Methanogenic Archaeal Populations Research Articles

The Potential of Digested Sludge-Assimilating Microflora for Biogas Production from Food Processing Wastes.

Food processing wastes (FPWs) are residues generated in food manufacturing, and their composition varies depending on the type of food product being manufactured. Therefore, selecting and acclimatizing seed microflora during the initiation of biogas production is crucial for optimal outcomes. The present study examined the biogas production capabilities of digested sludge-assimilating and biogas-yielding soil (DABYS) and enteric (DABYE) microflorae when used as seed cultures for biogas production from FPWs. After subculturing and feeding these microbial seeds with various FPWs, we assessed their biogas-producing abilities. The subcultures produced biogas from many FPWs, except orange peel, suggesting that the heterogeneity of the bacterial members in the seed microflora facilitates quick adaptation to FPWs. Microflorae fed with animal-derived FPWs contained several methanogenic archaeal families and produced methane. In contrast, microflorae fed with vegetable-, fruit-, and crop-derived FPWs generated hydrogen, and methanogenic archaeal populations were diminished by repeated subculturing. The subcultured microflorae appear to hydrolyze carbohydrates and protein in FPWs using cellulase, pectinase, or protease. Despite needing enhancements in biogas yield for future industrial scale-up, the DABYS and DABYE microflorae demonstrate robust adaptability to various FPWs.

Biological pretreatment with Trametes versicolor to enhance methane production from lignocellulosic biomass: A metagenomic approach

The presence of poorly biodegradable components in lignocellulosic biomass limits the methane recovery in anaerobic digesters. The main reason to go for aerobic pretreatment before anaerobic digestion (AD) is to enable enzymatic cleavage of the aromatic rings in lignin by oxygen since it cannot be efficiently degraded under anaerobic conditions. In this study, the advantage of highly-cellulolytic white-rot fungi Trametes versicolor was taken by aerobic pretreatment prior to anaerobic co-digestion of cow manure and selected cereal crop materials (i.e. wheat, rye, barley, triticale) harvested at different stages. Fungal pretreatment improved the methane yield by 10%–18% and cellulose degradation up to 80%. Furthermore, higher volatile fatty acid (VFA) speciation was found in the anaerobic digesters upon fungal pretreatment. 16S rRNA gene amplicon sequencing revealed a more diverse microbial community in the fungal-pretreated anaerobic digesters. Generally, typically-detected bacterial species dominated the digesters; except that Synergistetes was only enriched in the fungal-pretreated digesters. Although Methanosarcianease was the predominant methanogenic archaea, a more diverse methanogenic population was identified in the fungal-pretreated digesters in which Methanobacteriaceaa and Methanomibrobiaceae also took role during biomethanation. Comparatively more unique microbiome of biogas reactors upon fungal pretreatment synergistically affected VFA production, cellulose degradation and eventually methane yield in an affirmative way. Considering the functional importance of bacterial and methanogenic archaeal populations, elevated knowledge of the microbial structures is essential for minimizing process failures and for creating strategies for process optimization of lignocellulose based-AD.

Effects of transgenic Bt rice on the active rhizospheric methanogenic archaeal community as revealed by DNA-based stable isotope probing.

This study aimed to investigate the influence of planting Cry1Ab/Cry1Ac gene expressing rice (Bt rice) on rhizospheric active methanogenic archaeal communities. The nontransgenic parental line was used as the control (Ck rice). DNA-based stable isotope probing (DNA-SIP) technology traced the rhizospheric active methanogens at the tillering stage. The results revealed significantly lower CH4 emission flux from Bt soil than that from Ck soil during the whole growth period. The active methanogenic community composition remained stable. The RC-I lineage (77·9-79·8%) and Methanosaetaceae (13·9-15·1%) were the predominant active methanogens in Bt and Ck rice rhizospheres. However, the abundance of functionally active methanogens in the Bt rice rhizosphere was significantly reduced. Lower levels of root exudates (that included carbohydrate and organic acids) from Bt rice were also detected at the tillering stage. This study found that the genetic modification of rice reduced the potential methanogenic substrates came from plant-derived root exudates, which represented an important factor in reducing CH4 generation and active methanogenic archaeal abundance in Bt rhizosphere soil. The effect of genetically modified (GM) insect-resistant crops on soil micro-organisms has become an issue of public concern, especially the indirect effect of plant metabolisms caused by the insertion of foreign genes. Methanogenesis, which is regarded as a critical ecological process in paddy soil, is influenced by plant root exudates; these are mainly derived from photosynthesis. The variations in root exudates across the Bt and Ck rice suggested the indirect influence of foreign gene insertion. DNA-SIP successfully traced the active methanogenic archaeal populations assimilating 13 C-labelled photosynthetic carbon and found a strong influence of planting Bt rice on active methanogens. As a consequence, we proposed that analysis of functionally active micro-organisms is more suitable for monitoring and predicting the environmental influence of GM plants.

Effect of paddy-upland rotation on methanogenic archaeal community structure in paddy field soil.

Methanogenic archaea are strict anaerobes and demand highly reduced conditions to produce methane in paddy field soil. However, methanogenic archaea survive well under upland and aerated conditions in paddy fields and exhibit stable community. In the present study, methanogenic archaeal community was investigated in fields where paddy rice (Oryza sativa L.) under flooded conditions was rotated with soybean (Glycine max [L.] Merr.) under upland conditions at different rotation histories, by polymerase chain reaction (PCR)-denaturing gradient gel electrophoresis (DGGE) and real-time quantitative PCR methods targeting 16S rRNA and mcrA genes, respectively. Soil samples collected from the fields before flooding or seeding, during crop cultivation and after harvest of crops were analyzed. The abundance of the methanogenic archaeal populations decreased to about one-tenth in the rotational plots than in the consecutive paddy (control) plots. The composition of the methanogenic archaeal community also changed. Most members of the methanogenic archaea consisting of the orders Methanosarcinales, Methanocellales, Methanomicrobiales, and Methanobacteriales existed autochthonously in both the control and rotational plots, while some were strongly affected in the rotational plots, with fatal effect to some members belonging to the Methanosarcinales. This study revealed that the upland conversion for one or longer than 1 year in the rotational system affected the methanogenic archaeal community structure and was fatal to some members of methanogenic archaea in paddy field soil.

Gastrointestinal Bacterial and Methanogenic Archaea Diversity Dynamics Associated with Condensed Tannin-Containing Pine Bark Diet in Goats Using 16S rDNA Amplicon Pyrosequencing

Eighteen Kiko-cross meat goats (n = 6) were used to collect gastrointestinal (GI) bacteria and methanogenic archaea for diversity measures when fed condensed tannin-containing pine bark (PB). Three dietary treatments were tested: control diet (0% PB and 30% wheat straw (WS); 0.17% condensed tannins (CT) dry matter (DM)); 15% PB and 15% WS (1.6% CT DM), and 30% PB and 0% WS (3.2% CT DM). A 16S rDNA bacterial tag-encoded FLX amplicon pyrosequencing technique was used to characterize and elucidate changes in GI bacteria and methanogenic archaea diversity among the diets. Proteobacteria was the most dominant phylum in goats with mean relative abundance values ranging from 39.7 (30% PB) to 46.5% (control) and 47.1% (15% PB). Other phyla individually accounted for fewer than 25% of the relative abundance observed. Predominant methanogens were Methanobrevibacter (75, 72, and 49%), Methanosphaera (3.3, 2.3, and 3.4%), and Methanobacteriaceae (1.2, 0.6, and 0.7%) population in control, 15, and 30% PB, respectively. Among methanogens, Methanobrevibacter was linearly decreased (P = 0.05) with increasing PB supplementation. These results indicate that feeding PB selectively altered bacteria and methanogenic archaeal populations in the GI tract of goats.

Fluorescence in situ hybridization probing of protozoal Entodinium spp. and their methanogenic colonizers in the rumen of cattle fed alfalfa hay or triticale straw

To develop and test a fluorescence in situ hybridization (FISH) based technique and to identify and quantify simultaneously those methanogenic populations colonizing Entodinium spp. in the rumen of cows fed different forages. New FISH probes targeting protozoal Entodinium spp. were designed and used together with FISH probes for methanogens in the cow rumen. The composition and relative abundance of methanogenic populations colonizing Entodinium simplex-, E. caudaum- and Entodinium furca-related populations were similar. Methanogens including Methanobrevibacter thaueri, Methanobrevibacter millerae and Methanobrevibacter smithii, and members of Methanomicrobium and Methanosphaera were generally the predominant colonizers of protozoa, regardless of the forage fed to cattle. Individual animals appeared to differ in which ruminal methanogenic populations colonized each of the individual Entodinium spp. Simultaneous FISH probing is shown here to be a reliable and effective approach to investigate the dynamics of symbiotic relationships between ruminal protozoa and methanogens at a single cell level. Phylogenetically closely related Entodinium spp. were colonized by similar methanogenic populations regardless of the forage fed. This is the first report of the methanogenic archaeal populations that specifically colonize Entodinium spp. as identified using simultaneous FISH probing.

Molecular analysis of methanogenic archaea in the forestomach of the alpaca (Vicugna pacos)

BackgroundMethanogens that populate the gastrointestinal tract of livestock ruminants contribute significantly to methane emissions from the agriculture industry. There is a great need to analyze archaeal microbiomes from a broad range of host species in order to establish causal relationships between the structure of methanogen communities and their potential for methane emission. In this report, we present an investigation of methanogenic archaeal populations in the foregut of alpacas.ResultsWe constructed individual 16S rRNA gene clone libraries from five sampled animals and recovered a total of 947 sequences which were assigned to 51 species-level OTUs. Individuals were found to each have between 21 and 27 OTUs, of which two to six OTUs were unique. As reported in other host species, Methanobrevibacter was the dominant genus in the alpaca, representing 88.3% of clones. However, the alpaca archaeal microbiome was different from other reported host species, as clones showing species-level identity to Methanobrevibacter millerae were the most abundant.ConclusionFrom our analysis, we propose a model to describe the population structure of Methanobrevibacter-related methanogens in the alpaca and in previously reported host species, which may contribute in unraveling the complexity of symbiotic archaeal communities in herbivores.

Composition of Archaeal Community in a Paddy Field as Affected by Rice Cultivar and N Fertilizer

Methanogenesis in paddy fields is significantly influenced by environmental and field management factors such as rice cultivar and nitrogenous fertilizer. However, it has been unclear whether such effects are reflected in the structure of methanogenic archaeal populations. In the present study, molecular analyses including cloning and sequencing and terminal restriction fragment length polymorphism (T-RFLP) fingerprinting of archaeal 16S rRNA genes were used to characterize the methanogenic archaeal assemblages and to identify the effect of environmental variables including rice cultivar and N fertilizer on archaeal community compositions in a Chinese paddy field soil. The correlation between methanogenic archaeal composition and environmental variables was explored by correspondence analysis. The results showed that the spatial or niche factor (rice roots versus rhizosphere, surface, and the deeper layer soils) had the greatest influence on the archaeal community composition. There was an obvious enrichment or selection of hydrogenotrophic as opposed to acetoclastic methanogens by rice roots. The archaeal community also changed, though slightly, between the rhizosphere and bulk soils and between the surface soil and the deeper layer soil. However, rice cultivar and N fertilizer appear to have an effect only on methanogens tightly associated with rice roots.

Dynamics of sulfidogenesis associated to methanogenesis in horizontal-flow anaerobic immobilized biomass reactor

Two bench-scale horizontal anaerobic fixed bed reactors were tested to remove both sulfate and organic matter from wastewater. First, the reactors (R1 and R2) were supplied with synthetic wastewater containing sulfate and a solution of ethanol and volatile fatty acids. Subsequently, R1 and R2 were fed with only ethanol or acetate, respectively. The substitution to ethanol in R1 increased the sulfate reduction efficiency from 83% to nearly 100% for a chemical oxygen demand to sulfate (COD/sulfate) ratio of 3.0. In contrast, in R2, the switch in carbon source to acetate strongly decreased sulfidogenesis and the maximum sulfate reduction achieved was 47%. Process stability in long-term experiments and high removal efficiencies of both organic matter and sulfate were achieved with ethanol as the sole carbon source. The results allow concluding that syntrophism instead of competition between the sulfate reducing bacteria and acetoclastic methanogenic archaeal populations prevailed in the reactor.

Analysis of the Sulfate-Reducing Bacterial and Methanogenic Archaeal Populations in Contrasting Antarctic Sediments

Analysis of the Sulfate-Reducing Bacterial and Methanogenic Archaeal Populations in Contrasting Antarctic Sediments

Analysis of the sulfate-reducing bacterial and methanogenic archaeal populations in contrasting Antarctic sediments.

The distribution and activity of communities of sulfate-reducing bacteria (SRB) and methanogenic archaea in two contrasting Antarctic sediments were investigated. Methanogenesis dominated in freshwater Lake Heywood, while sulfate reduction dominated in marine Shallow Bay. Slurry experiments indicated that 90% of the methanogenesis in Lake Heywood was acetoclastic. This finding was supported by the limited diversity of clones detected in a Lake Heywood archaeal clone library, in which most clones were closely related to the obligate acetate-utilizing Methanosaeta concilii. The Shallow Bay archaeal clone library contained clones related to the C(1)-utilizing Methanolobus and Methanococcoides and the H(2)-utilizing Methanogenium: Oligonucleotide probing of RNA extracted directly from sediment indicated that archaea represented 34% of the total prokaryotic signal in Lake Heywood and that Methanosaeta was a major component (13.2%) of this signal. Archaea represented only 0.2% of the total prokaryotic signal in RNA extracted from Shallow Bay sediments. In the Shallow Bay bacterial clone library, 10.3% of the clones were SRB-like, related to Desulfotalea/Desulforhopalus, Desulfofaba, Desulfosarcina, and Desulfobacter as well as to the sulfur and metal oxidizers comprising the Desulfuromonas cluster. Oligonucleotide probes for specific SRB clusters indicated that SRB represented 14.7% of the total prokaryotic signal, with Desulfotalea/Desulforhopalus being the dominant SRB group (10.7% of the total prokaryotic signal) in the Shallow Bay sediments; these results support previous results obtained for Arctic sediments. Methanosaeta and Desulfotalea/Desulforhopalus appear to be important in Lake Heywood and Shallow Bay, respectively, and may be globally important in permanently low-temperature sediments.

In situ measurement of archaeal and bacterial specific growth rates in two stage high rate anaerobic treatment systems

The thymidine assay is applied and evaluated as a direct measure of archaeal and bacterial volumetric growth rate in the first and second stages of a full scale High Rate Anaerobic Treatment system (HRAT). By coupling the method to an epifluorescent microscopic technique we show how to measure microbial specific growth rates in situ. In the first stage reactor, the acidogenic (hydrolytic-fermentative) apparent bacterial specific growth rate measurements (2.2 d –1 ) agreed with that predicted by the hydraulic residence time. For the methanogenic archaeal populations in the second stage upflow anaerobic sludge blanket (UASB) reactor the measurement was the same as published values. The results also showed that in the UASB reactor the archaeal growth dynamics in the liquid phase and 'sludge' granules were the same.