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Haemocystidium Spp., a Species Complex Infecting Ancient Aquatic

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Haemocystidium Spp., a Species Complex Infecting Ancient Aquatic IDENTIFICACIÓN DE HEMOPARÁSITOS PRESENTES EN LA HERPETOFAUNA DE DIFERENTES DEPARTAMENTOS DE COLOMBIA. Leydy Paola González Camacho Universidad Nacional de Colombia Facultad de ciencias, Instituto de Biotecnología IBUN Bogotá, Colombia 2019 IDENTIFICACIÓN DE HEMOPARÁSITOS PRESENTES EN LA HERPETOFAUNA DE DIFERENTES DEPARTAMENTOS DE COLOMBIA. Leydy Paola González Camacho Tesis o trabajo de investigación presentada(o) como requisito parcial para optar al título de: Magister en Microbiología. Director (a): Ph.D MSc Nubia Estela Matta Camacho Codirector (a): Ph.D MSc Mario Vargas-Ramírez Línea de Investigación: Biología molecular de agentes infecciosos Grupo de Investigación: Caracterización inmunológica y genética Universidad Nacional de Colombia Facultad de ciencias, Instituto de biotecnología (IBUN) Bogotá, Colombia 2019 IV IDENTIFICACIÓN DE HEMOPARÁSITOS PRESENTES EN LA HERPETOFAUNA DE DIFERENTES DEPARTAMENTOS DE COLOMBIA. A mis padres, A mi familia, A mi hijo, inspiración en mi vida Agradecimientos Quiero agradecer especialmente a mis padres por su contribución en tiempo y recursos, así como su apoyo incondicional para la culminación de este proyecto. A mi hijo, Santiago Suárez, quien desde que llego a mi vida es mi mayor inspiración, y con quien hemos demostrado que todo lo podemos lograr; a Juan Suárez, quien me apoya, acompaña y no me ha dejado desfallecer, en este logro. A la Universidad Nacional de Colombia, departamento de biología y el posgrado en microbiología, por permitirme formarme profesionalmente; a Socorro Prieto, por su apoyo incondicional. Doy agradecimiento especial a mis tutores, la profesora Nubia Estela Matta y el profesor Mario Vargas-Ramírez, por el apoyo en el desarrollo de esta investigación, por su consejo y ayuda significativa con esta investigación. A la profesora Nubia Estela Matta, un agradecimiento especial por darme la oportunidad de pertenecer al grupo de investigación GERPH. Agradecimiento especial a todos mis compañeros del grupo de estudio “GERPH: Grupo de estudio de relación parásito–hospedero”, por su apoyo, especialmente a Germán Gutiérrez y Carolina Vargas, por su apoyo y comprensión a lo largo del desarrollo de esta investigación. A la profesora Martha Calderon, de la Universidad Nacional de Colombia, quien a través de las salidas de campo de la asignatura “Taxonomia animal”, el profesor Oscar Rodríguez de la Fundación Universitaria-Unitrópico y su grupo de investigación, a Rafael Gutiérrez y a Andrés Jiménez, quienes recolectaron algunas de las muestras analizadas; sin sus esfuerzos en campo el desarrollo de esta investigación no hubiera sido posible. VI IDENTIFICACIÓN DE HEMOPARÁSITOS PRESENTES EN LA HERPETOFAUNA DE DIFERENTES DEPARTAMENTOS DE COLOMBIA. Finalmente, quiero agradecer el apoyo financiero del proyecto Colciencias Número 018- 2018 “FP44842-018-2018” “Hemoparásitos asociados a la herpetofauna colombiana: Aspectos relevantes para su conservación”. Código 111877657872. Resumen y Abstract VII Resumen Colombia es un país altamente diverso en flora y fauna, siendo el 4to país con mayor biodiversidad de reptiles en el mundo. Sin embargo, los estudios relacionados con parásitos sanguíneos que infectan animales en vida silvestre son limitados. Apicomplexa es un phylum de protozoarios que agrupa parásitos obligados, como los hemoparásitos que infectan mamíferos, aves y reptiles. Este estudio evaluó la diversidad de hemoparásitos presentes en reptiles de diferentes departamentos, utilizando la determinación morfológica e información del marcador molecular citocromo b, para los géneros Plasmodium y Haemocystidum, y del fragmento de 18S rRNA, para el género Haemogregarinas. Se analizaron 225 individuos, encontrando 148 infectados con Haemogregarinas spp, cuatro con Plasmodium spp., y tres con Haemocystidium spp. Para Plasmodium kentropyxi y Plasmodium carmelinoi, encontrados en Cnemidophorus cf. gramivagus y Ameiva ameiva, respectivamente, se reportó por primera vez linajes de cytb que puden ser usados como BarCode (Capítulo 1). Se reporta la presencia de Haemocystidium sp en Podocnemis vogli (Capítulo 2). Se identificaron 14 secuencias de 18S rRNA asociados a dos morfotipos de Haemogregarinas spp. en Podocnemis vogli, y una secuencia asociada al único morfotipo en Podocnemis unifilis (Capítulo 3). El marcador citocromo b es útil para usarse como BarCode para identificar especies de haemosporidos; en el caso de las Haemogregarinas, la información obtenida con el marcador 18S rRNA, debe complementarse, por lo que sugiere la búsqueda de nuevos marcadores, que contribuyan a mejorar la resolución de las relaciones filogenéticas y la diferenciación entre e intra especies del suborden Adeleorina. Palabras clave: Tortugas, Apicomplexa, citocromo b, 18S rRNA. VII IDENTIFICACIÓN DE HEMOPARÁSITOS PRESENTES EN LA HERPETOFAUNA DE DIFERENTES I DEPARTAMENTOS DE COLOMBIA. Abstract Colombia is a highly diverse country in flora and fauna, being the fourth country with the greatest reptile biodiversity in the world. Nonetheless, the studies related to blood parasites that infect wildlife animals are limited. Apicomplexa is a protozoan’s phylum that groups obligate parasites, comprising hemoparasites infecting mammals, birds, and reptiles. In this study, the diversity of hemoparasites present in reptiles of different departments were evaluated by using morphological determination and molecular information of Cytochrome b molecular marker for Plasmodium and Haemocystidium genera, and a fragment of 18S rRNA for the Haemogregarina genus. 225 individuals were analyzed, 147 of them were positive for Haemogregarinas spp. infection, four for Plasmodium spp. and three for Haemocystidium spp. For Plasmodium kentropyxi y Plasmodium carmelinoi, found in Cnemidophorus cf. gramivagus y Ameiva ameiva, respectively, cytb linages that could be used as BarCode were reported for the first time (Chapter one). The Haemocystidium sp. was reported in Podocnemis vogli (Chapter two). 14 sequences of 18S rRNA were identified and associated with two morphotypes of Haemogregarinas spp. in Podocnemis vogli and one sequence was associated with a unique morphotype in Podocnemis unifilis (Chapter 3). Cytochrome b is a useful molecular marker as BarCode for haemosporidian species identification; the information obtained with 18S rRNA molecular marker was not sufficient for species identification for Haemogregarinas, therefore, we suggest to seek new markers that contribute to the resolution of phylogenetic relationships and the differentiation between and within species of Adelorina suborder. Keywords: Turtle, tortoise, Apicomplexa, Cytochrome b, 18S rRNA Contenido IX Contenido Lista de figuras .................................................................................. XII Lista de tablas ................................................................................... XIV Objetivos XV Objetivos específicos ..................................................................................................... XV Introducción ........................................................................................17 Referencias ..................................................................................................................... 20 Revisión de literatura ...................................................................................................... 24 Clasificación taxonómica................................................................................................. 24 Orden Haemosporidia (Haemosporina) .......................................................................... 26 Familia Plasmodiidae. ..................................................................................................... 27 Plasmodium spp. ............................................................................................................. 27 Familia Garnidae. ............................................................................................................ 28 Familia Haemoproteidae. ................................................................................................ 28 Ciclo de vida. ................................................................................................................... 29 Análisis filogenético. ........................................................................................................ 30 Suborden Adeleorina ...................................................................................................... 31 Familia Haemogregarinidae. ........................................................................................... 31 Familia Hepatozoidae ..................................................................................................... 31 Familia Karyolysidae: ...................................................................................................... 31 Familia Dactylosomatoidae. ............................................................................................ 32 Ciclo de vida .................................................................................................................... 32 Marcadores moleculares usados para en el estudio del suborden Adeleorina en herpetos ........................................................................................................................... 33 Referencias ..................................................................................................................... 34 1. Capítulo 1. Plasmodium parasites in reptiles from the Colombia Orinoco-Amazon
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