The microbial communities that produce biogas as a result of anaerobic digestion of manure remain poorly understood. Using next-generation sequencing, methanogen populations were investigated in three full scale mesophilic anaerobic digesters operated on dairy farms. A combined 50 246 non-chimeric sequence reads covering the V1-V3 hypervariable regions of the methanogen 16S rRNA gene were assigned to 307 species-level operational taxonomic units (OTUs). The Blue Spruce Farms (BSF) and Green Mountain Dairy (GMD) anaerobic digesters were found to have nearly identical methanogen profiles, with the overwhelming predominance of OTU 1 (98.5% and 99.7%, respectively), which showed 99.2% sequence identity to Methanosarcina thermophila. In contrast, methanogens from the Chaput Family Farms (CFF) anaerobic digester were more diverse, with five major OTUs belonging to four distinct phylogenetic groups (Methanomicrobiales, Methanosarcinales, Methanoplasmatales, and Methanobacteriales). Differences in management practices and years of operation were hypothesized as potential factors responsible for differences in the methanogen profiles. (C) 2013 Elsevier Ltd. All rights reserved.