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The public and environmental health consequences of mercury (Hg) methylation have drawn much attention and considerable research to Hg methylation processes and their dynamics in diverse environmen...

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.

National audience; Failure to understand the microbial ecology driving the proliferation of antibiotic resistance in the environment prevents us from developing strategies to limit the spread of antibiotic resistant infectious disease. In this study, we developed for the first time a tyramide signal amplification-fluorescence in situ hybridization-fluorescence-activated cell sorting protocol (TSA-FISH-FACS) for the characterization of all vanA carrying bacteria in wastewater samples. Firstly, we validated the TSA-FISH protocol through microscopy in pure cultures and wastewater influent. Then, samples were sorted and quantified by FACS and qPCR. Significantly higher percentage tagging of cells was detected in vanA carrying pure cultures and wastewater samples spiked with vanA carrying cells as compared to vanA negative Gram positive strains and non-spiked wastewater samples respectively. qPCR analysis targeting vanZ, a regulating gene in the vanA cluster, showed its relative abundance was significantly greater in Enterococcus faecium ATCC 700221-spiked and positively sorted samples compared to the E. faecium spiked and negatively sorted samples. Phylogenetic analysis was then performed. Although further efforts are needed to overcome technical problems, we have, for the first time, demonstrated sorting bacterial-cells carrying antibiotic resistance genes from wastewater samples through a TSA-FISH-FACS protocol and provided insight into the microbial ecology of vancomycin resistant bacteria. Future potential applications using this approach will include the separation of members of an environmental microbial community (cultured and hard-to-culture) to allow for metagenomics on single cells or, in the case of clumping, targeting a smaller portion of the community with a priori knowledge that the target gene is present.

In groundwater, chemical reactions of Hg(II) with mineral surfaces play an important role in determining the concentration of mercury that is mobile and bioavailable. In this study, we investigated Hg(II) reduction by the ferrous carbonate mineral, siderite (FeCO 3 ), to better understand reductive transformation of mercury in anoxic carbonate-bearing waters. Kinetic experiments and X-ray adsorption spectroscopy (XAS) were conducted to examine the rate and mechanism of Hg(II) reaction with siderite. Hg(II) was reacted with synthesized siderite mineral at various concentrations and the subsequently formed Hg(0) was measured to assess the extent of mercury reduction by siderite. Our experimental data showed that Hg(II) reduction by siderite resulted in the loss of Hg when reacted with siderite mineral suspensions concurrent to formation of gaseous Hg(0). Hg(II) reduction occurred within minutes and reaction rates increased with increasing siderite surface area. XAS analysis confirmed that Hg(II) was reduced to Hg(0) and revealed that reduced mercury was sorbed to siderite surfaces suggesting that electron transfer reactions occur at siderite/water interface. The results of our study suggest that Hg(II) reduction by siderite is a kinetically favorable pathway for the mercury mobilization in ferruginous carbonate-bearing waters.

Surface water and biota from Great Salt Lake (GSL) contain some of the highest documented concentrations of total mercury (THg) and methylmercury (MeHg) in the United States. In order to identify potential biological sources of MeHg and controls on its production in this ecosystem, THg and MeHg concentrations, rates of Hg(II)-methylation and MeHg degradation, and abundances and compositions of archaeal and bacterial 16 rRNA gene transcripts were determined in sediment along a salinity gradient in GSL. Rates of Hg(II)-methylation were inversely correlated with salinity and were at or below the limits of detection in sediment sampled from areas with hypersaline surface water. The highest rates of Hg(II)-methylation were measured in sediment with low porewater salinity, suggesting that benthic microbial communities inhabiting less saline environments are supplying the majority of MeHg in the GSL ecosystem. The abundance of 16S rRNA gene transcripts affiliated with the sulfate reducer Desulfobacterium sp. was positively correlated with MeHg concentrations and Hg(II)-methylation rates in sediment, indicating a potential role for this taxon in Hg(II)-methylation in low salinity areas of GSL. Reactive inorganic Hg(II) (a proxy used for Hg(II) available for methylation) and MeHg concentrations were inversely correlated with salinity. Thus, constraints imposed by salinity on Hg(II)-methylating populations and the availability of Hg(II) for methylation are inferred to result in higher MeHg production potentials in lower salinity environments. Benthic microbial MeHg degradation was also most active in lower salinity environments. Collectively, these results suggest an important role for sediment anoxia and microbial sulfate reducers in the production of MeHg in low salinity GSL sub-habitats and may indicate a role for salinity in constraining Hg(II)-methylation and MeHg degradation activities by influencing the availability of Hg(II) for methylation.

The biological production of monomethylmercury (MeHg) in soils and sediments is an important factor controlling mercury (Hg) accumulation in aquatic and terrestrial food webs. In this study we examined the fractionation of Hg stable isotopes during Hg methylation in nongrowing cultures of the anaerobic bacteria Geobacter sulfurreducens PCA and Desulfovibrio desulfuricans ND132. Both organisms showed mass-dependent, but no mass-independent fractionation of Hg stable isotopes during Hg methylation. Despite differences in methylation rates, the two bacteria had similar Hg fractionation factors (αr/p = 1.0009 and 1.0011, respectively). Unexpectedly, δ(202)Hg values of MeHg for both organisms were 0.4‰ higher than the value of initial inorganic Hg after about 35% of inorganic Hg had been methylated. These results indicate that a (202)Hg-enriched pool of inorganic Hg was preferentially utilized as a substrate for methylation by these organisms, but that multiple intra- and/or extracellular pools supplied inorganic Hg for biological methylation. Understanding the controls of the Hg stable isotopic composition of microbially produced MeHg is important to identifying bioavailable Hg in natural systems and the interpretation of Hg stable isotopes in aquatic food webs.

Microbial methylation and demethylation are two competing processes controlling the net production and bioaccumulation of neurotoxic methylmercury (MeHg) in natural ecosystems. Although mercury (Hg) methylation by anaerobic microorganisms and demethylation by aerobic Hg-resistant bacteria have both been extensively studied, little attention has been given to MeHg degradation by anaerobic bacteria, particularly the iron-reducing bacterium Geobacter bemidjiensis Bem. Here we report, for the first time, that the strain G. bemidjiensis Bem can mediate a suite of Hg transformations, including Hg(II) reduction, Hg(0) oxidation, MeHg production and degradation under anoxic conditions. Results suggest that G. bemidjiensis utilizes a reductive demethylation pathway to degrade MeHg, with elemental Hg(0) as the major reaction product, possibly due to the presence of genes encoding homologues of an organomercurial lyase (MerB) and a mercuric reductase (MerA). In addition, the cells can strongly sorb Hg(II) and MeHg, reduce or oxidize Hg, resulting in both time and concentration-dependent Hg species transformations. Moderate concentrations (10-500 μM) of Hg-binding ligands such as cysteine enhance Hg(II) methylation but inhibit MeHg degradation. These findings indicate a cycle of Hg methylation and demethylation among anaerobic bacteria, thereby influencing net MeHg production in anoxic water and sediments.

Exposure to dietary sources of methylmercury (MeHg) is the focus of public health concerns with environmental mercury (Hg) contamination. MeHg is formed in anoxic environments by anaerobic microorganisms. This process has been studied mostly with single-species culture incubations, although the relevance of such studies to Hg(II)-methylation in situ is limited because microbial activities in the environment are critically modulated by interactions among microbial functional groups. Here we describe experiments in which Hg(II)-methylation was examined within the context of various microbial syntrophies. We show enhanced Hg(II)-methylation under conditions that established syntrophy by interspecies hydrogen and acetate transfer. Relative to activity of monocultures, interactions of Hg(II) methylating sulfate-reducing bacteria with a methanogen stimulated potential Hg(II)-methylation rates 2-fold to 9-fold, and with Syntrophobacter sp. 1.7-fold to 1.8-fold; those of a Hg(II) methylating Syntrophobacter sp. with a methanogen increased Hg(II)-methylation 2-fold. Under sulfate-depleted conditions, higher Hg(II)-methylation rates in the syntrophic incubations corresponded to higher free energy yields (ΔG°') than in the monocultures. Based on energetic considerations, we therefore propose that syntrophic microbial interactions are likely a major source of MeHg in sulfate- and iron-limited anoxic environments while in sulfate-replete environments, MeHg formation via sulfate reduction dominates.

Mercury resistant bacteria play a critical role in mercury biogeochemical cycling in that they convert methylmercury (MeHg) and inorganic mercury to elemental mercury, Hg(0). To date there are very few studies on the effects of speciation and bioavailability of MeHg in these organisms, and even fewer studies on the role that binding to cellular ligands plays on MeHg uptake. The objective of this study was to investigate the effects of thiol complexation on the uptake of MeHg by measuring the intracellular demethylation-reduction (transformation) of MeHg to Hg(0) in Hg-resistant bacteria. Short-term intracellular transformation of MeHg was quantified by monitoring the loss of volatile Hg(0) generated during incubations of bacteria containing the complete mer operon (including genes from putative mercury transporters) exposed to MeHg in minimal media compared to negative controls with non-mer or heat-killed cells. The results indicate that the complexes MeHgOH, MeHg-cysteine, and MeHg-glutathione are all bioavailable in these bacteria, and without the mer operon there is very little biological degradation of MeHg. In both Pseudomonas stutzeri and Escherichia coli, there was a pool of MeHg that was not transformed to elemental Hg(0), which was likely rendered unavailable to Mer enzymes by non-specific binding to cellular ligands. Since the rates of MeHg accumulation and transformation varied more between the two species of bacteria examined than among MeHg complexes, microbial bioavailability, and therefore microbial demethylation, of MeHg in aquatic systems likely depends more on the species of microorganism than on the types and relative concentrations of thiols or other MeHg ligands present.

Estuarine sediments support the production of monomethylmercury (MeHg) which accumulates in aquatic organisms. While natural variation in mercury stable isotope ratios can potentially be used to track sources and transformations of mercury in the environment, the isotopic signature of MeHg in sediments has not been measured directly. The isotopic composition of MeHg has been studied in laboratory experiments and fish using tandem gas chromatography-multicollector inductively coupled plasma mass spectrometry (MC–ICP-MS) systems; however, the precision and sensitivity of this method may be too low for the analysis of many environmental samples including sediments in which MeHg constitutes 1% or less of the total mercury. In this study, we developed an offline separation method for the precise measurement of the Hg isotopic composition of MeHg in estuarine sediments. Separation of MeHg from inorganic species was accomplished by distillation and chemical ethylation-GC, and was followed by gold amalgam trapping to collect and preconcentrate pyrolyzed MeHg, which was then released into an oxidizing solution. MeHg standards processed in this way were collected with an average yield of 97.5%. External precision for all replicate isotope analyses of MeHg process standards was ± 0.14‰ (2 SD, n = 8) for δ202Hg and no detectable fractionation of Hg stable isotopes occurred during the separation. δ202Hg values for MeHg separated from estuarine sediments using our approach varied from − 0.41 to + 0.41‰ and were generally higher, and spatially and temporally more variable, than those for total Hg (− 0.21 to − 0.48‰).

Ionic and organic forms of mercury (Hg) are powerful cytotoxic and neurotoxic agents in both humans and wild life. The aim of this study was to analyze the resistance profile and potential detoxification of inorganic and organic forms of Hg of bacteria isolated from marine sponges on the coast of Rio de Janeiro, Brazil. Out of the 1,236 colony forming units associated with eleven species of marine sponges, 100 morphologically different bacterial strains were analyzed in this study. Of these, 21 strains were resistant to Hg, 14 of which were classified as highly resistant because they grew despite exposure to 100 µM HgCl2. Fifteen resistant strains reduced Hg and presented merA in their genomes. The remaining six strains produced biosurfactants, suggesting that they may tolerate Hg by sequestration. Eleven strains grew in the presence of methylmercury. Our results suggest a potential for mercury detoxification by marine sponge-associated resistant bacteria, either through reduction or sequestration, as well as the possibility of bioremediation of toxic waste containing mercury.

Sponges are sessile marine invertebrates that can live for many years in the same location, and therefore, they have the capability to accumulate anthropogenic pollutants such as metals over a long period. Almost all marine sponges harbor a large number of microorganisms within their tissues. The Bacillus cereus strain Pj1 was isolated from a marine sponge, Polymastia janeirensis, and was found to be resistant to 100 μM HgCl(2) and to 10 μM methylmercury (MeHg). Pj1 was also highly resistant to other metals, including CdCl(2) and Pb(NO(3))(2), alone or in combination. The mer operon was located on the bacterial chromosome, and the volatilization test indicated that the B. cereus Pj1 was able to reduce Hg(2+)-Hg(0). Cold vapor atomic absorption spectrometry demonstrated that Pj1 volatilized 80 % of the total MeHg that it was exposed to and produced elemental Hg when incubated with 1.5 μM MeHg. Pj1 also demonstrated sensitivity to all antibiotics tested. In addition, Pj1 demonstrated a potential for biosurfactant production, presenting an emulsification activity better than synthetic surfactants. The results of this study indicate that B. cereus Pj1 is a strain that can potentially be applied in the bioremediation of HgCl(2) and MeHg contamination in aquatic environments.

Redox cycling between elemental [Hg(0)] and divalent [Hg(II)] mercury is a key control on the fate and transport of Hg in groundwater systems. In this study, we tested the ability of anaerobic bacteria to oxidize dissolved Hg(0) to Hg(II). Controlled laboratory experiments were carried out with the obligate anaerobic bacterium Geothrix fermentans H5, and the facultative anaerobic bacteria Shewanella oneidensis MR-1 and Cupriavidus metallidurans AE104. Under anoxic conditions, all three bacterial strains reacted with dissolved gaseous Hg(0) to form non-purgeable Hg. In mass balance experiments, the formation of non-purgeable Hg corresponded to the loss of volatile Hg. To determine if the non-purgeable Hg was oxidized, we performed ethylation experiments on Hg(0)-reacted cell suspensions and X-ray absorption near edge structure (XANES) spectroscopy on Hg(0)-reacted cells. Derivatization of non-purgeable Hg to diethylmercury and the Hg L III -edge position of the XANES spectra demonstrated that the reacted bacterial samples contained Hg(II). XANES analysis also revealed that cell-associated Hg(II) was covalently bound to bacterial functional groups, most likely to thiol moieties. Finally, experiments with metabolically active and heat-inactivated cells indicated that both live and dead cells oxidized Hg(0) to Hg(II). Hg(0) oxidation rates for metabolically active cultures increase in the order S. oneidensis MR-1 (1.6 × 10 − 4 fg/cell/min), C. metallidurans AE104 (2.5 × 10 − 4 fg/cell/min), and G. fermentans H5 (23.1 × 10 − 4 fg/cell/min). The results of this study suggest that reactivity towards Hg(0) is widespread among diverse anaerobic bacteria, and passive microbial oxidation of Hg(0) may play an important role in the redox transformation of mercury contaminants in subsurface environments.

In this study, we cultivated from subsurface sediments an anaerobic clostridial consortium that was composed of a fermentative Fe-reducer Clostridium species (designated as strain FGH) and a novel sulfate-reducing bacterium belonging to the clostridia family Vellionellaceae (designated as strain RU4). In pure culture, Clostridium sp. strain FGH mediated the reductive dissolution/transformation of iron oxides during growth on peptone. When Clostridium sp. FGH was grown with strain RU4 on peptone, the rates of iron oxide reduction were significantly higher. Iron reduction by the consortium was mediated by multiple mechanisms, including biotic reduction by Clostridium sp. FGH and biotic/abiotic reactions involving biogenic sulfide formed by strain RU4. The Clostridium sp. FGH produced hydrogen during fermentation, and the presence of hydrogen inhibited growth and iron reduction activity. The sulfate-reducing partner strain RU4 was stimulated by the presence of H2and generated reactive sulfide which promoted ...

The transformation of inorganic mercury (Hg) to methylmercury (MeHg) plays a key role in determining the amount of Hg that is bioaccumulated in aquatic food chains. An accurate knowledge of Hg methylation mechanisms is required to predict the conditions that promote MeHg production in aquatic environments. In this study, we conducted experiments to examine the oxidation and methylation of dissolved elemental mercury [Hg(0)] by the anaerobic bacterium Desulfovibrio desulfuricans ND132. Anoxic cultures of D. desulfuricans ND132 were exposed to Hg(0) in the dark, and samples were collected and analyzed for the loss of Hg(0), formation of non-purgeable Hg, and formation of MeHg over time. We found that D. desulfuricans ND132 rapidly transformed dissolved gaseous mercury into non-purgeable Hg, with bacterial cultures producing approximately 40 μg/L of non-purgeable Hg within 30 min, and as much as 800 μg/L of non-purgeable Hg after 36 h. Derivatization of the non-purgeable Hg in the cell suspensions to diethylmercury and analysis of Hg(0)-reacted D. desulfuricans ND132 cells using X-ray absorption near edge structure (XANES) spectroscopy demonstrated that cell-associated Hg was dominantly in the oxidized Hg(II) form. Spectral comparisons and linear combination fitting of the XANES spectra indicated that the oxidized Hg(II) was covalently bonded to cellular thiol functional groups. MeHg analyses revealed that D. desulfuricans ND132 produced up to 118 μg/L of methylmercury after 36 h of incubation. We found that a significant fraction of the methylated Hg was exported out of the cell and released into the culture medium. The results of this work demonstrate a previously unrecognized pathway in the mercury cycle, whereby anaerobic bacteria produce MeHg when provided with dissolved Hg(0) as their sole Hg source.

Mercury (Hg) is a trace metal with potentially serious public health consequences, especially when its neurotoxic form, monomethylmercury (MMHg), accumulates in aquatic and terrestrial food chains. Given the variation in Hg stable isotope abundances (up to 6‰ in δ 202/198 Hg and 11‰ in Δ 201/198 Hg) in natural environmental samples and evidence for Hg isotopic fractionation during microbial (e.g., reduction of Hg(II), degradation and formation of MMHg) and abiotic processes (e.g., photo-degradation of MMHg and volatilization, evaporation and diffusion of Hg(0)), Hg isotope biogeochemistry is currently being developed as a tool to study sources, sinks, and transformations of Hg in the environment. The use of Hg stable isotopes to identify sources of MMHg in aquatic and terrestrial food chains depends on the knowledge of reasonably precise fractionation factors for individual Hg transformations that influence speciation and accumulation of Hg. Microbial transformations are critical to the formation of MMHg and Hg(0) and to the degradation of MMHg. This article a) highlights the importance of experimental determination of microbial fractionation factors for the development of Hg stable isotope systematics because of the limitations of theoretical calculations; b) provides a summary of the current understanding of microbial stable isotope fractionation of Hg, especially during kinetically controlled processes; and c) demonstrates the various factors likely to affect cell-level fractionation during microbial transformations through use of an iterative finite-step model. We also identify future directions, conditions and controls that could help rigorously advance the development of mass-dependent Hg isotope systematics at the enzymatic, cellular and ecosystem level.

Mercury (Hg) associated with mixed waste generated by nuclear weapons manufacturing has contaminated vast areas of the Oak Ridge Reservation (ORR). Neurotoxic methylmercury (MeHg) has been formed from the inorganic Hg wastes discharged into headwaters of East Fork Poplar Creek (EFPC). Thus, understanding the processes and mechanisms that lead to Hg methylation along the flow path of EFPC is critical to predicting the impacts of the contamination and the design of remedial action at the ORR. In part I of our project, we investigated Hg(0) oxidation and methylation by anaerobic bacteria. We discovered that the anaerobic bacterium Desulfovibrio desulfuricans ND132 can oxidize elemental mercury [Hg(0)]. When provided with dissolved elemental mercury, D. desulfuricans ND132 converts Hg(0) to Hg(II) and neurotoxic methylmercury [MeHg]. We also demonstrated that diverse species of subsurface bacteria oxidizes dissolved elemental mercury under anoxic conditions. The obligate anaerobic bacterium Geothrix fermentans H5, and the facultative anaerobic bacteria Shewanella oneidensis MR-1 and Cupriavidus metallidurans AE104 can oxidize Hg(0) to Hg(II) under anaerobic conditions. In part II of our project, we established anaerobic enrichment cultures and obtained new bacterial strains from the DOE Oak Ridge site. We isolated three new bacterial strains from subsurface sediments collected from Oak Ridge.more » These isolates are Bradyrhizobium sp. strain FRC01, Clostridium sp. strain FGH, and a novel Negativicutes strain RU4. Strain RU4 is a completely new genus and species of bacteria. We also demonstrated that syntrophic interactions between fermentative bacteria and sulfate-reducing bacteria in Oak Ridge saprolite mediate iron reduction via multiple mechanisms. Finally, we tested the impact of Hg on denitrification in nitrate reducing enrichment cultures derived from subsurface sediments from the Oak Ridge site, where nitrate is a major contaminant. We showed that there is an inverse relationship between Hg concentrations and rates of denitrification in enrichment cultures. In part III of our project, we examined in more detail the effects of microbial interactions on Hg transformations. We discovered that both sulfate reducing and iron reducing bacteria coexist in freshwater sediments and both microbial groups contribute to mercury methylation. We showed that mercury methylation by sulfate reducing and iron reducing bacteria are temporally and spatially separated processes. We also discovered that methanogens can methylate mercury. We showed that Methanospirillum hungatei JF-1 methylated Hg at comparable rates, but with higher yields, than those observed for sulfate-reducing bacteria and iron-reducing bacteria. Finally, we demonstrated that syntrophic interactions between different microbial groups increase mercury methylation rates. We showed that Hg methylation rates are stimulated via inter-species hydrogen and acetate transfer (i) from sulfate-reducing bacteria to methanogens and (ii) from fermenters to the sulfate-reducing bacteria. In part IV of the project, we studied Hg bioavailability and Hg isotope fractionation. We demonstrated that thiol-bound Hg is bioavailable to mercury resistant bacteria. We found that uptake of Hg from Hg-glutathione and Hg-cysteine complexes does not require functioning glutathione and cystine/cysteine transport systems. We demonstrated that a wide range of methylmercury complexes (e.g. MeHgOH, MeHg-cysteine, and MeHg-glutathione) are bioavailable to mercury resistant bacteria. The rate of MeHg demethylation varies more between different species of mercury resistant bacteria than among MeHg complexes. We showed that microbial demethylation of MeHg depends more on the species of microorganism than on the types and relative concentrations of thiols or other MeHg ligands present. Finally, we demonstrated that Hg methylation by Geobacter sulfurreducens PCA and Desulfovibrio desulfuricans ND132 imparts mass-dependent discrimination against 202Hg relative to 198Hg. G. sulfurreducens PCA and D. desulfuricans ND132 have similar kinetic reactant/product Hg fractionation factors. Using the Hg isotope data, we showed that there are multiple intra- and/or extracellular pools provide substrate inorganic Hg for methylation.« less

Because geothermal environments contain mercury (Hg) from natural sources, microorganisms that evolved in these systems have likely adapted to this element. Knowledge of the interactions between microorganisms and Hg in geothermal systems may assist in understanding the long-term evolution of microbial adaptation to Hg with relevance to other environments where Hg is introduced from anthropogenic sources. A number of microbiological studies with supporting geochemistry have been conducted in geothermal systems across western North America. Approximately 1 in 5 study sites include measurements of Hg. Of all prokaryotic taxa reported across sites with microbiological and accompanying physicochemical data, 42% have been detected at sites in which Hg was measured. Genes specifying Hg reduction and detoxification by microorganisms were detected in a number of hot springs across the region. Archaeal-like sequences, representing two crenarchaeal orders and one order each of the Euryarchaeota and Thaumarchaeota, dominated in metagenomes' MerA (the mercuric reductase protein) inventories, while bacterial homologs were mostly found in one deeply sequenced metagenome. MerA homologs were more frequently found in metagenomes of microbial communities in acidic springs than in circumneutral or high pH geothermal systems, possibly reflecting higher bioavailability of Hg under acidic conditions. MerA homologs were found in hot springs prokaryotic isolates affiliated with Bacteria and Archaea taxa. Acidic sites with high Hg concentrations contain more of Archaea than Bacteria taxa, while the reverse appears to be the case in circumneutral and high pH sites with high Hg concentrations. However, MerA was detected in only a small fraction of the Archaea and Bacteria taxa inhabiting sites containing Hg. Nevertheless, the presence of MerA homologs and their distribution patterns in systems, in which Hg has yet to be measured, demonstrates the potential for detoxification by Hg reduction in these geothermal systems, particularly the low pH springs that are dominated by Archaea.

The contamination of groundwater with mercury (Hg) is an increasing problem worldwide. Yet, little is known about the interactions of Hg with microorganisms and their processes in subsurface environments. We tested the impact of Hg on denitrification in nitrate reducing enrichment cultures derived from subsurface sediments from the Oak Ridge Integrated Field Research Challenge site, where nitrate is a major contaminant and where bioremediation efforts are in progress. We observed an inverse relationship between Hg concentrations and onset and rates of denitrification in nitrate enrichment cultures containing between 53 and 1.1 μM of inorganic Hg; higher Hg concentrations increasingly extended the time to onset of denitrification and inhibited denitrification rates. Microbial community complexity, as indicated by terminal restriction fragment length polymorphism (tRFLP) analysis of the 16S rRNA genes, declined with increasing Hg concentrations; at the 312 nM Hg treatment, a single tRFLP peak was detected representing a culture of Bradyrhizobium sp. that possessed the merA gene indicating a potential for Hg reduction. A culture identified as Bradyrhizobium sp. strain FRC01 with an identical 16S rRNA sequence to that of the enriched peak in the tRFLP patterns, reduced Hg(II) to Hg(0) and carried merA whose amino acid sequence has 97 % identity to merA from the Proteobacteria and Firmicutes. This study demonstrates that in subsurface sediment incubations, Hg may inhibit denitrification and that inhibition may be alleviated when Hg resistant denitrifying Bradyrhizobium spp. detoxify Hg by its reduction to the volatile elemental form.

Mercury methylation and demethylation rates were measured in periphyton biofilms growing on submerged plants from a shallow fluvial lake located along the St. Lawrence River (Quebec, Canada). Incubations were performed in situ within macrophytes beds using low-level spikes of (199)HgO and Me(200)Hg stable isotopes as tracers. To determine which microbial guilds are playing a role in these processes, methylation/demethylation experiments were performed in the absence and presence of different metabolic inhibitors: chloramphenicol (general bacteriostatic inhibitor), molybdate (sodium molybdate, a sulfate reduction inhibitor), BESA (2-bromoethane sulfonic acid, a methanogenesis inhibitor), and DCMU (3-(3,4-dichlorophenyl)-1,1 dimethyl urea, a photosynthesis inhibitor). Active microbes of the periphytic consortium were also characterized using 16S rRNA gene sequencing. Methylation rates in the absence of inhibitors varied from 0.0015 to 0.0180 d(-1) while demethylation rates ranged from 0.083 to 0.217 d(-1), which corresponds to a net methylmercury balance of -0.51 to 1.28 ng gDW periphyton(-1) d(-1). Methylation rates were significantly decreased by half by DCMU and chloramphenicol, totally inhibited by BESA, and were highly stimulated by molybdate. This suggests that methanogens rather than sulfate reducing bacteria were likely the primary methylators in the periphyton of a temperate fluvial lake, a conclusion supported by the detection of 16S rRNA gene sequences that were closely related to those of methanogens. This first clear demonstration of methanogens' role in mercury methylation in environmental periphyton samples expands the known diversity of microbial guilds that contribute to the formation of the neurotoxic substance methylmercury.

It is well-established that atmospheric deposition transports mercury from lower latitudes to the Arctic. The role of bacteria in the dynamics of the deposited mercury, however, is unknown. We characterized mercury-resistant bacteria from High Arctic snow, freshwater and sea-ice brine. Bacterial densities were 9.4 × 10(5), 5 × 10(5) and 0.9-3.1 × 10(3) cells mL(-1) in freshwater, brine and snow, respectively. Highest cultivability was observed in snow (11.9%), followed by freshwater (0.3%) and brine (0.03%). In snow, the mercury-resistant bacteria accounted for up to 31% of the culturable bacteria, but

Heavy metals have been shown to negatively affect the growth of ectomycorrhizal fungi (ECMF). In addition, ECMF have been shown to accumulate heavy metals and to protect host trees from metal toxicity. However, specific literature on the interactions between ECMF and mercury (Hg) is scant. This paper describes the responses of ECMF to Hg in axenic culture conditions. Six ECMF from an area with no known history of direct Hg contamination were tested to determine their sensitivity to Hg. ECMF were incubated on solid medium amended with Hg (0–50 μM) as HgCl 2 and the effect of Hg on radial growth was determined. The effect of preexposure cultivation on Hg sensitivity, the effect of Hg on biomass production, and the ability to accumulate Hg were determined for four of the ECMF. At micromolar concentrations, Hg significantly inhibited the radial growth rate of ECMF. This inhibitory effect was lessened in some ECMF when an established colony was exposed to Hg. Mercury lowered biomass production by some ECMF, and ECMF accumulate Hg from a solid growth substrate in direct relation to the amount of Hg added to the media. Possible implications for ECMF communities in Hg-impacted areas are discussed.

Mercury (Hg) is a highly toxic element, and its contamination of groundwater presents a significant threat to terrestrial ecosystems. Understanding the geochemical processes that mediate mercury transformations in the subsurface is necessary to predict its fate and transport. In this study, we investigated the redox transformation of mercuric Hg (Hg[II]) in the presence of the Fe(II)/Fe(III) mixed valence iron oxide mineral magnetite. Kinetic and spectroscopic experiments were performed to elucidate reaction rates and mechanisms. The experimental data demonstrated that reaction of Hg(II) with magnetite resulted in the loss of Hg(II) and the formation of volatile elemental Hg (Hg[0]). Kinetic experiments showed that Hg(II) reduction occurred within minutes, with reaction rates increasing with increasing magnetite surface area (0.5 to 2 m2/L) and solution pH (4.8 to 6.7), and decreasing with increasing chloride concentration (10(-6) to 10(-2) mol/L). Mössbauer spectroscopic analysis of reacted magnetite samples revealed a decrease in Fe(II) content, corresponding to the oxidation of Fe(II) to Fe(III) in the magnetite structure. X-ray photoelectron spectroscopy detected the presence of Hg(II) on magnetite surfaces, implying that adsorption is involved in the electron transfer process. These results suggest that Hg(II) reaction with solid-phase Fe(II) is a kinetically favorable pathway for Hg(II) reduction in magnetite-hearing environmental systems.

Controlling bioaccumulation of toxic monomethylmercury (MMHg) in aquatic food chains requires differentiation between biotic and abiotic pathways that lead to its production and degradation. Recent mercury (Hg) stable isotope measurements of natural samples suggest that Hg isotope ratios can be a powerful proxy for tracing dominant Hg transforming pathways in aquatic ecosystems. Specifically, it has been shown that photo-degradation of MMHg causes both mass dependent (MDF) and mass independent fractionation (MIF) of Hg isotopes. Because the extent of MDF and MIF observed in natural samples (e.g., fish, soil and sediments) can potentially be used to determine the relative importance of pathways leading to MMHg accumulation, it is important to determine the potential role of microbial pathways in contributing to the fractionation, especially MIF, observed in these samples. This study reports the extent of fractionation of Hg stable isotopes during degradation of MMHg to volatile elemental Hg and methane via the microbial Hg resistance (mer) pathway in Escherichia coli carrying a mercury resistance (mer) genetic system on a multi-copy plasmid. During experimental microbial degradation of MMHg, MMHg remaining in reactors became progressively heavier (increasing δ202Hg) with time and underwent mass dependent Rayleigh fractionation with a fractionation factor α202/198 = 1.0004 ± 0.0002 (2SD). However, MIF was not observed in any of the microbial MMHg degradation experiments indicating that the isotopic signature left by mer mediated MMHg degradation is significantly different from fractionation observed during DOC mediated photo-degradation of MMHg. Additionally, a clear suppression of Hg isotope fractionation, both during reduction of Hg(II) and degradation of MMHg, was observed when the cell densities increased, possibly due to a reduction in substrate bioavailability. We propose a multi-step framework for understanding the extent of fractionation seen in our MMHg degradation experiments and, based on estimates of the rates of the various steps involved in this mer mediated pathway, suggest which steps in the process could contribute towards the observed extent of fractionation. This framework suggests that at lower cell densities catalysis by MerB was the rate limiting step while at higher cell densities transport into the cell, which does not cause fractionation, became the rate limiting step. In addition to presenting evidence for absence of MIF during mer mediated Hg transformations, based on the nature of Hg compounds and microbe–Hg interactions, we suggest that the nuclear spin dependent MIF (i.e., the magnetic isotope effect) is also unlikely to occur during other non mer mediated ‘dark’ microbial Hg transformations (e.g., formation of MMHg and oxidative degradation of MMHg). Because of the important implications of the absence of MIF during biological processes on Hg isotope systematics, we discuss theoretical considerations and experimental strategies that could be used to confirm this suggestion.

Mercury (Hg) stable isotope fractionation has recently been developed as a tool in biogeochemistry. In this study, the extent of Hg stable isotope fractionation during reduction of ionic mercury [Hg(II)] by two Hg(II)-resistant strains, Bacillus cereus 5 and the thermophile Anoxybacillus sp. FB9 [which actively detoxify Hg(II) by the mer system] and a Hg(II)-sensitive metal-reducing anaerobe, Shewanella oneidensis MR-1 [which reduces Hg(II) at low concentrations], was investigated. In all cases, barring suppression of fractionation that is likely due to lower Hg(II) bioavailability, the Hg(II) remaining in the reactor became progressively enriched with heavy isotopes with time and underwent mass-dependent Rayleigh fractionation with alpha202/198 values of 1.0016 +/- 0.0004 (1 SD). Based on a multistep framework for the Hg(II) reduction pathways in the three strains, we constrain the processes that could contribute toward fractionation and suggest that for Hg(II)-resistant strains, reduction by mercuric reductase is the primary step causing fractionation. The proposed framework helps explain the variation in the extent of Hg stable isotope fractionation during microbial reduction of Hg(II), furthering the promise of Hg isotope ratios as a tool in determining the role of microbial Hg transformations in the environment.

The contamination of polar regions due to the global distribution of anthropogenic pollutants is of great concern because it leads to the bioaccumulation of toxic substances, methylmercury among them, in Arctic food chains. Here we present the first evidence that microbes in the high Arctic possess and express diverse merA genes, which specify the reduction of ionic mercury [Hg(II)] to the volatile elemental form [Hg(0)]. The sampled microbial biomass, collected from microbial mats in a coastal lagoon and from the surface of marine macroalgae, was comprised of bacteria that were most closely related to psychrophiles that had previously been described in polar environments. We used a kinetic redox model, taking into consideration photoredox reactions as well as mer -mediated reduction, to assess if the potential for Hg(II) reduction by Arctic microbes can affect the toxicity and environmental mobility of mercury in the high Arctic. Results suggested that mer -mediated Hg(II) reduction could account for most of the Hg(0) that is produced in high Arctic waters. At the surface, with only 5% metabolically active cells, up to 68% of the mercury pool was resolved by the model as biogenic Hg(0). At a greater depth, because of incident light attenuation, the significance of photoredox transformations declined and merA -mediated activity could account for up to 90% of Hg(0) production. These findings highlight the importance of microbial redox transformations in the biogeochemical cycling, and thus the toxicity and mobility, of mercury in polar regions.

Mercury (Hg) undergoes systematic stable isotopic fractionation; therefore, isotopic signatures of Hg may provide a new tool to track sources, sinks, and dominant chemical transformation pathways of Hg in the environment. We investigated the isotopic fractionation of Hg by Hg(II) resistant (HgR) bacteria expressing the mercuric reductase (MerA) enzyme. The isotopic composition of both the reactant Hg(II) added to the growth medium and volatilized product (Hg(0)) was measured using cold vapor generation and multiple collector inductively coupled plasma mass spectrometry. We found that exponentially dividing pure cultures of a gram negative strain Escherichia coli JM109/pPB117 grown with abundant electron donor and high Hg(II) concentrations at 37, 30, and 22 degrees C, and a natural microbial consortium incubated in natural site water at 30 degrees C after enrichment of HgR microbes, preferentially reduced the lighter isotopes of Hg. In all cases, Hg underwent Rayleigh fractionation with the best estimates of alpha202/198 values ranging from 1.0013 to 1.0020. In the cultures grown at 37 degrees C, below a certain threshold Hg(II) concentration, the extent of fractionation decreased progressively. This study demonstrates mass-dependent kinetic fractionation of Hg and could lead to development of a new stable isotopic approach to the study of Hg biogeochemical cycling in the environment.

The contamination of polar regions with mercury that is transported as inorganic mercury from lower latitudes has resulted in the accumulation of methylmercury in the food chain of polar environments, risking the health of humans and wildlife. This problem is likely to be particularly severe in coastal marine environments where active cycling occurs. Little is currently known about how mercury is methylated in polar environments. Relating observations on mercury deposition and transport through polar regions to knowledge of the microbiology of cold environments and considering the principles of mercury transformations as have been elucidated in temperate aquatic environments, we propose that in polar regions (1) variable pathways for mercury methylation may exist, (2) mercury bioavailability to microbial transformations may be enhanced, and (3) microbial niches within sea ice are sites where active microorganisms are localized in proximity to high concentrations of mercury. Thus, microbial transformations, and consequently mercury biogeochemistry, in the Arctic and Antarctic are both unique and common to these processes in lower latitudes, and understanding their dynamics is needed for the management of mercury-contaminated polar environments.

Environmental contamination of mercury (Hg) has caused public health concerns with focuses on the neurotoxic substance methylmercury, due to its bioaccumulation and biomagnification in food chains. The goals of the present study were to examine: (i) the transformation of methylmercury, thimerosal, phenylmercuric acetate and mercuric chloride by cultures of Pseudomonas putida V1, (ii) the presence of the genes merA and merB in P. putida V1, and (iii) the degradation pathways of methylmercury by P. putida V1. Strain V1 cultures readily degraded methylmercury, thimerosal, phenylmercury acetate, and reduced mercuric chloride into gaseous Hg(0). However, the Hg transformation in LB broth by P. putida V1 was influenced by the type of Hg compounds. The merA gene was detected in P. putida V1, on the other hand, the merB gene was not detected. The sequencing of this gene, showed high similarity (100%) to the mercuric reductase gene of other Pseudomonas spp. Furthermore, tests using radioactive (14)C-methylmercury indicated an uncommon release of (14)CO2 concomitant with the production of Hg(0). The results of the present work suggest that P. putida V1 has the potential to remove methylmercury from contaminated sites. More studies are warranted to determine the mechanism of removal of methylmercury by P. putida V1.

A mer-lux bioreporter was used to study uptake of inorganic mercury, Hg(II), at trace concentrations by two facultatively anaerobic bacterial species, Vibrio anguillarumand Escherichia coli. Uptake of Hg(II) by these bacteria appeared to be facilitated, rather than by passive diffusion. Three lines of evidence support this conclusion. First, under anaerobic conditions Hg(II) uptake was greatly decreased compared with aerobic conditions, even though the chemical composition of the medium was identical except for the lack of oxygen (i.e., no reducing agents were used). Second, the uptake of Hg(II) under anaerobic conditions was not proportional to the abundance of lipophilic Hg species but was dependent on the total concentration of Hg in the samples. Third, at trace Hg(II) concentrations and under anaerobic conditions, Hg(II) uptake was enhanced by the addition of yeast extract and a variety of low molecular weight organic acids. In addition to demonstrating that Hg(II) uptake by these bacteria had the characteristics of facilitated transport, these lines of evidence also support the conclusion that processes under regulatory control of the cell affected Hg(II) uptake. If these findings apply to other bacteria as well, they mean that current models of Hg(II) uptake by microorganisms in aquatic systems, which are based solely on the role of lipophilic Hg species and passive diffusion, will need to be reconsidered.

The response of the mercury-resistant denitrifier, Pseudomonas stutzeri strain OX, and its sensitive derivative OX1 to HgCl 2 was examined under aerobic and anaerobic conditions to evaluate the pot...

Recent demonstrations of the removal and immobilization of inorganic contaminants by microbial transformations, sorption and mineralization show the potential of both natural and engineered microbes as bioremedial tools. Demonstrations of microbe-mediated mineral formation in biofilms implicate this mode of microbial life in geological evolution and remediation of inorganic contaminants.

A previously described bioassay using a mer-lux gene fusion for detection of bioavailable mercury was applied for the estimation of the bioavailable fraction of mercury in soil. The bioavailable fraction is defined here as being part of the water leachable fraction. Due to masking of light emission of soil particles leachates had to be cleaned prior to assays. Filtration of leachates through nitro-cellulose filters using pressure resulted in an underestimation of bioavailable mercury. Gravity filtration and centrifugation showed elevated (as compared with untreated leachate) and very similar responses. The utility of the mer-lux biosensor assay was tested by relating measurements of bioavailable and total mercury to the response of the soil microbial community to mercury exposure. Two different soil types (an agricultural and a beech forest soil) were spiked with 2.5 μg Hg(II) g−1 in microcosms and the frequency of mercury resistant heterotrophs and changes in community diversity, defined as the number of different 16S rDNA bands observed in DGGE gels, were monitored. In the agricultural soil the initial concentration of bioavailable mercury was estimated to be 40 ng g−1. This concentration did not change during the first 3 d and coincided with increased degrees of resistance and a decrease in diversity. The concentration of bioavailable mercury decreased subsequently rapidly and remained just above the detection level (0.2 ng g−1) for the remainder of the experiment. As a possible consequence of the decreased selection pressure of mercury, the resistance and diversity gradually returned to pre-exposure amounts. In the beech forest soil the concentration of bioavailable mercury was found to be about 20 ng g−1 throughout the experiment. This concentration did not at any time result in changes in resistance or diversity. This study showed that the bioassay using the mer-lux biosensor is a useful and sensitive tool for estimation of bioavailable mercury in soil.

Microcosms consisting of estuarine sediment slurries were examined for their utility as models for assessing effects on microbial community structure and function. Data were obtained over a 2-week period to evaluate the reproducibility between individual microcosms and the variability between microcosm slurries and fresh sediment cores. Sulfate reduction rates in microcosm slurries did not differ significantly from rates for freshly collected sediment cores (p ≥ 0.05). However, the measured rates were more variable in microcosm slurries (SE = ±0.03-0.25 nM/ml/h) than in freshly collected sediments (SE = ±0.01-0.12 nM/ml/ h). Rates of dark CO 2 fixation in the microcosm slurries declined but were consistent with rates in freshly collected sediments (6.51 and 9.29 nM/ml/h on day 3, respectively). Relative abundances (RAs) of 16S rRNA determined for six specific phylogenetic assemblages of sulfate-reducing bacteria were reproducible among three microcosm replicates with Desulfovibrio spp. consistently in greatest abundance (RA = 8.61 ± 1.40, day 7). Total direct bacterial counts were not significantly different between freshly collected sediments and microcosm slurries (p ≥ 0.05). The results indicated that microcosms were both reproducible and representative of the field, and could thus provide a potentially useful tool for studies of microbial community response to perturbation.

Elevated mercury concentrations have been reported in fish and wildlife from the Everglades in recent years. The hypothesis that eutrophication caused by the impact of phosphorous- (P) rich agricul...

Sediments from mercury-contaminated and uncontaminated reaches of the Carson River, Nevada, were assayed for sulfate reduction, methanogenesis, denitrification, and monomethylmercury (MeHg) degradation. Demethylation of [(sup14)C]MeHg was detected at all sites as indicated by the formation of (sup14)CO(inf2) and (sup14)CH(inf4). Oxidative demethylation was indicated by the formation of (sup14)CO(inf2) and was present at significant levels in all samples. Oxidized/reduced demethylation product ratios (i.e., (sup14)CO(inf2)/(sup14)CH(inf4) ratios) generally ranged from 4.0 in surface layers to as low as 0.5 at depth. Production of (sup14)CO(inf2) was most pronounced at sediment surfaces which were zones of active denitrification and sulfate reduction but was also significant within zones of methanogenesis. In a core taken from an uncontaminated site having a high proportion of oxidized, coarse-grain sediments, sulfate reduction and methanogenic activity levels were very low and (sup14)CO(inf2) accounted for 98% of the product formed from [(sup14)C]MeHg. There was no apparent relationship between the degree of mercury contamination of the sediments and the occurrence of oxidative demethylation. However, sediments from Fort Churchill, the most contaminated site, were most active in terms of demethylation potentials. Inhibition of sulfate reduction with molybdate resulted in significantly depressed oxidized/reduced demethylation product ratios, but overall demethylation rates of inhibited and uninhibited samples were comparable. Addition of sulfate to sediment slurries stimulated production of (sup14)CO(inf2) from [(sup14)C]MeHg, while 2-bromoethanesulfonic acid blocked production of (sup14)CH(inf4). These results reveal the importance of sulfate-reducing and methanogenic bacteria in oxidative demethylation of MeHg in anoxic environments.

The potential former-mediated reduction/volatilization of ionic mercury as a tool in the decontamination of a freshwater pond was evaluated using laboratory incubations and a microcosm simulation. In flask assays inoculations with ionic mercury-resistant bacteria (105−107 cells ml−1) isolated from the pond, significantly increased the rate of mercury loss (MANOVA,P≤0.05) relative to uninoculated controls. The effects of cell density, mercuric mercury concentration, addition of nutrients and supplementation with the sulfhydryl reagent β-mercaptoethanol on the rate of mercury loss, were investigated. Inoculation (by 105 cells ml−1) of a flow-through microcosm that simulated the cycling of mercury in the contaminated pond, stimulated by more than 4-fold the formation of volatile elemental mercury. Thus, biological formation of volatile mercury may hold a promise as a remedial tool of contaminated natural waters.

A microcosm consisting of water, sediment, and air compartments was used to simulate mercury geochemical cycling in a mercury-contaminated (μg L−1) pond at Oak Ridge, Tennessee. Total and dissolved mercury and total methylmercury were analyzed in water and sediment; total gaseous mercury and head-space mercury were analyzed in the water and head space, respectively. The production of gaseous mercury was correlated to dissolved mercury (0.2-μm filtration), and methylmercury was mainly produced in the sediment compartment. Addition of mercuric chloride to the system increased the production of head-space mercury by a factor of 10 but did not affect the methylation rate. Saturation of gaseous mercury in microcosm water varied from 480 to 1,500% of the solubility of elemental mercury and was controlled by unidentified factors. The microcosm maintained stable conditions for up to 3 weeks, and a mass balance indicated that it reasonably simulated the cycling of mercury in the pond. This microcosm could be used to test remedial treatments aimed at decreasing the amount of mercury that is available for accumulation by biota.

Estuarine microcosms were used to follow conjugal transfer of a broad host range IncP1 plasmid from a Pseudomonas putida donor to indigenous bacteria. Donor cells were added at a concentration similar to the natural abundance of bacteria in the water column (106 cells ml−1). Transfer was not detected in any of the test microcosms (calculated limit of detection of 10−7 and 10−4 transconjugants donor−1 in water column and sediment, respectively), with the exception of transfer to an isogenic recipient (added at 105 cells ml−1) in sediments of controls that had been inoculated with both donors and recipients. The same plasmid was transferred with high efficiencies (10−1 to 10−3) to a variety of recipients in filter and broth matings. These results suggest that if conjugal gene transfer occurred, it was at efficiencies that were not detectable in estuarine microcosms simulating natural population densities.

Environmental contextMercury, in its methylated form, is a neurotoxin that biomagnifies in marine and terrestrial foodwebs leading to elevated levels in fish and fish-eating mammals worldwide, including at numerous Arctic locations. Elevated mercury concentrations in Arctic country foods present a significant exposure risk to Arctic people. We present a detailed review of the fate of mercury in Arctic terrestrial and marine ecosystems, taking into account the extreme seasonality of Arctic ecosystems and the unique processes associated with sea ice and Arctic hydrology. AbstractThis review is the result of a series of multidisciplinary meetings organised by the Arctic Monitoring and Assessment Programme as part of their 2011 Assessment ‘Mercury in the Arctic’. This paper presents the state-of-the-art knowledge on the environmental fate of mercury following its entry into the Arctic by oceanic, atmospheric and terrestrial pathways. Our focus is on the movement, transformation and bioaccumulation of Hg in aquatic (marine and fresh water) and terrestrial ecosystems. The processes most relevant to biological Hg uptake and the potential risk associated with Hg exposure in wildlife are emphasised. We present discussions of the chemical transformations of newly deposited or transported Hg in marine, fresh water and terrestrial environments and of the movement of Hg from air, soil and water environmental compartments into food webs. Methylation, a key process controlling the fate of Hg in most ecosystems, and the role of trophic processes in controlling Hg in higher order animals are also included. Case studies on Eastern Beaufort Sea beluga (Delphinapterus leucas) and landlocked Arctic char (Salvelinus alpinus) are presented as examples of the relationship between ecosystem trophic processes and biologic Hg levels. We examine whether atmospheric mercury depletion events (AMDEs) contribute to increased Hg levels in Arctic biota and provide information on the links between organic carbon and Hg speciation, dynamics and bioavailability. Long-term sequestration of Hg into non-biological archives is also addressed. The review concludes by identifying major knowledge gaps in our understanding, including: (1) the rates of Hg entry into marine and terrestrial ecosystems and the rates of inorganic and MeHg uptake by Arctic microbial and algal communities; (2) the bioavailable fraction of AMDE-related Hg and its rate of accumulation by biota and (3) the fresh water and marine MeHg cycle in the Arctic, especially the marine MeHg cycle.

The relationship of merA gene expression (specifying the enzyme mercuric reductase) to mercury volatilization in aquatic microbial communities was investigated with samples collected at a mercury-contaminated freshwater pond, Reality Lake, in Oak Ridge, Tenn. Levels of merA mRNA transcripts and the rate of inorganic mercury [Hg(II)] volatilization were related to the concentration of mercury in the water and to heterotrophic activity in field samples and laboratory incubations of pond water in which microbial heterotrophic activity and Hg(II) concentration were manipulated. Levels of merA-specific mRNA and Hg(II) volatilization were influenced more by microbial metabolic activity than by the concentration of mercury. merA-specific transcripts were detected in some samples which did not reduce Hg(II), suggesting that rates of mercury volatilization in environmental samples may not always be proportional to merA expression.

The contamination of polar regions with mercury that is transported from lower latitudes as inorganic mercury has resulted in the accumulation of methylmercury (MeHg) in food chains, risking the health of humans and wildlife. While production of MeHg has been documented in polar marine and terrestrial environments, little is known about the responsible transformations and transport pathways and the processes that control them. We posit that as in temperate environments, microbial transformations play a key role in mercury geochemical cycling in polar regions by: (1) methylating mercury by one of four proposed pathways, some not previously described; (2) degrading MeHg by activities of mercury resistant and other bacteria; and (3) carrying out redox transformations that control the supply of the mercuric ion, the substrate of methylation reactions. Recent analyses have identified a high potential for mercury-resistant microbes that express the enzyme mercuric reductase to affect the production of gaseous elemental mercury when and where daylight is limited. The integration of microbially mediated processes in the paradigms that describe mercury geochemical cycling is therefore of high priority especially in light of concerns regarding the effect of global warming and permafrost thawing on input of MeHg to polar regions. Keywords: Microbiology; mercury biogeochemistry; redox transformations; polar regions; methylation (Published: 28 December 2011) Citation: Polar Research 2011, 30 , 15469, DOI: 10.3402/polar.v30i0.15469

We investigated microbial methylmercury (CH(3)Hg) production in sediments from the South River (SR), VA, an ecosystem contaminated with industrial mercury (Hg). Potential Hg methylation rates in samples collected at nine sites were low in late spring and significantly higher in late summer. Demethylation of (14)CH(3)Hg was dominated by (14)CH(4) production in spring, but switched to producing mostly (14)CO(2) in the summer. Fine-grained sediments originating from the erosion of river banks had the highest CH(3)Hg concentrations and were potential hot spots for both methylation and demethylation activities. Sequencing of 16S rRNA genes of cDNA recovered from sediment RNA extracts indicated that at least three groups of sulfate-reducing bacteria (SRB) and one group of iron-reducing bacteria (IRB), potential Hg methylators, were active in SR sediments. SRB were confirmed as a methylating guild by amendment experiments showing significant sulfate stimulation and molybdate inhibition of methylation in SR sediments. The addition of low levels of amorphous iron(III) oxyhydroxide significantly stimulated methylation rates, suggesting a role for IRB in CH(3)Hg synthesis. Overall, our studies suggest that coexisting SRB and IRB populations in river sediments contribute to Hg methylation, possibly by temporally and spatially separated processes.

Biosensors for the detection of pollutants in the environment can complement analytical methods by distinguishing bioavailable from inert, unavailable forms of contaminants. By using fusions of the well-understood Tn21 mercury resistance operon (mer) with promoterless luxCDABE from Vibrio fischeri, we have constructed and tested three biosensors for Hg(II). Bioluminescence specified by pRB28, carrying merRo/pT, by pOS14, mediating active transport of Hg(II), and by pOS15, containing an intact mer operon, was measured in rich and minimal media. The highest sensitivities were achieved in minimal medium and were 1, 0.5, and 25 nM Hg(II) for pRB28, pOS14, and pOS15, respectively. The utility of the biosensors in natural waters was demonstrated with freshwater, rain, and estuarine samples supplemented with Hg(II). mer-lux carried by pRB28 and pOS14 responded to Hg(II) in mercury-contaminated water samples collected from a freshwater pond. Semiquantitative analyses based on light emission in samples collected from the inlet (analytically determined total mercury, approximately 20 nM) and outlet (total mercury, approximately 7 nM) of the pond showed bioavailable mercury at approximately 20 and 1 to 2 nM, respectively. Thus, the biosensors described here semiquantitatively detect bioavailable inorganic mercury (at a nanomolar to micromolar concentration range) in contaminated waters.