A comparative study on metal sorption by brown seaweed

This study compared the sorption of Ag, Cd, Co, Cd, Mn, Ni, Pb and Zn by a Ca-treated Sargassum biomass at pH 5.0, under low and high ionic strength (IS) conditions. The sorption isotherms of As [As(V)] and Cr [Cr(III) and Cr(VI)] were also determined at low IS. The isotherm data for the eight cationic metals and Cr(III) were well fitted by Langmuir equations. Generally, the maximum metal uptake (Umax) followed: Cr(III) > Pb approximately Cu > Ag approximately Zn approximately Cd > Ni approximately Mn approximately Co > Cr(VI) > As(V) at low IS and Pb > Cu > Co > Mn approximately Cd > Zn approximately Ag > Ni at high IS. As(V) did not bind to the seaweed at pH 5.0. The results indicated that sorption of Pb was not affected by the increasing IS, though the percentage of free Pb ions in the water was greatly reduced as predicted by the speciation model. High IS lowered Umax by 10-36% (except Co and Pb), and lowered the affinity constant of the metal by 33-91% for all cationic metals, as compared to low IS. Moreover, the removal efficiency of the cationic metals and Cr decreased exponentially with initial metal concentrations and was lower at high IS. Ion-exchange was the mechanism responsible for the cationic metal sorption onto the seaweed, and Na ion interfered with the cationic metal binding through electrostatic interaction. In conclusion, this study showed the differential binding capacity of the Sargassm biomass for different metals and oxidation states and the differential effects of IS. According to the present results, Sargassum may be considered a good biosorbent for cationic metals (especially Pb) in both low and high-salt containing wastewater.     

Aseptic hydroponics to assess rhamnolipid-Cd and rhamnolipid-Zn bioavailability for sunflower (<Emphasis Type="Italic">Helianthus annuus</Emphasis>): a phytoextraction mechanism study

The availability of cadmium (Cd) and zinc (Zn) to sunflower (Helianthus annuus) was investigated in rhamnolipid- and ethylenediaminetetraacetic acid (EDTA)-buffered solutions in order to evaluate the influence of aqueous speciation of the metals on their uptake by the plant, in relation to predictions of uptake by the free ion activity model (FIAM). Free metal ion activity was estimated using the chemical equilibrium program MINTEQ or measured by Donnan dialysis. The uptake of Cd followed the FIAM for the EDTA-buffered solution at EDTA concentrations below 0.4 μM; for the rhamnolipid-buffered solution, the uptake of both metals in roots was not markedly affected by increasing rhamnolipid concentrations in solution. This suggests rhamnolipid enhanced metal accumulation in plant roots (per unit free metal in solution) possibly through formation and uptake of lipophilic complexes. The addition of normal Ca concentrations (low millimetre range) to the rhamnolipid uptake solutions reduced Cd accumulation in shoots by inhibiting Cd translocation, whereas it significantly increased Zn accumulation in shoots. This study confirms that although rhamnolipid could enhance accumulation of Cd in plants roots at low Ca supply, it is not suitable for Cd phytoextraction in contaminated soil environments where Ca concentrations in soil solution are orders of magnitude greater than those of Cd.     

Influences of salinity on the biokinetics of Cd, Se, and Zn in the intertidal mudskipper Periophthalmus cantonensis

The biokinetics (aqueous uptake, dietary assimilation, and elimination) of Cd, Se, and Zn in the intertidal mudskipper, Periophthalmus cantonensis, were examined at different acclimated salinities using the radiotracer technique. The dietary assimilation efficiency from ingested radiolabeled polychaetes was the highest for Se (32–40%), followed by Zn (5–7%) and Cd (2–3%), and was not influenced by salinity within a range of 10–30 psu. Uptake from the dissolved phase typically exhibited a linear pattern within the first 12 h of exposure, followed by a second slower uptake. The highest concentration factor (CF) was found for Zn, followed by Cd and Se. Differences in salinity did not significantly affect the CF of the three metals within the first 12 h, but the CFs were significantly higher at lower salinities (10–20 psu) than at the highest salinity (30 psu) by the end of 48 h exposure. Because the degrees to which the uptake of Se (a metalloid) and Cd/Zn were affected by salinity were comparable, we concluded that metal speciation as a result of salinity change was not important in leading to a change in metal CF. Physiological changes may be responsible for the increasing uptake at lowered salinity. The elimination rates of the three metals (0.01–0.06 d⁻¹) were not significantly affected by salinity, but Se was eliminated at a faster rate following aqueous uptake than following dietary ingestion. There was no consistent influence of exposure routes on Cd and Zn elimination. The accumulated Cd was mainly associated with the gut, whereas the muscle was the dominant target site for Se and Zn accumulation.     

Effects of major nutrient additions on metal uptake in phytoplankton

We examined the influences of major nutrients (N, P, Si) on the accumulation of three trace metals [Cd, Se(IV), and Zn] in four species of marine phytoplankton (diatom, green alga, dinoflagellate, prasinophyte). Relative metal uptake was quantified by the kinetic measurements of metal concentration factor over a short exposure period. Our study demonstrated that nutrient addition significantly influenced the metal uptake rate and the cell growth rate in all four phytoplankton species. An increase in ambient N concentration considerably enhanced metal uptake by the cells. The dry weight concentration factor increased by 2.4-14.9 times for Cd, 1.1-4.0 times for Se, and 1.1-5.4 times for Zn in all four phytoplankton species with an addition of 176.4 microM N. The effects of P or Si addition on metal uptake and cell growth were less pronounced than the effects of N addition. Under most circumstances the rate of metal uptake increased exponentially with increasing cell growth rate constant. Only Se(IV) uptake in the diatom Phaeodactylum tricornutum was not correlated with cell growth rate. Se(IV) was not accumulated by the green algae Chlorella autotrophica at a high P concentration (7.2 microM), but appreciable accumulation was documented in cells inoculated without P addition. Our study therefore demonstrated that nutrient enrichments in many coastal waters can considerably affect trace metal uptake in phytoplankton and presumably metal trophic transfer in marine food chains.     

Metal uptake capacity of modified Saccharomyces pastorianus biomass from different types of solution

In this paper, we investigate the effect of different biomass pretreatments on metal ion uptake by various biosorbents. Heat-treated as well as caustic-treated and ground biomass of Saccharomyces pastorianus was used to remove copper, lead and cadmium from various solutions. Untreated yeast was used as the control sample. The effect of yeast modification on sorption capacity depended on the different types of heavy metal ions and whether they were in single- or multi-component solutions. The highest uptake of copper and lead from a single-metal solution was obtained from heat-treated cells. Ground biomass was the most efficient at cadmium removal. However, the sorption capacity of the modified biomass did not improve when metal ions were removed from multi-component solutions. Indeed, the results in this paper show that optimizing metal removal from single-cation solutions can lead to decreased sorption capacity in multi-component solutions. Therefore, while adjusting the procedure of biomass modification, not only the nature of the metal ion being sorbed but also the chemical composition of the metal ion solution should be taken into account.     

Sorption kinetics and leachability of heavy metal from the contaminated soil amended with immobilizing agent (humus soil and hydroxyapatite)

Release of heavy metals onto the soil as a result of agricultural and industrial activities may pose a serious threat to the environment. This study investigated the kinetics of sorption of heavy metals on the non-humus soil amended with (1:3) humus soil and 1% hydroxyapatite used for in situ immobilization and leachability of heavy metals from these soils. For this, a batch equilibrium experiment was performed to evaluate metal sorption in the presence of 0.05 M KNO(3) background electrolyte solutions. The Langmuir isotherms applied for sorption studies showed that the amount of metal sorbed on the amended soil decreased in the order of Pb(2+)>Zn(2+)>Cd(2+). The data suggested the possibility of immobilization of Pb due to sorption process and immobilization of Zn and Cd by other processes like co-precipitation and ion exchange. The sorption kinetics data showed the pseudo-second-order reaction kinetics rather than pseudo-first-order kinetics. Leachability study was performed at various pHs (ranging from 3 to 10). Leachability rate was slowest for the Pb(2+) followed by Zn(2+) and Cd(2+). Out of the metal adsorbed on the soil only 6.1-21.6% of Pb, 7.3-39% of Zn and 9.3-44.3% of Cd leached out from the amended soil.     

Removal of Cu and Pb by tartaric acid modified rice husk from aqueous solutions

A study on the modification of rice husk by various carboxylic acids showed that tartaric acid modified rice husk (TARH) had the highest binding capacities for Cu and Pb. The carboxyl groups on the surface of the modified rice husk were primarily responsible for the sorption of metal ions. A series of batch experiments using TARH as the sorbent for the removal of Cu and Pb showed that the sorption process was pH dependent, rapid and exothermic. The sorption process conformed to the Langmuir isotherm with maximum sorption capacities of 29 and 108 mg/g at 27 +/- 2 degrees C for Cu and Pb, respectively. The uptake increased with agitation rate. Decrease in sorbent particle size led to an increase in the sorption of metal ions and this could be explained by an increase in surface area and hence binding sites. Metal uptake was reduced in the presence of competitive cations and chelators. The affinity of TARH for Pb is greater than Cu.     

Effect of clays, metal oxides, and organic matter on rhamnolipid biosurfactant sorption by soil

Rhamnolipids produced by Pseudomonas aeruginosa have been proposed as soil washing agents for enhanced removal of metal and organic contaminants from soil. A potential limitation for the application of rhamnolipids is sorption by soil matrix components. The objective of this study is to empirically determine the contribution of representative soil constituents (clays, metal oxides, and organic matter) to sorption of the rhamnolipid form most efficient at metal complexation (monorhamnolipid). Sorption studies show that monorhamnolipid (R1) sorption is concentration dependent. At low R1 concentrations that are relevant for enhancing organic contaminant biodegradation, R1 sorption followed the order: hematite (Fe(2)O(3))>kaolinite>MnO(2) approximately illite approximately Ca-montmorillonite>gibbsite (Al(OH)(3))>humic acid-coated silica. At high R1 concentrations, relevant for use in complexation/removal of metals or organics, R1 sorption followed the order: illite>humic acid-coated silica>Ca-montmorillonite>hematite>MnO(2)>gibbsite approximately kaolinite. These results allowed prediction of R1 sorption by a series of six soils. Finally, a comparison of R1 and R2 (dirhamnolipid) shows that the R1 form sorbs more strongly alone than when in a mixture of both the R1 and R2 forms. The information presented can be used to estimate, on an individual soil basis, the extent of rhamnolipid sorption. This is important for determining: (1) whether rhamnolipid addition is a feasible remediation option and (2) the amount of rhamnolipid required to efficiently remove the contaminant.     

Solanum nigrum grown in contaminated soil: effect of arbuscular mycorrhizal fungi on zinc accumulation and histolocalisation

Zn tissue accumulation in Solanum nigrum grown in a non-contaminated and a naturally contaminated Zn matrix and the effect of inoculation with different arbuscular mycorrhizal fungi (AMF) on metal uptake were assessed. S. nigrum grown in the contaminated soil always presented higher Zn accumulation in the tissues, accumulating up to 1622 mg Zn kg(-1). The presence of both Glomus claroideum and Glomus intraradices enhanced the uptake and accumulation of Zn by S. nigrum (up to 83 and 49% higher Zn accumulation, respectively). The main deposits of the metal were found in the intercellular spaces and in the cell walls of the root tissues, as revealed by autometallography, with the inoculation with different AMF species causing no differences in the location of Zn accumulation. These findings indicate that S. nigrum inoculated with selected heavy metal tolerant AMF presents extracting and accumulating capacities, constituting a potentially suitable remediation method for Zn polluted soils.     

Metal accumulation in the smooth toadfish, Tetractenos glaber, in estuaries around Sydney, Australia

This study determined the metal levels in sediments and tissues of a common estuarine fish, Tetractenos glaber (smooth toadfish), from two metal contaminated and two reference estuaries near Sydney, Australia. Metal levels were highest in sediments and fish from contaminated estuaries. Gonads contained the highest metal levels followed by muscle, gill and liver. Metal accumulation was gender-dependant (e.g. male gonads were >20 times higher in As than females; female gills were >30 times higher than males for Pb). Cadmium, Pb and Ni levels in fish tissues reflected sediment levels, indicating sediment and/or dietary metal uptake. Levels of As, Co, Cd & Pb in gills showed similar patterns to other tissues, suggesting that metals may have been taken up by gills through contaminated water. Similar metal patterns in tissues and sediments suggest more than one uptake pathway. This study indicates that multiple factors influence metal accumulation in fish.     

Mechanisms of lead, copper, and zinc retention by phosphate rock

The solid-liquid interface reaction between phosphate rock (PR) and metals (Pb, Cu, and Zn) was studied. Phosphate rock has the highest affinity for Pb, followed by Cu and Zn, with sorption capacities of 138, 114, and 83.2 mmol/kg PR, respectively. In the Pb-Cu-Zn ternary system, competitive metal sorption occurred with sorption capacity reduction of 15.2%, 48.3%, and 75.6% for Pb, Cu, and Zn, respectively compared to the mono-metal systems. A fractional factorial design showed the interfering effect in the order of Pb>Cu>Zn. Desorption of Cu and Zn was sensitive to pH change, increasing with pH decline, whereas Pb desorption was decreased with a strongly acidic TCLP extracting solution (pH = 2.93). The greatest stability of Pb retention by PR can be attributed to the formation of insoluble fluoropyromorphite [Pb(10)(PO(4))(6)F(2)], which was primarily responsible for Pb immobilization (up to 78.3%), with less contribution from the surface adsorption or complexation (21.7%), compared to 74.5% for Cu and 95.7% for Zn. Solution pH reduction during metal retention and flow calorimetry analysis both supported the hypothesis of retention of Pb, Cu, and Zn by surface adsorption or complexation. Flow calorimetry indicated that Pb and Cu adsorption onto PR was exothermic, while Zn sorption was endothermic. Our research demonstrated that PR can effectively remove Pb from solutions, even in the presence of other heavy metals (e.g. Cu, Zn).     

Sorption of Cu, Pb and Cr on Na-montmorillonite: competition and effect of major elements

The competitive sorption among Cu, Pb and Cr in ternary system on Na-montmorillonite at pH 3.5, 4.5 and 5.5 and at different heavy metal concentrations, and the effect of varying concentrations of Al, Fe, Ca and Mg on the sorption of heavy metals were studied. Competitive sorption of Cu, Pb and Cr in ternary system on montmorillonite followed the sequence of Cr ? Cu > Pb. Moreover, the competition was weakened by the increase of pH while was intensified by the increase of heavy metal concentration. The sorption of heavy metal on montmorillonite was inhibited by the presence of Ca and Mg, while Al and Fe showed different patterns in affecting heavy metal sorption. Aluminum and Fe generally inhibited the sorption of heavy metal when the pH and/or concentration of major elements were relatively low. However, promoting effects on heavy metal sorption by Al and Fe were found at relatively high pH and/or great concentration of major elements. The inhibition of major elements on heavy metal sorption generally followed the order of Al > Fe > Ca ? Mg, while Fe was more efficient than Al in promoting the sorption of heavy metals. These findings are of fundamental significance for evaluating the mobility of heavy metals in polluted environments.     

Seizure modeling of Pb(II) and Cd(II) from aqueous solution by chemically modified sugarcane bagasse fly ash: isotherms, kinetics, and column study

Heavy metal pollution is a common environmental problem all over the world. The purpose of the research is to examine the applicability of bagasse fly ash (BFA)—an agricultural waste of sugar industry used for the synthesis of zeolitic material. The zeolitic material are used for the uptake of Pb(II) and Cd(II) heavy metal. Bagasse fly ash is used as a native material for the synthesis of zeolitic materials by conventional hydrothermal treatment without (conventional zeolitic bagasse fly ash (CZBFA)) and with electrolyte (conventional zeolitic bagasse fly ash in electrolyte media (ECZBFA)) media. Heavy metal ions Pb(II) and Cd(II) were successfully seized from aqueous media using these synthesized zeolitic materials. In this study, the zeolitic materials were well characterized by different instrumental methods such as Brunauer–Emmett–Teller, XRF, Fourier transform infrared spectroscopy, powder X-ray diffraction, and scanning electron microscopic microphotographs. The presence of analcime, phillipsite, and zeolite P in adsorbents confirms successful conversion of native BFA into zeolitic materials. Seizure modeling of Pb(II) and Cd(II) was achieved by batch sorption experiments, isotherms, and kinetic studies. These data were used to compare and evaluate the zeolitic materials as potential sorbents for the uptake of heavy metal ions from an aqueous media. The Langmuir isotherm correlation coefficient parameters best fit the equilibrium data which indicate the physical sorption. Pseudo-second-order and intra-particle diffusion model matches best which indicates that the rate of sorption was controlled by film diffusion. The column studies were performed for the practical function of sorbents, and breakthrough curves were obtained, which revealed higher sorption capacity as compared to batch method. Synthesized zeolitic material (CZBFA and ECZBFA), a low-cost sorbent, was proven as potential sorbent for the uptake of Pb(II) and Cd(II) heavy metal ions.     

Stabilization of the initial electrochemical potential for a metal-based potentiometric titration study of a biosorption process

An interactive metal-based potentiometric titration method has been developed using an ion selective electrode for studying the sorption of metal cations. The accuracy of this technique was verified by analyzing the metal sorption mechanism for the biomass of Rhizopus arrhizus fungus and diatomite, two dissimilar materials (organic and mineral, strong sorbent and weak sorbent) of a different order of cation exchange capacity. The problem of the initial electrochemical potential was addressed identifying the usefulness of a Na-sulfonic resin as a strong chelating agent applied before the beginning of sorption titration experiments so that the titration curves and the sorption uptake could be quantitatively compared. The resin stabilized the initial electrochemical potential to -405+/-5 mV corresponding to 2 micro gl(-1) of lead concentration in solution. The amounts of lead sorbed by R. arrhizus biomass and diatomite were 0.9 mmol g(-1) (C(e)=5.16 x 10(-2)mM) and 0.052 mmol g(-1) (C(e)=5.97 x 10(-2) mM), respectively. Lead sorption by the fungal biomass was pinpointed to at least two types of chemical active sites. The first type was distinguished by high reactivity and a low number of sites whereas the other was characterized by their higher number and lower reactivity.     

The marine macroalga Cystoseira baccata as biosorbent for cadmium(II) and lead(II) removal: kinetic and equilibrium studies

This work reports kinetic and equilibrium studies of cadmium(II) and lead(II) adsorption by the brown seaweed Cystoseira baccata. Kinetic experiments demonstrated rapid metal uptake. Kinetic data were satisfactorily described by a pseudo-second order chemical sorption process. Temperature change from 15 to 45 degrees C showed small variation on kinetic parameters. Langmuir-Freundlich equation was selected to describe the metal isotherms and the proton binding in acid-base titrations. The maximum metal uptake values were around 0.9 mmol g(-1) (101 and 186 mg g(-1) for cadmium(II) and lead(II), respectively) at pH 4.5 (raw biomass), while the number of weak acid groups were 2.2 mmol g(-1) and their proton binding constant, K(H), 10(3.67) (protonated biomass). FTIR analysis confirmed the participation of carboxyl groups in metal uptake. The metal sorption was found to increase with the solution pH reaching a plateau above pH 4. Calcium and sodium nitrate salts in solution were found to affect considerably the metal biosorption.     

Uptake and distribution of Zn, Cu, Cd, and Pb in an aquatic plant Potamogeton natans

A better understanding of metal uptake and translocation by aquatic plants can be used to enhance the performance of constructed wetland systems for stormwater treatment. Specifically, this study examines whether the uptake of Zn, Cu, Cd, and Pb by Potamogeton natans is via the leaves, stems, or roots, and whether there is translocation from organs of uptake to other plant parts. Competition between the metals at uptake and at the level of the cell wall-bound part of the metals accumulated in stem and leaf tissue was also examined. The results show that Zn, Cu, Cd, and Pb were taken up by the leaves, stems, and roots, with the highest accumulation found in the roots. At the elevated metal concentrations common in stormwater the uptake of Cu, but not of Zn, Cd, or Pb, by the roots was somewhat limited at uptake due to competition with other metals. Between 24% and 59% of the metal content was bound to the cell walls of the plant. Except in the case of Pb, the cell wall-bound fraction was generally smaller in stems than in leaves. No translocation of the metals to other parts of the plant was found, except for Cd which was translocated from leaf to stem and vice versa. Dispersion of metals from sediment to water through P. natans is therefore unlikely.     

Phytoremediation potential of Spirulina (Arthrospira) platensis: biosorption and toxicity studies of cadmium

This study examines the possibility of using Spirulina (Arthrospira) platensis TISTR 8217 to remove low concentrations of cadmium (less than 100 mg/l) from wastewater. The cyanobacteria were exposed to six different cadmium concentrations for 96 h, and the growth rate was determined using an optical density at 560 nm. The inhibiting concentration (IC50) was estimated using probit analysis. The IC50 at 24, 48, 72, and 96 h were 13.15, 16.68, 17.28, and 18.35 mg/l Cd, respectively. Cellular damage was studied under a light microscope and a transmission electron microscope. Swollen cells and fragmented filaments were observed. Cell injury increased with increasing concentrations of cadmium. Ultrastructural changes were observed in the algae exposed to cadmium concentrations both close to IC50 (14.68 mg/l) and at IC50 (18.35 mg/l). The alterations induced by cadmium were disintegration and disorganization of thylakoid membranes, presence of large intrathylakoidal space, increase of polyphosphate bodies, and cell lysis. In addition, the cadmium adsorption by algal cells was studied. Environmental factors were found to have an effect on biosorption. The uptake of cadmium was not affected by the temperature of the solution, but the sorption was pH dependent. The optimum pH for biosorption of algal cells was 7. The cadmium uptake process was rapid, with 78% of metal sorption completed within 5 min. The sorption data fit well to the Langmuir isotherm. The maximum adsorption capacity for S. platensis was 98.04 mg Cd per g biomass.     

Modelling the extra and intracellular uptake and discharge of heavy metals in Fontinalis antipyretica transplanted along a heavy metal and pH contamination gradient

Samples of the aquatic bryophyte Fontinalis antipyretica Hedw. were transplanted to different sites with the aim of characterizing the kinetics of the uptake and discharge of heavy metals in the extra and intracellular compartments. The accumulation of metals in extracellular compartments, characterized by an initial rapid accumulation, then a gradual slowing down over time, fitted perfectly to a Michaelis-Menten model. The discharge of metals from the same compartment followed an inverse linear model or an inverse Michaelis-Menten model, depending on the metal. In intracellular sites both uptake and discharge occurred more slowly and progressively, following a linear model. We also observed that the acidity of the environment greatly affected metal accumulation in extracellular sites, even when the metals were present at relatively high concentrations, whereas the uptake of metals within cells was much less affected by pH.     

Biosorption of arsenic in drinking water by submerged plant: Hydrilla verticilata

To evaluate the biosorption efficacy of submerged aquatic plant Hydrilla verticilata for arsenic uptake from drinking water. H. verticillata, a submerged aquatic plant was utilized successfully for arsenic uptake from aqueous solution. Batch studies with various parameters viz. pH, sorbent dose, contact time, initial metal ion concentration, and temperature were carried out. Data were utilized to plot Lagergren graph along with pseudo-second-order graphs for kinetic studies to estimate the removal efficacy and to determine the nature of reaction. Scanning electron microscopy (SEM) and Fourier transform infrared spectroscopy (FTIR) have been performed for characterization of metals on biomass. The study showed 96.35 % maximum absorption of arsenic by H. verticilata at initial concentration of 100 ppb with 0.5 g of biomass/100 ml for 5 h contact time at pH?6.0 with 150 rpm agitation rate. Data followed Langmuir isotherm showing sorption to be monolayer on homogeneous surface of biosorbent. The negative values of ΔG° indicated spontaneous nature; whereas ΔH° indicates exothermic nature of system and negative value of ?S° entropy change correspond to a decrease in the degree of freedom to the adsorbed species followed by pseudo-second-order adsorption kinetics. FTIR and SEM results showed apparent changes in functional group regions after metal chelation and the changes in surface morphology of biosorbent. This is a comparatively more effective, economic, easily available, and environmentally safe source for arsenic uptake from solution due to its high biosorption efficacy than other biosorbents already used.     

An insight into the mechanism of biosorption of copper by Bacillus polymyxa

Our main objective was to isolate bacteria from soil contaminated with effluents from electroplating wastewater and assess their potential to remove copper from the aqueous phase. The strain identified as Bacillus polymyxa accumulated copper inside the cell, where it bound to the cell wall. The intracellular metal accumulation led to the inhibition of dehydrogenases, which is essential for the energy deriving reactions. Addition of thiol group containing compounds, like reduced glutathione, dithiothreitol and mercapto-ethanol, revived the enzyme activity, implicating the sulfhydryl groups as the target of metal action. However, when the cells were exposed to higher concentration of Cu(II), irreversible enzyme denaturation occurred. Desorption with DTT and citrate showed that 82% of the metal was accumulated on cell surface and the contribution from metabolism-dependent intracellular accumulation was only 10-12%. The surface characterization of B. polymyxa indicated the presence of carboxyl, imidazolium, amino and phenolic groups, which might be responsible for metal uptake. Electron paramagnetic resonance (EPR) spectroscopy studies indicated that the metal coordinating environment could be either CuN₂O₂ or CuNO₃ chromophores, which may be the result of binding of Cu(II) to oxygen atoms of carboxylic groups of cell wall peptidoglycan and nitrogen atoms of amino-sugars or structural proteins. This environment got slightly altered after several treatments of B. polymyxa, leading to the formation of CuO₄, suggesting the coordination of copper to four oxygen atoms derived from carboxyl groups of peptidoglycan.