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Tesis Doctoral Dpto. Inmunología, Microbiología y Parasitología Immunologia, Mikrobiologia eta Parasitologia Saila Tesis Doctoral Efecto del cambio climático en la costa vasca: presencia de bacterias del género Vibrio e influencia de los factores ambientales en su perdurabilidad. Elixabet Ogayar Sandoval Leioa 2021 (c) 2021 Elixabet Ogayar Sandoval Dpto. Inmunología, Microbiología y Parasitología Immunologia, Mikrobiologia eta Parasitologia Saila TESIS DOCTORAL Efecto del cambio climático en la costa vasca: presencia de bacterias del género Vibrio e influencia de los factores ambientales en su perdurabilidad. ELIXABET OGAYAR SANDOVAL Leioa, 2021 UNIVERSIDAD DEL PAIS VASCO EUSKAL HERRIKO UNIBERTSITATEA FACULTAD DE CIENCIA Y TECNOLOGÍA Departamento de Inmunología, Microbiología y Parasitología Efecto del cambio climático en la costa vasca: presencia de bacterias del género Vibrio e influencia de los factores ambientales en su perdurabilidad. ELIXABET OGAYAR SANDOVAL Directoras: INÉS ARANA BASABE MAITE ORRUÑO BELTRÁN La presente Tesis Doctoral ha sido realizada gracias a la financiación recibida de los siguientes proyectos de investigación: Beca de Investigación Predoctoral UPV/EHU. Abril 2016 – Abril 2020. PPP 2017-3686 (UPV/EHU) Título del proyecto: Presencia de bacterias del género Vibrio en la costa vasca e influencia de los factores ambientales en su supervivencia Entidad financiadora: Universidad del País Vasco Duración: 2016 - 2018 Investigador responsable: Maite Orruño Los resultados obtenidos durante esta Tesis Doctoral han sido objeto de las siguientes publicaciones y contribuciones a congresos: Publicaciones: - Orruño M, Parada C, Ogayar E, Kaberdin VR, Arana I (2019). Effects of abiotic and biotic factors on Vibrio harveyi ATCC 14126T survival dynamics in seawater microcosms. Aquatic Microbial Ecology 83:109-118. DOI: 10.3354/ame01910. - Montánchez I, Ogayar E, Plágaro A.H, Esteve-Codina A, Gómez-Garrido J, Orruño M, Arana I, Kaberdin VR (2019). Analysis of Vibrio harveyi adaptation at elevated temperature reveals the putative mechanisms associated with its persistence and spread in the time of global warming. Scientific Reports 9:289. DOI:10.1038/s41598-018-36483-0. Contribuciones a congresos: - Ogayar E, Orruño M, Catediano R, Escobero A, Gallego B, González-Uriarte A, Ramos L, Kaberdin VR, Arana I. 2016. Efecto de la temperatura y la salinidad en cepas de Vibrio de origen ambiental y en la cepa de referencia V. harveyi ATCC 14126T. XI Congreso de Microbiología del Medio Acuático (XI-MMA’16).Oviedo, España. 20-22 julio 2016. - Parada C, Orruño M, Bravo Z, Ogayar E, Kaberdin VR, Arana I. 2016. Diferencias y semejanzas en las estrategias de supervivencia desarrolladas por Escherichia coli y Vibrio harveyi mantenidas bajo condiciones de estrés. XI Congreso de Microbiología del Medio Acuático (XI- MMA’16). Oviedo, España. 20-22 julio 2016. - Ogayar E, Orruño M, Montánchez I, Kaverdin VR, Arana I. 2016. Isolation of Vibrio species from the Bay of Biscay. XV International Symposium on Oceanography of the Bay of Biscay. Bilbao, España. 22-24 junio 2016. - Ogayar E, Orruño M, Arana I. 2016. Vibrio spp. en el litoral vizcaíno. V Jornadas de Investigación de la Facultad de Ciencia y Tecnología. Leioa, España. 6-7 abril 2016 - Bravo Z, Parada C, Ogayar E, Montánchez I, Ramos L, Catediano R, Gallego B, Escobero A, Orruño M, Kaberdin VR, Arana I. 2016. Bacterial resistance to stress. Survival strategies of Vibrio spp. V Jornadas de Investigación de la Facultad de Ciencia y Tecnología. Leioa, España. 6-7 abril 2016. - Orruño M, Ogayar E, Argos P, González-Uriarte A, Zaldibar B, Kaberdin VR, Arana I. 2015. Presencia de Vibrio spp. en el litoral vizcaíno. Resultados preliminares. XXV Congreso Nacional de Microbiología. Logroño, España. 7-10 julio 2015. - Orruño M, Parada C, Bravo Z, Ogayar E, Larrañaga I, Kaberdin VR, Arana I. 2017. Effects of long-term starvation on Vibrio harveyi membrane subproteome during its permanence in seawater. Congress of European Microbiologists. FEMS 2017. Valencia, España. 9-13 julio 2017. - Orruño M, Parada C, Ogayar E, Kaberdin V.R, Arana I. 2017. Effect of temperature, starvation and predation in the survival of Vibrio harveyi populations. HDID V (How dead is dead?). 5th Conference on Exploring the Edge of Bacterial Life. Vienna, Austria. 6-8 septiembre 2017. - Ogayar E, Orruño M, Arana I. 2018. Presence of Vibrio spp in the Basque coastal area and influence of environmental factors on their survival. VI Jornadas de Investigación de la Facultad de Ciencia y Tecnología. Leioa, España. 14-15 marzo 2018. - Orruño M, Kaberdin VR, Arana I, Ogayar E, Pascual A, Lomba A, González N. 2018. Bacterial resistance to stress. Vibrio spp. in the aquatic systems. VI Jornadas de Investigación de la Facultad de Ciencia y Tecnología. Leioa, España. 14-15 marzo 2018. - Ogayar E, Lomba A, Larrañaga I, Kaberdin VR, Arana I, Orruño M. 2018. Utilidad de la técnica CARD-FISH para estudiar la distribución de las bacterias del género Vibrio. XVII Reunión del Grupo Taxonomia, Filogenia y Biodiversidad y XII Reunión del Grupo de Microbiología del Medio Acuático. Sitges, España. 1-2 octubre 2018. - Orruño M, Ogayar E, Hernández A, Eguiraun H, Martinez I, Arana I, Kaberdin VR. 2018. Analysis of bacterial stress responses associated with climate change. MikrobioGune. 1st Basque Microbiology Meeting. Bilbao, España. 11 diciembre 2018. - Ogayar E, Ruiz T, Sainz I, Uriarte F, Ordoñez I, Orruño M, Arana I. 2019. Efecto del cambio climático en la costa Vasca: presencia de bacterias del género Vibrio e influencia de los factores ambientales en su perdurabilidad. Concurso de Divulgación Científica UPV/EHU. Bilbao, España. 11 abril 2019. - Kaberdin VR, Montánchez I, Ogayar E, Plágaro AH, Esteve-Codina A, Gómez-Garrido J, Orruño M, Arana I. 2019. Addressing the impact of temperature on Vibrio harveyi adaptation in the time of climate change. Symposium on Bacterial Genetics and Ecology (BAGECO). Lisboa, Portugal. 26-30 mayo 2019. - Orruño M, Ogayar E, Alonso-Sáez L, Kaberdin VR, Arana I. 2019. Seasonal distribution of Vibrio spp. populations in the coastal waters of the Bay of Biscay. 8th Congress of European Microbiologists. FEMS 2019. Glasgow, Escocia. 7-11 julio 2019. - Ogayar E, Ruiz T, Orruño M, Martínez I, Kaberdin VR, Arana I. 2019. Colonización de superficies bióticas por Vibrio harveyi. Hacia el diseño de un modelo animal-Vibrio. XXVII Congreso Nacional de Microbiología. Málaga, España. 2-5 julio 2019. Al hombre más inteligente que conozco, por el deseo de ser como él, hoy soy Doctora, mi padre. AGRADECIMIENTOS Mediante estas palabras me gustaría agradecer a todas las personas que han hecho posible la realización de esta tesis. En primer lugar a mis directoras Inés Arana y Maite Orruño, que confiaron en mi y me propusieron llevar a cabo este proyecto. Por enseñarme, pero sobre todo por darme la libertad para fallar y aprender por mi misma. Muchas veces nos preguntan que queremos ser de mayor, o cuales son nuestros sueños. Mi respuesta era “trabajar con bata de laboratorio”, gracias por hacerlo posible. Por supuesto, a todos mis compañeros y todos los alumnos internos que han pasado por el laboratorio. Agradezco toda vuestra ayuda, y no solo en el ámbito científico sino sobre todo en el ámbido personal. He sido muy feliz estos años en el laboratorio. A mi familia, que se han interesado por cada uno de los pasos que he dado, y ha celebrado cada uno de ellos como nadie. Sobre todo a mis padres, que siempre me han ayudado en todo y me han dado toda la confianza para lograr lo que me proponga. A todos mis amigos, que siempre han estado pendientes de mí, y me han ayudado a evadirme cuando más lo he necesitado, incluso desde la distancia. Gracias por llenarme el alma. A todos los que me habeis acompañado en este camino, hay un trocito de vosotros mismos en esta tesis (que sólo los más observadores sabrán encontrar). Mila esker! ÍNDICE ÍNDICE 3 ÍNDICE ______________________________________________________________________ 3 ÍNDICE DE FIGURAS _______________________________________________________ 7 ÍNDICE DE TABLAS _______________________________________________________ 13 INTRODUCCIÓN _____________________________________________________________ 18 1. Calentamiento global y Vibrio __________________________________________ 19 2. Problemática en la identificación de Vibrio spp. ____________________________ 21 3. Detección de Vibrio spp. en muestras ambientales __________________________ 23 4. Supervivencia de Vibrio spp. ___________________________________________ 24 OBJETIVOS _________________________________________________________________ 31 OBJETIVO 1: Distribución y dinámica espacio-temporal de Vibrio spp. en aguas costeras del golfo de Vizcaya ________________________________________________________ 32 MATERIALES Y MÉTODOS ____________________________________________________ 32 1. Adecuación de la técnica CARD-FISH (Catalyzed Reporter Deposition-Fluorescence In situ Hybridization) para la detección de Vibrio spp. ___________________________ 35 1.1. Elección de las sondas para la detección y enumeración de Vibrio spp. ______ 35 1.2. Procedimiento estandarizado _______________________________________ 38 1.3. Verificación de la idoneidad de las sondas _____________________________ 42 2. Enumeración de Vibrio spp. en la costa vasca a lo largo de un ciclo anual ________ 44 2.1. Área de muestreo ________________________________________________ 44 2.2. Procesamiento y análisis de las muestras ______________________________ 46 3. Aislamiento e identificación
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