Diversity Of Methanogens Research Articles

Effect of short-term warming and drought on the methanogenic communities in degraded peatlands in Zoige Plateau

Peatlands in Qinghai-Tibetan are degrading with climate change and human activities. Peatland degradation and climate change affect methane emissions. Methanogens are key functional microbes during methane production; however, knowledge of methanogens in degraded peatlands is lacking. Here, we investigated the effects of short-term (1 year) warming (OTC), drought (20%), and their combination on methanogens in the degraded peatlands on the Zoige Plateau of China via qPCR and clone library analysis. The results showed that Methanomicrobiales and Methanobacteriales were predominant in all the treatments. Non-metric multidimensional scaling (NMDS) and PERMANOVA analyses showed that the methanogenic community structure among the climate change treatments was not significantly different. The relative abundance of methanogen communities showed insignificant variation among the climate change treatments. The copy number and Shannon diversity of methanogens were significantly different within the climate change treatments, and drought significantly decreased the copy number of methanogens when compared to the control. The Redundancy analysis (RDA) results and correlation analysis showed that the environmental variables measured had no significant effect on methanogenic community structure and Shannon diversity. These results indicate that methanogens are insensitive to short-term climate change in degraded peatlands. This study provides insight into methane emissions from the Zoige Plateau peatlands by focusing on the possible responses of the methanogens to climate-driven changes.

Diet containing sulfur shifted hydrogen metabolism from methanogenesis to alternative sink and influenced fermentation and gut microbial ecosystem of goats

Changing diet altered fermentation products and microbial community in the gastrointestinal tracts (GIT) of goats. This study investigated the effect of substituting corn meal (CM) with corn gluten (CG), increasing dietary sulfur, shifted H2 metabolism from CH4 to H2S, and fermentation products and microbial communities in the GIT of goats. This change redirected H2 for enteric CH4 produced during microbial fermentation is loss of energy for ruminants and contributes to rising concerns of global warming. A total of 24 goats (12 per treatment) with an average age of 10 ± 0.2 months and body weight 17.5 ± 2.67 kg (mean ± SD) were used for the study. Goats randomly assigned to one of the two treatments, 400 g/kg DM of CM or CG, in a randomized block design. Goats were euthanized and GIT contents used to determine dissolved gasses, volatile fatty acids (VFA), and microbial community composition. Goats fed CG had increased isobutyrate (p < 0.05) and isovalerate molar proportions, whilst having reduced acetate to propionate ratio in both the foregut and hindgut compared with those fed CM. Goats that consumed CG had greater H2S (p < 0.01), and lower H2 and dissolved CH4 (dCH4) in the foregut and hindgut than those consuming CM. The foregut and hindgut of goats had higher (p < 0.01) 16 S rRNA gene copies of bacteria and methanogens, and 18 S rRNA gene copies of protozoa and fungi than in the cecum > small intestine. However, bacterial species were more abundant (p < 0.05) in the foregut > hindgut > small intestine. Goats fed CG had higher (p < 0.01) gene copies of microbiota and cellulolytic bacteria, whereas starch utilizing bacterial species were less in the foregut and hindgut than those fed CM. Higher (p < 0.05) methanogenic diversity and abundances of Methanimicrococcus and Methanomicrobium were observed in goats that consumed CG, whilst containing lower Methanobrevibacter populations than those receiving CM. This study suggested that using CG in place of CM, could improve feed utilization efficiency and reduce greenhouse gas emissions in ruminant production.

Microbial communities and metabolic pathways involved in reductive decolorization of an azo dye in a two-stage AD system

Multiple stage anaerobic system was found to be an effective strategy for reductive decolorization of azo dyes in the presence of sulfate. Bulk color removal (56–90%) was achieved concomitant with acidogenic activity in the 1st-stage reactor (R1), while organic matter removal (≤100%) and sulfate reduction (≤100%) occurred predominantly in the 2nd-stage reactor (R2). However, azo dye reduction mechanism and metabolic routes involved remain unclear. The involved microbial communities and conditions affecting the azo dye removal in a two-stage anaerobic digestion (AD) system were elucidated using amplicon sequencing (16S rRNA, fhs, dsrB and mcrA) and correlation analysis. Reductive decolorization was found to be co-metabolic and mainly associated with hydrogen-producing pathways. We also found evidence of the involvement of an azoreductase from Lactococcus lactis. Bacterial community in R1 was sensitive and shifted in the presence of the azo dye, while microorganisms in R2 were more protected. Higher diversity of syntrophic-acetate oxidizers, sulfate reducers and methanogens in R2 highlights the role of the 2nd-stage in organic matter and sulfate removals, and these communities might be involved in further transformations of the azo dye reduction products. The results improve our understanding on the role of different microbial communities in anaerobic treatment of azo dyes and can help in the design of better solutions for the treatment of textile effluents.

Symposium review: Understanding the role of the rumen microbiome in enteric methane mitigation and productivity in dairy cows.

Ruminants are one of the largest sources of global CH4 emissions. This enteric CH4 is exclusively produced by methanogenic archaea as a natural product during microbial fermentation in the reticulorumen. As CH4 formation leads to a gross energy loss for the ruminant host and is also an environmental issue, several CH4 mitigation approaches have been investigated, but results have been inconsistent, which may be partially attributed to a lack of understanding of the mechanistic basis of methanogenesis and the effect of inhibitors on individual methanogenic lineages and other fermenting microbes in the rumen. Methanogenic archaea are obligatory anaerobes that can reduce CO2, methanol, or methylamines or cleave acetate to form CH4. Although methanogens work toward a common goal of generating energy through the formation of CH4, individual methanogenic lineages differ in their physiological and metabolic capabilities, which can differentially affect H2 transactions and CH4 formation. Using advanced omic approaches, recent research has revealed that less abundant methanol-utilizing Methanosphaera and methylamine- and methanol-utilizing Methanomassiliicoccales lineages are positively correlated with CH4 emissions and may have a greater share in overall CH4 production compared with more abundant CO2-reducing methanogens than previously thought. These data imply that the diversity as well as the abundance of methanogens is important in CH4 formation, and that this diversity is influenced by H2 availability and interactions within and between H2-producing microbes in the rumen. These complex interactions between microbes and H2 are further influenced by variations in dietary, host, and environmental conditions. This review discusses critical knowledge gaps underlying methanogen diversity and its link to CH4 formation, formation of specific bacteria-archaeal cohorts, and how H2 production and utilization are regulated between these cohorts during normal and inhibited methanogenesis. Addressing these knowledge gaps has the potential to lead to the development of novel strategies or to complement existing strategies to effectively reduce CH4 formation while also improving productivity in dairy cows.

Shifting microbial communities perform anaerobic oxidation of methane and methanogenesis in sediments from the Shenhu area of northern south China sea during long-term incubations

Microbial communities and their associated metabolic activities, methanogenesis and anaerobic oxidation of methane (AOM), are the key components of carbon biocycles in continental margin sediments. The composition and diversity of microbial communities in a methanic environment have been widely investigated, but identifying direct correlations between microbial communities and their activities remains a challenge. Here, we investigated shifting microbial communities that performed methanogenesis and AOM in long-term incubations (the longest is up to 199 days). AOM, methanogenesis, sulfate reduction and iron reduction occurred during the incubation, and 16S rRNA gene sequencing showed that some bacteria were maintained or even enriched during the incubation compared to the environmental samples. In contrast, archaeal diversity was reduced, and only some uncultured archaea belonging to the phylum Bathyarchaeota were enriched after treatment with a high sulfate concentration (29.38 mM), suggesting that sulfate might promote their enrichment. Well-known anaerobic methanotrophic archaea (ANME) were not detected, and SEEP-SRB1, which is in syntrophy with ANME, decreased to approximately zero after the incubation. The abundance of known methanogens, such as genera Methanococcoides and Methanosarcina, increased slightly in some incubations but was still present at a low relative abundance (&amp;lt;0.15%). Incubation with a lower sulfate concentration (4 mM) and higher iron content resulted in greater δ13CO2 accumulation, indicating that iron may be the additional electron acceptor for AOM. Based on these results, other unknown or unconventional phylotypes or pathways of methanogenesis and AOM may occur during the incubation. Thus, the diversity of methanogens and anaerobic methanotrophs warrants further investigation.

Diversity of gut methanogens and functional enzymes associated with methane metabolism in smallholder dairy cattle.

Methane is a greenhouse gas with disastrous consequences when released to intolerable levels. Ruminants produce methane during gut fermentation releasing it through belching and/or flatulence. To better understand the diversity of methanogens and functional enzymes associated with methane metabolism in dairy cows, 48 samples; 6 rumen fluid and 42 dung samples were collected from Kenyan and Tanzanian farms and were analyzed using shotgun metagenomic approach. Statistical analysis for species frequency, relative abundance, percentages, and P values were undertaken using MS Excel and IBM SPSS statistics 20. The results showed archaea from 5 phyla, 9 classes, 16 orders, 25 families, 59 genera, and 87 species. Gut sites significantly contributed to the presence and distribution of various methanogens (P < 0.01). The class Methanomicrobia was abundant in the rumen samples (~ 39%) and dung (~ 44%). The most abundant (~ 17%) methanogen species identified was Methanocorpusculum labreanum. However, some taxonomic class data were unclassified (~ 6% in the rumen and ~ 4% in the dung). Five functional enzymes: Glycine/Serine hydroxymethyltransferase, Formylmethanofuran-tetrahydromethanopterin N-formyltransferase, Formate dehydrogenase, anaerobic carbon monoxide dehydrogenase, and catalase-peroxidase associated with methane metabolism were identified. KEGG functional metabolic analysis for the enzymes identified during this study was significant (P < 0.05) for five metabolism processes. The methanogen species abundances from this study in numbers/kind can be utilized exclusively or jointly as indirect selection criteria for methane mitigation. When targeting functional genes of the microbes/animal for better performance, the concern not to affect the host animal's functionality should be undertaken. Future studies should consider taxonomically categorizing unclassified species.

Methanogens and Hydrogen Sulfide Producing Bacteria Guide Distinct Gut Microbe Profiles and Irritable Bowel Syndrome Subtypes.

Irritable bowel syndrome (IBS) includes diarrhea-predominant (IBS-D) and constipation-predominant (IBS-C) subtypes. We combined breath testing and stool microbiome sequencing to identify potential microbial drivers of IBS subtypes. IBS-C and IBS-D subjects from 2 randomized controlled trials (NCT03763175 and NCT04557215) were included. Baseline breath carbon dioxide, hydrogen (H 2 ), methane (CH 4 ), and hydrogen sulfide (H 2 S) levels were measured by gas chromatography, and baseline stool microbiome composition was analyzed by 16S rRNA sequencing. Microbial metabolic pathways were analyzed using Kyoto Encyclopedia of Genes and Genomes collection databases. IBS-C subjects had higher breath CH 4 that correlated with higher gut microbial diversity and higher relative abundance (RA) of stool methanogens, predominantly Methanobrevibacter , as well as higher absolute abundance of Methanobrevibacter smithii in stool. IBS-D subjects had higher breath H 2 that correlated with lower microbial diversity and higher breath H 2 S that correlated with higher RA of H 2 S-producing bacteria, including Fusobacterium and Desulfovibrio spp. The predominant H 2 producers were different in these distinct microtypes, with higher RA of Ruminococcaceae and Christensenellaceae in IBS-C/CH 4 + (which correlated with Methanobacteriaceae RA) and higher Enterobacteriaceae RA in IBS-D. Finally, microbial metabolic pathway analysis revealed enrichment of Kyoto Encyclopedia of Genes and Genomes modules associated with methanogenesis and biosynthesis of methanogenesis cofactor F420 in IBS-C/CH 4 + subjects, whereas modules associated with H 2 S production, including sulfate reduction pathways, were enriched in IBS-D. Our findings identify distinct gut microtypes linked to breath gas patterns in IBS-C and IBS-D subjects, driven by methanogens such as M. smithii and H 2 S producers such as Fusobacterium and Desulfovibrio spp, respectively.

Methanogenic diversity changes in full-scale anaerobic digesters by co-digestion of food waste and sewage sludge

Methanogenic diversity changes in full-scale anaerobic digesters by co-digestion of food waste and sewage sludge

Occurrence of methane-oxidizing bacteria and methanogenic archaea in earth’s cave systems—A metagenomic analysis

Karst ecosystems represent up to 25% of the land surface and recent studies highlight their potential role as a sink for atmospheric methane. Despite this, there is limited knowledge of the diversity and distribution of methane-oxidizing bacteria (MOB) or methanogens in karst caves and the sub-surface environment in general. Here, we performed a survey of 14 shotgun metagenomes from cave ecosystems covering a broad set of environmental conditions, to compare the relative abundance and phylogenetic diversity of MOB and methanogens, targeting biomarker genes for methane monooxygenase (pmoA and mmoX) and methyl-coenzyme M reductase (mcrA). Taxonomic analysis of metagenomes showed 0.02–1.28% of classified reads were related to known MOB, of which Gammaproteobacterial MOB were the most abundant making up on average 70% of the surveyed caves’ MOB community. Potential for biogenic methane production in caves was also observed, with 0.008–0.39% of reads classified to methanogens and was dominated by sequences related to Methanosarcina. We have also generated a cave ecosystems protein database (CEPD) based on protein level assembly of cave metagenomes that can be used to profile genes of interest.

Impacts of cyanobacterial biomass and nitrate nitrogen on methanogens in eutrophic lakes

Methanogenesis is a key process in carbon cycling in lacustrine ecosystems. Knowledge of the methanogenic pathway is important for creating mechanistic models as well as predicting methane emissions. Due to low concentrations of methyl substrates in freshwater lakes, the proportion of methylotrophic methanogenesis is believed to be negligible in such environments. However, the high abundance of methylotrophic methanogens previously detected in Dianchi Lake suggests that methylotrophic methanogenesis may be underestimated in eutrophic lakes, whereas their influencing factors and mechanisms are not yet clear. In this study, the effects of cyanobacteria biomass (CB) or/and nitrate nitrogen on methanogenesis, especially methylotrophic pathway, in eutrophic lakes were investigated using microcosm simulation experiments combined with chemical analysis and high-throughput sequencing techniques. The results showed that either CB or nitrate nitrogen had significant effects on methane flux, the archaeal diversity and community structure of methanogens. Functional prediction, together with the result of chemical analysis, revealed that CB could promote methylotrophic methanogenesis by providing methyl organic substrates, while nitrate nitrogen increased the relative abundance of obligate methylotrophic methanogens by competitively inhibiting the other two methanogenic pathways. In eutrophic lake where both CB and nitrate present at a high concentration, methylotrophic methanogenesis could play a much more important role than previously believed.

Differences in the methanogen community between the nearshore and offshore sediments of the South Yellow Sea.

The differences in methanogen abundance and community composition were investigated between nearshore and offshore sediments in the South Yellow Sea (SYS). Shannon, Simpson, and Chao1 indices revealed a higher diversity of methanogens in the nearshore sediments than in the offshore sediments. The Mann-Whitney U test demonstrated that the relative abundance of Methanococcoides was significantly higher in the offshore sediments, while the relative abundances of Methanogenium, Methanosarcina, Methanosaeta, Methanolinea, and Methanomassiliicoccus were significantly higher in the nearshore sediments (P < 0.05). The abundance of the mcrA gene in the nearshore sediments was significantly higher than that in the offshore sediments. Furthermore, a similar vertical distribution of the methanogen and sulfate-reducing bacteria (SRB) abundances was observed in the SYS sediments, implying there is potential cooperation between these two functional microbes in this environment. Finally, total organic carbon (TOC) was significantly correlated with methanogen community composition.

Diversity of rumen microbiota using metagenome sequencing and methane yield in Indian sheep fed on straw and concentrate diet

An in vivo study aiming to investigate the rumen methanogens community structure was conducted in Mandya sheep fed on straw and concentrate diet. The ruminal fluid samples were collected and processed for unravelling the rumen microbiota and methanogens diversity. Further, the daily enteric methane emission and methane yield was also quantified using the SF6 tracer technique. Results indicated that the Bacteroidetes (∼57%) and Firmicutes (25%) were two prominent affiliates of the bacterial community. Archaea represented about 2.5% of the ruminal microbiota. Methanobacteriales affiliated methanogens were the most prevalent in sheep rumen. The study inveterate that the ruminal archaea community in sheep is composed of 9 genera and 18 species. Methanobrevibacter represented the largest genus of the archaeome, while methylotrophs genera constituted only 13% of the community. Methanobrevibacter gottschalkii was the prominent methanogen, and Methaobrevibacter ruminantium distributed at a lower frequency (∼2.5%). Among Methanomassiliicoccales, Group 12 sp. ISO4-H5 constituted the most considerable fraction (∼11%). KEGG reference pathway for methane metabolism indicated the formation of methane through hydrogenotrophic and methylotrophic pathways, whereas the acetoclastic pathway was not functional in sheep. The enteric methane emission and methane yield was 19.7 g/d and 20.8 g/kg DMI, respectively. Various species of Methanobrevibacter were differently correlated, and the distribution of hydrogenotrophic methanogens mainly explained the variability in methane yield between the individual sheep. It can be inferred from the study that the hydrogenotrophic methanogens dominate the rumen archaeal community in sheep and methylotrophic/aceticlastic methanogens represent a minor fraction of the community. Further studies are warranted for establishing the metabolic association between the prevalent hydrogenotrophs and methylotrophs to identify the key reaction for reducing methane emission.

Mutual Exclusion of Methanobrevibacter Species in the Human Gut Microbiota Facilitates Directed Cultivation of a Candidatus Methanobrevibacter Intestini Representative.

ABSTRACTMethanogenic Archaea (methanogens) are a phylogenetically diverse group of microorganisms and are considered to be the most abundant archaeal representatives in the human gut. However, the gut methanogen diversity of human populations in many global regions remains poorly investigated. Here, we report the abundance and diversity of gut methanogenic Archaea in a multi-ethnic cohort of healthy Singaporeans by using a concerted approach of metagenomic sequencing, 16S rRNA gene amplicon sequencing, and quantitative PCR. Our results indicate a mutual exclusion of Methanobrevibacter species, i.e., the highly prevalent Methanobrevibacter smithii and the less prevalent Candidatus Methanobrevibacter intestini in more than 80% of the samples when using an amplicon sequencing-based approach. Leveraging on this finding, we were able to select a fecal sample to isolate a representative strain, TLL-48-HuF1, for Candidatus Methanobrevibacter intestini. The analyzed physiological parameters of M. smithii DSM 861T and strain TLL-48-HuF1 suggest high similarity of the two species. Comparative genome analysis and the mutual exclusion of the Methanobrevibacter species indicate potentially different niche adaptation strategies in the human host, which may support the designation of Candidatus M. intestini as a novel species.IMPORTANCE Methanogens are important hydrogen consumers in the gut and are associated with differing host health. Here, we determine the prevalence and abundance of archaeal species in the guts of a multi-ethnic cohort of healthy Singapore residents. While Methanobrevibacter smithii is the most prevalent and abundant methanogen in the human gut of local subjects, the recently proposed Candidatus Methanobrevibacter intestini is the abundant methanogen in a minority of individuals that harbor them. The observed potential mutual exclusion of M. smithii and Ca. M. intestini provides further support to the proposal that the two physiologically similar strains may belong to different Methanobrevibacter species.

Increased soil moisture intensifies the impacts of forest-to-pasture conversion on methane emissions and methane-cycling communities in the Eastern Amazon

Climatic changes are altering precipitation patterns in the Amazon and may influence soil methane (CH4) fluxes due to the differential responses of methanogenic and methanotrophic microorganisms. However, it remains unclear if these climate feedbacks can amplify land-use-related impacts on the CH4 cycle. To better predict the responses of soil CH4-cycling microorganisms and emissions under altered moisture levels in the Eastern Brazilian Amazon, we performed a 30-day microcosm experiment manipulating the moisture content (original moisture; 60%, 80%, and 100% of field capacity - FC) of forest and pasture soils. Gas samples were collected periodically for gas chromatography analysis, and methanogenic archaeal and methanotrophic bacterial communities were assessed using quantitative PCR and metagenomics. Positive and negative daily CH4 fluxes were observed for forest and pasture, indicating that these soils can act as both CH4 sources and sinks. Cumulative emissions and the abundance of methanogenesis-related genes and taxonomic groups were affected by land use, moisture, and their interaction. Pasture soils at 100% FC had the highest abundance of methanogens and CH4 emissions, 22 times higher than forest soils under the same treatment. Higher ratios of methanogens to methanotrophs were found in pasture than in forest soils, even at field capacity conditions. Land use and moisture were significant factors influencing the composition of methanogenic and methanotrophic communities. The diversity and evenness of methanogens did not change throughout the experiment. In contrast, methanotrophs exhibited the highest diversity and evenness in pasture soils at 100% FC. Taken together, our results suggest that increased moisture exacerbates soil CH4 emissions and microbial responses driven by land-use change in the Amazon. This is the first report on the microbial CH4 cycle in Amazonian upland soils that combined one-month gas measurements with advanced molecular methods.

Tracking the diversity and interaction of methanogens in the energy recovery process of a full-scale wastewater treatment plant

Methanogens have been significant for the achievement of carbon neutrality in wastewater treatment plants due to their crucial roles in the anaerobic digestion of sludge. Nevertheless, the phylogenetic diversity of methanogens and their versatile metabolism have been continuously investigated, the current scientific knowledge regarding these microbes appears inadequate and requires more evaluations. This study is considered an endeavor in which functional genes sequencing was used to reveal the diversity of methanogens in the sludge process of the wastewater treatment plant. The information obtained was substantially more than that employing 16s sequencing. The methanogenic microbial resources were appropriate to sustain a self-inoculated energy recovery with a potential ability to boost methane production. A constancy was observed in 16 S rRNA gene and mcrA gene sequencing results, where the bacterial or Methanosaeta concilii dominated community of DS (digest sludge) was distinct from the inoculum sources TS (total sludge), CTS (concentrated total sludge), and HTS (hydrolysis total sludge), indicating the independent development of DS. A quantitative cross-network was constructed by coupling the absolute quantify of 16 S rRNA and mcrA sequences. The Methanobacterium petrolearium actively interacted with bacteria in the DS community rather than the dominant species (Methanosaeta concilii). Moreover, the unclassified methanogens were identified to be significantly prevalent in all communities, suggesting that unknown methanogenic taxa might be imperative in accomplishing community functions. Collectively, the findings of this research study will shed light on the comprehensive knowledge of microbial communities, especially the methanogenic microbiota. This will further enhance the exploration of the phylogenetic diversity of methanogens and their corresponding impacts in energy recovery from wastewater treatment plants.

Cultivation and biogeochemical analyses reveal insights into methanogenesis in deep subseafloor sediment at a biogenic gas hydrate site

Gas hydrates deposited in subseafloor sediments are considered to primarily consist of biogenic methane. However, little evidence for the occurrence of living methanogens in subseafloor sediments has been provided. This study investigated viable methanogen diversity, population, physiology and potential activity in hydrate-bearing sediments (1–307 m below the seafloor) from the eastern Nankai Trough. Radiotracer experiments, the quantification of coenzyme F430 and molecular sequencing analysis indicated the occurrence of potential methanogenic activity and living methanogens in the sediments and the predominance of hydrogenotrophic methanogens followed by methylotrophic methanogens. Ten isolates and nine representative culture clones of hydrogenotrophic, methylotrophic and acetoclastic methanogens were obtained from the batch incubation of sediments and accounted for 0.5–76% of the total methanogenic sequences directly recovered from each sediment. The hydrogenotrophic methanogen isolates of Methanocalculus and Methanoculleus that dominated the sediment methanogen communities produced methane at temperatures from 4 to 55 °C, with an abrupt decline in the methane production rate at temperatures above 40 °C, which is consistent with the depth profiles of potential methanogenic activity in the Nankai Trough sediments in this and previous studies. Our results reveal the previously overlooked phylogenetic and metabolic diversity of living methanogens, including methylotrophic methanogenesis.

Methanogenesis pathways of methanogens and their responses to substrates and temperature in sediments from the South Yellow Sea

Although coastal sediments are major contributors to the production of atmospheric methane, the effects of environmental conditions on methanogenesis and the community of methanogenic archaea are not well understood. Here, we investigated the methanogenesis pathways in nearshore and offshore sediments from the South Yellow Sea (SYS). Moreover, the effects of the supply of methanogenic substrates (H2/CO2, acetate, trimethylamine (TMA), and methanol) and temperature on methanogenesis and the community of methanogenic archaea were further determined. Methylotrophic, hydrogenotrophic and acetotrophic methanogenesis were found to be responsible for biogenic methane production in nearshore sediments. In the offshore sediments, methylotrophic methanogenesis was the predominant methanogenic pathway. The changes in methanogenic community structure under different substrate amendments were characterized. Lower diversities were detected in substrate-amended samples with methanogenic activity. Hydrogenotrophic Methanogenium, multitrophic Methanosarcina, methylotrophic Methanococcoide, Methanococcoide or methylotrophic Methanolobus were dominant in H2/CO2-, acetate-, TMA- and methanol-amended sediment slurries, respectively. PCoA showed that the methanogen community in H2/CO2 and acetate amendments exhibited greater differences than those in other treatments. Lower temperature (10 °C) limits hydrogenotrophic and acetoclastic methanogenesis, but methylotrophic methanogenesis is much less affected. The response of methanogen diversity to the incubation temperature varied among the different substrate-amended slurries. The multitrophic methanogen Methanosarcina became increasingly abundant in H2/CO2− and acetate-amended sediment slurries when the temperature increased from 10 to 30 °C.

Effect of olive and date palm by-products on rumen methanogenic community in Barki sheep.

Rumen methanogens prevent the accumulation of fermentation gases in the rumen and generate methane that increases global warming and represents a loss in animals' gross energy. Non-traditional feed resources such as the by-products of date palm (Phoenix dactylifera) and olive (Olea europaea) trees have received attention to be used in animal feeding. This study evaluated the impact of non-traditional feed resources including olive cake (OC), discarded dates (DD), and date palm frond (DPF) in sheep diet on rumen fermentation, diversity and relative abundance of rumen methanogens. Nine adult rams were assigned to three equal groups and fed three diets: traditional concentrates mixture (S1); non-traditional concentrate mixture (S2) based on DD and OC; and (S3) composed of the same S2 concentrate supplemented with DPF as a roughage part. The results showed that rumen pH was higher with S3 diet than the other two diets. However, the S1 diet showed the highest values of total volatile fatty acids (TVFA) and rumen ammonia. In addition, the proportions of acetic and butyric acids were increased, whereas propionic acid declined in S2 and S3 compared to the S1 diet. Rumen methanogens were dominated by Methanobrevibacter that showed a numeric decline by including DD, OC, and DPF in the animal diets. Principal component analysis (PCA) based on rumen fermentation parameters and relative abundances of methanogens genera showed three distinct clusters. Also, positive and negative correlations were revealed between methanogens genera and rumen metabolites. This study expands the knowledge regarding the effect of agricultural byproducts on rumen fermentation and the methanogenic community.

Changes in Carbohydrate Composition in Fermented Total Mixed Ration and Its Effects on in vitro Methane Production and Microbiome.

The purpose of this experiment was to investigate the changes of carbohydrate composition in fermented total mixed diet and its effects on rumen fermentation, methane production, and rumen microbiome in vitro. The concentrate-to-forage ratio of the total mixed ration (TMR) was 4:6, and TMR was ensiled with lactic acid bacteria and fibrolytic enzymes. The results showed that different TMRs had different carbohydrate compositions and subfractions, fermentation characteristics, and bacterial community diversity. After fermentation, the fermented total mixed ration (FTMR) group had lower contents of neutral detergent fiber, acid detergent fiber, starch, non-fibrous carbohydrates, and carbohydrates. In addition, lactic acid content and relative abundance of Lactobacillus in the FTMR group were higher. Compared with the TMR group, the in vitro ammonia nitrogen and total volatile fatty acid concentrations and the molar proportion of propionate and butyrate were increased in the FTMR group. However, the ruminal pH, molar proportion of acetate, and methane production were significantly decreased in the FTMR group. Notably, we found that the relative abundance of ruminal bacteria was higher in FTMR than in TMR samples, including Prevotella, Coprococcus, and Oscillospira. At the same time, we found that the diversity of methanogens in the FTMR group was lower than that in the TMR group. The relative abundance of Methanobrevibacter significantly decreased, while the relative abundances of Methanoplanus and vadinCA11 increased. The relative abundances of Entodinium and Pichia significantly decreased in the FTMR group compared with the TMR group. These results suggest that FTMR can be used as an environmentally cleaner technology in animal farming due to its ability to improve ruminal fermentation, modulate the rumen microbiome, and reduce methane emissions.

Archaeal community dynamics in biogas fermentation at various temperatures assessed by mcrA amplicon sequencing using different primer pairs.

In this study, the taxonomic and functional diversity of methanogenic archaea in two parallel 120l fermenters operated at different temperatures and fed with maize silage was estimated by mcrA metabarcoding analysis using two typical primer pairs (ML and MLA) amplifying part of the functional methyl coenzyme M reductase (mcrA) gene. The alpha diversity indices showed that the ML primer pair detected a higher Operational Taxonomic Unit (OTU) abundance compared to the MLA primer pair and methanogen diversity was significantly lower in the 60°C fermenters. The beta diversity analysis showed the methanogenic community clustered together at 50°C and 40° and was statistically different from the 60°C community. Similar, to alpha diversity, beta diversity was also significantly different between primer pairs. At all temperatures analysed, the primer pairs showed a different abundance of the different methanogenic OTUs, e.g. more OTUs relative to Methanoculleus sp. with the ML primer pair, and more OTUs corresponding to Methanobacterium sp. with the MLA primer pair. Moreover, OTUs corresponding to Methanosphaera sp. and Methanobrevibacter sp. were found only by using ML primer pair, while the MLA primer pair detected sequences corresponding to Methanothrix sp.