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Illumina sequencing-based analysis of a microbial community enriched under anaerobic methane oxidation condition coupled to denitrification revealed coexistence of aerobic and anaerobic methanotrophs

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Abstract

Methane is produced in anaerobic environments, such as reactors used to treat wastewaters, and can be consumed by methanotrophs. The composition and structure of a microbial community enriched from anaerobic sewage sludge under methane-oxidation condition coupled to denitrification were investigated. Denaturing gradient gel electrophoresis (DGGE) analysis retrieved sequences of Methylocaldum and Chloroflexi. Deep sequencing analysis revealed a complex community that changed over time and was affected by methane concentration. Methylocaldum (8.2%), Methylosinus (2.3%), Methylomonas (0.02%), Methylacidiphilales (0.45%), Nitrospirales (0.18%), and Methanosarcinales (0.3%) were detected. Despite denitrifying conditions provided, Nitrospirales and Methanosarcinales, known to perform anaerobic methane oxidation coupled to denitrification (DAMO) process, were in very low abundance. Results demonstrated that aerobic and anaerobic methanotrophs coexisted in the reactor together with heterotrophic microorganisms, suggesting that a diverse microbial community was important to sustain methanotrophic activity. The methanogenic sludge was a good inoculum to enrich methanotrophs, and cultivation conditions play a selective role in determining community composition.

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Abbreviations

ANME:

Anaerobic methanotrophic archaea

DAMO:

Denitrifying anaerobic methane oxidation

DGGE:

Denaturing gradient gel electrophoresis

OTU:

Operational taxonomic unit

UASB:

Upflow anaerobic sludge blanket reactor

VTS:

Volatile total solids

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Acknowledgments

We wish to thank the Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES), Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq), and Fundação de Amparo a Pesquisa do Estado de Minas Gerais (FAPEMIG). We are also grateful to Dr. Emanuel F. Brandt for helping with the methane mass balance calculation.

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Author notes

  1. Luciene Alves Batista Siniscalchi

    Present address: Department of Engineering, Federal University of Lavras, Dr. Sylvio Menicucci Avenue, 1001, Lavras, Minas Gerais State, 37200-000, Brazil

  2. Guilherme Oliveira

    Present address: Instituto Vale de Tecnologia, Rua Boaventura da Silva, 955, Belém, Pará, 66055-090, Brazil

Authors and Affiliations

  1. Department of Sanitary and Environmental Engineering, Federal University of Minas Gerais, Antonio Carlos Avenue, 6627, Belo Horizonte, Minas Gerais State, 31270-901, Brazil

    Luciene Alves Batista Siniscalchi, Carlos Augusto Lemos Chernicharo & Juliana Calabria de Araújo

  2. Department of Engineering, Federal University of Lavras, Dr. Sylvio Menicucci Avenue, 1001, Lavras, Minas Gerais State, 37200-000, Brazil

    Laura Rabelo Leite

  3. Genomics and Computational Biology Group, René Rachou Research Center, Oswaldo Cruz Foundation, Augusto de Lima Avenue 1715, Belo Horizonte, Minas Gerais State, 30.190-002, Brazil

    Guilherme Oliveira

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  1. Luciene Alves Batista Siniscalchi
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Correspondence to Juliana Calabria de Araújo.

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Siniscalchi, L.A.B., Leite, L.R., Oliveira, G. et al. Illumina sequencing-based analysis of a microbial community enriched under anaerobic methane oxidation condition coupled to denitrification revealed coexistence of aerobic and anaerobic methanotrophs. Environ Sci Pollut Res 24, 16751–16764 (2017). https://doi.org/10.1007/s11356-017-9197-9

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  • DOI: https://doi.org/10.1007/s11356-017-9197-9

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