Global survey of hgcA-carrying genomes in marine and freshwater sediments: Insights into mercury methylation processes.

Mercury (Hg) methylation is a microbially mediated process that produces methylmercury (MeHg), a bioaccumulative neurotoxin. A highly conserved gene pair, hgcAB, is required for Hg methylation, which provides a basis for identifying Hg methylators and evaluating their genomic composition. In this study, we conducted a large-scale omics analysis in which 281 metagenomic freshwater and marine sediment samples from 46 geographic locations across the globe were queried. Specific objectives were to examine the prevalence of Hg methylators, to identify horizontal gene transfer (HGT) events involving hgcAB within Hg methylator communities, and to identify associations between hgcAB and microbial biochemical functions/genes. Hg methylators from the phyla Desulfobacterota and Bacteroidota were dominant in both freshwater and marine sediments while Firmicutes and methanogens belonging to Euryarchaeota were identified only in freshwater sediments. Novel Hg methylators were found in the Phycisphaerae and Planctomycetia classes within the phylum Planctomycetota, including potential hgcA-carrying anammox metagenome-assembled genomes (MAGs) from Candidatus Brocadiia. HGT of hgcA and hgcB were identified in both freshwater and marine methylator communities. Spearman's correlation analysis of methylator genomes suggested that in addition to sulfide, thiosulfate, sulfite, and ammonia may be important parameters for Hg methylation processes in sediments. Overall, our results indicated that the biochemical drivers of Hg methylation vary between marine and freshwater sites, lending insight into the influence of environmental perturbances, such as a changing climate, on Hg methylation processes.

Global stress response in Pseudomonas aeruginosa upon malonate utilization.

Versatility in carbon source utilization assists Pseudomonas aeruginosa in its adaptation to various niches. Recently, we characterized the role of malonate, an understudied carbon source, in quorum sensing regulation, antibiotic resistance, and virulence factor production in P. aeruginosa . These results indicate that global responses to malonate metabolism remain to be uncovered. We leveraged a publicly available metabolomic dataset on human airway and found malonate to be as abundant as glycerol, a common airway metabolite and carbon source for P. aeruginosa . Here, we explored and compared adaptations of P. aeruginosa UCBPP-PA14 (PA14) in response to malonate or glycerol as a sole carbon source using transcriptomics and phenotypic assays. Malonate utilization activated glyoxylate and methylcitrate cycles and induced several stress responses, including oxidative, anaerobic, and metal stress responses associated with increases in intracellular aluminum and strontium. Some induced genes were required for optimal growth of P. aeruginosa in malonate. To assess the conservation of malonate-associated responses among P. aeruginosa strains, we compared our findings in strain PA14 with other lab strains and cystic fibrosis isolates of P. aeruginosa . Most strains grew on malonate as a sole carbon source as efficiently as or better than glycerol. While not all responses to malonate were conserved among strains, formation of biomineralized biofilm-like aggregates, increased tolerance to kanamycin, and increased susceptibility to norfloxacin were the most frequently observed phenotypes. Our findings reveal global remodeling of P. aeruginosa gene expression during its growth on malonate as a sole carbon source that is accompanied by several important phenotypic changes. These findings add to accumulating literature highlighting the role of different carbon sources in the physiology of P. aeruginosa and its niche adaptation. Importance: Pseudomonas aeruginosa is a notorious pathogen that causes local and systemic infections in immunocompromised individuals. Different carbon sources can uniquely modulate metabolic and virulence pathways in P. aeruginosa , highlighting the importance of the environment that the pathogen occupies. In this work, we used a combination of transcriptomic analysis and phenotypic assays to determine how malonate utilization impacts P. aeruginosa, as recent evidence indicates this carbon source may be relevant to certain niches associated within the human host. We found that malonate utilization can induce global stress responses, alter metabolic circuits, and influence various phenotypes of P. aeruginosa that could influence host colonization. Investigating the metabolism of malonate provides insight into P. aeruginosa adaptations to specific niches where this substrate is abundant, and how it can be leveraged in the development of much-needed antimicrobial agents or identification of new therapeutic targets of this difficult-to-eradicate pathogen.

Non-mercury methylating microbial taxa are integral to understanding links between mercury methylation and elemental cycles in marine and freshwater sediments.

The goal of this study was to explore the role of non-mercury (Hg) methylating taxa in mercury methylation and to identify potential links between elemental cycles and Hg methylation. Statistical approaches were utilized to investigate the microbial community and biochemical functions in relation to methylmercury (MeHg) concentrations in marine and freshwater sediments. Sediments were collected from the methylation zone (top 15 cm) in four Hg-contaminated sites. Both abiotic (e.g., sulfate, sulfide, iron, salinity, total organic matter, etc.) and biotic factors (e.g., hgcA, abundances of methylating and non-methylating taxa) were quantified. Random forest and stepwise regression were performed to assess whether non-methylating taxa were significantly associated with MeHg concentration. Co-occurrence and functional network analyses were constructed to explore associations between taxa by examining microbial community structure, composition, and biochemical functions across sites. Regression analysis showed that approximately 80% of the variability in sediment MeHg concentration was predicted by total mercury concentration, the abundances of Hg methylating taxa, and the abundances of the non-Hg methylating taxa. The co-occurrence networks identified Paludibacteraceae and Syntrophorhabdaceae as keystone non Hg methylating taxa in multiple sites, indicating the potential for syntrophic interactions with Hg methylators. Strong associations were also observed between methanogens and sulfate-reducing bacteria, which were likely symbiotic associations. The functional network results suggested that non-Hg methylating taxa play important roles in sulfur respiration, nitrogen respiration, and the carbon metabolism-related functions methylotrophy, methanotrophy, and chemoheterotrophy. Interestingly, keystone functions varied by site and did not involve carbon- and sulfur-related functions only. Our findings highlight associations between methylating and non-methylating taxa and sulfur, carbon, and nitrogen cycles in sediment methylation zones, with implications for predicting and understanding the impact of climate and land/sea use changes on Hg methylation.

Mobility and bioaccessibility of arsenic (As) bound to titanium dioxide (TiO2) water treatment residuals (WTRs).

This work systematically describes arsenic mobility and potential bioaccessibility of arsenic-enriched titanium dioxide water treatment residuals (TiO2 WTRs) by employing a suite of wet chemical experiments and spectroscopic measurements. Specifically, Environmental Protection Agency (EPA) digestion method 3051a indicated <3% of total arsenic in the solid phase was released, and arsenic assessed by EPA method 1340 for bioaccessibility was below detection limits. A novel finding is while the arsenic appeared to be stable under highly acidic digestion conditions, it is in fact highly mobile when exposed to simple phosphate solutions. On average, 55% of arsenic was extracted from all samples during a 50-day replenishment study. This was equivalent to 169 mg kg-1 arsenic released from the solid phase. Macroscopic desorption experiments indicated arsenic likely formed inner-sphere bonds with the TiO2 particles present in the samples. This was confirmed with X-ray absorption spectroscopy (XAS), where an interatomic distance of 3.32 Å and a coordination number (CN) of 1.79 titanium atoms were determined. This translates to a configuration of arsenic on TiO2 surfaces as a bidentate binuclear inner-sphere complex. Thus, both macroscopic and spectroscopic data are in agreement. During incubation experiments, arsenic(V) was actively reduced to arsenic(III); the amount of arsenic(III) in solution varied from 8 to 38% of total dissolved arsenic. Lastly, elevated concentrations and mobility of vanadium in these systems merit further investigation. The high mobility of arsenic and its potential for reduction when reintroduced into the environment, particularly in agriculturally important areas, presents an important risk when waste products are not properly managed.

Environmental Impacts of Coal Combustion Residuals: Current Understanding and Future Perspectives.

On-site solid-waste impoundments, landfills, and receiving water bodies have served as long-term disposal sites for coal combustion residuals (CCRs) across the United States for decades and collectively contain billions of tons of CCR material. CCR components include fine particulate material, minerals, and trace elements such as mercury, arsenic, selenium, lead, etc., which can have deleterious effects on ecosystem functioning and public health. Effects on communities can occur through consumption of drinking water, fish, and other aquatic organisms. The structural failure of impoundments, water infiltration, leakage from impoundments due to poor construction and monitoring, and CCR effluent discharges to water bodies have in the past resulted in harmful environmental impacts. Moreover, the risks posed by CCRs are present to this day, as coal continues to account for 11% of the energy production in the United States. In this Critical Review, the legacy of CCR disposal and the concomitant risks posed to public health and ecosystems are assessed. The resiliency of CCR disposal sites in the context of increased frequency and intensity of storm events and other hazards, such as floods and earthquakes, is also evaluated. We discuss the current state of knowledge on the environmental fate of CCR-derived elements, as well as advances in and limitations of analytical tools, which can improve the current understanding of CCR environmental impacts in order to mitigate the associated risks. An assessment of the 2015 Coal Ash Final Rule is also presented, along with needs to improve monitoring of CCR disposal sites and regulatory enforcement.

Impacts of Sediment Particle Grain Size and Mercury Speciation on Mercury Bioavailability Potential.

Particle-specific properties, including size and chemical speciation, affect the reactivity of mercury (Hg) in natural systems (e.g., dissolution or methylation). Here, terrestrial, river, and marine sediments were size-fractionated and characterized to correlate particle-specific properties of Hg-bearing solids with their bioavailability potential and measured biomethylation. Marine sediments contained ∼20-50% of the total Hg in the <0.5 µm size fraction, compared to only 0.5 and 3.0% in this size fraction for terrestrial and river sediments, respectively. X-ray absorption spectroscopy (XAS) analysis indicated that metacinnabar (ß-HgS) was the main mercury species in a marine sediment, whereas organic Hg-thiol (Hg(SR)2) was the main mercury species in a terrestrial sediment. Single-particle inductively coupled plasma time-of-flight mass spectrometry analysis of the marine sediment suggests that half of the Hg in the <0.5 µm size fraction existed as individual nanoparticles, which were ß-HgS based on XAS analyses. Glutathione-extractable mercury was higher for samples containing Hg(SR)2 species than ß-HgS species and correlated well with the amount of Hg biomethylation. This particle-scale understanding of how Hg speciation and particle size affect mercury bioavailability potential helps explain the heterogeneity in Hg methylation in natural sediments.

Arsenic Speciation in Bituminous Coal Fly Ash and Transformations in Response to Redox Conditions.

The risk of the mobilization of coal ash into the environment has highlighted the need for the assessment of the environmental behavior of coal ash, particularly with respect to toxic trace elements such as arsenic (As). Here, we examined As speciation in coal fly ash samples and transformations in response to aquatic redox conditions. X-ray absorption spectroscopy indicated that 92-97% of total As occurred as As(V), with the remainder present as As(III). Major As-bearing hosts in unamended ashes were glass, iron (oxyhydr)oxides, and calcium arsenate. Oxic leaching resulted in immediate As mobilization to the aqueous phase, reprecipitation of As-iron ferrihydrite, and As adsorption to mineral surfaces. Under anoxic conditions, the (reductive) dissolution of As-bearing phases such as iron ferrihydrite resulted in increased dissolved As compared to oxic conditions and reprecipitation of iron arsenate. Overall, As in coal ash is not environmentally stable and can participate in local biogeochemical cycles.

Legacy source of mercury in an urban stream-wetland ecosystem in central North Carolina, USA.

In the United States, aquatic mercury contamination originates from point and non-point sources to watersheds. Here, we studied the contribution of mercury in urban runoff derived from historically contaminated soils and the subsequent production of methylmercury in a stream-wetland complex (Durham, North Carolina), the receiving water of this runoff. Our results demonstrated that the mercury originated from the leachate of grass-covered athletic fields. A fraction of mercury in this soil existed as phenylmercury, suggesting that mercurial anti-fungal compounds were historically applied to this soil. Further downstream in the anaerobic sediments of the stream-wetland complex, a fraction (up to 9%) of mercury was converted to methylmercury, the bioaccumulative form of the metal. Importantly, the concentrations of total mercury and methylmercury were reduced to background levels within the stream-wetland complex. Overall, this work provides an example of a legacy source of mercury that should be considered in urban watershed models and watershed management.

Environmental impacts of the Tennessee Valley Authority Kingston coal ash spill. 1. Source apportionment using mercury stable isotopes.

Mercury stable isotope abundances were used to trace transport of Hg-impacted river sediment near a coal ash spill at Harriman, Tennessee, USA. δ(202)Hg values for Kingston coal ash released into the Emory River in 2008 are significantly negative (-1.78 ± 0.35‰), whereas sediments of the Clinch River, into which the Emory River flows, are contaminated by an additional Hg source (potentially from the Y-12 complex near Oak Ridge, Tennessee) with near-zero values (-0.23 ± 0.16‰). Nominally uncontaminated Emory River sediments (12 miles upstream from the Emory-Clinch confluence) have intermediate values (-1.17 ± 0.13‰) and contain lower Hg concentrations. Emory River mile 10 sediments, possibly impacted by an old paper mill has δ(202)Hg values of -0.47 ± 0.04‰. A mixing model, using δ(202)Hg values and Hg concentrations, yielded estimates of the relative contributions of coal ash, Clinch River, and Emory River sediments for a suite of 71 sediment samples taken over a 30 month time period from 13 locations. Emory River samples, with two exceptions, are unaffected by Clinch River sediment, despite occasional upstream flow from the Clinch River. As expected, Clinch River sediment below its confluence with the Emory River are affected by Kingston coal ash; however, the relative contribution of the coal ash varies among sampling sites.

Environmental impacts of the Tennessee Valley Authority Kingston coal ash spill. 2. Effect of coal ash on methylmercury in historically contaminated river sediments.

The Tennessee Valley Authority Kingston coal ash spill in December 2008 deposited approximately 4.1 million m(3) of fly ash and bottom ash into the Emory and Clinch River system (Harriman, Tennessee, U.S.A.). The objective of this study was to investigate the impact of the ash on surface water and sediment quality over an eighteen month period after the spill, with a specific focus on mercury and methylmercury in sediments. Our results indicated that surface water quality was not impaired with respect to total mercury concentrations. However, in the sediments of the Emory River near the coal ash spill, total mercury concentrations were 3- to 4-times greater than sediments several miles upstream of the ash spill. Similarly, methylmercury content in the Emory and Clinch River sediments near the ash spill were slightly elevated (up to a factor of 3) at certain locations compared to upstream sediments. Up to 2% of the total mercury in sediments containing coal ash was present as methylmercury. Mercury isotope composition and sediment geochemical data suggested that elevated methylmercury concentrations occurred in regions where native sediments were mixed with coal ash (e.g., less than 28% as coal ash in the Emory River). This coal ash may have provided substrates (such as sulfate) that stimulated biomethylation of mercury. The production of methylmercury in these areas is a concern because this neurotoxic organomercury compound can be highly bioaccumulative. Future risk assessments of coal ash spills should consider not only the leaching potential of mercury from the wastes but also the potential for methylmercury production in receiving waters.

Long-term transformation and fate of manufactured ag nanoparticles in a simulated large scale freshwater emergent wetland.

Transformations and long-term fate of engineered nanomaterials must be measured in realistic complex natural systems to accurately assess the risks that they may pose. Here, we determine the long-term behavior of poly(vinylpyrrolidone)-coated silver nanoparticles (AgNPs) in freshwater mesocosms simulating an emergent wetland environment. AgNPs were either applied to the water column or to the terrestrial soils. The distribution of silver among water, solids, and biota, and Ag speciation in soils and sediment was determined 18 months after dosing. Most (70 wt %) of the added Ag resided in the soils and sediments, and largely remained in the compartment in which they were dosed. However, some movement between soil and sediment was observed. Movement of AgNPs from terrestrial soils to sediments was more facile than from sediments to soils, suggesting that erosion and runoff is a potential pathway for AgNPs to enter waterways. The AgNPs in terrestrial soils were transformed to Ag(2)S (~52%), whereas AgNPs in the subaquatic sediment were present as Ag(2)S (55%) and Ag-sulfhydryl compounds (27%). Despite significant sulfidation of the AgNPs, a fraction of the added Ag resided in the terrestrial plant biomass (~3 wt % for the terrestrially dosed mesocosm), and relatively high body burdens of Ag (0.5-3.3 µg Ag/g wet weight) were found in mosquito fish and chironomids in both mesocosms. Thus, Ag from the NPs remained bioavailable even after partial sulfidation and when water column total Ag concentrations are low (<0.002 mg/L).

Effects of humic substances on precipitation and aggregation of zinc sulfide nanoparticles.

Nanoparticulate metal sulfides such as ZnS can influence the transport and bioavailability of pollutant metals in anaerobic environments. The aim of this work was to investigate how the composition of dissolved natural organic matter (NOM) influences the stability of zinc sulfide nanoparticles as they nucleate and aggregate in water with dissolved NOM. We compared NOM fractions that were isolated from several surface waters and represented a range of characteristics including molecular weight, type of carbon, and ligand density. Dynamic light scattering was employed to monitor the growth and aggregation of Zn-S-NOM nanoparticles in supersaturated solutions containing dissolved aquatic humic substances. The NOM was observed to reduce particle growth rates, depending on solution variables such as type and concentration of NOM, monovalent electrolyte concentration, and pH. The rates of growth increased with increasing ionic strength, indicating that observed growth rates primarily represented aggregation of charged Zn-S-NOM particles. Furthermore, the observed rates decreased with increasing molecular weight and aromatic content of the NOM fractions, while carboxylate and reduced sulfur content had little effect. Differences between NOM were likely due to properties that increased electrosteric hindrances for aggregation. Overall, results of this study suggest that the composition and source of NOM are key factors that contribute to the stabilization and persistence of zinc sulfide nanoparticles in the aquatic environment.

Environmental impacts of the coal ash spill in Kingston, Tennessee: an 18-month survey.

An 18 month investigation of the environmental impacts of the Tennessee Valley Authority (TVA) coal ash spill in Kingston, Tennessee combined with leaching experiments on the spilled TVA coal ash have revealed that leachable coal ash contaminants (LCACs), particularly arsenic, selenium, boron, strontium, and barium, have different effects on the quality of impacted environments. While LCACs levels in the downstream river water are relatively low and below the EPA drinking water and ecological thresholds, elevated levels were found in surface water with restricted water exchange and in pore water extracted from the river sediments downstream from the spill. The high concentration of arsenic (up to 2000 µg/L) is associated with some degree of anoxic conditions and predominance of the reduced arsenic species (arsenite) in the pore waters. Laboratory leaching simulations show that the pH and ash/water ratio control the LCACs' abundance and geochemical composition of the impacted water. These results have important implications for the prediction of the fate and migration of LCACs in the environment, particularly for the storage of coal combustion residues (CCRs) in holding ponds and landfills, and any potential CCRs effluents leakage into lakes, rivers, and other aquatic systems.

Survey of the potential environmental and health impacts in the immediate aftermath of the coal ash spill in Kingston, Tennessee.

An investigation of the potential environmental and health impacts in the immediate aftermath of one of the largest coal ash spills in U.S. history at the Tennessee Valley Authority (TVA) Kingston coal-burning power plant has revealed three major findings. First the surface release of coal ash with high levels of toxic elements (As = 75 mg/kg; Hg = 150 microg/kg) and radioactivity (226Ra + 228Ra = 8 pCi/g) to the environment has the potential to generate resuspended ambient fine particles (< 10 microm) containing these toxics into the atmosphere that may pose a health risk to local communities. Second, leaching of contaminants from the coal ash caused contamination of surface waters in areas of restricted water exchange, but only trace levels were found in the downstream Emory and Clinch Rivers due to river dilution. Third, the accumulation of Hg- and As-rich coal ash in river sediments has the potential to have an impact on the ecological system in the downstream rivers by fish poisoning and methylmercury formation in anaerobic river sediments.

Precipitation of mercuric sulfide nanoparticles in NOM-containing water: implications for the natural environment.

Speciation of mercury(II) in the aquatic environment and coordination to natural organic matter (NOM) and sulfides governs the bioavailability and mobility of mercury in water and sediment. While previous studies on aqueous Hg(II) speciation have focused on competitive binding of dissolved species, the purpose of this study was to explore the potential for HgS nanoparticles that coprecipitate with NOM in solution. Dynamic light scattering was used to monitor the size of HgS colloids growing over time. The results indicated that humic substances decreased observed growth rates of particles and stabilized aggregates smaller than 0.2 microm for at least 8 h. Thiol-containing organic acids such as cysteine and thioglycolate also decreased growth of HgS particles. Growth rates were also monitored as a function of monovalent electrolyte concentration, humic type, and humic concentration. HgS particles that formed in the presence of humics and thiolates were able to pass through conventional filters (<0.2 micro/m) and appeared to consist of aggregates of nanocrystals in TEM images. Furthermore, 96% of HgS aggregates were removed from aqueous suspension when exposed to octanol, indicating that the particles could be incorrectly identified as dissolved complexes (e.g., HgS(0)(aq)) in bioavailability models. Hg speciation calculations were conducted to consider lower Hg concentrations observed in sediment porewater. While the calculations depended on Hg binding constants that can vary by orders of magnitude, the results indicated that HgS(s) could be oversaturated in filtered porewater, particularly at low dissolved sulfide levels (micromolar or lower). These insights suggest that nanoparticulate HgS can exist in surface waters and porewater of contaminated sediments as a result of kinetically hindered aggregation/precipitation reactions. Further studies are neededto addressthe importance of nanoscale HgS particles for governing the reactivity and bioavailability of mercury in the environment.