Journal of Environmental Management 212 (2018) 115e120

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Journal of Environmental Management

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Short communication

Adsorption of phenanthrene by earthworms - A pathway for

understanding the fate of hydrophobic organic contaminants in
soil-earthworm systems
Zhiming Shi, Fangfang Zhang, Congying Wang*
College of Environmental and Resource Sciences, Shanxi University, Taiyuan, 030006, PR China

a r t i c l e i n f o a b s t r a c t

Article history: The fate of hydrophobic organic contaminants (HOCs) in soil-earthworm systems is an important
Received 30 October 2017 foundation for soil pollution risk assessment and pollution control. Equilibrium partitioning is consid-
Received in revised form ered to be the main mechanism by which HOCs enter earthworms and, as such, a mechanistic model has
26 January 2018
been developed to estimate the earthworm-water partition coefficients (Kw-w). In the present study, the
Accepted 28 January 2018
adsorption of phenanthrene (PHE) by earthworm tissue was investigated to evaluate the validity of the
mechanistic models. Results revealed that Kw-w derived from the mechanistic model (346.90) was lower
than that derived from the sorption experiments (410.76), indicating that apart from lipid fractions, other
Keywords:
Earthworm
components in earthworms, such as protein fractions, might also play an important role in the
Mechanistic model adsorption of HOCs by earthworm. Besides, the difference between the mechanistic model for earth-
Adsorption behaviour worm and partition-limited model used for plants are few, indicating that uptake and accumulation
Distribution of phenanthrene in earthworm mechanisms of HOCs by earthworms and plants are highly consistent internally and are, essentially
Vermiremediation actually identical. It is also suggested that environmental fate of HOCs in soil-soil biota systems is
dominated by their high hydrophobicity. Based on these conclusions, an improved mechanistic model for
predicting the uptake of organic contaminants by earthworms has been proposed, which needs to be
further evaluated. Furthermore, the feasibility of using vermiaccumulation in vermiremediation of soil
contaminated by HOCs was discussed.
The adsorption of PHE by earthworm sub-organism fractions (pre-clitellum, clitellum and post-
clitellum) and tissue fractions (body wall and gut) were also investigated to interpret the distribution
pattern of HOCs in earthworms. At the sub-organism level, the adsorption capacity of PHE by different
regions of the earthworm followed the order: post-clitellum > clitellum > pre-clitellum, meaning the
distribution of PHE along the earthworm contributes not only to their chemical composition but also to
the life activity of earthworms such as circular system. At the tissue level, the gut showed greater affinity
with PHE than that of the body wall indicating that the distribution of PHE is mainly due to chemical
components at the tissue levels. These results might provide additional understanding of the fate of
HOCs in soil-earthworm systems.
© 2018 Elsevier Ltd. All rights reserved.

1. Introduction badgers, thrushes and other predators (Jager, 1998). Vermir-

Earthworms are one of the most important soil fauna and play a
key role in terrestrial ecosystems (Peijnenburg and Vijver, 2009).
The accumulation of hydrophobic organic contaminants (HOCs) in
earthworms leads to the transfer of contaminants into the terres-
trial food chain/web, followed by the possible preying by moles,
* Corresponding author.
E-mail addresses: shizhiming@sxu.edu.cn (Z. Shi), 2824006559@qq.com
(F. Zhang), wangcongying@sxu.edu.cn (C. Wang).
emediation of organic polluted soils, which refers to the use of
earthworms for the removal of contaminants from soil or when
earthworms help to degrade non-recyclables, has recently drawn
attention as a possible innovative technology (Dendooven et al.,
2011; Gupta and Garg, 2009; Rodriguez-Campos et al., 2014).
Further, earthworms are important model organisms used to
indicate the status of soil health by investigating their responses to
various external stressors such as soil contaminants (Lankadurai
et al., 2015; Shi et al., 2013). Therefore, understanding the behav-
iour of uptake, accumulation, translocation and distribution of
HOCs by earthworm is fundamental for soil pollution ecological risk

https://doi.org/10.1016/j.jenvman.2018.01.079
0301-4797/© 2018 Elsevier Ltd. All rights reserved.
116 Z. Shi et al. / Journal of Environmental Management 212 (2018) 115e120

assessment and provide guidance for the practice application of experiment, earthworms were pre-incubated in mixtures of
vermiremediation. potting compost soil and cow dung in the dark at 22 ± 1
C for 2
HOCs may enter earthworms via epidermal uptake and dietary weeks.
uptake and then disperse within the earthworms. Epidermal up- Phenanthrene (PHE, purity 97%) was purchased from the Fluka
take is usually predominant over dietary uptake for hydrophobic Company, Germany. All reagents used in the study were of
chemicals that have a logKow < 5 (Kow is the octanol-water partition analytical grade.
coefficient) (Belfroid et al., 1995; Jager et al., 2003). The uptake of
HOCs by earthworms is thought to depend on the properties of the
contaminants, earthworm species, and soil types (Lanno et al.,
2.2. Preparation of dry earthworm, sub-organism fractions and
2004). The relationships between earthworm uptake and the
tissue fractions
physical-chemical properties of contaminants have been estab-
lished and have been described as linear between the logarithms of
Adult earthworms (each 200e300 mg, wet weight) with well-
earthworm bioconcentration factors (BCFs) and the logarithm of
developed clitellum were chosen randomly and placed on a moist
the Kow of contaminants. However, these relationships were
filter for 24 h to depurate their gastrointestinal tract. Gut-voided
formulated empirically (Jager, 1998; Raber et al., 1998). Afterwards,
earthworms were then rinsed with deionized water and softly
a mechanistic model was developed for estimating the bio-
dried with absorbent paper. Dry whole earthworm, sub-organism
concentration of organic chemicals in earthworms, based on the
fractions (pre-clitellum, clitellum and post-clitellum) and tissue
contaminant passive uptake by earthworms in a series of parti-
fractions (gut and body wall) powder were obtained according to
tioning processes among soil solids, soil water, and the resident
the methods established in our previous study (Shi et al., 2014).
organism's tissue (Hodson et al., 2011; Jager, 1998; Khan et al., 2011;
The dead earthworm or fractions were weighted and freeze-
Reiss et al., 2009).
dried using a vacuum freeze dryer (Alpha 1e2 LD plus, Sigma,
The mechanistic model assumes that an earthworm is an
German) for 72 h to determine the water content. Dried earthworm
inanimate container seeking to reach thermodynamic equilibrium
or fractions were then ground into powder using an agate mortar.
with its medium. Therefore, studying the adsorption of organic
contaminants by earthworm tissues is the key to understanding the
uptake process. However, until now, it seems that there has been a
lack of the quantitative evaluation of the partitioning coefficient 2.3. Adsorption experiments
between earthworms and water in equilibrium (Kw-w). Conse-
quently, the assumption of the mechanistic model that the earth- All sorption isotherms were obtained using a batch equilibration
worm's lipid fractions are the main storage pool of HOCs and that technique at 30 ± 1
C. The adsorption of PHE by earthworms or
other components, such as protein and carbohydrates, can be their parts was determined using the following approach: 20.0 mg
neglected, need further validation. If it is true, then Kw-w derived of dry earthworm powder, sub-organism fraction (pre-clitellum,
from the sorption experiment should be consistent with that of the clitellum, and post-clitellum regions) powder and tissue fraction
mechanistic model. (body wall and gut regions) powder were transferred into 40 mL
The distribution of HOCs in earthworm is usually considered the brown glass centrifuge tubes and 25 mL of 0.01 mol/L KNO3 solu-
result of partitioning between different chemical fractions, which is tion containing a certain concentration of PHE (nominal initial
not well defined between organs or regions or parts along the aqueous concentrations of 0.1, 0.25, 0.5, 0.7 and 1 mg/L according to
earthworm. Since different regions of an organism possess different the aqueous solubility of HPE at 25
C). Less than 0.1% methanol and
physiological functions, understanding the exact distribution of 0.02% NaN3 were also added to the tubes to avoid any cosolvent
HOCs in earthworm is very important to better clarify the toxic effects and prevent any possible biodegradation, respectively.
response mechanism (Escher et al., 2011; Escher and Hermens, Tubes were shaken at 150 rpm for 36 h according to the preliminary
2004). In previous studies, application of the earthworm accumu- kinetic study. Samples were prepared in triplicate for each treat-
lation experiment and earthworm fractionation strategy, ment. Control samples containing PHE but no sorbent were pre-
confirmed that phenanthrene (PHE) in earthworm tissue level and pared to determine solute loss possibly occurring due to handling,
sub-organism levels is heterogeneously distributed (Shi et al., volatilisation and degradation.
2014). However, it is difficult to explain this heterogeneous distri-
bution. If entry of PHE into earthworm is mainly through the body
wall as a function of the partitioning processes, the distribution of
2.4. Determination of PHE in water
PHE in the earthworm should be homogenous across the different
fractionation levels and be consistent with that of the adsorption
Tubes were centrifuged at 4000 rpm for 10 min and 0.5 mL of
pattern in the different sub-organism regions.
supernatants was diluted with 0.5 mL of HPLC grade methanol. The
Based on these statements, the study the adsorption of HOCs by
supernatant was then filtered using a 0.22 mm Teflon filter mem-
earthworm tissues is important for understanding the fate of HOCs
brane. A Shimadzu HPLC (LC-20A, Japan) equipped with an
in soil-earthworm systems. Specifically, the adsorption of PHE, a
UVevisible detector and a reverse-phase C18 chromatographic
representative HOCs, to the whole earthworm, sub-organism
column (25 cm  4.6 mm, 5 mm particle diameter) was used for PHE
fractions and tissue fractions was investigated to validate the
quantification (Ma et al., 2012; Zhang and Zhu, 2009).
assumption of the mechanistic model and to explain the internal
distribution behaviour of PHE within earthworms.

2. Material and methods 2.5. Statistical analyses

2.1. Test earthworm and chemicals The amount of sorped PHE was calculated based on the differ-
ence between the solution concentration and the total amounts of
The Eisenia fetida (Savigny 1826) earthworm used in this study PHE added to the solution. Slopes of sorption isotherms (Kw-w)
were purchased from a commercial supplier. Prior to the were calculated from the linear part of each isotherm.
 Z. Shi et al. / Journal of Environmental Management 212 (2018) 115e120
3. Results and discussion
3.1. Water content in the whole earthworms and their different
fractions
The water content of earthworms, listed in Table 1, is important
data for the following calculation and discussion. The results indi-
cate that the water content of the whole earthworm body obtained
through freeze-drying was 82.22%, which is consistent with other
reports (Jager, 1998). Further, the results prove that freeze-drying
method is valid for the determination of the water content of
earthworm tissues. As for the sub-organism level, the water con-
tent was similar to that of the whole body, because the sub-
organism fractions were obtained by simply slicing and, thereby
the water loss in the process was negligible. Meanwhile at the
tissue level, the water content in the body wall and gut was lower
than that in the whole body and sub-organism fractions. Our pre-
vious study showed that the mass distribution proportion of body
wall, gut and body fluid is approximately 5:3:2 and, therefore, the
body fluid accounts for about 25% of the total water in earthworms
(Shi et al., 2014). Since the tissue fractions were obtained by
excluding the body fluid, the observed decrease in water content in
the organic fraction was expected.
3.2. Adsorption of PHE by the whole earthworm
Compared with bioaccumulation experiment, adsorption
experiment using earthworm dry powders will provide a rapid
equilibrium time since the large surface areas which is within 36 h
according to the preliminary kinetic study while the former needed
about 14e28 d (Ma et al., 2012; Shi et al., 2013). As a life system,
despite there are mechanisms of adaptation, active removal or
decontamination in worms bodies, when came to discuss the up-
take and bioaccumulation of HOCs, special some POPs in organisms,
their metabolism and decontamination are little compared with
the amount of uptake and bioaccumulation and are generally
neglected (Gomez-Eyles et al., 2011; Liao et al., 2015; Zhang and
Zhu, 2009). In earthworms, as far as we know, there is no clear
evidence showing that contribution of metabolism of PAHs in
earthworm is significant when discussing the burden/
Table 1
Water content in the earthworms and their different fractions.
Fractionations Fractions Water content (%)
Earthworm body e 82.22（0.65）
Sub-organism level Pre-clitellum 83.03（0.33）
Clitellum 80.74（0.42）
Post-clitellum 83.07（0.13）
Tissue level Body wall 77.47（0.25）
gut 60.74（0.39）
Note: Data in parentheses are standard deviations.
Table 2
Isotherms of PHE to whole body and different regions of earthworm and corresponding adsorption coefficients.
Fractions Adsorption isotherms
Whole body Qe ¼ 2282.6 Ce
Pre-clitellum Qe ¼ 1061.6Ce
Clitellum Qe ¼ 1815.6Ce
Post-clitellum Qe ¼ 2816.1Ce
Body wall Qe ¼ 3545.4Ce
Gut Qe ¼ 2180.8Ce
Note: *water content was adopted from Table 1.
Qe (mg/kg) is the sorption amount of phenanthrene on sub-organism fractions at equilibrium concentration (Ce, mg L1), Kr is the partition coefficient between different
fractions of earthworm and water.
117
bioaccumulation of PAHs. Besides, the process of HOCs in soil entry
earthworm is a series of equilibrium-partitioning processes be-
tween soil solid, soil water and earthworm phases and the parti-
tioning of HOCs between soil water and earthworm tissue are one
of the key processes (Jager et al., 2003). Therefore, the adsorption
experiment using earthworm dry powders will be a valuable tool
for exploring the behaviors of HOCs in earthworm-soils systems.
The determination of the adsorption of PHE by the earthworm
body was conducted by batch sorption experiments, with the
sorption isotherms shown in Table 2. The sorption isotherms were
highly linear over the range of concentrations, with R2 values
greater than 0.99, indicating the predominant role of partitioning in
sorption (Ling et al., 2009). Despite few studies on the sorption of
exotic chemicals to earthworms, many previous studies have
observed a linear partitioning of exotic chemicals to organic matter.
For example, the environmental fate of HOCs in soil-plant systems
has been widely studied and the linear partitioning of PAHs to
ryegrass roots has been proven (Zhang and Zhu, 2009). Due to the
linear characteristic of the sorption isotherms in the present study,
the partition coefficient of PHE for earthworms could be obtained
from the slopes. The partition coefficient (Kw-w) of PHE between
earthworms and water was 410.76 mg kg 1 after accounting for the
water content of the earthworms (82.22 ± 0.65%).
Based on the mechanistic theory, earthworm BCFs can be pre-
dicted using the following equation (Jager, 1998):
Cworm Kww Fwater þ Flip Klip
BCFs ¼ ¼ ¼ ðL=kg wormÞ (1)
Cwater rworm rworm
where Cworm and Cwater are the concentrations of HOCs in earth-
worms and soil solution, respectively. Flip and Fch are the volume
fractions of lipids and water in earthworms, respectively. rworm is
the bulk density of the earthworm. Klip is the lipids-derived parti-
tion coefficient.
Therefore, a predictive equation for organic contaminant con-
centrations in earthworm can be described according to the
following equation (assumption that rworm ¼ 1) (Jager, 1998):
 
Cworm ¼ Cwater Fwater þ Flip Klip (2)
Kw-w can be calculated using the following equation:
Kww ¼ Fwater þ Flipid Klip  ðL=LÞ (3)
This model accounts for the role of water and lipid fractions but
neglects other components such as protein and carbohydrate
fractions during earthworms in the partitioning processes of HOCs.
Octanol was used to mimic biological lipids in the contaminant
partitioning because of the unavailability of Klip, and, as such, the
Klip of contaminants were assumed to be the same as the corre-
sponding octanolewater partition coefficients (Kow). In the present
study, volume fractions were required. According to the literature
R2 Kr (L kg1, dry weight) Kr (L kg1, fresh weight*)
0.990 2282.6 410.76
0.98 1061.6 180.47
0.99 1815.6 363.12
0.99 2816.1 478.74
0.97 3545.4 815.4
0.98 2180.8 872.3
118 Z. Shi et al. / Journal of Environmental Management 212 (2018) 115e120
and our study (Shi et al., 2014), the lipid volume fraction was 0.012,
the water volume fraction was 0.82 and Kow of PHE is 28,840.31.
Therefore, the partition coefficient derived from the mechanistic
model was 346.90 (Km), which is lower than that from the
adsorption experiments (Kw-w ¼ 410.76).
This difference between the above mentioned two values sug-
gests that lipid fractions might not be sufficient to explain the
sorption of HOCs to earthworms and that the adsorption capacity of
PHE might be underestimated. Protein accounts for a predominant
part of the dry weight fraction. It was reported that Eisenia foetida
had a high protein content in the range of 54.6e71.0% of dry matter,
which was 10 times that of lipids (Medina et al., 2003; Zhenjun
et al., 1997). As a result, although protein might have a relatively
low affinity capacity, it may also play an important role in the
adsorption and uptake of organic contaminants by earthworms. In
fact, the Kw-w derived from the mechanistic model is just slightly
lower than that from sorption experiments.
Besides, it is worth noting that Chiou et al. (2001) proposed a
partition-limited model to predict the uptake of organic contami-
nants by plants, where the theory was based on the passive
contaminant uptake by plants as a series of local partition processes
between various plant organic components and water (or soil
water) and is expressed by the following equation (Chiou et al.,
2001):
 
Cpt ¼ Cw flip Klip þ fch Kch þ fpw
where Cpt and Cw are the concentrations of organic contaminant in
plant and soil solutions, respectively. Klip and Kch are lipid-derived
and carbohydrate-derived partition coefficients, respectively. flip,
fpw and fch are the weight fraction of lipids, water and carbohy-
drates, respectively.
By comparing equation (1) with equation (4), a remarkable
similarity was found when the contribution of carbohydrates in the
uptake of HOCs by plants was ignored. It indicates that although
huge biological differences exist between lower animals and higher
plants, uptake and accumulation mechanisms of HOCs by these
organisms are consistent internally and are actually essentially
identical. It also suggests that the environmental behaviors of HOCs
in soil-soil biota systems is dominated by their high
hydrophobicity.
Furthermore, the partition-limited model, which predicts the
uptake of organic contaminants by plants, was improved by
maintaining the integral roles of carbohydrate and lipid fractions
on the adsorption of organic contaminants to plants (Zhang and
Zhu, 2009). Similarly, in the present study, the underestimation of
Kw-w by the mechanistic model was confirmed. Therefore, the
mechanistic model for predicting earthworm accumulation of
HOCs should also be optimised by integrating the role of other
chemical components of earthworms, as presented in the following
equation:
 
Cworm ¼ Cwater Fwater þ Flip Klip þ Fpro Kpro
where Fpro is the volume fraction of protein and Kpro is the protein-
derived partition coefficient.
3.3. Adsorption of PHE by the earthworm sub-organism fractions
The mechanistic model did not concern the distribution of HOCs
in earthworms, but it hinted at the notion that the distribution
mechanism is governed by the equilibrium partitioning of HOCs
between the water fraction and other organic phases within
earthworms, i.e. the chemical components are the main
determining factors in the equilibrium partitioning process. Then
the question arises as to whether other earthworm life activities
apart from excretion, growth dilution and metabolism ect., are
contributing to the distribution of HOCs in earthworms. To address
the issue, batch sorption experiments for earthworm sub-organism
fractions (pre-clitellum, clitellum and post-clitellum) were carried
out and the isotherms parameters are shown in Table 2. As is
shown, the adsorption of PHE on earthworm sub-organism frac-
tions can be linearly correlated, with R2 values greater than 0.98,
and the sub-organism fraction-water partition coefficients could be
obtained from the slopes of the sorption isotherms. Partition co-
efficient (based on dry weight) in Table 2 suggest that the
adsorption capacity of PHE by different regions of earthworms
follow the order of post-clitellum > clitellum > pre-clitellum. Such
results were not consistent with the actual distribution pattern of
PHE in earthworm sub-organism levels. Our previous study found
that the distribution of PHE along earthworms is homogeneous
when they have a low body burden (Shi et al., 2014). This means
that earthworm life activities actually participate in the distribution
of HOCs along earthworms. Earthworms have a very simple circu-
latory system with two blood vessels (ventral and dorsal blood
vessels) running the length of the body (Gerard, 1985). Therefore, it
can be inferred that the circulatory system are might be also
responsible for the distribution of HOCs in earthworm sub-
organism levels.
(4)
3.4. Adsorption of PHE by earthworm tissue fractions
Earthworm tissues were divided into body wall fractions and
gut fractions. There are obvious anatomical and chemical compo-
sition distinctions between these. Dry matter biomass of the body
wall is higher than that of the gut and body fluid and, hence, the
distribution of contaminants in these fractions should be different.
In the previous study, we found that the concentration of PHE in the
body wall is lower than that in the gut (Shi et al., 2014). However,
the study was carried out using living earthworms and we cannot
know for sure whether the life activities affected the distribution. In
the present study, a batch sorption technique for body wall and gut
was conducted to evaluate its sorption capacity, which ruled out
the effect of life activity on the distribution of PHE. The isotherm
parameters are shown in Table 2 and the slopes of the isotherms
denote the body wall-water and gut-wall partition coefficients. The
isotherms were also linear, with R2 values greater than 0.97. The gut
(based on fresh weight) showed greater affinity for PHE (gut-water
partition coefficients ¼ 872.3 L kg1) than that of the body wall
(body wall-water partition coefficients ¼ 815.4 L kg1). Regardless
of the earthworm life cycle, such differences may be due to dif-
ferences in the chemical composition between the gut and wall.
Moreover, the difference is consistent with the real distribution
pattern of PHE in the gut and body wall, which means that the
distribution of PHE is mainly attributed to the chemical composi-
tion at the tissue level (Shi et al., 2014). Since oral uptake can be
neglected when the logKow is low, the translocation processes of
(5)
HOCs at the tissue level is likely a result of the passive diffusion of
contaminants in soil solution into the earthworm body wall which
is then further partitioned into the gut.
4. Environmental implications
In the present study, the adsorption of PHE by earthworms in-
dicates that earthworms might have a potential burden capacity for
HOCs, which is enlightening and might be significant in soil
pollution remediation. Eiseina foteida, as a model organism species
in ecotoxicological tests, has been questioned because some au-
thors found that they can tolerate high concentrations of
 Z. Shi et al. / Journal of Environmental Management 212 (2018) 115e120
contaminants in soil environments (Contreras-Ramos et al., 2006;
Tejada et al., 2011). Therefore, the use of Eiseina foteida in soil
pollution remediation has also been studied and acknowledged
(Contreras-Ramos et al., 2006; Hickman and Reid, 2008). Vermir-
emediation refers to the use of earthworms for removing con-
taminants from the soil or when earthworms help to degrade non-
recyclable chemicals (Rodriguez-Campos et al., 2014; Rorat et al.,
2017). Some terms have been used to describe the strategies of
vermiremediation under different scenarios such as ‘vermiremoval’
(Herna ndez-Castellanos et al., 2013) and ‘vermistimulation’ (Azizi
et al., 2013; Pan et al., 2016; Suthar, 2009). However, it seems
that little research has focused on the roles of ‘vermiaccumulation’
or ‘vermiextraction’ in vermiremediation of organic contaminants.
In the present study, the results obtained from the adsorption
experiments indicated that earthworms have the potential ability
to carry a large amount of HOCs. Given that HOCs concentration in
the soil is usually low, earthworms might be promising to be the
‘hyperaccumulators’ for HOCs. As a matter of fact, some studies
found that the concentration of HOCs in earthworms can be very
high, even when the concentration in soil was rather low (See
Supplementary Materials). The observation means that the feasi-
bility of using vermiaccumulation in vermiremediation of soils
contaminated with HOCs might at least be worth exploring.
Here, we attempt to provide a concise discussion about this
issue. There are at least three obstacles that need to be overcome to
fully validate the use of vermiaccumulation by and large: i) whether
earthworms can survive in polluted soil; ii) whether earthworms
can accumulate a large amount of contaminants; iii) whether
earthworms can be removed from contaminated soils. To solve the
first question, many studies have confirmed that earthworms might
survive in highly polluted soils (Rodriguez-Campos et al., 2014). For
the third question, many earthworm sampling methods, such as
chemical extraction methods (formalin expulsion, mustard water
suspension expulsion, ect.) and other expellants (electrical shock),
were useful for removing earthworms from polluted soils (Smith
et al., 2008; Valckx et al., 2011). However, the efficiency of these
methods stilled needed to be assessed in contaminated soils. For
the second question, the results in the present study indicate that
the partitioning of HOCs between the soil solution and earthworm
tissue is the key process limiting the efficiency of vermiaccumula-
tion. Therefore, through enhancing the bioavailability of HOCs, such
as with the aid of surfactants, this obstacle might be overcome (Liao
et al., 2015; White et al., 2007; Zhou et al., 2011).
Based on the above discussion, the use of vermiaccumulation in
vermiremediation might be feasible in some special cases. Since
there are many limitations, the use of vermiremediation for soil
pollution control should be carefully considered including the is-
sues of earthworm survival, accumulation capacity and the
bioavailability of pollutants.
Acknowledgments
Financial support from the Natural Science Foundation of Shanxi
Province (CN) (Project No. 201601D202089) and the Natural Sci-
ence Foundation of China (Grant No. 41701286) are highly
acknowledged.
Appendix A. Supplementary data
Supplementary data related to this article can be found at
https://doi.org/10.1016/j.jenvman.2018.01.079.
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