In situ anoxic bioremediation is an easy-to-use technology to remediate polycyclic aromatic hydrocarbon (PAH)-contaminated soil. Degradation of PAHs mediated by soil bacteria and archaea using CO₂ as the electron acceptor is an important process for eliminating PAHs under methanogenic conditions; however, knowledge of the performance and mechanisms involved is poorly unveiled. In this study, the effectiveness and efficiency of NaHCO₃ (CO₂) as an electron acceptor to stimulate the degradation of PAHs by bacteria and archaea in highly contaminated soil were investigated. The results showed that CO₂ addition (EC2000) promoted PAH degradation compared to soil without added CO₂ (EC0), with 4.18%, 9.01%–8.05%, and 6.19%–12.45% increases for 2-, 3- and 4-ring PAHs after 250 days of incubation, respectively. Soil bacterial abundances increased with increasing incubation time, especially for EC2000 (2.90 × 10⁸ g⁻¹ soil higher than EC0, p < 0.05). Different succession patterns of the soil bacterial and archaeal communities during PAH degradation were observed. According to the PCoA and ANOSIM results, the soil bacterial communities were greatly (ANOSIM: R = 0.7232, P = 0.001) impacted by electron acceptors, whereas significant differences in the archaeal communities were not observed (ANOSIM: R = 0.553, P = 0.001). Soil bacterial and archaeal co-occurrence network analyses showed that positive correlations outnumbered the negative correlations throughout the incubation period for both treatments (e.g., EC0 and EC2000), suggesting the prevalence of coexistence/cooperation within and between these two domains rather than competition. The higher complexity, connectance, edge, and node numbers in EC2000 revealed stronger linkage and a more stable co-occurrence network compared to EC0. The results of this study could improve the knowledge on the removal of PAHs and the responses of soil bacteria and archaea to CO₂ application, as well as a scientific basis for the in situ anoxic bioremediation of PAH-contaminated industrial sites.

Biochar application into paddy is an improved strategy for addressing methane (CH₄) stimulation of straw biomass incorporation. Whereas, the differentiative patterns and mechanisms on CH₄ emission of straw biomass and biochar after long years still need to be disentangled. Considering economic feasibility, a seven-year of field experiment was conducted to explore the long-term CH₄ mitigation effect of annual low-rate biochar incorporation (RSC, 2.8 t ha⁻¹), with annual rice straw incorporation (RS, 8 t ha⁻¹) and control (CK, with no biochar or rice straw amendment incorporation) as a comparation. Results showed that RSC mitigated CH₄ emission while RS stimulated CH₄ significantly (p < 0.05) and stably over 7 experimental years compared with CK. RSC mitigated 14.8–46.7% of CH₄ emission compared with CK. In comparison to RSC, RS increased 111–950.5% of CH₄ emission during 7 field experimental years. On the 7th field experimental year, pH was significantly increased both in RS and RSC treatment (p < 0.05). RSC significantly (p < 0.05) increased soil nitrate (NO₃⁻-N) compared with RS while RS significantly (p < 0.05) increased dissolved carbon (DOC) compared to RSC. Soil NO₃⁻-N inhibition on methanogens and promotion on methanotrophs activities were verified by laboratory experiment, while soil pH and DOC mainly promoted methanogens abundance. Significantly (p < 0.05) increased DOC and soil pH enhanced methanogens growth and stimulated CH₄ emission in RS treatment. Higher soil NO₃⁻-N content in RSC than CK and RS contributed to CH₄ mitigation. Soil NO₃⁻-N and DOC were identified as the key factors differentiating CH₄ emission patterns of RS and RSC in 2019. Collectively, soil NO₃⁻-N impacts on CH₄ flux provide new ideas for prolonged effect of biochar amendment on CH₄ mitigation after years.

Arctic tundra soils store a globally significant amount of mercury (Hg), which could be transformed to the neurotoxic methylmercury (MeHg) upon warming and thus poses serious threats to the Arctic ecosystem. However, our knowledge of the biogeochemical drivers of MeHg production is limited in these soils. Using substrate addition (acetate and sulfate) and selective microbial inhibition approaches, we investigated the geochemical drivers and dominant microbial methylators in 60-day microcosm incubations with two tundra soils: a circumneutral fen soil and an acidic bog soil, collected near Nome, Alaska, United States. Results showed that increasing acetate concentration had negligible influences on MeHg production in both soils. However, inhibition of sulfate-reducing bacteria (SRB) completely stalled MeHg production in the fen soil in the first 15 days, whereas addition of sulfate in the low-sulfate bog soil increased MeHg production by 5-fold, suggesting prominent roles of SRB in Hg(II) methylation. Without the addition of sulfate in the bog soil or when sulfate was depleted in the fen soil (after 15 days), both SRB and methanogens contributed to MeHg production. Analysis of microbial community composition confirmed the presence of several phyla known to harbor microorganisms associated with Hg(II) methylation in the soils. The observations suggest that SRB and methanogens were mainly responsible for Hg(II) methylation in these tundra soils, although their relative contributions depended on the availability of sulfate and possibly syntrophic metabolisms between SRB and methanogens.

It is a common practice to maintain soil fertility based on the paddy-upland rotation with green manure in the subtropical region of China. However, rare studies are known about greenhouse gas (GHG) emissions from the paddy-upland rotation with green manure incorporation. Therefore, we conducted a field experiment of two years to compared with the effect of two kinds of green manure (CV: Chinese milk vetch and OR: Oilseed rape), and two kinds of cropping system (DR: double rice system and PR: paddy-upland rotation) on greenhouse gases emissions. We have found that the annual accumulation of CH₄ of Chinese milk vetch-rice-sweet potato || soybean was significantly reduced by 32.95%∼63.22% compared with other treatments, mainly because Chinese milk vetch reduced the abundance of methanogens by reducing soil C/N ratio. Meanwhile increasing soil permeability resulting from paddy-upland rotation also reduced soil CH₄ emission. However, The annual accumulation of N₂O of Chinese milk vetch-rice-sweet potato || soybean was increased by 17.39%∼870.11% compared with other treatments, mainly attributed to paddy-upland rotation decreased soil pH and nosZ abundance and increased nirK and nirS, thus enhancing N₂O emission, meanwhile the Chinese milk vetch incorporation and its interaction with the paddy-upland rotation has greatly enhanced the contents of NO₃⁻-N and abundance of ammonia-oxidizing archaea (AOA). The area-scaled global warming potential (GWP) and the biomass-scaled greenhouse gas emissions intensity (GHGI) of Chinese milk vetch-rice-sweet potato || soybean was reduced by 19.01%∼50.69% and 5.38%∼35.77% respectively. Thereby, the Chinese milk vetch-rice-sweet potato || soybean cropping system was suitable for agricultural sustainable development.

Methane emitted by insects is considered to be an important source of atmospheric methane. Here we report the stimulation of methane emission in the cockroach Periplaneta americana and termite Coptotermes chaohuensis, insects with abundant methanogens, by neonicotinoids, insecticides widely used to control insect pests. Cycloxaprid (CYC) and imidacloprid (IMI) caused foregut expansion in P. americana, and increased the methane emission. Antibiotics mostly eliminated the effects. In P. americana guts, hydrogen levels increased and pH values decreased, which could be significantly explained by the gut bacterium community change. The proportion of several bacterium genera increased in guts following CYC treatment, and two genera from four could generate hydrogen. Hydrogen is a central intermediate in methanogenesis. All increased methanogens in both foregut and hindgut used hydrogen as electron donor to produce methane. Besides, the up-regulation of mcrA, encoding the enzyme for the final step of methanogenesis suggested the enhanced methane production ability in present methanogens. In the termite, hydrogen levels in gut and methane emission also significantly increased after neonicotinoid treatment, which was similar to the results in P. americana. In summary, neonicotinoids changed bacterium community in P. americana gut to generate more hydrogen, which then stimulated gut methanogens to produce and emit more methane. The finding raised a new concern over neonicotinoid applications, and might be a potential environmental risk associated with atmospheric methane.

Hydrochar (HC), an environment-friendly material, enhances soil carbon sequestration and mitigate greenhouse gases (GHGs) emissions in croplands. In this study, the water-washed HC (WW-HC) was applied to paddy soil to investigate effects on nitrous oxide (N₂O) and methane (CH₄) emissions during rice growth period. Four treatments, namely control (without N fertilizer and WW-HC), N fertilizer (WW-HC00), N fertilizer with 0.5 wt% WW-HC (WW-HC05) and N fertilizer with 1.5 wt% WW-HC (WW-HC15), were established. Results showed the WW-HC addition reduced N₂O and CH₄ emissions, global warming potential (GWP) and greenhouse gas intensity (GHGI) during the growing season. Moreover, the WW-HC application reduced N₂O cumulative emission (P < 0.05) (by 28.6% and 23.8% for WW-HC05 and WW-HC15, respectively). It was mainly due to the reduced ratio of (nirK + nirS) to nosZ under WW-HC15 (P < 0.05). Compared with WW-HC00, the WW-HC05 reduced CH₄ cumulative emissions by 14.8%, while the WW-HC15 increased by 9.7%. This might be ascribed to the significantly reduced expression of the methanogenic mcrA gene and ratio of mcrA to pmoA by WW-HC (P < 0.05). The WW-HC05 amendment decreased GWP and GHGI by 18.6% and 32.5%, respectively. Furthermore, the WW-HC application greatly improved nitrogen use efficiency by 116–145% compared with the control. Our study indicates the WW-HC application is a promising GHGs mitigation practice in paddy fields.

Bacteria and archaea (prokaryotes) are vital components for maintaining healthy function of groundwater ecosystems. The prokaryotic community composition and associated putative functional processes were examined in a shallow sandy aquifer in a wet-dry tropical environment. The aquifer had a contaminated gradient of saline mine-water, which primarily consisted of elevated magnesium (Mg²⁺) and sulfate (SO₄²⁻), although other major ions and trace metals were also present. Groundwaters were sampled from piezometers, approximately 2 m in depth, located in the creek channel upstream and downstream of the mine-water influence. Sampling occurred during the dry-season when only subsurface water flow was present. Next generation sequencing was used to analyse the prokaryote assemblages using 16S rDNA and metabolic functions were predicted with FAPROTAX. Significant changes in community composition and functional processes were observed with exposure to mine-waters. Communities in the exposed sites had significantly lower relative abundance of methanotrophs such as Methylococcaceae and methanogens (Methanobacteriaceae), but higher abundance in Nitrososphaeraceae, associated with nitrification, indicating potentially important changes in the biogeochemistry of the exposed sites. The changes were most strongly correlated with concentrations of SO₄²⁻, Mg²⁺ and Na⁺. This knowledge allows an assessment of the risk of mine-water contamination to groundwater ecosystem function and aids mine-water management.

The anaerobic digestion of wastewater rich in volatile fatty acids (VFAs) provides a sustainable approach for methane production whilst reducing environmental pollution. However, the anaerobic digestion of VFAs may not be stable during long-term operation under a short hydraulic retention time. In this study, conductive carbon cloth was supplemented to investigate the impacts on the anaerobic digestion of VFAs in wastewater sourced from dark fermentation. The results demonstrated that the failure of anaerobic digestion could be avoided when carbon cloth was supplemented. In the stable stage, the methane production rate with carbon cloth supplementation was improved by 200–260%, and the chemical oxygen demand (COD) removal efficiency was significantly enhanced compared with that in the control without carbon cloth. The relative abundance of potential exoelectrogens on the carbon cloth was increased by up to 8-fold compared with that in the suspension. Electrotrophic methanogens on the carbon cloth were enriched by 4.2–17.2% compared with those in the suspension. The genera Ercella and Petrimonas along with the methanogenic archaea Methanosaeta and Methanosarcina on the carbon cloth may facilitate direct interspecies electron transfer, thereby enhancing methane production.

In this study, the YH consortium, an ethene-producing culture, was used to evaluate the effect of vitamin B₁₂ (VB₁₂) on trichloroethene (TCE) dechlorination by transferring the original TCE-reducing culture with or without adding exogenous VB₁₂. Ultra-high performance liquid chromatography - tandem mass spectrometry (UPLC-MS/MS) was applied to detect the concentrations of VB₁₂ and its lower ligand 5,6-dimethylbenzimidazole (DMB) in the cultures. After three successive VB₁₂ starvation cycles, the dechlorination of TCE stopped mostly at cis-dichloroethene (cDCE), and no ethene was found; methane production increased significantly, and no VB₁₂ was detected. Results suggest that the co-cultured microbes may not be able to provide enough VB₁₂ as a cofactor for the growth of Dehalococcoides in the YH culture, possibly due to the competition for corrinoids between Dehalococcoides and methanogens. The relative abundances of 16 S rRNA gene of Dehalococcoides and reductive dehalogenase genes tceA or vcrA were lower in the cultures without VB₁₂ compared with the cultures with VB₁₂. VB₁₂ limitation changed the microbial community structures of the consortia. In the absence of VB₁₂, the microbial community shifted from dominance of Chloroflexi to Proteobacteria after three consecutive VB₁₂ starvation cycles, and the dechlorinating genus Dehalococcoides declined from 42.9% to 13.5%. In addition, Geobacter, Clostridium, and Desulfovibrio were also present in the cultures without VB₁₂. Furthermore, the abundance of archaea increased under VB₁₂ limited conditions. Methanobacterium and Methanosarcina were the predominant archaea in the culture without VB₁₂.

Freshwater lakes, especially eutrophic ones, have become a hotspot of methanogenesis. However, the effects of eutrophication and seasonality on methanogenesis activity and methanogenic microbial community remain unclear. In the current study, for two adjacent lakes at different trophic status, their methanogenesis potential in different seasons was evaluated using incubation experiments. The density, diversity, and community structure of methanogens were analyzed based on the mcrA gene. Correlation analysis and redundancy analysis were carried out to identify the environmental factors driving the variations of methanogenesis potential and methanogen community. The results showed that eutrophication could result in active methanogenesis with relatively high seasonal variance. The methanogenesis variation could be well explained by carbon input in association with algal growth, as well as the change of methanogen population density. With the dominance of Methanomicrobiales in both lakes, the hydrogenotrophic pathway had a major contribution to total methane production. The considerable proportion of Methanomassiliicocales in eutrophic lake implied that methylotrophic methanogenesis might be previously underestimated. These results added new insights towards methanogenesis process in eutrophic freshwater lakes.