Identification of Methanoculleus spp. as active methanogens during anoxic incubations of swine manure storage tank samples

Autor: Fernando Matias, Josh D. Neufeld, Nathalie Gagnon, Yris Verastegui, Maialen Barret, Martin Kalmokoff, Stephen P. J. Brooks, Edward Topp, Guylaine Talbot
Přispěvatelé: Agriculture and Agri-Food - AAFC (CANADA), Natural Sciences and Engineering Research Council of Canada - NSERC (CANADA), University of Waterloo (CANADA), Agriculture and Agri-Food [Ottawa] (AAFC), University of Waterloo [Waterloo]
Jazyk: angličtina
Rok vydání: 2013
Předmět:
[SDV.SA]Life Sciences [q-bio]/Agricultural sciences
Manure management
Swine
Methanogenesis
Microorganism
Stable isotope probing
Molecular Sequence Data
Stable-isotope probing
DNA
Ribosomal
Polymerase Chain Reaction
Applied Microbiology and Biotechnology
RNA
Ribosomal
16S
Environmental Microbiology
Animals
Cluster Analysis
Anaerobiosis
Food science
Phylogeny
Ecology
biology
Sequence Analysis
DNA
Science des productions animales
biology.organism_classification
Manure
Anoxic waters
Archaea
Microbiologie et Parasitologie
DNA
Archaeal
Methanoculleus
[SDV.MP]Life Sciences [q-bio]/Microbiology and Parasitology
Biochemistry
Isotope Labeling
[SDV.SA.SPA]Life Sciences [q-bio]/Agricultural sciences/Animal production studies
Methanomicrobiaceae
Oxidoreductases
Methane
Sciences agricoles
Food Science
Biotechnology
Zdroj: Applied and Environmental Microbiology
Applied and Environmental Microbiology, American Society for Microbiology, 2013, vol. 79, pp. 424-433. ⟨10.1128/AEM.02268-12⟩
ISSN: 0099-2240
1098-5336
DOI: 10.1128/AEM.02268-12⟩
Popis: Methane emissions represent a major environmental concern associated with manure management in the livestock industry. A more thorough understanding of how microbial communities function in manure storage tanks is a prerequisite for mitigating methane emissions. Identifying the microorganisms that are metabolically active is an important first step. Methanogenic archaea are major contributors to methanogenesis in stored swine manure, and we investigated active methanogenic populations by DNA stable isotope probing (DNA-SIP). Following a preincubation of manure samples under anoxic conditions to induce substrate starvation, [U- 13 C]acetate was added as a labeled substrate. Fingerprint analysis of density-fractionated DNA, using length-heterogeneity analysis of PCR-amplified mcrA genes (encoding the alpha subunit of methyl coenzyme M reductase), showed that the incorporation of 13 C into DNA was detectable at in situ acetate concentrations (∼7 g/liter). Fingerprints of DNA retrieved from heavy fractions of the 13 C treatment were primarily enriched in a 483-bp amplicon and, to a lesser extent, in a 481-bp amplicon. Analyses based on clone libraries of the mcrA and 16S rRNA genes revealed that both of these heavy DNA amplicons corresponded to Methanoculleus spp. Our results demonstrate that uncultivated methanogenic archaea related to Methanoculleus spp. were major contributors to acetate-C assimilation during the anoxic incubation of swine manure storage tank samples. Carbon assimilation and dissimilation rate estimations suggested that Methanoculleus spp. were also major contributors to methane emissions and that the hydrogenotrophic pathway predominated during methanogenesis.
Databáze: OpenAIRE
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