Science of the Total Environment 333 (2004) 149 165

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Contaminant levels in sediments and asteroids (Asterias rubens

L., Echinodermata) from the Belgian coast and Scheldt estuary:
polychlorinated biphenyls and heavy metals
B. Danis a,*, P. Wantier b, S. Dutrieux a, R. Flammang b, Ph. Dubois a, M. Warnau c
a
Laboratoire de Biologie Marine (CP 160/15), Universite Libre de Bruxelles, 50 avenue F.D. Roosevelt, B-1050 Brussels, Belgium
b
Laboratoire de Chimie Organique, Universite de Mons-Hainaut, 19 avenue Maistriau, B-7000 Mons, Belgium
c
International Atomic Energy Agency-Marine Environment Laboratory, 4 Quai Antoine Ier, MC-98000 Monaco
Received 26 November 2003; received in revised form 29 April 2004; accepted 5 May 2004

Abstract

The Southern Bight of the North Sea is particularly exposed to anthropogenic contamination, due to heavy urbanisation and
industrialisation of its catchment area. The present work focuses on polychlorinated biphenyl (PCB) and metal contamination of
the marine environment along and off the Belgian coast. Its objectives were to compare the concentrations of seven PCB
congeners and four heavy metals in the sediments (a repository for anthropogenic contaminants) and in the asteroid Asterias
rubens (a recognized bioindicator species). Nineteen sampling stations were considered between the mouth of the Scheldt
Estuary and the southern limit of the Belgian coast (asteroids were found in 10 out of the 19 stations). PCB and metal
concentrations measured in sediments and asteroids were in the range of values reported in previous studies. Stations under
direct influence of the Scheldt were the most impacted by the considered contaminants. Metal concentrations varied according
to the grain-size fraction considered. In asteroids, PCBs and metals were found to be selectively distributed among body
compartments, and pyloric caeca were found to most efficiently discriminate between sampling stations contamination levels.
PCB and metal analysis of sediments provided a physicochemical evaluation of the contamination, whereas analysis of asteroids
introduced a biological dimension to the approach by taking into account bioavailability of the contaminants.
D 2004 Elsevier B.V. All rights reserved.

Keywords: Echinoderms; Asterias rubens; Heavy metals; Polychlorinated biphenyls; Sediments; North Sea

1. Introduction nant fluxes to the sea) running through its catchment

The Southern Bight of the North Sea is a highly
urbanized and industrialized region, with important
rivers (viz. major vectors of anthropogenic contami-
* Corresponding author. Tel.: +32-2-6502970; fax: +32-2-
6502796.
E-mail address: bdanis@ulb.ac.be (B. Danis).
area (NSTF, 1993a,b). These rivers carry considerable
amounts of contaminants of domestic, industrial and
agricultural origins (NSTF, 1993a,b; Bayens, 1998)
which eventually reach the marine environment.
When reaching marine waters, contaminants main-
ly concentrate in the sediments due to their generally
low solubility in sea water and their tendency to
adsorb onto particles. Sediments therefore constitute
the bulk of anthropogenic contaminants in the coastal

0048-9697/$ - see front matter D 2004 Elsevier B.V. All rights reserved.
doi:10.1016/j.scitotenv.2004.05.002
150 B. Danis et al. / Science of the Total Environment 333 (2004) 149165

environment and may be a major source of contam-
ination for numerous organisms living in or close to
them (Stebbing et al., 1992; NSTF, 1993a; Karbe et
al., 1994; Alzieu and Michel, 1998). Among the latter
organisms, the common asteroid Asterias rubens is a
widely distributed and abundant top-predator species
in the North Sea which is known to influence the
structure of benthic communities (Menge, 1982; Hay-
ward and Ryland, 1990; Hostens and Hammerlinck,
1994). This species has proved to be an efficient tool
for biomonitoring of several anthropogenic contami-
nants (e.g., PCBs, metals, organometals) in laboratory
and/or field studies (e.g., Bjerregaard, 1988; Everaarts
and Fischer, 1989; Temara et al., 1997, 1998a,b;
Warnau et al., 1999).
The present study is part of the Sustainable
Development of the North Sea Program of the SSTC
(Belgian Government Prime Minister Services) and
focuses on anthropogenic contamination of the Bel-
gian marine environment. This marine region is di-
rectly exposed to large amounts of anthropogenic
contaminant carried by the Scheldt river which ends
up between the northern limit of the Belgian coast and
the southern limit of the Dutch coast (see, e.g.,
Bayens, 1998). Among these contaminants, poly-
chlorinated biphenyls (PCBs) and metals are of par-
ticular concern (NSTF, 1993a; Mommaerts et al.,
1994; Alzieu and Michel, 1998; Laane et al., 1999).
The aim of the present paper was to determine the
concentrations of seven PCB congeners (IUPAC# 28,
52, 101, 118, 138, 153, 180) and four heavy metals
(Zn, Cd, Cu, and Pb) in sediments and in the asteroid
A. rubens in order to (i) compare and integrate the
information gathered through these two indicators and
(ii) assess the quality status of the considered region.
2. Materials and methods
2.1. Sampling and sample preparation
Samples (sediment and asteroids) were collected
between February 4th and March 31st 1998 during a
cruise with the RV Belgica and by seashore fishing
and scuba diving. Nineteen sampling stations were
considered (Table 1, Fig. 1); 18 of them are located
along the coast or in the open sea (these stations may
be arranged according to seven transects parallel or
perpendicular to the coast) and one is located in a
closed estuary (Scharendijke, Grevelingenmeer, The
Netherlands). Sediments (six replicates per station)
were collected either using a Reineck box-corer dur-

Table 1
Positions and characteristics of the sampling stations
Station code Coordinates Depth Salinity Date of sampling Sediment Asteroid
(N) (O) (m) (p.s.u.) (mm/dd/yy) collection collection
120 51j11.10 2j42.07 11.6 33.4 02/05/98 yes no
130 51j16.25 2j54.30 8.32 32.0 02/05/98 yes no
140 51j20.00 3j24.00 5.73 31.2 02/05/98 yes no
150 51j25.00 3j24.00 13.7 32.1 02/05/98 yes no
230 51j18.50 2j51.00 11.5 32.2 02/13/98 yes yes
250 51j31.00 3j19.00 9.65 32.0 02/05/98 yes yes
330 51j26.00 2j48.50 24.0 33.4 02/06/98 yes yes
700 51j22.60 3j13.20 10.9 31.1 02/11/98 yes no
710 51j26.45 3j08.32 12.2 32.6 02/11/98 yes yes
Breskens 51j24.40 3j30.00 31.0 03/05/98 yes yes
Knokke 51j20.80 3j17.80 32.0 03/20/98 yes yes
Nieuwpoort 51j08.80 2j42.80 32.5 03/06/98 yes yes
Oostende 51j13.80 2j54.40 32.0 03/10/98 yes yes
Perkpolder 51j24.80 4j02.60 27.0 03/09/98 yes no
S01 51j25.00 3j34.20 17.0 29.9 02/05/98 yes no
Terneuzen 51j20.70 3j48.30 28.0 03/09/98 yes no
Wenduine 51j17.80 3j04.40 32.7 03/19/98 yes yes
ZG03 51j15.70 2j40.00 18.6 32.6 02/11/98 yes no
Scharendijke 51j44.50 3j50.70 5 15 29.0 03/04/98 yes yes
 B. Danis et al. / Science of the Total Environment 333 (2004) 149165 151

Fig. 1. Sampling stations and transects along and off the Belgian coast.

ing the cruise or by hand in the intertidal zone during ately frozen ( 20 jC) in 400 ml acid-washed poly-
seashore collection. In both cases, only the upper 5 cm propylene containers. Asteroids (n=15 per station)
layer of the sediments was considered and immedi- were collected either using a beam trawl (during the
152 B. Danis et al. / Science of the Total Environment 333 (2004) 149165
cruise; trawling time ranged from 10 to 30 min,
depending on the sampling conditions, i.e., currents,
bottom nature, etc.) or by hand (seashore fishing) or
scuba diving. Only specimens belonging to the same
size class were considered (5 7 cm from the tip of
the arm to the mouth). Immediately after collection,
asteroids were dissected into five compartments (oral
body wall, aboral body wall, gonads, gut and pyloric
caeca). Dissected body compartments were pooled by
three (leading thus to five pools per station, each pool
coming from three individuals) and frozen at 20 jC.
Tissues to be analyzed for PCB content (all except
gut, due to too little tissue quantities) were wrapped in
aluminum foils immediately after dissection and fro-
zen at 20 jC until analysis. Prior to metal analyses,
sediment and asteroid samples were dried at 80 jC for
72 h. In addition, dried sediments were sieved using
an Endicots Octagon 200 siever in order to determine
grain-size distributions. The different fractions (>4,
2 4, 1 2 mm, 500 1000, 250 500, 125 250, 63
125, and<63 Am) were collected and weighed. Frac-
tions larger than 1000 Am most generally accounted
for less than 2% of total sediment weight; therefore,
only the five finest fractions were considered (indi-
vidually) for subsequent metal analyses. PCB analy-
ses were performed on the bulk sediment alone.
2.2. PCB analyses
All solvents and reagents were pesticide grade.
PCB congeners and reference material were bought
from Promochem (Germany).
Part of the frozen sample was taken, weighed and
dried overnight at 103 jC to determine the dry weight
and water content. The sample was homogenized and
a surrogate (PCB#103) was added. Samples were then
extracted three times with a modified Bligh and Dyer
method (methanol, dichloromethane and water;
MeOH:CH2Cl2:H2O, 10:5:4) (Booij and Van den
Berg, 1994) and added with an NaCl solution. The
organic phase was separated and the aqueous phase
was extracted three times with dichloromethane.
Extracts were then gathered. A fraction of the extracts
was used to determine the total lipid content (gravi-
metrically). The remaining part was evaporated under
nitrogen and 3 ml of isooctane were added to each
sample. Lipids were eliminated by precipitation with
sulfuric acid (three times). For sediment analysis, an
additional purification step using mercury to precipi-
tate sulfur was performed at this stage. A final step of
purification on Fluorisil was then performed (40 ml
hexane+20 ml hexane:CH2Cl2, 95:5). The eluate was
evaporated, 10 Al of internal standard (PCB#155) was
added to 70 Al of the sample and 2 Al of this solution
was injected in the GC/MS.
Samples were analyzed using a Finnigan GC/MS
GCQ equipped with an AS9000 autosampler and a
CP-Sil 8 capillary column (50 m length, 0.25 mm id,
0.25 Am film thickness). Initial temperature of the
column was 90 jC. Temperature program was: (i)
increase to 180 jC at 15 jC min-1 and hold for 6 min,
(ii) increase to 220 jC at 4 jC min-1 and hold for 2
min and (iii) increase to 275 jC at 5 jC min-1. The
carrier gas was helium at a flow rate of 30 cm s-1.
Injection mode was splitless. Mass spectra were
acquired in electron impact in mode multiple reac-
tion monitoring.
Concentration of congeners #28, 52, 101, 118, 153,
138 et 180 were measured in each sediment sample
and in the different body compartments of the aste-
roids. Accuracy of the method was tested using
certified material reference from the Community Bu-
reau of Reference (sediments from the harbour Nova
Scotian in East of Canada and mackerel oil; CRM
no. 350). Analyzed CRM were always within 15% of
the mean certified values (except for congener 101
analyses that fell within 30% of the mean certified
value). The detection limits were between 0.01 and
0.1 ng g-1 dry wt., depending on the PCB congener.
2.3. Metal analyses
The concentrations of Zn, Cu, Cd and Pb were
measured in each of the five selected grain-size
fractions and in each asteroid body compartment,
according to the method described by Warnau et al.
(1995). Briefly, samples (0.4 g for sediments, 1 g for
asteroid body wall, and 0.5 g for other asteroid body
compartments) were digested with 65% nitric acid (2
ml g-1 dry wt. asteroid sample; Merck, p.a. grade).
Acid digestions were carried out at 20, 40, 60 and 80
jC during respectively 24, 6, 6 and 12 h, allowing
complete digestion of samples and avoiding any loss
due to overheating. Digests were diluted 12 times in
milli-Q water (Millipore) and filtered on glass micro-
fiber filters (Whatman GF/A). Metal concentrations
 B. Danis et al. / Science of the Total Environment 333 (2004) 149165 153

were determined by atomic emission spectrometry
using a Jobin-Yvon 38+ICP-AES. Accuracy of the
method was tested using certified reference material
(Mytilus edulis tissues, CRM no. 278, BCR). Ana-
lyzed CRM were always within 12% of the mean
certified values. Detection limits for Zn, Cu, Cd, and
Pb were, respectively, 0.002, 0.002, 0.001, 0.014 Ag
of metal per ml of digested sample.
2.4. Data analyses
For PCB analysis, it was assumed that the loss was
the same for the surrogate (PCB#103) as for the other
analyzed congeners, and the PCB concentrations were
corrected accordingly. PCB#28 is not reported be-
cause of unexplained interference with other com-
pounds. Everaarts et al. (1999) have encountered the
same type of analytical problems.
For metal analysis, emission intensities were con-
verted to concentrations (Ag g-1). Statistical differ-
ences between metal concentrations measured in
different grain-size fractions of sediments, asteroid
body compartments, and sampling stations were
assessed using one- or two-way analysis of variance
(ANOVA) followed by a multiple comparison test of
means (Tukey test) (Zar, 1996). The variability
explained by each factor considered was derived from
the sum of squares. Correlations between concentra-
tions of different metals measured in sediments or
asteroids were tested using simple linear correlation
procedures (Zar, 1996). Concordance of ranking of
the stations according to the contamination level
determined through sediment and asteroid analyses
were tested using Kendalls coefficient of concor-
dance (Zar, 1996). The level of significance for
statistical analyses was always set at a=0.05.
3. Results
3.1. PCBs in sediments
PCB concentrations were measured in the bulk
sediment. Data were analyzed using one-factor
ANOVA and are presented in Table 2. PCB concen-
trations varied significantly between the stations for
all the congeners except #180. The sum of six PCB
concentrations varied between 0.23 and 21.1 ng g-1
dry weight in sediment for the 19 stations. The lowest
concentrations were found in stations 330 and 140.

Table 2
PCB concentrations (meanFS.D.; Ag g-1 dry wt., n=6) in the bulk fraction of the sediments
Station A6PCB #52 #101 #118 #138 #153 #180
bc ab b b b b
120 4.79 F0.56 0.20 F0.01 0.53 F0.13 0.85 F0.27 0.94 F0.12 1.09 F0.17 0.66aF0.11
130 10.6abcF1.01 0.44aF0.05 1.16bF0.09 2.84aF1.36 2.03abF0.31 2.12abF0.40 1.60aF0.71
140 0.83cF1.03 0.07bF0.05 0.14bF0.10 0.28bF0.35 0.15bF0.19 0.21bF0.18 0.14aF0.14
150 1.06bcF0.62 nm nm 0.34bF0.16 0.25bF0.13 0.18bF0.13 0.19aF0.06
230 0.86bcF0.35 0.05bF0.01 0.11bF0.05 0.15bF0.10 0.21bF0.07 0.23bF0.08 0.07aF0.01
250 0.90bcF0.62 0.05bF0.02 0.08bF0.06 0.25bF0.16 0.18bF0.15 0.23bF0.12 0.16aF0.18
330 0.23cF0.09 0.05bF0.02 0.03bF0.01 nm 0.08bF0.03 0.06bF0.01 nm
700 1.93bcF0.74 0.23abF0.18 0.36bF0.25 0.27bF0.19 0.31bF0.09 0.45bF0.22 0.26aF0.22
710 1.90bcF1.48 0.15bF0.11 0.24bF0.16 0.51bF0.63 0.40bF0.19 nm 0.25aF0.08
Breskens 7.70abcF4.84 0.16bF0.08 1.48abF0.47 1.35abF1.34 2.29abF1.65 1.72abF0.91 0.63aF0.66
Knokke 2.93bcF3.64 0.05bF0.01 0.31bF0.18 0.53bF0.01 1.10bF1.48 0.80bF1.05 nm
Nieuwpoort 4.13bcF1.38 0.08bF0.03 1.15bF0.38 0.79bF0.32 0.94bF0.45 1.09bF0.37 0.14aF0.05
Oostende 1.05bcF0.19 0.07bF0.05 0.31bF0.13 0.31bF0.33 0.24bF0.09 0.34bF0.15 nm
Perkpolder 14.4abF13.8 0.16bF0.08 1.75abF1.41 1.22abF1.35 4.59abF4.59 3.90abF4.13 2.92aF3.72
S01 7.46bcF1.52 0.41aF0.23 0.92bF0.16 1.35abF0.32 1.36bF0.49 1.80abF0.39 1.07aF0.12
Scharendijke 3.94bcF0.29 0.24abF0.11 0.79bF0.23 0.73bF0.13 0.71bF0.10 0.88bF0.02 0.59aF0.30
Terneuzen 21.1aF11.3 0.16bF0.06 3.09aF1.53 1.83abF0.51 7.17aF5.04 5.80aF3.33 3.06aF2.72
Wenduine 2.98bcF2.36 0.03bF0.01 0.86bF0.25 0.67bF0.24 0.99bF0.90 0.92bF0.56 nm
ZG03 1.36bcF0.99 0.04bF0.03 0.12bF0.09 0.31bF0.34 0.32bF0.22 0.33bF0.21 0.15aF0.12
Superscripts indicate ranking of stations according to decreasing concentrations of a given PCB congener (a>b>c>. . .). Stations with
concentrations sharing a common superscript are not significantly different from each other (multiple comparison test of Tukey; a=0.05).
nm=Not measured.
154 B. Danis et al. / Science of the Total Environment 333 (2004) 149165
Stations close to the estuary and in front of Oostende
(Terneuzen, Perkpolder, 130) were the most contam-
inated. PCB patterns appeared identical for all the
stations: PCBs #153 and #138 were the most abun-
dant, whereas PCBs #52 and #180 were detected in
lower concentrations.
Analyzing data in stations located along transects
(Fig. 1) helped in obtaining more discriminative
information. One-way ANOVAs resulted in signifi-
cant differences in PCB concentrations between sta-
tions for each investigated congener. In Transect I,
Terneuzen exhibited significantly higher concentra-
tions of PCB#101 and of the sum of PCBs. Transect II
showed 130 and S01 were significantly more contam-
inated than other stations (all congeners except
PCB#52). No significant differences were found in
Transect III. In Transect IV, station 120 was signifi-
cantly more contaminated in most case. Station 130
was always significantly more contaminated than the
other stations in Transect V. Transect VI showed that
Knokke was significantly more contaminated
(PCB#180). Finally, in Transect VII, it appeared that
S01 was the most contaminated station (PCB#52) and
that Terneuzen was significantly more contaminated
in all the other cases.
3.2. PCBs in asteroids
Asteroids were found in 10 out of the 19 investi-
gated stations. Concentrations of six PCB congeners
(PCB#28 is not reported because of interference with
other compounds) were determined in four body
compartments (oral and aboral body walls, pyloric
caeca and gonads) (Table 3).
With regard to the sea stars, the sum of PCB
concentrations were found to be the highest in the
pyloric caeca, varying between 396 and 1695 ng g-1
total lipids. Oral and aboral body walls were generally
more contaminated than gonads. Pyloric caeca were
the most discriminative body compartment. Sea stars
collected in stations Knokke, Breskens, and Nieuw-
poort revealed the highest concentration levels. Indi-
viduals were less contaminated in stations Wenduine
and 230.
The PCB contamination profile was similar in
sediments and sea stars: PCBs #153 and #138
exhibited the highest concentrations, while congeners
#52 and #180 were the less concentrated.
PCB concentrations in the body compartments and
their lipid contents were compared and found to be
highly correlated (rz0.99, pV0.01), confirming that
PCB concentration in sea stars mainly depends on its
lipid content.
Possible correlations were also assessed between
PCB levels in sediments and asteroids, but no signif-
icant correlation could be found. A striking difference
between PCB concentration in sediments and aste-
roids is exemplified by stations Knokke and 330,
which both exhibit high PCB concentrations in aste-
roids while being among the less impacted station
when considering sediments alone.
The two latter stations show a very different
granulometry from the one of the other stations
(Table 4); fraction 250 500 Am is predominant in
Knokke and 330, whereas fraction 125 250 Am is
generally the most represented fraction in the other
stations. A good correlation (r =0.85, p=0.002) was
also found between the percentage of fraction 250
500Am and the concentration ratio between asteroids
and sediments (PCB concentration in asteroids lipid
wt.-1/PCB concentration in sediments dry wt.-1)
(Fig. 2).
3.3. Metals in sediments
The grain-size distributions of the different sedi-
ments are presented in Table 4. The fraction size
125 250 Am was dominant in 14 stations, fraction
250 500 Am in three stations and fractions 63
125 and 500 1000 Am only in one station.
Metal concentrations were measured in the differ-
ent sediment fractions (Table 5). Since some of the
investigated metals have similar physicochemical
properties, correlations between their concentrations
were determined in each grain-size fraction (data not
shown). Highly significant correlations ( pV0.0001)
were found between all metals in all sediment
fractions, except between Cu vs. Pb and Cu vs. Cd
in the <63 Am fraction. The highest correlation
coefficient values (rz0.92) were observed in the
125 250 and 63 125 Am fractions while the lowest
values were found in the <63 and 500 1000 Am
fractions.
Two-way ANOVA analyses showed that metal
concentrations differed significantly between stations
and grain-size fractions (the fraction <63 Am dis-
 B. Danis et al. / Science of the Total Environment 333 (2004) 149165 155

Table 3
PCB concentrations (meanFS.D.; ng g-1 total lipids, n=5 pools of three) in the different body compartments of the asteroid Asterias rubens
Station A6PCB #52 #101 #118 #138 #153 #180
(A) Oral body wall
230 431aF101 19.3aF16.1 63.4aF24.7 46.7aF10.6 169aF40.5 118aF11.6 nm
250 590aF222 28.4aF12.8 79.9aF31.5 76.6aF39.9 228aF85.0 176aF61.2 19.0aF7.59
330 836aF836 23.7aF23.7 119aF118 100aF100 314aF314 234aF234 36.0aF36.0
710 901aF233 16.5aF4.08 106aF13.4 148aF35.8 351aF122 280aF56.6 nm
Breskens nm nm nm nm nm nm nm
Knokke 1484aF594 63.9aF41.4 207aF54.1 148aF61.4 612aF250 443aF235 38.2aF7.56
Nieuwpoort 1128aF275 32.2aF13.5 157aF37.7 116aF21.9 437aF157 354aF124 38.5aF11.6
Oostende 1463aF719 39.0aF13.5 216aF86.3 130aF72.6 552aF310 445aF248 62.4aF39.2
Scharendijke 1202aF333 27.9aF11.8 150aF8.1 124aF32.3 484aF233 389aF164 51.2aF9.42
Wenduine 1045aF331 78.9aF45.3 178aF100 119aF24.4 377aF51.9 298aF110 29.9aF3.29

(B) Aboral body wall

230 778aF312 39.7aF28.9 130aF98.4 62.7aF11.8 298aF107 219aF75.5 27.7aF9.78
250 1015aF55 45.2aF16.9 105aF52.0 116aF75.4 410aF208 296aF160 55.0aF51.1
330 nm nm nm nm nm nm nm
710 nm nm nm nm nm nm nm
Breskens nm nm nm nm nm nm nm
Knokke 1046aF231 31.9aF17.2 154aF62.6 76.7aF39.3 475aF124 299aF52.3 28.6aF1.26
Nieuwpoort 919aF272 53.4aF16.3 70.7aF49.4 80.9aF23.6 393aF137 287aF94.4 62.7aF49.2
Oostende 843aF126 30.8aF9.97 124aF2.82 74.2aF19.1 335aF109 231aF29.4 46.6aF38.5
Scharendijke 1125aF132 62.5aF6.31 54.2aF6.25 100aF10.0 490aF119 361aF58.0 nm
Wenduine nm nm nm nm nm nm nm

(C) Pyloric caeca

230 562bF102 31.3bF0.47 102bF21.9 54.1aF32.8 189bF7.17 131bF51.8 27.5aF20.5
250 735bF233 29.0bF4.76 107bF32.3 110aF50.2 262bF103 197bF69.3 15.6aF9.43
330 592bF27.5 22.2bF9.79 61.0bF5.51 59.6aF13.9 223bF20.2 182bF19.4 24.7aF4.54
710 695bF440 42.5abF20.7 136bF62.5 87.4aF54.5 227bF135 191bF110 17.0aF11.4
Breskens nm nm nm nm nm nm nm
Knokke 1695aF658 88.6aF29.5 317aF119 150aF44.9 614aF270 463aF215 35.9aF20.8
Nieuwpoort 634bF261 22.8bF5.79 104bF47.4 78.3aF32.9 203bF101 162bF64.5 54.1aF32.7
Oostende 689bF338 24.9bF16.4 108bF65.9 74.9aF32.3 230abF140 205bF105 66.3aF71.9
Scharendijke 501bF246 24.9bF8.52 90.8bF44.3 53.9aF33.9 169bF106 120bF52.3 26.3aF15.4
Wenduine 396bF20.0 45.7abF11.6 62.5bF9.58 38.3aF20.2 134bF42.6 104bF11.4 nm

(D) Gonads
230 655bF164 56.9aF19.3 121bF42.6 71.9aF4.94 200bF70.6 169bF47.3 23.6bF4.37
250 561bF88.3 23.5aF8.48 83.3bF14.6 63.7aF32.1 213bF50.5 158bF27.4 16.1bF5.12
330 nm nm nm nm nm nm nm
710 356bF117 18.1aF5.45 50.5bF13.5 37.8aF8.74 145bF75.2 94.8bF39.4 nm
Breskens nm nm nm nm nm nm nm
Knokke 1473aF307 72.7aF65.9 294aF99.0 149aF44.4 546aF105 359aF36.2 50.9abF15.9
Nieuwpoort 727bF169 32.5aF14.5 129bF59.5 71.1aF28.5 237bF100 176bF71.6 75.7aF35.1
Oostende 688bF257 27.1aF15.0 117bF46.2 80.6aF46.1 219abF99.4 192bF88.2 nm
Scharendijke 653bF218 28.7aF14.7 136bF61.7 46.6aF21.1 251bF83.8 161bF73.6 nm
Wenduine 1325abF166 59.5aF4.76 229abF37.3 132aF22.1 519aF53.4 358abF72.9 37.1abF0.23
Superscripts indicate ranking of stations according to decreasing concentrations of a given congener (a>b>c>. . .). Stations with concentrations
sharing a common superscript are not significantly different from each other (multiple comparison test of Tukey; a=0.05). nm=Not measured.
156 B. Danis et al. / Science of the Total Environment 333 (2004) 149165

Table 4
Grain-size distribution (mean %FS.D., n=6) in the dried sediments for the 19 sampling stations
Station Grain-size fraction
500 1000 Am 250 500 Am 125 250 Am 63 125 Am <63 Am
120 1.48F0.45 10.5F0.82 72.9F4.43 6.30F1.53 8.84F2.57
130 0.02F0.01 0.89F1.19 28.7F2.55 46.3F3.69 24.1F4.31
140 0.49F0.22 7.54F6.73 80.4F9.85 5.46F2.35 6.13F3.35
150 1.25F0.51 46.7F6.57 49.3F6.59 1.91F0.42 0.85F0.30
230 1.38F0.55 19.2F3.45 78.5F3.42 0.78F0.15 0.18F0.05
250 72.2F8.89 20.1F6.04 5.43F3.08 1.99F1.42 0.33F0.21
330 11.7F4.37 72.4F4.37 15.4F5.20 0.42F0.19 0.10F0.04
700 0.14F0.07 0.67F0.40 39.1F11.6 27.5F3.36 32.6F9.66
710 13.8F29.2 14.8F11.4 53.8F35.5 27.8F48.2 25.7F49.5
Breskens 0.00F0.00 0.69F1.51 62.1F8.79 27.8F6.84 9.42F5.95
Knokke 11.2F1.95 69.1F2.46 19.4F1.94 0.25F0.02 0.04F0.04
Nieuwpoort 0.04F0.03 1.97F0.22 96.2F0.33 1.70F0.19 0.08F0.01
Oostende 1.62F0.55 23.1F2.92 74.7F3.26 0.53F0.04 0.06F0.01
Perkpolder 0.74F1.69 1.24F1.65 64.7F25.5 27.6F18.8 5.68F3.69
S01 1.38F0.89 58.4F8.08 25.4F4.75 7.72F2.31 7.17F2.08
Scharendijke 5.20F1.20 13.9F2.36 68.7F3.91 10.9F1.01 1.21F0.22
Terneuzen 0.01F0.00 4.59F2.00 89.0F5.12 5.34F5.62 1.04F0.22
Wenduine 3.79F1.00 50.4F5.65 45.5F6.54 0.20F0.03 0.06F0.07
ZG03 0.34F0.06 10.2F2.71 85.9F2.39 1.71F0.31 1.83F1.14
Predominant grain-size fractions are indicated in bold.

played the highest concentrations, except for Pb for
which fractions <63 and 500 1000 Am were the
most contaminated) (data not shown). The two
factors considered (sampling station and grain size)
and their interaction explained the largest part (66
83%) of the variability in metal concentrations
measured in sediments. However, 21 32% of that
variability were due to the interaction between the
two factors. Consequently, data were reanalyzed for
each sediment grain size, using one-way ANOVA,

Fig. 2. Correlation between concentration ratio (CR) of PCBs in the A. rubens pyloric caeca [CR=A6 PCBs in asteroids (ng g-1 lipids)/A6 PCBs
in sediments (ng g-1 dry wt.)] and the percentage of the grain-size fraction 250 500 Am in the sediments of the sampling stations (%).
 B. Danis et al. / Science of the Total Environment 333 (2004) 149165 157

Table 5
Metal concentrations (meanFS.D.; Ag g-1 dry wt., n=6) in the different grain-size fractions of the sediments
Station Zn Pb Cd Cu
(A) Fraction<63 lm
120 103F5.60ab 56.7F4.20ab 2.10F0.13a 13.3F1.00abc
130 86.7F11.6bc 47.6F7.20bc 2.00F0.26a 10.2F1.70bcd
140 68.6F6.60cde 36.2F2.80cd 1.40F0.16cd 7.90F1.40cd
150 59.1F7.60def 25.9F3.20def 1.00F0.13de 6.40F1.50cd
230 46.5F14.6efg 24.5F7.60def 0.90F0.28efgh 5.00F2.00d
250 48.6F8.60efg 22.4F3.70ef 0.70F0.14fgh 4.60F1.20d
330 36.8F20.4defg 22.4F13.9de 0.60F0.30efgh 3.70F2.30cd
700 45.7F3.96e 31.4F2.10de 1.79F0.15ab 6.70F0.66cd
710 52.9F13.4defg 29.1F7.10de 1.00F0.25ef 5.90F2.00cd
Breskens 72.6F16.7cd 28.0F7.00def 1.11F0.28de 7.60F2.70cd
Knokke 20.3F14.49fg 18.6F10.41def 0.40F0.24gh 4.20F5.79cd
Nieuwpoort 54.3F26.4defg 24.9F11.4def 0.80F0.33efgh 3.90F1.90d
Oostende 24.7F19.3efg 18.0F11.2def 0.40F0.26fgh 2.10F1.80d
Perkpolder 111F21.8a 68.0F14.0a 1.52F0.19bc 12.0F2.10bc
S01 120F7.50a 57.5F3.40a 1.90F0.13a 15.3F1.40ab
Scharendijke 83.9F13.6bc 34.0F7.00de 0.90F0.10efg 21.3F10.2a
Terneuzen 37.7F10.1fg 16.0F1.00fg 0.57F0.03gh 3.10F0.40d
Wenduine 21.5F12.7fg 20.2F9.56def 0.37F0.19h 4.10F3.04cd
ZG03 40.0F4.20fg 4.60F0.50g 0.90F0.07efgh 19.7F3.30a

(B) Fraction 63 125 lm

120 15.6F2.40b 11.8F1.50a 0.40F0.02a 1.20F0.01b
130 78.9F15.1b 45.6F8.70a 1.80F0.32a 9.50F2.40b
140 47.3F16.2c 28.5F7.00b 0.90F0.27bc 5.10F2.50cd
150 32.7F7.20cdefg 15.8F3.50cde 0.60F0.13de 3.40F1.50cdef
230 19.1F2.10efgh 11.4F1.40de 0.50F0.04def 1.50F0.40ef
250 16.8F2.10fgh 11.5F0.80de 0.30F0.02ef 1.30F0.30f
330 25.3F8.10defgh 14.6F2.40de 0.50F0.06def 2.20F0.50def
700 37.4F6.08cde 25.8F3.36bc 1.23F0.25b 5.80F1.28c
710 31.5F20.5cdefgh 18.1F7.10bcd 0.60F0.18def 3.20F2.00cdef
Breskens 34.3F10.1cdef 17.0F4.00cde 0.55F0.10def 3.30F1.30cdef
Knokke 14.3F1.66gh 16.4F4.55cde 0.41F0.01ef 1.50F0.62ef
Nieuwpoort 15.6F1.60fgh 11.0F0.40de 0.40F0.02ef 1.00F0.01f
Oostende 15.6F2.40fgh 11.8F1.50de 0.40F0.02ef 1.20F0.01f
Perkpolder 33.8F4.80cdef 19.0F4.00bcd 0.48F0.07def 3.60F0.80cdef
S01 103F6.00a 50.6F3.50a 1.60F0.09a 13.4F1.80a
Scharendijke 26.2F4.90defgh 13.0F8.00de 0.30F0.02ef 4.50F1.70cde
Terneuzen 12.8F1.40h 7.00F1.00e 0.26F0.02f 1.00F0.10f
Wenduine 16.5F3.53fgh 17.3F5.40cde 0.43F0.04ef 2.10F1.92def
ZG03 42.1F13.3cd 28.9F6.20b 0.80F0.20cd 5.40F2.00c

(C) Fraction 125 250 lm

120 28.7F7.90c 23.4F4.50cd 0.60F0.15c 3.20F1.10d
130 106F7.90a 64.9F3.30a 2.50F0.17a 13.7F1.00a
140 15.6F4.00def 15.8F7.50def 0.30F0.07de 1.10F0.50ef
150 5.80F0.70f 3.80F0.80g 0.10F0.01e 0.40F0.10f
230 14.3F1.70ef 4.30F0.40g 0.30F0.01de 0.60F0.01f
250 15.0F0.90def 8.70F0.60efg 0.30F0.02de 0.90F0.10ef
330 8.30F0.80f 5.00F0.20g 0.30F0.01e 0.70F0.10f
700 34.0F5.05c 25.9F3.43c 1.03F0.15b 5.60F1.16bc
710 11.4F2.00f 7.50F1.50fg 0.20F0.04e 0.90F0.30f
Breskens 27.8F9.90cd 14.0F4.00ef 0.47F0.12cd 2.80F1.30de
(continued on next page)
158 B. Danis et al. / Science of the Total Environment 333 (2004) 149165

Table 5 (continued)
Station Zn Pb Cd Cu
(C) Fraction 125 250 lm
Knokke 5.80F1.49f 3.30F0.44g 0.13F0.01e 0.30F0.05f
Nieuwpoort 11.4F0.90f 7.40F0.70fg 0.30F0.02de 0.60F0.10f
Oostende 7.40F0.60f 4.00F0.20g 0.20F0.01e 0.70F0.40f
Perkpolder 35.0F5.70c 17.0F2.00de 0.46F0.07cd 3.80F0.90cd
S01 63.1F20.1b 43.1F14.3b 1.20F0.28b 7.40F3.10b
Scharendijke 26.6F4.00cde 7.00F1.00fg 0.20F0.03e 2.30F0.50def
Terneuzen 8.90F0.90f 5.00F0.01g 0.23F0.02e 0.60F0.10f
Wenduine 6.00F0.79f 4.80F1.11g 0.16F0.01e 0.40F0.03f
ZG03 12.8F2.00f 9.70F0.70efg 0.30F0.03de 1.10F0.10ef

(D) Fraction 250 500 lm

120 37.0F11.3cd 26.0F7.10de 0.80F0.21cde 4.70F1.70d
130 120F13.2ab 66.6F4.90a 2.10F0.30a 16.3F2.30b
140 26.7F12.8cd 15.5F7.50efg 0.70F0.47cdef 2.40F2.00d
150 9.0F1.70cd 6.70F3.30gh 0.20F0.03gh 0.70F0.20d
230 13.0F2.60cd 4.20F1.40h 0.30F0.05gh 0.90F0.20d
250 11.8F1.40cd 6.90F0.70gh 0.10F0.03h 0.70F0.10d
330 7.80F1.10cd 7.40F5.30h 0.30F0.02gh 0.70F0.30d
700 36.1F8.65cd 34.5F5.61cd 1.10F0.25c 6.80F0.89cd
710 18.1F5.30cd 10.4F2.90gh 0.30F0.08fgh 1.30F0.80d
Breskens 95.1F45.0b 45.0F15.0bc 1.12F0.41c 13.2F7.40bc
Knokke 3.50F0.85d 3.30F0.24h 0.13F0.01h 0.30F0.02d
Nieuwpoort 19.4F1.50cd 16.1F1.80efg 0.50F0.04defg 1.50F0.20d
Oostende 6.30F0.70d 5.30.0F1.60gh 0.20F0.01gh 0.40F0.01d
Perkpolder 149F51.1a 50.0F10.0b 1.62F0.26b 27.2F14.9a
S01 34.9F5.00cd 22.8F4.50def 0.90F0.07cd 2.90F0.70d
Scharendijke 39.4F15.5c 15.0F6.00efg 0.34F0.16efgh 4.50F2.20cd
Terneuzen 20.3F9.40cd 11.0F6.00fg 0.47F0.16defgh 2.20F1.30d
Wenduine 4.70F0.85d 4.50F0.41gh 0.18F0.02gh 0.30F0.03d
ZG03 31.2F6.30cd 22.9F3.80de 0.80F0.15cdef 3.60F1.00d

(E) Fraction 500 1000 lm

120 40.1F10.9bcd 42.3F4.30bc 1.20F0.22a 6.20F1.60bcd
130 99.0F17.2ab 85.0F34.5ab 1.20F0.32a 13.4F3.00b
140 53.3F49.9bcd 28.7F24.5bc 0.88F0.81ab 7.10F6.10defgh
150 65.7F17.5abc 34.6F7.90bc 1.30F0.32a 9.30F2.90bc
230 28.2F3.60cd 15.8F3.20c 0.60F0.09ab 2.80F0.80efgh
250 31.9F5.50bcd 20.7F1.60c 0.62F0.06ab 3.10F0.75defgh
330 25.6F5.10cd 24.5F2.60bc 1.30F0.24a 1.80F0.30fgh
700 29.8F14.3bcd 37.0F8.55bc 0.88F0.16ab 5.70F1.84cdef
710 37.8F8.10bcd 32.3F3.20bc 1.20F0.12a 5.40F1.40cdefg
Breskens 53.3F26.8d 105F104a 0.75F0.41b 7.20F2.40cde
Knokke 9.80F0.82cd 15.2F0.78c 0.58F0.04ab 1.00F0.07gh
Nieuwpoort 25.8F17.2cd 23.7F4.20bc 0.87F0.17ab 1.80F0.50fgh
Oostende 16.0F1.30cd 26.2F2.60bc 0.98F0.09a 1.90F0.27fgh
Perkpolder 104F29.7a 38.0F7.00bc 1.12F0.21a 18.6F5.50a
S01 65.3F33.4abc 45.2F24.4bc 1.40F0.32a 7.50F3.10cd
Scharendijke 34.8F15.7 24.0F16.0 0.27F0.10 21.3F45.7
Terneuzen 2.10F19.6cd 28.0F8.00c 0.59F0.07ab 1.80F0.90h
Wenduine 14.3F1.39cd 23.1F1.01bc 1.04F0.08a 1.30F0.05gh
ZG03 28.2F9.50cd 31.7F4.4bc 1.30F0.16a 3.80F1.40defgh
Superscripts indicate ranking of stations according to decreasing concentrations of a given metal (a>b>c>. . .). Stations with concentrations
sharing a common superscript are not significantly different from each other (multiple comparison test of Tukey; a=0.05).
 B. Danis et al. / Science of the Total Environment 333 (2004) 149165
followed by the multiple comparison test of Tukey.
Results of the one-way ANOVAs showed significant
differences in metal concentrations among stations
for each investigated element and each sediment
grain-size fraction (Table 5). Although the contam-
ination level of several stations was not well con-
trasted, some stations (viz. Perkpolder, SO1 and
130) generally exhibited significantly higher metal
concentrations than the other ones, whereas other
stations (Knokke, Terneuzen, Wenduine, Oostende
and Nieuwpoort) were generally ranked among the
less contaminated ones.
When data were analyzed for stations located
along transects parallel or perpendicular to the coast
(see Fig. 1), a better discrimination between stations
was obtained. In Transect I, Perkpolder and Breskens
exhibited significantly higher concentrations of Cu
(grain size 500 1000 Am), of Cd (grain size 250
500 Am) and of Zn (grain size 250 500 Am).
Transect II showed 130 and S01 were significantly
more contaminated than other stations (Cu: 250
500, 125 250 and 63 125 Am; Cd: 250 500 and
125 250 Am; Pb: all except the finest fraction; Zn:
all except 500 1000 Am). ZG03 was the most
contaminated station of Transect III, showing the
highest concentrations in Cu (250 500 and<63
Am), Cd (250 500 Am), Pb (250 500 and 63 125
Am), Zn (250 500 Am). In Transect IV, station 120
was significantly more contaminated by Cu (500
1000, 125 250 and 63 125 Am), Cd (in the three
finest fractions), Pb (all except 250 500 Am) and Zn
(the three finest fractions). Station 130 was always
significantly more contaminated than the other sta-
tions (except for Cd, 500 1000 Am). Transect VI
showed that station 150 was significantly the most
contaminated (in every case, except Cu: 250 500
Am; Cd: 125 250 Am; Pb: 250 500 and 63 125
Am; Zn: 125 250 Am). In Transect VII, it appeared
that SO1 was the most contaminated station in
almost every case (except Cu: 500 1000 and 250
500 Am; Cd: 500 1000 and 250 500 Am; Pb: 500
1000 and 250 500 Am; Zn: 500 1000 Am).
3.4. Metals in asteroids
Despite a constant sampling effort, asteroids were
found in only 10 out of the 19 stations investigated,
viz. stations 230, 250, 330, 710, Breskens, Knokke,
159
Nieuwpoort, Oostende, Wenduine and Scharendijke
(see Table 1 and Fig. 1).
Concentrations in Zn, Pb, Cd, and Cu were mea-
sured in five body compartments (oral and aboral body
wall, pyloric caeca, digestive tract and gonads) (Table
6). Possible correlations between concentrations of
different metals in a given body compartment were
determined (data not shown). In contrast with what
was observed in sediments, only few significant cor-
relations were found in asteroids. In addition, signif-
icant correlation coefficients were generally much
lower than those calculated for metals in sediments.
Two-way ANOVA analyses showed that metal
concentrations differed significantly among stations
and among body compartments for all investigated
elements. In particular, the multiple comparison test
of Tukey (data not shown) indicated that Cd and
Cu were always more concentrated in pyloric
caeca, while Zn tended to be preferentially concen-
trated in the aboral body wall; no clear trends was
found for Pb. Two-way ANOVA also showed that
the two factors considered (station and body com-
partment) and their interaction were responsible for
41 86% of the variability in metal concentrations
measured in asteroids. Except for Pb, the largest
part of variation was always associated to the
body compartment factor. Interaction between
station and body compartment factors was
always significant and accounted for an important
part of the observed variability (18 35%), indicat-
ing that differences in station contamination varied
according to the considered body compartment.
Therefore, data had to be reanalyzed using one-
way ANOVA, considering separately each body
compartment. The latter analyses showed that, ex-
cept for Cd, Pb and Zn in the digestive tract and
for Pb and Zn in the gonads, significant differences
in metal concentrations occurred among stations
( pANOVA always <0.03). Among the asteroid body
compartments, the oral body wall appeared to be
the most discriminative. The ranking of the stations
according to decreasing metal concentrations dif-
fered according to the metal and the body com-
partment considered (Table 6).
However, asteroids from six sampling stations
(Breskens, Knokke, Wenduine, Scharendiijke, Oos-
tende and 330) tended generally to exhibit the highest
metal concentrations.
160 B. Danis et al. / Science of the Total Environment 333 (2004) 149165

Table 6
Metal concentrations (meanFS.D.; Ag g-1 dry wt., n=5 pools of three) in the different body compartments of the sea star Asterias rubens
Station Zn Pb Cd Cu
(A) Oral body wall
230 38.1F25.8c 0.85F0.63bc 0.10F0.06cde 0.38F0.22abc
250 86.9F19.9ab 1.47F0.44ab 0.16F0.04cd 0.51F0.19abc
330 59.1F8.7bc 2.11F0.97a 0.21F0.02bc 0.91F0.14a
710 73.3F13.1abc 1.62F0.07ab 0.17F0.03bcd 0.42F0.06bcd
Breskens 98.5F13.5a 1.77F0.43ab 0.31F0.06a 0.55F0.30abc
Knokke 73.1F5.9abc 2.00F0.34ab 0.25F0.02ab 0.75F0.14ab
Nieuwpoort 44.4F23.3c 0.16F0.02c 0.03F0.02e 0.02F0.00e
Oostende 68.9F16.1abc 1.77F0.18ab 0.19F0.01bc 0.25F0.24de
Scharendijke 54.6F19.2bc 0.46F0.58c 0.09F0.08de 0.07F0.11e
Wenduine 82.2F13.0ab 1.37F0.13ab 0.17F0.02cd 0.12F0.09de

(B) Aboral body wall

230 109F65.8d 0.36F0.19bc 0.22F0.11bc 0.54F0.67c
250 156F34.7ab 0.26F0.03ab 0.27F0.06bc 0.04F0.00c
330 91F24.6e 1.15F0.16c 0.31F0.10abc 0.04F0.00c
710 189F35.5ab 0.52F0.12bc 0.28F0.06bc 1.45F0.62a
Breskens 161F28.7ab 0.91F0.13abc 0.47F0.10ab 0.02F0.00c
Knokke 135F37.8b 1.16F0.53ab 0.51F0.19a 0.13F0.17c
Nieuwpoort 127F26.5b 0.52F0.28bc 0.25F0.08c 0.03F0.01c
Oostende 120F26.2b 0.92F0.72abc 0.26F0.03c 0.03F0.01c
Scharendijke 210F50.0a 0.73F0.20abc 0.25F0.06c 0.03F0.01c
Wenduine 108F21.1c 1.43F0.32a 0.27F0.05bc 0.26F0.19b

(C) Pyloric caeca

230 53.0F64.5b 0.95F0.73abc 1.31F1.15ab 5.12F4.15c
250 45.1F8.80b 1.27F0.25abc 0.83F0.52b 5.45F0.64c
330 58.3F9.30b 1.19F0.23bc 0.60F0.23b 19.0F9.70bc
710 149F12.4ab 0.63F0.16c 0.92F0.25ab 7.63F3.37c
Breskens 89.4F10.6b 2.03F0.32a 1.06F0.64ab 30.3F17.6b
Knokke 73.1F4.90b 1.51F0.13ab 2.36F1.40a 16.4F2.97bc
Nieuwpoort 57.2F12.0b 1.16F0.70bc 1.11F0.38ab 9.00F1.44c
Oostende 119F27.9b 1.18F0.10bc 1.62F0.78ab 11.6F2.18c
Scharendijke 282F186a 1.22F0.58bc 0.56F0.18b 51.1F14.4a
Wenduine 88F9.3b 1.47F0.16abc 0.62F0.13b 11.8F0.75bc

(D) Gut
230 37.6F37.5c 0.92F0.33c 0.19F0.08c 3.76F3.19ab
250 79.9F4.60c 1.09F0.75c 0.21F0.08c 4.92F1.06ab
330 70.2F8.60c 2.11F2.09c 0.24F0.13c 4.84F1.72ab
710 68.8F2.70c 1.15F0.66c 0.22F0.08c 4.25F1.67ab
Breskens 72.9F6.30c 1.36F0.24c 0.26F0.09c 4.92F2.18ab
Knokke 67.5F4.60c 0.92F0.09c 0.19F0.07c 2.33F0.28b
Nieuwpoort 59.3F1.80c 1.08F0.50c 0.19F0.04c 7.11F2.70a
Oostende 58.6F30.1c 1.20F0.46c 0.16F0.05c 2.72F1.22ab
Scharendijke 81.7F16.4c 1.86F2.00c 0.12F0.02c 6.52F3.48a
Wenduine 70.9F26.0c 1.68F0.76c 0.19F0.04c 3.52F1.65ab

(E) Gonads
230 37.6F15.9c 0.59F0.37c 0.09F0.07ab 0.74F0.73b
250 55.0F16.7c 1.02F0.03c 0.06F0.01b 1.25F0.21b
330 37.2F25.9c 1.44F1.36c 0.12F0.08ab 0.90F1.11b
710 23.0F25.3c 0.96F0.10c 0.05F0.01b 1.52F1.19b
 B. Danis et al. / Science of the Total Environment 333 (2004) 149165
Table 6 (continued)
Station Zn
(E) Gonads
Breskens 32.3F9.1c
Knokke 42.4F29.8c
Nieuwpoort 56.7F7.3c
Oostende 39.7F25.7c
Scharendijke 78.1F16.7c
Wenduine 40.4F17.7c
Superscripts indicate ranking of stations according to decreasing concentrations of a given metal (a>b>c>. . .). Stations with concentrations
sharing a common superscript are not significantly different from each other (multiple comparison test of Tukey; a=0.05).
3.5. Comparison between metal levels in sediments
and asteroids
Comparisons between data from sediment and
asteroid analyses were performed in considering only
stations in which both sediments and asteroids were
collected. Some significant correlations were found
between metal concentrations measured in sediments
and in asteroid body compartments (oral and aboral
body walls and pyloric caeca). However, correlation
coefficients were generally low (maximum value of
r =0.74).
Station rankings using sediment and asteroid data
were compared using Kendalls coefficient of concor-
dance. Results of these tests did not show any signif-
icant concordance. This lack of ranking concordance
was probably mainly due to several stations display-
ing intermediate and poorly contrasted contamination
levels.
4. Discussion
Determination of the grain-size distribution in
collected sediments showed that the fraction 125
250 Am (fine sand) was predominant in most of the
considered stations. Its degree of predominance varied
significantly from one station to another, suggesting
that local hydrodynamics are variable.
PCB analyses in bulk sediments showed that the
lowest concentration levels were found in the offshore
stations (e.g., stations 330 and 250) while the highest
levels were measured in stations under direct influ-
ence of the Scheldt river (Terneuzen, Perkpolder, 130,
S01). These results are in agreement with those found
in other studies (Laane et al., 1999) in the Dutch
161
Pb Cd Cu
0.58F0.06c 0.11F0.02ab 2.04F0.58b
1.48F0.48c 0.08F0.04b 2.07F0.96b
1.01F0.45c 0.18F0.04a 6.66F2.31a
0.76F0.20c 0.11F0.03ab 1.78F0.85b
1.75F1.87c 0.12F0.01ab 6.18F3.24a
0.88F0.13c 0.11F0.04ab 1.93F0.71b
coastal zone. Station 130 is localized where the
Scheldt gyre meets North Atlantic currents, giving
place to locally favorable conditions for the sedimen-
tation of heavily contaminated particles swept by the
Scheldt. Stations Perkpolder, Terneuzen and S01 are
situated in the Scheldt and consequently receive
highly contaminated flows. Considering PCB conge-
ners separately, #101 was the most discriminating one
(although not the most abundant). As found in previ-
ous studies (e.g., Stebbing et al., 1992), the most
abundant congeners in sediments were #153 and
#138.
The use of transects (parallel or perpendicular to
the coast, Fig. 1) brought a more discriminative
approach, as a smaller number of stations were
considered. This helped in enlightening more subtle
differences between contamination levels in the sam-
pling stations. Perpendicular transects (Transects IV,
V, and VI) were particularly useful in showing the
contamination gradient direction; intermediate sta-
tions were always more contaminated. Highest PCB
concentrations were measured along the halfway
transect (Transects II), which includes stations under
direct influence of the Scheldt (Lizen, 1990; Bayens,
1998). This shows that PCB contamination from the
Scheldt spreads out southerly, along the coast.
PCB concentrations measured in asteroids were
comparable to those measured in previous studies
(Everaarts et al., 1998; den Besten et al., 2001). PCBs
were distributed selectively between body compart-
ments of the sea stars. Generally, pyloric caeca
exhibited the highest concentrations. No significant
difference was found between contaminant levels in
aboral and oral body walls, although only the oral
body wall is in direct contact with sediments. This
observation was already made in experimental expo-
162 B. Danis et al. / Science of the Total Environment 333 (2004) 149165
sure conditions, and is probably due to the fact that
contamination of sea stars occurs directly by contact
with sediments itself but also by resuspended particles
(Danis et al., 2003).
Sampling sites were ranked according to their PCB
concentration level in sediments and in asteroids. No
rank concordance was found, probably because sta-
tions such as Knokke, 330 or Wenduine presented
surprising differences between relative contamination
of asteroids and sediments. These three stations were
the only ones where fraction 250 500 Am was
dominant (together with station S01, where no sea
stars were found). The grain-size distribution varied
from one station to another and bioavailability of
contaminants is known to vary according to grain size
(Luoma and Carter, 1991). Therefore, the relative
importance of these fractions might influence the
exposure of asteroids to the contaminants. A signifi-
cant correlation (r=0.85) was found between asteroid
concentration ratios (CRs) and the percentage of the
250 500 Am grain-size fraction in sediments. This
suggests that contaminants associated to this grain-
size fraction are more readily available for asteroids.
Another cause for the lack of concordance between
ranking of stations obtained via sediment and asteroid
analysis could be that asteroid specimens were found
only in some sampling stations, and particularly that
none was found along Transect II, where sediments
were the most contaminated. The sediment type
(muddy) along this transect is not appropriate for
asteroid populations (Hayward and Ryland, 1990).
The fact that sea stars were found in stations where
sediment contamination levels were not very well
contrasted might also have affected statistical tests
(i.e., rank concordance). Other authors have encoun-
tered the same problem (e.g., Guns et al., 1999).
Metal analysis in the different grain-size fractions
of the sediments showed selective distribution. The
finest fraction (<63 Am) displayed the highest metal
concentrations. This observation was expected be-
cause of the high surface/volume ratio of these par-
ticles. The fraction 125 250 Am was always the least
concentrated; because it is the dominant fraction in
most of the stations, competition for adsorption sites
is probably lower. Surprisingly, fraction 500 1000
Am (coarse sand) was the most contaminated by Pb
and was highly contaminated by Cd. This observation
might be due to this fractions high content in mollusc
shell and echinoderm test debris (Poulicek, 1982).
Indeed, these metals are known to have a high affinity
for calcitic skeletons (Sturesson, 1976; Shen and
Boyle, 1988; Temara et al., 1998a).
Significant differences were found between the
sampling stations: stations such as S01 and 130 were
the most contaminated. S01 is located in the middle of
the Scheldt mouth, directly undergoing the effects of
riverine and tidal currents, though protected from
North Atlantic currents. A good explanation for its
high contamination by heavy metals is that this station
is the most saline one among those sampled in the
Scheldt. Indeed, it is a well-described phenomena that
organic matter tends to flocculate when salinity
increases and that metals have a tendency to complex
with suspended organic matter (NSTF, 1993a; Verlaan
et al., 1998). In addition, station 130 is located in a
low-energy area, where the Scheldt gyre contacts the
Northwards Atlantic currents: their interaction leads to
local conditions allowing sedimentation of very fine
particles swept by the Scheldt (Lizen, 1990). Coastal
stations, especially Knokke, Oostende and Wenduine
exhibit relatively low metal concentrations: they are
probably partly protected from the Scheldt particulate
matter contribution.
As in the case of PCBs, the use of transects for
metal analysis enlightened more subtle differences
among contamination levels in the sampling stations.
As for PCBs, the highest metal concentrations were
measured along Transect II.
The metal measurements in asteroids showed that
investigated elements are selectively distributed
among body compartments. Cu and Cd are more
concentrated in the pyloric caeca; Zn is more effi-
ciently concentrated in the aboral body wall, while Pb
does not seem to have a preferential target com-
partment. The latter observation was somehow unex-
pected. Indeed, data available in the literature
generally report that Pb tends to be more efficiently
incorporated in calcitic skeletons (Kersten and
Kroncke, 1991; Temara et al., 1997, 1998b).
Differences in metal contamination among stations
were found in each asteroid body compartment.
Among these compartments, three of them (pyloric
caeca, oral body wall, and aboral body wall) showed
better discrimination among sampling stations. Station
ranking was obtained from sediment and tissue anal-
yses. As in the case of PCBs, no concordance was
 B. Danis et al. / Science of the Total Environment 333 (2004) 149165
found between ranking obtained for sediments and
asteroids, indicating that using a physicochemical
approach (sediments) might not be sufficient to char-
acterize contamination as it does not take into account
bioavailability.
Significant correlations were found between metal
concentrations measured in each sediment grain size
and in each asteroid body compartment. The highest
correlations were found for grain-size fraction <63
Am. This indicates that sediments are indeed one
possible metal contamination source for asteroids,
but recent experimental data (Warnau and Danis,
unpublished) showed that the main sources of metal
contamination for A. rubens are seawater and food,
sediments contributing only a very small proportion of
global exposure. The body compartment exhibiting
the largest number of correlations was the aboral body
wall, although this compartment is not in direct
contact with sediments. Aboral body wall can there-
fore be contaminated in two ways: seawater and
translocation of contaminants from other organs or
tissues. These two sources coexist: some authors have
shown that A. rubens concentrates heavy metals from
seawater (den Besten et al., 1990; Temara et al., 1996,
1998a; Warnau et al., 1999), and translocation pro-
cesses are well documented in echinoderms (Temara
et al., 1996, 1998b; Warnau et al., 1995, 1996, 1999).
Using A. rubens in this study allowed us to take
into account the multiple contamination sources and
the variation of bioavailability of metals depending on
sediments. Investigating biota brings complementary
information that is necessary to tackle the ecological
consequences of contamination. A. rubens helped in
identifying a group of more contaminated stations,
different from that which would have resulted from a
study limited to sediment analysis. Analysis of biota
and sediments is however more demanding than the
latter approach alone because it implies finding organ-
isms. This constraint can be solved, e.g., by using
active monitoring or bioassays (den Besten et al.,
2001).
In sediments, concentrations of Zn, Pb, Cd and Cu
are in the range generally reported (e.g., Bayne et al.,
1988; Everaarts and Fischer, 1989; Stebbing et al.,
1992; NSTF, 1993a,b). However, Cd concentrations
along Transect II are in the upper part of the ranges
generally reported in sediments from the North Sea.
This transect can therefore be considered as relatively
163
contaminated, at least by Cd. Concentrations mea-
sured in the asteroids are also in the range found in
literature for the same species (Riley and Segar, 1970;
Bryan, 1984; Everaarts and Fischer, 1989; Temara et
al., 1997). Sampling stations where asteroids were
found are not particularly contaminated by heavy
metals, implying a certain weakening of statistical
tests (no concordance between sediment and asteroid
contamination levels).
5. Conclusion
Variations in contaminant levels in sediments from
the Southern Bight of the North Sea are mainly
explained by hydrodynamic factors: stations with
low residual currents are often the most contaminated
ones. Sediments are a source of contamination for
asteroids, although it may not be considered as the
main one. The use of A. rubens has shown that this
species helps in distinguishing the contamination state
of selected stations while taking into account contam-
inant bioavailability. Some body compartments appear
to be better bioindicators, i.e., pyloric caeca. Informa-
tion obtained by the use of asteroids does not compare
with that obtained by the use of sediments, and this
highlights the observable differences between an
abiotic indicator (sediments) and a biotic indicator
(asteroids), the latter integrating the bioavailability of
contaminants.
Acknowledgements
Research supported by a Belgian Federal Research
Program (SSTC, Contract MN/11/30). Grateful thanks
are due to Ch. De Ridder, C. De Amaral, G. Coteur
(ULB), P. Gosselin (UMH) and G. Radenac (Univ. La
Rochelle, France) for helpful assistance in offshore
and sea shore samplings. M. Warnau and Ph. Dubois
are, respectively, Honorary Research Associate and
Research Associate of the National Fund for Scientific
Research (NFSR, Belgium). The IAEA-Marine Envi-
ronment Laboratory operates under a bipartite agree-
ment between the International Atomic Energy
Agency and the Government of the Principality of
Monaco. Contribution of the Centre Interuniversi-
taire de Biologie Marine (CIBIM).
164 B. Danis et al. / Science of the Total Environment 333 (2004) 149165
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