=38) e Toxoplasma gondii (16,1%, n= 56). In relation to the domestic animals, were showed 40% of dogs (n=10) seropositives for N. caninum, 20% were seropositives for Leptospira spp. and T. gondii, and 23.7% of dogs presented antibody titers less than or equal to 0,5 UI/ml for Rabies. The cattle (n=42) had serologic results positives for N. caninum (23,8%) and for Leptospira spp. (40,5%). 47.6% of the cattle had not been vaccinated or vaccination for the rabies virus was unknown. Of the 22 cattle known to be vaccinated for rabies, 36% were less reactive to serologic test. These results presented, besides evaluating the local sanitary status, they can contribute to the greater understanding of the interface between domestic and wild fauna in agricultural ecosystems.