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Estrategias De Transmisión De Digeneos Naturalis Repositorio Institucional Universidad Nacional de La Plata http://naturalis.fcnym.unlp.edu.ar Facultad de Ciencias Naturales y Museo Estrategias de transmisión de digeneos parásitos que utilizan gasterópodos como hospedadores intermediarios en la costa patagónica Gilardoni, Carmen Doctor en Ciencias Naturales Dirección: Cremonte, Florencia Co-dirección: Díaz, Julia I. Facultad de Ciencias Naturales y Museo 2014 Acceso en: http://naturalis.fcnym.unlp.edu.ar/id/20140424001339 Esta obra está bajo una Licencia Creative Commons Atribución-NoComercial-CompartirIgual 4.0 Internacional Powered by TCPDF (www.tcpdf.org) Facultad de Ciencias Naturales y Museo Universidad Nacional La Plata ESTRATEGIAS DE TRANSMISIÓN DE DIGENEOS PARÁSITOS QUE UTILIZAN GASTERÓPODOS COMO HOSPEDADORES INTERMEDIARIOS EN LA COSTA PATAGÓNICA Trabajo de Tesis para optar por el título de Doctor en Ciencias Naturales Lic. Carmen Gilardoni Directora: Dra. Florencia Cremonte Co-directora: Dra. Julia I. Diaz 2013 AGRADECIMIENTOS Todas las etapas de la vida tienen un final. Y los finales felices se alcanzan con voluntad, esfuerzo, constancia, motivación y con ayuda de muchas personas que te acompañan en el camino. Por eso quiero agradecer…. A mi directora Florencia Cremonte, por su apoyo incondicional, por la motivación, por darme alas para crecer en la investigación, por su amistad. A mi codirectora Julia Diaz, por su predisposición y entusiasmo, por estar presente siempre que la necesité. Al Dr. Cristián Ituarte, por su objetividad y ejemplo, por ayudarme en numerosas instancias, en las colectas y en los trabajos de laboratorio, por enseñarme a realizar cortes histológicos y por enriquecer con sus conocimientos este trabajo. Los cortes histológicos se realizaron en la División Zoológica de Invertebrados, Museo Argentino de Ciencias Naturales, en el marco de una pasantía dirigida por el Dr. Ituarte. A mis compañeras de trabajo, Gise, Emi, Nuri y Estefa, por colaborar conmigo en diferentes etapas de este trabajo (colectas, búsqueda de bibliografía, trabajos de laboratorio), darme ánimo y por mantener siempre el espíritu de equipo. Al Centro Nacional Patagónico (CENPAT-CONICET) y especialmente al Laboratorio de Parasitología, por permitirme desarrollar todas mis actividades de doctorado y brindarme los espacios y herramientas para el desarrollo de esta tesis. A ALUAR S.A. por brindarnos el servicio de Microscopía Electrónica de Barrido y especialmente a Jaime Groizard que siempre nos recibe con mucho entusiasmo y se esfuerza para que las fotografías queden perfectas. A los Dres. Pedro Rodrigues y Susana Pina por enseñarme sobre técnicas moleculares y por brindarme su amistad. Los estudios moleculares se realizaron en el Laboratorio de Sanidad e Inmunología del Instituto de Ciencias Biomédicas Abel Salazar (ICBAS), Universidad de Porto, Portugal, en colaboración con el Dr. Rodrigues y la Dra. Pina, en el marco de una beca Eurotango otorgada por la Fundación Erasmus Mundus. Al Dr. Vasyl Tkach por realizar los estudios moleculares de una especie descripta en este estudio. Las larvas y adultos de Parorchis sp., fueron secuenciadas por el Dr. Tkach, Universidad de Dakota del Norte, Estados Unidos, en colaboración con el grupo de trabajo. Al Dr. Andrés Averbuj por el material provisto para los estudios moleculares. A los Dres. Atila Goztonyi y David Galván por brindarme información sobre los peces de arrecife. A los Dres. Luis Bala, Ana Millones y a la Mg. Annick Morgenthaler por brindarme información sobre aves marinas. A los Dres. Diego Zelayas, Gregorio Bigatti, Federico Márquez y Fabrizio Scarabino por brindarme información sobre los moluscos. Al. Dr. Hermes Mianzán por la información provista sobre ctenóforos. A todos aquellos que me acompañaron y ayudaron en los muestreos, Mica, Joaco, Ema, Estefa, Gise y Cristián. A los jurados de esta tesis, Dra. Verónica Flores, Dres. Sergio Martorelli y Néstor Ciocco por sus sugerencias que contribuyeron a mejorar el manuscrito. A mi familia, padres, hermanos, tíos, sobrinos, primos y amigos de la vida, que fueron mi principal motivación y mi fuerza para siempre seguir adelante. A mi papá Carlos, por su confianza, sus consejos, su protección, por todo lo que me enseñó y me acompaño durante toda mi vida. A mi mamá Rosita, por su paciencia, su entendimiento, su amor de madre, por estar siempre presente en mi vida. A mi hermana Luján, por su alegría y entusiasmo, por ser mi amiga y darme el regalo más lindo del mundo, mi sobrino ahijado Lucca. A mi hermano Marcelo, por su fortaleza, sus charlas y su sinceridad. A mis hermanas de la vida, Gise, Cintia, Lucía, Lau y Flor por estar en las buenas y en las malas. A mis suegros Elba y Miguel, por cuidarme y quererme como una hija propia A Mica, Mauricio y Vicente, por llenar mi vida de alegría y amor estos últimos años. INDICE RESUMEN………………………………………………………………………………….1 ABSTRACT………………………………………………………………………………...5 INTRODUCCIÓN GENERAL Y OBJETIVOS………………………………………...9 MATERIALES Y MÉTODOS GENERALES………………………………………….13 Sitios de estudio…………………………………………………………………………...13 Hospedadores……………………………………………………………………………..17 Colecta de los hospedadores..............................................................................................21 CAPÍTULO 1: DESCRIPCIÓN DE LOS DIGENEOS LARVALES………………..22 1.1. Introducción…………………………………………………………………………..22 1.2. Materiales y métodos………………………………………………………………...24 1.2.1. Procesamiento y estudio morfológico de los estadios larvales………………...24 1.2.2. Estudios moleculares………………………………………………………………26 1.3. Resultados…………………………………………………………………………….32 Hemiuroidea………………………………………………………………………………32 Hemiuroidea fam. gen. et sp……………………………………………………..32 Lepocreadiidae……………………………………………………………………………38 Lepocreadiidae gen. et sp. 1……………………………………………………...38 Lepocreadiidae gen. et sp. 2……………………………………………………...45 Zoogonidae………………………………………………………………………………...53 Diphterostomum sp………………………………………………………………....53 Zoogonus sp………………………………………………………………………...60 Gymnophallidae…………………………………………………………………………..63 Gymnophalloides nacellae…………………………………………………………...63 Microphallidae…………………………………………………………………………….68 Maritrema sp. 1……………………………………………………………………..68 Maritrema madrynense……………………………………………………………..75 Notocotylidae……………………………………………………………………………...82 Notocotylidae gen. et sp………………………………………………………….82 Renicolidae………………………………………………………………………………...86 Renicolidae gen. et sp. 1…………………………………………………………..86 Renicolidae gen. et sp. 2…………………………………………………………..92 Philophthalmidae…………………………………………………………………………96 Parorchis sp…………………………………………………………………………96 Schistosomatidae………………………………………………………………………...103 Schistosomatidae gen. et sp……………………………………………………..103 1.4. Discusión general y conclusiones…………………………………………………109 CAPÍTULO 2: CICLOS DE VIDA DE LOS DIGENEOS…………………………..113 2.1. Introducción…………………………………………………………………………113 2.2. Materiales y métodos……………………………………………………………….120 2.2.1. Búsqueda bibliográfica…………………………………………………………...120 2.2.2. Estudio de potenciales segundos hospedadores intermediarios…………….120 2.2.3. Infecciones experimentales………………………………………………………121 2.2.4. Cultivos in vitro……………………………………………………………………122 2.2.5. Estudios moleculares……………………………………………………………..122 2.3. Resultados y discusión……………………………………………………………...124 2.3.1. Digeneos que finalizan su ciclo de vida en peces……………………………...124 2.3.1.1. Hemiuroidea fam. gen. et sp…………………………………………………..124 2.3.1.2. Lepocreadiidae gen. et sp. 1 y Lepocreadiidae gen. et sp. 2………………..130 2.3.1.3. Diphterostomum sp. y Zoogonus sp. (Zoogonidae)……………………………135 2.3.2. Digeneos que finalizan su ciclo de vida en aves……………………………….140 2.3.2.1. Gymnophalloides nacellae (Gymnophallidae)…………………………………..140 2.3.2.2. Maritrema sp. 1 y Maritrema madrynense (Microphallidae)…………………..143 2.3.2.3. Notocotylidae gen. et sp………………………………………………………..153 2.3.2.4. Renicolidae gen. et sp. 1 y Renicolidae gen. et sp. 2…………………………156 2.3.2.5. Parorchis sp. (Philophthalmidae)………………………………………………163 2.3.2.6. Schistosomatidae gen. et sp…………………………………………………….166 2.4. Discusión general y conclusiones………………………………………………….168 CAPÍTULO 3: DINAMICA ESPACIAL Y ESTACIONAL DE LAS POBLACIONES Y COMUNIDADES DE LOS DIGENEOS ESTUDIADOS…...180 3.1. Introducción………………………………………………………………………….180 3.2. Materiales y métodos……………………………………………………………….182 3.2.1. Estudios descriptivos y análisis estadísticos de la prevalencia en función de la talla y el sexo del hospedador, la estación del año y el sitio de estudio……………182 3.2.2. Análisis de las comunidades de digeneos en los dos sitios de estudio……...183 3.3. Resultados…………………………………………………………………………...186 3.3.1. Digeneos larvales del intermareal de Puerto Madryn………………………...186 3.3.2. Digeneos larvales del intermareal de Puerto Deseado………………………..191 3.3.3. Comparación de la prevalencia y diversidad de las comunidades de digeneos en los intermareales de Puerto Madryn y Puerto Deseado………………………….201 3.4. Discusión y conclusiones………………………………………………………….. 203 DISCUSIÓN Y CONCLUSIONES GENERALES…………………………………..211 REFERENCIAS………………………………………………………………………….233 ANEXOS………………………………………………………………………………….254 RESUMEN En el siguiente trabajo se describieron los estadios larvales de los digeneos parásitos presentes en los gasterópodos más abundantes de los intermareales de Puerto Madryn, Chubut (42°46'47"S, 65°00'06"O) y Puerto Deseado, Santa Cruz (47°45'15"S, 65°52'53"O): Buccinanops globulosus, Crepipatella dilatata, Fissurella radiosa, Kerguelenella lateralis, Nacella (Patinigera) magellanica, Pareuthria plumbea, Siphonaria lessonii, Tegula patagonica y Trophon geversianus. Siete de las nueve especies
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