Thesis AISLAMIENTO Y CARACTERIZACIÓN DE UNA BACTERIA HIDROCARBONOCLÁSTICA DE LA REGIÓN DE VALPARAÍSO Y SU APLICACIÓN EN BIORREMEDIACIÓN DE HIDROCARBUROS
Date
2018
Authors
HERNÁNDEZ GUERRERO, LISETTE ESTEPHANIE
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Abstract
La biorremediación es una estrategia para remover contaminantes desde suelos o aguas. Este proceso se lleva a cabo generalmente por bacterias degradadoras que fueron aisladas por sus capacidades catabólicas. La biorremediación de hidrocarburos ha sido estudiada como una tecnología efectiva y de bajo costo para la remoción de contaminantes. En el marco de esta memoria se planteó como objetivo aislar y caracterizar una nueva cepa degradadora y estudiar su posible aplicación en biorremediación de suelos contaminados con diésel. Se planteó como hipótesis: Desde un suelo proveniente de la Región de Valparaíso contaminado con diésel y sometido a un proceso de biorremediación se podrá aislar una cepa bacteriana degradadora de hidrocarburos. Los objetivos específicos fueron: (1) Aislar e identificar una cepa bacteriana hidrocarbonoclástica proveniente de un suelo contaminado con diésel y sometido a biorremediación; (2) Caracterizar la cepa bacteriana hidrocarbonoclástica aislada mediante técnicas microbiológicas y genéticas; (3) Evaluar las capacidades catabólicas de hidrocarburos del aislado bacteriano. El aislado se obtuvo desde muestras de microcosmos contaminados con diésel. La cepa aislada B7 se identificó como Achromobacter sp. que posee ¿ 99% de identidad con Achromobacter spanius LMG 5911 respecto al gen ARNr 16S. La cepa aislada B7 es capaz de crecer en medio rico en presencia hasta 6% de NaCl. La bacteria creció en hidrocarburos aromáticos como bifenilo y naftaleno. También se observó crecimiento en algunos alcanos como octano, hexano y hexadecano, y en diésel. La caracterización fenotípica y genotípica la cepa bacteriana B7 sugiere que se relaciona con una cepa de Achromobacter piechaudii. El estudio de la degradación de los componentes del diésel fue monitoreada mediante cromatografía gaseosa con detector por ionización por llama (GC-FID). El crecimiento se monitoreo midiendo unidades formadoras de colonias y turbidez. La degradación del sistema inoculado con la cepa B7 muestra alrededor de un 95 % de degradación de diésel considerando los hidrocarburos totales de petróleo (TPH), mientras que en el sistema abiótico se alcanzó un 50% de remoción de diésel. Además se observó una alta degradación de hidrocarburos del rango C15 – C21. Por sus propiedades degradadoras de hidrocarburos, la cepa B7 posee potencial para ser utilizada como parte de un consorcio o en forma individual para la biorremediación de hidrocarburos.
Bioremediation is a technology to remove contaminants from polluted sites. This process is usually carried out by degrading bacteria that were isolated based on their catabolic capabilities. The bioremediation of hydrocarbons has been studied as an effective and low-cost approach for the removal of contaminants. The objective of this thesis was to isolate and characterize a novel hydrocarbon-degrading strain and its application in bioremediation of soils contaminated with diesel. It was proposed as hypothesis: From a soil from the Valparaíso Region contaminated with diesel and subjected to a bioremediation process, a hydrocarbon-degrading bacterial strain can be isolated. The specific objectives were : (1) Isolate and identify a hydrocarbonoclastic bacterial strain from a soil contaminated with diesel and subjected to bioremediation; (2) Characterize the isolated hydrocarbonoclastic bacterial strain by microbiological and genetic techniques; (3) Evaluate the hydrocarbons catabolic capabilities of the bacterial isolate. The isolate was obtained from samples of a soil contaminated with diesel. The isolated strain B7 was identified as Achromobacter sp. showing ¿ 99% identity with Achromobacter spanius LMG 5911 with 16S rRNA gen. Strain B7 is able to grow in presence of up to 6% NaCl. The bacteria grew in aromatic hydrocarbons such as biphenyl and naphthalene. Growth was also observed on specific alkanes such as octane, hexane and hexadecane, and on diesel. The phenotypic and genotypic characterization of the bacterial strain B7 suggests that it belong to Achromobacter piechaudii. The degradation of diesel components was monitored by gas chromatography with flame ionization detector (GC-FID). Growth was monitored by measuring colony forming units and turbidity. The degradation of the system inoculated with the B7 strain shows ¿ 95% diesel degradation based on the total petroleum hydrocarbons (TPH), whereas in the abiotic system 50% diesel removal was achieved. High degradation of hydrocarbons of the C15 - C21 range was observed. Due to its hydrocarbon degrading capabilities, strain B7 has the potential to be used as part of a consortium or individually for the bioremediation of hydrocarbons.
Bioremediation is a technology to remove contaminants from polluted sites. This process is usually carried out by degrading bacteria that were isolated based on their catabolic capabilities. The bioremediation of hydrocarbons has been studied as an effective and low-cost approach for the removal of contaminants. The objective of this thesis was to isolate and characterize a novel hydrocarbon-degrading strain and its application in bioremediation of soils contaminated with diesel. It was proposed as hypothesis: From a soil from the Valparaíso Region contaminated with diesel and subjected to a bioremediation process, a hydrocarbon-degrading bacterial strain can be isolated. The specific objectives were : (1) Isolate and identify a hydrocarbonoclastic bacterial strain from a soil contaminated with diesel and subjected to bioremediation; (2) Characterize the isolated hydrocarbonoclastic bacterial strain by microbiological and genetic techniques; (3) Evaluate the hydrocarbons catabolic capabilities of the bacterial isolate. The isolate was obtained from samples of a soil contaminated with diesel. The isolated strain B7 was identified as Achromobacter sp. showing ¿ 99% identity with Achromobacter spanius LMG 5911 with 16S rRNA gen. Strain B7 is able to grow in presence of up to 6% NaCl. The bacteria grew in aromatic hydrocarbons such as biphenyl and naphthalene. Growth was also observed on specific alkanes such as octane, hexane and hexadecane, and on diesel. The phenotypic and genotypic characterization of the bacterial strain B7 suggests that it belong to Achromobacter piechaudii. The degradation of diesel components was monitored by gas chromatography with flame ionization detector (GC-FID). Growth was monitored by measuring colony forming units and turbidity. The degradation of the system inoculated with the B7 strain shows ¿ 95% diesel degradation based on the total petroleum hydrocarbons (TPH), whereas in the abiotic system 50% diesel removal was achieved. High degradation of hydrocarbons of the C15 - C21 range was observed. Due to its hydrocarbon degrading capabilities, strain B7 has the potential to be used as part of a consortium or individually for the bioremediation of hydrocarbons.
Description
Catalogado desde la version PDF de la tesis.
Keywords
BACTERIA HIDROCARBONOCLASTICA , BIORREMEDIACION , MICROBIOLOGIA
