Chemical and microbiological studies of sulfide‐mediated manganese reduction 1

Laboratory studies of manganese reduction by naturally occurring reduced inorganic compounds were undertaken, both to study further possible in situ mechanisms of manganese reduction and to examine how manganese redox reactions might be coupled to other biogeochemical processes. Chemical manganese reduction by sulfide (in the presence of excess manganese oxide) was found to be rapid and complete, with all sulfide being oxidized within 5–10 min. The reduction of δMnO₂ by sulfide involves a two‐electron transfer, with S° the predominant oxidized sulfur product. Using a marine sulfate‐reducing bacterium (Desulfovibrio sp.), the kinetics of sulfide‐dependent, bacterially mediated manganese reduction were studied; the rate‐limiting step was bacterial sulfide production. These findings suggest that in stratified marine environments (such as the Black Sea, Saanich Inlet, or certain coastal sediments) manganese reduction should occur just below the oxic‐anoxic (O₂/H₂S) interface or redox boundary as a result of the chemical reaction between manganese oxides and sulfide produced by sulfate‐reducing bacteria.     

Dominance of sulfur-fueled iron oxide reduction in low-sulfate freshwater sediments

A central tenant in microbial biogeochemistry is that microbial metabolisms follow a predictable sequence of terminal electron acceptors based on the energetic yield for the reaction. It is thereby oftentimes assumed that microbial respiration of ferric iron outcompetes sulfate in all but high-sulfate systems, and thus sulfide has little influence on freshwater or terrestrial iron cycling. Observations of sulfate reduction in low-sulfate environments have been attributed to the presumed presence of highly crystalline iron oxides allowing sulfate reduction to be more energetically favored. Here we identified the iron-reducing processes under low-sulfate conditions within columns containing freshwater sediments amended with structurally diverse iron oxides and fermentation products that fuel anaerobic respiration. We show that despite low sulfate concentrations and regardless of iron oxide substrate (ferrihydrite, Al-ferrihydrite, goethite, hematite), sulfidization was a dominant pathway in iron reduction. This process was mediated by (re)cycling of sulfur upon reaction of sulfide and iron oxides to support continued sulfur-based respiration—a cryptic sulfur cycle involving generation and consumption of sulfur intermediates. Although canonical iron respiration was not observed in the sediments amended with the more crystalline iron oxides, iron respiration did become dominant in the presence of ferrihydrite once sulfate was consumed. Thus, despite more favorable energetics, ferrihydrite reduction did not precede sulfate reduction and instead an inverse redox zonation was observed. These findings indicate that sulfur (re)cycling is a dominant force in iron cycling even in low-sulfate systems and in a manner difficult to predict using the classical thermodynamic ladder.     

Biogeochemistry of an Iron-Rich Hypersaline Microbial Mat (Camargue, France)

In situ microsensor measurements were combined with biogeochemical methods to determine oxygen, sulfur, and carbon cycling in microbial mats growing in a solar saltern (Salin-de-Giraud, France). Sulfate reduction rates closely followed the daily temperature changes and were highest during the day at 25°C and lowest during the night at 11°C, most probably fueled by direct substrate interactions between cyanobacteria and sulfate-reducing bacteria. Sulfate reduction was the major mineralization process during the night and the contribution of aerobic respiration to nighttime DIC production decreased. This decrease of aerobic respiration led to an increasing contribution of sulfide (and iron) oxidation to nighttime O₂ consumption. A peak of elemental sulfur in a layer of high sulfate reduction at low sulfide concentration underneath the oxic zone indicated anoxygenic photosynthesis and/or sulfide oxidation by iron, which strongly contributed to sulfide consumption. We found a significant internal carbon cycling in the mat, and sulfate reduction directly supplied DIC for photosynthesis. The mats were characterized by a high iron content of 56 mol Fe cm^–3, and iron cycling strongly controlled the sulfur cycle in the mat. This included sulfide precipitation resulting in high FeS contents with depth, and reactions of iron oxides with sulfide, especially after sunset, leading to a pronounced gap between oxygen and sulfide gradients and an unusual persistence of a pH peak in the uppermost mat layer until midnight.     

Effect of sulfate on carbon and electron flow during microbial methanogenesis in freshwater sediments.

The effect of sulfate on methane production in Lake Mendota sediments was investigated to clarify the mechanism of sulfate inhibition of methanogenesis. Methanogenesis was shown to be inhibited by the addition of as little as 0.2 mM sulfate. Sulfate inhibition was reversed by the addition of either H2 or acetate. Methane evolved when inhibition was reversed by H2 additions was derived from 14CO2. Conversely, when acetate was added to overcome sulfate inhibition, the evolved methane was derived from [2-14C]acetate. A competition for available H2 and acetate was proposed as the mechanism by which sulfate inhibited methanogenesis. Acetate was shown to be metabolized even in the absence of methanogenic activity. In the presence of sulfate, the methyl position of acetate was converted to CO2. The addition of sulfate to sediments did not result in the accumulation of significant amounts of sulfide in the pore water. Sulfate additions did not inhibit methanogenesis unless greater than 100 mug of free sulfide per ml was present in the pore water. These results indicate that carbon and electron flow are altered when sulfate is added to sediments. Sulfate-reducing organisms appear to assume the role of methanogenic bacteria in sulfate-containing sediments by utilizing methanogenic precursors.     

Metal-dependent anaerobic methane oxidation in marine sediment: Insights from marine settings and other systems

Anaerobic oxidation of methane(AOM) plays a crucial role in controlling global methane emission. This is a microbial process that relies on the reduction of external electron acceptors such as sulfate, nitrate/nitrite, and transient metal ions. In marine settings, the dominant electron acceptor for AOM is sulfate, while other known electron acceptors are transient metal ions such as iron and manganese oxides. Despite the AOM process coupled with sulfate reduction being relatively well characterized,researches on metal-dependent AOM process are few, and no microorganism has to date been identified as being responsible for this reaction in natural marine environments. In this review, geochemical evidences of metal-dependent AOM from sediment cores in various marine environments are summarized. Studies have showed that iron and manganese are reduced in accordance with methane oxidation in seeps or diffusive profiles below the methanogenesis zone. The potential biochemical basis and mechanisms for metal-dependent AOM processes are here presented and discussed. Future research will shed light on the microbes involved in this process and also on the molecular basis of the electron transfer between these microbes and metals in natural marine environments.     

Bacterial Disproportionation of Elemental Sulfur Coupled to Chemical Reduction of Iron or Manganese

A new chemolithotrophic bacterial metabolism was discovered in anaerobic marine enrichment cultures. Cultures in defined medium with elemental sulfur (S⁰) and amorphous ferric hydroxide (FeOOH) as sole substrates showed intense formation of sulfate. Furthermore, precipitation of ferrous sulfide and pyrite was observed. The transformations were accompanied by growth of slightly curved, rod-shaped bacteria. The quantification of the products revealed that S⁰ was microbially disproportionated to sulfate and sulfide, as follows: 4S⁰ + 4H₂O → SO₄^2- + 3H₂S + 2H⁺. Subsequent chemical reactions between the formed sulfide and the added FeOOH led to the observed precipitation of iron sulfides. Sulfate and iron sulfides were also produced when FeOOH was replaced by FeCO₃. Further enrichment with manganese oxide, MnO₂, instead of FeOOH yielded stable cultures which formed sulfate during concomitant reduction of MnO₂ to Mn²⁺. Growth of small rod-shaped bacteria was observed. When incubated without MnO₂, the culture did not grow but produced small amounts of SO₄^2- and H₂S at a ratio of 1:3, indicating again a disproportionation of S⁰. The observed microbial disproportionation of S⁰ only proceeds significantly in the presence of sulfide-scavenging agents such as iron and manganese compounds. The population density of bacteria capable of S⁰ disproportionation in the presence of FeOOH or MnO₂ was high, > 10⁴ cm^-3 in coastal sediments. The metabolism offers an explanation for recent observations of anaerobic sulfide oxidation to sulfate in anoxic sediments.     

Characterization of Enrichment Cultures Involved in the Carbon,Sulfur and Iron Biogeochemical Cycles in French Guiana Mobile Muds

The fluidized sediment ecosystem off French Guiana is characterized by active physical reworking, diversity of electron acceptors and highly variable redox regime. It is well studied geochemically but little is known about specific microorganisms involved in its biogeochemistry. Based on the biogeochemical profiles and rate kinetics, several possible biotically mediated pathways of the carbon, sulfur and iron cycles were hypothesized. Enrichment studies were set up with a goal to culture microorganisms responsible for these pathways. Stable microbial consortia potentially capable of the following chemolithoautotrophic types were enriched from the environment and characterized: elemental sulfur/thiosulfate disproportionators, thiosulfate-oxidizing ferrihydrite and nitrate reducers, sulfide/ferrous sulfide oxidizers coupled with nitrate and microaerophilic iron oxidizers. Attempts to generate several enrichments (anoxic ammonia oxidation, and sulfide oxidizers with ferric iron or manganese oxide) were not successful. Heterotrophic sulfate and elemental sulfur reduction bacteria are prominent and dominate reductive sulfur transformations. We hypothesize that carbon dioxide fixation coupled with synthesis of organic matter happens mostly via sulfur disproportionation and sulfur species oxidation with iron oxidation playing a minor role.     

Seasonal oscillation of microbial iron and sulfate reduction in saltmarsh sediments (Sapelo Island, GA, USA)

Seasonal variations in anaerobic respiration pathways were investigated at three saltmarsh sites using chemical data, sulfate reduction rate measurements, enumerations of culturable populations of anaerobic iron-reducing bacteria (FeRB), and quantification of in situ 16S rRNA hybridization signals targeted for sulfate-reducing bacteria (SRB). Bacterial sulfate reduction in the sediments followed seasonal changes in temperature and primary production of the saltmarsh, with activity levels lowest in winter and highest in summer. In contrast, a dramatic decrease in the FeRB population size was observed during summer at all sites. The collapse of FeRB populations during summer was ascribed to high rates of sulfide production by SRB, resulting in abiotic reduction of bioavailable Fe(III) (hydr)oxides. To test this hypothesis, sediment slurry incubations at 10, 20 and 30 °C were carried out. Increases in temperature and labile organic carbon availability (acetate or lactate additions) increased rates of sulfate reduction while decreasing the abundance of culturable anaerobic FeRB. These trends were not reversed by the addition of amorphous Fe(III) (hydr)oxides to the slurries. However, when sulfate reduction was inhibited by molybdate, no decline in FeRB growth was observed with increasing temperature. Addition of dissolved sulfide adversely impacted propagation of FeRB whether molybdate was added or not. Both field and laboratory data therefore support a sulfide-mediated limitation of microbial iron respiration by SRB. When total sediment respiration rates reach their highest levels during summer, SRB force a decline in the FeRB populations. As sulfate reduction activity slows down after the summer, the FeRB are able to recover.     

Pro- or anti-oxidant manganese: a suggested mechanism for reconciliation

The neurodegeneration induced by manganese has been attributed to its ability to undergo redox cycling, and catalysis of reactive oxygen species (ROS) formation, as with other transition metals. However, the characterization of manganese as a pro-oxidant is confounded by increasing evidence that the metal may scavenge superoxide anions and protect cells from oxidative damage. The current study was designed to address conflicting reports pertaining to the oxidative capacity of manganese. We found that the metal has distinctive redox dynamics in which the divalent reduced form, unlike iron, possessed no intrinsic oxidative capacity. The apparent ability of Mn(2+) to promote the formation of ROS within a cortical mitochondrial-synaptosomal fraction was quenched by the depletion of contaminating nanomolar concentrations of trivalent metals. The addition of manganic ions at trace concentrations dose-dependently restored the oxidative capacity attributed to divalent manganese, whereas the presence of the ferric ion retarded the rate of ROS generation. This result was paralleled by the spectrophotometric demonstration that the kinetics of iron oxidation is accelerated by trivalent but not divalent manganese. The markedly different capacities of the lower and higher valence states of manganese to promote free-radical formation in cortical fractions and to modulate the process of iron oxidation may account for earlier contradictory reports of anti- and pro-oxidant properties of manganese.     

Influences of the petroleum-recovering activity on the arsenic level in groundwater of Kuitun,Xinjiang, China

Increasing demands of groundwater in petroleum-recovering regions could elevate the level and mobility of arsenic in groundwater as a result of the enhanced dissolution of arsenic-bearing iron or manganese oxide due to the accelerated sulfate reduction by microorganisms in a reductive environment. To substantiate this possibility, groundwater samples were collected from 220 water wells in the nearby petroleum wells in Kuitun. Dissolved arsenic, iron, manganese, and sulfate levels and pH in groundwater samples were analyzed. The dissolved arsenic levels in groundwater varied from <2.3 to 789.4 μg·L^?1, in which approximately 96.4% of the measured values exceeded the allowed limits of the World Health Organization. An inverse relation existed between dissolved arsenic and sulfate levels. Most of the high arsenic-level samples (>300 μg·L^?1) were found in wells at close proximity to petroleum wells where a high iron or manganese level was also detected. The oil-exploring activity in the study region seemed to have enhanced the microbial reduction of sulfate in underground environment and hence the level of arsenic in groundwater. The microbial sulfate reduction coupled with the reduction of arsenic-bearing iron oxides in the groundwater environment may explain the spatial heterogeneity of the arsenic level in groundwater.     

Increased removal of nickel and organics in the upflow anaerobic sludge bed filter reactor using sulfate and sulfide additions

Glucose (total organic carbon: 1,200 mg/l) and nickel (0 to 40 mg/l) were added to an anaerobic upflow sludge bed filter reactor. The removal efficiencies of total organic carbon and nickel were maintained to 95% and 98.5%, respectively, since nickel was precipitated with sulfide which was converted from sulfate added at 80 mg SO4-S/l by sulfate reducing bacteria. Sulfate therefore enhances its organic removal efficiency of AUBF reactor under the presence of heavy metal.     

Uptake of six heavy metals by oat as influenced by soil type and additions of cadmium,lead, zinc and copper

Summary The influence of heavy metal additions on availability and uptake of cadmium, lead, zinc, copper, manganese and iron by oat was studied. The experiments were carried out as pot experiments using sandy loam, sandy soil and organic soil. Selective extractants were used to remove metals held in different soil fractions.Lead and copper were preferently bound by organics and oxides, zinc by oxides and inorganics, and cadmium by inorganics and organics.Addition of cadmium to the soils resulted in higher cadmium concentrations in all plant parts but lower concentrations of lead, zinc, copper, manganese and iron, and the accumulation indexes of these metals were also lower when cadmium was added to the soil.Addition of cadmium plus lead, zinc and copper resulted in higher cadmium concentrations in leaves and straw of plants grown in sandy loam and sandy soil, but lower concentrations when plants were grown in organic soil as compared with the results when cadmium was added separately. The transfer of cadmium, lead, zinc and copper from soil to plant was greatest from sandy soil, and zinc and cadmium were more mobile in the plant than were lead and copper.Cadmium concentrations in leaves correlated significantly with CaCl₂ and CH₃COOH extractions in sandy loam and sandy soil and with CH₃COOH extractions in organic soil.Generally, the total metal uptake was lowest from organic soil.     

Microbial Sulfate Reduction and Its Potential Utility as an Acid Mine Water Pollution Abatement Procedure

The presence of high concentrations of sulfate, iron, and hydrogen (acid) ions in drainage from coal mines and other areas containing waste pyritic materials is a serious water pollution problem. Sulfate can be removed from solution by microbial reduction to sulfide and subsequent precipitation as FeS. A mixed culture of microorganisms degraded wood dust cellulose, and the degradation products served as carbon and energy sources for sulfate-reducing bacteria. Metabolism of carbon compounds resulted in a net pH increase in the system. Oxidation-reduction potential (Eh) and temperature and carbon supplements were studied in an effort to accelerate the sulfate reduction process, with the ultimate objective of utilizing the process as a pollution abatement procedure.     

Advances in biotreatment of acid mine drainage and biorecovery of metals: 1. Metal precipitation for recovery and recycle

Acid mine drainage (AMD), an acidic metal-bearingwastewater, poses a severe pollution problem attributedto post mining activities. The metals usuallyencountered in AMD and considered of concern for riskassessment are arsenic, cadmium, iron, lead, manganese,zinc, copper and sulfate. The pollution generated byabandoned mining activities in the area of Butte, Montanahas resulted in the designation of the Silver Bow Creek–ButteArea as the largest Superfund (National Priorities List) sitein the U.S. This paper reports the results of bench-scalestudies conducted to develop a resource recovery basedremediation process for the clean up of the Berkeley Pit.The process utilizes selective, sequential precipitation (SSP)of metals as hydroxides and sulfides, such as copper, zinc,aluminum, iron and manganese, from the Berkeley Pit AMDfor their removal from the water in a form suitable foradditional processing into marketable precipitates and pigments.The metal biorecovery and recycle process is based on completeseparation of the biological sulfate reduction step and themetal precipitation step. Hydrogen sulfide produced in the SRBbioreactor systems is used in the precipitation step to forminsoluble metal sulfides. The average metal recoveries usingthe SSP process were as follows: aluminum (as hydroxide) 99.8%,cadmium (as sulfide) 99.7%, cobalt (as sulfide) 99.1% copper(as sulfide) 99.8%, ferrous iron (sulfide) 97.1%, manganese(as sulfide) 87.4%, nickel (as sulfide) 47.8%, and zinc (as sulfide)100%. The average precipitate purity for metals, copper sulfide,ferric hydroxide, zinc sulfide, aluminum hydroxide and manganesesulfide were: 92.4, 81.5, 97.8, 95.6 , 92.1 and 75.0%, respectively.The final produced water contained only calcium and magnesiumand both sulfate and sulfide concentrations were below usablewater limits. Water quality of this agriculturally usable watermet the EPA's gold standard criterion.     

Acetate as the main energy substrate for the sulfate-reducing bacteria in Lake Eliza (South Australia) hypersaline sediments

Abstract Simultaneous measurements of sulfate reduction and acetate oxidation using ³⁵S and ¹⁴C tracers showed that acetate was the main energy substrate for the sulfate-reducing bacteria in Lake Eliza sediments. Sulfate reduction rates calculated from acid-volatile sulfide data only, correlated with acetate oxidation at around 0.5:1. However, the rates calculated from acid-volatile plus pyrite sulfur data correlated with acetate oxidation at a ratio of around 1:1. Molybdate completely inhibited sulfate reduction but acetate oxidation was not totally inhibited. From 10 to 15% of acetate oxidation was not attributable to the sulfate-reducing bacteria. There was rapid accumulation of acetate, within the first 12 h of incubation. Acetate, propionate and butyrate accumulated in the presence of molybdate.     

Terminal Processes in the Anaerobic Degradation of an Algal-Bacterial Mat in a High-Sulfate Hot Spring

The algal-bacterial mat of a high-sulfate hot spring (Bath Lake) provided an environment in which to compare terminal processes involved in anaerobic decomposition. Sulfate reduction was found to dominate methane production, as indicated by comparison of initial electron flow through the two processes, rapid conversion of [2-¹⁴C]acetate to ¹⁴CO₂ and not to ¹⁴CH₄, and the lack of rapid reduction of NaH¹⁴CO₃ to ¹⁴CH₄. Sulfate reduction was the dominant process at all depth intervals, but a marked decrease of sulfate reduction and sulfate-reducing bacteria was observed with depth. Concurrent methanogenesis was indicated by the presence of viable methanogenic bacteria and very low but detectable rates of methane production. A marked increased in methane production was observed after sulfate depletion despite high concentrations of sulfide (>1.25 mM), indicating that methanogenesis was not inhibited by sulfide in the natural environment. Although a sulfate minimum and sulfide maximum occurred in the region of maximal sulfate reduction, the absence of sulfate depletion in interstitial water suggests that methanogenesis is always severely limited in Bath Lake sediments. Low initial methanogenesis was not due to anaerobic methane oxidation.     

Arsenic Bioremediation by Biogenic Iron Oxides and Sulfides

Microcosms containing sediment from an aquifer in Cambodia with naturally elevated levels of arsenic in the associated groundwater were used to evaluate the effectiveness of microbially mediated production of iron minerals for in situ As remediation. The microcosms were first incubated without amendments for 28 days, and the release of As and other geogenic chemicals from the sediments into the aqueous phase was monitored. Nitrate or a mixture of sulfate and lactate was then added to stimulate biological Fe(II) oxidation or sulfate reduction, respectively. Without treatment, soluble As concentrations reached 3.9 ± 0.9 μM at the end of the 143-day experiment. However, in the nitrate- and sulfate-plus-lactate-amended microcosms, soluble As levels decreased to 0.01 and 0.41 ± 0.13 μM, respectively, by the end of the experiment. Analyses using a range of biogeochemical and mineralogical tools indicated that sorption onto freshly formed hydrous ferric oxide (HFO) and iron sulfide mineral phases are the likely mechanisms for As removal in the respective treatments. Incorporation of the experimental results into a one-dimensional transport-reaction model suggests that, under conditions representative of the Cambodian aquifer, the in situ precipitation of HFO would be effective in bringing groundwater into compliance with the World Health Organization (WHO) provisional guideline value for As (10 ppb or 0.13 μM), although soluble Mn release accompanying microbial Fe(II) oxidation presents a potential health concern. In contrast, production of biogenic iron sulfide minerals would not remediate the groundwater As concentration below the recommended WHO limit.     

Characteristics of assimilatory sulfate transport in Rhodobacter sulfidophilus

Abstract Sulfate uptake was investigated with four species of phototrophic sulfur bacteria. Rhodobacter sulfidophilus and Chromatium vinosum took up ³⁵S-labeled sulfate added in micromolar concentrations. Sulfate uptake by C. vinosum was expressed only under sulfate starvation. R. sulfidophilus took up 10 μM sulfate almost completely and accumulated it up to 5300-fold, also when grown with excess sulfate. Sulfite (1 mM) as an intermediate of sulfate assimilation inhibited sulfate uptake completely within 1 min. Moderate inhibition was observed with cysteine (1 mM) and none with sulfide (1 mM). Transport was not dependent on the cations K⁺, Na⁺, Li⁺ or protons, but was sensitive to uncouplers and to the ATPase inhibitor dicyclohexylcarbodiimide (DCCD). The accumulation of sulfate correlated with the ATP concentration in the cells, indicating an ATP-dependent uptake mechanism.     

Manganese and iron oxidation by fungi isolated from building stone

Acid and nonacid generating fungal strains isolated from weathered sandstone, limestone, and granite of Spanish cathedrals were assayed for their ability to oxidize iron and manganese. In general, the concentration of the different cations present in the mineral salt media directly affected Mn(IV) oxide formation, although in some cases, the addition of glucose and nitrate to the culture media was necessary. Mn(II) oxidation in acidogenic strains was greater in a medium containing the highest concentrations of glucose, nitrate, and manganese. High concentrations of Fe(II), glucose, and mineral salts were optimal for iron oxidation. Mn(IV) precipitated as oxides or hydroxides adhered to the mycelium. Most of the Fe(III) remained in solution by chelation with organic acids excreted by acidogenic strains. Other metabolites acted as Fe(III) chelators in nonacidogenic strains, although Fe(III) deposits around the mycelium were also detected. Both iron and manganese oxidation were shown to involve extracellular, hydrosoluble enzymes, with maximum specific activities during exponential growth. Strains able to oxidize manganese were also able to oxidize iron. It is concluded that iron and manganese oxidation reported in this work were biologically induced by filamentous fungi mainly by direct (enzymatic) mechanisms.Correspondence to: G. Gomez-Alarcon.     

2,4-二氯苯酚的矿物表面化学行为研究

矿物质是土壤中的重要组成成分,本实验采用动态反应和平衡反应研究氧化物矿物表面2,4-二氯苯酚(2,4-DCP)的化学行为.结果表明,无定形铁氧化物、针铁矿、α-MnO₂、δ-MnO₂对自然的或外源的有机化合物有较强的氧化能力,其中对2,4二氯苯酚的氧化能力是:δ-MnO₂αMnO₂>无定形铁氧化物、针铁矿.采用分段法估算反应系统的速率,发现氧化锰氧化2,4-二氯苯酚的反应速率是氧化铁的3～50倍,δ-MnO₂氧化2,4-二氯苯酚的速率是α-MnO₂的1.5～3.2倍.根系分泌物由于参与氧化物表面化学反应,对系统2,4-二氯苯酚的氧化过程产生一定影响,α-MnO₂加入根系分泌物后,2,4-二氯苯酚在溶液中的消减速率明显减慢,δ-MnO₂则相反,在根系分泌物共存下溶液中的2,4-二氯苯酚的转化率增加.Cu复合共存可能影响2,4二氯苯酚在矿物表面的键合氧化过程,其中δ-MnO₂对铜吸附能力大于αMnO₂,Cu复合共存对δMnO₂表面2,4-二氯苯酚的氧化影响较大.