The last two decades have seen nitrogen/iron-transforming bacteria at the forefront of new biogeochemical discoveries, such as anaerobic ammonium oxidation coupled to ferric iron reduction (feammox) and lithoautotrophic nitrate-reducing ferrous iron-oxidation (NRFeOx). These emerging findings continue to expand our knowledge of the nitrogen/iron cycle in nature, and also highlight the need to re-understand the functional traits of the microorganisms involved. Here, as a proof-of-principle, we report compelling evidence for the capability of a NRFeOx enrichment culture to catalyze the feammox process. Our results demonstrate that the NRFeOx culture predominantly oxidizes NH4+ to nitrogen gas, by reducing both chelated NTA-Fe(III) and poorly soluble Fe(III)-bearing minerals (γ-FeOOH) at pH 4.0 and 8.0, respectively. In the NRFeOx culture, Fe(II)-oxidizing bacteria of Rhodanobacter and Fe(III)-reducing bacteria of unclassified_Acidobacteriota coexisted. Their relative abundances were dynamically regulated by the supplemented iron sources. Metagenomic analysis revealed that the NRFeOx culture contained a complete set of denitrifying genes along with hao genes for ammonium oxidation. Additionally, numerous genes encoding extracellular electron transport-associated proteins or their homologs were identified, which facilitated the reduction of extracellular iron by this culture. More broadly, this work lightens the unexplored potential of specific microbial groups in driving nitrogen transformation through multiple pathways, and highlights the essential role of microbial iron metabolism in the integral biogeochemical nitrogen cycle.

High nitrate content limits the recycling of the secondary effluent of wastewater treatment plants. In the research, one biomass-iron mixture (BIM) filter material based on mixotrophic denitrification mode (heterotrophic and iron-driven autotrophic denitrification) was developed and used to construct a novel denitrification biological filter (BIM-DNBF) for the nitrogen removal of secondary effluent. BIM-DNBF had a short start-up time (approximately 9 days), and high total nitrogen removal (81 %-89 %) without external addition of organic carbon sources during the whole operation. The coexistence of dominant heterotrophic-denitrification-like Pseudomonas and Erysipelothrix as well as iron-driven autotrophic-denitrification-like Citrobacter, Acidovorax, etc. were found in the BIM-DNBF. Moreover, biomass was recognized as one key player in promoting the reduction of Fe3+ to Fe2+, thereby facilitating the occurrence of iron-driven autotrophic denitrification. In addition, BIM-DNBF was assessed to be affordable. These findings provide evidence that BIM-DNBF can be an efficient technology for nitrogen removal of secondary effluent.

Climate change is rapidly transforming Arctic landscapes where increasing soil temperatures speed up permafrost thaw. This exposes large carbon stocks to microbial decomposition, possibly worsening climate change by releasing more greenhouse gases. Understanding how microbes break down soil carbon, especially under the anaerobic conditions of thawing permafrost, is important to determine future changes. Here, we studied the microbial community dynamics and soil carbon decomposition potential in permafrost and active layer soils under anaerobic laboratory conditions that simulated an Arctic summer thaw. The microbial and viral compositions in the samples were analyzed based on metagenomes, metagenome-assembled genomes, and metagenomic viral contigs (mVCs). Following the thawing of permafrost, there was a notable shift in microbial community structure, with fermentative Firmicutes and Bacteroidota taking over from Actinobacteria and Proteobacteria over the 60-day incubation period. The increase in iron and sulfate-reducing microbes had a significant role in limiting methane production from thawed permafrost, underscoring the competition within microbial communities. We explored the growth strategies of microbial communities and found that slow growth was the major strategy in both the active layer and permafrost. Our findings challenge the assumption that fast-growing microbes mainly respond to environmental changes like permafrost thaw. Instead, they indicate a common strategy of slow growth among microbial communities, likely due to the thermodynamic constraints of soil substrates and electron acceptors, and the need for microbes to adjust to post-thaw conditions. The mVCs harbored a wide range of auxiliary metabolic genes that may support cell protection from ice formation in virus-infected cells.
OBJECTIVE: As the Arctic warms, thawing permafrost unlocks carbon, potentially accelerating climate change by releasing greenhouse gases. Our research delves into the underlying biogeochemical processes likely mediated by the soil microbial community in response to the wet and anaerobic conditions, akin to an Arctic summer thaw. We observed a significant shift in the microbial community post-thaw, with fermentative bacteria like Firmicutes and Bacteroidota taking over and switching to different fermentation pathways. The dominance of iron and sulfate-reducing bacteria likely constrained methane production in the thawing permafrost. Slow-growing microbes outweighed fast-growing ones, even after thaw, upending the expectation that rapid microbial responses to dominate after permafrost thaws. This research highlights the nuanced and complex interactions within Arctic soil microbial communities and underscores the challenges in predicting microbial response to environmental change.

In Southern China, the co-occurrence of arsenic (As) and antimony (Sb) contamination in soils around Sb mines presents an environmental challenge. During the flooding period of mining-impacted soils, anaerobic reduction of iron (Fe) oxides enhances the mobilization and bioavailability of Sb and As, further elevating the risk of Sb and As entering the food chain. To address this problem, activated carbon (AC) and biochar (BC) were applied to remediate flooded mining-impacted soils. Our results explored that AC can significantly decrease mobilization by 9-97 % for Sb and 9-67 % for As through inhibiting Fe(III) mineral reduction and dissolution in flooded soils. In contrast, there was no significant effect of BC. This was attributed to the strong adsorption of soil dissolved organic matter (DOM) by AC compared to BC, while DOM as electron shuttle is crucial for microbial Fe(III) reduction. Consequently, the DOM sequestration by AC effectively mitigates Sb and As leaching in contaminated mining soils.

Flooding of paddy fields during the rice growing season enhances arsenic (As) mobilization and greenhouse gas (e.g., methane) emissions. In this study, an adsorbent for dissolved organic matter (DOM), namely, activated carbon (AC), was applied to an arsenic-contaminated paddy soil. The capacity for simultaneously alleviating soil carbon emissions and As accumulation in rice grains was explored. Soil microcosm incubations and 2-year pot experimental results indicated that AC amendment significantly decreased porewater DOM, Fe(III) reduction/Fe2+ release, and As release. More importantly, soil carbon dioxide and methane emissions were mitigated in anoxic microcosm incubations. Porewater DOM of pot experiments mainly consisted of humic-like fluorophores with a molecular structure of lignins and tannins, which could mediate microbial reduction of Fe(III) (oxyhydr)oxides. Soil microcosm incubation experiments cospiking with a carbon source and AC further consolidated that DOM electron shuttling and microbial carbon source functions were crucial for soil Fe(III) reduction, thus driving paddy soil As release and carbon emission. Additionally, the application of AC alleviated rice grain dimethylarsenate accumulation over 2 years. Our results highlight the importance of microbial extracellular electron transfer in driving paddy soil anaerobic respiration and decreasing porewater DOM in simultaneously remediating As contamination and mitigating methane emission in paddy fields.

Biological nitrogen fixation (BNF) by methanotrophic bacteria has been shown to play an important role in maintaining fertility. However, this process is still limited to aerobic methane oxidation with sufficient oxygen. It has remained unknown whether and how methanotrophic BNF proceeds in hypoxic environments. Herein, we incubated paddy soils with a ferrihydrite-containing mineral salt medium to enrich methanotrophic bacteria in the presence of methane (20%, v/v) under oxygen constraints (0.27%, v/v). The resulting microcosms showed that ferrihydrite-dependent aerobic methane oxidation significantly contributed (81%) to total BNF, increasing the 15N fixation rate by 13-fold from 0.02 to 0.28 μmol 15N2 (g dry weight soil) -1 d-1. BNF was reduced by 97% when ferrihydrite was omitted, demonstrating the involvement of ferrihydrite in methanotrophic BNF. DNA stable-isotope probing indicated that Methylocystis, Methylophilaceae, and Methylomicrobium were the dominant methanotrophs/methylotrophs that assimilated labeled isotopes (13C or 15N) into biomass. Metagenomic binning combined with electrochemical analysis suggested that Methylocystis and Methylophilaceae had the potential to perform methane-induced BNF and likely utilized riboflavin and c-type cytochromes as electron carriers for ferrihydrite reduction. It was concluded that ferrihydrite mediated methanotrophic BNF by methanotrophs/methylotrophs solely or in conjunction with iron-reducing bacteria. Overall, this study revealed a previously overlooked yet pronounced coupling of iron-dependent aerobic methane oxidation to BNF and improves our understanding of methanotrophic BNF in hypoxic zones.

The biogeochemical cycling of iron (Fe) or sulfur (S) in paddy soil influences the cadmium (Cd) and arsenic (As) migration. However, the influence of coupled reduction effects and reaction precedence of Fe and S on the bioavailability of Cd and As is still not fully understood. This study aimed to reveal the influence of Fe and S reduction on soil Cd and As mobility under various pe + pH conditions and to elucidate the related mechanism in subtropical China. According to the findings, higher adsorption from Fe reduction caused high-crystalline goethite (pe + pH > 2.80) to become amorphous ferrihydrite, which in turn caused water-soluble Cd (62.0%) to first decrease. Cd was further decreased by 72.7% as a result of the transformation of SO42- to HS-/S2- via sulfate reduction and the formation of CdS and FeS. As release (an increase of 8.1 times) was consequently caused by the initial reduction and dissolution of iron oxide (pe + pH > 2.80). FeS had a lesser impact on the immobilization of As than sulfate-mediated As (V) reduction in the latter stages of the reduction process (pe + pH < 2.80). pe + pH values between 3 and 3.5 should be maintained to minimize the bioavailability of As and Cd in moderate to mildly polluted soil without adding iron oxides and sulfate amendments. The practical remediation of severely co-contaminated paddy soil can be effectively achieved by using Fe and S additions at different pe + pH conditions. This technique shows promise in reducing the bioavailability of Cd and As.

The combined contamination of cadmium (Cd) and microplastics (MPs) in paddy soil always occurred, while its influence on Cd availability remained unclear. This study investigated the Cd availability in Cd-MPs co-contaminated paddy soil in consideration of both ferric minerals and sulfate reduction under flooding conditions. The presence of MPs resulted in a higher Cd releasing risk, as represented by the increase in the available Cd and decrease in Fe-Mn oxide-bound Cd contents, especially on the 7th and 14th days based on the sequential extraction results. MPs facilitated the formation of Fe-organic ligands, which accelerated the reductive dissolution of iron minerals but decreased the amounts of amorphous iron minerals due to the release of dissolved organic substances into pore water. Furthermore, MPs promoted the relative abundance of sulfate-reducing bacteria (such as Streptomyces and Desulfovibrio genera), thus increasing the contents of reductive S species, which was advantageous to the co-precipitation of Fe, S, and Cd on the surface of MPs based on our experimental and statistical results. Taken together, both iron and sulfate reduction under anaerobic conditions played a critical role in Cd mobilization in Cd-MPs co-contaminated paddy fields.

Low-molecular-weight organic acids (LMWOAs) are essential components of rice roots exudates and an important source of soil organic carbon. The chemical-microbial pathway by which LMWOA affects arsenic (As) cycling in the rhizosphere of paddy soils is still unclear. In this study, three typical LMWOAs (acetic acid (AA), oxalic acid (OA), and citric acid (CA)) in rice root exudates were added to As-contaminated soil at a concentration of 10 mM, mimicking the rhizosphere environment. The results showed that the addition of AA and OA inhibited the mobilization of As in the rhizosphere soil. After 14 days of incubation, the content of As in the porewater of AA and OA decreased by 40% and 22%, respectively, compared with the control. AA hindered the mobilization of As in soil via promoting the formation of secondary minerals. The addition of OA inhibits the mobilization of As via increasing the proportion of As (V) in porewater and promoting the formation of secondary minerals in soil. In addition, OA addition not only significantly increased the aioA gene abundance but also notably enriched the microorganisms containing As (III) methylation functional genes (arsM). The addition of CA greatly expedited the release of As from the soil solid phase through the solubilization of Fe/Mn minerals via the effects of both soil chemistry and microbial action. Furthermore, linear discriminant analysis effect size (LEfSe) revealed the possibility that bacteria such as Burkholderia, Magnetospirillum, and Mycobacterium were involved in the reduction or methylation of As in the rhizosphere of paddy soil. This study revealed the internal causes of LMWOAs regulating As transformation and mobilization in flooded paddy soil and provided theoretical support for reducing As accumulation in rice by breeding rice varieties with high AA and OA secretions.

The iron reduction coupled with ammonia oxidation process (Feammox) is a biological reaction process associated with the nitrogen cycle that has been discovered in recent years. In this study, the iron-reducing bacterium Klebsiella sp. FC61 was attached by synthesizing nano-loadings of iron tetroxide (nFe3O4) onto rice husk biochar (RBC), and the RBC-nFe3O4 was used as an electron shuttle to participate in the biological iron reduction process of soluble and insoluble Fe3+ to improve the ammonia oxidation efficiency to 81.82%. This acceleration of electron transfer increased the carbon consumption rate and further tuned up the COD removal efficiency to 98.00%. The Feammox could be coupled with iron denitrification for internal nitrogen/iron cycling to reduce the accumulation of nitrate by-products and achieve the recycling of iron. In addition, pollutants such as Ni2+, ciprofloxacin, and formed chelates could be removed by pore adsorption and π-π interactions using bio-iron precipitates produced by iron-reducing bacteria.