Nutrient capture in an Iowa farm pond: Insights from high-frequency observations.

Shallow constructed ponds are abundant landscape features in the midwestern United States, suggested as an edge of field best management practice (BMP) in voluntary nutrient reduction strategies. The efficacy of such features is highly uncertain, however, and previous studies have lacked sufficient temporal resolution to determine N and P removals during critical periods of transport. We utilized high-frequency in-situ measurements and flow-weighted grab sampling to determine water and nutrient budgets for a typical constructed "farm pond" in central Iowa situated within the Iowa Southern Drift Plain. Our monitoring approach yielded insight into in-stream nitrogen processing and the relative importance of transport-vs. supply-limited N delivery. Diel patterns in NO3-N observed during early Spring, prior to canopy closure, revealed that in-stream primary production and NO3-N assimilation can influence downstream N delivery in a stream with nitrate pollution (mean annual NO3-N of nearly 5 mg/L). Analysis of discharge-concentration hysteresis for NO3-N showed a shift from transport to supply limitation for NO3-N delivery over the growing season, influenced by antecedent moisture, with wet antecedent conditions leading to supply limitation. Significant NO3-N removal (64% of 19.8 kg/ha inputs) occurred within the 4.2 ha pond (230 ha watershed), but total N removal was much lower (36% removal of 22.3 kg/ha inputs). The lower total N removal highlights the importance of both particulate N and dissolved organic N and ammonia export to the N budgets of hypereutrophic small ponds. Total P removal in the pond was only 8% of 2.3 kg/ha inputs, likely due to internal loading of recent and legacy sedimentary P within the pond. High-flow events dominated N and P inputs, during which removal efficacy of the pond was significantly diminished. Poor process performance during critical moments may partially explain lower than expected water quality improvements post-BMP implementation. Accordingly, shifting hydroclimatic regimes (e.g., frequency of intense rainfall events) will impact the efficacy of small ponds and other edge of field BMPs for nutrient reduction.

Dimethylmercury Degradation by Dissolved Sulfide and Mackinawite.

Potential degradation pathways of dimethylmercury (DMHg) remain as one of the critical knowledge gaps in the marine biogeochemical cycle of mercury (Hg). Although Hg is known to be highly reactive with reduced sulfur, demethylation of DMHg in the presence of sulfide has until now remained experimentally untested. Here, we provide the first experimental support for demethylation of DMHg to monomethylmercury (MMHg) in the presence of both dissolved sulfide and mackinawite (FeS(s)m). The degradation of DMHg was shown to be pH dependent, with higher demethylation rates at pH 9 than pH 5. At room temperature and environmentally relevant DMHg to sulfide molar ratios, we observed demethylation rates up to 0.05 d-1. When comparing the number of active sites available, FeS(s)m was found to have a higher capacity to demethylate DMHg, in comparison with dissolved sulfide. Our study suggests that dissolved sulfide and FeS(s)m mediated demethylation of DMHg may act as a sink for DMHg, and a potential source of MMHg, in aquatic systems.

Emerging investigator series: methylmercury speciation and dimethylmercury production in sulfidic solutions.

Alkylated mercury species (monomethylmercury, MeHg, and dimethylmercury, DMeHg) exhibit significant bioaccumulation, and pose significant risks to ecosystems and human health. Although decades of research have been devoted to understanding MeHg formation and degradation, little is known about the DMeHg formation in aquatic systems. Here, we combine complementary experimental and computational approaches to examine MeHg speciation and DMeHg formation in sulfidic aqueous solutions, with an emphasis on the formation and decomposition of the binuclear bis(methylmercuric(ii)) sulfide complex (CH3Hg)2S. Experimental data indicate that the reaction 2CH3Hg+ + HS- ⇄ (CH3Hg)2S + H+ has a log K = 26.0 ± 0.2. Thus, the binuclear (CH3Hg)2S complex is likely to be the dominant MeHg species under high MeHg concentrations typically used in experimental investigations of MeHg degradation by sulfate-reducing bacteria (SRB). Our finding of a significant abiotic removal mechanism for MeHg in sulfidic solutions through the formation of relatively insoluble (CH3Hg)2S suggests careful reexamination of reported "oxidative demethylation" of MeHg by SRB and perhaps other obligate anaerobes. We provide evidence for slow decomposition of (CH3Hg)2S to DMeHg and HgS, with a first-order rate constant k = 1.5 ± 0.4 × 10-6 h-1. Quantum chemical calculations suggest that the reaction proceeds by a novel mechanism involving rearrangement of the (CH3Hg)2S complex facilitated by strong Hg-Hg interactions that activate a methyl group for intramolecular transfer. Predictions of DMeHg formation rates under a variety of field and laboratory conditions indicate that this pathway for DMeHg formation will be significant in laboratory experiments utilizing high MeHg concentrations, favoring (CH3Hg)2S formation. In natural systems with relatively high MeHg/[H2S]T ratios (the oxic/anoxic interface, for example), DMeHg production may be observed, and warrants further investigation.

Sulfurization of Dissolved Organic Matter Increases Hg-Sulfide-Dissolved Organic Matter Bioavailability to a Hg-Methylating Bacterium.

Reactions of dissolved organic matter (DOM) with aqueous sulfide (termed sulfurization) in anoxic environments can substantially increase DOM's reduced sulfur functional group content. Sulfurization may affect DOM-trace metal interactions, including complexation and metal-containing particle precipitation, aggregation, and dissolution. Using a diverse suite of DOM samples, we found that susceptibility to additional sulfur incorporation via reaction with aqueous sulfide increased with increasing DOM aromatic-, carbonyl-, and carboxyl-C content. The role of DOM sulfurization in enhancing Hg bioavailability for microbial methylation was evaluated under conditions typical of Hg methylation environments (µM sulfide concentrations and low Hg-to-DOM molar ratios). Under the conditions of predicted metacinnabar supersaturation, microbial Hg methylation increased with increasing DOM sulfurization, likely reflecting either effective inhibition of metacinnabar growth and aggregation or the formation of Hg(II)-DOM thiol complexes with high bioavailability. Remarkably, Hg methylation efficiencies with the most sulfurized DOM samples were similar (>85% of total Hg methylated) to that observed in the presence of l-cysteine, a ligand facilitating rapid Hg(II) biouptake and methylation. This suggests that complexes of Hg(II) with DOM thiols have similar bioavailability to Hg(II) complexes with low-molecular-weight thiols. Overall, our results are a demonstration of the importance of DOM sulfurization to trace metal and metalloid (especially mercury) fate in the environment. DOM sulfurization likely represents another link between anthropogenic sulfate enrichment and MeHg production in the environment.

Mercury methylation by novel microorganisms from new environments.

Microbial mercury (Hg) methylation transforms a toxic trace metal into the highly bioaccumulated neurotoxin methylmercury (MeHg). The lack of a genetic marker for microbial MeHg production has prevented a clear understanding of Hg-methylating organism distribution in nature. Recently, a specific gene cluster (hgcAB) was linked to Hg methylation in two bacteria.1 Here we test if the presence of hgcAB orthologues is a reliable predictor of Hg methylation capability in microorganisms, a necessary confirmation for the development of molecular probes for Hg-methylation in nature. Although hgcAB orthologues are rare among all available microbial genomes, organisms are much more phylogenetically and environmentally diverse than previously thought. By directly measuring MeHg production in several bacterial and archaeal strains encoding hgcAB, we confirmed that possessing hgcAB predicts Hg methylation capability. For the first time, we demonstrated Hg methylation in a number of species other than sulfate- (SRB) and iron- (FeRB) reducing bacteria, including methanogens, and syntrophic, acetogenic, and fermentative Firmicutes. Several of these species occupy novel environmental niches for Hg methylation, including methanogenic habitats such as rice paddies, the animal gut, and extremes of pH and salinity. Identification of these organisms as Hg methylators now links methylation to discrete gene markers in microbial communities.

Effect of dissolved organic matter source and character on microbial Hg methylation in Hg-S-DOM solutions.

Dissolved organic matter (DOM) is a key component of fate and transport models for most metals, including mercury (Hg). Utilizing a suite of diverse DOM isolates, we demonstrated that DOM character, in addition to concentration, influences inorganic Hg (Hg(II)i) bioavailability to Hg-methylating bacteria. Using a model Hg-methylating bacterium, Desulfovibrio desulfuricans ND132, we evaluated Hg-DOM-sulfide bioavailability in washed-cell assays at environmentally relevant Hg/DOM ratios (∼1-8 ng Hg/mg C) and sulfide concentrations (1-1000 µM). All tested DOM isolates significantly enhanced Hg methylation above DOM-free controls (from ∼2 to >20-fold for 20 mg C/L DOM solutions), but high molecular weight/highly aromatic DOM isolates and/or those with high sulfur content were particularly effective at enhancing Hg methylation. Because these experiments were conducted under conditions of predicted supersaturation with respect to metacinnabar (ß-HgS(s)), we attribute the DOM-dependent enhancement of Hg(II)i bioavailability to steric and specific chemical (e.g., DOM thiols) inhibition of ß-HgS(s) growth and aggregation by DOM. Experiments examining the role of DOM across a wide sulfide gradient revealed that DOM only enhances Hg methylation under fairly low sulfide conditions (≲30 µM), conditions that favor HgS nanoparticle/cluster formation relative to dissolved HgS species.

Detailed assessment of the kinetics of Hg-cell association, Hg methylation, and methylmercury degradation in several Desulfovibrio species.

The kinetics of inorganic Hg [Hg(II)(i)] association, methylation, and methylmercury (MeHg) demethylation were examined for a group of Desulfovibrio species with and without MeHg production capability. We employed a detailed method for assessing MeHg production in cultures, including careful control of medium chemistry, cell density, and growth phase, plus mass balance of Hg(II)(i) and MeHg during the assays. We tested the hypothesis that differences in Hg(II)(i) sorption and/or uptake rates drive observed differences in methylation rates among Desulfovibrio species. Hg(II)(i) associated rapidly and with high affinity to both methylating and nonmethylating species. MeHg production by Hg-methylating strains was rapid, plateauing after ∼3 h. All MeHg produced was rapidly exported. We also tested the idea that all Desulfovibrio species are capable of Hg(II)(i) methylation but that rapid demethylation masks its production, but we found this was not the case. Therefore, the underlying reason why MeHg production capability is not universal in the Desulfovibrio is not differences in Hg affinity for cells nor differences in the ability of strains to degrade MeHg. However, Hg methylation rates varied substantially between Hg-methylating Desulfovibrio species even in these controlled experiments and after normalization to cell density. Thus, biological differences may drive cross-species differences in Hg methylation rates. As part of this study, we identified four new Hg methylators (Desulfovibrio aespoeensis, D. alkalitolerans, D. psychrotolerans, and D. sulfodismutans) and four nonmethylating species (Desulfovibrio alcoholivorans, D. tunisiensis, D. carbinoliphilus, and D. piger) in our ongoing effort to generate a library of strains for Hg methylation genomics.

Dissolved organic matter enhances microbial mercury methylation under sulfidic conditions.

Dissolved organic matter (DOM) is generally thought to lower metal bioavailability in aquatic systems due to the formation of metal-DOM complexes that reduce free metal ion concentrations. However, this model may not be pertinent for metal nanoparticles, which are now understood to be ubiquitous, sometimes dominant, metal species in the environment. The influence of DOM on Hg bioavailability to microorganisms was examined under conditions (0.5-5.0 nM Hg and 2-10 µM sulfide) that favor the formation of ß-HgS(s) (metacinnabar) nanoparticles. We used the methylation of stable-isotope enriched (201)HgCl(2) by Desulfovibrio desulfuricans ND132 in short-term washed cell assays as a sensitive, environmentally significant proxy for Hg uptake. Suwannee River humic acid (SRHA) and Williams Lake hydrophobic acid (WLHPoA) substantially enhanced (2- to 38-fold) the bioavailability of Hg to ND132 over a wide range of Hg/DOM ratios (9.4 pmol/mg DOM to 9.4 nmol/mg DOM), including environmentally relevant ratios. Methylmercury (MeHg) production by ND132 increased linearly with either SRHA or WLHPoA concentration, but SRHA, a terrestrially derived DOM, was far more effective at enhancing Hg-methylation than WLHPoA, an aquatic DOM dominated by autochthonous sources. No DOM-dependent enhancement in Hg methylation was observed in Hg-DOM-sulfide solutions amended with sufficient l-cysteine to prevent ß-HgS(s) formation. We hypothesize that small HgS particles, stabilized against aggregation by DOM, are bioavailable to Hg-methylating bacteria. Our laboratory experiments provide a mechanism for the positive correlations between DOC and MeHg production observed in many aquatic sediments and wetland soils.

Rates of hexavalent chromium reduction in anoxic estuarine sediments: pH effects and the role of acid volatile sulfides.

Rates of Cr(VI) reduction were assessed in batch kinetic experiments using dilute suspensions of anoxic sediments collected from the Baltimore Harbor. Rapid and complete Cr(VI) reduction occurred on the time scale of minutes (pseudo-first-order rate constants ranged from 0.01 to 0.40 min(-1) for 1.0 g/L sediment suspensions at pH 7.0) via reaction with mineral surfaces. The reaction rate was first-order with respect to Cr(VI) concentration and proportional to the concentration of sediment acid volatile sulfides (AVS). AVS-normalized rates of Cr(VI) reduction decreased by approximately 2 orders of magnitude as the suspension pH increased from 5.0 to 8.2. The AVS-normalized rate constant was linearly correlated with the fraction of total dissolved Cr(VI) in the protonated HCrO(4)(-) form at a given pH (f(HCrO(4))((-))). The following rate law was found to describe all the experimental data: d[Cr(VI)]/dt = -k[AVS]f(HCrO(4))((-))[Cr(VI)](T), where the AVS- and pH-independent rate constant k = 3.14(+/-0.75) x 10(4) M(-1) min(-1). The results of this study suggest the importance of reactions between Cr(VI) and solid phase AVS constituents in controlling Cr(VI) fate in anoxic estuarine sediments, and indicate that Cr(VI) is unlikely to be a toxicological stressor in AVS-containing sediments.

Chromium occurrence and speciation in Baltimore harbor sediments and porewater, Baltimore, Maryland, USA.

Industrial activities in the Baltimore Harbor, Baltimore, Maryland, USA, have resulted in widespread chromium contamination of sediments. A comprehensive analysis of Cr speciation in sediment and porewater collected from 22 locations in the Baltimore Harbor was completed to understand Cr bioavailability and probability of toxicity due to Cr in sediments. The analysis employed a reverse-phase ion-pair high-performance liquid chromatography-inductively coupled plasma-mass spectrometry (HPLC-ICP-MS) method. Sub-microgram-per-liter determination of Cr(III) and Cr(VI) in environmental samples was found, with method validation revealing broad method applicability of HPLC-ICP-MS to a wide range of sample types. The major limitation of the method was poor Cr species separation in high ionic strength solutions (greater than 0.1 M NaCl). Total Cr concentrations in Baltimore Harbor sediments ranged from 2.5 to 1,050 mg/kg with 11 of the 22 sites containing total Cr in excess of the 370 mg/ kg effects range-median (ER-M) sediment quality guideline. The Cr(VI) concentrations in sediments, however, were markedly lower, ranging from 0.10 to 0.38 mg/kg with Cr(VI) not detected in 14 of the 22 stations. Porewater concentrations, both for total Cr and Cr(VI), were quite low, with total Cr ranging from 0.20 to 2.16 microg/L and Cr(VI) ranging from 0.73 to 1.17 microg/L. The Cr(VI)-reducing capacity of the sediments, based on a sediment-spiking experiment, was found to be strongly correlated with the acid volatile sulfides content of the sediment. Overall, our results provide field validation of the hypothesis that Cr(VI) will not persist in sediments with excess acid volatile sulfides. Given the low concentrations of Cr(VI) in sediment and porewater, it appears unlikely that Cr in Baltimore Harbor sediments contributes appreciably to previously observed sediment toxicity.