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Estudio Del Crecimiento De Microcystis Aeruginosa Y De La Producción De Microcystina En Cultivo De Laboratorio

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Estudio Del Crecimiento De Microcystis Aeruginosa Y De La Producción De Microcystina En Cultivo De Laboratorio UNIVERSIDAD NACIONAL DE LA PLATA FACULTAD DE CIENCIAS EXACTAS DEPARTAMENTO DE CIENCIAS BIOLÓGICAS Trabajo de Tesis Doctoral ESTUDIO DEL CRECIMIENTO DE MICROCYSTIS AERUGINOSA Y DE LA PRODUCCIÓN DE MICROCYSTINA EN CULTIVO DE LABORATORIO Melina Celeste Crettaz Minaglia (Leda Giannuzzi / Darío Andrinolo) 2018 1 Dedicada a mi mamá. 2 Contenidos Resumen ........................................................................................................................................................................ 5 Introducción ................................................................................................................................................................... 9 Generalidades de las cianobacterias y cianotoxinas .................................................................................................. 9 Microcystis aeruginosa y microcistinas .................................................................................................................. 14 Factores de crecimiento de cianobacterias, en especial M. aeruginosa, y de producción de microcistinas ............ 19 Temperatura ........................................................................................................................................................ 19 Nutrientes ............................................................................................................................................................ 21 Irradiación ........................................................................................................................................................... 23 Modelos matemáticos .............................................................................................................................................. 26 Remoción de microcistinas en agua ........................................................................................................................ 31 Microorganismos con potencial para remover microcistinas .............................................................................. 33 Planteo del problema ................................................................................................................................................... 36 Hipótesis ...................................................................................................................................................................... 37 Hipótesis principales ............................................................................................................................................... 37 Objetivos...................................................................................................................................................................... 37 Objetivo general ...................................................................................................................................................... 37 Objetivos específicos............................................................................................................................................... 37 Materiales y métodos ................................................................................................................................................... 39 Cultivo de M. aeruginosa ........................................................................................................................................ 39 Aclimatación del cultivo ......................................................................................................................................... 40 Diseño experimental de la cinética de crecimiento y la producción de microcistina-LR ........................................ 41 Determinaciones en el estudio cinético de los cultivos ........................................................................................... 42 Determinación de la densidad óptica .................................................................................................................. 42 Recuento celular ................................................................................................................................................. 42 Determinación de clorofila-a .............................................................................................................................. 43 Determinación de microcistina-LR ..................................................................................................................... 44 Modelado matemático de los datos experimentales ................................................................................................ 45 Estimación de los parámetros cinéticos de la curva de crecimiento de M. aeruginosa ....................................... 45 Efecto de la temperatura sobre la tasa específica de crecimiento y la duración de la fase de latencia ................ 48 Modelado de la producción de clorofila-a .......................................................................................................... 51 Calculo del cuota (Qclo-a) .................................................................................................................................... 52 Modelado de la producción de microcistina-LR ................................................................................................. 52 Cálculo del cuota (Qclo-a y QMC) .......................................................................................................................... 53 Modelo superficie-respuesta ............................................................................................................................... 53 Toma de muestras ambientales para aislamiento del ensamble de microorganismos heterótrofos con potencial remoción de MC-LR ............................................................................................................................................... 54 3 Aclimatación y aislamiento de bacterias con potencial de remoción de microcistina-LR ...................................... 55 Extracción de microcistina [D-Leu1]- LR ........................................................................................................... 57 Ensayo de remoción de microcistina-LR ................................................................................................................ 58 Identificación........................................................................................................................................................... 59 Análisis estadístico .................................................................................................................................................. 60 Resultados y discusión ................................................................................................................................................. 60 Efecto de la temperatura de incubación sobre la cinética de crecimiento, la producción de clorofila-a y de microcistina-LR en un cultivo de M. aeruginosa .................................................................................................... 60 Efecto de la temperatura de incubación en la cinética de crecimiento de M. aeruginosa ................................... 60 Efecto de la temperatura de crecimiento de M. aeruginosa sobre la velocidad especifica de crecimiento (µ) y la fase de latencia (LPD) ......................................................................................................................................... 64 Efectos de la temperatura en la producción de clorofila-a .................................................................................. 68 Efectos de la temperatura sobre el valor de Qclo-a ............................................................................................... 71 Efectos de la temperatura en la producción de microcistinas en el cultivo de M. aeruginosa ............................ 71 Efectos de la temperatura sobre el parámetro QMC ............................................................................................. 75 Efectos de diferentes factores sobre el crecimiento de M. aeruginosa .................................................................... 77 Efectos de la temperatura, la irradiancia y la relación N:P sobre la cinética de crecimiento de M. aeruginosa . 77 Efectos de la temperatura, la irradiancia y la relación N:P sobre la producción de clorofila-a ........................... 91 Efectos de la temperatura, la irradiancia y la relación N:P sobre los valores de Qclo-a ...................................... 100 Efectos de la temperatura, la irradiancia y la relación N:P sobre la producción de microcistina-LR ................... 106 Efectos de la temperatura, la irradiancia y la relación N:P sobre QMC .............................................................. 116 Interacciones de la temperatura, la irradiación y la relación N:P sobre el crecimiento y la producción de clorofila- a y microcistinas .................................................................................................................................................... 122 Correlación entre los factores y los parámetros de crecimiento, la producción de clorofila-a y microcistina-LR 129 Vinculación de la velocidad de crecimiento y la producción de microcistina-LR ................................................ 129 Caracterización de la floración en el Río de La Plata ...........................................................................................
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