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Contrib Aused B Universitat Autònoma de Barcelona Facultat de Veterinària Departament de Biologia Animal, de Biologia Vegetal i d’Ecologia Contributions to the knowledge of a new disease caused by an amoeba in ongrowing Senegalese sole, Solea senegalensis (Kaup 1858) Tesiss Doctoral Memoria de tesis doctoral presentada por María Constenla Matalobos para optar al grado de Doctora en Acuicultura, realizada bajo la codirección del Dr. Francesc Padróós i Bover de la Universitat Autònoma de Barcelona y del Dr. Oswaldo Palenzuela Ruiz del Instituto de Acuicultura de Torre la Sal (CSIIC) La presente tesis doctoral está adscrita al doctorado de Accuicultura. Director Director Dr. FRANCESC PADRÓS i BOVER Dr. OSWALDO PALENZUELA RUIZ Doctoranda MARIA CONSTTENLA MATALOBOS Barcelona, diciembre 2012 Con el apoyo de una beca predoctoral de la Universitat Autònoma de Barcelona (PIF) Parte del estudio experimental se ha realizado en el Instituto de Acuicultura de Torre la Sal (IATS-CSIC) y se ha financiado parcialmente por el Ministerio Español de Ciencia e Innovación y por las empresas de acuicultura a través de proyectos internos de los programas de investigación del CSIC (intramuros). Financiación adicional también ha sido otorgada por el Gobierno regional (Generalitat Valenciana PROMETEO 2010/006 y la CIIU 2012/003). Impossible is just a big word thrown around by small men who find it easier to live in the world they’ve been given, than to explore the power they have to change it. Impossible is not a fact, it’s an opinion. Impossible is not a declaration, it’s a dare. Impossible is potencial. Impossible is temporary. Impossible is Nothing. ‘The soul of a butterfly’ Muhammad Ali. A mis padres y a Edu Acknowledgements Primero de todo quisiera agradecer a mis directores todo el apoyo que me habéis transmitido durante estos años, y muy especialmente en esta locura de apretón final. Sin vosotros está claro que no lo hubiese conseguido. También me gustaría agradecer concretamente a Sito, por la oportunidad de crecer como profesional a su lado y, en cierta medida, también como persona y de brindarme la oportunidad de ser de alguna manera su pequeña padawan. Y a Oswaldo por introducirme con paciencia en el “fascinante” mundo de la biología molecular. Chicos, ha sido un inmenso honor poder trabajar a vuestro lado durante estos 4 años. En este apartado dedicado a mis directores, no me gustaría olvidarme de Paco, director de la tesis durante un tiempo y gran persona y compañero. Gracias por llenarme la cabeza de histerotilaciums, rafidascaris, apéndices ventriculares y ciegos intestinales, ahora mi cabeza ya me pesa un poquito más. Me gustaría continuar agradeciendo a todo el personal de las granjas que han colaborado con nosotros de alguna u otra forma. Aunque me gustaría nombraros uno por uno, la confidencialidad de los datos no me lo permite. Vosotros ya sabéis quienes sois y espero que os deis por aludidos. Os estoy muy muy muy agradecida, ya que sin vuestro apoyo este trabajo no tendría sentido ni razón de ser. También me gustaría agradecer a mis compañeros de departamento. A Silvia Crespo, porque gracias a ella y a su asignatura de Patología de acuáticos empecé a interesarme por este maravilloso mundo. A Maite Carrassón, por abrirme las puertas del departamento. A Queta y a Marta, sin las cuales nada hubiese sido lo mismo ya que en vosotras he encontrado no solo compañeras de trabajo sino amigas de verdad. A Queta, no solo le agradezco los innumerables buenos momentos que me ha regalado durante las horas de trabajo, sino los otros múltiples en la vida personal, convirtiéndose en una persona muy importante para mí. Por poner un ejemplo divertido y dejar la lagrimilla para otra ocasión, me gustaría mencionar las noches de desestrés y bailoteo en ese maravilloso garito nocturno con nombre de crustáceo donde “Carlos” siempre nos ha cuidado poniéndonos los mejores Acknowledgements temazos, por mucho que se cierren ciclos no hay que dejar las buenas costumbres. Y por supuesto, me gustaría agradecer todos los momentos y apoyo al resto de mis compañeros con los que he tenido la suerte de compartir el día a día: Encarna, por ser la “mami” que siempre se preocupa y nos cuida, a Roger, por su buenrollismo, a Ana por sus consejos y a Sandra, a la que tanto he mareado la cabeza con los cortes histológicos y sin la cual las hibridaciones de esta tesis no hubiesen salido tan bien. Y por supuesto, a Montse y a los dos Alberts que, aunque ya hace tiempo que no nos visitan, me acompañaron en el principio de esta aventura. Sin olvidarme también de tod@s las alumn@s colaboradores que han ido pasando por el departamento, y que han dado un poco de alegría a las tardes de disección. Gracias también a las niñas y el resto de compañeros de la unidad zoología, que aunque parece que estamos muy alejados, siempre se agradecen los encuentros para compartir unas risas. Y por supuesto, no me puedo olvidar de los múltiples superhéroes y dioses que siempre pueden aparecer por el departamento: BATMAN, Silversurfer, Skakermen, y últimamente Atlas, siempre soportando el gran peso del conocimiento del mundo; de todos ellos he podido aprender a ver la vida desde diferentes perspectivas. También me gustaría agradecer al equipo de microscopía electrónica de la UAB, especialmente a Álex y a Paqui, por su paciencia durante la búsqueda de parásitos y orgánulos diminutos, y por la explicaciones y revisiones de todos los protocolos. En el Instituto de Acuicultura de Torre la Sal (IATS) he realizado prácticamente la mitad de este trabajo, y podría dedicar hojas y hojas de agradecimientos, pero voy a intentar resumir agradeciendo al todo el equipo del IATS en general y especialmente al grupo de patología: A Pilar Álvarez- Pellitero y a Ariadna Sitjà-Bobadilla por sus observaciones y consejos desde el principio del planteamiento de esta tesis, a MªJosé por ayudarme con las hidridaciones, a Gregorio por ayudarme con los cultivos, y a Raquel, María e Itziar por toda la paciencia y buen humor con mis: ¿esto donde está? no encuentro lo otro, ¿se han acabado las puntas? ¿me puedes pasar el Acknowledgements protocolo? tengo que poner un autoclave…en fin..muchas gracias por estar siempre ahí. Pero también le agradezco al resto de grupos, a Oli, Gabi, Diana, David… por acogerme desde el principio como una más. Además, en este apartado de Castellón, me gustaría hacer un agradecimiento especial a mi familia castellonense, especialmente a Quique e Itziar, Lucía e Inma, por abrirme las puertas de su casa y dejarme ser una más de la familia. Sin duda mis múltiples estancias en Castellón no hubiesen sido lo mismo sin vosotras/os: los zumitos recién exprimidos por la mañana, las nuevas y exquisitas recetas de comida, los conciertillos, las cañitas (aunque cada vez tuviésemos menos tiempo para el ocio), gracias por no cerrarme la puerta al llegar tarde de currar, por esperarme con una sonrisa e incluso con la cena hecha!!! Siempre acompañado de muy buena música e, incluso en ocasiones, de conciertos en vivo y en “petit comité” de los niños del swing ;) Al grupo de AP de la facultad de Veterinaria de Lugo: Maribel, Roberto, Dani, Ana, Meli, Sonia…porque me enseñasteis las bases de la anatomía patológica cuando aun estudiaba veterinaria, que me fascinó, y me abristeis las puertas de vuestro laboratorio para poder participar en lo que fueron mis primeros muestreos en peces. Lot of thanks to the Institute of Parasitology Academy of Sciences of Ceské Budejovice (Czech Republic), especially to Iva Dyková, for their immense hospitality and kindness during my stay in her lab and also for his good advices and suggestions. Also thanks to Dr. Ivan Cepicka (Department of Zoology, Charles University of Prague) for their suggestions. DČkuju. Y por supuesto, no me puedo olvidar del apoyo y la buena voluntad de algunas de las personas que me he ido encontrando en el camino, sin las cuales este trabajo tampoco hubiese sido lo mismo. Me gustaría agradecer al ECIMAT, especialmente a Antonio G. Villlanueva, por el apoyo que nos ha brindado en la última etapa. A Carlos Zarza por su continuo apoyo, interés y ánimos, aunque ahora se encuentre al otro lado del mundo. Y a Daniel Gijón por estar siempre ahí para echar una mano. Acknowledgements Y por último, pero no menos importante, a mi familia, muy especialmente a mis padres, por su apoyo incondicional siempre y en todo momento, aunque no tengan muy claro si estudio, trabajo o a que me dedico exactamente. Y a Edu, por enseñarme que impossible is nothing, y por mimarme y cuidarme siempre y tener una paciencia infinita. Espero no haberme olvidado de nadie, si es así lo siento muchísimo. Por todo esto y mucho más, a cada uno de vosotr@s, ¡¡GRACIAS INFINITAS!! Abstract Abstract: A previously undescribed pathological condition is affecting the culture of Solea senegalensis in some farms of the Atlantic coast of Spain. This condition is characterised by the presence of external protuberances in the skin of the affected fish. These lesions correspond to nodules in the muscular tissue showing an abscess-like aspect. Similar lesions were found in kidney, heart, liver and the digestive tract, which leads us to define this pathology as a systemic disease. Histological sections of these nodules revealed the presence of a large core formed mainly of necrotic tissue surrounded with fibroblasts and macrophages. Round-shaped plasmodial organisms were found in the external layer of the nodules and usually inside macrophages or fibroblasts. These organisms were also observed in the intestinal mucosa and submucosa, without causing apparent lesions. This organisms are correspond to a new amoeba species, that belongs to the family Entamoebidae (Phylum Amebozoa, Infraphylum Archamoeba), and we tentatively describe it as a new species in the genus Endolimax, Endolimax piscium n.
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