title: Temporal dynamics of fibrolytic and methanogenic rumen microorganisms during in situ incubation of switchgrass determined by 16S rRNA gene profiling.
abstract: The rumen microbial ecosystem is known for its biomass-degrading and methane-producing phenotype. Fermentation of recalcitrant plant material, comprised of a multitude of interwoven fibers, necessitates the synergistic activity of diverse microbial taxonomic groups that inhabit the anaerobic rumen ecosystem. Although interspecies hydrogen (H2) transfer, a process during which bacterially generated H2 is transferred to methanogenic Archaea, has obtained significant attention over the last decades, the temporal variation of the different taxa involved in in situ biomass-degradation, H2 transfer and the methanogenesis process remains to be established. Here we investigated the temporal succession of microbial taxa and its effect on fiber composition during rumen incubation using 16S rRNA amplicon sequencing. Switchgrass filled nylon bags were placed in the rumen of a cannulated cow and collected at nine time points for DNA extraction and 16S pyrotag profiling. The microbial community colonizing the air-dried and non-incubated (0 h) switchgrass was dominated by members of the Bacilli (recruiting 63% of the pyrotag reads). During in situ incubation of the switchgrass, two major shifts in the community composition were observed: Bacilli were replaced within 30 min by members belonging to the Bacteroidia and Clostridia, which recruited 34 and 25% of the 16S rRNA reads generated, respectively. A second significant shift was observed after 16 h of rumen incubation, when members of the Spirochaetes and Fibrobacteria classes became more abundant in the fiber-adherent community. During the first 30 min of rumen incubation ~13% of the switchgrass dry matter was degraded, whereas little biomass degradation appeared to have occurred between 30 min and 4 h after the switchgrass was placed in the rumen. Interestingly, methanogenic members of the Euryarchaeota (i.e., Methanobacteria) increased up to 3-fold during this period of reduced biomass-degradation, with peak abundance just before rates of dry matter degradation increased again. We hypothesize that during this period microbial-mediated fibrolysis was temporarily inhibited until H2 was metabolized into CH4 by methanogens. Collectively, our results demonstrate the importance of inter-species interactions for the biomass-degrading and methane-producing phenotype of the rumen microbiome-both microbially facilitated processes with global significance.
authors.affiliation: Systems Microbiology and Biotechnology Group, School of Molecular Biosciences, Washington State University Richland, WA, USA.
authors.forename: Hailan
authors.initials: H
authors.lastname: Piao
authors.affiliation: Department of Animal and Range Sciences, Montana State University Bozeman, MT, USA.
authors.forename: Medora
authors.initials: M
authors.lastname: Lachman
authors.affiliation: Lawrence Livermore National Laboratory, Biosciences and Biotechnology Division Livermore, CA, USA.
authors.forename: Stephanie
authors.initials: S
authors.lastname: Malfatti
authors.affiliation: Faculty of Technology and Center for Biotechnology, Bielefeld University Bielefeld, Germany.
authors.forename: Alexander
authors.initials: A
authors.lastname: Sczyrba
authors.affiliation: Lawrence Berkeley National Laboratory, Life Sciences Division Berkeley, CA, USA.
authors.forename: Bernhard
authors.initials: B
authors.lastname: Knierim
authors.affiliation: Lawrence Berkeley National Laboratory, Life Sciences Division Berkeley, CA, USA.
authors.forename: Manfred
authors.initials: M
authors.lastname: Auer
authors.affiliation: Prokaryote Super Program, DOE Joint Genome Institute Walnut Creek, CA, USA.
authors.forename: Susannah G
authors.initials: SG
authors.lastname: Tringe
authors.affiliation: Department of Animal Sciences and Institute for Genomic Biology, University of Illinois, Urbana-Champaign IL, USA.
authors.forename: Roderick I
authors.initials: RI
authors.lastname: Mackie
authors.affiliation: Department of Animal and Range Sciences, Montana State University Bozeman, MT, USA.
authors.forename: Carl J
authors.initials: CJ
authors.lastname: Yeoman
authors.affiliation: Systems Microbiology and Biotechnology Group, School of Molecular Biosciences, Washington State University Richland, WA, USA ; Prokaryote Super Program, DOE Joint Genome Institute Walnut Creek, CA, USA ; Energy and Efficiency Division, Chemical and Biological Process Development Group, Pacific Northwest National Laboratory Richland, WA, USA.
authors.forename: Matthias
authors.initials: M
authors.lastname: Hess
keywords: cellulolytic bacteria; interspecies H2 transfer; methanogenic archaea; microbe-microbe interactions; rumen microbiology
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