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A novel facultatively anaerobic moderately thermophilic bacteria, strains 4137-MeT and 4148-MeT, were isolated from hot springs of Karmadon and Ursdon, respectively (North Ossetia, Russian Federation). Gram-negative, motile rods were present singly, in pairs, rosettes, and aggregates, or formed biofilms. Both strains grew optimally at 50-55 °C, pH 7.0 and did not require sodium chloride or yeast extract for growth. They were chemoorganoheterotrophs, growing on mono-, di- and polysaccharides (cellulose, starch, xylan, lichenan, galactan, xyloglucan, mannan, xanthan gum, guar gum) as well as proteinaceous substrates (gelatin, peptone, beef and yeast extract). Growth under anaerobic conditions was observed in presence and absence of external electron acceptors. Sulfur, thiosulfate, arsenate, Fe-citrate, and ferrihydrite were reduced with acetate, starch, or yeast extract as electron donors. The respiratory quinone was MK-7. Major cellular fatty acids of both strains were iso-C15:0, anteiso-C17:0, C15:0, iso-C16:0 and additionally iso-C17:0 for strain 4137-MeT. The size of the genome and genomic DNA G + C content of strain 4137-MeT were 3.24 Mb. and 29.9 %, respectively; for strain 4148-MeT - 3.33 Mb and 30.7 %. According to the 16S rRNA gene sequence and conserved protein sequences phylogenies, strains 4137-MeT and 4148-MeT represented a distinct lineage of the family Melioribacteraceae within the class Ignavibacteria. Based on phylogenetic analysis and phenotypic features, the novel isolates were assigned to a novel genus, for which the name Rosettibacter gen. nov. is proposed. Strain 4148-MeT represents its type species Rosettibacter primus sp. nov., while strain 4137-MeT represents a new species Rosettibacter firmus sp. nov.

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In anaerobic digestion (AD), the choice of inoculum type seems to be relevant for methane production for complex substrates, such as lignocellulosic material. Previous work demonstrated that the addition of fresh manure and ruminal fluid to anaerobic sludge improved methane productivity and kinetics of AD of crude sugarcane bagasse (CSB). Considering that the improvement of methane production could be a result of a more adapted microbial community, the present study performed the Next Generation Sequencing analysis to identify changes in the microbiome of anaerobic sludge inoculum, resulting from fresh manure and ruminal fluid addition. In comparison with AD performed only with anaerobic sludge inoculum (50:50, U), accumulated methane production was 15% higher with anaerobic sludge plus ruminal fluid inoculum (50:50, UR) and even higher (68%) with anaerobic sludge with fresh bovine manure inoculum (50:50, UFM), reaching the value of 143 NmLCH4.gVS-1. Clostridium species were highly abundant in all inocula, playing an important role during the hydrolysis and fermentation of CSB, and detoxifying potential inhibitors. Microbial composition also revealed the occurrence of Pseudomonas and Anaerobaculum at UFM inoculum that seem to have contributed to the higher methane production rate, mainly due to their hydrolytic and fermentative ability on lignocellulosic substrates. On the other hand, the presence of Alcaligenes might have had a negative effect on methane production due to their ability to perform methane oxidation.

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Bioaugmentation technology for improving the performance of thermophilic anaerobic digestion (TAD) of food waste (FW) treatment is gaining more attention. In this study, four thermophilic strains (Ureibacillus suwonensis E11, Clostridium thermopalmarium HK1, Bacillus thermoamylovorans Y25 and Caldibacillus thermoamylovorans QK5) were inoculated in the TAD of FW system, and the biochemical methane potential (BMP) batch study was conducted to assess the potential of different bioaugmented strains to enhance methane production. The results showed that the cumulative methane production in groups inoculated with E11, HK1, Y25 and QK5 improved by 2.05%, 14.54%, 19.79% and 9.17%, respectively, compared with the control group with no inoculation. Moreover, microbial community composition analysis indicated that the relative abundance of the main hydrolytic bacteria and/or methanogenic archaea was increased after bioaugmentation, and the four strains successfully became representative bacterial biomarkers in each group. The four strains enhanced methane production by strengthening starch, sucrose, galactose, pyruvate and methane metabolism functions. Further, the correlation networks demonstrated that the representative bacterial genera had positive correlations with the differential metabolic functions in each bioaugmentation group. This study provides new insights into the TAD of FW with bioaugmented strains.

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Hydrolytic bacteria are essential for the degradation of lignocellulose to produce biogas and organic fertilizers. In this study, sheep manure was used as substrate, and sheep manure slurry, yak rumen fluid and slurry from a biogas reactor (SBR) were used as inocula in single-stage anaerobic digestion. The SBR and rumen fluid inocula increased biogas production by 23% and 43%, respectively, when compared to solely sheep manure in the single-stage anaerobic digestion. The two-stage anaerobic digestion, with yak rumen fluid as inoculum in the hydrolytic reactor, increased the biogas production by 59, 86, and 58% compared with the control. Microbial analysis of the effluent revealed that yak rumen fluid contained hydrolytic bacteria such as Proteiniphilum, Jeotgalibaca, Fermentimonas, and Atopostipes to enhance the degradation of sheep manure and increase biogas production. It was concluded that yak rumen fluid, rich in hydrolytic bacteria, increases the degradability of sheep manure and improves production of volatile fatt acids and biogas.

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Anaerobic digestion (AD) is a widely applied technology for treating organic wastes to generate renewable energy in the form of biogas. The effectiveness of AD process depends on many factors, among which the most important is the presence of active and healthy microbial community in the anaerobic digesters, which needs to be explored. However, the deciphering of microbial populations and their functions during the AD process of different materials is still incomplete, which restricts the understanding of its long-term performance under different operational conditions. This review describes the type, morphology, functions, and specific growth conditions of commonly found hydrolytic, acidogenic, acetogenic bacteria, and archaea during the AD process. The effects of microbes on the performance and stability of the digestion process are also presented. Furthermore, the article offers a deep understanding of the AD management strategies for the enhancement of methane production and the efficiency of the energy conversion process of various organic wastes.

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Sludge, of which alginate-like biomaterial is a major organic component, is an increasing environmental problem. Thus, efficient anaerobic degradation of alginate provides a new method for sludge utilization. In this study, anaerobic alginate hydrolytic bacteria (AHB) were proposed to enrich with methanogens synergetically to reduce the inhibition of intermediate metabolites. The COD of produced methane reached 80.7 ±â€¯1.9% (n = 4) of initial alginate COD. After considering the microbial growth (8%-18% of COD), a good COD balance indicated that alginate was fully consumed and the main final metabolites were methane and CO2. Methanogenesis could promote alginate conversion by AHB. The enriched bacteria for alginate degradation in this study were different from that of former known AHB. The metabolic pathway of alginate degradation was revealed by metagenomics, in which oligo-alginate lyase was detected in twelve bacteria, and typical carbon metabolic pathways to convert alginate to methane were identified. More studies of bacterial isolation and biofuel production are still needed in the future.

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Cattle manure is frequently used as an inoculum for the start-up of agricultural biogas plants or as a co-substrate in the anaerobic digestion of lignocellulosic feedstock. Ruminal microbiota are considered to be effective plant fiber degraders, but the microbes contained in manure do not necessarily reflect the rumen microbiome. The aim of this study was to compare the microbial community composition of cow rumen and manure with respect to plant fiber-digesting microbes. Bacterial and methanogenic communities of rumen and manure samples were examined by 454 amplicon sequencing of bacterial 16S rRNA genes and mcrA genes, respectively. Rumen fluid samples were dominated by Prevotellaceae (29%), whereas Ruminococcaceae was the most abundant family in the manure samples (31%). Fibrobacteraceae (12%) and Bacteroidaceae (13%) were the second most abundant families in rumen fluid and manure, respectively. The high abundances of fiber-degrading bacteria belonging to Prevotellaceae and Fibrobacteraceae might explain the better performance of anaerobic digesters inoculated with rumen fluid. Members of the genus Methanobrevibacter were the predominant methanogens in the rumen fluid, whereas methanogenic communities of the manure samples were dominated by the candidate genus Methanoplasma. Our results suggest that inoculation or bioaugmentation with fiber-digesting rumen microbiota can enhance the anaerobic digestion of lignocellulosic biomass.

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The main aim of this study was to evaluate the effect of the source of microorganisms on the selection of hydrolytic consortia dedicated to anaerobic digestion of maize silage. The selection process was investigated based on the analysis of changes in the hydrolytic activity and the diversity of microbial communities derived from (i) a hydrolyzer of a commercial agricultural biogas plant, (ii) cattle slurry and (iii) raw sewage sludge, during a series of 10 passages. Following the selection process, the adapted consortia were thoroughly analyzed for their ability to utilize maize silage and augmentation of anaerobic digestion communities. The results of selection of the consortia showed that every subsequent passage of each consortium leads to their adaptation to degradation of maize silage, which was manifested by the increased hydrolytic activity of the adapted consortia. Biodiversity analysis (based on the 16S rDNA amplicon sequencing) confirmed the changes microbial community of each consortium, and showed that after the last (10th) passage all microbial communities were dominated by the representatives of Lactobacillaceae, Prevotellaceae, Veillonellaceae. The results of the functional analyses showed that the adapted consortia improved the efficiency of maize silage degradation, as indicated by the increase in the concentration of glucose and volatile fatty acids (VFAs), as well as the soluble chemical oxygen demand (sCOD). Moreover, bioaugmentation of anaerobic digestion communities by the adapted hydrolytic consortia increased biogas yield by 10-29%, depending on the origin of the community. The obtained results also indicate that substrate input (not community origin) was the driving force responsible for the changes in the community structure of hydrolytic consortia dedicated to anaerobic digestion.

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Lignocellulosic substrates are widely available but not easily applied in biogas production due to their poor anaerobic degradation. The effect of bioaugmentation by anaerobic hydrolytic bacteria on biogas production was determined by the biochemical methane potential assay. Microbial biomass from full scale upflow anaerobic sludge blanket reactor treating brewery wastewater was a source of active microorganisms and brewery spent grain a model lignocellulosic substrate. Ruminococcus flavefaciens 007C, Pseudobutyrivibrio xylanivorans Mz5(T), Fibrobacter succinogenes S85 and Clostridium cellulovorans as pure and mixed cultures were used to enhance the lignocellulose degradation and elevate the biogas production. P. xylanivorans Mz5(T) was the most successful in elevating methane production (+17.8%), followed by the coculture of P. xylanivorans Mz5(T) and F. succinogenes S85 (+6.9%) and the coculture of C. cellulovorans and F. succinogenes S85 (+4.9%). Changes in microbial community structure were detected by fingerprinting techniques.

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