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Microbial iron reduction as a method for immobilization of a low

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DOI: 10.1016/j.jenvman.2018.04.023

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Contents lists available at ScienceDirect

Journal of Environmental Management

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journal homepage: www.elsevier.com

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Research article

Microbial iron reduction as a method for immobilization of a low concentration of

dissolved cadmium
Chaolei Yuana, Tongxu Liua, Fangbai Lia, ∗, Chengshuai Liua, Huanyun Yua, b, Weimin Suna, Weilin Huanga, c

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a
Guangdong Key Laboratory of Integrated Agro-environmental Pollution Control and Management, Guangdong Institute of Eco-environmental Science & Technology, Guangzhou
510650, China
b
Guangzhou Key Laboratory of Environmental Exposure and Health, School of Environment, Jinan University, Guangzhou 510632, China
c
Department of Environmental Sciences, Rutgers University, New Brunswick, 08901, NJ, USA

ARTICLE INFO ABSTRACT

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Article history: Much attention has been paid to the relationship between microbial iron reduction and the behavior of cad-
Received 12 December 2017 mium (Cd) recently, but most previous research has employed unrealistically high Cd concentrations (e.g.,
Received in revised form 24 February 2018 2–55 mg L−1) and has failed to consider the effects of iron oxides and microbial cells together. We investi-
Accepted 5 April 2018
gated the reduction of lepidocrocite by Shewanella oneidensis MR-1 in the presence of a low concentration
Available online xxx
of Cd using batch reactor systems. The results showed that with 422 μg L−1 added dissolved Cd2+, an initial
137 μg L−1 decrease in aqueous Cd occurred due to adsorption onto lepidocrocite and that the further removal
CT
Keywords: of remaining aqueous Cd occurred only in the system containing bacteria. This further decrease in aqueous Cd
Cadmium was unlikely to be caused by mineral transformation because the microbial reduction of lepidocrocite resulted
Immobilization in particle-size-increased (thus, specific-surface-area-decreased) lepidocrocite, and unlikely to be caused by
Iron oxide the pH increase to 7.4 induced by iron reduction either because a pH-adsorption edge suggested that at pH 7.4,
Microbial cell less than 60% of aqueous Cd can be adsorbed by lepidocrocite in the reactors. An adsorption isotherm showed
Microbial iron reduction
a significant Cd adsorption capacity by S. oneidensis MR-1 cells, and we therefore attributed the further Cd
removal to adsorption by S. oneidensis MR-1 cells. The results suggest that a realistically low concentration
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of Cd can be immobilized during microbial iron reduction by adsorption on iron oxides and microbial cells.
© 2018.

1. Introduction probably immobilized in magnetite that formed during microbial

Fe(III)-mineral reduction. Thus, iron reduction may not result in the
Cadmium (Cd) is a widespread metal pollutant in agricultural soils, release of Cd and the effects of iron reduction on Cd behavior need
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and exposure to Cd in ways such as rice consumption can increase further investigation.
the risk of osteoporosis, renal diseases, and cancer (Maret and Moulis, In addition to iron oxides, microbial cells also have a significant
2013; Muehe et al., 2013a). Microbial iron reduction is an impor- capacity to adsorb metals including Cd, owing to various functional
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tant process in saturated soils such as rice paddy soils (Yu et al.,
2016). Iron oxides can adsorb Cd due to their large surface area, and
therefore, the reductive dissolution of iron oxides may lead to the
release of Cd (Muehe et al., 2013a; Yu et al., 2016). On the other
hand, some studies have suggested that secondary mineralization dur-
ing iron reduction could immobilize Cd. Li et al. (2016) investigated
the reduction of Cd-loaded polyferric flocs by Shewanella oneiden-
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groups (e.g., carboxyl, sulfhydryl, phosphoryl, and amide groups) on
cell walls (Chang et al., 1997; Ha et al., 2010; Kaulbach et al., 2005;
Mishra et al., 2010; Yu and Fein, 2015). However, the effects of
microbial cells on Cd immobilization have hardly been considered
together with those of iron oxides during microbial iron reduction.
Moreover, the Cd concentrations in the experimental systems of pre-
vious studies were usually too high (e.g., 2–55 mg L−1 (Li et al., 2016;

sis MR-1 and found that Cd2+ was first released into the solution and
was subsequently bound by secondary iron minerals that had formed.
During the microbial reduction of ferrihydrite in the presence of Cd
in solution, Cd was adsorbed to and/or co-precipitated within sec-
ondary carbonate minerals (Muehe et al., 2013a). Further, after incu-
bation of a Cd-contaminated soil under reducing conditions with the
addition of organic matter, Muehe et al. (2013b) found that Cd was
∗
Corresponding author.
Email address: cefbli@soil.gd.cn (F. Li)
Muehe et al., 2013a)) compared to general Cd-contaminated soils.
For example, total Cd concentration in European soils ranges between
0.06 and 0.6 mg kg−1 (Soco Project Team, 2009), 95% of agricultural
soils in the USA have a Cd concentration < 0.78 mg kg−1 (Holmgren
et al., 1993), and in China, most of the Cd-contaminated soils con-
tains < 1 mg kg−1 Cd (Wang et al., 2015). Thus, more attention should
be paid to the fate of Cd that occurs in environmentally realistic con-
centrations (<1 mg kg−1 Cd).
During anaerobic respiration, iron-reducing bacteria use Fe(III)
in iron oxides as an electron acceptor and produce Fe2+. The bio-
genic Fe2+ can induce the transformation of iron oxides (Hansel et al.,

https://doi.org/10.1016/j.jenvman.2018.04.023
0301-4797/ © 2018.
2 Journal of Environmental Management xxx (2018) xxx-xxx
2003). The co-existence of metal cations such as Cd2+ can influence
microbial reduction and transformation of iron oxides. First, if the
co-existing metal cations are toxic to cells, the activity of iron-reduc-
ing bacteria and thus Fe2+ generation will be constrained (Trevors et
al., 1986). Second, recent studies have shown that concomitant metal
cations can inhibit Fe(II)-catalyzed mineral transformation, and the
possible mechanisms include the competition between Fe2+ and con-
comitant metal cations for adsorption sites on the surface of iron ox-
ides (Liu et al., 2016). Lepidocrocite (γ-FeOOH) is an iron oxide-hy-
droxide that is found in many saturated soils (Schwertmann, 1988).
Laboratory incubation experiments show that the microbial reduction
of lepidocrocite can produce various secondary minerals, such as mag-
netite, green rusts, vivianite, and goethite, depending on many fac-
tors such as the iron reduction rate, cell density, and medium com-
position (Bae and Lee, 2013; Li et al., 2012; O'Loughlin et al., 2010;
Ona-Nguema et al., 2002; Zegeye et al., 2007, 2010). However, how
the presence of Cd affects the transformation of lepidocrocite during
microbial reduction is not fully understood.
In this study, we investigated the microbial reduction of lepi-
docrocite in the presence of a low concentration (422 μg L−1) of Cd
using batch reactor systems. The spatial distribution of Cd was deter-
mined by chemical extraction, and the mineral phase was examined
using various physical tools. Adsorption isotherms and a pH-adsorp-
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tion edge were built to help distinguish the contributions by iron oxide
and microbial cells to Cd immobilization. Our hypotheses are that (1)
microbial lepidocrocite reduction can immobilize a low concentration
of aqueous Cd, (2) adsorption by microbial cells contributes signifi-
cantly to Cd immobilization, and (3) the presence of a low concentra-
tion of Cd does not affect lepidocrocite transformation during micro-
bial reduction.
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2. Materials and methods
2.1. Lepidocrocite and Shewanella oneidensis MR-1
Lepidocrocite was synthesized by mixing solutions of iron(II)
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chloride tetrahydrate, hexamethylene tetramine, and sodium nitrite ac-
cording to Hall et al. (1995). The product was confirmed by X-ray dif-
fraction (XRD) as lepidocrocite. Shewanella oneidensis MR-1 (here-
after MR-1), a well-studied iron-reducing bacterium (Li et al., 2016),
was purchased from American Type Culture Collection (ATCC®
700550™). MR-1 is facultative and can survive and proliferate in both
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aerobic and anaerobic conditions (Venkateswaran et al., 1999).
2.2. Incubation setup
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There were three batch reactor systems: γ-FeOOH incubated with
Cd2+, γ-FeOOH incubated with MR-1, and γ-FeOOH incubated with
MR-1 and Cd2+, which are referred to hereafter as systems Fe+Cd,
Fe+M, and Fe+M+Cd, respectively. The incubation was slightly mod-
ified after Liu et al. (2011). MR-1 cells were grown in lysogeny broth
containing 10 g L−1 tryptone, 5 g L−1 yeast extract, and 10 g L−1 NaCl at
30 °C for 16 h under aerobic conditions to allow the bacteria to rapidly
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proliferate. Then, the cells were harvested and washed three times
by centrifugation and re-suspension with a PIPES buffer medium
(adjusted to pH 7.0 using KOH; autoclaved) containing 9.07 g ho-
mopiperazine-1,4-bis(2-ethanesulfonic acid) (PIPES), 10 ml vitamin
solution, 10 ml mineral solution, and 4 ml 5 M sodium lactate so-
lution in 1 L (Lovley, 2013). A total of 0.0711 g lepidocrocite was
weighed into 50-ml serum bottles. The bottles were sealed and au-
toclaved for 20 min at 121 °C, after which lepidocrocite remained un-
changed as confirmed by XRD. Autoclaved PIPES buffer medium
was then added to the bottles, and the suspension was bubbled with N2
(passed by a 0.22-μm membrane) for 40 min to repel oxygen. A 2-ml
MR-1 cell suspension (OD600 = 2.0) was added for the systems with
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bacteria, and the systems with Cd2+ contained 422 μg L−1 Cd2+ (added
as CdCl2 solution). This quantity of Cd was adopted because accord-
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ing to the environmental quality standard for soils in China, to en-
sure agricultural production and human health, the upper limit for the
concentration of total soil Cd is 0.3 mg kg−1 for soils with pH ≤ 7.5 or
0.6 mg kg−1 for soils with pH > 7.5 (Wang et al., 2015). The total vol-
ume of the suspension was 40 ml per bottle for all the systems. The
oxygen-repelled bottles were then sealed with rubber stoppers and alu-
minum lids and incubated in a shaker at 150 rpm and 30 °C in the dark.
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2.3. Analyses for Cd distribution and mineral transformation
Three reactors of each system were transferred into a glove box
at predesigned time points. A 2-ml suspension was transferred to a
15-ml tube and mixed with 8 ml 0.5 M HCl end-over-end for 1 h. The
mixture was then filtered with a 0.45-μm membrane, and the filtrate
(HCl extract) was collected. The remaining 38-ml suspension was
also passed through 0.45-μm membrane filters, and both the filtrate
(aqueous fraction) and mineral were collected. The concentrations of
Fe2+ and Cd2+ in the filtrates were measured via 1,10-phenanthroline
colorimetric assay and inductively coupled plasma optical emission
spectrometry (ICP-OES) (Optima 8000, PerkinElmer, USA), respec-
tively. The difference between the concentrations of Fe2+ or Cd2+ in
the HCl extract and aqueous fraction was assigned as 0.4 M HCl-ex-
tractable Fe2+ or Cd2+. The collected mineral was air-dried in the glove
box. The mineral in triplicate reactors was combined and then ana-
lyzed by XRD, infrared spectroscopy (IR), X-ray photoelectron spec-
troscopy (XPS), transmission electron microscopy (TEM), and Möss-
bauer spectroscopy. Details of the physical analyses are provided in
the supplementary materials.
2.4. pH-adsorption edge and adsorption isotherms
The pH-adsorption edge for Cd adsorption on lepidocrocite was
determined as follows. A 20-ml autoclaved PIPES medium with
422 μg L−1 Cd2+ and various pH between 6.1 and 7.5 (the useful pH
range of PIPES) was added to 50-ml polyethylene tubes each contain-
ing 0.0356 g lepidocrocite. After ultrasonic dispersion, the tubes were
shaken at 180 rmp and 30 °C in the dark for 24 h and were then cen-
trifuged at 7200 g for 5 min. The supernatants were collected by pass-
ing through a 0.22-μm membrane for measurements of pH and Cd
concentrations. An isotherm for Cd adsorption on lepidocrocite was
obtained by incubating 0.0356, 0.0533, 0.0800, 0.1067, and 0.1778 g
lepidocrocite in 20 ml PIPES medium with 422 μg L−1 Cd2+ and pH 7.0
in a similar way. Cd adsorption by Shewanella oneidensis MR-1 cells
was estimated by measuring the concentrations of aqueous Cd af-
ter incubating 2, 4, 6, and 8 ml MR-1 cell suspension (OD600 = 2.0) in
40 ml N2-bubbled PIPES buffer medium with 422 μg L−1 Cd2+ and pH
7.0 in 100-ml serum bottles for 48 h in a shaker at 150 rpm and 30 °C
in the dark. After 48 h, the cell density in the system did not increase
as revealed by OD600 because no electron acceptor was supplied for
the bacteria (i.e., no iron oxide was added to the serum bottles).
 Journal of Environmental Management xxx (2018) xxx-xxx 3

3. Results and discussion

3.1. Lepidocrocite reduction and transformation

In the system Fe+Cd that did not contain bacteria, the concen-
trations of aqueous and 0.4 M HCl-extractable Fe2+ remained con-

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sistently low. In contrast, in the system Fe+M, the concentrations
of aqueous and 0.4 M HCl-extractable Fe2+ increased to 167 and

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67 mg L−1 within 7 days, and in the system Fe+M+Cd, the concentra-
tions increased to 148 and 71 mg L−1 (Fig. 1). The increase in Fe2+
demonstrated the microbial reduction of lepidocrocite. After 7 days,
the Fe2+ concentrations became relatively stable. After 32 days, the pH
of the suspension in the system Fe+Cd was 7.01, and in the systems
Fe+M and Fe+M+Cd, it was 7.40 and 7.44, respectively, because iron
reduction is a proton-consuming process: CH2O + 4FeOOH + 8H+ →
CO2 + 4Fe2+ + 7H2O (Frommichen et al., 2004). The rate of microbial

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iron reduction in the first 7 days was slightly lower in the system with
added Cd2+ (1.25 mg Fe2+ day−1) than the system without added Cd2+
(1.34 mg Fe2+ day−1) (Fig. 1), probably due to the toxicity of Cd to mi-
crobial cells.
After 32 days, unlike in the system Fe+Cd, in the systems Fe+M
and Fe+M+Cd where microbial iron reduction occurred, the color of
the suspension became darker, and the iron oxides were attracted to a

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magnet (Fig. 2), suggesting the formation of magnetic material, prob-
ably magnetite, which has a black color (Bigham et al., 2002) and has
been reported as a product of microbial reduction of lepidocrocite in

Fig. 2. Appearance of reactors after 32 days which shows (a) the color of suspensions
and (b) attraction of mineral by a magnet. Fe+Cd, Fe+M, and Fe+M+Cd indicate sys-
tems with (i) γ-FeOOH and Cd2+, (ii) γ-FeOOH and MR-1, and (iii) γ-FeOOH, MR-1,
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and Cd2+, respectively. (For interpretation of the references to color in this figure leg-
end, the reader is referred to the Web version of this article.)

many studies (Li et al., 2012; Yuan et al., 2017; Zegeye et al., 2007).
However, XRD and IR analyses detected only lepidocrocite after 32
days in all the systems (Fig. 3). We then resorted to XPS and Möss-
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bauer spectroscopy, but we still could not detect the presence of mag-
netite (see supplementary figures). The reason is probably that pu-
tative magnetite accounted for a very small fraction of the iron ox-
ides and/or the magnetite was amorphous (Schwertmann and Cornell,
2000).
Various secondary minerals can be formed after the microbial re-
duction of lepidocrocite depending on reaction conditions as men-
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tioned before. Particularly, the transformation of lepidocrocite in the

presence of Fe2+ can also result in recrystallized lepidocrocite
(Pedersen et al., 2005, 2006). Using 55Fe labeling, Pedersen et al.
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(2005) observed rapid isotopic exchange between aqueous Fe2+ and

Fe in a 5-nm-sized lepidocrocite; after recrystallization, the mineral
phase remained as lepidocrocite, but the crystal size increased to 15
or 20 nm. In this study, TEM images also showed that the particle
size of lepidocrocite increased after microbial iron reduction (Fig. 4).
This is corroborated by the narrowed XRD peaks of lepidocrocites in
the systems Fe+M and Fe+M+Cd where iron reduction occurred com-
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Fig. 1. Time-dependent concentrations of aqueous and 0.4 M HCl-extractable Fe(II) in
batch reactor systems with (a) γ-FeOOH and Shewanella oneidensis MR-1 (hereafter
MR-1) and (b) γ-FeOOH, MR-1, and Cd2+. Error bars represent standard deviations.
pared to the system Fe+Cd where iron reduction did not occur (Fig.
3a), considering that narrowed XRD peaks indicate increased crys-
tal sizes (Schwertmann and Cornell, 2000). Taken together, our data
suggest that in this study, microbial iron reduction probably induced
the recrystallization of lepidocrocite, with crystal-size-increased lep-
idocrocite as the major product and magnetite a very minor product.
The presence of Cd did not have a significant effect on the iron oxide
transformation during microbial reduction in this study (Fig. 3), which
confirmed our third hypothesis, probably due to the very low Cd con-
centration (Cd/Fe = 0.019 mol.%).
4 Journal of Environmental Management xxx (2018) xxx-xxx
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Fig. 3. XRD and IR patterns of minerals on day 32. Lepidocrocite was the only identi-
fiable mineral in all the systems.
3.2. Cd immobilization and mechanisms
In the system Fe+Cd, at the first sampling time point that was
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approximately 2 h after mixing lepidocrocite with 422 μg L−1 dis-
solved Cd2+, the concentration of aqueous Cd2+ was 285 μg L−1 (i.e.,
decreased by 137 μg L−1) while the concentration of 0.4 M HCl-ex-
tractable Cd2+ was 140 μg L−1 (Fig. 5a). This indicated rapid adsorp-
tion of 140 μg L−1 dissolved Cd onto lepidocrocite in 2 h because 0.4 M
HCl mainly extracts Cd adsorbed on the solid phase in the system
(Jeon et al., 2003), and the great Cd adsorption capacity of lepi-
docrocite (113.6 μg Cd g−1 lepidocrocite maximum) was demonstrated
by an adsorption isotherm (Fig. 6a). After 2 h, the concentrations of
aqueous and 0.4 M HCl-extractable Cd2+ became relatively stable in
the system Fe+Cd.
In the system Fe+M+Cd, the concentration of aqueous Cd2+ at the
first sampling time point was even lower (207 μg L−1) compared to the
system Fe+Cd (Fig. 5), probably due to additional Cd2+ adsorption by
MR-1 cells. The aqueous Cd2+ then decreased dramatically from 207
to 12 μg L−1 within 2 days and further to under the detection limit on
day 7. Correspondingly, the concentration of 0.4 M HCl-extractable
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Cd2+ increased to 385 μg L−1 after 7 days. Namely, after an initial de-
crease by approximately 200 μg L−1, the remaining 207 μg L−1 aqueous
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Cd transferred into adsorbed Cd during microbial iron reduction. This
adsorption was unlikely to be caused by mineral transformation be-
cause after 32 days, the particle size of lepidocrocite increased, which
leads to decreased specific surface area. The increase in the pH of
the suspension to 7.4 after 32 days could not explain the further de-
crease in aqueous Cd either, because a pH-adsorption edge suggested
that at pH 7.4, less than 60% of aqueous Cd could be adsorbed by
lepidocrocite in the reactors (Fig. 7). Thus, we speculated that the
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remaining 207 μg L−1 aqueous Cd was adsorbed by MR-1 cells. This
was supported by the Cd adsorption capacity of MR-1 cells (9.3 μg Cd
ml−1 MR-1 cells (OD600 = 2.0) maximum) as shown by an adsorption
isotherm (Fig. 6b). And Cd adsorption by MR-1 cells has also been re-
ported by other researchers (Ha et al., 2010; Mishra et al., 2010; Yu
and Fein, 2015). Considering microbial regeneration during iron re-
duction, for instance, Muehe et al. (2013a) observed that the cell num-
ber of iron-reducing bacteria increased by approximately two orders
of magnitude after 7 days during ferrihydrite reduction, the adsorp-
tion of Cd by MR-1 could be significant. The probable increase in the
MR-1 cell number after 7 days in this study could be suggested by the
generation of Fe2+ in reactor systems. The results, therefore, supported
our first hypothesis that microbial lepidocrocite reduction can immo-
bilize a low concentration of aqueous Cd and our second hypothesis
that adsorption by microbial cells contributes significantly to Cd im-
mobilization. The change in the spatial distribution of Cd during the
microbial reduction of lepidocrocite in this study is shown in a con-
ceptual model (Fig. 8).
During microbial iron reduction in this study, there was no ini-
tial increase and subsequent decrease in aqueous Cd, as shown for
the cases reported by Li et al. (2016) and Muehe et al. (2013a). In
the experiment conducted by Li et al. (2016), the substrate for She-
wanella oneidensis MR-1 was Cd-loaded polyferric flocs (i.e., Cd was
not added in dissolved form), and thus, Cd was released to the solution
after reductive dissolution of the solid phase. In the study by Muehe
et al. (2013a), within 9 days, approximately 40% of Fe in ferrihydrite
was transformed to Fe(II), so a significant amount of Cd that was ad-
sorbed on ferrihydrite shortly after the addition of aqueous Cd could
be released into the solution again, which was amplified by the large
concentrations of added Cd (11 or 55 mg L−1). The greater extent of
iron reduction observed by Muehe et al. (2013a) compared to the one
in this study (<15% after 32 days) might be attributed to the different
types and concentrations of iron oxides and iron-reducing bacteria em-
ployed.
3.3. Environmental implications
This study demonstrated that during microbial iron reduction, ad-
sorption by 20 mM lepidocrocite can immobilize approximately
one-third of 400 μg L−1 dissolved Cd2+, and the remaining Cd2+ could
be adsorbed by bacterial cells (Fig. 8). This could provide insight for
the management of realistic Cd-polluted paddy soils wherein micro-
bial iron reduction is an important biogeochemical process. First, an
extended flooding period is recommended for rice cropping in Cd-pol-
luted areas, and the effectiveness has been attributed to increased soil
pH and the formation of CdS (Fulda et al., 2013; Li et al., 2017; Sun
 Journal of Environmental Management xxx (2018) xxx-xxx 5

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Fig. 4. TEM images of lepidocrocites after 32 days in batch reactor systems with (a) γ-FeOOH and Cd2+, (b) γ-FeOOH and MR-1, and (c) γ-FeOOH, MR-1, and Cd2+.

et al., 2007). This study highlighted the potential contributions from

iron oxides and especially their microbial reducers, considering that
iron oxides are abundant (Loeppert and Inskeep, 1996) and iron-re-
ducing bacteria are widespread in paddy soils (Yu et al., 2016). Sec-
ond, the stimulation of microbial iron reduction by, for example,
adding iron oxides or electron donors (e.g., organic matter) to the soil

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may be useful to reduce Cd bioavailability in paddy soils that contain
a relatively low concentration of Cd. To answer this question, further
soil and field experiments are required.

Acknowledgements
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This work was supported by the National Natural Science Founda-
tion of China (41420104007, 41601239), the China Postdoctoral Sci-
ence Foundation (2016M600644), the “Pearl River Talents” Postdoc-
toral Program of Guangdong Province, the High-level Leading Talent
Introduction Program of GDAS, and the National Key Research and
Development Program of China (2016YFD0800703). Huanyun Yu
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also acknowledges funding from the Guangzhou Key Laboratory of

Environmental Exposure and Health (GZKLEEH201604). We thank
Dr. Chengyu Chen for help in analyzing data of adsorption isotherms.

Appendix A. Supplementary data

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Supplementary data related to this article can be found at https://

doi.org/10.1016/j.jenvman.2018.04.023.
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Fig. 5. Time-dependent concentrations of aqueous and 0.4 M HCl-extractable Cd in

batch reactor systems with (a) γ-FeOOH and Cd2+ and (b) γ-FeOOH, MR-1, and
Cd2+. Error bars represent standard deviations.
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6 Journal of Environmental Management xxx (2018) xxx-xxx

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Fig. 6. Isotherms of Cd adsorption at pH 7.0 by (a) lepidocrocite and (b) Shewanella oneidensis MR-1 cells (OD600 = 2.0). The Langmuir isotherm, q = qmaxCe/(Kd + Ce), was used
to describe the adsorption equilibrium, where q is adsorption density, qmax the maximal adsorption capacity (113.6 μg Cd g−1 lepidocrocite for (a) or 9.3 μg Cd ml−1 MR-1 cells
(OD600 = 2.0) for (b)), Ce the equilibrium concentration of adsorbate (Cd), and Kd dissociation constant (25.0 μg L−1 for (a) or 27.5 μg L−1 for (b)).

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Fig. 7. Adsorption of Cd by 20 mM lepidocrocite in the PIPES (homopiper-

azine-1,4-bis(2-ethanesulfonic acid)) medium as a function of pH between 6.1 and 7.5
(the useful pH range of PIPES). Error bars represent standard deviations.
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Fig. 8. A conceptual model showing change in spatial distribution of Cd during microbial reduction of lepidocrocite in this study. After around 2 h, about half of the 422 μg L−1
added dissolved Cd2+ was adsorbed by lepidocrocite and bacterial cells. After 32 days, microbial reduction caused Fe2+ generation and lepidocrocite transformation to particle-size-in-
creased lepidocrocite. The remaining dissolved Cd2+ was immobilized via adsorption on bacterial cells which probably proliferated.
 Journal of Environmental Management xxx (2018) xxx-xxx 7

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