Perfil das comunidades de Archaea em sistemas anaeróbios de tratamento de efluentes domésticos utilizando a eletroforese em gel de gradiente desnaturante (DGGE)

• Autor(a) principal: Bettio, Guilherme Pianaro
• Data de Publicação: 2012
• Tipo de documento: Dissertação
• Idioma: por
• Título da fonte: Repositório do Centro Universitário Braz Cubas
• Texto Completo: https://repositorio.cruzeirodosul.edu.br/handle/123456789/2555
• topic: DGGE; PCR; Archaea; Tratamento anaeróbio; Biogás; Extração de DNA; CNPQ::CIENCIAS BIOLOGICAS::BIOLOGIA GERAL

In the present work sludge samples were collected from two domestic wastewater treatment stations of Sanepar, Curitiba-PR. One sample was taken from the anaerobic lagoon (LA) and the other from the fluidized bed anaerobic reactor (RALF). Eight procedures were employed for DNA extraction from the communities present in the collected samples, and two of them (extraction procedures 1 and 2) were effective in obtaining a quality product. After obtaining DNA of quality, for both samples LA and RALF, amplifications of target fragments encoding the 16S rDNA region of Archaea were made. Amplifications were performed by PCR using the primers PRA46f and PREA1100r, followed by Nested PCR using the primers PARCH340f and PARCH519r. Amplification conditions were modified many times in order to achieve higher quality and specificity of the final product. After amplification of the fragments of around 180 bp by Nested PCR, these were purified and submitted to sequencing. The fragments were also submitted to denaturing gradient gel electrophoresis (DGGE), a technique that generated profiles of the communities present in both systems. In this way, the comparison of these profiles was possible through the principal component analysis (PCA), based on the Bray-Curtis method, allowing to verify the difference in the profile obtained for the samples RALF and LA, that indicates the difference in composition of the communities present in both systems. The similarity degree was of 65% for the communities existent in the different environments.