Search Results (14)

The aim of this experiment is to explore the effect of sodium sulfate (Na₂SO₄) on methane reduction in the rumen, and its impact on anaerobic methane-oxidizing archaea (ANME). Using mixed rumen fluid from four Angus cattle fistulas, this study conducted an in vitro fermentation. Adding Na₂SO₄ to the fermentation substrate resulted in sulfur concentrations in the substrate of 0.4%, 0.6%, 0.8%, 1.0%, 1.2%, 1.4%, 1.6%, 1.8%, 2.0%, 2.2%, and 2.4%. The gas production rate and methane yield were measured using an in vitro gas production method. Subsequently, the fermentation fluid was collected to determine the fermentation parameters. The presence of ANME in the fermentation broth, as well as the relationship between the number of bacteria, archaea, sulfate reducing bacteria (SRB), ANME, and the amount of Na₂SO₄ added to the substrate, were measured using qPCR. The results showed that: (1) the addition of Na₂SO₄ could significantly reduce CH₄ production and was negatively correlated with CO₂ production; (2) ANME-1 and ANME-2c did exist in the fermentation broth; (3) the total number of archaea, SRB, ANME-1, and ANME-2c increased with the elevation of Na₂SO₄. The above results indicated that Na₂SO₄ could mitigate methane production via sulfate-dependent anaerobic methane oxidation (S-DAMO) in the rumen. In the future management of beef cattle, including sodium sulfate in their diet can stimulate S-DAMO activity, thereby promoting a reduction in methane emissions. Full article

Anaerobic digestion of waste-activated sludge (WAS) towards biogas recovery is constrained by the limited hydrolysis and inhibited acetogenesis steps that hinder subsequent energy recovery. This study employed Fe(VI)/S(IV) oxidation to enhance the WAS solubilization and coupled it with the syntrophic interaction of hydrogen-producing acetogen (HPA) and sulfate-reducing bacteria (SRB) to stimulate the successive procedure towards methane production. Results showed that the dosage ratio of HPA-SRB to WAS (H-S-W) with 1:1:50 outperformed with the highest methane production potential (11.63 ± 1.87 mL CH₄/(g VSS·d). Meanwhile, the efficient and sequential process from acetogenesis to methanogenesis stimulated by HPA-SRB was evidenced by a significant decrease of 30.2% in the acetate concentration. The microbial community structure further manifested the crucial role of HPA-SRB with increased abundance of Desulfobulbus (2.07%), Syntrophomonas (1.24%) and Smithella (1.63%), which stimulated acetophilic methanogen boost with Methanobacterium dominating with 77.51% in H-S-W₁₀₀. Furthermore, the positive syntrophic relationships among HPA-SRB and acetophilic methanogens towards methane production were confirmed via molecular ecological network and canonical correspondence analysis. This study highlighted the syntrophic cooperation of the mixed consortia of HPA and SRB on methane production based on Fe(VI)/S(IV) pretreatment and provided the theoretical and technical basis for the potential implementation of novel methanogenesis technology for WAS treatment. Full article

Time-of-Flight–Secondary Ion Mass Spectrometry (ToF-SIMS) using a bismuth liquid metal ion source was utilized to characterize and image microbial biomarkers within dolomite from early-middle Miocene coastal mud volcano outcrops in Kuwait. ToF-SIMS analysis revealed biomarkers of ancient microbial consortia of sulfate reducers and methane oxidizers participating in the anaerobic oxidation of methane. The identified lipid biomarkers comprised 17α(H),21β(H)-Norhopane, Hop-17(21)-ene or Hop-22(29)-ene (diploptene), non-isoprenoidal dialkyl glycerol diethers (DAGEs), and Diacylglycerol esters (DGs). The ion µ-scale images of carbonate rocks showed two characteristic styles: (1) high signal intensity of dolomite, halite, and biomarkers, where the biomarkers demonstrate a distinctive co-localization pattern with both dolomite and halite; and (2) a lack of dolomite, halite low signal intensity, and an absence of biomarker co-localization patterns. Our results highlight three remarkable observations. Firstly, the concomitance of dolomite and halite suggests a common source of magnesium and sodium supply, likely from the hypersaline Al-Subiya sabkha. This emphasizes the importance of hypersaline seawater for dolomite formation. Secondly, microbial biomarkers correspond to methane- and sulfate-rich conditions under which dolomite was formed. Lastly, the high intensity of biomarker signals and their association with dolomite and halite indicate that the consortia involved in dolomite formation have preferences for high-salinity conditions. The three observations align with previous studies that have highlighted the importance of methane-sulfate redox, high salinity, and halophilic microbes for dolomite formation. This work is the first to acknowledge ancient microbial biomarkers within Miocene dolomite in Kuwait, which aims to broaden the understanding of the biogeochemical processes triggering dolomite formation in similar environments and ancient geologic settings. Full article

In the context of global warming and carbon neutrality, reducing greenhouse gas emissions is fundamental to achieving sustainable development. As an important greenhouse gas, methane has a much stronger warming effect than CO₂, and studies have demonstrated that anaerobic oxidation of methane (AOM) is important for global methane emissions. This paper systematically investigated the AOM potential and microbial community response to the input of SO₄²⁻ in the three typical salt marsh soils of the Yellow River Delta: Reed, Suaeda salsa, and Tamarisk, using SO₄²⁻ as the electron acceptor and a combination of indoor anaerobic culture and high-throughput sequencing. The results showed that after adding an appropriate concentration of SO₄²⁻, the AOM potential was significantly promoted in Tamarix soil (p < 0.05) and significantly inhibited in Reed and Suaeda salsa soil (p < 0.05); soil AOM potential and SO₄²⁻ input concentration and background values were correlated. At the microbial level, SO₄²⁻ input affected the abundance of some microorganisms. At the phylum level, the relative abundance of Proteobacteria was increased in Suaeda salsa soil, decreased in Tamarisk soil, and did not change significantly in Reed soil; that of Crenarchaeota and Desulfobacterota was significantly increased in Tamarisk soil. At the genus level, Methylophaga, Methylotenera, and Methylomonaceae became the dominant populations, and it can be inferred that these bacteria are involved in the anaerobic oxidation of methane after the input of SO₄²⁻. This study will be of great significance to the mechanistic study of AOM and the conservation of microbial diversity in the Yellow River Delta Coastal Wetland, as well as provide a scientific basis for CH₄ reduction in coastal wetlands. Full article

Bottom sediments at methane discharge sites of the Laptev Sea shelf were investigated. The rates of microbial methanogenesis and methane oxidation were measured, and the communities responsible for these processes were analyzed. Methane content in the sediments varied from 0.9 to 37 µmol CH₄ dm⁻³. Methane carbon isotopic composition (δ¹³C-CH₄) varied from −98.9 to −77.6‰, indicating its biogenic origin. The rates of hydrogenotrophic methanogenesis were low (0.4–5.0 nmol dm⁻³ day⁻¹). Methane oxidation rates varied from 0.4 to 1.2 µmol dm⁻³ day⁻¹ at the seep stations. Four lineages of anaerobic methanotrophic archaea (ANME) (1, 2a–2b, 2c, and 3) were found in the deeper sediments at the seep stations along with sulfate-reducing Desulfobacteriota. The ANME-2a-2b clade was predominant among ANME. Aerobic ammonium-oxidizing Crenarchaeota (family Nitrosopumilaceae) predominated in the upper sediments along with heterotrophic Actinobacteriota and Bacteroidota, and mehtanotrophs of the classes Alphaproteobacteria (Methyloceanibacter) and Gammaproteobacteria (families Methylophilaceae and Methylomonadaceae). Members of the genera Sulfurovum and Sulfurimonas occurred in the sediments of the seep stations. Mehtanotrophs of the classes Alphaproteobacteria (Methyloceanibacter) and Gammaproteobacteria (families Methylophilaceae and Methylomonadaceae) occurred in the sediments of all stations. The microbial community composition was similar to that of methane seep sediments from geographically remote areas of the global ocean. Full article

The treatment of livestock manure caused by the expansion of the breeding industry in China has attracted wide attention. Heavy metals in pig manure can pollute soil and water and even transfer to crops, posing harm to humans through the food chain. In this study, corn straw was selected as the additive and introduced into the anaerobic digestion. Sepiolite (SE), ferric oxide (Fe₂O₃), attapulgite (AT) and ferric sulfate (FeSO₄) were used as passivators to compare the effects of these inorganic passivators on gas production and passivation of heavy metals during the process of the anaerobic digestion. When the dry mass ratio of pig manure to straw is 8:2, the gas production efficiency is optimal. SE, AT and ferric sulfate have a much stronger ability to improve gas production performance than Fe₂O₃. The total gas production increased by 10.34%, 6.62% and 4.56%, and the average methane production concentration increased by 0.7%, 0.3% and 0.4%, respectively. The influence of SE, AT and ferric sulfate on the passivation of heavy metals is much better than Fe₂O₃, and the fractions in biological effective forms of Cu and Zn reduced by 41.87 and 19.32%, respectively. The anaerobic digestion of mixed materials is conducive to the gas production and the passivation of heavy metals. Therefore, SE, AT and ferric sulfate are selected as composite passivators, and the optimal ratio of inorganic composite passivators i: AT 7.5 g/L, ferric sulfate 5 g/L and SE 7.5 g/L, according to the results of orthogonal experiments. This study can provide a theoretical basis for the safe application of biogas fertilizers. Full article

The tanning process generates a saline effluent with high residual organics, sulfate and sulfide concentrations. The transition from a linear to circular economy requires reimagining of waste streams as potential resources. The organics in tannery effluent have the potential to be converted to renewable energy in the form of biogas if inhibitors to anaerobic digestion are removed. Hybrid linear flow channel reactors inoculated with culture-enriched halophilic sulfate reducing bacteria from saline environments were evaluated as a novel pretreatment step prior to anaerobic digestion for the concurrent removal of sulfur species and resource recovery (elemental sulfur and biogas). During continuous operation of a 4-day hydraulic retention time, the reactors were capable of near-complete sulfide oxidation (>97%) and a sulfate reduction efficiency of 60–80% with the formation of a floating sulfur biofilm containing elemental sulfur. Batch anaerobic digestion tests showed no activity on untreated tannery effluent, while the pretreated effluent yielded 130 mL methane per gram COD consumed. Full article

A combination of physicochemical and radiotracer analysis, high-throughput sequencing of the 16S rRNA, and particulate methane monooxygenase subunit A (pmoA) genes was used to link a microbial community profile with methane, sulfur, and nitrogen cycling processes. The objects of study were surface sediments sampled at five stations in the northern part of the Barents Sea. The methane content in the upper layers (0–5 cm) ranged from 0.2 to 2.4 µM and increased with depth (16–19 cm) to 9.5 µM. The rate of methane oxidation in the oxic upper layers varied from 2 to 23 nmol CH₄ L⁻¹ day⁻¹ and decreased to 0.3 nmol L⁻¹ day⁻¹ in the anoxic zone at a depth of 16–19 cm. Sulfate reduction rates were much higher, from 0.3 to 2.8 µmol L⁻¹ day⁻¹. In the surface sediments, ammonia-oxidizing Nitrosopumilaceae were abundant; the subsequent oxidation of nitrite to nitrate can be carried out by Nitrospira sp. Aerobic methane oxidation could be performed by uncultured deep-sea cluster 3 of gamma-proteobacterial methanotrophs. Undetectable low levels of methanogenesis were consistent with a near complete absence of methanogens. Anaerobic methane oxidation in the deeper sediments was likely performed by ANME-2a-2b and ANME-2c archaea in consortium with sulfate-reducing Desulfobacterota. Sulfide can be oxidized by nitrate-reducing Sulfurovum sp. Thus, the sulfur cycle was linked with the anaerobic oxidation of methane and the nitrogen cycle, which included the oxidation of ammonium to nitrate in the oxic zone and denitrification coupled to the oxidation of sulfide in the deeper sediments. Methane concentrations and rates of microbial biogeochemical processes in sediments in the northern part of the Barents Sea were noticeably higher than in oligotrophic areas of the Arctic Ocean, indicating that an increase in methane concentration significantly activates microbial processes. Full article

Terrestrial mud volcanoes (TMVs) are important natural sources of methane emission. The microorganisms inhabiting these environments remain largely unknown. We studied the phylogenetic composition and metabolic potential of the prokaryotic communities of TMVs located in the Taman Peninsula, Russia, using a metagenomic approach. One of the examined sites harbored a unique community with a high abundance of anaerobic methane-oxidizing archaea belonging to ANME-3 group (39% of all 16S rRNA gene reads). The high number of ANME-3 archaea was confirmed by qPCR, while the process of anaerobic methane oxidation was demonstrated by radioisotopic experiments. We recovered metagenome-assembled genomes (MAGs) of archaeal and bacterial community members and analyzed their metabolic capabilities. The ANME-3 MAG contained a complete set of genes for methanogenesis as well as of ribosomal RNA and did not encode proteins involved in dissimilatory nitrate or sulfate reduction. The presence of multiheme c-type cytochromes suggests that ANME-3 can couple methane oxidation with the reduction of metal oxides or with the interspecies electron transfer to a bacterial partner. The bacterial members of the community were mainly represented by autotrophic, nitrate-reducing, sulfur-oxidizing bacteria, as well as by fermentative microorganisms. This study extends the current knowledge of the phylogenetic and metabolic diversity of prokaryotes in TMVs and provides a first insight into the genomic features of ANME-3 archaea. Full article

(1) Background: The geothermal spring of La Crouen (New Caledonia) discharges warm (42 °C) alkaline water (pH~9) enriched in dissolved nitrogen with traces of methane, but its microbial diversity has not yet been studied. (2) Methods: Cultivation-dependent and -independent methods (e.g., Illumina sequencing and quantitative PCR based on 16S rRNA gene) were used to describe the prokaryotic diversity of this spring. (3) Results: Prokaryotes were mainly represented by Proteobacteria (57% on average), followed by Cyanobacteria, Chlorofexi, and Candidatus Gracilibacteria (GN02/BD1-5) (each > 5%). Both potential aerobes and anaerobes, as well as mesophilic and thermophilic microorganisms, were identified. Some of them had previously been detected in continental hyperalkaline springs found in serpentinizing environments (The Cedars, Samail, Voltri, and Zambales ophiolites). Gammaproteobacteria, Ca. Gracilibacteria and Thermotogae were significantly more abundant in spring water than in sediments. Potential chemolithotrophs mainly included beta- and gammaproteobacterial genera of sulfate-reducers (Ca. Desulfobacillus), methylotrophs (Methyloversatilis), sulfur-oxidizers (Thiofaba, Thiovirga), or hydrogen-oxidizers (Hydrogenophaga). Methanogens (Methanobacteriales and Methanosarcinales) were the dominant Archaea, as found in serpentinization-driven and deep subsurface ecosystems. A novel alkaliphilic hydrogenotrophic methanogen (strain CAN) belonging to the genus Methanobacterium was isolated, suggesting that hydrogenotrophic methanogenesis occurs at La Crouen. Full article

Mud volcanoes are prominent geological structures where fluids and gases from the deep subsurface are discharged along a fracture network in tectonically active regions. Microbial communities responsible for sulfur and methane cycling and organic transformation in terrestrial mud volcanoes remain poorly characterized. Using a metagenomics approach, we analyzed the microbial community of bubbling fluids retrieved from an active mud volcano in eastern Crimea. The microbial community was dominated by chemolithoautotrophic Campylobacterota and Gammaproteobacteria, which are capable of sulfur oxidation coupled to aerobic and anaerobic respiration. Methane oxidation could be enabled by aerobic Methylococcales bacteria and anaerobic methanotrophic archaea (ANME), while methanogens were nearly absent. The ANME community was dominated by a novel species of Ca. Methanoperedenaceae that lacked nitrate reductase and probably couple methane oxidation to the reduction of metal oxides. Analysis of two Ca. Bathyarchaeota genomes revealed the lack of mcr genes and predicted that they could grow on fatty acids, sugars, and proteinaceous substrates performing fermentation. Thermophilic sulfate reducers indigenous to the deep subsurface, Thermodesulfovibrionales (Nitrospirae) and Ca. Desulforudis (Firmicutes), were found in minor amounts. Overall, the results obtained suggest that reduced compounds delivered from the deep subsurface support the development of autotrophic microorganisms using various electron acceptors for respiration. Full article

Marine anaerobic methane oxidation (AOM) is generally assumed to be coupled to sulfate reduction, via a consortium of anaerobic methane-oxidizing archaea (ANME) and sulfate-reducing bacteria (SRB). ANME-1 are, however, often found as single cells, or only loosely aggregated with SRB, suggesting they perform a form of AOM independent of sulfate reduction. Oxidized metals and humic substances have been suggested as potential electron acceptors for ANME, but up to now, AOM linked to reduction of these compounds has only been shown for the ANME-2 and ANME-3 clades. Here, the effect of the electron acceptors anthraquinone-disulfonate (AQDS), a humic acids analog, and Fe³⁺ on anaerobic methane oxidation were assessed by incubation experiments with anoxic Black Sea water containing ANME-1b. Incubation experiments with ¹³C-methane and AQDS showed a stimulating effect of AQDS on methane oxidation. Fe³⁺ enhanced the ANME-1b abundance but did not substantially increase methane oxidation. Sodium molybdate, which was added as an inhibitor of sulfate reduction, surprisingly enhanced methane oxidation, possibly related to the dominant abundance of Sulfurospirillum in those incubations. The presented data suggest the potential involvement of ANME-1b in AQDS-enhanced anaerobic methane oxidation, possibly via electron shuttling to AQDS or via interaction with other members of the microbial community. Full article

Sediments at marine cold seep areas provide potential archives of past fluid flow, which allow insights into the evolution of past methane seepage activities. However, signals for anaerobic oxidation of methane (AOM) might be obscured in bulk sediments in cold-seep settings due to several factors, especially flood and turbidite deposition. Comprehensive inorganic data were gathered in this study to explore the availability of related records at cold seeps and to provide insights into the evolution of past methane seepage activities. Sediments collected from the site 973-4 in the Taixinan Basin on the northern slope of the South China Sea were characterized in terms of total carbon and sulfur, δ¹³C values of total organic carbon (δ¹³C_TIC), δ³⁴S values of chromium reducible sulfur (δ³⁴S_CRS), and foraminiferal oxygen and carbon isotopes. The results confirmed a strong correlation between formation of authigenic minerals and AOM. Moreover, the ³⁴S enrichments and abundant chromium reducible sulfur (CRS) contents in the authigenic sulfides in the sulfate–methane transition zone (SMTZ) within 619–900 cm below seafloor (cmbsf) reflected past high methane fluxes supported by constant methane seepages. Lithological distribution and AMS (Accelerator Mass Spectra) ¹⁴C dating of planktonic foraminifera show that the turbidite (~35.14 ka) was related to a foraminifera-rich interval (Unit II: 440-619 cmbsf) and increased carbonate productivity during the last glacial maximum (LGM). Enrichment of Mo and U was observed accompanied by low contents of nutrient metals (Al, Ti, V, Ni, Fe, Mn, and Cu) in Unit II. The foraminifera-rich interval (Unit II) of cold seep sediments was probably linked to the phenomenon of inconsecutive sedimentary sequence due to the turbidites, which resulted in the lack of Fe, Mn, and Ba enrichment. There is no U enrichment but only Mo enrichment within Unit III, which might be related to H₂S produced by AOM during the methane seepages. Based on the above results, it can be speculated that this area has experienced multiple-episodes of methane seep events. Further exploration of AOM should focus on the risks of rapid deposition, especially the impact of turbidity current on sediments. Full article

Dissimilatory metal-reducing bacteria are ubiquitous in soils worldwide, possess the ability to transfer electrons outside of their cell membranes, and are capable of respiring with various metal oxides. Reduction of iron oxides is one of the more energetically favorable forms of anaerobic respiration, with a higher energy yield than both sulfate reduction and methanogenesis. As such, this process has significant implications for soil carbon balances, especially in the saturated, carbon-rich soils of the northern latitudes. However, the dynamics of these microbial processes within the context of the greater soil microbiome remain largely unstudied. Previously, we have demonstrated the capability of potentiostatically poised electrodes to mimic the redox potential of iron(III)- and humic acid-compounds and obtain a measure of metal-reducing respiration. Here, we extend this work by utilizing poised electrodes to provide an inexaustable electron acceptor for iron- and humic acid-reducing microbes, and by measuring the effects on both microbial community structure and greenhouse gas emissions. The application of both nonpoised and poised graphite electrodes in peat soils stimulated methane emissions by 15%–43% compared to soils without electrodes. Poised electrodes resulted in higher (13%–24%) methane emissions than the nonpoised electrodes. The stimulation of methane emissions for both nonpoised and poised electrodes correlated with the enrichment of proteobacteria, verrucomicrobia, and bacteroidetes. Here, we demonstrate a tool for precisely manipulating localized redox conditions in situ (via poised electrodes) and for connecting microbial community dynamics with larger ecosystem processes. This work provides a foundation for further studies examining the role of dissimilatory metal-reducing bacteria in global biogeochemical cycles. Full article