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Situación Epidemiológica Y Clínica De La Piroplasmosis Equina En Áreas Endémicas Como La Península Itálica E Ibérica

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Situación Epidemiológica Y Clínica De La Piroplasmosis Equina En Áreas Endémicas Como La Península Itálica E Ibérica UNIVERSIDAD COMPLUTENSE DE MADRID FACULTAD DE VETERINARIA Departamento de Sanidad Animal TESIS DOCTORAL Situación epidemiológica y clínica de la piroplasmosis equina en áreas endémicas como la Península Itálica e Ibérica Epidemiological and clinical situation of equine piroplasmosis in endemic areas such as Italian and Iberian peninsulas / MEMORIA PARA OPTAR AL GRADO DE DOCTOR PRESENTADA POR Leticia Elisa Bartolomé del Pino Directores Aránzazu Meana Mañes Gian Luca Autorino Madrid, 2017 © Leticia Elisa Bartolomé del Pino, 2017 UNIVERSIDAD COMPLUTENSE DE MADRID FACULTAD DE VETERINARIA DEPARTAMENTO DE SANIDAD ANIMAL SITUACIÓN EPIDEMIOLÓGICA Y CLÍNICA DE LA PIROPLASMOSIS EQUINA EN ÁREAS ENDÉMICAS COMO LAS PENÍNSULAS ITÁLICA E IBÉRICA EPIDEMIOLOGICAL AND CLINICAL SITUATION OF EQUINE PIROPLASMOSIS IN ENDEMIC AREAS SUCH AS ITALIAN AND IBERIAN PENINSULAS TESIS DOCTORAL LETICIA ELISA BARTOLOMÉ DEL PINO MADRID, 2017 1 UNIVERSIDAD COMPLUTENSE DE MADRID FACULTAD DE VETERINARIA DEPARTAMENTO DE SANIDAD ANIMAL SITUACIÓN EPIDEMIOLÓGICA Y CLÍNICA DE LA PIROPLASMOSIS EQUINA EN ÁREAS ENDÉMICAS COMO LAS PENÍNSULAS ITÁLICA E IBÉRICA EPIDEMIOLOGICAL AND CLINICAL SITUATION OF EQUINE PIROPLASMOSIS IN ENDEMIC AREAS SUCH AS ITALIAN AND IBERIAN PENINSULAS MEMORIA PARA OPTAR AL GRADO DE DOCTOR PRESENTADA POR LETICIA ELISA BARTOLOMÉ DEL PINO DIRECTORES ARÁNZAZU MEANA MAÑES GIAN LUCA AUTORINO MADRID, 2017 Leticia E. Bartolomé del Pino, 2017 3 UNIVERSIDAD COMPLUTENSE DE MADRID FACULTAD DE VETERINARIA DEPARTAMENTO DE SANIDAD ANIMAL SITUACIÓN EPIDEMIOLÓGICA Y CLÍNICA DE LA PIROPLASMOSIS EQUINA EN ÁREAS ENDÉMICAS COMO LAS PENÍNSULAS ITÁLICA E IBÉRICA EPIDEMIOLOGICAL AND CLINICAL SITUATION OF EQUINE PIROPLASMOSIS IN ENDEMIC AREAS SUCH AS ITALIAN AND IBERIAN PENINSULAS TESIS DOCTORAL LETICIA ELISA BARTOLOMÉ DEL PINO MADRID, 2017 5 El trabajo presentado en esta tesis se ha realizado bajo la dirección de Gian Luca Autorino y Aránzazu Meana Mañes en el Centro de Referencia Nacional de las Enfermedades de los Équidos del Istituto Zooprofilattico Sperimentale delle Regioni Lazio e Toscana (IZSLT) en Roma, Italia con fondos del proyecto IZSLT 16/11 del Ministerio de Sanidad Italiano y en el Departamento de Sanidad Animal de la Facultad de Veterinaria de la Universidad Complutense de Madrid. Durante la realización del mismo la autora ha disfrutado de una beca Erasmus Prácticas concedida por la Universidad Complutense y una beca de apoyo a la investigación en cotutela ofrecida por el Ministerio de Asuntos Exteriores Italiano. Asimismo, se han realizado estancias en la Universidad Federico II de Nápoles (Italia). 7 Centro di Referenza Nazionale per le Malattie degli Equini, Istituto Zooprofilattico Sperimentale delle Regioni Lazio e Toscana - Roma (Italia) y Departamento de Sanidad Animal de la Facultad de Veterinaria de la Universidad Complutense de Madrid. Gian Luca Autorino, director del Centro di Referenza Nazionale per le Malattie degli Equini (CERME) dell’Istituto Zooprofilattico Sperimentale delle Regioni Lazio e Toscana - Roma (Italia) y M. Aránzazu Meana Mañes profesora titular del Departamento de Sanidad Animal de la Facultad de Veterinaria de la Universidad Complutense de Madrid. CERTIFICAN: Que la tesis doctoral «Situación epidemiológica y clínica de la piroplasmosis equina en áreas endémicas como las Penínsulas Itálica e Ibérica.» ha sido realizada por la licenciada en Veterinaria Dña. Leticia Elisa Bartolomé del Pino en ambos centros bajo nuestra dirección y estimamos que reúne los requisitos exigidos para optar al Título de Doctor por la Universidad Complutense de Madrid. Gian Luca Autorino M. Aránzazu Meana Mañes 9 Dedicada a mi familia y a quien se siente parte de ella 11 “C'est le temps que tu a perdu pour ta rose qui fait ta rose si importante.” A. de Saint-Exupéry 13 AGRADECIMIENTOS En primer lugar quiero agradecer a mis tutores Gian Luca Autorino y Aránzazu Meana la ayuda prestada, los consejos e ideas que me han ayudado a aprender a mirar la ciencia desde otro prisma. Agradezco también a los técnicos y veterinarios del CERME del IZSLT de Roma el trato recibido durante este tiempo, a Teresa, Francesca, Roberto, Raffaele e Ida gracias por su disponibilidad, ayuda y paciencia, habéis sido grandes maestros. Gracias también a todo el reparto de Biotecnologie del IZSLT en especial a Antonella Cersini que me ha hecho descubrir un nuevo mundo llamado Biología Molecular muy abstracto y a veces desesperante pero siempre apasionante, a los técnicos Valeria, Silvia, Raffaella y Maurizio, gracias por la ayuda, los trucos y las masterclass de secuencias. Otro agradecimiento al profesor Vincenzo Veneziano y a los demás componentes del grupo de investigación de la Facultad de Veterinaria de la Universidad Federico II de Nápoles por involucrarme en su estudio y la gran acogida que recibí, recuerdo con mucho cariño esa estancia. A los propietarios de los animales por su gran disponibilidad y a Mari Cruz Llorente por su inestimable ayuda en los muestreos. A Ricardo García, de la UCM por la ayuda prestada con la Estadistica. A la gente de la Facultad: las chicas del 126 Ana, Rocio y Valentina, que siempre me han echado una mano, a Julia y Miguel, que también me han ayudado entre llaves, horarios intempestivos y documentos. A Marta Hernández, María Alcaide y Daniel Beltrán por la ayuda prestada con las traducciones y las correcciones, y quien que me ha ayudado a resolver imprevistos logísticos, Isaac de Bedoya y Patri Alba. A mis amigos y residentes en el mundo, todas las experiencias vividas aquí y allí sé que nos unirán estemos donde estemos. Gracias por vuestro apoyo. Ai regà de Biotecnologie mediche que cambiaron positivamente mi concepto de italiano medio. A mi familia, que siempre me ha apoyado en la distancia durante mis andanzas, mis locuras y mis luchas contra molinos de viento. A Claudio, bambino speciale, grazie di tutto sempre, no hay “rigatoni” suficientes para agradecerte lo que haces por mí. A mi Layka, la pequeña doctora honoris causa, casi la única constante en este tiempo, mi compañía diurna y nocturana en licenciatura, master y doctorado. Un gracias final a todo aquel que me animó cuando lo necesitaba durante este recorrido, y también una disculpa a todas las personas a las que no he podido dedicar el tiempo que se merecen en muchos momentos. 15 INDEX INTRODUCTION AND OBJECTIVES ........................................................................................................ 7 EQUINE PIROPLASMOSIS: STATE OF ART…. ........................................................................................ 30 CHAPTER 1: DIAGNOSTIC TECHNIQUES ............................................................................................. 55 Study 1: Comparison of blood smear, serological and molecular techniques for the diagnosis of equine piroplasmosis Abstract…. ................................................................................................................................. 61 Introduction… ........................................................................................................................... 61 Materials and methods ............................................................................................................. 62 Results and discussion… .......................................................................................................... .66 Conclusions ............................................................................................................................... 70 References… .............................................................................................................................. 71 CHAPTER 2: EPIDEMIOLOGY AND PHYLOGENETICS .......................................................................... .75 Study 2.1: Serological detection of Theileria equi and Babesia caballi infections in horses and donkeys in areas of Central Spain and risk factors associated. Abstract ..................................................................................................................................... 85 Introduction… ........................................................................................................................... 85 Materials and methods. ........................................................................................................... .86 Results ....................................................................................................................................... 88 Discussion ................................................................................................................................. 89 Conclusions ............................................................................................................................... 93 References….. ............................................................................................................................ 93 Study 2.2: Molecular and serological detection of Theileria equi and Babesia caballi infections in Equids in areas of Central-Southern Italy. Abstract... .................................................................................................................................. 99 Introduction….… ..................................................................................................................... 100 Materials and methods ........................................................................................................... 101 Results ....................................................................................................................................
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