HydrobiohJ,q, ia 304: 197-207, 1995.

197
@ 1995 Kluwer Academic Publishers. Printed in Belgium.

Geochemical partitioning and bioavailability of copper to aquatic plants in

an artificial oxide-organic sediment

L. A. Warren I, E M. Outridge 2 & A. R Z i m m e r m a n 1

IDepartment ofZoolog3; UniversiO, of Toronto, Toronto, Ontario M5S 1A 1, Canada
2Environmental and Resource Studies Program, Trent UniversiO; Peterborough, Ontario K9J 7B8, Canada

Received 4 August 1993; in revised form 24 February 1994; accepted 2I April 1994

Key words." copper, bioavailability, toxicity, sediments, partitioning, macrophytes

Abstract

This study investigated the effects of competition between binding substrates (organic matter and iron oxide) and
between metals (cadmium and copper), on the partitioning of sedimentary copper and its subsequent bioavailability
to an aquatic plant. Organic matter and a synthesized iron oxide, ferrihydrite, were added singly and in combination
to a series of sand sediments, which were then dosed with environmentally realistic concentrations of cadmium and
copper and planted with rice, O0'za sativa.
Organic matter controlled copper partitioning and bioavailability, whereas the synthetic ferrihydrite bound
negligible amounts of either metal, even in the absence of organic matter. As organic matter concentrations
increased, operationally-defined leachable copper decreased, organic-associated copper increased and the survival
of rice plants improved in an approximately linear fashion. At a nominal starting copper concentration of 5.8 F~g g
dry wt -I , plant survival after four weeks averaged 0-8% in sediments without organic matter, 25% in a sediment
containing 0.18% organic matter and 58% in a sediment containing 0.36% organic matter. These results suggest
that organic-associated forms of copper are unavailable to plants, and that the operational definition of 'leachable'
copper (extracted with dilute ammonium acetate) adequately represents the species of copper that is (are) available
to plants. Our study using a well-characterized artificial sediment supports the copper fractionation patterns and
correlations between copper partitioning and bioavailability reported from the heterogeneous, poorly characterized
sediments of natural lake and river sediments.

Introduction ligands (Cahnano et al., 1988; Fulghum et al., 1988;

Aquatic systems constitute the largest sink for heavy
metals mobilized by anthropogenic activities (Nriagu
& Pacyna 1988). Currently, a prevalent paradigm for
metal toxicology and biogeochemistry is that geo-
chemical partitioning between solid (sediment) and
solution phases is the key factor controlling the avail-
ability, and hence toxicity, of metals to biota in aquatic
environments (Campbell et al., 1988; Luoma, 1989).
Furthex'more, the partitioning of metals in sediments
(i.e. their distribution among different geochemical
substrates), is influenced by environmental factors
such as pH and Eh, the abundance of organic and inor-
ganic materials (clays and hydrous metal oxides), and
the concentrations of competing cations and solution
Tessier et al., 1985).
Beyond these broad generalizations, however, a
number of specific questions remain unresolved. One
issue concerns the relative sorptive affinities of particu-
late organic matter and metal oxides for different trace
metals. For example, in oxidized, organic-rich natural
sediments, copper is usually most strongly associat-
ed with organic matter (Warren & Zimmerman, 1993;
Chen et at., 1989; Tessier et al., 1980), although this
association may be less important in sediments that are
relatively high in iron oxides and low in organic mat-
ter (Luoma & Bryan, 1981; Luoma, 1986). In contrast,
metals such as cadmium tend to be associated only
weakly with organic matter, favouring the carbonate
and 'exchangeable' fractions (Warren & Zimmerman,
 198
1993; Pardo et al., 1990; Tessier et al., 1980), or metal
oxides (Reuther et al., 1981 ).
Another primary area of interest is the nature of the
relationship between metal partitioning and bioavail-
ability. Key questions include the identity of the
'bioavailable" fo,ms of metals, how they are quanti-
tatively influenced by sediment constituents, and the
most appropriate means of measuring these forms
(Luoma, 1989: Campbell & Tessier, 1989). One
approach used to address these questions has been
to test for correlations between various operationally-
defined metal fractions in sediments and either the met-
al concentrations in or toxicity to sediment-associated
biota such as macrophytes, bivalves and other benthic
invertebrates (e.g. Campbell et al., 1985; Thompson
et al., 1984: Luoma & Bryan, 1978). A positive corre-
lation between the concentrations in a given sediment
fraction and ill biota would suggest that the fraction
in question is "bioavailable'. Most studies of this type
have been carried out on natural systems, analysing
sediments and biota collected from the field.
In contrast, a second approach, which has produced
most of our current theoretical knowledge regarding
metal sorption onto solid phases, has largely involved
laboratory model systems using well-defined "artifi-
cial' particles, predominantly metal oxides (Honey-
man & Santschi, 1988). Because of the complexi-
ty of metal-particle interactions, two common fea-
tures of this type of study are single metal-substrate
experimental designs, and the use of synthetically-
derived goethite (a-FeOOH), a crystalline Fe oxide
(e.g. Ankomah, 1992; Barrow et al., 1989). In nat-
ural sediments, however, other oxides such as ferri-
hydrite and lepidocrocite have been identified (Fortin
et al., 1993: Tessier et al., 1993), which are more
amorphous than goethite and consequently may have
quite different sorption characteristics (Schwertmann
& Cornell, 1991). Natural systems may also contain
multiple binding substrates in potential competition for
metals. Thus, the degree to which the laboratory-based
literature has direct relevance to questions of metal
bioavailability in natural systems remains unclear. Rel-
atively few studies of metal-particulate associations in
natural systems (e.g. Tessier et al., 1993, 1989, 1985,
1980; Warren & Zimmerman, 1994; Bendell-Young
& Harvey, 1992) are available for comparison with
laboratory-derived literature.
This study aims to link these two different
approaches, by employing a well defined artificial sed-
inaent that has environmentally relevant characteris-
tics, namely, competing binding substrates and metal
cations, and realistic concentrations of metals. Tile
objective of our laboratory-based study is two-fold.
First, we examine the geochemical partitioning of cop-
per in a series of sandy sediments containing varying
proportions of a synthesized iron oxide (ferrihydrite)
and organic matter (peat). The effect of another cation,
cadmium, on the partitioning of copper was also inves-
tigated. Second, we relate tile observed differences in
partitioning to the availability of copper to an aquatic
macrophyte, O0'za sativa, that was grown in the sedi-
ments. Specifically, we test the hypothesis that increas-
ing amounts of organic matter in sediments will lower
the bioavailability of copper to the macrophyte.
Materials and methods
E x p e r i m e n t a l design
The experimental design was an unbalanced two-way
factorial analysis of variance (Table 1), set up as a
sand culture experiment (Hewitt, 1966). Two different
metal treatments, Cu alone or a mixture of Cu + Cd,
were added to each of 4 sediment types: the sand sed-
iment, 700 g of fine silica sand (SAND): oxide sedi-
ment, 2.5 g of ferrihydrite added to 700 g sand (OX);
organic sediment, 2.5 g of peat added to 700 g sand
(OM); and mixed sediment, 1.25 g each of ferrihydrite
and peat added to 700 g sand (MIX). To account for
any differences in partitioning due to the presence of
plants, a sediment control (Cd +Cu treatment with no
plants) was included for each sediment type. Similarly,
to account for any differences in plant growth due to
sediment type, a plant control (no metal treatment) was
also included for each sediment type.
Ferrihydrite binding capacity was assumed to be
0.005 tool metal per tool Fe at a pH of 6.5 to 7,5
(Dzombak & Morel, [990), so that 2.5 g of ferrihydrite
should theoretically be able to complex 2.6 × 10 - 4
moles of metal ions. An initial experiment was car-
ried out with the peat to determine its binding capacity.
A 200 rnl volume of 5 mg 1- I Cu was added to a
series of beakers containing fiom 0.25 to 3.0 g of peat,
and porewater Cu concentrations were measured after
one week. This experiment revealed that 2.5 g of peat
was able to complex 2 x 10 - 4 moles of Cu, rough-
ly equivalent to that for the same mass of Fe oxide.
Both the Cu and the Cu + Cd treatments employed the
same total molar concentration of metal (8.9 × 10 - s
M) in each sediment type. In the Cu + Cd treatment,
this was achieved by adding 3.12 × 10 -5 moles each
 199

Oxide and organic matter in amounts appropriate

Table 1. Experimental design
for each sediment type were individually mixed with
Sediment type 700 g of air dried silica sand in plastic bags. The
Metal Sand Oxide Mixed Organic metal treatments were added in solution (50 ml vol-
Treatments Matter ume diluted from BDH 1000 ppm Atomic Absorption
Standard), and the sediment again thoroughly mixed
Sediment
before being transferred to 100 mm Standard plastic
control
(no plants, 11=4 n=4 n=4 n=4
plant pots. Each pot was acid washed prior to use, and
Cu+Cd)
contained an acid-washed circle of nitex mesh (I #m
pore size) at the bottom to retain sediments and allow
Plant for water movement into the pots. The pots were then
control n = 6 n = 6 n= 6 n= 6 kept immersed in deionized ultrapure water buffered
(no metal ) to pH 6.5 to allow for development of a partitioning
equilibrium over a two week period. An initial sedi-
Cu 5.8 Fzg g - 1 n = 4 11 = 4 n= 4 n= 4 ment sample was collected for fractionation prior to
rice seedlings being planted in the pots at the start
Cu + Cd of the experiment (see below). Samples were collect-
5 # g g - I Cd + ed from three replicates from each of the sand sed-
2 . 9 / z g g - t Cu n=4 n=4 n=4 n=4 iments (SAND column, Table 1), and from all four
sediment types in the Cu + Cd treatment (bottom row,
Table I ).

of Cu and Cd to the 700 g of sediment in each pot,
giving nominal concentrations of 5 #g g dry weight-I
Cd and 2.9 pg g dry wt - I Cu, while in the Cu treat-
ment, 6.23 x 10 -5 moles of Cu was added (giving a
concentration of 5.8 Ftg g - l ) . The metal concentra-
tions added approximately an order of magnitude less
cations than could theoretically be complexed by the
number of particulate binding sites available, thus pro-
moting competition between geochemical fractions for
metal ions. In addition, the metal concentrations are in
the range reported for sediments in lakes near Sudbury,
Ontario (Tessier et al., 1993, 1985), with the Cd con-
centration well below the threshold toxicity level of
30 Itg g dry w t - I reported for rice (Muramoto et al.,
1990).
Sediment preparation
Ferrihydrite was prepared by precipitation with alkali
following the protocol of Schwertmann and Cornell
(1991). Briefly, 1M KOH was added to Fe(NO3)3 •
9H20 dissolved in distilled water, until the pH of the
solution was between 7 and 8. The resulting slurry was
then centrifuged (12000 × g , 10 minutes) to remove
the supernatant, dialyzed over 2-3 days to remove elec-
trolytes, and freeze dried. The peat (Canada Brand,
Annapolis Valley Peat Moss Co., Nova Scotia) was
sieved to retain only particles less than 2 mm in diam-
eter.
Plant material and culture
Seedlings of rice (Oryza sativa var. M 101 ) were grown
in a soil/peat mixture under combined natural and flu-
orescent light in a greenhouse for five weeks, at which
time they were approximately 20 cm high. The plants
were carefully separated from the soil, and the shoots
and roots trimmed to 5 cm and I cm respectively. Six
plants were collected for initial dry wt and copper and
cadmium analyses. Three seedlings were transplant-
ed into each of the pots containing the experimental
sediments, and placed in the greenhouse under natu-
ral light supplemented by fluorescent lights on a 14:10
light:dark cycle. Each pot was placed in a larger con-
tainer that acted as a water reservoir, ensuring the sedi-
ment remained saturated during the experiment. Ambi-
ent temperatures in the greenhouse varied daily from
approximately 23 to 35 °C.
At intervals of four days, each pot received 40 ml
of nutrient medium in its external water reservoir. The
nutrient was Peter's Professional 20/20/20 Medium,
added at a rate of 5 g l-I distilled water, and was
adjusted to pH 6.5 with 1 M HNO3 and NaOH. Dis-
tilled water that was adjusted to pH 6.5 was added as
necessary to maintain sediment saturation. Plant mor-
tality was noted every second day, and dead plants
removed.
The experiment was terminated after 28 days, when
approximately 10 g of sediment for metal fractionation
200
analysis was sampled from the upper three cm of each
pot. All plants were then carefully excavated, washed
with distilled water to remove adhering sediment par-
ticles, and dried at 60 °C overnight. After cooling,
they were weighed, and digested with hot concentrat-
ed HNO3. The digestate was cooled and made up to
constant volume with deionized-distilled water.
Sediment extraction
All glassware, centrifuge tubes, vials and pipettes were
acid washed in 10% HNO3 and rinsed with deionized
water five times prior to use. A sequential extraction
procedure, modified from Tessier et al. (1979), was
used to extract metals in four operationally defined
phases: leachable (exchangeable and carbonate associ-
ated: 1M NH4OAc, room temperature for 5 hours with
continuous agitation); reducible (oxide associated;
0.04M NH~OH.HCL in 25% HOAc, heated to 96 °C.,
with occasional agitation for 6 hours); oxidizable
(organic associated; 0.02M HNO3, 30%H202, heat-
ed to 85 °C with occasional agitation for 3 hours); and
residual (resistate mineral phases; 4: l HNO3 + HCIO4
digestion, extracted with 4:1 deionized water+ warm
6M HC1). A 20:1 ratio of reagents to dry weight of
sediment was used (see Warren & Zimmerman (1993)
for a complete set of reagents and conditions).
The supernatants from each extraction step were
analyzed for Cu and Cd using flame atomic absorption
spectroscopy (Laboratory Instruments Spectropho-
tometer, Model 351 ). Blanks and NBS reference mate-
rial (NBS 2704, Buffalo River Sediment) were also
run through the extraction; the total concentrations of
metal recovered from the latter samples were within
the certified ranges.
Statistical procedure
Data on metal concentrations were log transformed,
and proportional metals and plant survival data arc-
sine transformed, prior to statistical analysis. Plant
survival was analysed by analysis of variance with
repeated measures, followed by Duncan's multi-range
test for differences between sediment types. All other
data were analysed by ANOVA followed by Duncan's
test for differences between sediment types and metal
treatments. Data in figures and the text are shown as
mean+standard error. Significance levels in the text
are at p<0.05 unless stated otherwise.
Results
Initial concentrations and partitioning of metals
Tile total Cu concentration in the plant control sand
sediment (i.e. no added metal) at the start of the exper-
iment was 0.67-1-0.03 ttg g dry wt - I . By compari-
son, the sand sediment of the Cu treatment contained
1.9 #g g - i Cu (Fig. la), which was markedly low-
er than expected (nominally 5.8 #g g - i ) . Mean Cu
concentrations recovered initially from the C u + C d
treatment ranged from 2.26 to 3.10 #g g - I , which
were close to the nominal concentration of 2.9 #g g-~,
and showed no significant differences between sedi-
ment types (/?>0.88; Fig. lb). Therefore, the relative
proportions of Cu in each geochemical fraction were
compared across all four sediment types in the Cu + Cd
treatment. The leachable and oxidizable forms of Cu
dominated in different sediment types. In the sand sed-
iment, the proportion of leachable Cu (68%) was sig-
nificantly higher than in other sediment types, while
the organic sediment (OM) had a significantly high-
er proportion of oxidizable, i.e. organic-associated,
Cu (69%). Surprisingly, reducible Cu (i.e. oxide-
associated) was significantly higher in the mixed (29%)
than in the oxide sediment (12%), which contained
about the same relatively small amount of reducible
Cu as the sand and organic sediments. The fractiona-
tion patterns in the sand sediment were not significant-
ly different between the Cu and Cu +Cd treatments
(cf. Figs la, lb; p>0.10), suggesting that the presence
of Cd had no effect on the partitioning of Cu.
The total concentrations of Cd recovered initially
in the Cu +Cd treatment were similar to the nominal
level of 5 #g ~o.--I , and showed no significant differ-
ences between sediment types (p>0.70; Fig. lc). In
terms of fractionation, the leachable form of Cd was
dominant. Similarly as for Cu, the sand sediment had
a significantly higher leachable fraction of Cd (91%)
than other sediment types, while the OM sediment had
the lowest (61%). Reducible Cd was significantly high-
er in the oxide, mixed and organic sediments compared
to sand.
Plant survival, growth and metal accumulation
The concentrations of Cu and Cd used in these treat-
ments were far more toxic to the rice plants than was
expected, based on previous studies with rice. Both Cu
and Cu + Cd treatments resulted in rapid mortality of
plants in most sediment types, with <20% survival by
 201

c,, (o) ,oo,\_ ..... ...............................................

(Cu Treatment)

.............................,,:_ ..................
i %
d~ S
{;~f,:3 RESIDdAL
U
OX~DIZASIE R
I I REOUCIBLE V
LEACHABLE I
1
V
O
R 40 . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .

0 S
H
I
P

Cu (b)
(Ou+Od Treatment)

3 0 5 10 15 20 25 30
7 ELAPSED TIME (DAYS)
Cu

- - SAND o OX o MiX - ~ - OM
1 Fig. 2. Plant survivorship in the Cu treatment.

0 100~

od (c) 80
(Cu+Od Treatment)
%

S
U 60
q~ R
V
I
V
O
R 40
S
H
t
SAND OX MIX Old P

Fig. 1. The total concentrations and proportions in each extracted 20

fraction of (a) Cu in the Cu treatment, (b) Cu in the Cu +Cd treat-
ment, and (c) Cd in the Cu + Cd treatment, at the start of the exper-
iment. Concentrations are shown as mean totals-t-standard error;
proportional data are shown as means, represented within each bar. 0
0 5 10 15 20 25 30
ELAPSED TIME (DAYS)
Cu + Cd
the end of the experiment (Figs 2, 3). The majority of Fig. 3. Plant survivorship in the Cu + Cd treatment. Symbols as in
plants in the plant control survived (Fig. 4), although Fig. 2.
mortality (22%) in the oxide sediment was significant-
ly greater than in other sediments.
The exception to the pattern of high mortality was (organic-oxide) sediment also had significantly elevat-
in Cu treatments with sediments containing organic ed survival compared with the oxide sediment. The
matter (Fig. 2). Plants grown in the organic sediment differences in survival between sediment types were
exhibited significantly higher survival than in any oth- roughly proportional to their levels of organic matter,
er sediment throughout the experiment. The mixed since survival averaged 58% in OM pots containing
202

100'
Table 2. Finalbiomass and copper content of plants surviving to end
of the experiment+ .

80 ............................................ Metal Sediment

type
treatments Sand Oxide Mixture Organic
%
manet
S
U 60 .....................................................
A) Biomass (g dry wt)x
R
V
I Plant 0.024 0.022 0.033 0.029
V
0 Control -4-0.003 + 0.005 4- 0.022 + 0.011
R 40 . . . . . . . . . . . . . . . . . . . . . .
S
H
1
Copper 0.036 - 0.036 0.045
P (n = 1) 4- 0.009 4- 0.037
20 . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .
Copper + - 0.032
Cadmium (n = I)

0 I I I t I I
B) Copper content (#g g dry wt- I)~
0 5 10 15 20 25 30
ELAPSED TIME (DAYS)
Plant 79 q-- 1l A 90 4- 35A 74 4- 37A 38 4- l0 B
CONTROL
Control
Fig. 4. Plantsurvivorshipin the control treatment(no added metal).
Symbols as in Fig. 2.
Copper I. 195 165 4- 81 328 4- 134
01 = 1)

Copper + - 124
2.5 g (0.36%) of organic matter, 25% in MIX pots that
Cadmium (n = I)
had 1.25 g (0.18%) organic matter and 0 - 8 % in the
OX and SAND sediments that contained no organic
matter. +: data shown as mean 4- standard deviation
In terms of plant growth, there was no significant =: initial biomass was 0.014 4- 0.002 g dry wt (n = 6)
difference between sediment types in any metal treat- (~:initial copper content of plants was 36 4- 12 ttg g dry wt- J
Different superscripted letters indicate differences between sediment
ment (Table 2a.). However, plants that survived in the types significantat p<0.01.
Cu treatment produced markedly more biomass than
those in the control. Plants in the Cu-treated sediments
grew an average of 2.6- to 3.2-fold during the study
period, compared to only 1.7- to 2.4-fold in the con- Changes in metal partitioning during the
trol, suggesting that Cu deficiency was limiting growth experiment
in the control treatment. This suggestion is supported
by the Cu content data on the plants. The Cu-treatment Sediment samples taken at the end of the experiment
plants contained significantly higher Cu concentrations enable changes in the sedimentary partitioning and
than control plants (Table 2b), which no doubt reflects concentrations of metals during the experiment to be
the higher Cu concentrations in the treated sediments. identified. A c o m m o n feature of oxide sediments in all
An interesting result from the plant control was that metal treatments was a loss of most of the added metal
plants grown in the organic sediment exhibited sig- from the pots. In the Cu + Cd treatment, for example,
nificantly lower Cu concentrations than those in other this loss amounted to approximately 63% (1.13 g) of
sediments (p<0.01). In fact, the plants in OM sedi- the Cu and 85% (2.53 g) of the Cd present at the start
ment contained virtually the same concentration of Cu of the experiment. The mixed oxide-organic sediment
(38/~g g - i ) as seedlings at the start of the experiment showed smaller losses of Cu and Cd, and the sand
(36 #g g-X), while plants in other control sediments sediment exhibited similar losses of Cd. Presumably,
more than doubled their Cu contents during the exper- this metal flowed out of the sediments into the external
iment. reservoirs. The following discussion will therefore be
 203

confined to those treatments which did not show sig-

nificant losses of total metal during the experiment.
In the Cu-treated sand sediment, the total Cu con- 4
v///J/////.4(Cu Cu
Treatment)

centration increased slightly during the experiment

27 70
(p>0.18, Fig. 5a cf. Fig. la), probably due to Cu T 3
added in the nutrient medium. Across sediment types,
the proportion of leachable Cu decreased, and oxidiz- 2
able Cu increased in an approximately linear fashion,
as the amount of organic matter increased (Fig. 5a). 1
The organic sediment had a significantly lower pro-
portion of leachable Cu (9%) than the sand (35%), and o

a significantly higher proportion of organic-associated

Cu (70%) than either the mixed (47%) or sand (27%)
sediments at the end of the study. ou (b)
(Cu+Cd Treatment)
In the Cu +Cd treatment, total recoverable con-
centrations of Cu again increased significantly in the
organic sediment, and decreased significantly in the
T
oxide sediment, but did not change appreciably in the c~
mixed or sand sediments (Fig. 5b). In terms of parti- ::L
tioning, a decrease in leachable Cu was observed in
the mixed sediment during the experiment (leachable
Cu initially 21%, final 8%; p<0.01). In the sand sed- 10

iment, reducible Cu increased during the experiment ;;ii~ii;iii

(initially 12%, final 34%; p<0.01) while there was no

change in the proportion of leachable forms. The losses
10
of total Cd observed in the mixed and sand sediments 9 Od (C)
over the study period were due to significant decreases 8 (Cu+Cd Treatment)
in leachable forms (p<0.01; Fig. 5c, cf. Fig. lc). No 7
significant difference was observed between initial and T 6 7""
final partitioning of Cd in the organic sediment. 5 a4
Cr~
::L 4

Discussion

Copper fractionation in these artificial laboratory sed- o

iments was strongly influenced by the presence of
organic matter, with the fraction of oxidizable (organic-
associated) Cu increasing proportionally with organic
matter content. Our data are consistent with the strong
association of Cu with organic matter that is well
documented across a variety of natural aquatic sedi-
ments: lake sediments (Pardo et al., 1990; Bendell-
Young & Harvey, 1992; Chen et al., 1989); fluvial
suspended particulate matter (Warren & Zimmerman,
1993); river sediments (Tessier et al., 1980); and estu-
arine sediments (Luoma & Bryan, 1981). Leachable
forms of Cu decreased as oxidizable Cu increased in
this experiment, while the reducible and residual frac-
tions remained relatively constant, suggesting a con-
version from leachable (more loosely associated) to
SAND OX MIX OM
Fig. 5. The total concentrations and proportions in each extracted
fractionof(a) Cu in the Cu treatment, (b) Cu in the Cu + Cd treatment,
and (c) Cd in the Cu + Cd treatment, at the end of the experiment.
Legend as in Fig. 1.
oxidizable-associated Cu in the presence of organic
matter.
These changes in fractionation among sediment
types were associated with changes in copper bioavail-
ability and toxicity. The decrease in leachable Cu and
increase in oxidizable Cu was accompanied by signifi-
cantly improved survival of the rice plants growing in
the organic sediment. These results lead to two conclu-
sions. First, organic-associated Cu was less bioavail-
able to the plants than leachable Cu. This finding was
 204
supported by data from tile control treatment in which
plants grown in organic sediments accumulated signif-
icantly less Cu than plants in sediments with lower or
no organic matter. Similarly, Campbell et al. (1985)
reported a somewhat lower correlation between oxi-
dizable Cu in river sediments and Cu in macrophyte
tissues, than with extractable Cu, again indicating a
lower availability of oxidizable forms of Cu. The sec-
ond conclusion is that the operational definition of
'leachable' Cu employed here (i.e. dilute ammoni-
um acetate extraction) provides a good approximation
of the bioavailable forms of Cu. It appears that vari-
ous extraction schemes may adequately represent the
bioavailable fraction of sedimentary Cu, since field
studies of the fractionation of Cu in underlying sed-
iments and concentrations in tissues of Nuphar lutea
(Aulio, 1980) and Nuphar variegatum (Campbell etal.,
1985) found similarly good agreement using different
reagents to define the 'extractable' or 'leachable' frac-
tions.
We saw no consistent evidence that the
synthetically-derived iron oxide, ferrihydrite, bound
Cu or Cd to an appreciable degree. The mixed organic-
oxide sediment was found to contain a larger propor-
tion of oxide-associated Cu than the organic (oxide-
fiee) sediment, suggesting that competition for Cu
might have occurred between organic matter and oxide
in the mixed sediment. However, the oxide sediment
contained negligible amounts of oxide-associated Cu,
whereas the majority of Cu in the organic sediment
was organic-associated. This indicates that under the
conditions of the experiment, organic matter was a
more effective sink for Cu than the synthetic oxide
on a unit weight basis, corroborating studies of Cu
partitioning in natural systems in which both organic
matter and iron oxide were present (Warren & Zimmer-
man, 1993; Bendell-Young & Harvey, 1992). It should
be noted that both the pH of the experimental medium
(pH 6.5) and the type of organic matter used here (peat,
a refractory organic substance) would favour the asso-
ciation of Cu with organic matter (Inskeep & Baham,
1983). Our findings differ from those of Luoma &
Bryan (1981) and Luoma (1986) who reported that
organic-Cu associations in estuarine sediments were
less dominant in the presence of increasing amounts
of iron oxides. While a definite explanation is elusive,
the discrepancy between studies may reside in differ-
ences in ambient pH, the type (natural versus synthetic)
and concentrations of iron oxides in each system, or
changes in organic behaviour at the relatively higher
ionic strength of estuarine waters.
Other potential influences on the partitioning of
Cu, i.e. Cd as a competing cation, and the presence of
aquatic plants, were found to be negligible. There were
no significant differences in partitioning between the
Cu + Cd (with plants) and sediment control (Cu + Cd
without plants) treatments. It may be that plants are
only a factor in reduced sediments, in which plant
roots can create an oxidized mizosphere that alters
metal partitioning relative to the surrounding reducing
conditions (Crowder et al., 1987; Crowder & Mac-
fie, 1986). Here, the oxidized nature of the sediments
would preclude an effect from the plants. While sedi-
ment Eh was not measured, we are reasonably certain
that the sand-based sediments remained oxidized, as
visual examination of sediment profiles at the end of
the experiment revealed no black precipitate indicating
the presence of iron sulphides i.e. reduced conditions.
The absence of Cd competition for Cu binding sites was
indicated by the consistent similarities in Cu fraction-
ation between the Cu and tile Cu + Cd treatments with
the sand sediment at the start of the study, and across
all sediment types at the conclusion of the study. Our
findings are in accord with tile concept of heterogene-
ity of solid binding sites suggested by Benjamin and
Leckie (1981), who showed that particulate binding
sites that have a high affinity for Cu are not preferred
for Cd, and thus minimal competition between the two
metals could be expected.
An unexpected result was the 'loss' of recoverable
Cu and Cd from the oxide sediment during the experi-
ment. Sand culture experiments such as these are rou-
tinely used in plant nutrition and phytotoxicity studies
(Hewitt, 1966), and no such loss of Cu occurred in the
sand or organic sediments during this study, suggesting
an effect associated with the presence of the synthetic
ferrihydrite. Strangely, the losses did not occur during
the two week sediment equilibration period prior to
the experiment. Scanning electron microscope (SEM)
images of the oxide sediment at the start of the exper-
iment indicated a relatively amorphous coating of Fe
oxide on sand particles (Fig. 6). We hypothesize that
this coating may account for the loss of metal, if it
prevented metals from sorbing to sand grains. As dis-
cussed, the oxide was ineffective at binding Cu or
Cd from solution, in apparent contrast to naturally-
occurring metal oxides. Diagenesis of the oxide was
also apparent, because SEM images taken one month
after the experiment concluded revealed that the oxide
no longer coated sand particles as amorphous mate-
rial, but was more crystalline in appearance (Fig. 7).
These age-related changes, which have been report-
 205

• t

/
\

, !

Fig. 7. SEM images of the OX sediment taken one month after the
Fig. 6. SEM images of the OX sediment after the two week
experiment concluded, i.e. oxides were kept in an aqueous state for
equilibration period, prior to the introduction of the plants. A sand
8 weeks. A sand grain coated by Fe oxide appears in the middle of
grain coated by Fe oxides can be seen in the middle of (a), and this
(a), and this same sand grain shown at higher magnification in (b).
same sand grain shown at higher magnification in (b). Fe oxides
Fe oxides appear more crystalline in comparison to the images in
appear as a relatively amorphous coating on the sand grains.
Fig. 6.
 206
ed p r e v i o u s l y in h y d r o u s ferric o x i d e s , are e n h a n c e d
b y h i g h e r t e m p e r a t u r e s ( 2 0 - 3 0 °C; D z o m b a k & M o r e l ,
1990). T h e e x p e r i m e n t a l c o n d i t i o n s o f this s t u d y w h i c h
i n c l u d e d a m b i e n t t e m p e r a t u r e s o f 23 to 35 ° C w o u l d
f a v o u r the a g i n g p r o c e s s . T h e a p p a r e n t d i a g e n e s i s o f
the F e o x i d e p r e c l u d e d t e s t i n g t h e a b o v e h y p o t h e s i s in
a later e x p e r i m e n t b e c a u s e t h e s o r p t i v e c a p a c i t y o f t h e
o x i d e c o u l d h a v e also a l t e r e d ( S c h w e r t m a n n & C o r -
nell, 1991; D z o m b a k & M o r e l , 1990). T h u s , f u r t h e r
w o r k with f e r r i h y d r i t e , w h i c h m a y b e a m o r e e n v i r o n -
m e n t a l l y realistic o x i d e than g o e t h i t e (Tessier et al.,
1993; F o r t i n et al., 1993) s h o u l d take into a c c o u n t
d i a g e n e t i c e f f e c t s that m a y alter t h e o x i d e ' s b i n d i n g
characteristics.
Acknowledgments
This work was funded through an NSERC Operating
g r a n t a n d a R A C g r a n t f r o m the O n t a r i o M i n i s t r y o f
the E n v i r o n m e n t to A P Z ; a g r a n t f r o m the C a n a d i a n
W i l d l i f e F o u n d a t i o n to A P Z a n d P M O ; a n d an N S E R C
P o s t d o c t o r a l S c h o l a r s h i p to P M O . W e t h a n k D r G r a n t
Ferris, D e p a r t m e n t o f g e o l o g y , U n i v e r s i t y o f T o r o n t o
for c o m m e n t s o n o x i d e d i a g e n e s i s , a n d M r Eric Lin,
D e p a r t m e n t o f Z o o l o g y , U n i v e r s i t y o f T o r o n t o , for his
assistance with SEM work.
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