Methanogen Community Dynamics and Methanogenic Function Response to Solid Waste Decomposition

Graphical Abstract

Effect of Methane Inhibitors on Ruminal Microbiota During Early Life and Its Relationship With Ruminal Metabolism and Growth in Calves

The present study aimed to determine whether dietary supplementation with methanogen inhibitors during early life may lead to an imprint on the rumen microbial community and change the rumen function and performance of calves to 49-weeks of rearing. Twenty-four 4-day-old Friesian x Jersey cross calves were randomly assigned into a control and a treatment group. Treated calves were fed a combination of chloroform (CF) and 9,10-anthraquinone (AQ) in the solid diets during the first 12 weeks of rearing. Afterward, calves were grouped by treatments until week 14, and then managed as a single group on pasture. Solid diets and water were offered ad libitum. Methane measurements, and sample collections for rumen metabolite and microbial community composition were carried out at the end of weeks 2, 4, 6, 8, 10, 14, 24 and 49. Animal growth and dry matter intake (DMI) were regularly monitored over the duration of the experiment. Methane emissions decreased up to 90% whilst hydrogen emissions increased in treated compared to control calves, but only for up to 2 weeks after treatment cessation. The near complete methane inhibition did not affect calves’ DMI and growth. The acetate:propionate ratio decreased in treated compared to control calves during the first 14 weeks but was similar at weeks 24 and 49. The proportions of Methanobrevibacter and Methanosphaera decreased in treated compared to control calves during the first 14 weeks; however, at week 24 and 49 the archaea community was similar between groups. Bacterial proportions at the phylum level and the abundant bacterial genera were similar between treatment groups. In summary, methane inhibition increased hydrogen emissions, altered the methanogen community and changed the rumen metabolite profile without major effects on the bacterial community composition. This indicated that the main response of the bacterial community was not a change in composition but rather a change in metabolic pathways. Furthermore, once methane inhibition ceased the methanogen community, rumen metabolites and hydrogen emissions became similar between treatment groups, indicating that perhaps using the treatments tested in this study, it is not possible to imprint a low methane microbiota into the rumen in the solid feed of pre-weaned calves.

Modeling Root Exudate Accumulation Gradients to Estimate Net Exudation Rates by Peatland Soil Depth

Root exudates accumulate as a radial gradient around the root, yet little is known about variability at the individual root level. Vertical gradients in soil properties are hypothesized to cause greater accumulation of exudates in deeper soil through hindering diffusion, increasing sorption, and decreasing mineralization. To this end, a single root exudation model coupling concentration specific exudation and depth dependent soil properties was developed. The model was parameterized for a peatland ecosystem to explore deposition to the methanogen community. Numerical experiments indicate that exudates accumulated to a greater extent in deeper soil, albeit the effect was solute specific. Rhizosphere size for glucose doubled between the 10 and 80 cm depths, while the rhizoplane concentration was 1.23 times higher. Root influx of glucose increased from 1.431 to 1.758 nmol cm−1 hr−1, representing a recapture efficiency gain of 15.74% (i.e., 69.06% versus 84.8%). Driven by increased root influx, overall net exudation rates of select sugars and amino acids varied by a factor two. Model sensitivity analysis revealed that soil depth and root influx capability are key determinants of the rhizoplane concentration and subsequently net exudation, which determines whether effluxed compounds escape the root oxic shell and are available to the methanogen community.

Diversity of Methanogens in Animals’ Gut

Methanogens are members of anaerobe microbiota of the digestive tract of mammals, including humans. However, the sources, modes of acquisition, and dynamics of digestive tract methanogens remain poorly investigated. In this study, we aimed to expand the spectrum of animals that could be sources of methanogens for humans by exploring methanogen carriage in animals. We used real-time PCR, PCR-sequencing, and multispacer sequence typing to investigate the presence of methanogens in 407 fecal specimens collected from nine different mammalian species investigated here. While all the negative controls remained negative, we obtained by PCR-sequencing seven different species of methanogens, of which three (Methanobrevibacter smithii, Methanobrevibacter millerae and Methanomassiliicoccus luminyensis) are known to be part of the methanogens present in the human digestive tract. M. smithii was found in 24 cases, including 12/24 (50%) in pigs, 6/24 (25%) in dogs, 4/24 (16.66%) in cats, and 1/24 (4.16%) in both sheep and horses. Genotyping these 24 M. smithii revealed five different genotypes, all known in humans. Our results are fairly representative of the methanogen community present in the digestive tract of certain animals domesticated by humans, and other future studies must be done to try to cultivate methanogens here detected by molecular biology to better understand the dynamics of methanogens in animals and also the likely acquisition of methanogens in humans through direct contact with these animals or through consumption of the meat and/or milk of certain animals, in particular cows.

Response of methanogen community to elevation of cathode potentials in the presence of magnetite

ABSTRACTElectromethanogenesis refers to the process where methanogens utilize electrons derived from cathodes for the reduction of CO2 to CH4. Setting of low cathode potentials is essential for this process. In this study, we test if magnetite, an iron oxide mineral widespread in environment, can facilitate the adaption of methanogen community to the elevation of cathode potentials in electrochemical reactors. Two-chamber electrochemical reactors were constructed with inoculants obtained from a paddy field soil. We elevated cathode potentials stepwise from the initial −0.6 V vs standard hydrogen electrode (SHE) to −0.5 V and then to −0.4 V over the 120 days acclimation. Only weak current consumption and CH4 production were observed in the reactors without magnetite. But biocathodes were firmly developed and significant current consumption and CH4 production were recorded in the magnetite reactors. The robustness of electro-activity in the magnetite reactors was not affected with the elevation of cathode potentials from −0.6 V to −0.4 V. But, the current consumption and CH4 production were virtually halted in the reactors without magnetite when cathode potential was elevated to −0.4 V. Methanogens related to Methanospirillum were enriched on cathode surface of the magnetite reactors at −0.4 V, while Methanosarcina relatively dominated in the reactors without magnetite. Methanobacterium also increased in the magnetite reactors but stayed off electrodes in the culture medium at −0.4 V. Apparently, magnetite greatly facilitates the development of biocathodes, and it appears that with the aid of magnetite Methanospirillum spp. can adapt to high cathode potentials performing the efficient electromethanogenesis.IMPORTANCEConverting CO2 to CH4 through bioelectrochemistry is a promising approach for development of green energy biotechnology. This process however requires setting the low cathode potentials, which takes cost. In this study, we test if magnetite, a conductive iron mineral, can facilitate the adaption of methanogens to the elevation of cathode potentials. In the two-chamber reactors constructed using inoculants obtained from a paddy field soil, biocathodes were firmly developed in the presence of magnetite, whereas only weak electro-activity was observed in the reactors without magnetite. The elevation of cathode potentials did not affect the robustness of electro-activity in the magnetite reactors over the 120 days acclimation. Methanospirillum was identified as the key methanogens associated with cathode surface during the operation at relatively high potentials. The findings reported in this study shed a new light on the adaption of methanogen community to the elevated cathode potentials in the presence of magnetite.

An Unexpectedly Broad Thermal and Salinity-Tolerant Estuarine Methanogen Community

Moderately thermophilic (Tmax, ~55 °C) methanogens are identified after extended enrichments from temperate, tropical and low-temperature environments. However, thermophilic methanogens with higher growth temperatures (Topt ≥ 60 °C) are only reported from high-temperature environments. A microcosm-based approach was used to measure the rate of methane production and methanogen community structure over a range of temperatures and salinities in sediment from a temperate estuary. We report short-term incubations (<48 h) revealing methanogens with optimal activity reaching 70 °C in a temperate estuary sediment (in situ temperature 4–5 °C). While 30 °C enrichments amended with acetate, H2 or methanol selected for corresponding mesophilic trophic groups, at 60 °C, only hydrogenotrophs (genus Methanothermobacter) were observed. Since these methanogens are not known to be active under in situ temperatures, we conclude constant dispersal from high temperature habitats. The likely provenance of the thermophilic methanogens was studied by enrichments covering a range of temperatures and salinities. These enrichments indicated that the estuarine sediment hosted methanogens encompassing the global activity envelope of most cultured species. We suggest that estuaries are fascinating sink and source environments for microbial function study.

Stool methanogens in intestine mammal species

Methanogens are being members of anaerobe microbiota of the digestive tract of both human and mammals. However, the sources, modes of acquisition and dynamics of digestive tract methanogens remain poorly investigated. In this study, we aimed to expand the spectrum of animals which could be sources of methanogens for human, by exploring methanogen carriage in animals in contact with the general population or with some restricted populations; comparing the repertoire of animal methanogens with the one of human methanogens in order to question methanogens as zoonotic microorganisms. We used RT-PCR, PCR-sequencing and multispacer sequence typing to investigate the presence of methanogens in 407 fecal specimens collected from nine different mammalian species. We detected by RT-PCR, the presence of methanogen DNA in all mammals here investigated and none of the negative controls. We obtained by sequencing, seven different species of methanogens, of which three (Methanobrevibacter smithii, Methanobrevibacter millerae and Methanomassiliicoccus luminyensis) are known to be part of the methanogens present in the human digestive tract. We obtained 24 M. smithii by PCR-sequencing including 12/24 (50%) in pigs, 6/24 (25%) in dogs, 4/24 (16.66%) in cats, and 1/24 (4.16%) in both sheep and horses. Genotyping these 24 M. smithii revealed five different genotypes, all know in humans. Our results are fairly representative of the methanogen community present in the digestive tract of certain animals domesticated by humans and other future studies must be done to try to cultivate methanogens here detected by molecular biology to better understand the dynamics of methanogens in animals and also the likely acquisition of methanogens in humans through direct contact with these animals or through consumption of the meat and/or milk of certain animals, in particular cows.

A Membership-Fusing Model for Characterizing the Shift of Methanogen Community in a Three-Stage Sludge-Treatment Process

Anaerobic digestion (AD) is an economical and effective method to treat sludge. AD with several pretreatments is the prior process to treat surplus sludge for a wastewater treatment plant. During a sludge-treatment process, various methanogens play their specific role in each sludge-processing stage where different methanogens predominate. Therefore, an expert in the shift of methanogen community could facilitate the workers in a plant to understand the efficiency of the sludge-treatment process. In this paper, a membership-fusing model is established to characterize the shift of methanogen community in a three-stage sludge-treatment process. The introduction of fuzzy sets clarifies the vagueness of the methanogen community structure between two processing stages. Dempster–Shafer (DS) evidence theory effectively alleviates the data error generated among paralleling samples. The accuracy of the model was verified, and the result shows the model could clearly distinguish the methanogen community structure of the three stages and make accurate judgment on the processing stage affiliation. The reliability of the model in dealing with different numbers of conflict data was proved and the experiment indicates the model could make a reliable judgment on the processing stage affiliation by reasonably fusing the interference data.