Mercury methylation by planktonic and biofilm cultures of Desulfovibrio desulfuricans

While biofilms are now known to be the predominant form of microbial growth in nature, very little is yet known about their role in environmental mercury (Hg) methylation. Findings of Hg methylation in periphyton communities have indicated the importance of investigating how environmental biofilms affect Hg methylation, as periphyton can be the base of the food webs in aquatic ecosystems. Chemical speciation influences the microbial uptake and methylation of inorganic Hg by planktonic cultures of sulfate-reducing bacteria; however, the effect of speciation on Hg methylation by biofilm cultures of these organisms has previously not been studied. In the present study, Hg methylation rates in biofilm and planktonic cultures of two isolates of Desulfovibrio desulfuricans from a coastal wetland were compared. Notably, the specific Hg methylation rate found was approximately an order of magnitude higher (0.0018 vs. 0.0002 attomol cell(-1) day(-1)) in biofilm cells than in planktonic cells, suggesting an important role for environmental biofilms in Hg methylation. To investigate the role of chemical speciation of Hg, experiments were conducted at two levels of sulfide. Both biofilm and planktonic cultures produced methylmercury at roughly twice the rate at low sulfide, when HgS(0)(aq), rather than HgHS2-, was the dominant Hg species. This indicates that the presence of a biofilm does not alter the relative availability of the dominant Hg species in sulfidic medium, in accordance with our previous studies of Hg uptake by Escherichia coli along a chloride gradient.     

The influence of light/dark cycle at low light frequency on the desulfurization by a photosynthetic microorganism

H2S dissolved in water can be converted to elementary sulfur or sulfate by the photosynthetic bacterium Chlorobium thiosulfatophilum. The effects of the light/dark cycle on cell growth and the rate of sulfide removal were investigated to develop an appropriate fermentation strategy. Dark fermentation was also studied without addition of H2S and CO2 as electron and carbon sources. Average specific growth rates of bacterial cultures with a continuous supply of H2S and CO2 both in light and dark conditions were occurred in the range of 0.008 to 0.009 h(-1), indicating little dependence on the light/dark cycle, but about 25% of the growth rate that was occurred only in the presence of light. Average H2S removal capacities for cultures grown under the light/dark cycles of 14/10 , 12/12 , and 9/15 h, respectively, with a continuous supply of feed gases, were 0.08, 0.07, and 0.04 micromol H2S.min(-1)/mg protein.l(-1) in the dark, and was slightly less than those in the light. H2S removal capacity with variation of the light/dark cycle was about 30-60% of that obtained in the continuously illuminated cultures. ATP concentration in the dark decreased from 0.43 to 0.37 mg ATP.mg protein(-1) as the daily dark duration decreased from 15 to 10 h. The production rate for lactic acid from a culture grown without a supply of mixtures of H2S and CO2 gases was 0.218 g lactic acid.l(-1).h(-1), much more than that grown with a supply of feed gas mixtures. Time-averaged concentrations of lactic acid produced overall during the light and dark periods were 13.7 g lactic acid.l(-1) during the light/dark cycle of 14/10 h without a supply of feed gas, and 3.1 and 2.4 g lactic acid.l(-1) during the cycles of 9/15 and 14/10 h, respectively, with a supply of feed gas.     

Photoinduced oxidation of Hg0(aq) in the waters from the St. Lawrence estuary

The oxidation of volatile aqueous Hg0 in aquatic systems may be important in decreasing the fluxes of Hg out of the water column. Using incubations of natural samples from the St. Lawrence River, we examined some of the parameters that control this oxidation. Hg0 was found to be chiefly mediated by UV radiation since (i) "dark" oxidation was not found to be statistically significant; (ii) visible light induced a significant but slow photooxidation (k = 0.09 h(-1)); and (iii) visible + UV radiation led to a faster photooxidation (k = 0.6-0.7 h(-1)), mainly because of UV-A induced reactions. Doubling UV irradiation did not increase the reaction rate of Hg0 photooxidation in natural water samples, indicating that some factor other than photon flux was rate limiting and suggesting that the reaction involves intermediate photoproduced oxidant(s). The addition of methanol, a *OH scavenger, decreased Hg photooxidation rates by 25% in brackish waters and by 19% in artificial saline water containing semiquinones, indicating that *OH may be partly responsible for Hg0 oxidation. Photooxidation rates were not affected by oxygen concentrations and did not decrease when samples were heat-sterilized, treated with chloroform, or filtered prior to exposure to light.     

Factors affecting leaf starch levels in some temperate grasses

Leaf starch was measured, by a specific enzyme method, in Lolium sp., Festuca arundinacea and Dactylis glomerata grown in the field and in contrasting temperatures of two controlled environments. Starch levels of over 4% of the dry weight and 50% of the total hot-water-soluble carbohydrate (TSC) were recorded. Levels differed significantly between and within species, but populations of each species from N. Europe amd Mediterranean regions were not consistently different. There was more starch in cool than warm conditions and it increased during light and decreased during dark periods. However, there was no consistent temperature effect on starch accumulation during 7 h light in several L. perenne plants. D. glomerata leaves contained more starch (over 4 %) but less TSC than L. perenne.     

Cobalt limitation of growth and mercury methylation in sulfate-reducing bacteria

Sulfate-reducing bacteria (SRB) have been identified as the primary organisms responsible for monomethylmercury (MeHg) production in aquatic environments, but little is known of the physiologyand biochemistry of mercury(Hg) methylation. Corrinoid compounds have been implicated in enzymatic Hg methylation, although recent experiments with a vitamin B12 inhibitor indicated that incomplete-oxidizing SRB likely do not use a corrinoid-enzyme for Hg methylation, whereas experiments with complete-oxidizing SRB were inconclusive due to overall growth limitation. Here we explore the role of corrinoid-containing methyltransferases, which contain a cobalt-reactive center, in Hg methylation. To this end, we performed cobalt-limitation experiments on two SRB strains: Desulfococcus multivorans, a complete-oxidizer that uses the acetyl-CoA pathway for major carbon metabolism, and Desulfovibrio africanus, an incomplete-oxidizer that does not contain the acetyl-CoA pathway. Cultures of D. multivorans grown with no direct addition of Co or B12 became cobalt-limited and produced 3 times less MeHg per cell than control cultures. Differences in growth rate and Hg bioavailability do not account for this large decrease in MeHg production upon Co limitation. In contrast, the growth and Hg methylation rates of D. africanus cultures remained nearly constant regardless of the inorganic cobalt and vitamin B12 concentrations in the medium. These results are consistent with mercury being methylated by different pathways in the two strains: catalyzed by a B12-containing methyltransferase in D. multivorans and a B12-independent methyltransferase in D. africanus. If complete-oxidizing SRB like D. multivorans account for the bulk of MeHg production in coastal sediments as reported, the ambient Co concentration and speciation may control the rate of Hg methylation.     

Methanogens: principal methylators of mercury in lake periphyton

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.     

New technique for quantification of elemental Hg in mine wastes and its implications for mercury evasion into the atmosphere

Mercury in the environment is of prime concern to both ecosystem and human health. Determination of the molecular-level speciation of Hg in soils and mine wastes is important for understanding its sequestration, mobility, and availability for methylation. Extended X-ray absorption fine structure (EXAFS) spectroscopy carried out under ambient P-T conditions has been used in a number of past studies to determine Hg speciation in complex mine wastes and associated soils. However, this approach cannot detect elemental (liquid) mercury in Hg-polluted soils and sediments due to the significant structural disorder of liquid Hg at ambient-temperature. A new sample preparation protocol involving slow cooling through the crystallization temperature of Hg(0) (234 K) results in its transformation to crystalline α-Hg(0). The presence and proportion of Hg(0), relative to other crystalline Hg-bearing phases, in samples prepared in this way can be quantified by low-temperature (77 K) EXAFS spectroscopy. Using this approach, we have determined the relative concentrations of liquid Hg(0) in Hg mine wastes from several sites in the California Coast Range and have found that they correlate well with measured fluxes of gaseous Hg released during light and dark exposure of the same samples, with higher evasion ratios from samples containing higher concentrations of liquid Hg(0). Two different linear relationships are observed in plots of the ratio of Hg emission under light and dark conditions vs % Hg(0), corresponding to silica-carbonate- and hot springs-type Hg deposits, with the hot springs-type samples exhibiting higher evasion fluxes than silica-carbonate type samples at similar Hg(0) concentrations. Our findings help explain significant differences in Hg evasion data for different mine sites in the California Coast Range.     

粪肠球菌Z096对副溶血弧菌生物被膜及群体感应的抑制作用

为了研究粪肠球菌Z096对副溶血弧菌生物被膜和群体感应（quorum sensing,QS）系统的抑制作用,采用竞争、清除、排阻三种方式模拟Z096与副溶血弧菌在微菌落环境中的相互作用,并进一步探究了Z096的提取物（Z096-E）对副溶血弧菌生物被膜形成、成熟生物被膜清除、细胞表面疏水性、自聚性、QS信号分子AI-2活性、群集泳动能力以及胞外多糖和蛋白合成的影响。结果表明:Z096可通过竞争、清除、排阻的方式与副溶血弧菌相互作用,降低浮游和生物被膜状态的副溶血弧菌细胞数量,干扰副溶血弧菌在载体表面的粘附,且Z096-E能够显著抑制副溶血弧菌生物被膜形成,有效清除成熟生物被膜,1.6 mg/mL的Z096-E处理12 h,副溶血弧菌生物被膜抑制率为70.43%,代谢活性减少率为84.15%;12.8 mg/mL的Z096-E处理副溶血弧菌成熟生物被膜4 h,生物被膜清除率为58.21%,代谢活性减少率为69.84%。而且1.6 mg/mL的Z096-E对副溶血弧菌群集和泳动能力、细胞表面疏水性和自聚性、胞外多糖和蛋白合成的抑制率分别为47.26%、53.56%、63.37%、89.38%、77.65%和51.91%,抑制效果具有浓度依赖性。此外,Z096-E可使副溶血弧菌QS信号分子AI-2活性减弱,表明Z096-E是一种AI-2类群体感应抑制剂,其可通过干扰QS系统,从而影响副溶血弧菌的生理特性。因此,本研究发现了一株能够抑制副溶血弧菌生物被膜的乳酸菌,其提取物Z096-E能作为一种防控副溶血弧菌生物被膜的新型乳酸菌生物制剂,这对消除致病菌生物被膜污染以及开发新型抗菌剂具有积极的作用。     

Growth dilution of metals in microalgal biofilms

Despite the key role microalgae play in introducing toxicants into aquatic food webs, little is known about the effects of environmental factors on metal accumulation by these primary producers. Environmental factors such as light and nutrients alter growth rates and may consequently influence metal concentrations in microalgae through growth dilution. Laboratory experiments suggested that metal uptake and elimination by microalgal biofilms were gradual enough to enable dilution of metals within the biofilms by photosynthetically accrued carbon, and a simple kinetic model of metal accumulation predicted significant variation in metal content due to growth dilution over the natural range of microalgal growth rates. The ratio of metal uptake to carbon uptake by microalgal biofilms decreased exponentially with increasing light in short-term laboratory experiments because photosynthesis was much more sensitive to a light gradient than was metal uptake. The effect of light on biofilm metal concentrations was confirmed in situ with a long-term experiment in which experimental shading of biofilms in a metal-contaminated stream decreased biofilm growth rates and caused a 3x increase in biofilm concentrations of twelve metals, including methylmercury. Slow growth at the primary producer level is a likely contributor to higher biotic metal concentrations in shaded, oligotrophic, or cold ecosystems.     

冷诱导金黄色葡萄球菌环境胁迫耐受性及生物被膜控制

为研究金黄色葡萄球菌经冷诱导后对其他环境胁迫耐受性的变化,将耐冷冻能力较强的分离株S.aureus W3与标准菌株S.aureus CICC10201分别在4 ℃下冷胁迫5 h后,测定其在高温、酸碱、氯化钠、过氧化氢、酒精的胁迫下存活率变化。同时对分离株S.aureus W3的生物被膜形成及控制进行了研究。结果表明,经-20 ℃冷藏8周后,S.aureus W3的存活率显著高于S.aureus CICC10201(p<0.05)。经冷诱导后,S.aureus W3在45~60 ℃环境下存活率提升至18.5%。冷诱导使金黄色葡萄球菌对酸敏感性增强但对碱产生交叉抗性。冷诱导后的两株菌在氯化钠中的存活率均降低,S.aureus W3最低下降了24.09%。另外,冷诱导后金黄色葡萄球菌对酒精的耐受性增强。分离株S.aureus W3在冷诱导前后过氧化氢耐受性没有明显变化(p>0.05),但标准菌株S.aureus CICC10201敏感性增强,存活率显著下降(p<0.05)。分离株S.aureus W3较S.aureus CICC10201具有更强的生物被膜形成能力,运用1% 84消毒水能有效清除其生物被膜。本研究为冷冻食品中金黄色葡萄球菌的安全控制提供理论依据。     

Mercury reduction and oxidation by reduced natural organic matter in anoxic environments

Natural organic matter (NOM)-mediated redox cycling of elemental mercury Hg(0) and mercuric Hg(II) is critically important in affecting inorganic mercury transformation and bioavailability. However, these processes are not well understood, particularly in anoxic water and sediments where NOM can be reduced and toxic methylmercury is formed. We show that under dark anoxic conditions reduced organic matter (NOM(re)) simultaneously reduces and oxidizes Hg via different reaction mechanisms. Reduction of Hg(II) is primarily caused by reduced quinones. However, Hg(0) oxidation is controlled by thiol functional groups via oxidative complexation, which is demonstrated by the oxidation of Hg(0) by low-molecular-weight thiol compounds, glutathione, and mercaptoacetic acid, under reducing conditions. Depending on the NOM source, oxidation state, and NOM:Hg ratio, NOM reduces Hg(II) at initial rates ranging from 0.4 to 5.5 h(-1), which are about 2 to 6 times higher than those observed for photochemical reduction of Hg(II) in open surface waters. However, rapid reduction of Hg(II) by NOM(re) can be offset by oxidation of Hg(0) with an estimated initial rate as high as 5.4 h(-1). This dual role of NOM(re) is expected to strongly influence the availability of reactive Hg and thus to have important implications for microbial uptake and methylation in anoxic environments.     

Estimation of the major source and sink of methylmercury in the Florida Everglades

Mercury methylation and/or demethylation have been observed in several compartments [soil (saturated soils covered by standing water), floc, periphyton, and water] of the Everglades, a wetland with mercury as one of the major water quality concerns. However, it is still unclear which compartment is the major source or sink due to the lack of estimation and comparison of the net methylmercury (MeHg) production or degradation in these compartments. The lack of this information has limited our understanding of Hg cycling in this ecosystem. This study adopted a double stable isotope ((199)Hg(2+) and Me(201)Hg) addition technique to determine the methylation/demethylation rate constants and the net MeHg production rates in each compartment. This study improved the previous models for estimating these parameters by (1) taking into account the difference between newly input and ambient mercury in methylation/demethylation efficiency and (2) correcting the contribution of photodemethylation to Me(199)Hg concentration when calculating methylation rates in water. The net MeHg production rate in each compartment was then estimated to identify the major sources and sinks of MeHg. The results indicate that these improvements in modeling are necessary, as a significant error would occur otherwise. Soil was identified to be the largest source of MeHg in the Everglades, while the floc and water column were identified as the major sinks. The role of periphyton varies, appearing to be a source in the northern Everglades and a sink in the southern Everglades. Soil could be the largest source for MeHg in the water column, while methylation in periphyton could also contribute significantly in the northern Everglades.     

Effect of chemical speciation on toxicity of mercury to Escherichia coli biofilms and planktonic cells

While it is known that microbial uptake of mercury (Hg) by planktonic cultures is influenced by the extracellular speciation of mercury in aquatic systems, Hg uptake in biofilm cultures is understudied. We compared the importance of Hg(II) speciation in toxicity to both planktonic and biofilm cultures of the Gram-negative bacterium Escherichia coli 055. Variable chloride chemistry experiments were carried out to modify mercury speciation. Biofilms were observed to be more resistant to Hg than planktonic cells. In both planktonic and biofilm cultures, the toxicity of Hg increased and then decreased along the chloride gradient. The percent reduction in cell viability was linearly related to the concentration of HgCl2(0) when Hg-chloro complexes dominated the speciation, consistent with a passive diffusion model. However, toxicity to both planktonic cells and biofilms at low salinities could not be explained by passive diffusion alone, which suggests that microbial uptake of Hg in both planktonic cells and biofilms may occur by both passive diffusion of neutral species and facilitated uptake. The relationship between toxicity and chloride concentration was similar in the presence and absence of a biofilm, indicating that the presence of the biofilm does not drastically change the relative availability of the dominant mercury species.     

Temperature Acclimation and Its Effects on Porcine Muscle Properties in Two Humidity Environments

SUMMARY— The effects of temperature and humidity on postmortem and associated muscle properties during growth of "stress susceptible" pigs were evaluated. Exposure to ambient temperatures of 32 and 21°C for alternating 3-day periods caused rapid post-mortem glycolysis, high percent light reflectance, and increased light to dark fiber ratios in the longissimus dorsi muscle as compared to constant (27°C) temperature, but only in moderate (38–42% relative) humidity environments. The above events due to temperature acclimation were masked when the humidity was low (17–23% relative). Humidity effects that were independent of temperature acclimation resulted in high percent light reflectance and high muscle temperature in the post-mortem muscle of pigs reared in low humidity. No significant differences were found in lactic dehydrogenase or succinic dehydrogenase enzyme activities of longissimus dorsi or gluteus medius muscles.     

Behavior of flavonols and carotenoids of minimally processed kale leaves during storage in passive modified atmosphere packaging

Minimally processed kale leaves were packed in passive modified atmosphere and stored at 3 conditions: 1 °C in the dark and 11 °C with or without light exposure. The products were evaluated during storage in terms of headspace gas composition, sensory attributes, flavonol, and carotenoid contents. The sensory quality decreased slightly during 17 d at 1 °C in the dark. At 11 °C, the vegetable shelf life was predicted to be 6 d in the dark and 3 d with light. Quercetin and kaempferol were stable during storage for 15 d at 1 °C in the absence of light. At 11 °C in the dark, quercetin was stable during 10 d, increasing slightly on the 8th day. Kaempferol decreased up to the 5th day but increased on the 8th day, decreasing again on the 10th day. After 5 d at 11 °C under light, the flavonol levels were significantly higher than those of the initial values. Neoxanthin and violaxanthin did not change significantly after 15 d at 1 °C in the dark. Lutein and β-carotene, however, decreased 7.1% and 11.3%, respectively. At 11 °C in the dark, neoxanthin, violaxanthin, lutein, and β-carotene decreased 16.1%, 13.2%, 24.1%, and 23.7% after 10 d, respectively. At 11 °C under light, neoxanthin and lutein had a slight increase while violaxanthin and β-carotene decreased 23.1% and 16.5% after 5 d. Practical Application: Passive modified atmosphere packaging together with refrigeration can extend the shelf life of minimally processed kale, retaining the health-promoting compounds, flavonols and carotenoids. Quercetin, kaempferol, neoxanthin, and violaxanthin are stable and lutein and β-carotene slightly reduced.     

Organic material: the primary control on mercury methylation and ambient methyl mercury concentrations in estuarine sediments

Estuarine environments that have no direct sources of mercury (Hg) pollution may have sediment concentrations of methylmercury (MeHg) as high as those of polluted marine environments. In this study we examined the biogeochemical factors affecting net methylation and sediment MeHg concentrations in an unpolluted estuarine environment, the Ore River estuary, which discharges into the Bothnian Bay (20-120 ng total Hg g(-1) dry sediment, salinity 3-5% per hundred). We analyzed the spatial and temporal differences in surface sediment profiles of MeHg concentration, Hg methylation, MeHg demethylation, and concentrations of sulfide and oxygen between accumulation and erosion type bottoms. The main difference between the bottoms studied was in the proportion of organic material (OM) in the sediment, ranging between 0.8% and 10.8%. The pore water sulfide concentration profiles also differed considerably between sites and seasons, from 0 to 20 microM, with 100 microM as the extreme maximum. The sediment MeHg concentration profiles (0-10 cm) mostly varied between 0.1 and 7 ng g(-1) dry weight (dw, as Hg). The MeHg demethylation rates were relatively low and the depth profiles of the rates were relatively constant over season, site, and depth. In contrast, both rates and depths of maximum Hg methylation differed between the bottoms. The results indicate that the amount of OM accumulated at the bottoms was the main factor affecting net MeHg production, while the total amount of Hg had little or no influence on the amount of MeHg in the sediment.     

Antioxidant activity of 1,4-dihydropyridine derivatives in β-carotene-methyl linoleate,sunflower oil and emulsions

The antioxidant activity of six synthetic 1,4-dihydropyridine (DHP) derivatives was tested in an azobis-amidinopropane dihydrochloride initiated β-carotene-methyl linoleate peroxidation model system. Radical scavenging activity of the derivatives was estimated by measuring their reactivity with stable N,N-diphenyl-N′-picrylhydrazyl radicals. The antioxidant activity of these different derivatives was also evaluated in traditional sunflower oil and its 20% oil-in-water emulsion in the dark at 60°C, as well as during storage of sunflower oil in the dark and with light exposure at ambient temperatures. The primary (conjugated diene hydroperoxides) and secondary (hexanal, pentanal) oxidation products were monitored during different periods of storage. Butylated hydroxytoluene was used as a standard antioxidant and different plant extracts were used for comparison of antioxidant activities throughout this study. All synthetic compounds showed antioxidant activity, while plant extracts acted as pro-oxidants upon light exposure. Three derivatives of DHP, without substitution at position 4, showed highest antioxidative activity in model system and sunflower oil and its emulsion in the dark at 60°C as well as during storage of sunflower oil in the dark and with light exposure at ambient temperature. The hydrophilic derivative of DHP with a phenyl group at position 4 showed high radical scavenging activity, and high antioxidant activity in sunflower oil-in-water emulsion, but was less active in model system and sunflower oil. The hydrophobic derivatives of DHP with a phenyl group at position 4 showed high antioxidant activity in model system but were the lowest active derivatives in sunflower oil and its emulsion. ©     

冻干甜玉米粒贮藏过程中玉米黄质稳定性分析

冻干甜玉米粒真空包装和普通包装后置于4、20、37 ℃条件下贮藏12 周,考察包装方式和贮藏条件对冻干 甜玉米粒中玉米黄质稳定性的影响。采用高效液相色谱方法测定玉米黄质含量。结果表明：普通包装、高温和见 光均可加速玉米黄质降解,且遵循一级反应动力学模型;不同条件反应速率不同,真空包装4 ℃避光贮藏玉米黄质 降解速率常数最小,普通包装37 ℃见光贮藏降解速率常数最大,且普通包装见光贮藏对玉米黄质热敏感性影响最 大。贮藏过程中玉米黄质会发生顺反异构化,真空包装避光和见光时温度越高生成的顺式异构体越多;同一温度条 件下,真空包装见光比避光更易发生反式向顺式结构的转变;普通包装避光或见光贮藏时玉米黄质以降解反应为 主,异构化反应不明显。     

Effect of loading rate on the fate of mercury in littoral mesocosms

The effects of changes in atmospheric mercury (Hg) deposition on aquatic ecosystems are poorly understood. In this study, we examined the biogeochemical cycling of Hg in littoral mesocosms receiving different loading rates (7-107 microg Hg m(-2) year(-1)). We added a 202Hg-enriched preparation to differentiate the experimentally added Hg from the ambient Hg in the environment. This approach allowed us to follow the distribution and methylation of the isotopically enriched ("spike") Hg in the mesocosms. Within 3 weeks, spike Hg was distributed throughout the main environmental compartments (water, particles, periphyton, and sediments) and began to be converted to methylmercury (MeHg). Concentrations of spike total Hg and MeHg in these compartments, measured after 8 weeks, were directly proportional to loading rates. Thus, Hg(II) availability was the limiting factor for the major processes of the biogeochemical Hg cycle, including methylation. This is the first study to demonstrate a proportional response of in situ MeHg production to atmospherically relevant loading levels. On the basis of mass balances, we conclude that loading rate had no effect on the relative distribution of spike Hg among the main compartments or on the fraction of spike Hg converted to MeHg. Therefore, loading rate did not change the relative magnitude of biogeochemical pathways competing for Hg within the mesocosms. These data suggest that reductions of Hg deposition to lake surfaces would be equally effective across a broad range of deposition rates.     

Decrease in net mercury methylation rates following iron amendment to anoxic wetland sediment slurries

The rate of mercury methylation in anoxic wetland sediments is affected by the concentration of bioavailable complexes between Hg and sulfide. Previous research with pure bacterial cultures has shown that addition of ferrous iron reduces the net rate of mercury methylation by decreasing the concentration of dissolved sulfide. To assess the possibility of using this approach to decrease net mercury methylation in restored and constructed wetlands, laboratory experiments were conducted by adding Hg(II) and Fe(II) to sediments collected from six sites in five estuarine wetlands. Addition of 30 mM (0.07 mmol g(-1) or 3.9 mg g(-1)) Fe(II) decreased net mercury methylation relative to that of unamended controls by a factor of 2.1-6.6. In all cases, the observed decrease in net mercury methylation was accompanied by a decrease in the concentrations of sulfide and filterable mercury in the water overlying the sediments. When iron was added to one of the sediment samples at doses that were small relative to the concentration of sulfide present, net mercury methylation either increased slightly or was unaffected. Comparison of the results to speciation model predictions suggests that dissolved reduced sulfur-containing species play a role in the formation of uncharged, bioavailable Hg complexes. Although further research is needed to determine the long-term effect of iron amendment, these results suggest that iron addition decreases mercury methylation in authentic wetland sediments.