Análise metagenômica e potencial biotecnológico de microrganismos de solo e água de uma área agrícola com adubação orgânica
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Data
2016-12-16
Autores
Meneghine, Aylan Kener [UNESP]
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Universidade Estadual Paulista (Unesp)
Resumo
O composto orgânico produzido a partir de carcaças, resíduos animais e vegetais é uma alternativa viável para a substituição total ou parcial dos fertilizantes minerais utilizados na atualidade. No processo de compostagem participam diferentes populações microbianas, e com isso o composto torna-se um sistema rico para utilização como fertilizante no solo, complementando assim as necessidades nutricionais e microbianas do meio ambiente. Entretanto, há poucos trabalhos envolvendo análise da diversidade bacteriana em solos sob uso de composto orgânico feito a partir de carcaças, e também pouco se conhece sobre o impacto ambiental do uso agrícola de composto orgânico na qualidade da água. Existe também a questão se há influência da água utilizada para irrigação na qualidade do solo. O objetivo central desse trabalho foi analisar a diversidade bacteriana e perfil funcional de um solo de horta e da água de um córrego utilizada para irrigação. E como objetivo secundário, através do isolamento de bactérias da água verificar o potencial biotecnológico de produção e uso de exopolissacarídeo como bioemulsificante de óleo e hidrocarbonetos. As amostras de solo e água utilizadas nesse trabalho foram coletadas na área do departamento rural da Fundação Parque Zoológico de São Paulo, em setembro de 2014. Todo material coletado foi transportado até o Laboratório de Bioquímica de Micro-organismos e de Plantas, onde realizou-se a extração de DNA total e sequenciamento através de tecnologia Ion Proton (Life Technologies), isolamento bacteriano, produção e aplicação de exopolissacarídeo como bioemulsificante. Ao analisar o DNA metagenômico observou-se que tanto na água quanto no solo há uma abundância de bactérias encontradas normalmente em áreas agrícolas sob influência de composto orgânico feito a partir de outras fontes. Verificou-se também, através da análise dos genes relacionados com os ciclos biogeoquímicos, abundância de genes do processo de desnitrificação na amostra de solo. Além de se observar que na análise da estrutura taxonômica dos ciclos biogeoquímicos, não há bactérias em comum entre as amostras de solo e água com do composto orgânico. Porém ao avaliar o perfil funcional total da água, solo e outros dez metagenomas, através de uma análise de agrupamento hierárquico, notou-se que a água causa maior influência no solo do que o próprio composto orgânico. Com relação ao isolamento bacteriano, foi possível isolar a bactéria Sphingomonas sp., sendo que o isolado identificado demonstrou produzir grande quantidade de exopolissacarídeo (EPS). O EPS formou emulsões estáveis após 24 horas e 168 horas. Além disso o EPS foi um melhor agente emulsificante para hexano e querosene dentre todos os óleos analisados. Os resultados mostraram contribuição significativa com relação ao potencial do EPS como agente bioemulsificante que aumentaria a degradação de hidrocarbonetos e efeitos de emulsificação em biotecnologia ambiental.
The organic compost produced from carcasses, animal and vegetable waste is a viable alternative to full or partial replacement of mineral fertilizers used nowadays. In the composting process there are involved different microbial populations, and the compost becomes a rich system for use as a fertilizer in the soil, thereby supplementing the nutritional and microbial requirements of the medium. However, there are few studies involving the analysis of bacterial diversity in soil under use of organic compost made from carcasses, and also little is known about the environmental impact of agricultural use of organic compost in water quality. Furthermore, there is also the question of whether there is influence of the water used for irrigation on soil quality. Because of these questions, the central objective of this study was to analyze the bacterial diversity and functional profile of a soil from vegetable garden and freshwater used for irrigation from a local stream. As a secondary objective, we aimed to verify the production and biotechnology potential of a bacterial exopolysaccharide as oil and hydrocarbons bioemulsifier. Soil and freshwater samples used in this study were collected at rural department of the Zoo Foundation Park of São Paulo, in September 2014. All material collected was transported to the Laboratório de Bioquímica de Micro-organismos e de Plantas where we proceeded with the total DNA extraction experiments, sequencing through Ion Proton technology (Life Technologies), bacterial isolation, production and application of exopolysaccharide as bioemulsifier. By analyzing the metagenomic DNA it was observed that both freshwater and soil were plenty of bacterial communities normally found in agricultural areas under influence of organic amendments. Through the analysis of genes related to biogeochemical cycles, it was found abundance of genes about denitrification process in the soil sample. Also it can be noted by the analysis of the taxonomic structure of biogeochemical cycles, there was no bacteria shared between soil and freshwater samples with the organic compost. But when evaluating the functional profile of freshwater, soil and other ten metagenomes, we observed that freshwater causes greater influence on the soil than the organic compost. Regarding the bacterial isolation, it was possible to identify the bacteria Sphingomonas sp., and it has been shown to produce large amount of exopolysaccharide (EPS). The EPS has shown stability in its emulsions after 24 hours and 168 hours. Additionally, it’s a better emulsifying agent for hexane and kerosene among other oils analysed. Furthermore, our findings promoted significant contribution by showing EPS potential as a bioemulsifier agent that would enhance hydrocarbon degradation and emulsification effects in environmental biotechnology.
The organic compost produced from carcasses, animal and vegetable waste is a viable alternative to full or partial replacement of mineral fertilizers used nowadays. In the composting process there are involved different microbial populations, and the compost becomes a rich system for use as a fertilizer in the soil, thereby supplementing the nutritional and microbial requirements of the medium. However, there are few studies involving the analysis of bacterial diversity in soil under use of organic compost made from carcasses, and also little is known about the environmental impact of agricultural use of organic compost in water quality. Furthermore, there is also the question of whether there is influence of the water used for irrigation on soil quality. Because of these questions, the central objective of this study was to analyze the bacterial diversity and functional profile of a soil from vegetable garden and freshwater used for irrigation from a local stream. As a secondary objective, we aimed to verify the production and biotechnology potential of a bacterial exopolysaccharide as oil and hydrocarbons bioemulsifier. Soil and freshwater samples used in this study were collected at rural department of the Zoo Foundation Park of São Paulo, in September 2014. All material collected was transported to the Laboratório de Bioquímica de Micro-organismos e de Plantas where we proceeded with the total DNA extraction experiments, sequencing through Ion Proton technology (Life Technologies), bacterial isolation, production and application of exopolysaccharide as bioemulsifier. By analyzing the metagenomic DNA it was observed that both freshwater and soil were plenty of bacterial communities normally found in agricultural areas under influence of organic amendments. Through the analysis of genes related to biogeochemical cycles, it was found abundance of genes about denitrification process in the soil sample. Also it can be noted by the analysis of the taxonomic structure of biogeochemical cycles, there was no bacteria shared between soil and freshwater samples with the organic compost. But when evaluating the functional profile of freshwater, soil and other ten metagenomes, we observed that freshwater causes greater influence on the soil than the organic compost. Regarding the bacterial isolation, it was possible to identify the bacteria Sphingomonas sp., and it has been shown to produce large amount of exopolysaccharide (EPS). The EPS has shown stability in its emulsions after 24 hours and 168 hours. Additionally, it’s a better emulsifying agent for hexane and kerosene among other oils analysed. Furthermore, our findings promoted significant contribution by showing EPS potential as a bioemulsifier agent that would enhance hydrocarbon degradation and emulsification effects in environmental biotechnology.
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Palavras-chave
Bioemulsificante, Ciclos biogeoquímicos, Exopolissacarídeo, Bioemulsifier, Biogeochemical cycles, Exopolysaccharide