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Environ. Sci. Technol.

2000, 34, 1081-1087

Environment Act, increases in landfill tax. and a commitment


An Integrated Method Incorporating by the U.K. government to build 60% of new houses on
Sulfur-Oxidizing Bacteria and previously developed sites (2) have increased the pressure
to develop effective remediation technologies. Over 60% of
Electrokinetics To Enhance Removal sites are co-contaminated with metals and organics. The
metals are frequently found as complex mixtures. and many
of Copper from Contaminated Soil former industrial sites contain highly toxic metals such as
cadmium, nickel. and arsenic. Therefore, in the U.K., as in
GIACOMO MAINI,§ AJAY K. SHARMAN,§ other countries, the potential for developing these sites
GARRY SUNDERLAND,‡ together with the possibility that metals may leach into
CHRISTOPHER. J. KNOWLES,† AND groundwater or enter the food chain through plant material
S I M O N A . J A C K M A N * ,† means that remediation of metal-contaminated land is a
IBS-Viridian Ltd., 114-116 John Wilson Business Park, priority. There are currently few comprehensive remediation
Thanet Way, Whitstable, Kent CT5 3QT, U.K., EA Technology technologies available for such sites.
Ltd., Capenhurst, Chester CH1 6ES, U.K., Oxford Centre for Within the mining industries, technologies have been
Environmental Biotechnology, Department of Engineering developed for the recovery of valuable metals from ores and
Science, University of Oxford, Parks Road, Oxford OX1 3PJ, rock/soil materials. Indeed the use of bacteria in bioleaching
and NERC Institute of Virology & Environmental has become a prominent method of recovery (3). Naturally
Microbiology, Mansfield Road, Oxford OX1 3SR, U.K. occurring iron- and sulfur-oxidizing bacteria from mining
wastes have been isolated and strains selected for their ability
to solubilize metals from different substrates (4). In this
respect, the bacteria involved in bioleaching processes are
The combination of bioleaching and electrokinetics for able to convert metal sulfides to their respective sulfates,
the remediation of metal contaminated land has been thereby transforming them from insoluble to soluble salts.
investigated. In bioleaching, bacteria convert reduced sulfur The metals can be recovered by washing and further
compounds to sulfuric acid, acidifying soil and mobilizing processing. This established technology is now being ex-
metal ions. In electrokinetics, DC current acidifies soil, and tended to investigate the leaching of contaminating metals
mobilized metals are transported to the cathode by from soils as a bioremediation technique (5). The application
of mixed cultures of bacteria has proved successful in
electromigration. When bioleaching was applied to silt
mobilizing metals within different soils. Difficulties, however,
soil artificially contaminated with seven metals and amended may be presented by inhibition of microbial activity due to
with sulfur, bacterial activity was partially inhibited and the mobilized metals, with the potential for the entire
limited acidification occurred. Electrokinetic treatment of bioleaching process to come to a halt (6). The presence of
silt soil contaminated solely with 1000 mg/kg copper nitrate anionic species is also known to inhibit sulfur oxidation as
showed 89% removal of copper from the soil within 15 pH values are reduced (7). The potential for metals to leach
days. To combine bioleaching and electrokinetics sequentially, from soils means that to avoid groundwater contamination
preliminary partial acidification was performed by the process has either to be conducted ex situ, or, if in situ,
amending copper-contaminated soil with sulfur (to 5% with a metal removal system.
w/w) and incubating at constant moisture (30% w/w) and Electrokinetics is an emerging engineering technique for
temperature (20 °C) for 90 days. Indigenous sulfur- the remediation of contaminated land (8-10). The applica-
tion of a direct current to soil, by insertion of electrodes,
oxidizing bacteria partially acidified the soil from pH 8.1 to
leads to the generation of hydrogen ions at the anode and
5.4. This soil was then treated by electrokinetics yielding hydroxyl ions at the cathode. These migrate into the soil
86% copper removal in 16 days. In the combined process, and the hydrogen ions can displace adsorbed metal ions
electrokinetics stimulated sulfur oxidation, by removing into the pore fluid of the soil. By manipulation of the
inhibitory factors, yielding a 5.1-fold increase in soil sulfate conditions surrounding the cathode, an acidic pH can be
concentration. Preacidification by sulfur-oxidizing bacteria maintained throughout the soil. Metal ions, once solubilized,
increased the cost-effectiveness of the electrokinetic can be transported by electromigration through the soil and
treatment by reducing the power requirement by 66%. recovered at the cathode. The process has been effective in
both model and real systems and has been applied to sites
in the United States (11) and in Europe (12).
The combination of bioleaching and electrokinetics has
Introduction the potential to overcome several of the limitations of the
In the U.K. alone, 200 hundred years of industrialization has individual techniques with the possibility that some of the
resulted in an estimated 200 000 contaminated sites (1). A combined attributes may prove to be synergistic. In particular,
significant number of these contain cocktails of toxic electrokinetics mobilizes metal ions provided their speciation
chemicals that provide a hazard to both human health and is appropriate. Metals as hydroxides or oxides may be
the environment. The Confederation of British Industry has solubilized by the electrokinetic acidification, but those
recently estimated the cost of remediating this contaminated present as insoluble sulfides, a common speciation in former
land at £ 20bn (1). The implementation of the U.K. 1995 gasworks sites and mining wastes, will not be extracted by
this method. However, the bacteria involved in bioleaching
* Corresponding author phone: +44 1865 281630; fax: +44 1865 processes can convert metal sulfides to sulfates, thereby
281696; e-mail: saja@wpo.nerc.au.uk.
§ IBS-Viridian Ltd. enabling their solubilization and subsequent transport by
‡ EA Technology Ltd. electromigration. In addition, the directional transport of
† University of Oxford and NERC Institute of Virology & Envi- metal ions by electrokinetics is a useful complement to
ronmental Microbiology. bioleaching as solubilized metals can be removed at the
10.1021/es990551t CCC: $19.00  2000 American Chemical Society VOL. 34, NO. 6, 2000 / ENVIRONMENTAL SCIENCE & TECHNOLOGY 9 1081
Published on Web 02/17/2000
preliminary bioleaching experiments with static soil and soil
TABLE 1. Characteristics of Standardized Silt Soil slurries, it was decided to focus upon a single contaminating
parameter value metal for electrokinetic studies. This would enable a clearer
elucidation of the complementary effects of bioleaching and
pH 8.1 electrokinetic metal removal. Copper was chosen as the
soil density (kg/dm3) 1.350 contaminant and applied as 1000 mg/kg copper nitrate.
extractable phosphorus (mg/L) 24
particle size, diameter (mm) <2
Static Soil Experiment. Contaminated and uncontami-
cationic exchange capacity (me/100 g) 21.7 nated soils were amended with sulfur (mesh size 50) at two
soil buffering capacity (g/kg) (calcd as the 0.52 different concentrations (0.5 and 5% w/w) by slow addition
amount of [H+] from sulfuric acid of sulfur with constant mixing. Water was added to 30% (w/
required to acidify the soil to pH 2) w). Soils were set aside in trays with a soil depth of 10 cm
permeability, k (m/s) 1.81 × 10-11 at constant temperature (20 °C) and moisture (maintained
copper concn (mg/kg) 35.5 by regular dry weight measurement and addition of water)
calcium carbonate (%) 2 for up to 42 days. After 21 and 42 days, samples were taken
total sulfur (%) 0.08
and analyzed for pH (by suspending 1 g of soil in 10 mL of
total nitrogen (%) 0.25
organic matter (%) 3.40 water) and the concentration of sulfate. Sulfate analysis was
organic carbon (%) 1.97 by addition of 10 mL of 0.1 M HCl to 1 g of soil and mixing.
C:N ratio 7.9:1 Following centrifugation, barium chloride was added to
samples, precipitating barium sulfate, which was analyzed
by spectrophotometry.
cathode for straightforward downstream processing. A further Soil Slurry Experiment. Soil slurries were prepared from
advantage is the potential for preliminary bioleaching prior contaminated and uncontaminated silt soil amended with
to electrokinetics to reduce the overall time scale and cost sulfur and added to deionized water at 30% (w/v) to give a
of electrokinetic remediation. By amendment of the soil with total volume of 300 mL in 500 mL conical flasks. Slurries
inexpensive sulfur (below $ 250/tonne) and simply allowing were incubated with shaking (160 rpm) at 30 °C, and samples
the soil bacteria to metabolize, predictable sulfate generation were taken for analysis of pH and sulfate concentration.
and acidification can be achieved. Little intervention, in terms Electrokinetic Studies. The apparatus for performing
of man hours, is required in comparison to acid washing of electrokinetics is outlined in Figure 1. A soil cell system was
soil with relatively more expensive and hazardous chemicals constructed with six individual compartments each being
(e.g. sulfuric acid). This preliminary bioleaching stage forms 6.5 cm long between anode and cathode, 3 cm wide and 4
an important part of the current study. cm deep, and containing approximately 150 g of moist soil.
Electrokinetics together with bioremediation has been This arrangement allowed samples to be taken at a range of
used to effect the movement and degradation of phenol by time points. Within each compartment, soil could be sliced
an industrial/government consortium in the United States into five centimeter-wide sections to enable measurements
(13). The same consortium has developed this technology of pH, moisture content, and metal ions. pH was determined
up to a full-scale field test to move trichloroethylene (TCE) by suspending 1 g of soil in 10 mL of water and using a
into treatment zones in which it is degraded by reaction with standard pH meter. Moisture content was determined by
zerovalent iron (14). The introduction of nutrients into soil drying soil at 110 °C for 24 h and comparing wet and dry
by electrokinetics (15, 16) and movement of other organics weights. Metal ions in soil were analyzed by Electron
have also been investigated (17), but there has been little use Diffraction X-ray Fluorescence (ED-XRF; CPL Laboratories,
of combined techniques for remediation of metal contami- Derbyshire, U.K.). Anodes were of carbon felt material (8.5
nation. The behavior of sulfur-oxidizing bacteria at acidic cm wide, 7 cm deep, and 1.5 cm thick), and stainless steel
pH values has been well studied (18), and the acidophilic mesh (14 cm long and 5 cm wide folded in the center) was
bacterium Thiobacillus ferrooxidans has been used for used for cathodes. Each cathode was located in a compart-
combined electro/bioleaching processes for metal recovery ment (8.5 cm wide, 7 cm deep containing approximately 80
(19, 20). The paucity of data concerning microbial activities mL of liquid) bordered by a semipermeable membrane to
in soil in electric fields led us, therefore, to investigate the allow ions and water to pass from soil to cathode. This enabled
behavior of sulfur-oxidizing bacteria and heterotrophs in the a catholyte solution to be contained within the cathode
presence of a DC current. We established that, in the presence compartment and recirculated through an ion exchange
of soil, both Thiobacillus thiooxidans-like organisms and column for recovery of metals. The total volume of the
heterotrophs were protected from the otherwise deleterious catholyte system was approximately 500 mL with a recir-
effects of the electric current and indeed stimulation of culation rate of approximately 5 mL/min. A pH control unit
metabolic activity was observed (21). was incorporated to maintain an acidic pH (pH 4.5) in this
The purpose of the current study is to combine soil compartment through addition of acetic acid. An aqueous
acidification by indigenous sulfur-oxidizing bacteria with feed was attached to the anode, and electrical connections
electrokinetic remediation for removal of copper ions from were attached to a direct current power pack.
contaminated soil. A first series of experiments used silt soil contaminated
with 1000 mg/kg Cu(NO3)2 and adjusted to 30% (w/w)
Experimental Section moisture. The experiment shown here is a representative of
All studies were conducted using a standardized silt soil a series of three studies which were conducted under identical
obtained from Wye College, University of London (see Table conditions and yielded the same results. A current density
1 for characteristics). The soil was contaminated by spraying of 3.72 A/m2 electrode surface area was applied at a constant
a solution of metals onto the soil and mixing to effect a current with 8 M acetic acid added to the catholyte to
homogeneous contamination. For preliminary studies, a maintain an acidic pH, and 0.05 M H2SO4 was added to the
cocktail of six metals was used, added as nitrates, at anode at 60 mL/day.
concentrations of 10 mg/kg Cd, 500 mg/kg Cr, 500 mg/kg In a second set of experiments, the same soil was amended
Cu, 200 mg/kg Ni, 2000 mg/kg Pb, and 2000 mg/kg Zn, plus with 5% (w/w) sulfur and 30% (w/w) moisture and allowed
200 mg/kg molybdate. These concentrations were chosen to incubate for 90 days at 20 °C prior to electrokinetics. Two
such that the soil was characterized as contaminated electrokinetic experiments were conducted using this soil:
according to U.K. government guidelines (22). Following these one with 3.72 A/m2 current density and the other with the

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FIGURE 1. Electrokinetic apparatus. The power pack was able to deliver up to 2 A and 60 V. Acid reservoir contained 8 M acetic acid.
The pH control unit was set to add acetic acid when the pH in the catholyte reservoir, as measured by a pH probe, increased above pH
4.5. Recirculating pumps were used to maintain a flow of catholyte through the ion exchange column and the mixing reservoir at approximately
5 mL/min. Anolyte feed contained either 0.05 M sulfuric acid (first run) or water (second run) and was supplied at approximately 60 mL/day.
The box containing the anode and cathode was also filled with soil, and the movements of water and metal ions are shown.

The pH profile shows that in uncontaminated soil


TABLE 2. pH and Sulfate Contents of Metal-Contaminated and amended with 5% (w/w) sulfur acidification to approximately
Uncontaminated Soils Amended with 0.5 and 5% (w/w) Sulfur pH 1 could be achieved in 55 days. However, when this soil
(S) and Incubated at 20 ˚C at 30 % (w/w) Moisturea was contaminated with metals, the pH was only reduced to
21 days 42 days pH 2.5 over this time scale. Amendment of uncontaminated
sulfate sulfate
and contaminated soils with 0.5% (w/w) sulfur (results not
sample pH mg/kg soil pH mg/kg soil shown) led to acidification to pH 4 in both cases. Sulfate
generation above 2000 mg/kg was only seen with uncon-
0.5% S, uncontaminated 4.87 4910 4.31 8590 taminated soil amended with 5% (w/w) sulfur. In all other
0.5% S, contaminated 6.46 820 6.39 1200 flasks in which sulfur was added, sulfate generation reached
5% S, uncontaminated 5.01 5120 3.38 13770 1000-1500 mg/kg. This lower level of sulfate was sufficient
5% S, contaminated 6.28 1640 7.05 1320
to reduce the pH of the soil to between 2.5 and 4.5, but it was
control 7.78 40 7.74 0
not sufficient for further acidification. Separate analyses of
a The control soil was not amended with sulfur but was maintained
microbial populations by plating techniques indicated that
at constant (30% w/w) moisture for the duration of the experiment. The
pH of the soil at the start of the experiment was 8.1.
during the transition from pH 8 to pH 4, the predominant
species of sulfur-oxidizing bacteria present were neutrophilic
in terms of their growth pH (S. A. Jackman, unpublished
current density increased to 7.44 A/m2 after 7 days. In both results), whereas below pH 4, acidophilic bacteria, similar to
cases, 8 M acetic acid was added to the catholyte, and Thiobacillus thiooxidans, predominated (21). It has been
deionized water was added to the anode at 60 mL/day. demonstrated that at lower pH values metal ions such as
Metal was recovered using a column packed with ion- copper and zinc become soluble as ionic species in soil (23).
exchange resin (Amberlite IRC-718). The column was re- At this point, therefore, their toxicity to soil bacteria may
placed when copper adsorption became visible. Copper was increase. Nitrates and other anions are also more toxic to
eluted from the ion-exchange resin using a 15% (v/v) HCl sulfur oxidizing bacteria at low pH values, due to their
solution and analyzed by Atomic Absorption Spectroscopy protonation and potential movement into cells, destroying
(AAS). The total run times were 15 and 16 days for the first the membrane potential, ∆ψ (7). These toxic effects could
and second experiments, respectively. Soil metal concentra- cause the reduction in sulfur metabolism by indigenous
tions were determined by ED-XRF. sulfur-oxidizing bacteria under these conditions.
As a link to the electrokinetic experiments, which were
Results performed upon soil containing a single metal contaminant,
this copper-contaminated soil (1000 mg/kg copper nitrate)
Static Soil Experiment. Contaminated and uncontaminated was tested in a soil slurry (5% w/v) containing 10% (w/w)
silt soils were amended with sulfur at 30% (w/w) moisture sulfur. Similar profiles for pH and sulfate generation were
and incubated (Table 2). found as with the soil contaminated with a cocktail of seven
In the case of soil that had not been contaminated by metals.
addition of a cocktail of metals, sulfate was produced, and Electrokinetic Experiments. Standard silt soil contami-
the pH was reduced, whereas with metal-contaminated soil, nated to 1000 mg/kg with Cu(NO3)2 was subjected to
sulfate production was lower and the pH of the soil was not electrokinetics. No sulfur was added to the soil, and therefore
as much reduced. With 5% sulfur amendment, there was a no bacterial sulfur oxidation would be expected. The data in
10-fold difference in the level of sulfate production between terms of metal recovery at the cathode and overall pH of the
contaminated and uncontaminated samples, which was soil are displayed in Figure 3.
accompanied by a pH difference of approximately 3.7 units. Over 90% of copper ions were removed at the cathode
Soil Slurry Experiment. Silt soil was amended with sulfur within 15 days of initiation of the electrokinetic treatment.
to 5% (w/w) and made into a slurry at 30% (w/v). The results Similarly, there was removal of 70-90% of the calcium and
in terms of the change in pH and sulfate production for metal- manganese. Analyses of magnesium and iron were also
contaminated and uncontaminated soils are shown in Figure performed, but their movement was negligible under the
2. electric field. The soil temperature was between 25 and 35

VOL. 34, NO. 6, 2000 / ENVIRONMENTAL SCIENCE & TECHNOLOGY 9 1083


FIGURE 2. Acidification and sulfate production in silt soil slurries amended with sulfur. Uncontaminated silt soil was compared to soil
contaminated with a cocktail of seven metals. Data points for uncontaminated soil are shown by solid symbols and those for contaminated
soil by open symbols. Each flask (500 mL) contained 300 mL of 30% (w/v) soil slurry not amended with sulfur (diamonds) or amended with
5% w/w sulfur (squares). Solid lines are for pH values and dotted lines for sulfate concentration.

FIGURE 3. Metal recovery from the catholyte and the change in pH of the soil during electrokinetics. Recoveries are expressed as a
percentage of the starting metal concentrations in the soil. Copper ions are shown as triangles, calcium as squares, and manganese as
circles. pH is shown in diamonds with dotted lines.

°C. The operating voltage remained at an average of 12.2 V significantly below the buffering capacity for the soil,
for the first 9 days of the experiment, rising to an average of indicating that the acidic pH was as a result of the electro-
38.0 V for the remaining 6 days. The operating current was kinetics. Measurement of chloride and acetate concentrations
100 mA, and the average voltage over the whole experiment demonstrated migration of these ions toward the anode.
was 23.0 V. Soil moisture content was maintained at between Chloride concentration was reduced from 818 mg/kg to less
20 and 25% (w/w) over the course of the experiment. Water than 30 mg/kg across the soil within 24 h. Acetate concen-
balance determinations showed that 91% of water added to tration in the soil increased as the ion migrated into it from
the apparatus, in terms of soil moisture and liquid added to the catholyte. After 7 days, concentrations of approximately
the electrodes, was recovered at the end of the experiment. 10 000 mg/kg were determined across the whole area between
The pH and copper profiles across the soil, which was the anode and cathode.
divided into five centimeter-wide sections from anode to A second experiment was conducted, also with silt soil
cathode, were determined and are shown in Figure 4. contaminated with copper nitrate to 1000 mg/kg and with
The mass balance for copper was 100.1% as recovered the moisture adjusted to 30% (w/w), but, in this case, sulfur
within soil samples (assayed by ED-XRF) plus from the was added to 5% (w/w). In this experiment, the soil was
catholyte by ion exchange (assayed by AAS). Addition of preincubated for 90 days at 20 °C prior to the application of
sulfuric acid (approximately 86 mg/kg soil) at the anode was DC current, during which time the pH of the soil reduced

1084 9 ENVIRONMENTAL SCIENCE & TECHNOLOGY / VOL. 34, NO. 6, 2000


FIGURE 4. Copper concentration and pH profile across soil during electrokinetics of silt soil contaminated with 1000 mg/kg copper nitrate.
Individual time points are shown for time 0 (squares), 2 days (diamonds), and 15 days (circles). Solid lines are for copper concentration
and dotted lines for pH.

whereas iron and magnesium showed negligible movement.


Sulfate appeared to concentrate within the center of the
electrokinetic apparatus, but, as the electrokinetics pro-
gressed, it moved toward and into the anode compartment.
Initially, the soil contained 3101 mg/kg sulfate. After 10 days,
the level increased to an average of 15 939 mg/kg, an increase
of 5.1-fold. Within this same 10-day period, the average
copper concentration in the soil was reduced from 1000 mg/
kg to 186 mg/kg. After this time, the sulfate content of the
soil reduced to an average of 6417 mg/kg after 16 days as the
sulfate migrated into the anode compartment. Movements
and concentrations of chloride and acetate were similar to
those in the previous experiment.
An identical experiment in which the relative current
density was increased to 7.44 A/m2 after 7 days led to greater
FIGURE 5. Sulfate concentration across soil during electrokinetics. movement of sulfate ions toward the anode such that the
The soil was sectioned into five equal portions, the data for which
soil was cleared to less than 1000 mg/kg sulfate between the
are shown at 1-cm distances from the anode. Results (( SEM) are
electrodes.
displayed for time 0 (squares), 3 days (diamonds), 10 days (circles),
and 16 days (triangles). The power consumption for each of the electrokinetic
runs could be calculated from the applied voltage and current
from 8.1 to 5.4 due to the activity of the sulfur-oxidizing and the run time. For the first experiment the total electrode
bacteria. Average sulfate production was 3101 mg/kg soil. surface area was 0.0162 m2. With an operating current of 0.1
After the incubation period, the soil was packed into an A, this yields a current density of 6.2 A/m2. The average applied
electrokinetic reactor, and a DC current was applied under voltage was 23 V, over a distance of 6.5 cm between the
the same conditions as for the previous experiment with the electrodes. Assuming a linear increase in voltage at increasing
exception that water was added to the anode as opposed to distance between the electrodes, this gives a voltage of 354
sulfuric acid. Copper removed from the soil was collected by V/m. Power consumption was therefore 786 kWh/m3. At a
ion exchange, and soil samples were taken during the course power cost of $0.05/kWh and total run time of 15 days,
of the experiment. Analyses of the soil pH and copper assuming 1 m3 of soil is approximately 1.5 tonne, the total
concentration revealed very similar profiles to those displayed cost of metal removal was therefore $26.2 per tonne. When
in Figure 4, with pH reduced to below 3.0 in 9 days. The the soil was preincubated with sulfur-oxidizing bacteria,
average copper concentration across the soil was 114 mg/kg power consumption was 281 kWh/m3, yielding a cost of $9.3
after 16 days. Soil moisture content was maintained at per tonne, a cost reduction of 66%. The cost of sulfur, at
between 24 and 31% (w/w), and the soil temperature was below $250/tonne, and a relatively low input in terms of
between 20 and 27 °C over the course of the experiment. The man hours for monitoring and sampling do not greatly affect
applied voltage remained at an average of 6.3 V for the course this cost reduction.
of the experiment, apart from a peak of 28.2 V at day 8. Overall,
the average voltage was 7.7 V, with an operating current of Discussion
100 mA. Figure 5 shows the sulfate profile across the soil The potential of combining bioleaching by sulfur-oxidizing
during electrokinetics. bacteria with electrokinetics for metal mobilization and
The removal of copper from the soil at the cathode was recovery has been investigated. Initial experiments with sulfur
80% after 10 days and 86% after 16 days treatment. The final amendment of contaminated soil demonstrated that acidi-
mass balance for copper was 108%. Calcium and manganese fication was partially inhibited in comparison with uncon-
were also readily mobilized and removed from the soil, taminated soil in both static soil experiments and in soil

VOL. 34, NO. 6, 2000 / ENVIRONMENTAL SCIENCE & TECHNOLOGY 9 1085


slurries. Static experiments showed continuing production acidification of the soil below pH 4.5. This appeared to
of sulfate and reduction in pH in uncontaminated soil up to overcome the resistance which the sulfur-oxidizing bacteria
42 days. In comparison, after 21 days, there was no further encounter in attempting to reduce the pH of the soil to below
increase in sulfur oxidation in contaminated soil amended pH 3. The rise in sulfate production accompanied removal
with 5% sulfur. In soil slurries, enhanced sulfur oxidation of copper ions from the soil by electrokinetics. Experimental
and acidification might be expected due to increased aeration studies have also demonstrated that nitrate is rapidly removed
and dilution of the soil with water. While a reduction in pH (by electromigration toward the anode) from silt soil by
to below pH 3 was observed, in comparison to no significant electrokinetics (S. A. Jackman, unpublished results), although
pH change in static experiments, sulfate production remained it is also possible to speculate that microaerophilic sulfur
below 2000 mg/kg indicating that inhibition of bacterial oxidizers were able to reduce nitrate in the soil, thereby
activity was still occurring. Inhibition of bacterial metabolism relieving its inhibitory effects. It is possible that this removal
by metal ions has been studied for a wide range of bacteria of inhibitory metals and anions was sufficient for the activity
(23), and with acidophilic sulfur-oxidizing bacteria, copper of the sulfur-oxidizers to be stimulated. However, the rate of
and other ions have been shown to be inhibitory (24), sulfate production in the soil reached an average of 894 mg/
although the concentration of copper in this experiment may kg/day between days 3 and 7 and 1311 mg/kg/day between
not have been sufficient to produce the level of inhibition days 7 and 10. In comparison, sulfate production in un-
observed. Anions, especially nitrate, have also been shown contaminated soil in the static soil experiment was only 328
to be inhibitory at acidic pH values (7). Nitrate concentrations mg/kg/day. Clearly, the rate of sulfate production was
were significant in these experiments, and therefore this anion significantly increased in the presence of the electrokinetic
could be a significant contributor to the inhibition of bacterial treatment. It has been demonstrated previously that the
metabolism. The pH change from neutral to pH 4-5 was activity of sulfur-oxidizing bacteria is stimulated in soil slurries
accompanied by an alteration in the populations of indig- in the presence of an electric current (21). The soil used in
enous sulfur-oxidizing bacteria in the soil. Neutrophiles were these experiments was identical, and the microbial popula-
replaced by acidophiles. This pH change also affected the tions would be expected to be similar. The additional effects,
solubility of contaminating metal ions, with those such as which may be seen in soil treated with electrokinetics, include
copper and zinc becoming more soluble. The emergence of an increase in microbial activity due to a temperature rise
the acidophilic bacteria was therefore combined with the of between 3 and 13 °C. There is also the generation of oxygen
mobilization of inhibitory metal ions and the increased at the anode and the subsequent movement of oxygenated
toxicity of anions. The acidification process therefore slowed water into the soil. Since sulfur oxidation is very oxygen-
markedly. dependent, any increase in oxygen in the soil would be
Electrokinetic treatment of the soil generated protons at expected to positively affect this process. The stimulation of
the anode and hydroxyl ions at the cathode. While protons sulfur-oxidizing activity also accompanied an increase in
entered the soil and migrated toward the cathode by acetic acid (measured as acetate) movement from the
electromigration, hydroxyl ions were titrated at the cathode electrode and hence its concentration in the soil. A con-
with acetic acid to maintain the pH at 4.5 in this compartment. centration of 10 000 mg/kg corresponded to approximately
This process led to the soil being rapidly acidified to 670 mM acetate in the pore fluid of the soil. Experiments by
approximately pH 2 within a period of 9 days. At the same Alexander and co-workers (25) have demonstrated that, at
time, mobilized copper ions migrated toward the cathode a concentration of 10 mM (pH 3), acetic acid accumulates
by electromigration and entered the cathode chamber where in the cytoplasm of T. ferrooxidans and partially inhibits the
they were pumped through an ion exchange column. oxidation of ferrous iron. The findings of the present study
Removal of copper was 85% complete within 15 days when in which the soil contained much higher concentrations of
soil had not been preacidified due to the activity of sulfur- acetic acid clearly demonstrate that these were not inhibitory
oxidizing bacteria. For preacidification experiments, a time to sulfur oxidation by sulfur-oxidizing bacteria. An additional
scale of 90 days was chosen for incubation with sulfur, to experiment, in which a higher current density was used
maximize sulfate production and acidification based upon resulted in removal of sulfate from the soil to below 1000
the results of earlier experiments. The observation that, in mg/kg, is of significance. One of the major drawbacks of
static experiments, bacterial activity had ceased after 21 days bioleaching technology is the high production of sulfate from
in the presence of seven contaminating metals, whereas in sulfur/sulfide leading to soil contaminated with sulfate. The
uncontaminated soil, activity continued up to 42 days (and selective and efficient removal of sulfate from soil under
possibly beyond) meant that in studies with a single electrokinetic processing following metal removal may
contaminating metal, a time scale of 90 days might be therefore lead to a cleaner and more effective technology.
expected to ensure maximal sulfate generation and acidi- In summary, the combination of preacidification by sulfur-
fication. When the soil was allowed to preacidify, there were oxidizing bacteria followed by electrokinetics is cost-effective
two marked differences in the overall run parameters. First, in reducing the power input for electrokinetics. The elec-
copper removal was achieved with much lower power trokinetic treatment also appears to stimulate the activity of
consumption. The pH of the soil had already been reduced sulfur-oxidizing bacteria by removal of inhibitory ions and
by 2 units due to sulfur oxidation, and therefore the amount other positive effects of the electric current upon soil
of hydrogen ions required to acidify the soil to pH 2 and the microbial activity. These synergistic effects are promising
power required to generate these ions were reduced. In
for future experiments in which complex mixtures of metal
commercial application in a field situation, the cost of power
ions may be present, and bacteria may be required to mobilize
is a major component of the overall cost of the process. In
metals from sulfides. The methodology has potential for a
addition, any reduction in the time scale for electrokinetic
range of contaminated sites including former gasworks and
remediation will also affect the cost in terms of manpower
wastes from mining.
and requirements for equipment. By amending soil at an
earlier stage with little further intervention, the soil can be
prepared for electrokinetic treatment. Acknowledgments
A second benefit of the preacidification experiment is This work was supported by the U.K. Department of Trade
that sulfate production by sulfur-oxidizing bacteria was and Industry and the Engineering and Physical Sciences
significantly stimulated when the electric current was applied. Research Council under the LINK Biological Treatment of
One of the effects of the electrokinetics was to enable Soils and Water Initiative.

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