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Abatimiento De La Toxicidad De Compuestos Arsenicales Presentes En Suelos Agrícolas, a Través De Un Sistema De Tratamiento Biológico Aeróbico Secuencial

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Abatimiento De La Toxicidad De Compuestos Arsenicales Presentes En Suelos Agrícolas, a Través De Un Sistema De Tratamiento Biológico Aeróbico Secuencial Universidad de Concepción Dirección de Postgrado Facultad de Ciencias Biológicas Programa de Magíster en Ciencias con Mención Microbiología Abatimiento de la toxicidad de compuestos arsenicales presentes en suelos agrícolas, a través de un sistema de tratamiento biológico aeróbico secuencial Tesis para optar al grado de Magíster en Ciencias con Mención Microbiología JAVIERA FRANCISCA RAVANAL PINEDA CONCEPCIÓN-CHILE 2018 Profesor Guía: Víctor Campos Araneda Dpto. de Microbiología Facultad de Ciencias Biológicas Universidad de Concepción ©2018, JAVIERA FRANCISCA RAVANAL PINEDA Se autoriza la reproducción total o parcial, con fines académicos, por cualquier medio o procedimiento, incluyendo la cita bibliográfica del documento. Esta tesis ha sido realizada en el Departamento de Microbiología de la Facultad de Ciencias Biológicas, Universidad de Concepción. Profesor Guía ______________________________ Dr. Víctor Campos Araneda Facultad de Ciencias Biológicas Universidad de Concepción Comisión Evaluadora: ______________________________ Dr. Carlos Smith Facultad de Ciencias Biológicas Universidad de Concepción ______________________________ Dr. Miguel Martínez Poblete Facultad de Ciencias Biológicas Universidad de Concepción ______________________________ Dr. Víctor Hernández Facultad de Ciencias Biológicas Universidad de Concepción Director de Programa ______________________________ Dr. Víctor Campos Araneda Facultad de Ciencias Biológicas Universidad de Concepción “El techo de cristal en ciencia se ha movido, pero no se ha roto” Mª Ángeles Durán A mi familia, en especial a mis sobrinas, a quienes espero enseñarles que “No hay barrera, cerradura, ni cerrojo que puedas imponer a la libertad de mi mente” Virginia Woolf AGRADECIMIENTOS Desde siempre, transitar por el camino de un postgrado ha sido un enorme desafío personal, profesional y académico. Para mí, el desarrollo de este magíster ha sido el período de mi vida de mayor aprendizaje y crecimiento personal. Viví los momentos más difíciles y también lo más felices. Culminar este periodo con cuentas positivas, ha sido gracias a que en este camino ha habido personas que me enseñaron, me apoyaron, me tendieron su mano y me brindaron su amistad. En primer lugar, agradezco al Dr. Víctor Campos, tutor de esta tesis, por darme la oportunidad de continuar trabajando en las investigaciones desarrolladas en el Laboratorio de Microbiología Ambiental durante la ejecución de mi postgrado. Por su apoyo, guía y generosidad. A la Dra. María Angélica Mondaca, tutora de mi tesis de pregrado, quien me dio la oportunidad de integrar el equipo de investigación del Laboratorio de Microbiología Ambiental. Por su compromiso en el desarrollo de mi trabajo, por su paciencia, preocupación y, especialmente, por sus consejos y apoyo cuándo postulé por primera vez al programa de magíster y decidí esperar un año para postular nuevamente. A la Dra. Carla León, por brindarme su apoyo y amistad en los momentos difíciles, en los momentos confusos y en los momentos felices. Por su apoyo científico-académico en el desarrollo de mi trabajo, por su gran disposición y colaboración. A quienes fueron y son mis compañeros y amigos del Laboratorio de Microbiología Ambiental. En especial a Luis Pereira, por su cariño, su interés constante en colaborar y discutir ideas científicas y políticas. A Cristian Valenzuela, por las tantas conversaciones y por tener siempre una actitud agradable. A Benner Giacomozzi, por ser mi amigo durante su paso por el laboratorio y enseñarme lo que sé sobre curvas de crecimiento, calidad de agua v y rigurosidad experimental. A Juan Pablo Cuevas por su simpatía y compañía durante las interminables horas de trabajo. A Verónica Albornoz, por su carisma, ayuda y disposición. A mis compañeros de magíster por los gratos momentos vividos a pesar del cansancio. Especialmente a Cristian Tapia, Romina Carvajal y Javiera Llanos. A las funcionarias y funcionarios de la Facultad de Ciencias Biológicas y el Departamento de Microbiología. Por su fundamental trabajo, su cordialidad y su trato siempre cariñoso. A mis Bioamigos, me siento inmensamente agradecida por su apoyo, empatía y presencia en los momentos agotadores. A mis amigos, especialmente a Camila, Pedro y Daniela. Gracias por escucharme, quererme, retarme, apoyarme, reír y llorar conmigo. A mis abuelas(os), tías(os) y primas(os), por siempre estar pendientes e interesados en lo que estaba realizando. Agradezco a Juan, especialmente por ser mi apoyo emocional durante el periodo más difícil del desarrollo de este postgrado. Por compartir conmigo su paciencia y su amor. Por ser mi amigo y compañero en igualdad. A mis sobrinas, por enseñarme que se puede amar a alguien por el simple hecho de existir. Por enseñarme que las niñas y niños deben ser tratados como personas independientes y por motivarme a aprender más sobre educación y desarrollo cognitivo. A mis padres y a mi hermana, por su amor y apoyo incondicional. Por enseñarme a ser independiente, fuerte y que lo más importante es la felicidad. Por creer en mí y darme la confianza para soñar. Mi cimiento como persona son ustedes y son a quienes más amo. vi TABLA DE CONTENIDO AGRADECIMIENTOS ………………………………………….………………….. v ÍNDICE DE TABLAS .……………….............…………………….…………….. xii ÍNDICE DE FIGURAS …………......………………………………….……….. xviii RESUMEN …………...…………………………………………………….…….. xxii ABSTRACT ……………………………………………………………………… xxv I. INTRODUCCIÓN ......................................................................................... 1 1. Arsénico orgánico e inorgánico .......................................................... 4 2. Introducción de arsénico al ambiente por la industria avícola ......... 6 3. Introducción de arsénico al ambiente por la agricultura ................. 12 4. Arsénico en suelos de cultivo ........................................................... 17 5. Ciclo Biogeoquímico del arsénico .................................................... 20 6. Transformación biológica de compuestos organoarsenicales ....... 25 7. Transformación bacteriana de arsénico inorgánico ........................ 28 8.Precipitación de carbonato de calcio inducida por microorganismos (MICP) ...................................................................................................... 34 9.Biomineralización acoplada a la oxidación de arsenito e interacción calcita-arsénico ...................................................................................... 35 10. Tratamiento biológico de aguas contaminadas con arsénico ...... 38 II. HIPÓTESIS Y OBJETIVOS…………………………………………………...43 11. Hipótesis ........................................................................................... 43 12. Objetivos ........................................................................................... 45 vii III. METODOLOGÍA ..................................................................................... 46 13.Caracterización del consorcio bacteriano crecido en presencia de compuestos organoarsenicales ............................................................ 46 13.1 Consorcio Bacteriano ................................................................... 46 13.2 Condiciones de cultivo del consorcio bacteriano .......................... 48 13.3 Cinética de crecimiento del consorcio bacteriano ......................... 49 13.4 Reducción de compuestos organoarsenicales ............................. 49 14. Caracterización de la actividad ureolítica-calcificante y arsenito- oxidante de Pseudomonas arsenicoxydans ........................................ 50 14.1 Pseudomonas arsenicoxydans ..................................................... 50 14.2 Actividad ureolítica de P. arsenicoxydans .................................... 51 14.3Precipitación de calcita microbiológicamente inducida (MICP) por P. arsenicoxydans .................................................................................... 52 14.4 Condiciones de cultivo de P. arsenicoxydans ............................... 53 14.5 Cultivos de P. arsenicoxydans en estado planctónico .................. 53 14.6 Cultivos de P. arsenicoxydans en estado sésil ............................. 55 14.7 Cinética de crecimiento de P. arsenicoxydans ............................. 56 14.8 Capacidad de remover arsénico de P. arsenicoxydans ................ 57 14.9 Biopelícula y cristales de calcita formados durante la MICP ......... 57 14.10 Cultivo de P. arsenicoxydans en presencia de efluente abatido por consorcio bacteriano ...................................................................... 58 15. Sistema de tratamiento biológico aeróbico secuencial ................. 59 viii 15.1 Cinéticas de crecimiento del consorcio bacteriano y P. arsenicoxydans .................................................................................... 61 15.2 Reducción de compuestos organoarsenicales y oxidación/co- precipitación de arsénico ...................................................................... 61 15.3 Análisis de la biopelícula del biorreactor semi-continuo ................ 62 15.4 Evaluación de la toxicidad del efluente final del sistema de tratamiento biológico aeróbico secuencial ............................................ 63 16.Determinación analítica de cinéticas de aparición y oxidación de arsénico a través de HPLC-HG-AAS ..................................................... 64 17. Análisis estadísticos y parámetros cinéticos ................................. 65 IV. RESULTADOS .......................................................................................
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