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Mercury (Hg) methylation, methylmercury (MeHg) demethylation, and inorganic redox transformations of Hg are microbe-mediating processes that determine the fate and cycling of Hg and MeHg in many environments, and by doing so influence the health of humans and wild life. The discovery of the Hg methylation genes, hgcAB, in the last decade together with advances in high throughput and genome sequencing methods, have resulted in an expanded appreciation of the diversity of Hg methylating microbes. This review aims to describe experimentally confirmed and recently discovered hgcAB gene-carrying Hg methylating microbes; phylogenetic and taxonomic analyses are presented. In addition, the current knowledge on transformation mechanisms, the organisms that carry them out, and the impact of environmental parameters on Hg methylation, MeHg demethylation, and inorganic Hg reduction and oxidation is summarized. This knowledge provides a foundation for future action toward mitigating the impact of environmental Hg pollution.

This is a case-study of an organized Israeli effort to disengage the issue of corporations in the Palestinian Occupied Territories from the global CSR framework of human rights. At the centre of th...

Mercury (Hg) is a highly toxic element due to its high affinity for protein sulfhydryl groups, which upon binding, can destabilize protein structure and decrease enzyme activity. Prokaryotes have evolved enzymatic mechanisms to detoxify inorganic Hg and organic Hg (e.g., MeHg) through the activities of mercuric reductase (MerA) and organomercury lyase (MerB), respectively. Here, the taxonomic distribution and evolution of MerAB was examined in 84,032 archaeal and bacterial genomes, metagenome assembled genomes, and single-cell genomes. Homologs of MerA and MerB were identified in 7.8 and 2.1% percent of genomes, respectively. MerA was identified in the genomes of 10 archaeal and 28 bacterial phyla previously unknown to code for this functionality. Likewise, MerB was identified in 2 archaeal and 11 bacterial phyla previously unknown to encode this functionality. Surprisingly, homologs of MerB were identified in a number of genomes (∼50% of all MerB-encoding genomes) that did not encode MerA, suggesting alternative mechanisms to detoxify Hg(II) once it is generated in the cytoplasm. Phylogenetic reconstruction of MerA place its origin in thermophilic Thermoprotei (Crenarchaeota), consistent with high levels of Hg(II) in geothermal environments, the natural habitat of this archaeal class. MerB appears to have been recruited to the mer operon relatively recently and likely among a mesophilic ancestor of Euryarchaeota and Thaumarchaeota. This is consistent with the functional dependence of MerB on MerA and the widespread distribution of mesophilic microorganisms that methylate Hg(II) at lower temperature. Collectively, these results expand the taxonomic and ecological distribution of mer-encoded functionalities, and suggest that selection for Hg(II) and MeHg detoxification is dependent not only on the availability and type of mercury compounds in the environment but also the physiological potential of the microbes who inhabit these environments. The expanded diversity and environmental distribution of MerAB identify new targets to prioritize for future research.

Yellowstone National Park is home to many different hot springs, lakes, geysers, pools, and basins that range in pH, chemical composition, and temperature. These different environmental variations provide a broad range of conditions that select and grow diverse communities of microorganisms. In this study, we collected samples from geochemically diverse lakes and springs to characterize the microbial communities present through 16S rRNA metagenomic analysis. This information was then used to observe how various microorganisms survive in high mercury environments. The results show the presence of microorganisms that have been studied in previous literature. The results also depict gradients of microorganisms including thermophilic bacteria and archaea that exist in these extreme environments. In addition, beta diversity analyses of the sequence data showed site clustering based primarily on temperature instead of pH or sample site, suggesting that while pH, temperature, and sample site were all shown to be significant, temperature is the strongest factor driving microorganism community development. While it is important to characterize the microorganism community present, it is also important to understand how this community functions as a result of its selection. Along with looking at community composition, genomic material was tested to see if it contained mercury methylating (hgcA) or mercury reducing (merA) genes. Out of 22 samples, three of them were observed to have merA genes, while no samples had hgcA genes. These results indicate that microorganisms in Mustard and Nymph Springs may use mercury reduction. Understanding how microorganisms survive in environments with high concentrations of toxic pollutants is crucial because it can be used as a model to better understand mechanisms of resistance and the biogeochemical cycle, as well as for bioremediation and other solutions to anthropogenic problems.

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.

Bacteria employ adaptive mechanisms of mercury (Hg) tolerance to survive in environments containing elevated Hg concentrations. The potential of extracellular polysaccharides (EPS) production by bacteria as a mechanism of Hg tolerance has not been previously investigated. The objectives of this study were to determine if bacterial EPS sorb Hg, and if so does sorption provide protection against Hg toxicity. Purified EPS with different chemical compositions produced by bacterial isolates from microbial mats in French Polynesian atolls and deep-sea hydrothermal vents were assessed for Hg sorption. The data showed that EPS sorbed up to 82% of Hg from solution, that this sorption was dependent on EPS composition, and that sorption was a saturable mechanism. Hg uptake capacities ranged from 0.005 to 0.454 mmol Hg/g for the different EPS. To determine if EPS production could alter bacterial Hg tolerance, Escherichia coli K-12 strains and their EPS defective mutants were tested by the disc inhibition assay. Mercury inhibited growth in a dose-dependent manner with wild-type strains having smaller (~1 mm), but statistically significant, zones of inhibition than various mutants and this difference was related to a 2-fold decline in the amount of EPS produced by the mutants relative to cell biomass. These experiments identified colanic acid and hexosamine as Hg-binding moieties in EPS. Together these data indicate that binding of Hg to EPS affords a low level of resistance to the producing bacteria.

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.

Mercury (Hg) is a highly toxic and widely distributed heavy metal, which some Bacteria and Archaea detoxify by the reduction of ionic Hg (Hg[II]) to the elemental volatile form, Hg(0). This activity is specified by the mer operon. The mer operon of the deeply branching thermophile Thermus thermophilus HB27 encodes for, an O-acetyl-l-homoacetylserine sulfhydrylase (Oah2), a transcriptional regulator (MerR), a hypothetical protein (hp) and a mercuric reductase (MerA). Here, we show that this operon has two convergently expressed and differentially regulated promoters. An upstream promoter, P oah , controls the constitutive transcription of the entire operon and a second promoter (P mer ), located within merR, is responsive to Hg(II). In the absence of Hg(II), the transcription of merA is basal and when Hg(II) is present, merA transcription is induced. This response to Hg(II) is controlled by MerR and genetic evidence suggests that MerR acts as a repressor and activator of P mer . When the whole merR, including P mer , is removed, merA is transcribed from P oah independently of Hg(II). These results suggest that the transcriptional regulation of mer in T. thermophilus is both similar to, and different from, the well-documented regulation of proteobacterial mer systems, possibly representing an early step in the evolution of mer-operon regulation.

International audience; Objectives: This study examined the role of resistance-nodulation-cell division (RND) efflux pumps in resistance to first-generation and third-generation cephalosporins, and the potential contribution to increased virulence in two Vibrio isolates from the gut microbiota of a forage-feeder fish. Methods: Phenotypic MIC testing was performed in the presence and absence of an RND efflux pump inhibitor, phenylalanine-arginine-beta-napthylamide (PA beta N). Genomes of the two Vibrio spp. were compared to characterise RND efflux pump gene homologs. Results: The study identified 13 and 12 RND operons, respectively, in Vibrio spp. T21 and T9, with Vibrio sp. T21 containing an additional RND operon compared with other V. parahaemolyticus strains. Both the inner-membrane protein (IMP) and the membrane facilitator protein (MFP) sequences of this operon were homologous to VexD and VexC, respectively, which is an RND operon in Vibrio cholerae. More generally, the other RND proteins in these strains showed homology to RND efflux pumps characterised in Escherichia coli and Vibrio cholerae. Decreased resistance to cefoperazone and cephradine was observed in Vibrio sp. T21, and to cefoperazone and cefsulodin in Vibrio sp. T9 in the presence of Pa beta N. The RND pumps may also mediate transport of kanamycin. Conclusions: By analysing the genomes of two Vibrio spp. isolated from the mummichog fish gut, RND efflux pump-mediated resistance to first-generation and third-generation cephalosporins was discovered in these strains. This work highlights the need for further research into this unique Vibrio spp. operon and, more generally, RND efflux pumps in Vibrio spp., as Vibrio spp. often cause seafood-borne illness. (C) 2019 International Society for Antimicrobial Chemotherapy. Published by Elsevier Ltd. All rights reserved.

The emergence and spread of antibiotic-resistant pathogenic bacteria is currently one of the most serious challenges to human health. To combat this problem, it is critical to understand the processes and pathways that result in the creation of antibiotic resistance gene pools in the environment. In this study, we examined the effects of mercury (Hg) exposure on the co-selection of Hg and antibiotic-resistant bacteria that colonize the gastrointestinal tract of the mummichog (Fundulus heteroclitus), a small, estuarine fish. We examined this connection in two experimental systems: (i) a short-term laboratory exposure study where fish were fed Hg-laced food for 15 days and (ii) an examination of environmental populations from two sites with very different levels of Hg contamination. In the lab exposure study, fish muscle tissue accumulation of Hg was proportional to food Hg concentration (R 2 = 0.99; P

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.

International audience; Antibiotic resistance is a global public health issue and metal exposure can co-select for antibiotic resistance. We examined genome sequences of three multi-drug and metal resistant bacteria: one Shewanella sp., and two Vibrio spp., isolated from the gut of the mummichog fish (Fundulus heteroclitus). Our primary goal was to understand the mechanisms of co-selection. Phenotypically, the strains showed elevated resistance to arsenate, mercury, and various types of β-lactams. The genomes contained genes of public health concern including one carbapenemase (bla OXA-48). Our analyses indicate that the co-selection phenotype is mediated by chromo-somal resistance genes and cross-resistance. No evidence of co-resistance was found; most resistance genes were chromosomally located. Moreover, the identification of many efflux pump gene homologs indicates that cross-resistance and/or co-regulation may further contribute to resistance. We suggest that the mummichog gut microbiota may be a source of clinically relevant antibiotic resistance genes.

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 (Hg), one of the most toxic and widely distributed heavy metals, has a high affinity for thiol groups. Thiol groups reduce and sequester Hg. Therefore, low-molecular-weight (LMW) and protein thiols may be important cell components used in Hg resistance. To date, the role of low-molecular-weight thiols in Hg detoxification remains understudied. The mercury resistance ( mer ) operon of Thermus thermophilus suggests an evolutionary link between Hg(II) resistance and low-molecular-weight thiol metabolism. The mer operon encodes an enzyme involved in methionine biosynthesis, Oah. Challenge with Hg(II) resulted in increased expression of genes involved in the biosynthesis of multiple low-molecular-weight thiols (cysteine, homocysteine, and bacillithiol), as well as the thioredoxin system. Phenotypic analysis of gene replacement mutants indicated that Oah contributes to Hg resistance under sulfur-limiting conditions, and strains lacking bacillithiol and/or thioredoxins are more sensitive to Hg(II) than the wild type. Growth in the presence of either a thiol-oxidizing agent or a thiol-alkylating agent increased sensitivity to Hg(II). Furthermore, exposure to 3 μM Hg(II) consumed all intracellular reduced bacillithiol and cysteine. Database searches indicate that oah2 is present in all Thermus sp. mer operons. The presence of a thiol-related gene was also detected in some alphaproteobacterial mer operons, in which a glutathione reductase gene was present, supporting the role of thiols in Hg(II) detoxification. These results have led to a working model in which LMW thiols act as Hg(II)-buffering agents while Hg is reduced by MerA. IMPORTANCE The survival of microorganisms in the presence of toxic metals is central to life's sustainability. The affinity of thiol groups for toxic heavy metals drives microbe-metal interactions and modulates metal toxicity. Mercury detoxification ( mer ) genes likely originated early in microbial evolution in geothermal environments. Little is known about how mer systems interact with cellular thiol systems. Thermus spp. possess a simple mer operon in which a low-molecular-weight thiol biosynthesis gene is present, along with merR and merA . In this study, we present experimental evidence for the role of thiol systems in mercury resistance. Our data suggest that, in T. thermophilus , thiolated compounds may function side by side with mer genes to detoxify mercury. Thus, thiol systems function in consort with mer -mediated resistance to mercury, suggesting exciting new questions for future research.

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 ...

Methylmercury (MeHg), a neurotoxic substance that accumulates in aquatic food chains and poses a risk to human health, is synthesized by anaerobic microorganisms in the environment. To date, mercury (Hg) methylation has been attributed to sulfate- and iron-reducing bacteria (SRB and IRB, respectively). Here we report that a methanogen, Methanospirillum hungatei JF-1, methylated Hg in a sulfide-free medium at comparable rates, but with higher yields, than those observed for some SRB and IRB. Phylogenetic analyses showed that the concatenated orthologs of the Hg methylation proteins HgcA and HgcB from M. hungatei are closely related to those from known SRB and IRB methylators and that they cluster together with proteins from eight other methanogens, suggesting that these methanogens may also methylate Hg. Because all nine methanogens with HgcA and HgcB orthologs belong to the class Methanomicrobia , constituting the late-evolving methanogenic lineage, methanogenic Hg methylation could not be considered an ancient metabolic trait. Our results identify methanogens as a new guild of Hg-methylating microbes with a potentially important role in mineral-poor (sulfate- and iron-limited) anoxic freshwater environments.

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.

Metals are toxic to both plants and fungi, and elevated soil metal concentrations have been documented to change the structure of ectomycorrhizal communities. Mercury (Hg) is a highly toxic metal and inhibits the growth of ectomycorrhizal fungi (ECMF) in axenic culture. However, the effects of Hg on the growth of tree seedlings and the development of their ECMF communities have not been explored. In the current study, Pinus rigida seedlings were planted in soil amended with 0–366 μg g −1 Hg and incubated for 5 months. Survival and growth of P. rigida seedlings was determined, and their ECMF communities were characterized by morphotype analysis. Seedling survival declined with increasing Hg additions, although no reduction in growth was detected among surviving seedlings. The addition of 88 μg g −1 Hg to soil more than halved the total ectomycorrhizal colonization of root tips and reduced both richness and diversity of root tip morphotypes, while lower Hg additions did not significantly affect ECMF community composition relative to the no Hg control. This suggests that changes in the community of ECMF may occur in contaminated soils before any aboveground effects on surviving seedlings are noticeable, potentially altering the contribution of ECMF to the fitness of established host plants.

PurposeBy focusing on the intra‐organization dimension of corporate social responsibility (CSR), this paper aims to offer an ethnographic analysis of the way Coca‐Cola integrates its re‐branding and marketing strategies with CSR and the processes through which this strategic agenda is diffused into the company's national franchise in Israel.Design/methodology/approachThe research is based on a combined qualitative methodology of interviews with managers and employees on all levels of the organization; participant observations of formal meetings, company events and informal gatherings; and document evaluation.FindingsThis paper shows how the CSR program is purposefully and rationally designed to meet the standards of a business case approach to CSR and that, accordingly, company managers integrate it into the activities of departments and divisions such as sales, marketing, and human resources (HR). The paper further shows that the cause‐marketing and product‐branding goals underlying the global re‐branding strategy of Coca‐Cola have been mediated through employee volunteering projects that are based on the recruitment and mobilization of the physical bodies of employees not only as “bodily‐corporate‐producers”, but also as “bodily‐corporate‐ambassadors”.Research limitations/implicationsThe in‐depth empirical research that focused on a single corporation enabled the author to reach a better understanding of the intricate organizational processes involved in the implementation of CSR programs and their effects. At the same time, the research plan lacks a comparative examination of similar processes in other corporations. In this regard, this paper delineates the theoretical and empirical contours for future studies on the actual effects of implementation processes of the business case model for CSR.Originality/valueThe paper enriches a missing part in the literature regarding empirical examination of the impact of CSR at close range and at the level of individual firms.

Community involvement programs occupy centre-stage in the portfolio of many corporations who display and report upon their socially responsible performance. Focusing mainly on issues such as charity and employee volunteering, corporations remain fairly vague in reporting on the way they translate community involvement policies into concrete actions and on the social impact of their community programs. Based on first-hand observations and on-site ethnographic accounts, this study seeks to enrich extant understandings of the character and consequences of corporate involvement in communities. The study follows the diffusion of Coca-Cola’s global branding strategy and the community involvement program it recommended to the Israeli franchisee and analyzes its design and execution on the ground. The study finds a considerable gap between rhetoric of community involvement and practices of mobilizing the community to further the company’s ends. On a theoretical level, the study shows that community programs function as material performances of present-day capitalist ideology.

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.

The distribution and phylogeny of extant protein-encoding genes recovered from geochemically diverse environments can provide insight into the physical and chemical parameters that led to the origin and which constrained the evolution of a functional process. Mercuric reductase (MerA) plays an integral role in mercury (Hg) biogeochemistry by catalyzing the transformation of Hg(II) to Hg(0). Putative merA sequences were amplified from DNA extracts of microbial communities associated with mats and sulfur precipitates from physicochemically diverse Hg-containing springs in Yellowstone National Park, Wyoming, using four PCR primer sets that were designed to capture the known diversity of merA. The recovery of novel and deeply rooted MerA lineages from these habitats supports previous evidence that indicates merA originated in a thermophilic environment. Generalized linear models indicate that the distribution of putative archaeal merA lineages was constrained by a combination of pH, dissolved organic carbon, dissolved total mercury and sulfide. The models failed to identify statistically well supported trends for the distribution of putative bacterial merA lineages as a function of these or other measured environmental variables, suggesting that these lineages were either influenced by environmental parameters not considered in the present study, or the bacterial primer sets were designed to target too broad of a class of genes which may have responded differently to environmental stimuli. The widespread occurrence of merA in the geothermal environments implies a prominent role for Hg detoxification in these environments. Moreover, the differences in the distribution of the merA genes amplified with the four merA primer sets suggests that the organisms putatively engaged in this activity have evolved to occupy different ecological niches within the geothermal gradient.

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

Mercuric reductase (MerA) is central to the mercury (Hg) resistance (mer) system, catalyzing the reduction of ionic Hg to volatile Hg(0). A total of 213 merA homologues were identified in sequence databases, the majority of which belonged to microbial lineages that occupy oxic environments. merA was absent among phototrophs and in lineages that inhabit anoxic environments. Phylogenetic reconstructions of MerA indicate that (i) merA originated in a thermophilic bacterium following the divergence of the Archaea and Bacteria with a subsequent acquisition in Archaea via horizontal gene transfer (HGT), (ii) HGT of merA was rare across phylum boundaries and (iii) MerA from marine bacteria formed distinct and strongly supported lineages. Collectively, these observations suggest that a combination of redox, light and salinity conditions constrain MerA to microbial lineages that occupy environments where the most oxidized and toxic form of Hg, Hg(II), predominates. Further, the taxon-specific distribution of MerA with and without a 70 amino acid N-terminal extension may reflect intracellular levels of thiols. In conclusion, MerA likely evolved following the widespread oxygenation of the biosphere in a thermal environment and its subsequent evolution has been modulated by the interactions of Hg with the intra- and extracellular environment of the organism.

Processes leading to the bioaccumulation of methylmercury (MeHg) in northern wetlands are largely unknown. We have studied various ecological niches within a remote, acidic forested lake ecosystem in the southwestern Adirondacks, NY, to discover that mats comprised of Sphagnum moss were a hot spot for mercury (Hg) and MeHg accumulation (190.5 and 18.6 ng g⁻¹ dw, respectively). Furthermore, significantly higher potential methylation rates were measured in Sphagnum mats as compared with other sites within Sunday Lake's ecosystem. Although MPN estimates showed a low biomass of sulfate-reducing bacteria (SRB), 2.8 × 10⁴ cells mL⁻¹ in mat samples, evidence consisting of (1) a twofold stimulation of potential methylation by the addition of sulfate, (2) a significant decrease in Hg methylation in the presence of the sulfate reduction inhibitor molybdate, and (3) presence of dsrAB-like genes in mat DNA extracts, suggested that SRB were involved in Hg methylation. Sequencing of dsrB genes indicated that novel SRB, incomplete oxidizers including Desulfobulbus spp. and Desulfovibrio spp., and syntrophs dominated the sulfate-reducing guild in the Sphagnum moss mat. Sphagnum, a bryophyte dominating boreal peatlands, and its associated microbial communities appear to play an important role in the production and accumulation of MeHg in high-latitude ecosystems.

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.

In this article I analyze a multi-stakeholder process of environmental regulation. By grounding the article in the literature on regulatory capitalism and governance, I follow the career of a specific legislative process: the enactment of Israel's Deposit Law on Beverage Containers, which aims to delegate the responsibility for recycling to industry. I show that one crucial result of this process was the creation of a non-profit entity licensed to act as a compliance mechanism. This new entity enabled industry to distance itself from the responsibility of recycling, and thereby frustrated the original objective of the legislation, which was to implement the principle of “extended producer responsibility.” Furthermore, this entity, owned by commercial companies and yet acting as an environmentally friendly organization, allowed industry to promote an anti-regulatory agenda via a “civic voice.” The study moves methodologically from considering governance as an institutional structure to analyzing the process of “governancing,” through which authoritative capacities and legal responsibilities are distributed among state and non-state actors. Two key findings are that this process and its outcome (i) are premised on an ideology of civic voluntarism, which ultimately delegates environmental responsibilities to citizens; and (ii) facilitate an anti-regulatory climate that serves commercial interests.

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.