Treatment of metal-contaminated water using bacterial sulfate reduction: results from pilot-scale reactors

Simple anaerobic reactors were installed to treat metal-contaminated water in an underground coal mine and at a smelting residues dump in Pennsylvania. The reactors consisted of barrels and tanks filled with spent mushroom compost, within which bacterial sulfate reduction became established. Concentrations of Al, Cd, Fe, Mn, Ni, and Zn were typically lowered by over 95% as contaminated water flowed through the reactors. Cadmium, Fe, Ni, and some Zn were retained as insoluble metal sulfides following their reaction with bacterially generated H(2)S. Aluminum, Mn, and some Zn hydrolyzed and were retained as insoluble hydroxides or carbonates. Reactor effluents were typically circumneutral in pH and contained net alkalinity. The principal sources of alkalinity in the reactors were bacterial sulfate reduction and limestone dissolution. This article examines the chemistry of the reactor systems and the opportunities for enhancing their metal-retaining and alkalinity-generating potential. (c) 1992 John Wiley & Sons, Inc.     

Lead removal through biological sulfate reduction process

The feasibility of lead removal through biological sulfate reduction process with ethanol as electron donor was investigated. Sulfide-rich effluent from biological process was used to remove lead as lead sulfide precipitate. The experiments were divided into two stages; Stage I startup and operation of sulfidogenic process in a UASB reactor and Stage II lead sulfide precipitation. In Stage I, the COD:S ratio was gradually reduced from 15:1 to 2:1. At the COD:S ratio of 2:1, sulfidogenic condition was achieved as identified by 80-85% of electron flow by sulfate reducing bacteria (SRB). COD and sulfate removal efficiency were approximately 78% and 50%, respectively. In Stage II, the effluent from UASB reactor containing sulfide in the range of 30-50 mg/L and lead-containing solution of 45-50 mg/L were fed continuously into the precipitation chamber in which the optimum pH for lead sulfide precipitation of 7.5-8.5 was maintained. It was found that lead removal of 85-95% was attained.     

A kinetic study on bacterial sulfate reduction

The activity of anaerobic sulfate reduction was studied using sulfate-reducing bacteria isolated from the water produced from a Brazilian oil reservoir. The effects of the initial sulfate concentration on the anaerobic sulfate reduction and sulfide generation kinetics were investigated. The redox potential, the biomass solution content, and the sulfate and the sulfide solution content were measured. The results indicate that the sulfate conversion and the sulfide generation are both first-order processes for the initial sulfate concentration of 823, 1,282, and 1,790 mg/L. The results for the kinetic constants for the sulfate conversion indicate an inhibition with the enhancement of the initial sulfate solution content. The kinetic constants for the sulfide generation indicate that this reaction is almost independent of the initial sulfate solution content due to the presence of at least two in-series processes that are faster than the microbial conversion of the sulfate. The kinetic test using the water from an onshore oil field, with an initial sulfide content of 228 mg/L and sulfate content of 947 mg/L, shows a sulfate conversion of 50 % in 528 h. The kinetic modeling for the net content of sulfate and sulfide indicates that the sulfate conversion is slower for this water than for the deionized water tests; however, the sulfide formation has almost the same conversion velocity. The reactions are first order in both cases.     

Enhancement of sulfate reduction activity using granular sludge in anaerobic treatment of acid mine drainage

Three laboratory-scale, upflow anaerobic reactors were operated for about 250 d to determine the effect of activated granular sludge with high density of sulfate reducing bacteria in the treatment of artificial acid mine drainage. Sulfate reducing bacteria in the granular sludge taken from the upflow anaerobic sludge blanket reactor were 1–2×10⁶ c.f.u. g^–1, which is at least 10 times higher than that of organic substrates such as cow manure and oak compost. The reactors with granular sludge effectively removed over 99% of heavy metals, such as Fe, Al, Cu, and Cd during the experiment. This result suggests a feasibility of the application of granular sludge as a source of sulfate reducing bacteria for the treatment of acid mine drainage.     

The removal of bacterial biofilm from water-filled tubes using axially propagated ultrasound

Axially propagated ultrasound (APU) has been used to remove mineralized Proteus mirabilis biofilms from water-filled glass tubes. Pulses (30 s) of ultrasound at frequencies ranging from 350 to 20 kHz were applied to one end of the tubes and the extent of biofilm removal was assessed by infra-red absorptiometry. The removal of biofilm from 7 cm tubes by two 30 s pulses of APU at 150 kHz (54·8%) was comparable to that (60·9%) achieved by sonication at 33 kHz in a conventional sonic cleaning bath. In 50 cm tubes, three 30 s pulses of APU from the 20 kHz transducer removed 87·5% of the biofilm. The equivalent results for APU at 150 kHz and 350 kHz were 66·8% and 31·3%, respectively. These observations suggest that the axial propagation of ultrasound along lengths of water-filled tubing offers the prospect of on-line cleaning of systems such as heat-exchanger pipework.     

Methanogenesis and sulfate reduction in timber and drainage water from a gold mine

Biogenesis of methane in the heartwood of diseased trees has been shown, but never in timber in service. Studies were undertaken to establish whether methan‐ogens and sulfate‐reducers were present in wooden pit props and drainage water from underground sites in a gold mine. The predominant methanogen in the mine ecosystem was tentatively identified as Methanobacterium bryantii. The sulfate‐reducers comprised Desulfovibrio desulfuricans and Desulfotomaculum antarcticum. Most probable numbers (MPN) of bacteria indicated that 3.5 × 10⁵ methanogenic and 7.9 × 10³ sulfate‐reducing bacteria were present per milliliter of stagnant drainage water. MPN values per gram of timber were lower for methanogens but comparable for sulfate‐reducers. Laboratory model systems predicted a maximum rate of methan‐ogenesis of 2.3 mL methane/g wood per day; however, rates would never attain this value because of nutrient limitations and environmental restrictions. Analysis of gas samples extracted from sealed areas of the gold mine verified the presence of methane.     

Automated screening of inhibitors of bacterial dissimilatory sulfate reduction

Summary An automated assay system was used to screen inhibitors of sulfide formation by sulfate-reducing bacteria. The system used a Technicon AutoAnalyzer II instrument both as an incubation chamber (to combine bacterial cells, substrates, and test compounds) and as an analyzer of the amount of sulfide formed during incubation. A lactate-limited chemostat was used to provide a reproducible inoculum of Desulfovibrio desulfuricans cells. The assay was useful in detecting even small changes in the rate of sulfate reduction resulting from exposure to inhibitors.Contribution no. 5652 from the Central Research and Development Department Offprint requests to: P. J. Weimer     

Selective removal of transition metals from acidic mine waters by novel consortia of acidophilic sulfidogenic bacteria

Two continuous‐flow bench‐scale bioreactor systems populated by mixed communities of acidophilic sulfate‐reducing bacteria were constructed and tested for their abilities to promote the selective precipitation of transition metals (as sulfides) present in synthetic mine waters, using glycerol as electron donor. The objective with the first system (selective precipitation of copper from acidic mine water containing a variety of soluble metals) was achieved by maintaining a bioreactor pH of ～2.2–2.5. The second system was fed with acidic (pH 2.5) synthetic mine water containing 3 mM of both zinc and ferrous iron, and varying concentrations (0.5–30 mM) of aluminium. Selective precipitation of zinc sulfide was possible by operating the bioreactor at pH 4.0 and supplementing the synthetic mine water with 4 mM glycerol. Analysis of the microbial populations in the bioreactors showed that they changed with varying operational parameters, and novel acidophilic bacteria (including one sulfidogen) were isolated from the bioreactors. The acidophilic sulfidogenic bioreactors provided ‘proof of principle’ that segregation of metals present in mine waters is possible using simple online systems within which controlled pH conditions are maintained. The modular units are versatile and robust, and involve minimum engineering complexity.     

The bacterial communities of bioelectrochemical systems associated with the sulfate removal under different pHs

Sulfate contamination in ecosystems has been a serious problem. Among various technologies, bioelectrochemical systems (BESs) show the advantage of no-pollution and low-cost for removing sulfate. In order to further expound the biological process of sulfate removal in BESs, 454 pyrosequencing was applied to analyze the bacterial communities under different pH conditions. The bacterial community profiles were analyzed from three aspects: (a) the α-diversity and β-diversity of bacterial communities, (b) the distribution of bacterial phylotypes, and (c) the characterizations of dominant operational taxonomic units (OTUs). We demonstrated that the indexes of phylotype richness and phylogenetic diversity were positively correlated across the pH gradient in the BESs. Among the dominant OTUs, the OTUs which were highly similar to Desulfatirhabdium butyrativorans, Desulfovibrio marrakechensis and Desulfomicrobium sp. might participate in removing sulfate. Standing on genus level, Desulfomicrobium and Sulfuricurvum play conducing and adverse roles for sulfate removal in alkaline condition, respectively. Desulfovibrio contributed to removing sulfate in the neutral and acidic conditions, while Thiomonas mainly weakened the performance of sulfate removal in neutral pH condition. These results further clarified how pH condition directly affected the bacterial communities, which consequently affected the performance of sulfate pollutant treatment using BESs.     

Archaeal and bacterial communities of heavy metal contaminated acidic waters from zinc mine residues in Sepetiba Bay

Mining of metallic sulfide ore produces acidic water with high metal concentrations that have harmful consequences for aquatic life. To understand the composition and structure of microbial communities in acid mine drainage (AMD) waters associated with Zn mine tailings, molecular diversity of 16S genes was examined using a PCR, cloning, and sequencing approach. A total of 78 operational taxonomic units (OTUs) were obtained from samples collected at five different sites in and around mining residues in Sepetiba Bay, Brazil. We analyzed metal concentration, physical, chemical, and microbiological parameters related to prokaryotic diversity in low metal impacted compared to highly polluted environments with Zn at level of gram per liter and Cd–Pb at level of microgram per liter. Application of molecular methods for community structure analyses showed that Archaea and Bacteria groups present a phylogenetic relationship with uncultured environmental organisms. Phylogenetic analysis revealed that bacteria present at the five sites fell into seven known divisions, α-Proteobacteria (13.4%), β-Proteobacteria (16.3%), γ-Proteobacteria (4.3%), Sphingobacteriales (4.3%), Actinobacteria (3.2%) Acidobacteria (2.1%), Cyanobacteria (11.9%), and unclassified bacteria (44.5%). Almost all archaeal clones were related to uncultivated Crenarchaeota species, which were shared between high impacted and low impacted waters. Rarefaction curves showed that bacterial groups are more diverse than archaeal groups while the overall prokaryotic biodiversity is lower in high metal impacted environments than in less polluted habitats. Knowledge of this microbial community structure will help in understanding prokaryotic diversity, biogeography, and the role of microorganisms in zinc smelting AMD generation and perhaps it may be exploited for environmental remediation procedures in this area.     

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.     

Measurement of bacterial sulfate reduction in sediments: Evaluation of a single-step chromium reduction method

A procedure which includes the Total Reduced Inorganic Sulfur (TRIS) in a single distillation step is described for the radiotracer measurement of sulfate reduction in sediments. The TRIS includes both Acid Volatile Sulfide (AVS: H₂S + FeS) and the remaining Chromium Reducible Sulfur (CRS: S⁰, FeS₂). The single-step distillation was simpler and faster than the consecutive distillations of AVS and CRS. It also resulted in higher (4–50%) sulfate reduction rates than those obtained from the sum of³⁵S in AVS and CRS. The difference was largest when the sediment had been dried after AVS but before CRS distillation. Relative to the³⁵S-AVS distillation alone, the³⁵S-TRIS single-step distallation yielded 8–87% higher reduction rates. The separation and recovery of FeS, S⁰ and FeS₂ was studied under three distillation conditions: 1) cold acid, 2) cold acid with Cr²⁺, and 3) hot acid with Cr²⁺. The FeS was recovered by cold acid alone while pyrite was recovered by cold acid with Cr²⁺. A smaller S⁰ fraction, presumably of the finer crystal sizes, was recovered also in the cold acid with Cr²⁺ while most of the S⁰ required hot acid with Cr²⁺ for reduction to H₂S.     

In situ stimulation of bacterial sulfate reduction in sulfate-limited freshwater lake sediments

Abstract By adding sulfate in the form of solid gypsum, it was possible to transform in situ a predominantly methanogenic sediment ecosystem into a sulfate-reducing one. The concentrations of sulfate, sulfide, methane, acetate, propionate, soluble iron, and manganese were determined in the porewater before and after the transition. Although sulfate was no longer limiting, acetate and propionate continued to accumulate and reached much higher concentrations than under sulfate-limited conditions. Metabolic activities of fermenting bacteria and of sulfate reducers, which belong to the group that incompletely oxidizes organic material, might be responsible for the increased production of volatile fatty acids. The elevated concentrations of soluble Fe(II)²⁺ and Mn(II)²⁺ observed in the porewater stem from iron and manganese compounds which may be reduced chemically by hydrogen sulfide and other microbially produced reducing agents or directly through increased activities of the iron and manganese reducing bacteria. In the horizon with high sulfate-reducing activities the methane concentrations in the porewater were lower than in non-stimulated sediment regions. The shape of the concentration depth profile indicates methane consumption through sulfate reducing processes. The in situ experiment demonstrates the response of a natural microbial ecosystem to fluctuations in the environmental conditions.     

Adsorptive removal of copper and nickel ions from water using chitosan coated PVC beads

A new biosorbent was developed by coating chitosan, a naturally and abundantly available biopolymer, on to polyvinyl chloride (PVC) beads. The biosorbent was characterized by FTIR spectra, porosity and surface area analyses. Equilibrium and column flow adsorption characteristics of copper(II) and nickel(II) ions on the biosorbent were studied. The effect of pH, agitation time, concentration of adsorbate and amount of adsorbent on the extent of adsorption was investigated. The experimental data were fitted to Langmuir and Freundlich adsorption isotherms. The data were analyzed on the basis of Lagergren pseudo first order, pseudo-second order and Weber-Morris intraparticle diffusion models. The maximum monolayer adsorption capacity of chitosan coated PVC sorbent as obtained from Langmuir adsorption isotherm was found to be 87.9 mg g(-1) for Cu(II) and 120.5 mg g(-1) for Ni(II) ions, respectively. In addition, breakthrough curves were obtained from column flow experiments. The experimental results demonstrated that chitosan coated PVC beads could be used for the removal of Cu(II) and Ni(II) ions from aqueous medium through adsorption.     

Antiquity and evolutionary status of bacterial sulfate reduction: sulfur isotope evidence

The presently available sedimentary sulfur isotope record for the Precambrian seems to allow the following conclusions: (1) In the Early Archaean, sedimentary delta 34S patterns attributable to bacteriogenic sulfate reduction are generally absent. In particular, the delta 34S spread observed in the Isua banded iron formation (3.7 x 10(9) yr) is extremely narrow and coincides completely with the respective spreads yielded by contemporaneous rocks of assumed mantle derivation. Incipient minor differentiation of the isotope pattersn notably of Archaean sulfates may be accounted for by photosynthetic sulfur bacteria rather than by sulfate reducers. (2) Isotopic evidence of dissimilatory sulfate reduction is first observed in the upper Archaean of the Aldan Shield, Siberia (approximately 3.0 x 10(9) yr) and in the Michipicoten and Woman River banded iron formations of Canada (2.75 x 10(9) yr). This narrows down the possible time of appearance of sulfate respirers to the interval 2.8--3.1 x 10(9) yr. (3) Various lines of evidence indicate that photosynthesis is older than sulfate respiration, the SO4(2-) Utilized by the first sulfate reducers deriving most probably from oxidation of reduced sulfur compounds by photosynthetic sulfur bacteria. Sulfate respiration must, in turn, have antedated oxygen respiration as O2-respiring multicellular eucaryotes appear late in the Precambrian. (4) With the bulk of sulfate in the Archaean oceans probably produced by photosynthetic sulfur bacteria, the accumulation of SO4(2-) in the ancient seas must have preceded the buildup of appreciable steady state levels of free oxygen. Hence, the occurrence of sulfate evaporites in Archaean sediments does not necessarily provide testimony of oxidation weathering on the ancient continents and, consequently, of the existence of an atmospheric oxygen reservoir.     

Antiquity and evolutionary status of bacterial sulfate reduction: Sulfur isotope evidence

The presently available sedimentary sulfur isotope record for the Precambrian seems to allow the following conclusions: (1) In the Early Archaean, sedimentary ^{3 4} patterns attributable to bacteriogenic sulfate reduction are generally absent. In particular, the ^{3 4} spread observed in the Isua banded iron formation (3.7×10⁹ yr) is extremely narrow and coincides completely with the respective spreads yielded by contemporaneous rocks of assumed mantle derivation. Incipient minor differentiation of the isotope patterns notably of Archaean sulfates may be accounted for by photosynthetic sulfur bacteria rather than by sulfate reducers. (2) Isotopic evidence of dissimilatory sulfate reduction is first observed in the upper Archaean of the Aldan Shield, Siberia (3.0×10⁹ yr) and in the Michipicoten and Woman River banded iron formations of Canada (2.75×10⁹ yr). This narrows down the possible time of appearance of sulfate respirers to the interval 2.8–3.1×10⁹ yr. (3) Various lines of evidence indicate that photosynthesis is older than sulfate respiration, the SO ₄ ^2– utilized by the first sulfate reducers deriving most probably from oxidation of reduced sulfur compounds by photosynthetic sulfur bacteria. Sulfate respiration must, in turn, have antedated oxygen respiration as O₂-respiring multicellular eucaryotes appear late in the Precambrian. (4) With the bulk of sulfate in the Archaean oceans probably produced by photosynthetic sulfur bacteria, the accumulation of SO ₄ ^2– in the ancient seas must have preceded the buildup of appreciable steady state levels of free oxygen. Hence, the occurrence of sulfate evaporites in Archaean sediments does not necessarily provide testimony of oxidation weathering on the ancient continents and, consequently, of the existence of an atmospheric oxygen reservoir.Paper presented at the Fourth College Park Colloquium on Chemical Evolution, Limits of Life, October 18–20, 1978.     

水热法改性粉煤灰去除矿山酸性废水中金属离子

矿山酸性废水污染是由开采矿山和废弃矿山的所产生的重要环境问题。本文利用廉价的粉煤灰通过水热法改性制备了沸石吸附剂,研究了不同pH值、接触时间和吸附剂加入量条件下,矿山废水中的重金属离子去除效果。结果表明：pH值是影响重金属离子去除的重要因素,pH值在为7时,其对Pb²⁺、Zn²⁺、Cu²⁺去除率分别为100%、95.2%和95.8%;吸附剂对Cu²⁺的去除主要是由吸附和沉淀共同作用完成的,而Zn²⁺和Pb²⁺的去除主要依靠吸附作用。随着吸附剂加入量的增加,Pb²⁺、Zn²⁺、Cu²⁺的吸附率也随之增加,其最优加入量分别为20、25和30 g·L^-1。当Pb²⁺、Zn²⁺、Cu²⁺的吸附时间分别为20、40和50 min时,其去除率可分别可达94.2%、95.2%和98.2%。此外,还讨论了复合离子相互间强化和抑制作用。当3种离子同时存在时,其相互间的强化作用并不明显,但会呈现较显著的抑制作用。     

Lead and nickel removal using Microspora and Lemna minor

Aquatic plants can remove heavy metal contamination from the surrounding water. This study examined the ability of Microspora (a macro-alga) and Lemna minor (an aquatic plant) to remove soluble lead and nickel under various laboratory conditions. Microspora was tested in a batch and semi-batch process for lead removal. L. minor was tested in a batch process with lead and nickel to examine the potential competition between metals for adsorption. The Microspora was exposed to 39.4 mg/l of lead over 10 days. Results show up to 97% of the lead was removed in the batch process and 95% in the semi-batch process. Initial concentrations below 50 mg/l (a dose that kills the algae) had no effect on the final concentration. The L. minor was exposed to lead and nickel using a full 3(2) factorial experimental design (nine experiments, plus replications). Initial lead concentrations were 0.0, 5.0, and 10.0 mg/l, and nickel concentrations were 0.0, 2.5, and 5.0 mg/l in the experiment. Overall, L. minor removed 76% of the lead, and 82% of the nickel. No synergistic/antagonistic effect was noted for the multiple metal experiments, in terms of metal removal.     

Nickel removal from nickel plating waste water using a biologically active moving-bed sand filter

Efficient removal of dissolved nickel was observed in a biologically active moving-bed `MERESAFIN' sand filter treating rinsing water from an electroless nickel plating plant. Although nickel is fully soluble in this waste water, its passage through the sand filter promoted rapid removal of approximately 1 mg Ni/l. The speciation of Ni in the waste water was modelled; the most probable precipitates forming under the conditions in the filter were predicted using PHREEQC. Analyses of the Ni-containing biosludge using chemical, electron microscopical and X-ray spectroscopic techniques confirmed crystallisation of nickel phosphate as arupite (Ni₃(PO₄)₂.8H₂O), together with hydroxyapatite within the bacterial biofilm on the filter sand grains. Biosorption contributed less than 1% of the overall sequestered nickel. Metabolising bacteria are essential for the process; the definitive role of specific components of the mixed population is undefined but the increase in pH promoted by metabolic activity of some microbial components is likely to promote nickel desolubilisation by others.