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Metabolic processes and Microbiome in a water supply reservoir during
cyanobacteria blooms using Next generation sequencing
Hoffmann, L.1,2; Cabral, B.C.A.1; Penha, L.1; Sales, E.B.1; Valentin de Souza, E.1,2;
Ürményi, T.P.1,2; Rondinelli, E.1,2; Azevedo, S.M.1,2; Silva, R.1,2
1
Instituto de Biofísica Carlos Chagas Filho, Universidade Federal do Rio de Janeiro,
Rio de Janeiro, Brazil;
2
Instituto Nacional para Pesquisa Translacional em Saúde e Ambiente na Região
Amazônica, Conselho Nacional de Desenvolvimento Científico e
Tecnológico/MCT, RJ
INTRODUCTION: Freshwater reservoirs are essential resource for human life. The
presence of toxins released by microbes, such as Cyanobacteria, may in large
quantities be prejudicial to humans and the ecosystem. Metagenomic studies
contribute to understand the dynamics and molecular basis of cyanobacteria blooms
and other microorganisms, leading to the identification of functional metabolic profiles
and the taxonomic classification of the microbiome. OBJECTIVES: To provide an
overview of the phylogenetic diversity of the bacterial community and relevant
metabolic activities in a water supply reservoir during blooms of cyanobacteria.
METHODS: Water samples (Samuel reservoir, State of Rondonia) were collected.
DNA was extracted by organic method. Next generation sequencing (NGS) using Ion
Torrent technology was done according the protocol recommendation. Three libraries
were constructed: 1) Shotgun library was performed using enzymatic shear protocol,
2) Ribosomal 16SrDNA using universal bacterial primers and 3) Ribosomal 16SrDNA
using cyanobacteria specific primers. Libraries were amplified using emulsion PCR
and were sequenced in a 314 chip for 16SrDNA barcoded library and a 314 and 316
chip for shotgun library. Sequences were submitted to Blastn and Blastx search to
find regions of similarity between biological sequences. Output files were analyzed
using MEGAN software. RESULTS: A total of 4,5 million reads for shotgun library and
650 thousand reads for both 16SrDNA libraries were obtained. Preliminary analysis of
NGS reads showed a high variability of species, especially for bacteria. The most
common Phyla were Proteobacteria and Firmicutes. KEGG profile showed great
amount of reads in metabolism pathway followed by environmental information
processes. DISCUSSION: Detailed analyses are being performed in order to
compare seasonal samples. Profile characterization of viroma and the identification of
uncultured microorganisms and potentially still unknown are important to evaluate the
dynamics of substitutions in the water reservoir microbiome and metabolic activities
relevant to understand the cyanobacteria blooms.
Key words: metagenomic, next generation sequencing, 16S ribosomal DNA,
bioinformatics, water reservoir
Financial support: INCT-INPeTAm/CNPq/MCT, edital PensaRio FAPERJ, and Visiting
Professor –Science without borders – CNPq.