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

Barret, Maialen and Gagnon, Nathalie and Kalmokoff, Martin L. and Topp, Edward and Verasteguy, Yris and Brooks, Stephen P. J. and Matias, Fernando and Neufeld, Josh D. and Talbot, Guylaine Identification of Methanoculleus spp. as active methanogens during anoxic incubations of swine manure storage tank samples. (2013) Applied and Environmental Microbiology, 79 (2). 424-433. ISSN 0099-2240

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Official URL: http://dx.doi.org/10.1128/AEM.02268-12

Abstract

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-¹³C] 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 ¹³C into DNA was detectable at in situ acetate concentrations (~7g/liter). Fingerprints of DNA retrieved from heavy fractions of the ¹³C 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.

Item Type:Article
Additional Information:Thanks to American Society for Microbiology editor. The original PDF can be found at http://aem.asm.org/content/79/2/424
HAL Id:hal-00917867
Audience (journal):International peer-reviewed journal
Uncontrolled Keywords:
Institution:Other partners > Agriculture and Agri-Food - AAFC (CANADA)
Other partners > Natural Sciences and Engineering Research Council of Canada - NSERC (CANADA)
Other partners > University of Waterloo (CANADA)
Funders:
Agriculture and Agri-Food Canada - National Sciences and Engineering Research Council of Canada
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Deposited On:12 Dec 2013 14:48

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