Epidemiología Y Control De La Fiebre Q (Coxiella Burnetii) En Fauna Silvestre Ibérica

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Epidemiología Y Control De La Fiebre Q (Coxiella Burnetii) En Fauna Silvestre Ibérica Epidemiología y control de la fiebre Q (Coxiella burnetii) en fauna silvestre ibérica. David González Barrio Tesis doctoral 1 Epidemiología y control de la fiebre Q (Coxiella burnetii) en fauna silvestre ibérica. Trabajo de investigación presentado por David González Barrio para optar al grado de Doctor por la Universidad de Castilla-La Mancha Ciudad Real, 2015 Grupo de Sanidad y Biotecnología (SaBio) Instituto de Investigación en Recursos Cinegéticos (IREC; CSIC- UCLM-JCCM) Departamento de Ciencia y Tecnología Agroforestal y Genética Universidad de Castilla-La Mancha 2 Los abajo firmantes, como directores de este tesis doctoral, hacemos constar que la Tesis titulada “Epidemiología y control de la fiebre Q (Coxiella burnetii) en fauna silvestre ibérica”, y realizada por David González Barrio, reúne los requisitos necesarios para su defensa y aprobación y, por tanto, para optar al grado de doctor con mención internacional. Vº Bº de los Directores Dr José Francisco Ruiz Fons Dr Isabel G. Fernández de Mera Dr Christian Gortázar Schmidt 3 La realización de este trabajo ha sido posible gracias a las siguientes entidades y proyectos de investigación: Fundación de la Universidad de Castilla-La Mancha Cátedra Fundación Enresa Proyecto Europeo Strategies For The Eradication Of Bovine Tuberculosis Proyecto Europeo, Comisión Europea Harmonised Approaches In Monitoring Wildlife Population Health, And Ecology And Abundance (APHAEA) Proyecto Europeo, Comisión Europea (VII Programa Marco) ANTIcipating the global onset of new epidemics (ANTIGONE) Junta de Comunidades de Castilla-La Mancha Estructura de los contactos y riesgo de transmisión de enfermedades entre Ganado y ungulados silvestres CGT – Apoyo Al Desarrollo De Nuevas Tecnologias Para El Control Sanitario de La Fauna Silvestre Ministerio de Economía y Competitividad Centro para el Desarrollo Tecnológico Industrial (CDTI) – Incorporación de Nuevas Metodologías para la tecnificación y sostenibilidad de explotaciones bovinas extensivas y cinegáticas. Financiación Adicional Del Contrato Ramón Y Cajal De José Francisco Ruiz Fons 4 Resumen. Capítulo I: Introducción general. Capítulo II: Epidemiología de Coxiella burnetii en fauna silvestre ibérica. 1. Estado de Coxiella burnetii en las poblaciones de ciervo rojo (Cervus elaphus) en la península ibérica y factores de riesgo asociados. “Host and Environmental Factors Modulate the Exposure of Free-Ranging and Farmed Red Deer (Cervus elaphus) to Coxiella burnetii” 2. Estado de Coxiella burnetii en las poblaciones de conejo de monte (Oryctolagus cuniculus) en la península ibérica y factores de riesgo asociados. “European Rabbits as Reservoir for Coxiella burnetii” 3. Dinámica de la infección por Coxiella burnetii en una población endémica de ciervo en condiciones semi-extensivas. “Long-term dynamics of Coxiella burnetii in farmed red deer (Cervus elaphus)” 4. Genotipos de Coxiella burnetii presentes en fauna silvestre en la península ibérica basados en MLVA. “Coxiella burnetii genotypes in Iberian wildlife” 5. Genotipado de Coxiella burnetii de fauna silvestre ibérica mediante PCR e hibridación RLB y relaciones con genotipos de ganado doméstico y humanos en España. “Coxiella burnetii genotypes in Spanish wildlife: implications for livestock and human health” 5 Capítulo III: Vías de transmisión de Coxiella burnetii en fauna silvestre ibérica. 1. Vías de excreción de Coxiella burnetii y otros patógenos relevantes en jabalí (Sus scrofa). “Shedding patterns of endemic Eurasian wild boar (Sus scrofa) pathogens” 2. Vías de excreción de Coxiella burnetii en ciervo rojo (Cervus elaphus) en condiciones de producción semi-extensiva. “Coxiella burnetii Shedding by Farmed Red Deer (Cervus elaphus)” Capítulo IV: Estrategias de control de Coxiella burnetii: evaluación de la vacunación con vacunas inactivadas comerciales de fase I como estrategia de reducción de la prevalencia y el nivel de excreción de la bacteria en ciervo rojo (Cervus elaphus). Capítulo V: Síntesis general y conclusiones. 6 Estructura de la tesis La finalidad de este trabajo de investigación consiste en estudiar el papel de la fauna silvestre en la epidemiología de Coxiella burnetii y en evaluar posibles medidas de control - que al igual que en especies domésticas, sobre todo rumiantes - pueden llevarse a cabo para disminuir la contaminación ambiental - fuente de infección para otras especies silvestres, domésticas y para las personas - y, así, la transmisión. La presente Tesis Doctoral se estructura en una primera sección (Resumen) que pretende plasmar de forma resumida los aspectos metodológicos y resultados más destacables obtenidos en cada capítulo. A continuación se presenta el estado actual del tema (Capítulo I) a modo de Introducción General a los trabajos de la Tesis Doctoral, en la que se aborda el conocimiento general sobre C. burnetii, patógeno intracelular obligado con un amplio rango de hospedadores y con carácter zoonósico que lo convierte en la causa de brotes epidémicos de fiebre Q en personas. Esta introducción abarca aspectos generales del patógeno (etiología, estructura genética, etc) y de sus efectos en hospedadores donde la epidemiología y el resultado de la infección han sido bien caracterizados (principalmente rumiantes domésticos y ser humano), pero su intención fundamental es centrarse en el conocimiento existente sobre el papel que juega la fauna silvestre en la epidemiología de C. burnetii y en las estrategias de control potencialmente aplicables en fauna silvestre. Incluye aspectos relacionados con C. burnetii en fauna silvestre como las particularidades de su diagnóstico, desde la detección serológica de anticuerpos frente a C. burnetii a la caracterización molecular de los genotipos circulantes en diversas especies silvestres a nivel mundial y la relación filogenética con los genotipos aislados en otras especies de hospedadores, así como el conocimiento existente sobre los factores de riesgo de mantenimiento y transmisión de C. burnetii asociados a la fauna silvestre. También en esta introducción se abordan 7 aspectos de la transmisión, patogenia y patología conocidos en fauna silvestre, así como el conocimiento actual sobre los métodos de control de las enfermedades en la fauna silvestre. El Capítulo II se centra en el abordaje científico llevado a cabo en esta Tesis Doctoral para mejorar el conocimiento de la epidemiología de C. burnetii en poblaciones de fauna silvestre ibérica, en concreto en el ciervo rojo (Cervus elaphus) y en el conejo de monte (Oryctolagus cuniculus), así como para identificar algunos de los factores de riesgo que determinan la transmisión y mantenimiento de C. burnetii en un ciclo silvestre en estas especies. En este capítulo también se aborda un estudio que pretende comprender la dinámica temporal de C. burnetii en poblaciones de ciervo rojo endémicas, usando una granja ibérica de ciervo rojo como modelo. Dos estudios adicionales abordan el tipado molecular de las cepas de C. burnetii circulantes en especies silvestres ibéricas. En estos estudios se compara mediantes dos técnicas de tipado molecular diferentes (MLVA y PCR-RLB) los genotipos que circulan en la fauna silvestre ibérica con cepas presentes en ganado doméstico y en casos clínicos humanos en España y en el resto del mundo. En el siguiente capítulo (Capítulo III) se aborda el estudio de las vías de transmisión de C. burnetii en diferentes especies de fauna silvestre ibérica, concretamente en el jabalí (Sus scrofa) y en el ciervo rojo (Cervus elaphus), así como las potenciales implicaciones clínicas de la infección por C. burnetii en el ciervo. El último de los estudios científicos que componen la presente Tesis Doctoral (Capítulo IV) aborda el diseño y aplicación de estrategias potenciales de control de C. burnetii en ciervo usando como modelo una granja de ciervo ibérica. El control de C. burnetii en rumiantes domésticos se basa principalmente en el uso de vacunas inactivadas de bacterias en fase I con la finalidad de reducir la prevalencia de excreción y, en paralelo, los niveles de excreción para reducir la transmisión y, con ello, la incidencia. Este capítulo 8 se basa en el diseño de un programa de vacunación experimental en condiciones de campo y en la implementación de una vacuna comercial inactivada de C. burnetii en fase I (COXEVAC, CEVA Santè Animale, Francia), así como en la posterior evaluación de la eficacia de la vacuna en inducir inmunidad humoral y en reducir la excreción de C. burnetii en secreciones vaginales, leche y heces en un periodo de tres años desde la implementación del programa vacunal. Esta Tesis Doctoral se cierra con una Síntesis General (Capítulo V) y Conclusiones en la que se destacan los principales hitos logrados, la aplicabilidad de los resultados y las necesidades futuras de abordaje científico necesarias para mejorar el conocimiento sobre la epidemiología, la patogenia, la clínica y el control de C. burnetii en la fauna silvestre. Esta sección incluye las conclusiones derivadas del trabajo de investigación desarrollado en los diferentes estudios que componen la Tesis Doctoral. 9 Introducción 1. El patógeno y sus características Coxiella burnetii, previamente denominada como Rickettsia diaporica o Rickettsia burnetii es una bacteria que se encuadra dentro de la clase Gammaproteobacteria, orden Legionellales, familia Coxiellaceae. Es un bacilo gram negativo, patógeno intracelular obligado, pequeño (0.2-0.4 μm ancho, 0.4-1.0 μm de largo), no capsulado y pleomórfico (Maurin y Raoult, 1999). Coxiella burnetii presenta dos formas antigénicas
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