title: Biphasic Study to Characterize Agricultural Biogas Plants by High-Throughput 16S rRNA Gene Amplicon Sequencing and Microscopic Analysis.
abstract: Process surveillance within agricultural biogas plants (BGPs) was concurrently studied by high-throughput 16S rRNA gene amplicon sequencing and an optimized quantitative microscopic fingerprinting (QMF) technique. In contrast to 16S rRNA gene amplicons, digitalized microscopy is a rapid and cost-effective method that facilitates enumeration and morphological differentiation of the most significant groups of methanogens regarding their shape and characteristic autofluorescent factor 420. Moreover, the fluorescence signal mirrors cell vitality. In this study, four different BGPs were investigated. The results indicated stable process performance in the mesophilic BGPs and in the thermophilic reactor. Bacterial subcommunity characterization revealed significant differences between the four BGPs. Most remarkably, the generaDefluviitogaandHalocelladominated the thermophilic bacterial subcommunity, whereas members of another taxon,Syntrophaceticus, were found to be abundant in the mesophilic BGP. The domain Archaea was dominated by the genusMethanoculleusin all four BGPs, followed byMethanosaetain BGP1 and BGP3. In contrast,Methanothermobactermembers were highly abundant in the thermophilic BGP4. Furthermore, a high consistency between the sequencing approach and the QMF method was shown, especially for the thermophilic BGP. The differences elucidated that using this biphasic approach for mesophilic BGPs provided novel insights regarding disaggregated single cells ofMethanosarcinaandMethanosaetaspecies. Both dominated the archaeal subcommunity and replaced coccoidMethanoculleusmembers belonging to the same group ofMethanomicrobialesthat have been frequently observed in similar BGPs. This work demonstrates that combining QMF and 16S rRNA gene amplicon sequencing is a complementary strategy to describe archaeal community structures within biogas processes.
authors: Bielefeld University, Center for Biotechnology (CeBiTec), Genome Research and Systems Biology, 33615 Bielefeld, Germany.
authors: Irena
authors: I
authors: Maus
authors: Hamburg University of Applied Sciences (HAW), Faculty Life Sciences / Research Center "Biomass Utilization Hamburg", Laboratory for Applied Microbiology, 21033 Hamburg-Bergedorf, Germany.
authors: Yong Sung
authors: YS
authors: Kim
authors: Bielefeld University, Center for Biotechnology (CeBiTec), Genome Research and Systems Biology, 33615 Bielefeld, Germany.
authors: Daniel
authors: D
authors: Wibberg
authors: Bielefeld University, Center for Biotechnology (CeBiTec), Genome Research and Systems Biology, 33615 Bielefeld, Germany.
authors: Yvonne
authors: Y
authors: Stolze
authors: Hamburg University of Applied Sciences (HAW), Faculty Life Sciences / Research Center "Biomass Utilization Hamburg", Laboratory for Applied Microbiology, 21033 Hamburg-Bergedorf, Germany.
authors: Sandra
authors: S
authors: Off
authors: Hamburg University of Applied Sciences (HAW), Faculty Life Sciences / Research Center "Biomass Utilization Hamburg", Laboratory for Applied Microbiology, 21033 Hamburg-Bergedorf, Germany.
authors: Sebastian
authors: S
authors: Antonczyk
authors: Bielefeld University, Center for Biotechnology (CeBiTec), Genome Research and Systems Biology, 33615 Bielefeld, Germany.
authors: Alfred
authors: A
authors: Pühler
authors: Hamburg University of Applied Sciences (HAW), Faculty Life Sciences / Research Center "Biomass Utilization Hamburg", Laboratory for Applied Microbiology, 21033 Hamburg-Bergedorf, Germany.
authors: Paul
authors: P
authors: Scherer
authors: Bielefeld University, Center for Biotechnology (CeBiTec), Genome Research and Systems Biology, 33615 Bielefeld, Germany.
authors: Andreas
authors: A
authors: Schlüter
mesh_terms: D000383:Agriculture; D000693:Anaerobiosis; D001105:Archaea / Q000145:classification* / Q000235:genetics / Q000302:isolation & purification* / Q000648:ultrastructure; D056804:Biofuels / Q000382:microbiology; D019149:Bioreactors; D016172:DNA Fingerprinting / Q000379:methods; D020459:Genes, rRNA; D059014:High-Throughput Nucleotide Sequencing; D017013:Methanobacteriaceae / Q000235:genetics / Q000302:isolation & purification; D059013:Microbial Consortia / Q000235:genetics; D008853:Microscopy; D010802:Phylogeny; D012336:RNA, Ribosomal, 16S / Q000235:genetics; D012722:Sewage / Q000032:analysis / Q000382:microbiology*
chemical_list: D056804:Biofuels; D012336:RNA, Ribosomal, 16S; D012722:Sewage
keywords: Anaerobic digestion; archaeal morphology; microscopy; taxonomic profile