You are on page 1of 7

Mercury accumulation in marine bivalves: Inuences of biodynamics

and feeding niche


Ke Pan, Wen-Xiong Wang
*
Division of Life Science, The Hong Kong University of Science and Technology (HKUST), Clear Water Bay, Kowloon, Hong Kong
a r t i c l e i n f o
Article history:
Received 9 March 2011
Received in revised form
15 June 2011
Accepted 17 June 2011
Keywords:
Mercury
Methylmercury
Bivalves
Biodynamics
Subcellular distribution
Feeding niche
a b s t r a c t
Differences in the accumulation of mercury (Hg) in ve species of marine bivalves, including scallops
Chlamys nobilis, clams Ruditapes philippinarum, oysters Saccostrea cucullata, green mussels Perna viridis,
and black mussels Septifer virgatus, were investigated. The bivalves displayed different patterns of Hg
accumulation in terms of the body concentrations of methylmercury (MeHg) and total Hg (THg), as well
as the ratio of MeHg to THg. Parameters of the biodynamics of the accumulation of Hg(II) and MeHg
could reect the species-dependent Hg concentrations in the bivalves. With the exception of black
mussels, we found a signicant relationship between the efux rates of Hg(II) and the THg concentra-
tions in the bivalves. The interspecic variations in the MeHg to THg ratio were largely controlled by the
relative difference between the elimination rates of Hg(II) and MeHg. Stable isotope (d
13
C) analysis
indicated that the ve bivalve species had contrasting feeding niches, which may also affect the Hg
accumulation.
2011 Elsevier Ltd. All rights reserved.
1. Introduction
Bioaccumulation of mercury (Hg) has been a long-standing topic
since the mercury poisoning of thousands of people who consumed
Hg-contaminated sh and shellsh in Minamata Bay, Japan (Ditri,
1991). Hg is ubiquitous in natural waters, sediments, soils and air,
making it a global health concern. The organometallic form of Hg,
methylmercury (MeHg), is a potent neurotoxin and has a strong
tendency to be biomagnied in aquatic food chains, despite its low
environmental concentrations (Watras and Bloom, 1992; Ullrich
et al., 2001). One basic question surrounding Hg bioaccumulation
is how Hg concentrations of parts per trillion in water can yield Hg
concentrations of parts per million in sh (Morel et al., 1998; Chen
et al., 2008). Although considerable progress has been made in
understanding the Hg accumulation in aquatic organisms over the
past decades (Mason et al., 1995, 1996; Gagnon and Fisher, 1997;
Wang et al., 1998; Pan and Wang, 2004), there are still gaps in
our understanding of the biodynamics involved.
The accumulation of Hg in bivalve molluscs is of great interest
because they are widely consumed by humans worldwide. Bivalves
are strong metal accumulators and may act as an important vector
for pumping Hg to higher trophic levels. Previous studies have
indicated that bivalves can efciently absorb Hg(II) and MeHg from
water and sediments (Gagnon and Fisher, 1997; Pan and Wang,
2004), and environmental variables such as pH, temperature, dis-
solved organic carbon, and salinity have substantial effects on the
uptake of Hg (Inza et al., 1998; Tsui and Wang, 2004; Wang and
Wang, 2010). These studies are indeed valuable for interpreting
the crucial steps controlling the entry of Hg(II) and MeHg into
the food web. The interspecic divergence of Hg accumulation in
aquatic organisms is however less well known. Bivalve molluscs
reduce their metal toxicity through a series of mechanisms
including sequestration in metallothionein (MT), forming inert
granules by lysosomal vesicles for storage or exocytosis (Dallinger,
1993; Marigmez et al., 2002). The choice of detoxication strategy
affects the biodynamics of metals, leading to great variations in
metal concentrations among species.
Differences in the body concentrations of Hg in aquatic organ-
isms may reect the interspecic biodynamics of Hg, and may also
be caused by the choice of feeding habits by the animals. Food
partitioning is a common mechanism by which substantial differ-
ences in resource use occur between cohabiting species (Ross, 1986).
Bivalves constitute a diverse taxonomical group occupying a wide
habitat in coastal environments. These lter-feeders generally feed
opportunistically on any phytoplankton, detritus, or benthic algae
available in their habitats. Closely related species may feed on subtly
different food sources resulting in different accumulations of metals
(Rainbow, 1995). Our previous study has shown that changes in the
* Corresponding author.
E-mail address: wwang@ust.hk (W.-X. Wang).
Contents lists available at ScienceDirect
Environmental Pollution
j ournal homepage: www. el sevi er. com/ l ocat e/ envpol
0269-7491/$ e see front matter 2011 Elsevier Ltd. All rights reserved.
doi:10.1016/j.envpol.2011.06.029
Environmental Pollution 159 (2011) 2500e2506
growth environment can have signicant impact on food avail-
ability, quality and growth for scallops Chlamys nobilis, thus affecting
the related accumulation of Cd and Zn (Pan and Wang, 2008). Chen
et al. (2009) also demonstrated that concentrations of MeHg were
higher for pelagic-feeding sh than for benthic-feeding sh. These
studies imply that the feeding niche may have a great impact on
metal bioaccumulation, yet the link between feeding niche and the
bioaccumulation of Hg remains underexplored.
In this study, we therefore conducted a comprehensive investi-
gation on the inter-species difference in Hg(II) and MeHg bio-
accumulation in ve species of marine bivalves. We determined (1)
the difference in Hg bioaccumulation from water and food; (2) the
detoxication strategies and their links with the interspecic Hg
body concentrations; and (3) how different feeding niches affect Hg
accumulation. Our overall objective was to understand the inter-
species difference of Hg bioaccumulation in marine bivalves based
on the biodynamics of Hg and the feeding niche of the bivalves.
2. Materials and methods
2.1. Marine bivalves
Five species of marine bivalves commonly found along the east coasts of Hong
Kong were collected. Scallops Chlamys nobilis and green mussels Perna viridis were
collected from Dapeng Bay and Tolo Harbor, respectively. Clams Ruditapes philip-
pinarum, black mussels Septifer virgatus and rock oysters Saccostrea cucullata were
collected from Clear Water Bay. Individuals of a wide range of body sizes were
collected, which were acclimatized in circulating seawater (20

C and 33 psu) in
a 200 L water tank for one week. During the acclimation periods, the bivalves were
fed with diatoms Thalassiosira weissogii twice a day.
2.2. Total Hg and MeHg in seawater, sediments and bivalves
To measure the total Hg (THg), surface seawaters were sampled from Dapeng
Bay, Tolo Harbor and Clear Water Bay. The seawater samples were ltered with
Millipore 0.45 mm polycarbonate lters in a clean bench. Bromine monochloride
(BrCl) was added to ltered (for dissolved Hg) and unltered (for THg) samples to
produce a nal concentration of 0.5% (v/v). The samples were digested at room
temperature for 12 h prior to analysis by cold vapor atomic uorescence spec-
trometer (CVAFS) according to EPA method 1631E. Total suspended particulate
matter (SPM) was also determined by ltering an aliquot of seawater sample onto an
Advantec

GC-50 glass lter, which was rinsed with isotonic ammonium formate to
remove the salts. Surface sediments (1e2 cm, <63 mm, n 3) were collected from
Clear Water Bay.
For measurements of background THg and MeHg concentrations in the bivalves,
individuals of different sizes for each bivalve species were randomly selected from
the collection. Selected bivalves were depurated without feeding for two days.
Afterwards the bivalves were dissected, freeze-dried, and ground into a ne powder.
For small individuals (shell length <1 cm), 10e20 individuals of similar size were
pooled to make up a composite sample. For each site, totally 50 samples were made,
among which the number of composite samples was between 10 and 15. Dried
tissues or sediments were weighed and approximately 0.2 g of which were digested
at 190

C with aqua regia (2 mL HNO
3
:6 mL HCl) in a microwave digestion system.
MeHg in bivalves were extracted by digesting approximately 40 mg of homogenized
soft tissues with 25% KOH in methanol at 60

C for 3 h.
All containers for sampling and analysis of Hg were vigorously cleaned with hot
4 N HCl acid. THg was analyzed using the single gold trap amalgamation technique
by CVAFS (QuickTrace

8000, USA). Method detection limit (MDL) of THg was


0.18 ng L
1
. All bottle and lter blanks had THg belowthe MDL. Matrix recoveries for
seawater samples were 85e95%. MeHg was measured for biological samples with an
automated analytical system (MERX, Brooks Rand). Briey, the extract of bivalve
tissues was buffered with sodium acetate at pH 4.9, and ethylated by sodium tet-
raethylborate in a 40 mL Teon line borate glass bottle. The quantication of MeHg
was automatically carried out by the MeHg analyzer. Analytical accuracy of bio-
logical and sediment samples was checked by concurrent digestion and analysis of
certied reference material IAEA-142 (mussel homogenate) and NIST 1646a (estuary
sediments), with a recovery of 91e94% for THg and 89e96% for MeHg.
2.3. Biodynamics of Hg(II) and MeHg in bivalves
Bivalves of 20e30 mm shell height were used in the biodynamics experiments.
The radioisotope
203
Hg(II) (t
1/2
46.6 d, specic activity:162e200 GBq g
1
) used in
the study was purchased from Isotope Products Laboratories, Eckert & Ziegler,
Germany. Me
203
Hg was synthesized from the
203
Hg(II) following a well-established
method (Rouleau and Block, 1997). Parameters of the biodynamics of Hg(II) and
MeHg, the dissolved uptake constant, assimilation efciency, and excretion rate
constant, were then quantied.
To measure the inux rates of waterborne Hg(II) and MeHg, the radioisotope
203
Hg(II) or Me
203
Hg were added into the 0.22 mmltered seawater to give different
Hg concentrations (25, 50, 100, 200, 400 ng L
1
for Hg(II), and 10, 20, 50, 100,
200 ng L
1
for MeHg). Each concentration treatment contained four replicate
individuals, and the bivalves were placed individually in 500 mL medium held in
a Teon beaker. The exposure period was 30 min for Hg (II) and 5e10 min for MeHg,
during which the decline of metal concentration was within 20%. Afterwards the
bivalves were carefully rinsed with non-radioactive seawater and dissected, and
their radioactivities were determined. The whole animals were completely dried at
60

C to determine the dry weights. Hg(II) or MeHg inux rate (I


w
, mg g
1
day
1
) was
calculated as follows:
Iw
A
tissue
SA W t
;
where A
tissue
is the radioactivity in the whole soft tissue after exposure (ccpm), SA is
the specic activity of Hg(II) or MeHg in seawater (ccpm m g
1
), W is the dry weight
of the soft tissue (g), and t is the duration of exposure (d). The dissolved uptake rate
constant (k
u
) can be calculated by the inux rate divided by the metal concentration
in the dissolved phase (C
w
).
The assimilation efciencies (AEs) of Hg in marine diatoms T. weissogii and
sediments were determined using the pulse feeding and gamma radioactive tracer
method (Wang and Fisher, 1996; Pan and Wang, 2009a). Gamma detection is
nondestructive, thus the radioactivity in the same individual bivalve can be assessed
over time. The diatoms were grown in an f/2 mediumwith N, P, Si, vitamins and trace
metals minus Zn, Cu, and EDTA. A stock solution (500 mg L
1
) of sediments (<63 mm)
was prepared with 0.22 mm ltered seawater. The radioisotope
203
Hg(II) or Me
203
Hg
was added to exponentially growing diatoms at a concentration of 370 kBq L
1
and
the diatoms were grown for two days. The sediment was radiolabeled at 740 kBq L
1
for two days as well. The radiolabeled diatoms or sediments were collected by
centrifugation, and were washed in non-radioactive ltered seawater. There were 10
replicate individuals in each treatment. The bivalves were fed diatoms at a concen-
tration of 5 10
4
cells mL
1
or sediments at a concentration of 2 mg L
1
for 30 min.
After the pulse feeding, the bivalves were rinsed thoroughly with ltered seawater
and their initial radioactivities determined, and then placed separately into poly-
propylene beakers containing 200 mL of seawater held in a 10 L enclosed recirculating
ow-through aerated seawater aquarium. non-radioactive algae T. weissogii were
fed to the bivalves twice a day to depurate their guts of the ingested radiolabeled food.
Feces were removed frequently and the seawater was changed twice a day during the
experiment. Radioactivity in each bivalve was assayed at 12 h intervals over 48 h.
Assimilation efciency (AE) was determined as the percentage of the initial radio-
activity retained in bivalves after 48 h of depuration.
To measure the Hg(II) and MeHg efux rates of bivalves, fteen individuals of each
bivalve species were exposed to dissolved
203
Hg(II) or Me
203
Hg for 1 h per day (at
37 kBq L
1
). During the radioactive exposure, diatoms T. weissogii radiolabeled with
203
Hg(II) or Me
203
Hg were also fed to the bivalves. The dual-exposure lasted for six days
and all the bivalves were assayed for their initial activities on the seventh day. Two
bivalves were dissected, and the radioactivity in the tissues and shells was also counted.
The bivalves were depurated in non-radioactive seawater for 24 days. The radioactivity
in each bivalve as a whole animal was assayed periodically throughout the experiment.
Meanwhile, three individuals were sampled on the sixth day for subcellular metal
distribution analysis. The efux rate constant (k
e
, day
1
) was calculated from the slope
of the regression between the natural log of the percentage of
203
Hg(II) or Me
203
Hg
retained in the slow exchange compartment and the period of depuration.
2.4. Subcellular fractionation of Hg(II) and MeHg
The subcellular fractionation was carried out according to Wallace et al. (2003)
and ve fractions were separated (cellular debris, metal-rich granule fraction-MRG,
organelle, heat-sensitive protein [HSP], and metallothionein-like protein [MTLP]).
Briey, the soft tissues were homogenized in 5 mL of 30 mMTriseNaCl buffer (pH8.0;
0.15 M sodium chloride; 5 mM freshly prepared antiprotease, 2-mercaptoethanol;
0.1 mM phenylmethylsulfonyl uoride (PMSF)). The homogenate was centrifuged at
1450g at 4

C for 15 min. The separated pellet was the cellular debris fraction, which
was digested in 1 N NaOH at 80

C for 10 min, and centrifuged at 5000 g again for
10 min. The resulting pellet was the MRG and the supernatant was the cellular debris
fraction. The supernatant from the rst centrifugation was further subjected to
centrifugation at 100,000g at 4

C for 1 h. The pellet from this step was the organelle
fraction (organelles). After 80

C treatment of the supernatant fromthis step, the HSP


was denatured and separated by centrifugation at 50,000g and 4

C. The nal
supernatant was the MTLP fraction. Each fraction was assayed for radioactivity. The
metal subcellular distribution was dened as the percentage of
203
Hg(II) or Me
203
Hg
in each fraction.
2.5. Stable isotopic signature (d
13
C and d
15
N) for bivalves
Selected bivalves (shell length: w20 mm) were depurated in 0.22 mm ltered
seawater for two days to allow the evacuation of gut contents. The soft tissues were
K. Pan, W.-X. Wang / Environmental Pollution 159 (2011) 2500e2506 2501
dissected and freeze-dried, which were then ground into a ne powder. Five indi-
viduals were combined into one composite sample in order to obtain a representa-
tive sample (Herman et al., 2000). Stable isotopic measurements were performed
with Finnigan MAT Delta V advantage isotope ratio mass spectrometer. Results are
reported as comparisons with atmospheric nitrogen (for N) and Vienna Pee Dee
Belemnite (for C) as reference standards, and were calculated as: d
13
C or
d
15
N (R
sample
/R
standard
1) 1000, where R is the
13
C/
12
C or
15
N/
14
N ratio for d
13
C
or d
15
N, respectively. The analysis errors of reproducibility were within 0.2& for
both d
13
C and d
15
N.
2.6. Radioactivity measurements, data treatments and statistical analysis
The radioactivity of
203
Hg(II) and Me
203
Hg were determined using a wallac
1480 NaI(T1) gamma counter (Turku, Finland). All counts were related to
standards and spillover. The gamma emission of
203
Hg(II) and Me
203
Hg was
determined at 232 keV. Counting times were adjusted to yield a propagated
counting error of less than 5%. All counts data were corrected for decay before
any calculations. The statistical signicance of the regressions was tested by
analysis of variance, and the regressions were quantied by the coefcient of
determination, r
2
. Differences in AE and subcellular metal distribution among
the treatments were detected by one-way analysis of variance with a least-
signicant-difference (LSD) post hoc test in which all the percentage data were
arcsine transformed before analysis. Analysis of covariance (ANCOVA) was per-
formed to compare the THg and MeHg between different species of bivalves
(Fialkowski et al., 2003).
3. Results and discussion
3.1. Total Hg concentrations in seawater and sediments
Total dissolved Hg concentrations in the seawater were 0.20,
0.35, and 0.21 ng L
1
for Dapeng Bay, Tolo Harbor and Clear Water
Bay, respectively (Table 1). Total dissolved Hg accounted for about
50% of THg, which was 0.39 ng L
1
for Dapeng Bay, 0.64 ng L
1
for
Tolo Harbor, and 0.38 ng L
1
for Clear Water Bay. The calculated
THg concentrations in SPMwere 90, 97 and 106 ng g
1
for the three
sites. THg concentration in the sediments fromClear Water Bay was
only 65.8 3.0 ng g
1
(n 3), lower than that of sediments found
in SPM (p < 0.01). Generally, THg concentrations in the uncon-
taminated marine environments were in the range of
0.1e0.8 ng L
1
in seawater and in the range of 20e400 ng g
1
in
sediments (Ullrich et al., 2001). These data indicate that the three
studied sites were not affected by Hg contamination and the
bivalves were exposed to a similarly low level of Hg.
3.2. Interspecic accumulation of THg and MeHg
in bivalves from the led
THg and MeHg concentrations in each bivalve species are shown
in Table 1 and Fig. 1. Overall, the bivalves showed various accu-
mulation patterns for Hg in terms of concentration and the MeHg to
THg ratio. THg concentration was the highest in the black mussels
S. virgatus (67.7e165.1 ng g
1
, mean of 91.9 ng g
1
), followed by
oysters S. cucullata (34.3e106.4 ng g
1
, mean of 70.3 ng g
1
) and
scallops C. nobilis (39.8e99.5 ng g
1
, mean of 60.3 ng g
1
). Clams
R. philippinarum and green mussels P. viridis accumulated the least
THg, with an average concentration of 47.4 ng g
1
(32.4e108.5 ng g
1
) and 30.1 ng g
1
(24.7e42.9 ng g
1
), respec-
tively. In contrast, the total dissolved Hg concentration from their
sites of collection was the highest (0.35 ng L
1
, Tolo Harbor)
(Table 1). With regard to MeHg, the scallops had the highest
average MeHg concentration of 18.8 ng g
1
. Comparable MeHg
concentrations were observed for oysters and clams (15.2 ng g
1
and 16.6 ng g
1
, respectively), which were higher than in green
mussels and black mussels (8.5 ng g
1
and 10.4 ng g
1
respec-
tively). The effect of growth dilution on Hg accumulation was only
observed in scallops and green mussels (Fig. 1, Table 2). Interest-
ingly, the proportion of MeHg also varied among bivalve species.
For example, the black mussels typically had a lower MeHg to THg
ratio of 11% (Table 1), three times lower than that of clams (37%).
3.3. Biodynamics of Hg(II) and MeHg in the bivalves and its
relationship with subcellular distribution of Hg(II) and MeHg
The dissolved uptake rate of Hg(II) and MeHg increased linearly
with exposure concentration (Fig. 2). The k
u
(calculated by I
u
/C
w
) of
Hg(II) for the scallops was 32.8 L g
1
d
1
, approximately three times
that of the green mussels (11.4 L g
1
d
1
) and clams R. philippinarum
Table 1
Summary of THg and MeHg concentrations in seawater and bivalves collected fromthe studied sites. Data presented as mean SD. n 3 for THg, total dissolved Hg, and SPMin
seawater. n 50 for the body concentrations of THg and MeHg in bivalves. NA: not applicable.
Site Total Hg
(ng L
1
)
Total dissolved Hg
(ng L
1
)
SPM load
(mg L
1
)
Species Body burden of Hg
(ng g
1
)
MeHg/THg
(%)
Allometric function
(Y aW
b
)
THg MeHg THg MeHg
Dapeng Bay 0.39 0.05 0.21 0.04 2.0 0.3 Scallop C. nobilis 60.3 14.2 18.8 5.7 31.4 6.9 53.5W
0.13
17.0W
0.11
Tolo Harbor 0.65 0.10 0.35 0.01 3.1 0.2 Green mussel P. viridis 30.2 4.1 8.9 2.0 29.9 6.4 26.3W
0.10
NA
Clam R. philippinarum 47.4 15.3 16.6 3.5 37.6 11.8 NA NA
Clear Water Bay 0.38 0.02 0.20 0.01 1.7 0.1 Oyster S. cucullata 70.3 15.3 15.2 3.3 22.4 6. 8 NA NA
Black mussel S. virgatus 91.9 18.8 10.4 1.5 11.6 2.2 NA NA
Scallop
0 1 2 3 4
-
1
)
0
50
100
150
200
THg
MeHg
Green mussel
0.0 0.4 0.8 1.2 1.6
0
50
100
150
200
Black mussel
Tissue dry weight (g)
0.0 0.2 0.4 0.6 0.8 1.0
0
50
100
150
200
Clam
0.0 0.3 0.6 0.9 1.2
0
50
100
150
200
Oyster
Tissue dry weight (g)
0.0 0.2 0.4 0.6 0.8
C
o
n
c
e
n
t
r
a
t
i
o
n

(
n
g

g
-
1
)
C
o
n
c
e
n
t
r
a
t
i
o
n

(
n
g

g
0
50
100
150
200
Fig. 1. Concentrations of THg and MeHg (ng g
1
) relative to the body dry weights in
ve species of bivalves.
K. Pan, W.-X. Wang / Environmental Pollution 159 (2011) 2500e2506 2502
(8.9 L g
1
d
1
). Oysters S. cucullata and the black mussels had
comparatively lower k
u
s (4.4 and 3.5 L g
1
d
1
respectively).
Remarkably high k
u
s of MeHg were observed in the bivalves, which
were 10e20 times higher than those of Hg(II). The k
u
of MeHg was
the highest for the scallops (596 L g
1
d
1
), and the lowest for the
oysters (62.5 L g
1
d
1
). The absorption efciency (a
w
) was 3e8% for
Hg(II) and 44e79% for MeHg (Table 2).
The depuration patterns of radiotracers
203
Hg(II) and Me
203
Hg
after the pulse feeding in the ve bivalve species are shown in
Fig. 3. The radioactivity of
203
Hg adsorbed on the shells was less
than 5% of the overall accumulation. With diatoms as food, the
oysters and clams had relatively higher AEs of 46% and 47%
respectively, while the other bivalve species had an AE of around
30% for Hg(II). MeHg was more efciently absorbed from food than
Hg(II) regardless of the type of food. Nearly 80e90% of MeHg in
diatoms and 50e72% MeHg in sediments was assimilated by the
bivalves. The AEs of Hg(II) from sediments were only 5e20%. The
reported AEs of Hg(II) and MeHg for zooplankton were only around
15% and 60%, respectively; for a deposit-feeding polychaete Nereis
succinea the values were 7e28% and 66e84%, respectively (Mason
et al., 1995, 1996; Wang et al., 1998). The high k
u
s and AEs indi-
cate that the bivalves have a strong ability to absorb Hg from both
dissolved and dietary phases.
There were considerable differences in the loss rates of Hg(II) in
the bivalves (Fig. 4, Table 2). The green mussels had the highest
efux rate (0.060 day
1
), followed by the scallops (0.042 day
1
), the
clams (0.039 day
1
) and the black mussels (0.035 day
1
) (Table 2).
The oysters had the slowest efux rate for Hg(II) of 0.021 day
1
.
Depuration of MeHg was clearly slower than that of Hg(II) in
bivalves. Again, the oysters had the lowest efux rate for MeHg
(0.003 day
1
), while the green mussels and the black mussels had
relatively higher rates (0.006e0.007 day
1
). The biological half-life
(t
1/2
ln2/k
e
) for Hg(II) in bivalves was only 12e33 days, but was
99e231 days for MeHg.
The subcellular distributions of Hg(II) and MeHg in the ve
bivalve species are shown in Fig. 5. Both Hg(II) and MeHg were
mainly distributed in MTLP (30e60% for Hg(II) and 60% for MeHg).
MT can be a rather important protein for Hg, as both Hg(II) and
MeHg are characterized by an extremely high afnity for SH-
residues (Viarengo et al., 1994; Canesi et al., 1999). There was also
a major difference in the distributions of Hg(II) among the ve
bivalve species. The oysters typically stored over 60% of the accu-
mulated Hg(II) in the MTLP. This was obviously higher than in the
other four species. The scallops stored 40% of Hg(II) in this fraction
and the percentages for the other three species were similar (30%).
The green mussels had about 20% of Hg(II) distributed in the
organelles fraction, whereas the oysters had 9%. For MeHg, the
bivalves shared rather similar partitioning patterns, in which the
MeHg in MTLP fraction contributed to around 60% of the overall
MeHg and other fractions only made up a minor portion.
A signicant positive relationship was found between the efux
rate of Hg(II) and the percentage of Hg(II) distribution in the
organelles fraction, indicating that this subcellular fraction has
a close relationship with the Hg(II) elimination process (Fig. 6).
Besides being stored in MT, metals could be diverted from MT and
other cytosolic ligands to various forms of insoluble storage, where
the organelles lysosomes may play a signicant role (Fowler, 1978;
Marigmez et al., 2002). In bivalves, mussels in particular, the
abundant lysosomes for intracellular and extracellular digestion are
particularly important organelles for forming vesicle-bound gran-
ules which are further eliminated from the body by exocytosis
in digestive gland (Langston et al., 1998; Dimitriadis et al., 2003;
Table 2
Summary of biokinetics parameters in ve species of bivalves (Mean SD). k
u
: dissolved uptake constant; AE: assimilation efciency (n 10); k
e
: efux rate constant (n 10);
a
w
: absorption efciency; FR: ltration rate (from Pan and Wang, 2009a, b).
Species k
u
(L g
1
day
1
)
FR
(L g
1
h
1
)
a
w
(%) AE (%) k
e
(day
1
) Biological half-life
(t
1/2
ln2/k
e
, days)
Hg(II) MeHg Hg(II) MeHg Hg(II) MeHg Hg(II) MeHg Hg(II) MeHg
Scallop C. nobilis 32.8 595.8 36.4 9.4 3.8 68.2 15.5e28.4 50.1e86.3 0.042 0.007 0.005 0.003 16.5 138.6
Clam R. philippinarum 8.9 82.4 4.79 1.87 7.7 71.7 9.2e46.3 67.5e86.0 0.039 0.007 0.005 0.002 17.8 138.6
Oyster S. cucullata 4.4 62.5 3.29 1.76 5.5 79.2 18.4e48.2 70.5e91.9 0.021 0.005 0.003 0.001 33.0 231.0
Green mussel P. viridis 11.4 132.7 12.5 3.33 3.8 44.2 5.5e31.6 52.9e76.5 0.060 0.018 0.007 0.006 11.6 99.0
Black mussel S. virgatus 3.5 71.4 6.25 1.07 2.8 56.7 10.6e34.1 72.5e81.5 0.035 0.006 0.006 0.002 19.8 115.5
Hg(II)
Dissolved concentrations (g L
-1
)
0.01 0.1 1
I
n
f
l
u
x

r
a
t
e

(

g

g


d


)
-
1
-
1
0.01
0.1
1
10
100
1000
MeHg
0.01 0.1 1
0.01
0.1
1
10
100
1000
Scallop
Clam
Oyster
Green mussel
Black mussel
Fig. 2. Inux rates of Hg(II) and MeHg in ve species of bivalves at different ambient
concentrations. Data presented as mean SD (n 4). The relationships between metal
inux rate (I
w
, mg g
1
d
1
) and metal concentration in the dissolved phase (C
w
, mg L
1
)
in the bivalves are as follows (r
2
0.99, p < 0.001). For Hg(II), I
w
32.8C
w
1.02
for
scallops; I
w
8.9C
w
0.87
for clams; I
w
4.4C
w
1.05
for oyster; I
w
11.4C
w
1.06
for green
mussels; I
w
3.5C
w
1.29
for black mussels; For MeHg, I
w
595.8C
w
1.16
for scallops;
I
w
82.4C
w
1.13
for clams; I
w
62.5C
w
1.14
for oysters; I
w
132.7C
w
1.24
for green mussels;
I
w
71.4C
w
0.96
for black mussels.
MeHg-Sediment
Time (h)
0 10 20 30 40 50
1
10
100
Hg(II)-Algae
0 10 20 30 40 50
%

r
e
t
a
i
n
e
d
1
10
100
MeHg-Algae
0 10 20 30 40 50
1
10
100
Scallop
Clam
Oyster
Green mussel
Black mussel
Hg(II)-Sediment
Time (h)
0 10 20 30 40 50
%

r
e
t
a
i
n
e
d
1
10
100
Fig. 3. Retention of Hg(II) and MeHg in ve species of bivalves following a pulse
feeding of radiolabeled diatoms or sediments. Data presented as mean SD (n 10).
K. Pan, W.-X. Wang / Environmental Pollution 159 (2011) 2500e2506 2503
Domouhtsidou and Dimitriadis, 2000). The higher percentage of
Hg(II) distributed in the organelles fraction in the green mussels
implies that the green mussels transported the Hg(II) extracellu-
larly through organelles (possibly lysosomes) more actively,
leading to higher efux rates in this species.
3.4. Biodynamics in controlling Hg body concentrations and the
MeHg to THg ratio in bivalves
Metal bioaccumulation in aquatic invertebrates can be highly
variable, even for animals within closely related taxa down to
species in the same genus (Rainbow, 2002). In our study, the body
concentrations of THg and MeHg in the bivalves also varied in terms
of concentration as well as MeHg to THg ratio. The site-specic
dissolved Hg concentration in seawater did not control the inter-
specic difference. For example, relatively higher THg concentra-
tions (0.35 ng L
1
) were observed inTolo Harbor, compared to those
observed in Dapeng Bay and Clear Water Bay (0.21 and 0.20 ng L
1
,
respectively). The green mussels collected from Tolo Harbor
contrarily contained the lowest THg concentrations in the tissues
among the ve bivalve species. Moreover, the clams, oysters, and
black mussels, which were all collected from Clear Water Bay,
accumulated THg differently (black mussels > oysters > clams).
Thus the differentiation of Hg bioaccumulation may be more likely
due to the biologically driven species-specic biodynamics. It
should be noted, however, that the concentrations of Hg(II) both in
water and in food were much higher than those of MeHg, and also
accounted for 60e90% of THg in the tissues of the bivalves. It is
possible that the biodynamics of Hg(II) had larger effects on the
concentrations of THg in the bivalves than that of MeHg.
The bivalves had contrasting k
u
for both Hg(II) and MeHg, which
varied by an order of magnitude (Table 2). Food quality also had
large effects on the AEs of Hg(II) and MeHg, indicating that food
choice may contribute to the species-specic Hg bioaccumulation
(see below). No signicant relationship can be found between the
k
u
or AE and the concentrations of THg and MeHg in the bivalves.
Not surprisingly, none of the parameters alone can explain the
interspecic body concentration of MeHg and THg, as bio-
accumulation is a result of the interaction between uptake and loss.
Yet, the biodynamics still has signicant implications for under-
standing the variations in the concentrations of MeHg and THg in
the bivalves. For example, the scallops had the highest k
u
both for
Hg(II) and MeHg, which may be associated with its highest MeHg
concentration. However, the high loss rate of Hg(II) (0.04 day
1
)
prevented the THg in scallops from ranking the highest.
Metal excretion rate constant (k
e
) was important for deter-
mining the intraspecic and interspecic differences in metal bio-
accumulation (Pan and Wang, 2009a, b). Oysters had the lowest k
e
of Hg(II) and MeHg in the bivalves (Table 2, Fig. 4). The k
e
of Hg(II) in
green mussels (0.06 day
1
) was two times higher than that in
oysters (0.02 day
1
). Similarly, the rate of MeHg in the green
mussels (0.007 day
1
) also overwhelmed that in the oysters (0.003
day
1
). Despite the fact that both the dissolved THg in Tolo Harbor
and the k
u
in green mussels were high, the fast loss rates of Hg(II)
and MeHg were partially responsible for the lowest THg and MeHg
concentrations in this species. A previous study has shown that the
Cu body concentrations in different bivalves were inversely corre-
lated with the Cu k
e
s (Pan and Wang, 2009a). Such a relationship
was also observed between the k
e
s of Hg(II) and the THg concen-
trations in the oysters, clams, scallops, green mussels (data not
shown). One exception was the black mussels. The k
e
s of Hg(II) and
MeHg (0.035 and 0.006 day
1
, respectively) were signicantly
higher than those in the oysters collected from the same site.
Contrarily, the black mussels contained higher THg concentrations
(92 ng g
1
) than the oysters (70 ng g
1
).
Growth dilution may also play a signicant role in controlling
the interspecic variation. Rapid growth is the major factor
resulting in reduced Hg concentrations in sh (Ward et al., 2010).
A specic growth rate ranging from 0.01 to 0.10 day
1
is commonly
MeHg
0 5 10 15 20 25
10
100
Hg(II)
Time (days)
0 5 10 15 20 25
%

r
e
t
a
i
n
e
d
10
100
Scallop
Clam
Oyster
Green mussel
Black mussel
Time (days)
Fig. 4. Depuration of Hg(II) and MeHg in ve species of bivalves over a 24-day period
following a 6-day combined exposure to dissolved and dietary Hg (Hg(II) or MeHg,
Mean SD, n 10).
S
c
a
l
l
o
p

C
l
a
m

O
y
s
t
e
r

B

m
u
s
s
e
l

G

m
u
s
s
e
l

S
c
a
l
l
o
p

C
l
a
m

O
y
s
t
e
r

B

m
u
s
s
e
l

G

m
u
s
s
e
l

0
20
40
60
80
MRG
Cellular debris
Organelles
HSP
MTLP
Hg(II)
%

m
e
t
a
l

i
n

e
a
c
h

f
r
a
c
t
i
o
n

0
20
40
60
80
MeHg
Fig. 5. Subcellular distribution of Hg(II) and MeHg in ve species of bivalves collected on the sixth day during a 24-day depuration period. (Mean SD, n 3). G mussel: green
mussel; B mussel: black mussel.
Hg(II) in organelles fraction (%)
0 5 10 15 20 25
E
f
f
l
u
x

r
a
t
e

(
d
a
y


)

-
1

0.00
0.02
0.04
0.06
0.08
0.10
Hg(II) in MTLP fraction (%)
0 20 40 60 80 100
0.00
0.02
0.04
0.06
0.08
0.10
y =0.0041 x -0.013
p<0.001
r
2
=0.96
Fig. 6. Relationship between the efux rate constant of ve species of bivalves and the
percentage of Hg(II) in either the organelles or the MTLP fraction. (n 10 for efux rate
and n 3 for subcellular distribution).
K. Pan, W.-X. Wang / Environmental Pollution 159 (2011) 2500e2506 2504
observed for marine bivalves (Clausen and Riisgrd, 1996;
Jrgensen, 1996). Thus, growth dilution can contribute to 10e30%
of the overall efux for Hg(II) when the specic growth rate
constant (g) is 0.01 per day, but can be up to 60e80% when the g is
as high as 0.10 per day. It is possible that the ve bivalve species
have different g and thus have different degrees of dilution for THg
and MeHg. However, the effects of growth may be complicated by
the rate of weight change compared to the rate of uptake and loss
for Hg, which may be the reason that the effect of growth dilution
on Hg accumulation was only observed in scallops and green
mussels (Fig. 1).
Biodynamics not only controls the body concentrations of Hg(II)
and MeHg, but also has confounding effects on the MeHg to THg
ratios. Bivalves effectively absorbed Hg(II) and MeHg from both
dissolved and dietary phases, with MeHg being accumulated more
efciently. Consequently, the MeHg to THg ratios in bivalves were
signicantly elevated compared to those in water and sediments
(less than 0.5%, Ullrich et al., 2001). Although MeHg concentrations
in bivalves did not vary widely (a difference of w2 times), there was
a difference of nearly four times (11e37%) in the MeHg to THg ratio
(Table 1). On the one hand, the relatively low percentage of Hg in
the form of MeHg in bivalves was largely determined by the much
higher concentrations of Hg(II) in water and food as compared to
MeHg. On the other hand, the difference in the MeHg to THg ratio
was affected by the differences between the biodynamics of Hg(II)
and MeHg in each species. However, it appeared that the k
u
and AE
could not explain the species-dependent ratio. For example,
difference in k
u
between Hg(II) and MeHg was most pronounced in
the black mussel (a difference of w20 times), whereas the ratio was
the lowest in this species. The proportion of MeHg was positively
correlated with the ratio of the efux rate of Hg(II) to that of MeHg
(Fig. 7, p 0.059), indicating that the species-specic MeHg to THg
ratio was possibly controlled by the relative difference between the
efux rate of Hg(II) and MeHg in each species. The ratio of the efux
rate of Hg(II) to that of MeHg in bivalves was found to be 6e9.
A similar difference (3e7 times) has also been observed in sh
(Wang and Wong, 2003; Wang et al., 2010). However, the
predominance of dietary exposure and ingestion of food of a higher
MeHg to THg ratio made the proportion of MeHg in high-trophic-
level shes overwhelm that in bivalves.
3.5. Feeding niche of bivalves and its effects on Hg accumulation
While the species-specic accumulation of Hg was driven by the
detoxication strategies of bivalves, the impacts of ecological
factors cannot be ruled out. Field studies have shown an average
15
Nenrichment of 3.4&per trophic level, and a lower value of 2.5&
was observed for herbivores (Vander Zanden and Rasmussen,
2001). The d
15
N only varied within a small range of 1& between
the ve bivalve species (Table 3), implying a similar trophic level
between the bivalves. There were however signicant differences
in d
13
C signals among the ve species of bivalves (Table 3), indi-
cating that they may rely on different carbon sources. The d
13
C
signals varied largely between clams (17.1&) and black mussels
(19.5&) collected from Clear Water Bay, with an intermediate
value of 18.8& for oysters. Numerous studies have shown that
phytoplankton or SPM are more depleted in d
13
C than sedimentary
organic matter (SOM) or benthic microalgae (France, 1995;
Middelburg and Nieuwenhuize, 1998; Ishihi, 2003), resulting in
pelagic feeders that are more depleted in d
13
C than benthic
consumers (Hobson et al., 1995). For example, Currin et al. (1995)
suggested that an average d
13
C of 14.9& was found for benthic
microalgae and an average of 21.1&for marine phytoplankton. In
our study, the clams appeared to feed on resuspended sediments,
whereas the black mussels relied onphytoplankton or SPMinwater
column. The rock oysters possibly relied on both sediments and
SPM. The different food sources and habitats may lead to different
bioavailability of Hg for clams, oysters and black mussels. It was
found that the THg concentration in the SPM (w100 ng g
1
) was
higher than that in the sediments (w65 ng g
1
), while the AEs of
Hg(II) and MeHg were both lower for sediments than phyto-
plankton. The black mussels may have a higher potential for
accumulating Hg from food than the clams and oysters. As a result,
the concentration of THg in black mussels was much higher than
that in oysters even though the oysters had the lowest elimination
rates of both Hg(II) and MeHg.
4. Conclusions
Our study has clearly demonstrated that bivalves have
substantial capacity to accumulate Hg(II) and MeHg fromwater and
food. All ve bivalve species, scallops, clams, oysters, green mussels
and black mussels, signicantly accumulated Hg from the low Hg
environments. The bivalves showed distinct detoxication strate-
gies for Hg, which was reected in their subcellular partitioning
and biodynamics. Hg(II) distributing in the organelles fraction was
closely related to the species-dependent Hg(II) elimination rate,
suggesting an interaction between the subcellular partitioning and
biokinetics of Hg(II) in bivalves. The biodynamics parameters were
only able to partially explain the different body concentrations of
THg and MeHg in bivalves, and the variations in the MeHg to THg
ratios were largely controlled by the relative difference between
the elimination rates of Hg(II) and MeHg. The interspecic body
concentrations of MeHg and THg were complicated by the differ-
ences in the biodynamics of Hg(II) and MeHg, and the different
feeding niches of bivalves.
Acknowledgements
We thank Prof. Ling-Feng Huang and his student Mr. Xin-Qing
Zheng of Xiamen University for their help in analyzing stable
isotopes in this study. This study was supported by the General
Research Funds from the Hong Kong Research Grants Council
(663009 and 662610).
k
e(Hg(II))
/k
e(MeHg)
5 6 7 8 9 10
M
e
H
g
/
T
H
g

r
a
t
i
o

(
%
)
0
10
20
30
40
50
y=7.84x-32.4
p=0.059
r
2
=0.74
Fig. 7. Relationship between the ratio of MeHg to THg and the ratio of the elimination
rates of Hg(II) and MeHg.
Table 3
The values of d
13
C and d
15
N in ve species of bivalves (mean SD, n 3).
Species Collection site d
13
C (&) d
15
N (&)
Scallop C. nobilis Dapeng Bay 18.7 0.1 9.0 0.1
Green mussel P. viridis Tolo Harbor 20.7 0.0 8.6 0.2
Clam R. philippinarum Clear Water Bay 17.1 0.0 8.3 0.2
Oyster S. cucullata Clear Water Bay 18.8 0.0 8.5 0.1
Black mussel S. virgatus Clear Water Bay 19.5 0.0 8.0 0.2
K. Pan, W.-X. Wang / Environmental Pollution 159 (2011) 2500e2506 2505
References
Canesi, L., Viarengo, A., Leonzio, C., Filippelli, M., Gallo, G., 1999. Heavy metals and
glutathione metabolism in mussel tissues. Aquatic Toxicology 46, 67e76.
Chen, C.Y., Serrell, N., Evers, D.C., Fleishman, B.J., Lambert, K.F., Weiss, J., Mason, R.P.,
Bank, M.S., 2008. Meeting report: methylmercury in marine ecosystems-from
sources to seafood consumers. Environmental Health Perspectives 116,
1706e1712.
Chen, C.Y., Dionne, M., Mayes, B.M., Ward, D.M., Sturup, S., Jackson, B.P., 2009.
Mercury bioavailability and bioaccumulation in estuarine food webs in the gulf
of Maine. Environmental Science and Technology 43, 1804e1810.
Clausen, I., Riisgrd, H.U., 1996. Growth, ltration and respiration in the mussel
Mytilus edulis: no evidence for physiological regulation of the lter-pump to
nutritional needs. Marine Ecology Progress Series 141, 37e45.
Currin, C.A., Newell, S.Y., Paerl, H.W., 1995. The role of standing dead spartina-
alterniora and benthic microalgae in salt-marsh food webs e considerations
based on multiple stable-isotope analysis. Marine Ecology Progress Series 121,
99e116.
Dallinger, R., 1993. Strategies of metal detoxication in terrestrial invertebrates. In:
Dallinger, R., Rainbow, P.S. (Eds.), Ecotoxicology of Metals in Invertebrates.
Lewis Publisher, Boca Raton, USA, pp. 245e289.
Dimitriadis, V.K., Domouhtsidou, G.P., Raftopoulou, E., 2003. Localization of Hg and
Pb in the palps, the digestive gland and the gills in Mytilus galloprovincialis (L.)
using autometallography and X-ray microanalysis. Environmental Pollution
125, 345e353.
Ditri, F.M., 1991. Mercury contamination e what we have learned since Minamata.
Environmental Monitoring and Assessment 19, 165e182.
Domouhtsidou, G.P., Dimitriadis, V.K., 2000. Ultrastructural localization of heavy
metals (Hg, Ag, Pb, and Cu) in gills and digestive gland of mussels, Mytilus
galloprovincialis (L.). Archives of Environmental Contamination and Toxicology
38, 472e478.
Fialkowski, W., Rainbow, P.S., Smith, B.D., Zmudzinski, L., 2003. Seasonal variation in
trace metal concentrations in three talitrid amphipods from the gulf of Gdansk,
Poland. Journal of Experimental Marine Biology and Ecology 288, 81e93.
Fowler, B.A., 1978. General subcellular effects of lead, mercury, cadmium, and
arsenic. Environmental Health Perspectives 22, 37e41.
France, R.L., 1995. C-13 enrichment in benthic compared to planktonic algae e
foodweb implications. Marine Ecology Progress Series 124, 307e312.
Gagnon, C., Fisher, N.S., 1997. Bioavailability of sediment-bound methyl and inor-
ganic mercury to a marine bivalve. Environmental Science and Technology 31,
993e998.
Herman, P.M.J., Middelburg, J.J., Widdows, J., Lucas, C.H., Heip, C.H.R., 2000. Stable
isotopes as trophic tracers: combining eld sampling and manipulative label-
ling of food resources for macrobenthos. Marine Ecology Progress Series 204,
79e92.
Hobson, K.A., Ambrose, W.G., Renaud, P.E., 1995. Sources of primary production,
benthic-pelagic coupling, and trophic relationships within the Northeast water
Polynya: insights from delta C-13 and delta N-15 analysis. Marine Ecology
Progress Series 128, 1e10.
Inza, B., Ribeyre, F., Boudou, A., 1998. Dynamics of cadmium and mercury
compounds (inorganic mercury or methylmercury): uptake and depuration in
Corbicula uminea. Effects of temperature and pH. Aquatic Toxicology 43,
273e285.
Ishihi, Y., 2003. Feeding of the bivalve Theora lubrica on benthic microalgae: isotopic
evidence. Marine Ecology Progress Series 255, 303e309.
Jrgensen, C.B., 1996. Bivalve lter feeding revisited. Marine Ecology Progress Series
142, 287e302.
Langston, W.J., Bebianno, M.J., Burt, G.R., 1998. Metal handling strategies in
molluscs. In: Langston, W.J., Bebianno, M.J. (Eds.), Metal Metabolism in Aquatic
Environments. Chapman & Hall, London, pp. 219e283.
Marigmez, I., Soto, M., Cajaraville, M.P., Angulo, E., Giamberini, L., 2002. Cellular
and subcellular distribution of metals in molluscs. Microscopy Research and
Technique 56, 358e392.
Mason, R.P., Reinfelder, J.R., Morel, F.M.M., 1995. Bioaccumulation of mercury and
methylmercury. Water Air and Soil Pollution 80, 915e921.
Mason, R.P., Reinfelder, J.R., Morel, F.M.M., 1996. Uptake, toxicity, and trophic
transfer of mercury in a coastal diatom. Environmental Science and Technology
30, 1835e1845.
Middelburg, J.J., Nieuwenhuize, J., 1998. Carbon and nitrogen stable isotopes in
suspended matter and sediments from the Schelde estuary. Marine Chemistry
60, 217e225.
Morel, F.M.M., Kraepiel, A.M.L., Amyot, M., 1998. The chemical cycle and bio-
accumulation of mercury. Annual Review of Ecology and Systematics 29,
543e566.
Pan, J.F., Wang, W.-X., 2004. Uptake of Hg(II) and methylmercury by the green
mussel Perna viridis under different organic carbon conditions. Marine Ecology
Progress Series 276, 125e136.
Pan, K., Wang, W.-X., 2008. Validation of biokinetic model of metals in the scallop
Chlamys nobilis in complex eld environments. Environmental Science and
Technology 42, 6285e6290.
Pan, K., Wang, W.-X., 2009a. Biodynamics to explain the difference of copper body
concentrations in ve marine bivalve species. Environmental Science and
Technology 43, 2137e2143.
Pan, K., Wang, W.-X., 2009b. Inter-individual variations in cadmium and zinc
biodynamics in the scallop Chlamys nobilis. Marine Ecology Progress Series 383,
151e160.
Rainbow, P.S., 1995. Biomonitoring of heavy metal availability in the marine envi-
ronment. Marine Pollution Bulletin 31, 183e192.
Rainbow, P.S., 2002. Trace metal concentrations in aquatic invertebrates: why and
so what? Environmental Pollution 120, 497e507.
Ross, S.T., 1986. Resource partitioning in sh assemblages e a review of eld
studies. Copeia, 352e388.
Rouleau, C., Block, M., 1997. Working methods paper e fast and high-yield synthesis of
radioactive (CH
3
Hg)-Hg-203(II). Applied Organometallic Chemistry 11, 751e753.
Tsui, M.T.K., Wang, W.-X., 2004. Temperature inuences on the accumulation and
elimination of mercury in a freshwater cladoceran, Daphnia magna. Aquatic
Toxicology 70, 245e256.
Ullrich, S.M., Tanton, T.W., Abdrashitova, S.A., 2001. Mercury in the aquatic envi-
ronment: a review of factors affecting methylation. Critical Reviews in Envi-
ronmental Science and Technology 31, 241e293.
Vander Zanden, M.J., Rasmussen, J.B., 2001. Variation in delta N-15 and delta C-13
trophic fractionation: implications for aquatic food web studies. Limnology and
Oceanography 46, 2061e2066.
Viarengo, A., Canesi, L., Moore, M.N., Orunesu, M., 1994. Effects of Hg
2
and Cu
2
on
the cytosolic Ca
2
level in molluscan blood-cells evaluated by confocal
microscopy and spectrouorometry. Marine Biology 119, 557e564.
Wallace, W.G., Lee, B.G., Luoma, S.N., 2003. Subcellular compartmentalization of Cd
and Zn in two bivalves. I. Signicance of metal-sensitive fractions (MSF) and
biologically detoxied metal (BDM). Marine Ecology Progress Series 249,
183e197.
Wang, W.-X., Fisher, N.S., 1996. Assimilation of trace elements and carbon by the
mussel Mytilus edulis: effects of food composition. Limnology and Oceanog-
raphy 41, 197e207.
Wang, R., Wang, W.-X., 2010. Importance of speciation in understanding mercury
bioaccumulation in tilapia controlled by salinity and dissolved organic matter.
Environmental Science and Technology 44, 7964e7969.
Wang, W.-X., Wong, R.S.K., 2003. Bioaccumulation kinetics and exposure pathways
of inorganic mercury and methylmercury in a marine sh, the sweetlips Plec-
torhinchus gibbosus. Marine Ecology Progress Series 261, 257e268.
Wang, W.-X., Stupakoff, I., Gagnon, C., Fisher, N.S., 1998. Bioavailability of inorganic
and methylmercury to a marine deposit feeding polychaete. Environmental
Science and Technology 32, 2564e2571.
Wang, R., Wong, M.H., Wang, W.-X., 2010. Mercury exposure in the freshwater
tilapia Oreochromis niloticus. Environmental Pollution 158, 2694e2701.
Ward, D.M., Nislow, K.H., Chen, C.Y., Folt, C.L., 2010. Rapid, efcient growth reduces
mercury concentrations in stream-dwelling Atlantic salmon. Transactions of the
American Fisheries Society 139, 1e10.
Watras, C.J., Bloom, N.S., 1992. Mercury and methylmercury in individual
zooplankton e implications for bioaccumulation. Limnology and Oceanography
37, 1313e1318.
K. Pan, W.-X. Wang / Environmental Pollution 159 (2011) 2500e2506 2506