Environmental To,ricology and Chemisrry, Vol. 7, pp. 587-592, 1988 0730-7268/88 $3.00 + .

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Printed in the USA. Permmnn Prew d c

Short Communication
RESPONSES OF POLYCHAETES TO
CADMIUM-CONTAMINATED SEDIMENT:
COMPARISON OF UPTAKE AND BEHAVIOR
B. L. O L L A , *V.
~ B. ESTELLE,$
R. C. S W A R T Z , ~
G. BRAUN,$and A. L. STUDHOLME~
TCooperative Institute for Marine Resources Studies, National Marine Fisheries Service,
Northwest and Alaska Fisheries Center, $Cooperative Institute for Marine Resources Studies,
5U.S. Environmental Protection Agency, Mark 0. Hatfield Marine Science Center,
Newport, OR 97365; and ?National Oceanic and Atmospheric Administration,
National Marine Fisheries Service, Northeast Fisheries Center, Sandy Hook Laboratory,
Highlands, NJ 07732

(Received 4 September 1987; Accepted 22 December 1987)

Abstract -The burrowing behavior of three polychaete species, Nereis virens Sars, Glycera dibran-
chiata Ehlers and Nephtys caeca Fabricius, was examined after variable exposure periods (7, 14,
21 and 28 d) to sediment contaminated with a cadmium concentration of about 40 mg kg-' (dry
weight). Most comparisons of burrowing times and rates between exposed and unexposed worms
were not statistically significant. Those that were significant were relatively minor, e.g., a maxi-
mum reduction in burrowing time of 6.3 s, and probably would have little ecological relevance. The
feeding behavior of G. dibranchiata on the polychaete Euzonus mucronata was not affected by a
28-d exposure to cadmium-contaminated sediment. Uptake of cadmium varied among the three spe-
cies. After 28 d of exposure, cadmium body burdens were highest for N. virens (319 mg kg-I),
intermediate for G. dibranchiata (120 mg kg-') and lowest for N. caeca (39 mg kg-I). This poten-
tial for bioaccumulation and survival in cadmium-contaminated sediment indicates that some poly-
chaete species like N. virens may be an amplified source of cadmium for their crustacean, fish and
avian predators.

Keywords- Polychaetes Cadmium Behavioral effects Uptake

INTRODUCTION (a) examine the effects of sublethal concentrations

Cadmium pollution of the marine environment
has the potential to affect organisms at all trophic
levels [l-31. Although most cadmium toxicity tests
have been conducted with pelagic species, chronic
effects are likely to be caused by cadmium that has
been incorporated into sediment [3]. Infaunal poly-
chaetes typically survive exposures to relatively
high concentrations of cadmium, but they accu-
mulate high levels in their tissues [2,4,5]. Sublethal
effects due to the bioaccumulation of cadmium
may reduce survival of polychaetes under natural
conditions [5]. Induced changes in behavior have
been shown to be a sensitive and ecologically rele-
vant measure of pollution [6,7]. For example, sub-
lethal effects of oil on the burrowing behavior of
Nereis virens and Glycera dibrunchiata increased
the vulnerability of these polychaetes to predators
[8,9l.
The objectives of the present research were to
whom correspondence may be addressed.
587
of cadmium-contaminated sediment on the bur-
rowing behavior of N. virens, G. dibranchiatu and
Nephtys cuecu Fabricius, and on the feeding be-
havior of G. dibranchiata, and (b) compare the
bioaccumulation of cadmium by these three spe-
cies under identical exposure conditions.
MATERIALS AND METHODS
Collection and maintenance of
experimental animals
Nereis virens (mean, 3.49 k 1.92 SD g wet weight;
127 k 36 SD mm total length) and GIyceru dibrun-
chiata (mean, 3.01 +- 1.38 g; 128 5 28 mm) were
collected in Spermaceti Cove, Sandy Hook Bay,
New Jersey, and shipped by air to Newport, Ore-
*
gon. Nephtys caeca (mean, 6.59 2.52 g; 174 29 *
mm) were collected from Yaquina Bay, Oregon.
Prior to exposure, the worms were held without
f o o d for 3 to 18 d in sand-filled trays submerged
in flow.through tanks, Both the experimental and
the holding facilities were maintained at 15C and
588 B. L.
20 to 25 ppt salinity and on a 12-h light:l%-h dark
photoperiod.
Collection and preparation of test sediment
Sediment was collected from Yaquina Bay using
a 5-liter dredge 6 d prior to an experiment. The
sediment was sieved (0.5 mm screen) into filtered
(0.45 pm) seawater (25 ppt). After sieving, the
sediment was allowed to settle overnight and was
stored at 4C. The test sediment was a well-sorted,
fine sand with a mean total volatile solids content
of 1.5% (loss of weight on ignition at 45OOC).
Cadmium stock solutions were prepared by dis-
solving reagent-grade cadmium chloride (CdCI2.
2; H20) into filtered seawater and added to sedi-
ment according to the methods of Swartz et al.
[3]. Three hundred milliliters of filtered seawater
was added to glass mixing bottles containing 4
liters (7.2 kg) of cadmium-contaminated sediment,
placed on a rolling mill and rolled at 15 rpm for
4 h. The contents of each bottle were then mixed,
and 2 liters (3.6 kg) of sediment was added to each
of 13 6-liter glass aquaria (24.5 x 17.5 cm) that
had been acid-washed. Two liters of filtered (0.45
pm) seawater (25 ppt) was added to each aquar-
ium, and after 12 to 15 h, three worms were intro-
duced into each aquarium. One liter of overlying
seawater was replaced in each aquarium daily.
Three series of 28-d experiments were per-
formed, each on one species of worm. Treatments
in all series of experiments consisted of aquaria
with cadmium-contaminated (37.3 to 47.1 mg kg-'
initial concentration) sediment and overlying sea-
water; controls consisted of aquaria with uncon-
taminated sediment and overlying seawater. One
aquarium in each treatment was sampled at 0 d,
and three aquaria from control and contaminated
treatments were sampled at 7, 14, 21 and 28 d.
Analytical chemistry
For each species of worm, one experiment was
performed in which four types of samples were
collected for cadmium analyses using the methods
of Swartz et al. [3]: (a) overlying seawater, (b)
interstitial water, (c) bulk sediment and (d) poly-
chaetes for body burden measurements. All cad-
mium concentrations in sediment and worms are
reported on a dry weight basis. One aquarium
from each cadmium treatment and control was
sacrificed at the beginning of an experiment to
measure initial cadmium concentrations in overly-
ing water, interstitial water, bulk sediment and
worms. Thereafter, the four types of samples were
taken at 7-d intervals from three control and three
OLLA ET AL.
contaminated aquaria and were analyzed follow-
ing the procedures of Swartz et al. [3].
For body burden analyses, worms were sieved
from sediment and placed in aquaria containing
filtered seawater for 24 h to provide sufficient time
for the digestive tract to clear. Worms were removed
from the seawater, dried in an oven overnight at
103"C, weighed to determine dry weight and then
digested with heated, concentrated HN03. The
resulting clear liquid was evaporated to less than
1 ml, and 5 ml of hot 1 N HN03 was added be-
fore the solution was filtered into a 25-ml flask
and diluted to 25 ml with distilled, deionized water.
Worm extracts were analyzed by direct-aspiration
atomic absorption spectrophotometry.
Burro wing behavior in
cadmium-contaminated sediment
Three worms were introduced into each aquar-
ium ( N = 36 each of exposed and unexposed
worms), and the time to bury and anchor the head
(Stage I) and the time to bury the remaining body
segments (Stage 11) were recorded after the meth-
ods of Olla et al. [8]. These stages are identified
merely to facilitate observations; they have no eco-
logical significance. Every 7 d for 28 d, nine ex-
posed and nine unexposed worms were removed
from their aquaria. Each worm was transferred
to an individual aquarium containing uncontami-
nated sediment, and Stage I and Stage I1 burrow-
ing times were recorded. Stage I1 burrowing times
were converted to burrowing rates (mm s-I) based
on the length of each worm. This conversion was
performed to normalize the data, since all worms
were not of equal length.
Statistical comparisons of burrowing times and
rates between contaminated and uncontaminated
sediment were made for the initial introduction
and reburial events using the Mann Whitney U test
1101.
Feeding behavior of cadmium-exposed
G. dibranchiata
Following 28 d in the aquaria, 20 worms from
cadmium-contaminated sediment and 20 worms
from uncontaminated sediment were removed and
placed in individual glass containers (22 X 12 X 7
cm) containing clean sediment. Three days later,
three live polychaetes, Euzonus mucronata, c d -
lected from the high intertidal regions of Lost
Creek Beach, Oregon, were placed into each con-
tainer with an individual G. dibranchiata. After
I d, the contents of each container were sieved
 Effects of cadmium on polychaete behavior 589

through a 1-mm screen and the number of E.
mucronata present recorded.
RESULTS
Cadmium sediment concentrations
In the controls, cadmium concentrations were
less than 0.2 mg kg-' dry weight, in control sedi-
ment, and below detection limits (less than 0.006
mg L-I) in interstitial and overlying water. Mea-
sured concentrations of cadmium in sediment and
in interstitial and overlying water varied among
treatments and over time (Table 1). Mass balance
calculations for the experiments showed that cad-
mium was taken out of the aquaria through the
daily replacement of half of the seawater, result-
ing in decreased concentrations of cadmium in
interstitial water and seawater in the aquaria. Cad-
mium concentrations in sediment ranged from 37.3
. t o 47.1 mg kg-' at the initiation of exposure and
decreased to 27.2 to 38.3 mg kg-' by the 28th
day. Cadmium concentrations in interstitial and
overlying water decreased during the experiments.
After 28 d, the mean cadmium concentration in
interstitial water was 21.1% of the initial mean
concentration.
Differences in cadmium concentrations among
experiments were greater for interstitial water than
for overlying water or sediment (Table 1). At the
initiation of the experiments, interstitial water cad-
mium concentrations were 14.5 to 14.7 mg L-'
for G. dibranchiata, 9.3 mg L-' for N.virens and
2.9 mg L-' for N. caeca.
Cadmium uptake and mortality
All three species accumulated cadmium (Fig.
1). After 28 d , uptake was highest in N. virens
(mean, 319 mg kg-') and appeared to be increas-
ing. In G. dibranchiata, uptake was markedly
lower (mean, 120 mg kg-' after 28 d), despite the
fact that interstitial cadmium level was higher than
that to which N. virens was exposed. In N. caeca,
uptake was lowest (mean, 39 mg kg-I) and ap-
peared to be reaching a constant level. Cadmium
body burdens in all three species held in uncon-
taminated sediment were less than 1.8 mg kg-'
and did not increase over time.
Mortality was negligible for all species in both
uncontaminated sediment (2070) and contaminated
sediment (4.5To).
Effects of cadmium on burrowing
Polychaetes placed in aquaria typically crawled
forward on the bottom, stopping occasionally to
probe the sediment with the prostomium. At the
initiation of burrowing, the prostomium was bur-

Table l . Cadmium concentration in sediment (mg kg-' dry weight) and in interstitial water and
overlying seawater (mg L-') during 28-d burrowing experiments with Nereis virens, Glycera dibranchiata
and Nephfys caeca, and during a 28-d feeding experiment with G. dibranchiata

Interstitial Overlying
Day Sediment water water

Burrowing experiment
N. virens 0 47.1 9.3 1.5
7 47.3 4.7 2.4
14 42.0 3.6 0.9
21 36.1 2.2 0.4
28 38.3 0.7 0.3
G. dibranchiata 0 38.4 14.5 0.9
7 31.8 13.1 2.4
14 28.5 4.6 1.3
21 30.2 3.0 0.7
28 27.2 1.1 0.7
N. caeca 0 37.3 2.9 1.1
7 37.0 2.0 1.6
14 36.1 1.5 0.4
21 36.8 1.o 0.1
28 37.0 1.7 0.0
Feeding experiment
G. dibranchiata 0 35.7 14,7 2.3
28 28.2 1.6 0.1
590 3. L. OLLAET AL.

Ir----T- A OExposed to
Cadmium
300 - 0 Not exposed
--
2 40
*
-F
I
260-
0 30
%
a
180-
w 20
V
z
0
0

n 10
0
al
v)
Y
0
W
r
I- 30
6
7 14 21
CADMtUM EXPOSURE (doys)
28 (3
z
3
20 1
Fig. 1. Median body burden of cadmium, with 95% C.I. 0
of the median, during a 28-d exposure to a nominal con-
centration of 40 mg kg- cadmium for Nereis virens 3
=
c 10
(o), Glycera dibranchiata ( A ) and Nephtys caeca (0). m
Day 0 and control cadmium tissue concentrations did not 0
exceed 1.8 mg kg-

ied and anchored into the sediment (Stage I), and

C
*
burial of the rest of the body followed (Stage 11).
Differences in Stage I burrowing times between
cadmium-exposed and unexposed worms were sta-
20
T *
- T
tistically significant ( p < 0.05) in 4 of 15 compar- 10
isons (3 species x 5 times; Fig. 2). At the initiation
of the experiment, Stage I burrowing into contam-
inated sediment was significantly faster for N. 0
virens and significantly slower for G. dibranchiata. O 7 14 21 28
Stage I burrowing by cadmium-exposed N. caeca
was significantly slower at 14 and 28 d. Differences CADMIUM EXPOSURE (days)
in mean Stage I burrowing times that were statisti- Fig. 2. Median burrowing time of head (Stage I), with
cally significant ranged from 3.2 to 11.9 s. The 95% C.1. of the median, for Nereis virens (A), Glycera
only significant difference among 15 comparisons dibranchiata (B) and Nephtys caeca (C) when placed in
uncontaminated sediment after the indicated exposure.
of Stage I1 burrowing rates was the slower burrow- Star indicates significant difference between treatment
ing of G. dibranchiata into contaminated sediment and control.
at 0 d ( p < 0.01; Fig. 3). The magnitude of this
difference was 0.86 mm s-.
posed animals ingested 100% and unexposed ani-
Effect of cadmium on feeding mals 90% of the available prey ( p > 0.05).
of G. dibranchiata
After 28 d, cadmium-exposed and unexposed DISCUSSION
G. dibranchiata presented with live E. mucronata Although some statistically significant differ-
showed no significant differences in feeding. Ex- ences in burrowing in the presence of cadmium-
 Effects of cadmium on polychaete behavior 591

contaminated sediment were observed in this study,

these differences were minor and appear to have
Cadmiumto
fpposed A little ecological relevance. In most instances, bur-
rowing time or rate was altered only by a matter
4.0
0 Not exposed of seconds. Burrowing was sometimes faster (e.g.,
N . virens at 0 d) and sometimes slower (e.g., G.
T dibranchiutu at 0 d) into contaminated sediment.
2 .o Exposure to cadmium also did not affect feeding
in G. dibrunchiata. This is perhaps more signifi-
cant than the lack of effect on burrowing, since
feeding involves a more complex integration of
0 sensory and behavioral information.
Although we found only minor behavioral al-
terations associated with cadmium exposure, we
6
T r
do not imply that such an exposure leaves a spe-
cies totally unaffected. Sublethal effects have been
demonstrated by other investigators. For example,
4.0 in examining sublethal responses of N.virens to
cadmium in seawater (10 and 40 mg L-I), Carr
and Neff [ 5 ] found that exposed worms showed
2 .o significant changes in coelomic fluid glucose levels
and a depletion in glycogen reserves. They sug-
gested that the degree of depletion, which depended
0 upon the cadmium concentration and length of
exposure, would be indicative of the animal's level
c of resistance to stress.
Cadmium uptake varied among the three poly-
chaete species we tested (Fig. 1). N. virens accumu-
lated the highest concentration of cadmium after
10.0 28 d (319 mg kgg'). Cadmium body burdens were
much less in G. dibranchiutu (120 mg kg-') and

1
lowest in N. caeca (39 mg kg-'). The low body
8.0 burden of cadmium in N. caeca may be explained
by the relatively low concentration of cadmium in
the interstitial water during the exposure experi-
6.0 ments with this species (Table 1). However, N.
virens was exposed to substantially lower cadmium
concentrations in sediment and interstitial water
4.0 but accumulated much more cadmium than did G.
dibrunchiata. These differences in bioaccumulation
. .:_.
may therefore reflect inherent biological differ-
ences between N. virens and G. dibrunchiatu.
2.o Some polychaetes are able to accumulate large
quantities of cadmium. Nereis diversicolor and N.
virens accumulated 1,860 and 1,450 mg kg-' , re-
0 I spectively, after chronic (33 to 54 d) exposures to
7 14 21 28
0 about 10 mg L-' cadmium in seawater [4,5]. In
CADMIUM EXPOSURE (days) the present study, N. virens survived, accumulated
319 mg kg-' cadmium and showed no behavioral
Fig. 3. Median burrowing rate (mm sgl) of body seg- dysfunction when exposed for 28 d to sediment
ments other than the head (Stage II), with 95% C.1. of containing about 40 mg kg-' cadmium. A sedi-
the median, for Nereis virens (A), Glycera dibranchiata ment cadmium concentration of 40 mg kg-' is
(B) and Nephtys caeca ( C ) when placed in uncontami-
nated sediment after the indicated exposure. Star indi- well within the range of contamination of the
cates significant difference between treatment and marine environment [l I]. An abundant polychaete
control. fauna, including Nereis procera and Nephtys cor-
592 B. L. OLLAET
nuta franciscana, was reported at a site on the
Palos Verdes Shelf, California, where the sediment
cadmium concentration was 32 mg kg- [12]. The 7.
potential for bioaccumulation and survival of poly-
chaetes in highly contaminated areas indicates that
species like N. virens may be an amplified source
of cadmium for the crustaceans, fish and birds 8.
that prey upon them [13-161.
Differences in uptake levels among species may
in part be due to different feeding habits. N. virens
is categorized as an omnivore [17,18] and ingests 9.
sediment surface layers, including items such as
detritus, filamentous and thallous algae, and even
sediment particles, all of which may be contami-
nated with cadmium. N . caeca has been catego-
rized as a carnivore [ 18,191 and therefore is less
likely to ingest cadmium-contaminated food items
in a study such as this. The feeding habits of G. 10.
dibranchiata are less well understood, although it
is apparent that the species is carnivorous (as in the 11.
present study) and can also be omnivorous [17,18].
Uptake of cadmium by N. virens in the present
study did not appear to be linear, in contrast to
what has been found in previous studies [2,5].Since
the amount of cadmium taken up is proportional 12.
to the amount available [2,4], the decrease in con-
centration of cadmium in the aqueous phase (espe-
cially the interstitial water) may be responsible for
the nonlinear pattern of cadmium bioaccumulation. 13.
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