Anaerobic degradation of protein-rich materials has high methane potential and produces nutrient-rich residue, but requires strategies to avoid ammonia inhibition. A well-adapted process can cope with substantially higher ammonia levels than an unadapted process and analyses of pathways for methanisation of acetate, combined with determination of microbial community structure, strongly indicate that this is due to a significant contribution of syntrophic acetate oxidation. The microorganisms involved in syntrophic acetate oxidation thus most likely occupy a unique niche and play an important role in methane formation. This review summarises current insight of syntrophic acetate oxidising microorganisms, their presence and the detection of novel species and relate these observations with operating conditions of the biogas processes in order to explore contributing factors for development of an ammonia-tolerant microbial community that efficiently degrades acetate through the syntrophic pathway. Besides high ammonia level, acetate concentration, temperature and methanogenic community structure are considered in this review as likely factors that shape and influence SAO-mediated microbial ecosystems. The main purpose of this review is to facilitate process optimisation through considering the activity and growth of this key microbial community

Methane (CH4) is produced as an end substrate from feed degradation in the rumen. Yield of CH4 varies between individuals despite identical feeding conditions. To get a better understanding of factors behind the individual variation, 73 dairy cows given the same feed but differing in CH4 emissions were investigated with focus on fiber digestion, fermentation end products and bacterial and archaeal composition. In total 21 cows (12 Holstein, 9 Swedish Red) identified as persistent low, medium or high CH4 emitters over a three month period were furthermore chosen for analysis of microbial community structure in rumen fluid. This was assessed by sequencing the V4 region of 16S rRNA gene and by quantitative qPCR of targeted Methanobrevibacter groups. The results showed a positive correlation between low CH4 emitters and higher abundance of Methanobrevibacter ruminantium clade. Principal coordinate analysis (PCoA) on operational taxonomic unit (OTU) level of bacteria showed two distinct clusters (P<0.01) that was related to CH4 production. One cluster was associated with low CH4 production (referred to as cluster L) whereas the other cluster was associated with high CH4 production (cluster H) and the medium emitters occurred in both clusters. The differences between clusters were primarily linked to differential abundances of certain OTUs belonging to Prevotella. Moreover, several OTUs belonging to the family Succinivibrionaceae were dominant in samples belonging to cluster L. Fermentation pattern of volatile fatty acids showed that proportion of propionate was higher in cluster L, while proportion of butyrate was higher in cluster H. No difference was found in milk production or organic matter digestibility between cows. Anyhow, cows in cluster L had lower CH4/ kg energy corrected milk (ECM) compared to cows in cluster H, 8.3 compared to 9.7 g CH4/kg ECM, showing that low CH4 cows utilized the feed more efficient for milk production which might indicate a more efficient microbial population or host genetic differences that is reflected in microbial population.

In biogas processes, methane production from acetate proceeds by either aceticlastic methanogenesis or through syntrophic acetate oxidation (SAO). In the present study, the pathway for methane production from acetate was analysed; i) during a gradual increase of the ammonia concentration (final concentration 7 g NH4+−N/L) in a semi-continuous lab-scale anaerobic digester (4.3 L), operating at mesophilic temperature (37°C) or ii) in diluted enrichment cultures (100 ml) experiencing a gradual increase in ammonia, sodium, potassium and propionic acid. The pathway for methane formation was determined by calculating the 14CO2/14CH4 ratio after incubating samples with 14C-2-acetate. In the anaerobic digester, as well as in the enrichment cultures, the 14CO2/14CH4 ratio clearly increased with increasing ammonium-nitrogen concentration, i.e. as the ammonia concentration increased, a shift from the aceticlastic mechanism to the syntrophic pathway occurred. The shift was very distinct and occurred as the NH4+−N concentration rose above 3 g/l. No shift in pathway was seen during increasing concentrations of sodium, potassium or propionic acid. The shift to SAO in the biogas digester resulted in a twofold decrease in the specific gas and methane yield.

The bacterial and archaeal community structure was examined in two methanogenic anaerobic digestion processes degrading organic household waste at me

A syntrophic acetate-oxidizing bacterium, strain BST (T = type strain), was isolated from a previously described mesophilic triculture that was able to syntrophically oxidize acetate and form methane in stoichiometric amounts. Strain BST was isolated with substrates typically utilized by homoacetogenic bacteria. Strain BST was a spore-forming, gram-positive, rod-shaped organism which utilized formate, glucose, ethylene glycol, cysteine, betaine, and pyruvate. Acetate and sometimes formate were the main fermentation products. Small amounts of alanine were also produced from glucose, betaine, and cysteine. Strain BST grew optimally at 37 degrees C and pH 7. The G+C content of the DNA of strain BST was 32 mol%. A 16S rRNA sequence analysis revealed that strain BST was a member of a new species of the genus Clostridium. We propose the name Clostridium ultunense for this organism; strain BS is the type strain of C. ultunense.

The growing number of biogas plants in Europe has resulted in increased production of nutrient-rich digestate with great potential as fertilizer for arable land. The nutrient composition of digestate varies with the substrate treated in the biogas plant and may contain compounds that stimulate or inhibit soil microbial activity. This study compared 20 digestates (D) with 10 pig slurries (PS) and 10 cow manures (CM) regarding their chemical content and their effect on soil microbial activities, i.e. potential ammonia oxidation rate (PAO) and soil respiration. The results showed no significant differences within the D group when divided based on substrate type. i.e. manure dominated vs. other organic waste materials in any of the tests. In general D contained significantly higher concentrations of ammonium while the concentrations of total carbon and volatile fatty acids were higher in PS and CM than in D. The D showed both stimulating and inhibiting effects on PAO, while all CM and all PS except one showed inhibiting effects on PAO. However, PAO activity was negatively correlated with the content of volatile fatty acids in the residues indicating that these compounds may be the cause of the inhibition. The maximum respiration activity (hpeakmax) was lower and the time point for the maximum respiration activity (tpeakmax) occurred earlier for D compared with CM and PS. This earlier peak time could be indicative of a high proportion of easily degradable carbon in D compared with PS and CM. However, the utilization rate of carbon, i.e. the proportion of added organic C converted to CO2-C during 12days, did not differ significantly between D, PS and CM, indicating that overall carbon quality in the different fertilizers was still roughly comparable. In short, our results suggest that digestates were different compared with PS and CM but without posing a higher risk with respect to their impact on soil microbial activity.