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Caracterización Genotípica Y Quimiotaxonómica De Especies Termofílicas De Campylobacter Aisladas De Carne De Ave

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Caracterización Genotípica Y Quimiotaxonómica De Especies Termofílicas De Campylobacter Aisladas De Carne De Ave UNIVERSIDAD DE LEÓN DEPARTAMENTO DE HIGIENE Y TECNOLOGÍA DE LOS ALIMENTOS PREVALENCIA , CARACTERIZACIÓN QUIMIOTAXONÓMICA MEDIANTE ESPECTROSCOPÍA DE INFRARROJOS E IDENTIFICACIÓN CON REDES NEURONALES ARTIFICIALES Y ANÁLISIS MULTIVARIANTE DE ESPECIES TERMOFÍLICAS DE CAMPYLOBACTER AISLADAS DE AVES Y PRODUCTOS AVÍCOLAS Juan Prieto Gómez 2 INFORME DEL DIRECTOR DE LA TESIS (Art. 11.3 del R.D. 56/2005) El Dr. Miguel Prieto Maradona, como Director de la Tesis Doctoral titulada “Prevalencia, caracterización quimiotaxonómica mediante espectroscopía de infrarrojos e identificación con redes neuronales artificiales y análisis multivariante de especies termofílicas de Campylobacter aisladas de aves y productos avícolas”, de la que es autor D. Juan Prieto Gómez, realizada en el Departamento de Higiene y Tecnología de los Alimentos de la Universidad de León, informa favorablemente el depósito de la misma, dado que reúne las condiciones necesarias para su defensa, y suscribiéndolo para dar cumplimiento al art. 11.3 del R.D. 56/2005, en León a 16 de Abril de 2010 Fdo. Miguel Prieto Maradona 3 4 ADMISIÓN A TRÁMITE DEL DEPARTAMENTO (Art. 11.3 del R.D. 56/2005 y Norma 7ª de las Complementarias de la ULE) El Departamento de Higiene y Tecnología de los Alimentos en su reunión celebrada el día 28 de Septiembre de 2010 ha acordado dar su conformidad a la admisión a trámite de lectura de la Tesis Doctoral titulada “Prevalencia, caracterización quimiotaxonómica mediante espectroscopía de infrarrojos e identificación con redes neuronales artificiales y análisis multivariante de especies termofílicas de Campylobacter aisladas de aves y productos avícolas”, dirigida por el Dr. D. Miguel Prieto Maradona, elaborada por D. Juan Prieto Gómez y cuyo título en inglés es el siguiente “Prevalence, chemotaxonomic characterization using infrared spectroscopy and identification with artificial neural networks and multivariate analysis of termophilic Campylobacter spp. isolated from poultry and poultry products”. Lo que firmo, para dar cumplimiento al art. 11.3 del R.D. 56/2005, en León a 28 de Septiembre de 2010. El Secretario, Fdo.: Mª Rosario García Armesto Vº Bº El Director del Departamento, Fdo.: Carlos Alonso Calleja 5 6 Esta tesis ha sido llevada a cabo con las ayudas económicas del Fondo de Investigaciones Sanitarias (FIS), gracias al proyecto "Monitorización epidemiológica e identificación clínica de poblaciones de Campylobacter responsables de toxiinfecciones alimentarias mediante el análisis de su espectro infrarrojo" (Ref. 00/0930), y de la Junta de Castilla y León, con el proyecto “Identificación y tipado de microorganismos patógenos causantes de toxiinfecciones alimentarias: comparación del espectro infrarrojo y las secuencias del RNA ribosómico” (Ref. LE09-98), y cuyo investigador principal es el Dr. Miguel Prieto Maradona. Asimismo, para la elaboración de este trabajo, D. Juan Prieto Gómez ha disfrutado de una beca de la Fundación MAPFRE MEDICINA entre febrero de 2001 y febrero de 2002. Igualmente, para la elaboración de esta Tesis Doctoral, D. Juan Prieto Gómez ha obtenido financiación del Centro de Estudios e Investigación para la Gestión de Riesgos Agrarios y Medioambientales (CEIGRAM) en su convocatoria de ayudas a jóvenes investigadores de 2010. 7 8 AGRADECIMIENTOS Es probable que sean estas las líneas más difíciles de redactar de todo este trabajo. No ya sólo por no olvidar a nadie de quienes, en mayor o menor medida, han colaborado y me han ayudado en la elaboración de esta Tesis Doctoral, si no por la imposibilidad de plasmar mediante la palabra la enorme carga emocional que supone el trabajo, las satisfacciones, las preocupaciones, los éxitos y los fracasos de todos estos años. Reflejar por escrito tal cúmulo de sensaciones se me antoja una labor imposible de abordar, aún a pesar de contar con la valiosísima herramienta del lenguaje libre –que no literario-, mucho más preciso que el científico a la hora de alcanzar este objetivo. Además, a lo largo de todo este tiempo me he ido dando cuenta de que realizar una Tesis Doctoral trasciende del ámbito científico y profesional, para convertirse en una de las, seguramente, más importantes vivencias de aquellos que nos hemos embarcado en semejante aventura. Es por ello justo y obligado, y a la vez un orgullo y un honor, recordar en este momento a todos aquellos que me han ayudado a alcanzar el lugar en el que hoy me encuentro. En primer lugar a mi Director, el Dr. Prieto Maradona. Miguel me brindó su confianza el primer día y la ha mantenido en todo momento, incluso cuando los obstáculos parecían demasiados y mi camino profesional se orientaba por otras sendas alejadas de la investigación. Su comprensión y su apoyo ante una decisión tan trascendente en mi vida profesional constituyeron el más valioso de los estímulos para concluir, a pesar de las dificultades subsiguientes a asumir, un trabajo iniciado y en todo caso muy querido. Espero haber sido capaz de devolverle esa misma confianza y haber adquirido de él su rigor y su capacidad de análisis profundo con los que me ha formado estos años. Y, ante todo, le agradezco que la relación forjada en este tiempo haya ido más allá de la del maestro con su alumno y se haya transformado en una sólida amistad. A los miembros del Departamento de Higiene y Tecnología de los Alimentos de la Universidad de León. Todos ellos, profesores y personal de Administración y Servicios, me han brindado su colaboración en todo momento y me han ayudado a resolver las innumerables dudas y problemas que se presentan al enfrentarse a una Tesis Doctoral. Y lo han hecho, además, siempre con una sonrisa y una palabra amable, que espero haberles sabido corresponder. 9 Al resto de becarios que lo fueron conmigo. Compañeros de trabajo, de algunos momentos de malestar y de muchos más momentos de risas dentro y fuera del laboratorio. Quisiera nombrarlos en orden alfabético, única manera de hacerles justicia: Chema, Daniel, Fanny, Irma – nuestra gran maestra –, Isabel, Oriol, Óscar, Toño y Víctor. A quienes me han ayudado fuera de los límites de mi querida Universidad de León, en mis estancias en las Universidades de Gante y Varsovia. Las enseñanzas, las experiencias y las vivencias compartidas por muchos de quienes, procedentes de distintas Universidades de toda Europa, me acompañaron allí, me han servido de ayuda y estímulo en numerosas ocasiones a lo largo de estos años. Al personal de las granjas y mataderos visitados para el muestreo de este trabajo, y, de manera especial, a sus profesionales veterinarios por las enseñanzas e informaciones tan generosamente brindadas. Sin embargo, todo el apoyo profesional y académico nunca es suficiente para ayudar a encarar las dificultades con las que se presenta una Tesis Doctoral, y, así, el empuje, los ánimos y la ilusión compartida de los más cercanos han resultado imprescindibles para llevarme a este momento. Familia, amigos y compañeros han sabido dar el aliento necesario en el momento preciso. Y entre ellos, sin duda alguna, Silvia y mi familia, de quienes confío que esta Tesis les haga seguirse sintiendo orgullosos de mí. Silvia, ejemplo de tantas cosas y, en el ámbito académico, pese a proceder de un campo del saber humano muy diferente, por su espíritu crítico y por su rigor, pero compatibilizando en todo momento su exigencia intelectual con su dulzura y amor. Llegó en los últimos pasos de esta aventura, pero me hace enormemente feliz que pueda ser testigo de su final. Mi hermana, Julia y, con ella, su Julio. Agradezco todo el cariño de la mejor de las hermanas: compañera y consejera sagaz y brillante. Por ambos siento una profunda admiración, por sus valores y principios, referencia constante en mi trayectoria. 10 Mis padres, Bernardino y María Teresa. Es un esfuerzo vano intentar siquiera resumir las razones de mi agradecimiento hacia ellos, tantas, tan diversas y tan profundas. La ayuda de mis padres – y refiriéndome tan solo a lo estrictamente profesional – ha sido imprescindible en todo momento. Lo fue durante mi formación como Veterinario y en mi licenciatura en Ciencia y Tecnología de los Alimentos, así como en mis primeros pasos profesionales y en mi ingreso en el Cuerpo Nacional Veterinario. Y, desde luego, lo ha sido durante la elaboración de esta Tesis Doctoral. Y por último, a la Profesión y a la Ciencia Veterinaria, vieja, noble y sabia profesión. Hijo como soy de veterinarios, continuador de una saga veterinaria que se adentra hasta mediados del siglo XIX, la Veterinaria está tan indefectiblemente unida a mí y a mi familia, que la llevo no sólo en el corazón, si no hasta en mi propia dotación genética. Ojalá este esfuerzo de estos años contribuya, siquiera mínimamente, a hacerla aún más grande, y sirva este trabajo de agradecimiento a la Veterinaria por todo lo que significa en mi vida. Muchas gracias Juan Prieto Gómez 11 12 El sueño de la razón produce monstruos Francisco de Goya y Lucientes 13 14 A los míos: a quienes fueron; a quienes son; a quienes hayan de ser. 15 16 ÍNDICE ÍNDICE ____________________________________________________________ 17 1 Capítulo I. Introducción __________________________________________ 21 1.1 Historia _________________________________________________________ 22 1.2 Fisiología y taxonomía del género Campylobacter _______________________ 23 1.3 Aislamiento y detección de Campylobacter spp. ________________________ 24 1.4 Pruebas fenotípicas y genotípicas en identificación y tipificación ___________ 25 1.4.1 Métodos fenotípicos ____________________________________________________ 25 1.4.2
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