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Tesis Doctoral UNIVERSIDAD COMPLUTENSE DE MADRID FACULTAD DE VETERINARIA Departamento de Sanidad Animal TESIS DOCTORAL CARACTERIZACIÓN FENOTÍPICA Y GENÉTICA DE AISLADOS DE Pasteurella multocida OBTENIDOS DE GANADO PORCINO MEMORIA PARA OPTAR AL GRADO DE DOCTOR PRESENTADA POR Nerea García Benzaquén Bajo la dirección de los doctores: José Francisco Fernández-Garayzábal Fernández, Joaquín Goyache Goñi y Ana Isabel Vela Alonso Madrid, 2010 ISBN: 978-84-693-2407-3 UNIVERSIDAD COMPLUTENSE DE MADRID FACULTAD DE VETERINARIA DEPARTAMENTO DE SANIDAD ANIMAL TESIS DOCTORAL CARACTERIZACIÓN FENOTÍPICA Y GENÉTICA DE AISLADOS DE Pasteurella multocida OBTENIDOS DE GANADO PORCINO Dña. Nerea García Benzaquén 2009 D. José Francisco Fernández-Garayzábal Fernández, D. Joaquín Goyache Goñi y Dña. Ana Isabel Vela Alonso, Profesores titulares del Departamento de Sanidad Animal de la Facultad de Veterinaria CERTIFICAN: Que la tesis doctoral que lleva por título “Caracterización fenotípica y genética de aislados de Pasteurella multocidda obtenidos de ganado porcino” ha sido realizada por la licenciada en Veterinaria Dña. Nerea García Benzaquén en el Departamento de Sanidad Animal de la Facultad de Veterinaria de la Universidad Complutense bajo nuestra dirección, y estimamos que reúne los requisitos exigidos para optar al título de Doctor por la Universidad Complutense. Y para que así conste, a efectos legales, firman y sellan el presente certificado en Madrid, a 14 de mayo de 2009. Fdo. J. F. Fernández-Garayzábal Fdo. A. I. Vela Fdo. J. Goyache Queda prohibido llorar sin aprender, levantarte un día sin saber qué hacer, tener miedo a tus recuerdos. Queda prohibido no sonreír a los problemas, no luchar por lo que quieres, abandonarlo todo por miedo, no convertir en realidad tus sueños. Pablo Neruda Todo aquél que piense que la vida es desigual tiene que saber que no es así Que la vida es una hermosura, hay que vivirla Todo aquel que piense que está solo y que está mal Tiene que saber que no es así Que en la vida no hay nadie solo, siempre hay alguien Ay, no hay que llorar Que la vida es un carnaval y es más bello vivir cantando...... Celia Cruz A Mis Padres AGRADECIMIENTOS Y por fin llegó el fin tan ansiado, después de tanto tiempo…¡viva! Cualquiera que me conozca lo más mínimo sabe que lo mío no es la capacidad de síntesis y que soy bastante despistada. A pesar de ello intentaré resumir y no olvidarme de nadie. Si no consigo alguna de las dos cosas pido mil disculpas. En primer lugar agradecer sinceramente a mis directores de tesis todo el trabajo y el apoyo que me han brindado durante todo este tiempo. Quisiera dar las gracias a José F. Fernández-Garayzábal por sus revisiones críticas y precisas, siempre con gran prontitud y siempre dispuesto a discutir mis ideas. A Ana Isabel Vela el tiempo y la dedicación que ha destinado a las infinitas correcciones. Y por último a Joaquín Goyache por su esfuerzo y por aportar sus valiosas opiniones e infundir estilo en la escritura de esta tesis. Además, querido Joaco, expresarte mi infinita gratitud por haber depositado tu confianza en mí hace mucho tiempo, sin apenas conocerme, y haberme dado la oportunidad que ha hecho que hoy haya llegado a este punto. Gracias, sinceramente, por animarme y apoyarme siempre, por escucharme, por consolarme en los momentos difíciles y por ser mi amigo. Gracias a Lucas Domínguez, por haber sacrificado sus ganas en este proyecto para el beneficio del mismo. Además por infundir su entusiasmo y su optimismo día a día, por ayudarme siempre a buscar la mejor solución y por hacerme sentir que puedo contar con él siempre que lo necesito (aunque su agenda lo complique a veces). Muchas gracias a toda la gente que ha puesto su granito de arena directamente en esta larga tesis. En primer lugar a Almudena, sin la cual hubiera sido imposible no volverme loca con el enrevesado mundo del campo pulsado. Gracias por tu paciencia y ayuda, Almu. Además a todo el grupo de veterinarios de campo sin los cuales hubiera sido imposible conseguir todas las muestras. Gracias a Gema (pequeña pero con gran dosis de valentía y arrojo) y a todo su equipo: Jose Cebolla (sufridor resignado de mi banda sonora en el despacho), Carmen, Fidel y Cristina. Y, por supuesto, gracias a todas las granjas, ganaderos y veterinarios que colaboraron y facilitaron el trabajo a este grupo. Gracias a todos los que me ayudaron en el laboratorio, porque habéis sido fundamentales para que este trabajo se haya conseguido y por haber logrado que fueran divertidos y amenos los días de mayor rutina. Gracias a mis primeras compañeras, Laura Delgado (quien me enseñó los trucos de laboratorio), Cris, Laura Cuesta y Pilar Vera. Tengo que dar las gracias a toda la gente que pasó por el grupo de bacteriología con quienes compartí infinitas horas de trabajo y amistad. Nuestra “jefa-madre” Pilar Liébana, siempre con la sonrisa a punto, dispuesta a echar una mano. Quique, gracias por tus momentos geniales, por sacarme una sonrisa hasta debajo del agua y gracias Delia por tu compañerismo y colaboración. También gracias a Ruth y a MariCarmen. Mención especial se merecen mis últimos y queridísimos compañeros de aquella etapa. Darles las gracias a ambos por mil cosas, por haber compartido horas de duro trabajo, risas, lágrimas, muchos bailes, confesiones, carcajadas, más y más bailes...Maguis, gracias porque las innumerables experiencias juntas (buenas y malas) nos han unido para siempre, y gracias cuidar de mi para que no perdiera el bolso y para que no metiera los dedos en el café. Jose Luis, gracias por entenderme a la perfección, por ser el tío más divertido del mundo mundial y porque casi consigues que aprenda a silbar. Me ha encantado conoceros y os echo muchísimo de menos. Gracias también a Mateo del Pozo y a Elena Vizcaíno por la aportación de material gráfico y al Dr. Rodríguez-Ferri por el material para las pruebas serológicas. A todos los compañeros de VISAVET, mi segunda casa, donde he conocido a tanta gente estupenda: Empezaré por Julito, imprescindible en estos años y espero que en los venideros. Gracias por tu ingenio, por hacerme reir sin descanso, por ayudarme tanto y confiar en mí, por tu brillantez y tesón en el trabajo, por nuestras incansables charlas…y por seguir cantando conmigo dúos imposibles. A Javichu, gracias por tu humor ácido pero cariñoso, porque sé que puedo contar contar contigo en cualquier momento y porque sin ti no estaría al día de lo que pasa por el mundillo rosa. Gracias a Soni, simplemente por ser más buena que el pan, por enseñarme tantas y tantas cosas con infinita paciencia. Gracias a Laura de las Heras, de las personas más eficientes que conozco, capaz de desenvolverse con soltura con el intrincado mundo del papeleo y también de conseguir que nos reunamos de vez en cuando todos. Y gracias a ambas por ayudarme a darle forma a esta tesis en los últimos momentos. Gracias a Sergio, por su inestimable amistad, por su ayuda con la informática y el diseño de mis trabajos, incluido este. A la “topilla” Marta, porque ha sido un placer contar contigo en el trabajo estos últimos años (a pesar de los sustos que me das de vez en cuando). Mil gracias por compartir las responsabilidades conmigo mano a mano. Por ser un ejemplo de perseverancia y constancia, por no negar nunca la ayuda a nadie y por contagiarnos tu entusiasmo. A Mazariegos y a Irene por simpáticas, por hacer del despacho un lugar francamente agradable para trabajar. A las tres muchas gracias porque en momentos de crisis vuestro “no te preocupes, te ayudamos” ha sido mi salvación. Gracias también a Lucía de Juan, jefa en un breve pero valioso tiempo en el que me sentí muy bien arropada. Gracias a toda la gente que ha compartido conmigo horas de café y de pasillo y que me han alegrado los días como Tania, Bea Tames, Ester, Lucía Palacios, Bea Romero, Sabrina, Elena, Elisa, Leydis, Vero, David, Pedro, Carlos, Susana, Isa, María García, Fanny, Tiru, Mauricio, sevemures varios, el resto del grupo de ZTA y el de micobacterias y un larguísimo etc. Como no, a Marga por ser tan maja y por no enfadarse nunca. También a Laura Torres, a Carmen y a Arancha. A todos los profesores, becarios, técnicos, etc. que han pasado por mi vida estos años que han colaborado conmigo y que me han sabido aconsejar como Miguel Angel Moreno, Alicia Aranaz, Jose Manuel Sánchez-Vizcaíno (y sus chicas), Alicia Gibello, Victor Briones, Bruno (y sus chicos), Concha Porrero, Ana Mateos, Mónica Suarez, Mar Blanco, etc. Agradecer, a pesar de la distancia, a la Dra. Rosario Esperanza Galván y a todo su equipo de la Universidad Autónoma de Méjico por haberme acogido como a una más y portarse tan bien conmigo. Ya fuera del ámbito profesional agradecer a todos mis amigos el haber compartido innumerables días buenos y que me han soportado y animado en los días malos. Gracias a Martita por mantener nuestra eterna amistad y porque sin ella no hubieran sido llevaderas las interminables horas de estudio en la carrera. A Pilar, que aún en la distancia es capaz de contagiarme su espíritu de lucha y superación, además de su cariño. Al resto de compañeros de carrera, especialmente a la gente de Vekavel: Sebas, Vane, Ainhoa, Asier, Chara, Jaco, Elías, Teclas, etc. A las que empezaron siendo “las amigas de mi hermana” y ahora son las mías por derecho propio y con las que he compartido tantos y tantos buenos momentos. Las “supernenas” Rosa, Anita, Ele y Belén, todas diferentes y todas incomparables.
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