Sunteți pe pagina 1din 10

Environ Geochem Health

DOI 10.1007/s10653-007-9095-z


The effects of Acidithiobacillus ferrooxidans on the leaching

of cobalt and strontium adsorbed onto soil particles
Hyun-Sung Park Æ Jong-Un Lee Æ Jae-Woo Ahn

Ó Springer Science+Business Media B.V. 2007

Abstract Bioleaching from soil artificially contam- the same leaching conditions, the extent of Sr
inated with analogues of radionuclides, Co and Sr, removal was much lower than that of Co. On the
was carried out using a Fe-oxidizing bacterium, contrary to the high efficiency of microbial metal
Acidithiobacillus ferrooxidans. Due to bacterial leaching in biohydrometallurgy for low-graded sul-
metabolism, the pH and dissolved Fe3+ concentration fide ores, which has been widely documented,
in a biotic slurry decreased and increased respec- conventional bioleaching techniques with A. ferroox-
tively, over time, but the concentrations of Co and Sr idans supplied with enough Fe2+ showed low effi-
extracted from the soil showed no significant ciency for the removal of radionuclides loosely bound
enhancement compared with those under abiotic onto soil particle surfaces.
control. In both cases, Co and Sr were leached from
the soil during the initial period of the experiment,
due to the initially low solution pH of 2.0, and the Keywords Bioleaching  Radionuclides  Cobalt 
dissolved concentrations remained almost constant Strontium  Acidithiobacillus ferrooxidans  Soil
for the duration of the experiment (300 h). Since
oxidation of Fe2+ by A. ferrooxidans led to the
production of Fe precipitates and colloidal suspen- Introduction
sions, the Co and Sr extracted into solution were most
likely re-adsorbed onto the Fe solids. Also, A. Contamination of soil and sediment with radionuc-
ferrooxidans, without an external supply of Fe2+, lides can pose significant hazards to human health. In
extracted almost equal or greater amounts of Co and particular, unexpected accidents at nuclear power
Sr from the soil than when Fe2+ was supplied. Under plants, e.g., in Chernobyl, and potential release of
radioactive elements from disposal sites, will signif-
icantly exert malign influences on the biosphere.
When surface environments are polluted with radio-
H.-S. Park  J.-U. Lee (&)
nuclides, the application of natural containment
Microbial Geochemistry Lab, Department of Geosystem
Engineering, Chonnam National University, Gwangju technology, through sorption or precipitation, will
500-757, Korea never be a radical remediation method due to the very
e-mail: long half-lives of most radionuclides. Therefore, once
soil and sediment pollution with radioactive elements
J.-W. Ahn
Department of Advanced Materials Science and has occurred, thorough clean-up processes should be
Engineering, Daejin University, Pocheon 487-711, Korea performed immediately.

Environ Geochem Health

Chemical and bacterial leaching methods have (Schippers et al. 1996; Fowler and Crundwell 1998),
been developed for the decontamination of soil and the bacterial production of Fe3+ and subsequent sulfide
sediment polluted with heavy metals. Chemical dissolution play a dominant role in bioleaching
treatment, with various oxidizing, reducing, and processes.
complexing agents, has been used for the extraction
bacteria þ
of heavy metals from contaminated soil, sediment, MeS2 + H2 O + 3.5 O2 ! Me2þ + 2 SO2
4 + 2H
and sewage sludge (e.g., Kedziorek et al. 1998; Steele ð3Þ
and Pichtel 1998; Wasay et al. 1998). However, the
high cost, operational difficulties, intensive energy
The processes described above (Eqs. 1–3) require
requirements, unsatisfactory yield of metal solubili-
sulfides to be oxidized, with the production of
zation and hazardous byproducts have caused
hydrogen ions, which finally lowers the system pH.
researchers to consider alternative methods (Tyagi
However, natural soil, sediment, and sludge contam-
and Couillard 1989). The bioleaching process has
inated with heavy metals generally contain only small
been proposed as an effective, economic, and envi-
amounts of sulfur and sulfides, with the exception of
ronmentally-friendly method for the treatment of
those related to mining activity. In these cases,
heavy metals and radionuclides.
however, the acidification of non-sulfide solids by
One of the most widely used microorganisms in
A. ferrooxidans can also be performed by Fe2+
bioleaching is the acidophilic Acidithiobacillus fer-
oxidation (Eq. 4), followed by the precipitation of
rooxidans (Blais et al. 1992). This lithoautotrophic
ferric iron in the form of jarosite (Eq. 5) or ferric
bacterium gains energy to support growth and
hydroxide (Gu and Wong 2004).
maintenance from the oxidation of Fe2+ or reduced
sulfur compounds, and uses CO2 as a cellular carbon bacteria
source. Therefore, an additional carbon source or 4 Fe2þ + O2 + 4 Hþ ! 4 Fe3þ + 2H2 O ð4Þ
electron donor is not required. When A. ferrooxidans
oxidizes Fe2+ to Fe3+, it produces protons and an
oxidant (Fe3+), which are critical factors in the 3 Fe3þ + Meþ + 2 HSO
4 + 6 H2 O
leaching process, especially of sulfides. ! MeFe3 ðSO4 Þ2 ðOHÞ6 + 8 Hþ
Acidithiobacillus ferrooxidans oxidizes ferrous to
ferric ions (Eq. 1), with the latter subsequently (Me+: free metal ion)
leading to the oxidative dissolution of sulfides (Eq. Acidithiobacillus ferrooxidans has been practically
2). Overall reactions produce extra hydrogen ions used in the hydrometallurgy of low-graded sulfide
and, thus, the system pH is lowered (Boon et al. 1998; ores since Fe3+ can lead to the oxidative dissolution
Fowler and Crundwell 1998). of sulfides (e.g., Dew et al. 1998; Dastidar et al.
2000). Moreover, much recent research has indicated
bacteria that the bioleaching technique can be successfully
14 Fe2þ + 3.5 O2 + 14 Hþ ! 14 Fe3þ + 7H2 O
applied to solving environmental problems related to
ð1Þ heavy metals. They reported many good results on
the effective biolixiviation of heavy metals from
þ contaminated soil, sediment, and sewage sludge using
MeS2 +8H2 O+14Fe3þ !Me2þ +14Fe2þ +2SO2
4 +16H Fe-oxidizing bacteria (e.g., Sreekrishnan et al. 1993;
ð2Þ Gomez and Bosecker 1999; Chen and Lin 2000; Jang
et al. 2002; Kim et al. 2005). The efficiency of the
(MeS2: insoluble metal sulfides, Me2+: free metal microbial leaching process is affected by various
ion) parameters, such as the bacterial strain used, the
Some researchers have suggested that the direct quantity and type of substrate, and the solid content,
contact of bacteria with sulfide surfaces results in particle size, and carbon dioxide and oxygen con-
metal dissolution (Eq. 3; Silverman and Ehrlich 1964; centrations in the system, as well as the pH and redox
Free et al. 1991; Lizama and Suzuki 1991). However, potential (Couillard and Zhu 1992; Haddadin et al.
according to some recent well-designed investigations 1995).

Environ Geochem Health

Despite extensive research on heavy metals biole- (Sigma-Aldrich) and 88Sr(NO3)2 (Sigma-Aldrich)
aching, few studies have been performed on the respectively were used to contaminate the clean soil.
remediation of soil and sediment contaminated with After pollution, the soil sample was washed three
radionuclides. Since the released radioactive ele- times with distilled, de-ionized water and dried at
ments will pollute soil and sediment, mainly by 358C. The concentrations of Co, Sr, and Fe in the
adsorption via air and natural waters, they will exist original, uncontaminated and the artificially contam-
in a loosely bound mode in geological media. Thus, inated soils were determined after extraction by aqua
the contaminants in radioactively polluted soil and regia (HCl:HNO3 = 3:1) for 1 h at 708C .
sediment differ in their mode of occurrence from that
of sulfide ore minerals, where the metals reside in a Bacteria and culture conditions
strong crystal lattice structure. Therefore, whether Fe-
oxidizing bacteria can leach the metals or radionuc- A freeze-dried culture of A. ferrooxidans (KCTC
lides adsorbed onto solid surfaces, with a high 2677) was obtained from the Korean Collection of
leaching efficiency, remains to be determined. Type Culture. Cells were cultivated in 100 ml of 9 K
In order for A. ferrooxidans to maintain active medium in a 500-ml flask (Silverman and Lundgren
metabolism during the bioleaching process, an exter- 1959) placed in a shaking incubator at 308C and
nal supply of both Fe2+ and inorganic nutrients, such 200 rpm. The 9 K medium consisted of two types of
as NH+4 , K+, HPO2 2+ 2+
4 , Mg , and Ca , is required. solutions; solution A: 3 g (NH4)2  SO4, 0.1 g KCl,
Recently, Lee et al. (2005) reported that a supply of 0.5 g K2HPO4, 0.5 g MgSO4  7H2O, and 0.014 g
inorganic nutrients from natural soil for A. ferroox- Ca(NO3)2  4H2O in 700 ml of distilled water, and
idans led to a slightly higher leaching efficiency of solution B: 45 g FeSO4  7H2O in 300 ml of distilled
uranium than with the artificial, external addition of water. Solutions A and B provided inorganic nutri-
nutrients. Since the cost of supplying nutrients to the ents and an energy source to the bacterial cells
bacterial cells can be reduced, if this were the case, respectively. Serial transfer was performed several
the experimental determination of the optimal leach- times to enhance the cell activity.
ing conditions will be required.
The objectives of this study were to examine the Leaching experiments with and without bacteria
bioleaching capacity of A. ferrooxidans for the
extraction of radionuclides adsorbed onto the surfaces Batch-type bioleaching and abiotic control leaching
of soil particles, and provide fundamental informa- experiments were conducted according to the fol-
tion on the highly efficient and economic conditions lowing conditions:
required for the leaching process.
– Experimental set (1): Soil (25 g) + solution A
(350 ml) + solution B (150 ml) + A. ferrooxidans
(50 ml)
Materials and methods
– Experimental set (2): Soil (25 g) + solution A
(350 ml) + solution B (150 ml) + sterile distilled
Artificial contamination of soil samples
water (50 ml)
Uncontaminated soil samples were taken in the The experiments were performed in 1,000-ml
vicinity of Yeonggwang nuclear power plant, which conical flasks, each containing 25 g of artificially
is located in the southwestern part of Korea. The contaminated soil and 500 ml of 9 K medium, in a
soil was dried at room temperature, and the physi- shaking incubator at 308C and 200 rpm for 300 h.
cochemical properties of soil (cation exchange Solution A of the 9 K medium was autoclaved, and
capacity, soil pH, loss-on-ignition, and textural solution B was sterilized by passing through a 0.2-mm
analysis) determined. The soil sample (<0.18 mm filter. The bioleaching flasks were inoculated with
diameter) was soaked in a mixed solution of Co and 50 ml of A. ferrooxidans, which had been cultured for
Sr (each 1,000 mg/l) for 24 h. Instead of radioactive 30 h. The pH of the solution was adjusted to 2.0 with
Co (half-life = 5.27 years) and 90 Sr (half- 5 M H2SO4 and 0.1 M NaOH. Periodically, aliquots
life = 28.8 years), non-radioactive 59CoCl2  6H2O were taken from each flask, filtered through an

Environ Geochem Health

0.2-mm filter and the pH as well as dissolved Fe2+, modified ferrozine method (Stookey 1970). For this, a
total Fe, Co, and Sr concentrations of the solution 0.1-ml sample was rapidly mixed with 4.9 ml of 0.1%
were determined. ferrozine solution in 0.5% pH 7.0 ammonium acetate
buffer (adjusted with NaOH). The solution was
Bioleaching experiments under various conditions agitated, and then assayed using a UV-vis spectro-
photometer (UV-MINI-1240) at 562 nm. The con-
To determine the experimental conditions leading to centration of ferric ion was calculated by subtracting
high bioleaching efficiency, the experimental sets the measured Fe2+ concentration from that of the total
were designed, as shown below, with the leaching dissolved Fe.
capacities then compared: The total Fe, Co, and Sr concentrations in the
slurries and residual soils were determined using
– Experimental set A: Soil (25 g) + solution A
ICP-AES (Jobin Ybon JY70+).
(350 ml) + solution B (150 ml) + A. ferrooxidans
(50 ml)
– Experimental set B: Soil (25 g) + distilled water
Results and discussion
(350 ml) + solution B (150 ml) + A. ferrooxidans
(50 ml)
Characteristics of the soil sample
– Experimental set C: Soil (25 g) + solution A
(350 ml) + distilled water (150 ml) + A. ferro-
The studied soil was slightly acidic (pH = 5.0), and
oxidans (50 ml)
had a silty loam texture (sand 2.3%, silt 64.9%, clay
– Experimental set D: Soil (25 g) + distilled water
32.8%). The cation exchange capacity was 30.0 meq/
(500 ml) + A. ferrooxidans (50 ml)
100 g, with loss-on-ignition of 4.5%. The concentra-
Experimental set (A) was designed to examine the tions of Co and Sr in the uncontaminated soil were 30
bioleaching under 9 K medium conditions, which are and 21 mg/kg respectively. The artificially contam-
those most widely employed for A. ferrooxidans inated soil had 373 and 393 mg/kg of Co and Sr
activity. In the case of set B, solution A (the inorganic respectively. After soaking for only 24 h, the levels of
nutrients for cellular growth and synthesis) was not Co and Sr contamination in the soil samples were 10
supplied to the slurry. In experimental set C, solution times those of the uncontaminated soil. The concen-
B (Fe2+, which acts as an energy source for growth tration of Fe in the soil was 1.60%.
and maintenance of A. ferrooxidans), was absent. In
experimental set D, neither solution A nor B was Leaching efficiency of A. ferrooxidans and abiotic
added. A. ferrooxidans was infused to the system control
after 30 h of incubation in the 9 K medium. The pH
of the solution was adjusted to 2.0 with 5 M H2SO4 Batch-type leaching experiments were conducted
and 0.1 M NaOH. After the periodic removal of under A. ferrooxidans (the set 1) and abiotic control
aliquots from the batch-type slurries, the solution pH (the set 2) conditions. The bacteria provided with
and dissolved Fe2+, total Fe, Co, and Sr concentra- solutions A and B of the 9 K medium as external
tions were measured after filtration through a 0.2-mm nutrient and energy sources produced a greater
filter. amount of protons, which resulted in a lower solution
After 300 h, the residual soils in each flask were pH than that in the abiotic control (Fig. 1). This result
removed, dried at room temperature, and the Co, Sr, indicated that the iron oxidizer included in set 1
and total Fe then extracted by aqua regia digestion performed normal metabolism, where A. ferrooxidans
and determined. oxidized Fe2+ to Fe3+, with H+ supplied into the
solution by the chemical formation of Fe precipitates
Analytical techniques (Eqs. 4, 5); however, the abiotic control showed a
gradual increase in the solution pH with time, as
The pH was measured using a glass electrode hydrogen ions were chemically consumed during the
connected to an Orion Model 720A pH meter. The protonation and dissolution of solid particles. Since
concentration of ferrous ion was determined by a the pH is apparently one of the factors significantly

Environ Geochem Health

2.6 10

Dissolved Fe concentrations (g/L)

8 dissolved
2.4 3+

2 3+
dissolved Fe
1.8 Set (1)
Set (2) 0
0 50 100 150 200 250 300
0 50 100 150 200 250 300 Time (hours)
Times (hours) Fe(T) in set (1) Fe2+ in set (1)
Fe(T) in set (2) Fe2+ in set (2)
Fig. 1 Variation in pH values in the leaching experiments with
A. ferrooxidans and the abiotic control over time. Set 1, Fig. 2 Variations in aqueous total Fe (Fe(T)) and Fe2+
microbial leaching with A. ferrooxidans; set 2, abiotic control concentrations in the leaching experiments with A. ferrooxi-
dans over time
mediating the rate and extent of metal leaching, the
decrease in the pH due to A. ferrooxidans would processes has been extensively documented by many
imply that the use of this bacterium can lead to researchers (e.g., Tuovinen 2004). However, the
favorable conditions for the leaching of metals from abiotic control showed no significant decrease in
various solids, and this strongly acidic environment the total Fe concentration, and by the end of the
will be maintained over a relatively long period. experiment, more than 85% of the dissolved Fe
The amount of Fe2+, used as the energy source in existed as Fe2+.
the 9 K medium in these experiments, was 9 g/l. At a The amounts of Co and Sr extracted showed no
given time during the leaching process, the difference remarkable differences between the biotic samples
between input Fe (9 g/l) and total dissolved Fe and abiotic controls (Fig. 3). Moreover, the concen-
contents are an indication of the amounts of Fe trations of Co and Sr in experimental set 1 were lower
precipitates and colloidal suspensions. Also, the than in the abiotic controls for the duration of the
difference between total dissolved Fe and Fe2+ will experiment.
give the amount of dissolved Fe3+. Aqueous Fe3+ This result was most likely due to the mode of
serves as a strong oxidant, which greatly facilitates occurrence of Co and Sr in the soil, as well as the
metal extraction from solid substrates, especially mineralogical type of solid to be leached. In the
sulfides (Eq. 2). The production of Fe precipitates, experiment, the soil was soaked in the Co and Sr
such as jarosite, can also lower the solution pH, thus solution for 24 h to simulate radionuclide pollution
enhancing metal leaching (Eq. 5). Therefore, the and, thus, that the Co and Sr might become loosely
microbial production of aqueous Fe3+ in solution is bound onto the soil surface through adsorption. With
essential for an effective leaching process. regard to the conditions, an initial pH of 2 was low
The A. ferrooxidans in experimental set 1 oxidized enough to extract the observed amounts of Co and Sr
nearly all the Fe2+ and produced approximately 4 g/l in the abiotic control. This can be confirmed by the
of Fe3+ within 110 h (Fig. 2). The concentration of amounts of dissolved Co and Sr abruptly extracted
total dissolved Fe also decreased over time in set 1. during the initial period of experiments.
The reduced amount of total dissolved Fe (about 5 g/l If the Co and Sr are associated with sulfide
in set 1) was considered to have formed Fe precip- minerals, the low pH and large amount of dissolved
itates and colloidal suspensions. During experiment Fe3+ in set 1, which were the characteristic results of
1, the formation of reddish brown precipitates and the microbial leaching process (Figs. 1, 2), might
colloidal suspensions was readily observed in the significantly enhance the rate and extent of leaching
flask employed. The transformation of Fe2+ into a over time.
variety of Fe minerals, such as jarosite, schwertman- The formation of Fe precipitates and colloidal
nite, goethite, and ferrihydrite, during bioleaching suspensions following the oxidation of Fe2+ to Fe3+

Environ Geochem Health
Dissolved Co concentrations (mg/L)
30 The bioleaching process using Fe-oxidizing bac-
teria has been proved to be one of the most promising
methods for the extraction of heavy metals from
20 contaminated soil and sediment. However, the pres-
ent study has indicated that such elevated bioleaching
efficiency can be constrained by the mode of
10 occurrence of the heavy metals, as well as the
5 Set (1) mineralogical characteristics of soil and sediment.
Set (2)
0 50 100 150 200 250 300 Bioleaching under various operational conditions
Time (hours)
Batch-type bioleaching was performed using exper-
Dissolved Sr concentrations (mg/L)

imental sets A, B, C, and D, as described above. The
25 bacteria provided with solution B (Fe2+) of the 9 K
medium as an energy source (experimental sets A and
B) produced the high amounts of protons, resulting in
15 lower solution pHs than in experimental sets C and D,
where no Fe2+ was externally supplied (Fig. 4).
Set (1)
Although the pH in experimental set B was
5 slightly higher than that in set A, which was also
Set (2)
provided with inorganic nutrients, a dramatic pH
0 50 100 150 200 250 300 decrease was observed over time. Considering that
Time (hours) metabolically active A. ferrooxidans continuously
produces protons, the bacterium, when not supplied
Fig. 3 Variations in the dissolved Co and Sr concentrations in
the leaching experiments with A. ferrooxidans with inorganic nutrients, also appeared to perform
active metabolism. As described earlier, this result
was probably due to the natural supply of a variety of
may explain why experimental set 1 showed slightly inorganic nutrients to the bacteria from the soil
lower concentrations of dissolved Co and Sr than the substrate. The elements leached from the soil due to
abiotic control. The Fe precipitates and colloidal the initially low pH might provide enough inorganic
suspensions probably adsorbed the dissolved Co and
Sr in the solution and, thus, mediated the aqueous
concentrations of these metals. In experimental set 2, 2.6
where no bacteria were infused, the solution was
observed to be less stained than in set 1, indicating 2.4

the formation of many fewer Fe precipitates and

colloidal suspensions. The chemical oxidation of Fe2+

without microbial reactions is known to be slower

than that of A. ferrooxidans by 105–106 times (Lacey Set (A)
and Lawson 1970). Set (B)
Set (C)
The fact that more Co than Sr was extracted both
Set (D)
microbially and chemically, as shown in Fig. 3, 1.6
0 50 100 150 200 250 300
indicates that the extraction efficiency may depend on
Time (hours)
the type of radionuclides. Conceivably, Sr may be
more readily adsorbed onto other solids or formed Fig. 4 Variation in pH values in the leaching experiments with
precipitates or colloidal suspensions than Co. Since A. ferrooxidans under various conditions over time. Set A,
this alkaline earth element, which easily forms stable bioleaching of A. ferrooxidans with inorganic nutrients and
Fe2+; set B, bioleaching of A. ferrooxidans with Fe2+; set C,
sulfate precipitate, has not been the focus of environ- bioleaching of A. ferrooxidans with inorganic nutrients; set D,
mental investigations, the result is relatively new. bioleaching of A. ferrooxidans with water

Environ Geochem Health

nutrients to support the normal metabolism of A. where no inorganic nutrients were initially provided,
ferrooxidans. This result is of economic significance, there was a higher production of dissolved Fe3+ than
as the expense for inorganic nutrients externally in set A. Approximately 2.4 g/l of dissolved Fe3+ was
supplied to the bacterium can be reduced. additionally present in set B after 150 h compared
On the contrary, in experimental sets C and D, with set A. No initial supply of inorganic nutrients
little variation in the pH was observed over time once resulted in the inhibition of Fe precipitates, and, thus,
the pH had leveled out after the beginning of the a higher amount of dissolved Fe3+ in solution.
experiment. Ferrous ions, the essential energy source The amounts of Co extracted in sets A and B were
for A. ferrooxidans, was not supplied to either set; slightly greater than those in sets C and D, where no
therefore, unlike inorganic nutrients, a sufficient Fe2+ was supplied, although the difference dimin-
amount of Fe2+ to support bacterial metabolism ished after 250 h (Fig. 6). In comparison, the Sr
seemed to be naturally depleted in the studied soil. concentrations showed no significant difference
The iron speciation over time, i.e., dissolved total between the four batch-type slurries studied. The
Fe and Fe2+, in experimental sets A and B is depicted amounts of Sr extracted were almost the same in all
in Fig. 5. The amounts of Fe that formed precipitates sets, and appeared to be unaffected by the various
and colloidal suspensions in sets A and B were experimental conditions.
calculated to be 5.3 and 2.9 g/l respectively, after The extractions of Co and Sr were also signif-
150 h. According to Eq. 5, which indicates that Fe icantly influenced by the initial pH. The Co and Sr in
precipitation leads to a pH decrease, the result the artificially contaminated soil might be adsorbed
corresponded with the difference in solution pHs onto the surface of soil particles, and more easily
shown in Fig. 4. Interestingly, in experimental set B, dissolved by the low initial pH. Furthermore, the

Dissolved Co concentrations (mg/L)

Set (A) Total Fe

Dissolved Fe concentrations (g/L)

8 Fe2+ 25

6 Fe in precipitates and
colloidal suspensions 15
4 Set (A)
Set (B)
2 5 Set (C)
Dissolved Fe Set (D)
0 0 50 100 150 200 250 300
0 50 100 150 200 250 300
Time (hours)
Time (hours)
Dissolved Sr concentrations (mg/L)

Set (B) Total Fe
Dissolved Fe concentrations (g/L)

Fe in precipitates and 20
colloidal suspensions
6 15

Set (A)
4 Set (B)
3+ 5 Set (C)
Dissolved Fe
2 Set (D)
0 50 100 150 200 250 300
0 50 100 150 200 250 300
Time (hours)
Time (hours)
Fig. 6 Variations in the dissolved Co and Sr concentrations in
Fig. 5 Variations in the aqueous total Fe and Fe2+ concentra- the leaching experiments with A. ferrooxidans under various
tions for bioleaching under various conditions over time conditions over time

Environ Geochem Health

formation of Fe precipitates and colloidal suspensions 100

Co Sr 90 91

Percentage of removal (%)

probably acted as strong adsorbents, and, thus, there 77
were no increases in the amounts of dissolved Co and 72
Sr in the slurries over time. 60
The amounts of Fe colloidal suspensions in the
final solution after the 300 h of the experiment were 40
32 36
approximately calculated for the four slurries, accord-
ing to Fe mass balance equation, as follows; Fe in 20

colloidal suspensions = (Fe in the original soil + Fe in

the bacterial inoculum + Fe in the solution B of 9 K A B C D
medium)  (Fe in the residual soil after bioleach- Experimental sets
ing + dissolved Fe in the final solution). When Fe2+
Fig. 8 The percentage Co and Sr removals under various
was supplied to A. ferrooxidans, the amounts of Fe
experimental conditions. The numbers near each bar represent
colloidal suspensions were higher than those of Fe the percentage removal
precipitates (Fig. 7). In the practical application of a
bioleaching technique for the removal of radionuc-
lides from contaminated soil (e.g., soil washing), the precipitates or colloidal suspensions were formed;
colloidal suspensions probably adsorb the released therefore, no re-adsorption of extracted Co and Sr
radionuclides, which can be easily separated from the onto solid Fe compounds could take place. In
residual soil, and then treated similarly to dissolved particular, Sr showed a much lower extent of removal
radionuclides. when Fe2+ was externally supplied. While A. ferro-
More Co than Sr was extracted. The difference oxidans supplied with Fe2+ extracted only 32–36% of
between the two elements was also reflected in the the Sr, the bacterium with no Fe2+ initially supplied
analytical results obtained for the residual soils after removed 65–77% of the Sr from the soil. Strontium
the bioleaching process. The residual soils were may be adsorbed more by Fe solids or precipitated
digested with aqua regia, and the Co and Sr after the formation of compounds associated with
concentrations determined. Under a range of given dissolved anions, such as sulfate.
experimental conditions, Sr remained in the residual This result suggested that preliminary investiga-
soils at higher concentrations than Co (Fig. 8). tions are required on the type of contaminant
It is remarkable, according to the final percentages radionuclides and the chemical and mineralogical
of Co and Sr removals from the contaminated soil, composition of the contaminated soil to be leached
that experimental sets A and B, with Fe2+ added, for a cost-effective treatment to be implemented.
showed almost equal or lower removals than sets C Since the reduced form of Fe (Fe2+) is essential for
and D. Because sets C and D did not contain large the normal metabolism of A. ferrooxidans, sufficient
amounts of initially supplied Fe2+, no significant Fe Fe2+ should be externally provided to the bacterium
to achieve satisfactory results from microbial metal
5000 extraction. To date, a number of results, with high
Fe in colloid
Fe in residual soil efficiency of microbial metal lixiviation, have been
documented for low-graded sulfide ore minerals, and
Fe mass (mg)

3000 even for soil and sediment contaminated with heavy

metals. However, according to the results presented
2000 in this study, such good leaching efficiency may not
be expected when the contaminants are present in an
adsorbed mode in the soil or sediment substrate; e.g.,
0 radionuclide pollution through fall-out. Furthermore,
A B C D in such cases, the initial supply of Fe2+ to stimulate
Experimental sets
the Fe-oxidizer will form Fe solids, which re-adsorb
Fig. 7 Iron mass in colloidal suspensions and residual soil; the metals leached into the solution. Only low pH
calculated using a mass balance equation conditions may be adequate to efficiently extract such

Environ Geochem Health

weakly bound metals. Further investigations will be Chen, S.-Y., & Lin, J.-G. (2000). Influence of solid content on
required to examine the metal leaching efficiency of bioleaching of heavy metals from contaminated sediment
by Thiobacillus spp. Journal of Chemical Technology and
leaching agents with a low pH and low Fe3+ Biotechnology, 75, 649–656.
concentrations, which can be easily achieved by the Couillard, D., & Zhu, S. (1992). Bacterial leaching of heavy
simple addition of a weak acid. metals from sewage sludge for agricultural application.
Water, Air, and Soil Pollution, 63, 67–80.
Dastidar, M. G., Malik, A., & Roychoudhury, P. K. (2000).
Biodesulphurization of Indian (Assam) coal using Thio-
Conclusion bacillus ferrooxidans (ATCC 13984). Energy Conversion
and Management, 41, 375–388.
Bioleaching and abiotic leaching of soil contaminated Dew, D. W., Lawson, E. N., & Broadhurst, J. L. (1998). The
BIOX1 process for biooxidation of gold-bearing ore or
with Co and Sr were conducted. In all cases, the concentrates. In D. E. Rawlings (Ed.) Biomining: Theory,
removal of Co and Sr from the soil was facilitated at Microbes and Industrial Processes. (pp. 45–80). Berlin:
an initially low pH, as Co and Sr were present in a Springer.
loosely bound mode, via adsorption, onto the soil Fowler, T. A., & Crundwell, F. K. (1998). Leaching of zinc
sulfide by Thiobacillus ferrooxidans: Experiments with a
surface. The extent of Sr removal was lower than that controlled redox potential indicate no direct bacterial
of Co. A. ferrooxidans showed no significant mechanism. Applied and Environmental Microbiology,
enhancement for the extractions of Co and Sr 64, 3570–3575.
compared with the abiotic control, although the Free, M. L., Oolman, T., Nagpal, S., & Dahlstroom, D. A.
(1991). Bioleaching of sulphide ores distinguishing
bacterium produced many protons and much dis- between indirect and direct mechanisms. In R. W. Smith
solved Fe3+. Due to re-adsorption onto the Fe & M. Misra (Eds.), Mineral Bioprocessing. (pp. 485–495).
precipitates and colloidal suspensions readily formed Warrendale: The Minerals, Metals and Materials Society.
by A. ferrooxidans in the case of Fe2+ addition, the Gomez, C., & Bosecker, K. (1999). Leaching heavy metals
from contaminated soil by using Thiobacillus ferrooxi-
amounts of dissolved metals were almost equal to or dans or Thiobacillus thiooxidans. Geomicrobiology
lower than those when no Fe2+ was supplied. Journal, 16, 233–244.
The conventional bioleaching technique, using Fe Gu, X., & Wong, J. W. C. (2004). Identification of inhibitory
oxidizers, e.g., A. ferrooxidans, may show no enhanced substances affecting bioleaching of heavy metals from
anaerobically digested sewage sludge. Environmental
efficiency for removal of radionuclides from natural Science and Technology, 38, 2934–2939.
soils when supplied with sufficient Fe2+. This result Haddadin, J., Dagot, C., & Fick, M. (1995). Models of bacterial
indicated that preliminary investigations are required leaching. Enzyme and Microbial Technology, 17, 290–305.
on the chemical and mineralogical compositions of the Jang, A., Jang, H.-Y., Kim, S.-M., Lee, J.-U., & Kim, I. S.
(2002). Decontamination of heavy metals from dewatered
contaminated soil to be leached, as well as the mode of sludge by Acidithiobacillus ferrooxidans. Environmental
occurrence of radionuclides or heavy metals, for a Engineering Research, 7, 199–206.
cost-effective treatment to be implemented. Kedziorek, M. A. M., Dupuy, A., Bourg, A. C. M., & Compère,
F. (1998). Leaching of Cd and Pb from a polluted soil
Acknowledgement We appreciate the critical comments of during the percolation of EDTA: Laboratory column
an anonymous reviewer, which greatly improved this experiments modeled with a non-equilibrium solubiliza-
manuscript. This work has been financially supported by a tion step. Environmental Science and Technology, 32,
grant from the KESRI (Korea Electrical Engineering and 1609–1614.
Science Research Institute; R-2003-B-351) to J.-U. Lee, which Kim, I. S., Lee, J. -U., & Jang, A. (2005). Bioleaching of heavy
was funded by the MOCIE (Ministry of Commerce, Industry metals from dewatered sludge by Acidithiobacillus
and Energy), Korea. ferrooxidans. Journal of Chemical Technology and
Biotechnology, 80, 1339–1348.
Lacey, D. T., & Lawson, F. (1970). Kinetics of the liquid-phase
oxidation of acid ferrous sulfate by the bacterium Thio-
References bacillus ferrooxidans. Biotechnology and Bioengineering,
149, 401–405.
Blais, J. F, Auclair, J. C, & Tyagi, R. D. (1992). Cooperation Lee, J.-U., Kim, S.-M., Kim, K.-W., & Kim, I. S. (2005).
between two Thiobacillus strains for heavy metal removal Microbial removal of uranium in uranium-bearing black
from municipal sludge. Canadian Journal of Microbiol- shale. Chemosphere, 59, 147–154.
ogy, 38, 181–187. Lizama, H. M., & Suzuki, I. (1991). Interaction of chalcopyrite
Boon, M., Snijder, M., Hansford, G. S., & Heijnen, J. J. (1998). and sphalerite with pyrite during leaching by Thiobacillus
The oxidation kinetics of zinc sulphide with Thiobacillus ferrooxidans and Thiobacillus thiooxidans. Canadian
ferrooxidans. Hydrometallurgy, 33, 137–152. Journal of Microbiology, 37, 304–311.

Environ Geochem Health

Schippers, A., Jozsa, P.-G., & Sand, W. (1996). Sulfur chem- extractants. ASCE Journal of Environmental Engineering,
istry in bacterial leaching of pyrite. Applied and Envi- 124, 639–645.
ronmental Microbiology, 62, 3424–3431. Stookey, L. L. (1970). Ferrozine—a new spectrophotometric
Silverman, M. P., & Ehrlich, H. L. (1964). Microbial formation reagent for iron. Analytical Chemistry, 42, 779–781.
and degradation of minerals. Advances in Applied Tuovinen , O. H. (2004). Biogeochemical transformations of
Microbiology, 6, 181–183. iron in bioleaching and acid mine drainage-impacted
Silverman, M. P., & Lundgren, D. G. (1959). Studies on the environments. Proceedings of the 8th Symposium
chemoautotrophic iron bacterium Ferrobacillus ferrooxi- on Waste Treatment and Recycling—Biotechnology,
dans. I. An improved medium and a harvesting procedure pp. 207–227.
for securing high cell yields. Journal of Bacteriology, 77, Tyagi , R. D., & Couillard D. (1989). Bacterial leaching
642–647. of metals from sludge. In P. E. Cheremisinoff (Ed.)
Sreekrishnan, T. R., Tyagi, R. D., Blais, J. F., & Campbell, P. Encyclopedia of Environmental Control Technology 3,
G. C. (1993). Kinetics of heavy metal bioleaching from (pp. 557–591). Houston: Gulf Publishing.
sewage sludge. I. Effects of process parameters. Water Wasay, S. A., Barrington, S. F, & Tokunaga, S. (1998).
Research, 27, 1641–1651. Remediation of soils polluted by heavy-metals using salts
Steele, M. C., & Pichtel, J. (1998). Ex-situ remediation of of organic-acids and chelating-agents. Environmental
a metal-contaminated superfund soil using selective Technology, 19, 369–379.