Microbial Hg Methylation Research Articles

S-adenosyl-L-methionine is the unexpected methyl donor for the methylation of mercury by the membrane-associated HgcAB complex

Mercury (Hg) is a heavy metal that exhibits high biological toxicity. Monomethylmercury and dimethylmercury are neurotoxins and a significant environmental concern as they bioaccumulate and biomagnify within the aquatic food web. Microbial Hg methylation involves two proteins, HgcA and HgcB. Here, we show that HgcA and HgcB can be heterologously coexpressed, and the HgcAB complex can be purified. We demonstrated that HgcA is a membrane-associated cobalamin-dependent methyltransferase and HgcB is a ferredoxin-like protein containing two [4Fe-4S] clusters. Further, spectroscopic and kinetic results demonstrate that S-adenosyl-L-methionine (SAM) donates the methyl group to Hg in a two-step reaction involving a methylcob(III)alamin intermediate including Co-thiolate ligation from a conserved Cys residue. Our findings uncover a biological role for SAM in microbial Hg methylation.

Precipitation patterns strongly affect vertical migration and methylation of mercury in legacy contaminated sites

Legacy-contaminated sites act as significant sources of mercury (Hg) to their surrounding surface and underground environments. Intensified extreme precipitation is posing great threats to the environment and human health by changing the fate of pollutants, yet little is known about its effect on the vertical migration and methylation of Hg in contaminated sites. Here, we applied a range of simulated extreme precipitation patterns (frequency and intensity) to column leaching assays with soils collected near a contaminated site. We observed that precipitation with high frequency but low intensity resulted in more vertical migration of Hg through the soil profile than that with low frequency but high intensity. The majority (> 90%) of leached Hg was prone to migrate vertically within the top 10 cm of the soil profile. Furthermore, rainfall stimulated microbial Hg methylation, as demonstrated by enhanced production of methylmercury (MeHg) in both simulated and field-contaminated soils. We identified specific microbial taxa including Geobacteraceae, Desulfuromonadaceae, Syntrophaceae, Oscillospiraceae, and Methanomicrobiaceae as key predictors of MeHg production, which differed from those typically observed in overlying water of croplands. Particularly, the relative abundance of these dominant Hg methylators significantly increased during rainfall-induced leaching compared to that of the control, suggesting the crucial yet previously overlooked impacts of increased precipitation events on the process of microbial Hg methylation in industry-contaminated sites. Given the rising incidences of extreme precipitation events worldwide due to climate change, this study highlights the significance of assessing Hg mobility and microbial transformation in legacy contaminated sites.

Functional Genes and Transcripts Indicate the Existent and Active Microbial Mercury-Methylating Community in Mangrove Intertidal Sediments of an Urbanized Bay.

Mercury (Hg) methylation in mangrove sediments can result in the accumulation of neurotoxic methylmercury (MeHg). Identification of Hg methyltransferase gene hgcA provides the means to directly characterize the microbial Hg-methylating consortia in environments. Hitherto, the microbial Hg-methylating community in mangrove sediments was scarcely investigated. An effort to assess the diversity and abundance of hgcA genes and transcripts and link them to Hg and MeHg contents was made in the mangrove intertidal sediments along the urbanized Shenzhen Bay, China. The hgcA genes and transcripts associated with Thermodesulfobacteria [mainly Geobacteraceae, Syntrophorhabdaceae, Desulfobacterales, and Desulfarculales (these four lineages were previously classified into the Deltaproteobacteria taxon)], as well as Euryarchaeota (mainly Methanomicrobia and Theionarchaea) dominated the hgcA-harboring communities, while Chloroflexota, Nitrospirota, Planctomycetota, and Lentisphaerota-like hgcA sequences accounted for a small proportion. The hgcA genes appeared in greater abundance and diversity than their transcript counterparts in each sampling site. Correlation analysis demonstrated that the MeHg content rather than Hg content significantly correlated with the structure of the existent/active hgcA-harboring community and the abundance of hgcA genes/transcripts. These findings provide better insights into the microbial Hg methylation drivers in mangrove sediments, which could be helpful for understanding the MeHg biotransformation therein.

Insights into the reduction of methylmercury accumulation in rice grains through biochar application: Hg transformation, isotope fractionation, and transcriptomic analysis

Methylmercury (MeHg), a potent neurotoxin, easily moves from the soil into rice plants and subsequently accumulates within the grains. Although biochar can reduce MeHg accumulation in rice grains, the precise mechanism underlying biochar-mediated responses to mercury (Hg) stress, specifically regarding MeHg accumulation in rice, remains poorly understood. In the current study, we employed a 4% biochar amendment to remediate Hg-contaminated paddy soil, elucidate the impacts of biochar on MeHg accumulation through a comprehensive analysis involving Hg isotopic fractionation and transcriptomic analyses. The results demonstrated that biochar effectively lowered the levels of MeHg in paddy soils by decreasing bioavailable Hg and microbial Hg methylation. Furthermore, biochar reduced the uptake and translocation of MeHg in rice plants, ultimately leading to a reduction MeHg accumulation in rice grains. During the process of total mercury (THg) uptake, biochar induced a more pronounced negative isotope fractionation magnitude, whereas the effect was less pronounced during the upward transport of THg. Conversely, biochar caused a more pronounced positive isotope fractionation magnitude during the upward transport of MeHg. Transcriptomics analyses revealed that biochar altered the expression levels of genes associated with the metabolism of cysteine, glutathione, and metallothionein, cell wall biogenesis, and transport, which possibly enhance the sequestration of MeHg in rice roots. These findings provide novel insights into the effects of biochar application on Hg transformation and transport, highlighting its role in mitigating MeHg accumulation in rice.

Effects of vegetation on methylmercury concentrations and loads in a mercury contaminated floodplain

The Yolo Bypass (YB) is a large flood conveyance system designed to protect the city of Sacramento, California, USA from flooding when the Sacramento River approaches flood stage. The Sacramento River watershed and YB are a source of methylmercury (MeHg) to downstream habitat as a result of historic mercury (Hg) and gold mining practices. In the dry season, the YB is extensively farmed and grazed. However, depending on the water year, the floodplain may remain inundated for months. Our experiments focused on the role of pasture land and decomposing vegetation as a source of MeHg during extensive periods of floodplain flooding. Decomposing vegetation, rather than sediment, was identified as the principal source of filter passing MeHg (fMeHg) within the floodplain. The decomposing vegetation provided a substrate for microbial methylation of inorganic Hg contained within the plants. In replicated flooded mesocosm experiments, MeHg concentrations increased from 2.78 to 31.0 ng g−1 dw and 3.41 to 56.8 ng g−1 dw in decomposing vegetation. In field collections, the concentrations of MeHg in vegetation increased from preflood levels of 2.78 to 45.4 ng g−1 dw after 17 weeks of flooding. The importance of vegetation was shown in laboratory experiments where there was a positive correlation between the amount of fMeHg in water and the amount of vegetation added. These results also provide Hg concentration data for an important functional type of vegetation, grasses, and fill a data gap that contributed to uncertainties with regards to the role of vegetation in Hg cycling.

Coupling fluorescent probes to characterize S-containing compounds in a sulfate reducing bacteria involved in Hg methylation

The microbial methylation of inorganic mercury Hg(II) is governed by S-containing compounds such as thiols (RSH) and sulfides (S2−). Various S-containing molecules in an environmental or culture medium can be difficult to assess because of the complexity of the medium, poor stability, and low concentration ranges of sulfide and thiol compounds. Here, we applied two fluorescence spectroscopy-based methods using α, β-unsaturated ethanoylcoumarin fluorophore (DHC) for the quantification of sulfides, and monobromo (trimethylammonio) bimane (qBBr) to quantify total thiol concentrations (in extracellular and bacterial cell fractions). The potential interferences of both organic and inorganic compounds from the matrix were evaluated. In the presence of Hg species, both methods allowed the quantification of free sulfides or thiols (not forming complexes with Hg). The two methods were highly sensitive, with detection limits of 100 nM and 20 nM for thiols and sulfides, respectively. They also exhibited high selectivity for the detection of thiols or sulfides against other tested matrix compounds. Finally, both methods were applied to characterize S-containing compounds in a culture of Pseudodesulfovibrio hydrargyri strain BerOc1, a methylating sulfate-reducing bacterium (SRB) exposed to 0.1 mM of cysteine. During bacterial growth, we used (i) DHC probe to quantify sulfide concentration in the bulk fraction, (ii) qBBr for total extracellular thiols and total thiols adsorbed on the cells, and (iii) liquid chromatography-tandem mass spectrometry to track cysteine degradation and characterize other thiols. The time series until the end of BerOc1 growth showed biodegradation of cysteine, and biosynthesis of sulfides and other thiol compounds.

Environmental formation of methylmercury is controlled by synergy of inorganic mercury bioavailability and microbial mercury-methylation capacity.

Methylmercury (MeHg) production is controlled by the bioavailability of inorganic divalent mercury (Hg(II)i ) and Hg-methylation capacity of the microbial community (conferred by the hgcAB gene cluster). However, the relative importance of these factors and their interaction in the environment remain poorly understood. Here, metagenomic sequencing and a full-factorial MeHg formation experiment were conducted across a wetland sulfate gradient with different microbial communities and pore water chemistries. From this experiment, the relative importance of each factor on MeHg formation was isolated. Hg(II)i bioavailability correlated with the dissolved organic matter composition, while the microbial Hg-methylation capacity correlated with the abundance of hgcA genes. MeHg formation responded synergistically to both factors. Notably, hgcA sequences were from diverse taxonomic groups, none of which contained genes for dissimilatory sulfate reduction. This work expands our understanding of the geochemical and microbial constraints on MeHg formation in situ and provides an experimental framework for further mechanistic studies. This article is protected by copyright. All rights reserved.

Mercury methylation potential and bioavailability in the sediments of two distinct aquatic systems

This study explored mercury (Hg) methylation potential in two distinct aquatic systems. Fourmile Creek (FMC) was historically polluted with Hg effluents from groundwater as it is a typical gaining stream, where organic matter and microorganisms in streambed are continuously winnowed. The H02 constructed wetland only receives atmospheric Hg and is rich in organic matter and microorganisms. Both systems receive Hg from atmospheric deposition now. Surface sediments were collected from FMC and H02, spiked with inorganic Hg, and cultivated in an anaerobic chamber to stimulate microbial Hg methylation reactions. Total mercury (THg) and methylmercury (MeHg) concentrations were measured at each spiking stage. Mercury methylation potential (MMP, %MeHg in THg) and Hg bioavailability were assessed with the deployment of diffusive gradients in thin films (DGTs). During the methylation process and at the same incubation stage, FMC sediment showed faster increasing rates of %MeHg and higher MeHg concentrations than H02, demonstrating a stronger MMP in the FMC sediment. Similarly, higher Hg bioavailability was observed in FMC sediment compared to the H02 as indicated by DGT-Hg concentrations. In conclusion, the H02 wetland with high levels of organic matter and microorganisms presented low MMP. But the Fourmile Creek as a gaining stream and a historical site of Hg pollution showed strong MMP and high Hg bioavailability. A related study on microbial community activities characterized the microorganisms between FMC and H02, which is attributed to be the main reason for their different methylation capabilities. Our study further brought up the considerations on remediated sites from Hg contamination: Hg bioaccumulation and biomagnification can still be elevated and higher than the surrounding environment due to lagged changes in microbial community structures. This study supported the sustainable ecological modifications of legacy Hg contamination and raised the necessity of long-term monitoring actions even after executing a remediation plan.

"Trojan Horse" Type Internalization Increases the Bioavailability of Mercury Sulfide Nanoparticles and Methylation after Intracellular Dissolution.

Mercury sulfide nanoparticles (HgSNP), as natural metal-containing nanoparticles, are the dominant Hg species in anoxic zones. Although the microbial Hg methylation of HgSNP has been previously reported, the importance of this process in Hg methylation has yet to be clarified due to the lack of knowledge on the internalization and transformation of HgSNP. Here, we investigated the internalization and transformation of HgSNP in microbial methylator Geobacter sulfurreducens PCA through total Hg analysis and different Hg species quantification in medium and cytoplasm. We found that the microbial uptake of HgSNP, via a passive diffusion pathway, was significantly higher than that of the Hg2+-dissolved organic matter (Hg2+-DOM) complex. Internalized HgSNP were dissolved to Hg2+ in cytoplasm with a maximal dissolution of 41%, suggesting a "Trojan horse" mechanism. The intracellular Hg2+ from HgSNP exposure at the initial stage (8 h) was higher than that in Hg2+-DOM group, which led to higher methylation of HgSNP. Furthermore, no differences in methylmercury (MeHg) production from HgSNP were observed between the hgcAB gene knockout (ΔhgcAB) and wild-type strains, suggesting that HgSNP methylation may occur through HgcAB-independent pathways. Considering the possibility of a broad range of hgcAB-lacking microbes serving as methylators for HgSNP and the ubiquity of HgSNP in anoxic environments, this study highlights the importance of HgSNP internalization and methylation in MeHg production and demonstrates the necessity of understanding the assimilation and transformation of nutrient and toxic metal nanoparticles in general.

Potential for mercury methylation by Asgard archaea in mangrove sediments.

Methylmercury (MeHg) is a potent neurotoxin that bioaccumulates along food chains. The conversion of MeHg from mercury (Hg) is mediated by a variety of anaerobic microorganisms carrying hgcAB genes. Mangrove sediments are potential hotspots of microbial Hg methylation; however, the microorganisms responsible for Hg methylation are poorly understood. Here, we conducted metagenomic and metatranscriptomic analyses to investigate the diversity and distribution of putative microbial Hg-methylators in mangrove ecosystems. The highest hgcA abundance and expression occurred in surface sediments in Shenzhen, where the highest MeHg concentration was also observed. We reconstructed 157 metagenome-assembled genomes (MAGs) carrying hgcA and identified several putative novel Hg-methylators, including one Asgard archaea (Lokiarchaeota). Further analysis of MAGs revealed that Deltaproteobacteria, Euryarchaeota, Bacteroidetes, Chloroflexi, and Lokiarchaeota were the most abundant and active Hg-methylating groups, implying their crucial role in MeHg production. By screening publicly available MAGs, 104 additional Asgard MAGs carrying hgcA genes were identified from a wide range of coast, marine, permafrost, and lake sediments. Protein homology modelling predicts that Lokiarchaeota HgcAB proteins contained the highly conserved amino acid sequences and folding structures required for Hg methylation. Phylogenetic tree revealed that hgcA genes from Asgard clustered with fused hgcAB genes, indicating a transitional stage of Asgard hgcA genes. Our findings thus suggest that Asgard archaea are potential novel Hg-methylating microorganisms and play an important role in hgcA evolution.

Distribution and phylogeny of mercury methylation, demethylation, and reduction genes in the Seto Inland Sea of Japan

Mercury (Hg) adversely affects human and environmental health. To evaluate the mercury (Hg) speciation (methylation, demethylation, and reduction) of microorganisms in coastal seawater, we analyzed the microbial functional gene sets involved in Hg methylation (hgcA and hgcB), demethylation (merB), and reduction (merA) using a metagenomic approach in the eastern and western parts (the Kii and Bungo channels, respectively) of the Seto Inland Sea (SIS) of Japan. We determined the concentration of dissolved total mercury (dTHg) and methylated mercury (dMeHg) in seawater. The metagenomic analysis detected hgcAB, merA, and merB in both channels, whereas the phylogenies of these genes differed between them. A correlation between Hg concentration (both dTHg and dMeHg) and the relative abundance of each gene was not observed. Our data suggests that microbial Hg methylation and demethylation could occur in the SIS and there could be a distinct microbial Hg speciation process between the Kii and Bungo channels.

Ferrous sulfide nanoparticles control mercury speciation and bioavailability to methylating bacteria in contaminated groundwater: Impacts of mercury species

Mercury speciation in groundwater affects its removal effectiveness and methylation potential. Yet, most studies focus on the removal of inorganic dissolved Hg(II) and few studies explored the mercury methylation before and after the treatment. This study comprehensively explored the removal performance of three model mercury species, namely, dissolved inorganic divalent Hg (Hg(II), including free Hg2+ and Hg2+ complexes with Cl− and OH−), Hg2+ bound to dissolved organic matter (Hg-DOM), and HgS nanoparticles by FeS nanoparticles and further investigated the resultant impacts on the microbial methylation of Hg. Among three different stabilizers (starch, carboxymethyl cellulose (CMC), carboxymethyl starch (CMS)), CMC stabilized FeS nanoparticles (CMC-FeS) demonstrated best physical stability and highest mercury uptake. The CMC-FeS nanoparticles efficiently immobilized the three mercury species within 20 h. The sorption isotherm data of Hg(II) and Hg-DOM were well fitted by the dual-mode isotherm model and the maximum sorption capacities were 3358.28 and 2396.38 mg/g, respectively. Hg(II) and Hg-DOM were predominantly removed via ion exchange, chemical precipitation, and surface complexation whereas HgS was mainly immobilized through heteroaggregation. The simple treatment greatly reduced the bioavailable Hg species, thereby diminishing the net MeHg production by 70.2 %, 32.7 %, and 11.3 %, respectively. This study provides compelling evidence that FeS nanoparticles efficiently removed various mercury species in groundwater and remarkably inhibited the microbial methylation of mercury.

Declines of methylmercury along a salinity gradient in a low-lying coastal wetland ecosystem at South Carolina, USA

Wetlands are widely regarded as biogeochemical hotspots of highly toxic methylmercury (MeHg), mainly mediated by sulfate-reducing bacteria. In low-lying coastal wetlands, sea level rise, a phenomenon caused by global climate change, is slowly degrading numerous healthy freshwater forested wetlands into salt-degraded counterparts with a nickname “ghost forests”, and eventually converting them to saltmarshes. However, little is known about the changes of mercury (Hg) methylation, bioaccumulation, and biomagnification along the forest-to-saltmarsh gradient. Here, we conducted extensive field sampling in three wetland states (healthy forested wetlands, salt-degraded forested wetlands, and saltmarsh) along a salinity gradient (from 0 to 9.4 ppt) in Winyah Bay, South Carolina, USA. We found that in our study wetland systems the saltmarshes had the lowest levels of both total Hg and MeHg in sediments and biota, as compared to healthy forested wetlands and saltwater-degraded ghost forests. Our results suggest that the slow conversion of forested wetland to saltmarsh could reduce net MeHg production in our study wetland systems, which we hypothesized that could be attributed to increased sulfate reduction and excessive buildup of sulfide in sediment that inhibits microbial Hg methylation, and/or reduced canopy density and increased photodegradation of MeHg. However, it should be noted that biogeochemical MeHg responses to salinity changes may be site-specific and we urge more similar studies in other wetland systems along a salinity gradient. Therefore, long-term salinization of coastal wetlands and the slow conversion of forests to marshes could decrease long-term exposure of toxic MeHg levels in coastal food webs that are similar to our system, and ultimately reduce human exposure to this neurotoxin.

Expression Levels of hgcAB Genes and Mercury Availability Jointly Explain Methylmercury Formation in Stratified Brackish Waters.

Neurotoxic methylmercury (MeHg) is formed by microbial methylation of inorganic divalent Hg (HgII) and constitutes severe environmental and human health risks. The methylation is enabled by hgcA and hgcB genes, but it is not known if the associated molecular-level processes are rate-limiting or enable accurate prediction of MeHg formation in nature. In this study, we investigated the relationships between hgc genes and MeHg across redox-stratified water columns in the brackish Baltic Sea. We showed, for the first time, that hgc transcript abundance and the concentration of dissolved HgII-sulfide species were strong predictors of both the HgII methylation rate and MeHg concentration, implying their roles as principal joint drivers of MeHg formation in these systems. Additionally, we characterized the metabolic capacities of hgc+ microorganisms by reconstructing their genomes from metagenomes (i.e., hgc+ MAGs), which highlighted the versatility of putative HgII methylators in the water column of the Baltic Sea. In establishing relationships between hgc transcripts and the HgII methylation rate, we advance the fundamental understanding of mechanistic principles governing MeHg formation in nature and enable refined predictions of MeHg levels in coastal seas in response to the accelerating spread of oxygen-deficient zones.

Increased water inputs fuel microbial mercury methylation in upland soils

Mercury (Hg) can be converted to neurotoxic methylmercury (MeHg) by certain microbes typically in anaerobic environments, threatening human health due to its bioaccumulation in food webs. However, it is unclear whether and how Hg can be methylated in legacy aerobic uplands with increasing water. Here, we conducted a series of incubation experiments to investigate the effects of increased water content on MeHg production in two typical upland soils (i.e., long-term and short-term use). Results showed that marked MeHg production occurred in water-saturated upland soils, which was strongly correlated with the proportions of significantly stimulated Hg methylating taxon (i.e., Geobacter). Elevated temperature further enhanced MeHg production by blooming proportions of typical Hg methylators (i.e., Clostridium, Acetonema, and Geobacter). Water saturation could also enhance microbial Hg methylation by facilitating microbial syntrophy between non-Hg methylators and Hg methylators. Taken together, the present work suggests that uplands could turn into a potential MeHg reservoir in response to water inputs resulting from rainfall or anthropogenic irrigation.

Microbial mercury methylation potential in a large-scale municipal solid waste landfill, China

Landfills harbor ideal conditions for microbial mercury methylation. However, the levels and distribution of mercury (Hg) and methylmercury (MeHg), potential microbial Hg methylation, and their linkage within landfills are largely unknown. In the present study, total mercury (THg), MeHg, the Hg methylation gene (hgcA) and mer operon were quantified in 30 waste samples from different depths (0–30 m) at 5 locations within a large-scale landfill in China. The average concentrations of THg and MeHg in the solid waste samples were 1422.91 ng/g and 3.15 ng/g, respectively. THg (up to 14405.29 ng/g) and MeHg (up to 10.42 ng/g) have high concentrations in the middle part (10–15 m) along the depth profiles. The concentration of THg was strongly positively (both p < 0.05) correlated with the MeHg concentration and the relative abundance of hgcA, indicating that the THg concentration can play an important role in microbial Hg methylation. The hgcA genes were detected in most samples and mer operon were detected in all samples. Combined hgcA qPCR and metagenomics data showed that Archaea Methanofollis may mainly account for Hg methylation within landfills. These findings provide fundamental knowledge on Hg cycles in landfills.

Experimental evidence for recovery of mercury-contaminated fish populations

Anthropogenic releases of mercury (Hg)1–3 are a human health issue4 because the potent toxicant methylmercury (MeHg), formed primarily by microbial methylation of inorganic Hg in aquatic ecosystems, bioaccumulates to high concentrations in fish consumed by humans5,6. Predicting the efficacy of Hg pollution controls on fish MeHg concentrations is complex because many factors influence the production and bioaccumulation of MeHg7–9. Here we conducted a 15-year whole-ecosystem, single-factor experiment to determine the magnitude and timing of reductions in fish MeHg concentrations following reductions in Hg additions to a boreal lake and its watershed. During the seven-year addition phase, we applied enriched Hg isotopes to increase local Hg wet deposition rates fivefold. The Hg isotopes became increasingly incorporated into the food web as MeHg, predominantly from additions to the lake because most of those in the watershed remained there. Thereafter, isotopic additions were stopped, resulting in an approximately 100% reduction in Hg loading to the lake. The concentration of labelled MeHg quickly decreased by up to 91% in lower trophic level organisms, initiating rapid decreases of 38–76% of MeHg concentration in large-bodied fish populations in eight years. Although Hg loading from watersheds may not decline in step with lowering deposition rates, this experiment clearly demonstrates that any reduction in Hg loadings to lakes, whether from direct deposition or runoff, will have immediate benefits to fish consumers.

Oxygen‐deficient water zones in the Baltic Sea promote uncharacterized Hg methylating microorganisms in underlying sediments

AbstractHuman‐induced expansion of oxygen‐deficient zones can have dramatic impacts on marine systems and its resident biota. One example is the formation of the potent neurotoxic methylmercury (MeHg) that is mediated by microbial methylation of inorganic divalent Hg (HgII) under oxygen‐deficient conditions. A negative consequence of the expansion of oxygen‐deficient zones could be an increase in MeHg production due to shifts in microbial communities in favor of microorganisms methylating Hg. There is, however, limited knowledge about Hg‐methylating microbes, i.e., those carrying hgc genes critical for mediating the process, from marine sediments. Here, we aim to study the presence of hgc genes and transcripts in metagenomes and metatranscriptomes from four surface sediments with contrasting concentrations of oxygen and sulfide in the Baltic Sea. We show that potential Hg methylators differed among sediments depending on redox conditions. Sediments with an oxygenated surface featured hgc‐like genes and transcripts predominantly associated with uncultured Desulfobacterota (OalgD group) and Desulfobacterales (including Desulfobacula sp.) while sediments with a hypoxic‐anoxic surface included hgc‐carrying Verrucomicrobia, unclassified Desulfobacterales, Desulfatiglandales, and uncharacterized microbes. Our data suggest that the expansion of oxygen‐deficient zones in marine systems may lead to a compositional change of Hg‐methylating microbial groups in the sediments, where Hg methylators whose metabolism and biology have not yet been characterized will be promoted and expand.

Nutrient Inputs Stimulate Mercury Methylation by Syntrophs in a Subarctic Peatland

Climate change dramatically impacts Arctic and subarctic regions, inducing shifts in wetland nutrient regimes as a consequence of thawing permafrost. Altered hydrological regimes may drive changes in the dynamics of microbial mercury (Hg) methylation and bioavailability. Important knowledge gaps remain on the contribution of specific microbial groups to methylmercury (MeHg) production in wetlands of various trophic status. Here, we measured aqueous chemistry, potential methylation rates (kmeth), volatile fatty acid (VFA) dynamics in peat-soil incubations, and genetic potential for Hg methylation across a groundwater-driven nutrient gradient in an interior Alaskan fen. We tested the hypotheses that (1) nutrient inputs will result in increased methylation potentials, and (2) syntrophic interactions contribute to methylation in subarctic wetlands. We observed that concentrations of nutrients, total Hg, and MeHg, abundance of hgcA genes, and rates of methylation in peat incubations (kmeth) were highest near the groundwater input and declined downgradient. hgcA sequences near the input were closely related to those from sulfate-reducing bacteria (SRB), methanogens, and syntrophs. Hg methylation in peat incubations collected near the input source (FPF2) were impacted by the addition of sulfate and some metabolic inhibitors while those down-gradient (FPF5) were not. Sulfate amendment to FPF2 incubations had higher kmeth relative to unamended controls despite no effect on kmeth from addition of the sulfate reduction inhibitor molybdate. The addition of the methanogenic inhibitor BES (25 mM) led to the accumulation of VFAs, but unlike molybdate, it did not affect Hg methylation rates. Rather, the concurrent additions of BES and molybdate significantly decreased kmeth, suggesting a role for interactions between SRB and methanogens in Hg methylation. The reduction in kmeth with combined addition of BES and molybdate, and accumulation of VFA in peat incubations containing BES, and a high abundance of syntroph-related hgcA sequences in peat metagenomes provide evidence for MeHg production by microorganisms growing in syntrophy. Collectively the results suggest that wetland nutrient regimes influence the activity of Hg methylating microorganisms and, consequently, Hg methylation rates. Our results provide key information about microbial Hg methylation and methylating communities under nutrient conditions that are expected to become more common as permafrost soils thaw.

Algal Organic Matter Drives Methanogen-Mediated Methylmercury Production in Water from Eutrophic Shallow Lakes.

Algal blooms bring massive amounts of algal organic matter (AOM) into eutrophic lakes, which influences microbial methylmercury (MeHg) production. However, because of the complexity of AOM and its dynamic changes during algal decomposition, the relationship between AOM and microbial Hg methylators remains poorly understood, which hinders predicting MeHg production and its bioaccumulation in eutrophic shallow lakes. To address that, we explored the impacts of AOM on microbial Hg methylators and MeHg production by characterizing dissolved organic matter with Fourier transform ion cyclotron resonance mass spectrometry (FTICR-MS) and three-dimensional excitation-emission matrix (3D-EEM) fluorescence spectroscopy and quantifying the microbial Hg methylation gene hgcA. We first reveal that the predominance of methanogens, facilitated by eutrophication-induced carbon input, could drive MeHg production in lake water. Specifically, bioavailable components of AOM (i.e., CHONs such as aromatic proteins and soluble microbial byproduct-like materials) increased the abundances (Archaea-hgcA gene: 438-2240% higher) and activities (net CH4 production: 16.0-44.4% higher) of Archaea (e.g., methanogens). These in turn led to enhanced dissolved MeHg levels (24.3-15,918% higher) for three major eutrophic shallow lakes in China. Nevertheless, our model results indicate that AOM-facilitated MeHg production could be offset by AOM-induced MeHg biodilution under eutrophication. Our study would help reduce uncertainties in predicting MeHg production, providing a basis for mitigating the MeHg risk in eutrophic lakes.