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This 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 NH3-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 NH3-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. Funding Agencies|Swedish Energy Agency; Biogas Research Centre at Linkoping University, Sweden; NSF [DGE-1256082]

Aquatic fungi and bacteria have long been recognized as key drivers in ecosystem processes such as leaf litter decomposition. We examined fungal and bacterial communities on decaying alder Alnus gl ...

Addition of Co and Ni often improves the production of biogas during digestion of organic matter, i.e. increasing CH4-production, process stability and substrate utilization which often opens for higher organic loading rates (OLRs). The effect of Co and Ni addition was evaluated by measuring methane production, volatile solids reduction, pH and concentration of volatile fatty acids (VFAs). A series of six lab.-scale semi-continuously fed biogas tank reactors were used for this purpose. The chemical forms and potential bioavailability of Co and Ni were examined by sequential extraction, acid volatile sulfide extraction (AVS) and simultaneously extracted metals. Furthermore, the sulfur speciation in solid phase was examined by sulfur X-ray absorption near edge structure spectroscopy. The effect of Co and Ni deficiency on the microbial community composition was analyzed using quantitative polymerase chain reaction and 454-pyrosequencing. The results showed that amendment with Co and Ni was necessary to maintain biogas process stability and resulted in increased CH4-production and substrate utilization efficiency. 10–20% of the total Co concentration was in dissolved form and should be regarded as easily accessible by the microorganisms. In contrast, Ni was entirely associated with organic matter/sulfides (mainly AVS) and regarded as very difficult to take up. Still Ni had stimulatory effects suggesting mechanisms such as dissolution of NiS to be involved in the regulation of Ni availability for the microorganisms. The microbial community structure varied in relation to the occurrence of Ni and Co. The acetate-utilizing Methanosarcinales dominated during stable process performance, i.e. when both Co and Ni were supplied, while hydrogenotrophic Methanomicrobiales increased together with VFA concentrations under Co or Ni deficiency. The increase was more pronounced at Co limitation. This study demonstrates that there are good possibilities to improve the performance of biogas processes digesting sulfur-rich substrates by supplementation of Co and Ni.

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 160000 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.

The effect of trace element addition on anaerobic digestion of food industry- and household waste was studied using two semi-continuous lab-scale reactors, one (R30+) was supplied with Fe, Co and Ni, while the other (R30) acted as a control. Tracer analysis illustrated that methane production from acetate proceeded through syntrophic acetate oxidation (SAO) in both digesters. The effect of the trace elements was also evaluated in batch assays to determine the capacity of the microorganisms of the two digesters to degrade acetate, phenyl acetate, oleic acid or propionate, butyrate and valerate provided as a cocktail. The trace elements addition improved the performance of the process giving higher methane yields during start-up and early operation and lower levels of mainly acetate and propionate in the R30+ reactor. The batch assay showed that material from R30+ gave effects on methane production from all substrates tested. Phenyl acetate was observed to inhibit methane formation in the R30 but not in the R30+ assay. A real-time PCR analysis targeting methanogens on the order level as well as three SAO bacteria showed an increase in Methanosarcinales in the R30+ reactor over time, even though SAO continuously was the dominating pathway for methane production. Possibly, this increase explains the low VFA-levels and higher degradation rates observed in the R30+ batch incubations. These results show that the added trace elements affected the ability of the microflora to degrade VFAs as well as oleic acid and phenyl acetate in a community, where acetate utilization is dominated by SAO.

This review summarizes the microbial mechanisms responsible for removal of carbon, nitrogen. and sulfur compounds in treatment wetlands (TWs) and identifies, categorizes and compares various techni ...

Constructed wetlands can be used to decrease the high ammonium concentrations in landfill leachates. We investigated nitrification/denitrification activity and the corresponding bacterial communities in landfill leachate that was treated in a compact constructed wetland, Tveta Recycling Facility, Sweden. Samples were collected at three depths in a filter bed and the sediment from a connected open pond in July, September and November 2004. Potential ammonia oxidation was measured by short-term incubation method and potential denitrification by the acetylene inhibition technique. The ammonia-oxidising and the denitrifying bacterial communities were investigated using group-specific PCR primers targeting 16S rRNA genes and the functional gene nosZ, respectively. PCR products were analysed by denaturing gradient gel electrophoresis and nucleotide sequencing. The same degree of nitrification activity was observed in the pond sediment and at all levels in the filter bed, whereas the denitrification activity decreased with filter bed depth. Denitrification rates were higher in the open pond, even though the denitrifying bacterial community was more diverse in the filter bed. The ammonia-oxidising community was also more varied in the filter bed. In the filter bed and the open pond, there was no obvious relationship between the nitrification/denitrification activities and the composition of the corresponding bacterial communities.

Overland flow systems are useful for treating landfill leachates, because they provide favourable conditions for nitrification and they are easy to maintain. However, little is known about the microbial communities in such systems or the nitrification capacity of those microorganisms. In this study, seasonal variations in potential nitrification and in community composition of nitrifying bacteria were investigated in two overland flow areas receiving leachate from landfills at Korslot and Hagby, Sweden. Samples were collected in the settling ponds sediment and at two depths in the overland flow areas (the macrophyte litter layer and the rhizosphere) in May, August and November 2003. A short-term incubation method was used to measure potential oxidation of ammonia and nitrite (designated PAO and PNO). The ammonia-oxidising bacterial (AOB) community was investigated using a 16S rRNA gene approach that included PCR amplification and analysis of PCR products by denaturing gradient gel electrophoresis (DGGE), followed by nucleotide sequencing and phylogenetic analysis. PAO was determined in the range 5–2700 (NO 2 − +NO 3 − )-N g −1 dw d −1 and PNO in the range 60–2000 μg NO 2 − -N g −1 dw d −1 . At Korslot, PAO and PNO showed similar temporal variation in the different ecosystems, whereas no such relationship was noticed at Hagby. Considering both sites, there was no obvious change in the composition of the AOB community over the growing season. However, the composition did differ between the ecosystems: Nitrosomonas -like sequences were more common in the ponds, and in the litter layers they were found as often as Nitrosospira -like sequences, whereas Nitrosospira -like sequences were more common in the rhizospheres. Altogether, we found nine different AOB sequences, five Nitrosomonas -like and four Nitrosospira -like, which belonged to clusters 0, 2, 3b, 6a, 6b and 7. There was no apparent relationship between the number of AOB populations and the PAO in different soil layers and sediments.

This study used semi-continuous laboratory scale biogas reactors to simulate the effects of trace-element addition in different combinations, while degrading the organic fraction of municipal solid waste and slaughterhouse waste. The results show that the combined addition of Fe, Co and Ni was superior to the addition of only Fe, Fe and Co or Fe and Ni. However, the addition of only Fe resulted in a more stable process than the combined addition of Fe and Co, perhaps indicating a too efficient acidogenesis and/or homoacetogenesis in relation to a Ni-deprived methanogenic population. The results were observed in terms of higher biogas production (+9%), biogas production rates (+35%) and reduced VFA concentration for combined addition compared to only Fe and Ni. The higher stability was supported by observations of differences in viscosity, intraday VFA- and biogas kinetics as well as by the 16S rRNA gene and 16S rRNA of the methanogens.