title: Influences of the substrate feeding regime on methanogenic activity in biogas reactors approached by molecular and stable isotope methods.
abstract: In order to better understand the effects of the substrate feeding regime on methanogenesis during anaerobic digestion in biogas reactors, four continuous stirred tank reactors operated under mesophilic conditions were investigated. In addition to standard physicochemical parameters, the stable isotopic signatures of CH4 and CO2 before and after daily feeding were analyzed. The activity of the methanogens was assessed by methyl coenzyme M reductase alpha-subunit (mcrA/mrtA) gene transcript analysis. Two different feeding regimes i.e. single vs. double consecutive feeding of the otherwise same daily maize silage load were investigated. During the first phase, a single feeding of the whole daily dose increased the biogas production within 70-80 min from around 0.5 to 2.0 L/h. This increase was associated with a transient increase of the acetic acid concentration and a corresponding decrease of the pH. Only moderate increase in biogas yield and VFA concentration (mainly acetate) was observed when the daily substrate was apportioned into two feedings. However, the overall daily gas production was similar in both cases. Regardless of the feeding regime, significantly depleted δ(13)CH4 and minor changes in the CO2 content of biogas were observed after feeding, which were followed by enrichment of δ(13)CH4. This period was associated with detectable changes in activity of methanogenic communities monitored by terminal restriction fragment length polymorphism analysis based on the transcripts of mcrA/mrtA genes. Methanoculleus and Methanobacterium spp. were the predominant methanogens in all reactors, while Methanosarcina spp. activity was only significant in two reactors. The activity of Methanoculleus and Methanosarcina spp. increased after the feeding in these reactors, which was followed by a depletion of δ(13)C in the produced gas. In both reactors, the less depleted isotopic values were detected before the second feeding, when Methanobacterium was the most active genus. Variations in reactor performance and methanogenic community characteristics were attributed to inoculum heterogeneity and stochastic factors during the reactor set up.
authors.affiliation: Department of Bioenergy, Helmholtz Centre for Environmental Research - UFZ, Permoserstrasse 15, 04318 Leipzig, Germany.
authors.forename: Z
authors.initials: Z
authors.lastname: Lv
authors.affiliation: Department of Bioenergy, Helmholtz Centre for Environmental Research - UFZ, Permoserstrasse 15, 04318 Leipzig, Germany.
authors.forename: A F
authors.initials: AF
authors.lastname: Leite
authors.affiliation: Department of Environmental Microbiology, Helmholtz Centre for Environmental Research - UFZ, Permoserstrasse 15, 04318 Leipzig, Germany.
authors.forename: H
authors.initials: H
authors.lastname: Harms
authors.affiliation: Department of Isotope Biogeochemistry, Helmholtz Centre for Environmental Research - UFZ, Permoserstrasse 15, 04318 Leipzig, Germany.
authors.forename: H H
authors.initials: HH
authors.lastname: Richnow
authors.affiliation: Department of Biochemical Conversion, Deutsches Biomasseforschungszentrum (DBFZ), Torgauer Str. 116, 04347 Leipzig, Germany.
authors.forename: J
authors.initials: J
authors.lastname: Liebetrau
authors.affiliation: Department of Bioenergy, Helmholtz Centre for Environmental Research - UFZ, Permoserstrasse 15, 04318 Leipzig, Germany. Electronic address: marcell.nikolausz@ufz.de.
authors.forename: M
authors.initials: M
authors.lastname: Nikolausz
mesh_terms: D061249:Batch Cell Culture Techniques / Q000379:methods; D056804:Biofuels; D015415:Biomarkers / Q000378:metabolism; D019149:Bioreactors; D002245:Carbon Dioxide / Q000378:metabolism; D002247:Carbon Isotopes; D019605:Euryarchaeota / Q000145:classification / Q000235:genetics* / Q000378:metabolism; D006863:Hydrogen-Ion Concentration; D008697:Methane / Q000096:biosynthesis*; D059013:Microbial Consortia / Q000235:genetics*; D010088:Oxidoreductases / Q000235:genetics*; D011312:Pressure; D060888:Real-Time Polymerase Chain Reaction; D013696:Temperature; D003313:Zea mays / Q000378:metabolism*
chemical_list: D056804:Biofuels; D015415:Biomarkers; D002247:Carbon Isotopes; D002245:Carbon Dioxide; D010088:Oxidoreductases; C021572:methyl coenzyme M reductase; D008697:Methane
keywords: Biogas; Methanogenic activity; Molecular tools; Stable isotope approach; mcrA; mrtA