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Master's Dissertation
DOI
https://doi.org/10.11606/D.95.2023.tde-09102023-164409
Document
Author
Full name
Vinicius Sousa Flores
E-mail
Institute/School/College
Knowledge Area
Date of Defense
Published
São Paulo, 2023
Supervisor
Committee
Silva, Aline Maria da (President)
Ferreira, Henrique
Souza, Robson Francisco de
Varani, Alessandro de Mello
Title in Portuguese
Recuperação e caracterização de genomas de bacteriófagos a partir de metagenomas de compostagem
Keywords in Portuguese
Compostagem
Fagos
Genomas recuperados de metagenomas
Metagenoma
Abstract in Portuguese
Bacteriófagos ou fagos são vírus que infectam bactérias. Esses vírus causam grande impacto nas comunidades bacterianas dos mais diversos ambientes, sendo também centrais para a regulação de ciclos biogeoquímicos dos ambientes em que se encontram. Por muito tempo a sua diversidade e papel ecológico foi subestimada. O surgimento do sequenciamento de alto desempenho acompanhado do aprimoramento das técnicas de metagenômica permitiram um maior aprofundamento no estudo da diversidade e ecologia desses vírus. A recuperação de genomas montados a partir de metagenomas (ou Metagenome-Assembled Genomes - MAGs) permite a inferência do possível impacto que os genomas recuperados exercem sobre o ambiente. Nos últimos anos, compêndios de genomas de fagos têm sido construídos a partir de metagenomas de variados ambientes e suas implicações ecológicas têm sido exploradas. Entretanto, pouco ainda se sabe sobre o componente viral na compostagem, um ambiente altamente dinâmico em que as comunidades de microrganismos são definidas e reguladas pelo tipo de material compostado, disponibilidade de oxigênio, umidade e temperatura. O objetivo deste trabalho foi a construção de um catálogo de genomas de fagos recuperados de metagenomas de amostras de compostagem do Parque Zoológico de São Paulo. Para tal, realizamos a reconstrução de MAGs virais (vMAGs) com foco em fagos da classe Caudoviricetes. Foram recuperados 401 vMAGs de metagenomas de amostras coletadas em 6 composteiras distintas. A média de tamanho destes vMAGs foi 39 kbp, variando entre 10 kbp a 600 kbp. Um vMAG foi classificado como tendo seu genoma completo e 35 vMAGs foram classificados com alto grau de completude. Cerca de 50% dos vMAGs foram atribuídos a fagos com estilo de vida virulento, porém entre os vMAGs de maior completude o estilo de vida temperado foi predominante. A análise de clusterização apontou 77% dos clusters formados integralmente por vMAGs recuperados da compostagem, sugerindo que boa parte dos genomas recuperados pertencem a novos gêneros. Genes Metabólicos Auxiliares (Auxiliary Metabolic Genes AMGs) foram encontrados em 63 vMAGs, sendo ~6% de AMGs ligados ao metabolismo de carboidratos encontrados em plantas. Quinze vMAGs com AMGs foram ligados a MAGs bacterianos previamente recuperados e que possuem um papel central no metabolismo de carboidratos complexos. A análise do perfil de abundância dos vMAGs recuperados de duas das composteiras analisadas revelam um aumento gradual na abundância de fagos ao longo dos dias de compostagem.
Title in English
Recovery and characterization of bacteriophage genomes from composting metagenomes
Keywords in English
Composting
Metagenome
Metagenome-assembled genomes
Phage
Abstract in English
Bacteriophages or phages are viruses that infect bacteria. These viruses have a major impact on bacterial communities in a wide range of environments and are also central to the regulation of biogeochemical cycles in the environments in which they are found. For a long time, their diversity and ecological role were underestimated. The emergence of high-throughput sequencing, and the improvement of metagenomics techniques, have allowed a deeper study of the diversity and ecology of these viruses. The recovery of Metagenome-Assembled Genomes (MAGs) allows inferring the possible impact that the recovered genomes have on the environment. In recent years, compendia of phage genomes have been constructed from metagenomes from varied environments, and their ecological implications have been explored. However, little is known about the viral component in composting, a highly dynamic environment in which communities of microorganisms are defined and regulated by the composted material type, oxygen availability, moisture, and temperature. The aim of this work was the construction of a catalog of phage genomes retrieved from metagenomes of composting samples of the Sao Paulo Zoo Park. For that, we performed reconstruction of viral MAGs (vMAGs), focusing on phages of the class Caudoviricetes. We recovered 401 vMAGs from metagenomes of samples collected in 6 distinct composting piles. The vMAGs average size was 39 kbp ranging from 10 kbp to 600 kbp. One vMAG was classified as having its genome complete, and 35 vMAGs were classified with high degree of completeness. About 50% of the vMAGs were assigned to phages with virulent lifestyle, but among the vMAGs with the highest completeness, the temperate lifestyle was predominant. Clustering analysis indicated that 77% of the clusters were formed entirely by vMAGs recovered from composting, suggesting that most of the recovered genomes belong to new genera. Auxiliary Metabolic Genes (AMGs) were found in 63 vMAGs, with about 6% of AMGs linked to carbohydrate metabolism found in plants. Fifteen vMAGs with AMGs were linked to previously recovered bacterial MAGs that have a central role in complex carbohydrate metabolism. Analysis of the abundance profile of the vMAGs recovered from two of the analyzed composting piles reveals a gradual increase in phage abundance over the composting days.
 
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Publishing Date
2023-10-10
 
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