The microbial community of 21 full-scale biogas reactors was examined using 454 pyrosequencing of 16S rRNA gene sequences. These reactors included seven (six mesophilic and one thermophilic) digesting sewage sludge (SS) and 14 (ten mesophilic and four thermophilic) codigesting (CD) various combinations of wastes from slaughterhouses, restaurants, households, etc. The pyrosequencing generated more than 160,000 sequences representing 11 phyla, 23 classes, and 95 genera of Bacteria and Archaea. The bacterial community was always both more abundant and more diverse than the archaeal community. At the phylum level, the foremost populations in the SS reactors included Actinobacteria, Proteobacteria, Chloroflexi, Spirochetes, and Euryarchaeota, while Firmicutes was the most prevalent in the CD reactors. The main bacterial class in all reactors was Clostridia. Acetoclastic methanogens were detected in the SS, but not in the CD reactors. Their absence suggests that methane formation from acetate takes place mainly via syntrophic acetate oxidation in the CD reactors. A principal component analysis of the communities at genus level revealed three clusters: SS reactors, mesophilic CD reactors (including one thermophilic CD and one SS), and thermophilic CD reactors. Thus, the microbial composition was mainly governed by the substrate differences and the process temperature.

With the final aim of reducing the energy consumption and increase the methane production at Swedish pulp and paper mills, the methane potential of 62 wastewater effluents from ten processes at seven pulp and/or paper mills (A-G) was determined in anaerobic batch digestion assays. This mapping is a first step towards an energy efficient and more sustainable utilization of the effluents by anaerobic digestion, and will be followed up by tests in lab-scale and pilot-scale reactors. Five of the mills produce kraft pulp (KP), one thermo-mechanical pulp (TMP), two chemical thermo-mechanical pulp (CTMP) and two neutral sulfite semichemical (NSSC) pulp. Both elemental and total chlorine free (ECF and TCF, respectively) bleaching processes were included. The effluents included material from wood rooms, cooking and oxygen delignification, bleaching (often both acid-and alkali effluents), drying and paper/board machinery as well as total effluents before and after sedimentation.The results from the screening showed a large variation in methane yields (percent of theoretical methane potential assuming 940 NmL CH 4 per g TOC) among the effluents. For the KP-mills, methane yields above 50% were obtained for the cooking effluents from mills D and F, paper machine wastewater from mill D, condensate streams from mills B, E and F and the composite pre-sedimentation effluent from mill D. The acidic ECF-effluents were shown to be the most toxic to the AD-flora and also seemed to have a negative effect on the yields of composite effluents downstream while three of the alkaline ECF-bleaching effluents gave positive methane yields. ECF bleaching streams gave higher methane yields when hardwood was processed. All TCF-bleaching effluents at the KP mills gave similar degradation patterns with final yields of 10-15% of the theoretical methane potential for four of the five effluents. The composite effluents from the two NSSC-processes gave methane yields of 60% of the theoretical potential.The TMP mill (A) gave the best average yield with all 6 effluents ranging 40-65% of the theoretical potential. The three samples from the CTMP process at mill B showed potentials around 40% while three of the six effluents at mill G (CTMP) yielded 45-50%.

SummaryThis study examined whether the abundance and expression of microbial 16S rRNA genes were associated with elemental concentrations and substrate conversion biokinetics in 20 full‐scale anaerobic digesters, including seven municipal sewage sludge (SS) digesters and 13 industrial codigesters. SS digester contents had higher methane production rates from acetate, propionate and phenyl acetate compared to industrial codigesters. SS digesters and industrial codigesters were distinctly clustered based on their elemental concentrations, with higher concentrations of NH 3‐N, Cl, K and Na observed in codigesters. Amplicon sequencing of 16S rRNA genes and reverse‐transcribed 16S rRNA revealed divergent grouping of microbial communities between mesophilic SS digesters, mesophilic codigesters and thermophilic digesters. Higher intradigester distances between Archaea 16S rRNA and rRNA gene profiles were observed in mesophilic codigesters, which also had the lowest acetate utilization biokinetics. Constrained ordination showed that microbial rRNA and rRNA gene profiles were significantly associated with maximum methane production rates from acetate, propionate, oleate and phenyl acetate, as well as concentrations of NH 3‐N, Fe, S, Mo and Ni. A co‐occurrence network of rRNA gene expression confirmed the three main clusters of anaerobic digester communities based on active populations. Syntrophic and methanogenic taxa were highly represented within the subnetworks, indicating that obligate energy‐sharing partnerships play critical roles in stabilizing the digester microbiome. Overall, these results provide new evidence showing that different feed substrates associate with different micronutrient compositions in anaerobic digesters, which in turn may influence microbial abundance, activity and function.

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Legionellales is an order of the Gammaproteobacteria, only composed of host-adapted, intracellular bacteria, including the accidental human pathogens Legionella pneumophila and Coxiella burnetii. Although the diversity in terms of lifestyle is large across the order, only a few genera have been sequenced, owing to the difficulty to grow intracellular bacteria in pure culture. In particular, we know little about their global distribution and abundance.Here, we analyze 16/18S rDNA amplicons both from tens of thousands of published studies and from two separate sampling campaigns in and around ponds and in a silver mine. We demonstrate that the diversity of the order is much larger than previously thought, with over 450 uncultured genera. We show that Legionellales are found in about half of the samples from freshwater, soil and marine environments and quasi-ubiquitous in man-made environments. Their abundance is low, typically 0.1%, with few samples up to 1%. Most Legionellales OTUs are globally distributed, while many do not belong to a previously identified species.This study sheds a new light on the ubiquity and diversity of one major group of host-adapted bacteria. It also emphasizes the need to use metagenomics to better understand the role of host-adapted bacteria in all environments.