ARTICLE IN PRESS

Ecotoxicology and Environmental Safety 63 (2006) 175–188

www.elsevier.com/locate/ecoenv

Rapid communication

Food chain transfer of selenium in lentic and lotic habitats of a western

Canadian watershed
Patricia L. Orra,, Karin R. Guiguerb, Cynthia K. Russela
a
Minnow Environmental Inc., 6800 Kitimat Road, Unit 13, Mississauga, Ont., Canada L5N 5M1
b
Franz Environmental Inc., 8177 Torbram Road, Brampton, Ont., Canada L6T 5C5
Received 8 August 2005; received in revised form 7 September 2005; accepted 9 September 2005
Available online 4 November 2005

Abstract

Selenium (Se) is an essential micronutrient, exhibiting a narrow margin between nutritionally optimal and potentially toxic
concentrations. Egg-laying vertebrates at the top of aquatic food chains are most at risk in environments with elevated aqueous Se
concentrations. The Elk River watershed in British Columbia, Canada receives effluents containing Se from five coal mine operations.
This study tested three hypotheses that might account for higher Se concentrations in fish from lentic compared to lotic habitats in the
watershed: (1) enhanced uptake by aquatic primary producers, (2) longer food chain length, or (3) greater food web accumulation
through sediment–detrital pathways. Stable isotope and Se concentration data demonstrated that Se concentrations in aquatic primary
producers and food chain lengths were comparable in lentic and lotic habitats. Enhanced formation of organoselenium and subsequent
uptake and cycling via sediment–detrital pathways likely account for higher fish tissue Se concentrations in lentic than in lotic areas.
Crown Copyright r 2005 Published by Elsevier Inc. All rights reserved.

Keywords: Selenium; Stable isotopes; Coal; Food chain; Biomagnification; Fish; Lentic; Lotic; River

1. Introduction
Selenium (Se) is an essential micronutrient for animals
serving beneficial metabolic functions (Arthur and Beckett,
1994), although there is a very narrow range between
nutritionally optimal and potentially toxic dietary levels for
vertebrates (Wilber, 1980; NRC, 1989). Se is much less
toxic to most plants and invertebrates than to vertebrates
and, among vertebrates, egg-layers such as birds and fish
have substantially lower thresholds for reproductive
toxicity than mammals (USDOI, 1998).
Se shares many properties in common with sulfur, which
sits above it in the periodic table (Robberecht and Van
Grieken, 1982). Se toxicity occurs when aquatic organisms
accumulate it from the water column, sediments, or food,
synthesize amino acids where the sulfur is replaced by Se,
and utilize selenoamino acids as replacements for analo-
gous amino acids in the formation of proteins (Weiss et al.,
1965; Butler and Peterson, 1967; Wrench, 1978; Bottino
Corresponding author. Fax: +1 905 567 6805.
E-mail address: porr@minnow-environmental.com (P.L. Orr).
et al., 1984; Besser et al., 1994; Maier and Knight, 1994;
Ogle and Knight, 1996). This alters the three-dimensional
structure of the proteins, resulting in impairment of
enzymatic function (Demayo et al., 1979; Alaimo et al.,
1994).
Biogeochemical cycling of Se in water bodies varies as a
function of hydrology (retention time), rates of biotic and
abiotic Se transformations, reduction and oxidation
(redox) conditions in sediments, food web structure,
species-specific differences in uptake and depuration
kinetics, and other factors (Sappington, 2002). Se may be
more readily assimilated into the food chains of slow-
flowing (lentic) water bodies than those of high-flowing
(lotic) water bodies (Lillebo et al., 1988; Canton and Van
Derveer, 1997; Lemly, 1999). Such environments are
thought to be conducive to Se accumulation, biotransfor-
mation, and remobilization through the benthic–detrital
food web and rooted plants (Lemly and Smith, 1987; Maier
and Knight, 1994; Canton and Van Derveer, 1997). By
comparison, in lotic waters, fine organic sediments such as
those produced by the deposition and decay of particulate
matter and plant and animal tissue may be rare because

0147-6513/$ - see front matter Crown Copyright r 2005 Published by Elsevier Inc. All rights reserved.
doi:10.1016/j.ecoenv.2005.09.004
 ARTICLE IN PRESS
176 P.L. Orr et al. / Ecotoxicology and Environmental Safety 63 (2006) 175–188
they are continually flushed from the system (Lemly, 1999).
Therefore, there is little opportunity for a contaminated
surface layer of sediment to develop and rooted plants are
often scarce. Furthermore, the benthic–detrital compo-
nents and associated food web of lotic environments may
play smaller roles in Se cycling, resulting in reduced
accumulation efficiency relative to that of standing water
with similar aqueous concentrations (Canton and Van
Derveer, 1997). However, no previous studies specifically
investigated Se transfer through the food webs of lotic
versus lentic areas.
The Elk River watershed, located in the extreme south-
eastern portion of British Columbia (Fig. 1), includes the
Kootenay geological formation, an area of naturally
seleniferous soils. In addition to contributions from the
local geology, five coal mines operate within the watershed
and discharge wastewaters containing Se. Elevated con-
centrations of Se have been found in water, sediment, and
aquatic biota in some areas of the watershed downstream
of the mines (McDonald and Strosher, 1998; McDonald
and Strosher, 2000; Kennedy et al., 2000; EVS, 2002a, b;
Minnow, 2003). Fish in some lentic areas downstream of
the mines were found to contain whole-body Se concentra-
tions of up to 42 mg/kg dry weight (dw; Minnow, 2003),
whereas fish in lotic areas typically showed concentrations
of less than 10 mg/kg dw (data in EVS (2002a) converted
from muscle and gonad concentrations using equations
presented by USEPA (2004)). The US Environmental
Protection Agency recently proposed a whole body
criterion of 7.91 mg Se/kg dw to protect against reproduc-
tive impacts on fish populations (USEPA, 2004).
Several hypotheses were considered potential explana-
tions that might account for greater Se concentrations in
fish and amphibian tissues in mine-exposed lentic areas of
the Elk River watershed:

Greater Se uptake by algae and aquatic macrophytes,
thus contributing to higher Se concentrations through-
out the entire food chain of lentic compared to lotic
environments. Numerous studies have shown that Se is
readily taken up from water by algae (Sandholm et al.,
1973; Nassos et al., 1980; Foe and Knight, 1986; Riedel
et al., 1991; Besser et al., 1993; Bowie et al., 1996; Herbel
et al., 2002) and aquatic plants (Allen, 1991; Ornes et al.,
1991; Herbel et al., 2002);

Greater food chain length in lentic than in lotic
environments leading to more accumulation among
the highest consumers (e.g., fish and amphibians). Stable
nitrogen isotope data were used in other studies to
identify that longer food chain length accounted for
high concentrations of persistent contaminants in
certain fish populations (Cabana and Rasmussen,
1994; Kidd et al., 1995);

Differences in the base of the food web structure that
result in different uptake pathways for Se in lentic
compared to lotic habitats. Numerous authors have
suggested that microorganisms associated with sedi-
ment–detritus may be important in food web incorpora-
tion of Se (Bender et al., 1991; Alaimo et al., 1994;
Bowie et al., 1996; Canton and Van Derveer, 1997; de
Souza et al., 1999; Hamilton and Lemly, 1999; Herbel et
al., 2002; Hamilton and Buhl, 2003).
These hypotheses were tested by collecting samples of
water, sediment, primary producers (algae and aquatic
plants), benthic invertebrates, and fish in seven lentic areas
and three lotic areas, including both reference and mine-
effluent-exposed environments in 2002 and 2003. Stable
isotopes and Se were analyzed in abiotic and biological
samples to identify the food chain pathway(s) leading to
the elevated concentrations of Se in fish tissues.
Stable isotopes of an element have the same number of
protons in the nucleus of their atoms but have a different
number of neutrons and thus different atomic mass. Stable
isotopes of carbon (13C/12C), nitrogen (15N/14N), and
sulfur (34S/32S) are most often used in environmental
studies. Results are usually expressed as delta or d
(referring to deviation) values in parts per thousand (%,
also termed ‘‘per mil’’) difference between sample and
standard ratios according to the formula
d13 Cðor d15 N or d34 SÞ ¼ ½ðRsample  Rstandard Þ=Rstandard 
 1000,
where R ¼ 13C/12C (or 15N/14N or 34S/32S). Thus samples
enriched in 13C (or 15N or 34S) are isotopically ‘‘heavy’’ and
have higher d, while samples depleted in 13C (or 15N or 34S)
are isotopically ‘‘light’’ and have lower d values (Hershey
and Peterson, 1996).
The adage ‘‘you are what you eat’’ is often used to
describe how stable isotopes are used in ecological studies.
The carbon, sulfur, and nitrogen values of consumers
within an aquatic ecosystem will be a direct reflection of the
isotope values at the bottom of the food chain, namely
primary producers (DeNiro and Epstein, 1978). Carbon
and sulfur isotope signatures for consumers will generally
be within72% of their diet, with the average being a slight
enrichment in the heavy isotope (i.e., a positive isotope
ratio shift of about 0.5%; Table 1). By comparison, the
nitrogen isotope ratio tends to be enriched by about 3.0%
from diet to consumer (Table 1).
Most terrestrial (C3) plants have a fairly consistent d13C
signature of approximately 28% based on utilization of
atmospheric CO2 (d13C of 8%) and subsequent isotope
fractionation during photosynthetic respiration (O’Leary,
1984, 1988). Therefore, biota in aquatic food webs that rely
on terrestrial carbon sources will have similar or slightly
enriched carbon isotope ratios. In contrast, aquatic
primary producers may exhibit a broad range of carbon
isotope values (e.g., approximately 1% to 28%) depend-
ing on the dissolved inorganic carbon source, with CO2
derived from limestone weathering near the positive
extreme and that derived from decomposition of terrestrial
detritus at the negative end of the range (Osmond et al.,
 ARTICLE IN PRESS
P.L. Orr et al. / Ecotoxicology and Environmental Safety 63 (2006) 175–188 177

Yukon Northwest Territories

FSP Fording Coal

Ala
ska
Alberta
FRO
Greenhills Mine

r
ive
din g R
FR
British Columbia

For
Elkford

Cr
Li ne
River Line Creek Mine
Elk River

Study Area
Fording

Montana
Idaho
LC
r.
C Washington
Lin e

Elk Valley

Elkview
GM Coal
M

x a n d e r Cr.

Legend
ic
he
lC

Sparwood
r.

Mine Waste
Ale

AC FSP Sampling Area

Miche

FSP Fording Settling Pond, lentic,

receives effluent from Fording Mine
l Cr.

Hosmer FRO Fording River Oxbow, lentic,

Leach Cr.

receives effluent from Fording Mine

FR Fording River, lotic, receives effluent

LCO from Fording Mine

Fernie Coal Mountain LC Line Creek, lotic, receives effluent

Mic h

from Line Creek Mine

Leac
el

Cr
GM Goddard Marsh, lentic, receives
h
Cr

BLW effluent from Elkview Coal

.

AC Alexander Creek, lotic, reference

R. area
er

ad
v
El k R i

he
at
Fl LCO Leach Creek Oxbow, lentic,
reference area

BLW Barnes Lake Wetland, lentic,

reference area
Flath

FHW Flathead Wetland, lentic, reference

e
ad R

area
FHW
.

0 5 10km
(approx. 20 kms)

Fig. 1. Study area in the Elk River watershed of British Columbia, Canada.

1981; Rounick and Winterbourn, 1986; Hershey and depending on factors such as CO2 availability (Guy et al.,
Peterson, 1996). Subsequent fractionation during photo- 1987; O’Leary, 1988), water flow (Osmond et al., 1981;
synthesis of aquatic plants also occurs to varying degrees, Raven et al., 1985), plant metabolic rate (MacLeod and
 ARTICLE IN PRESS
178 P.L. Orr et al. / Ecotoxicology and Environmental Safety 63 (2006) 175–188

Table 1
Differences (D) in tissue stable isotope values of consumers relative to diet reported in the scientific literature

Isotope Range Mean (%) Reference

Dd13Carbon 0.6% to +2.7% 0.8 DeNiro and Epstein, 1978

3% to+3% 0.2 Fry and Sherr, 1984; Peterson and Fry, 1987
2.7% to+3.4% 0.4 McCutchan et al., 2003
Not reported 0.2 (freshwater) France and Peters, 1997
Not reported 0.5 (estuary) France and Peters, 1997
Not reported 0.8 (coastal) France and Peters, 1997
Not reported 1.1 (open ocean) France and Peters, 1997
Dd34Sulfur 1% to +6% 0.2 Peterson and Fry, 1987
3.2% to 4% 0.5 McCutchan et al., 2003
Dd15Nitrogen 0.5% to +9.2% 3.0 DeNiro and Epstein, 1981
+1.3% to +5.3% 3.4 Minigawa and Wada, 1984
+3% to +5% 3.2 Peterson and Fry, 1987
Not reported 2–3 Fry, 1991
0.8% to 5.9% 2.0 McCutchan et al., 2003

Barton, 1998), and pH (Osmond et al., 1981), with lowest
carbon isotope values for algae being about 45% (Rau,
1980; Peterson and Fry, 1987). Similarly, sulfur and
nitrogen isotope ratios also vary among aquatic primary
producers, depending on the source of inorganic nitrogen
and sulfur, respectively. Relative reliance on allochthonous
(terrestrial) versus autochthonous (aquatic primary pro-
duction) nutrient sources will determine the stable isotope
values of food webs in different aquatic environments.
Therefore, stable isotope data were used to explain the
feeding relationships of biota within each habitat investi-
gated in the Elk River watershed and then related to the
observed Se concentrations to explain differences in Se
concentrations in mine effluent-exposed and reference lotic
and lentic habitats.
2. Material and methods
2.1. Field methods
Fish muscle tissue, invertebrate (whole body), sediment, and water
samples were collected from three reference areas between July 23 and
August 3, 2002 as part of an initial reconnaissance study of lentic areas of
the watershed (Flathead Wetland (FHW), Barnes Lake Wetland (BLW),
and Leach Creek Oxbow (LCO)) and areas receiving mine effluents
containing elevated Se concentrations (Fording River Oxbow (FRO),
Fording Settling Pond (FSP), and Goddard Marsh (GM)) (Fig. 1). A
second field program was undertaken July 22–28, 2003 to obtain (a)
additional samples of aquatic vegetation and invertebrates from the lentic
areas for Se analysis, (b) additional samples for stable isotope analysis
(SIA) from FSP and FRO to allow for temporal comparisons of 2002 and
2003 samples, and (c) samples from three lotic areas in the watershed,
namely Fording River (FR, mine-exposed), Line Creek (LC, mine-
exposed), and Alexander Creek (AC, reference).
Surface water samples were collected 30 cm below the water surface
directly into 250-mL polyethylene bottles for analysis of total Se. Surface
water samples taken for SIA of fine organic matter (FOM) were collected
as subsurface grabs directly into 500 mL polyethylene bottles and frozen.
Sediment samples were collected from lentic areas by compositing the
top 3 cm of three petite Ponar grabs (15.24 cm2 each grab) into 500-mL
polyethylene jars using a stainless-steel spoon. In lotic areas, sediment
samples were scooped directly into 500-mL polyethylene jars using a
stainless-steel spoon because sufficiently fine sediments for analysis were
available only in small patches between rocks. Sediment subsamples
(200 mg) for SIA were placed into polyethylene bags and frozen. Water
and sediment samples collected for Se analysis were stored in coolers or
refrigerators and then shipped in coolers to the laboratory.
Periphyton sometimes being composed of a thin layer of attached algae,
bacteria, and other microorganisms (epilithon) and other times being
composed of moss or macrophytic attached algae, depending on relative
local abundance, was collected from most habitats. This material was
scraped from one or more rocks to achieve minimum sample size
requirements and placed into polyethylene bags and frozen. Emergent
and/or submergent vascular plants (macrophytes) were also collected
manually if abundant.
In lentic areas, invertebrate samples for Se analysis and SIA were
collected using a petite Ponar grab (in deeper locations) and a 500 mm sieve
or by dip-netting (shallow areas). In lotic systems, invertebrates were hand
collected from under rocks in the stream. All collected invertebrates were
placed into polyethylene bags and kept inside a cooler until they could be
further sorted, in the field, into the dominant feeding habits of the
majority of species within each order: filterers, detritivores (including
shredders, gatherers, and scrapers), and predators according to the keys of
Merritt and Cummins (1996) and WLU (2000). Each sorted feeding guild
from each area was split into two prelabeled polyethylene bags and frozen.
Zooplankton samples were collected using a 65-mm mesh size plankton
net and stored frozen in 500-mL polyethylene containers.
Fish were collected using experimental gill nets (1.5- to 2.5-inch mesh,
lentic areas) or by fly rod angling (lotic areas). Total body length, fork
length, body weight, liver weight, gonad weight, and anomalies were
recorded at the field laboratory within hours of collection. Stomachs were
removed and preserved in 10% buffered formalin for subsequent stomach
content analysis. Ageing structures (otoliths and scales) were wrapped
separately in waxed paper and stored inside labeled envelopes for
subsequent age determination by North Shore Environmental Services
in Thunder Bay, Ontario, Canada.
2.2. Stable isotope analysis
Carbon, nitrogen, and sulfur stable isotope analyses were conducted at
the Environmental Isotope Laboratory at the University of Waterloo,
Waterloo, Ontario, Canada. Water samples were thawed and screened
through a 60-mm-mesh sieve to remove large zooplankton and debris, after
 ARTICLE IN PRESS
P.L. Orr et al. / Ecotoxicology and Environmental Safety 63 (2006) 175–188
which the water sample was freeze-dried. Sediment samples were thawed,
and major debris and invertebrate material were removed with the aid of
tweezers under a dissecting microscope. Plant, zooplankton, benthic
invertebrate, and fish samples were thawed and cleaned of visible debris
under a dissecting microscope. Water and sediment samples were acidified
with 10% hydrochloric acid to remove inorganic carbon. Of biological
samples, only benthic invertebrate samples that might contain a large
amount of inorganic carbon were acidified prior to analysis (e.g.,
gastropods and bivalves). All samples were then oven dried at a constant
temperature of 60 1C for 48 h and placed in a desiccator. Just prior to
analysis, the dried samples were pulverized using a Retsch MM2000 ball
mill grinder (F. Kurt Retsch GmbH & Co., Haan, Germany).
Approximately 1 mg of dried ground animal tissue was used in the
simultaneous analysis of carbon and nitrogen isotopes. Stable sulfur
isotopes required 3.5 mg of animal tissue due to the low sulfur levels in
animal tissue. The amount of dry weight material for sediment, FOM,
periphyton, and aquatic plant matter varied from 1 to 100 mg, depending
on total carbon, nitrogen, and sulfur content in each sample.
Dried pulverized samples were analyzed for nitrogen and carbon
isotopes on an Isochrom Continuous Flow Stable Isotope Mass Spectro-
meter (Micromass) coupled to a Carlo Erba Elemental Analyzer (CHNS-O
EA1108). Sulfur analyses were conducted using the Europa TracerMass/
Roboprep system. Dried, pulverized samples were combusted to CO2, N2,
or SO2 at 1000 1C in the elemental analyzer. The gases were separated
using different separation columns for separation of CO2 and N2
(Elemental Microanalysis Limited type E3003 for carbon, nitrogen,
hydrogen, and sulfur) versus SO2 (type E3002). The purified gases were
introduced into the isotope ratio mass spectrometer for analysis.
Sample isotope ratios were compared to those of standard gases
injected directly into the isotope ratio mass spectrometer before and after
the sample peaks, and d15N, d13C, and d34S values were calculated relative
to international standard materials obtained from the International
Atomic Energy Agency (IAEA), Austria. Ammonium sulfate standards
(IAEA-N1 and IAEA-N2) were used for d15N and were previously
calibrated against atmospheric nitrogen (d15NAIR; Mariotti, 1983).
Standard IAEA-CH6 (Anu Sugar Sucrose) was used for carbon and was
previously calibrated against carbonate rock from the Pee Dee Belemnite
(PDB) formation (d13CPDB; Craig, 1953). NBS-123 (Sphalerite ZnS) was
the sulfur standard and was previously calibrated against primordial
sulfur from the Canyon Diablo meteorite (d34SCDT; Rees et al., 1978).
Stable isotope ratios were expressed as delta values (d) with d15N, d13C,
and d34S values being relative to atmospheric 15N, PDB 13C, and Canyon
Diablo 34S, respectively.
2.3. Selenium analyses
Se analyses of water, sediment, and macrophyte samples were
completed by Aurora Laboratory Services of Vancouver, BC, Canada
using hydride vapor atomic absorption spectrophotometry (EPA Method
7000 series).
Periphyton, benthic invertebrates and fish tissue samples were analyzed
at the University of Missouri-Columbia Research Reactor Center. The
samples were received frozen and then freeze-dried, homogenized, and
analyzed by instrumental neutron activation analysis.
Se concentration results are presented in dry weight units unless
specified otherwise.
2.4. Estimation of trophic position and biomagnification potential
The consistency of nitrogen enrichment at each trophic transfer (Table
1) provides a convenient quantitative measure of relative trophic position
within a food web (Cabana and Rasmussen, 1994, 1996; Vander Zanden et
al., 1997; Vander Zanden and Rasmussen, 1999; Post, 2002). Nitrogen
isotope values alone cannot be used to compare the trophic position of
organisms in different aquatic environments because the d15N of primary
producers (organisms that convert inorganic N to organic N) are highly
variable among systems (Cabana and Rasmussen, 1996). This necessitates
179
that the isotopic signature of aquatic animals be measured relative to a
site-specific ‘‘baseline’’ d15N. Therefore, the trophic position of each
consumer within each area was estimated consistent with equations
presented by Vander Zanden et al. (1997), Vander Zanden and Rasmussen
(1999), and Post (2002),
Trophic position of consumer ¼ l þ ðd15 Nconsumer  d15 Nbase Þ=Dn,
where l is the trophic position of the organism used to estimate d15Nbase
(e.g., l ¼ 1 for primary producers), d15Nconsumer is measured directly,
and Dn is the enrichment in d15N per trophic level. For this study a trophic
enrichment value of 3.0% was adopted based on values presented in Table
1. The mean values for the available primary producers (periphyton and
macrophytes) were used to estimate the d15Nbase for each area. As the top
consumers collected in this project, the trophic position of fish in each area
also represented the total food chain length.
Trophic position can be correlated with contaminant concentrations to
trace the pathways of contaminant biomagnification to top predators in
aquatic food webs (Cabana and Rasmussen, 1994; Atwell et al., 1998;
Bowles et al., 2001; Power et al., 2002). Net bioaccumulation of an element
occurs when its trophic transfer is greater than that of carbon and the
concentration thus increases with increasing trophic level (Mason et al.,
2000). Biomagnification potential was estimated using the model (after
Broman et al., 1992; Rolff et al., 1993)
C biota ¼ AeBðtrophic positionÞ ,
where Cbiota is the Se concentration in the biota and A and B are
parameters estimated by linear regression after logarithmic transforma-
tion. The equations presented by Broman et al. (1992) and Rolff et al.
(1993) used measured d15N for the exponent, but estimated trophic
position (based on d15N) was substituted in this study to take into account
inter-area variability in baseline d15N. The model constant, A, is a scaling
factor that depends on the concentration of Se at the base of the food
chain (Rolff et al., 1993). The parameter B estimates biomagnification
potential, where B40 indicates that transfer of Se between trophic levels is
more efficient than biomass transfer (i.e., Se is biomagnified).
3. Results
3.1. Stable isotope data
The combined isotope data for all areas indicated more
enriched nitrogen isotope signatures among aquatic
animals in mine-exposed habitats than among those in
reference areas (Fig. 2). However, the mine-exposed areas
16
14
12
10
δ 15N (‰)
8
6
4
2
0
-40 -35 -30 -25 -20 -15
δ 13C (‰)
FSP FRO GM BLW FHW LCO FR LC AC
Fig. 2. d15N and d13C isotope values measured in fish, amphibian, and
benthic invertebrate tissue samples collected in all habitats sampled in the
Elk River watershed, British Columbia, in 2002 and 2003.
 ARTICLE IN PRESS
180 P.L. Orr et al. / Ecotoxicology and Environmental Safety 63 (2006) 175–188

were generally situated to the north of the watershed, while
most reference areas were located further south (Fig. 1).
Comparison of nitrogen isotope values against the distance
to FSP, the most northerly area sampled, suggested that
the pattern was simply a reflection of north–south
geographical position and associated subwatershed condi-
tions rather than effluent exposure (Fig. 3). The negative
relationship between distance to the south and d15N existed
even though the locations are not all located in the same
subwatershed (or even in the same watershed in the case of
FHW). No other separation of lentic or lotic areas or
reference or mine-exposed areas was evident in plots
showing carbon and sulfur or sulfur and nitrogen isotope
values of aquatic invertebrates and vertebrates (not
shown), suggesting that the differences in isotope signa-
16
14
12
10
δ N (‰)
tures among areas were indicative of factors other than
relative mine effluent exposure.
Mean carbon isotope values for aquatic primary
producers ranged from 19.6% to 38.3%, overlapping
with values typically associated with terrestrial plants
(approximately 28%), thus making it impossible to
distinguish between relative reliance on allochthonous
versus autochthonous carbon sources in many areas
(Table 2). However, shifts in carbon and sulfur isotope
values between benthic organisms (diet) and fish (con-
sumers) were generally consistent with ranges presented in
the literature (Tables 1 and 2). An exception was at GM
where a larger-than-expected trophic shift was indicated
based on average carbon and sulfur isotope values relative
to literature values (Table 2). This was likely related to
preferential feeding by resident sucker on specific inverte-
brates present in GM, some of which had more positive
signatures of both isotopes than was indicated by mean
values. Overall, the SIA data suggested that the fish
collected had fidelity to the habitats from which they were
obtained, which was an important consideration in the

8 y = -0.0575x + 10.102 investigation of the potential food web pathway(s) leading

15

R2 = 0.514, P<0.0005 to Se accumulation in fish in each habitat.

6
4
Food chain length was estimated in several ways for each
area based on average measured d15N values and the
2
assumption of an approximate d15N increase of 3.0 at each
0 trophic level (Table 3). The estimated number of trophic
0 20 40 60 80 100 120 140
Distance to FSP (km)
levels in each area was more variable when the calculation
included primary producers, which can exhibit wide
FSP FRO GM BLW FHW LCO FR LC AC
variations in isotope signatures within and among habitats
Fig. 3. Relationship between d15N of biological tissue samples and depending on the source and isotope signature of inorganic
relative distance from Fording Settling Pond (FSP), the most northerly carbon, nitrogen, and sulfur used by the plants, as
area sampled. discussed previously. Overall food web structure was

Table 2
Mean d13carbon and d34sulfur (%) of biological samples collected in lentic and lotic areas of the Elk River watershed, showing differences between the
stable isotope signatures of benthic invertebrates and fish among areas

Lentic Lotic

Exposed Reference Exposed Reference

FSP FRO GM BLW FHW LCO FR LC AC

Mean d13Carbon
Primary producers 23.8 19.6 27.4 32.4 32.5  20.6 32.5 38.3
Benthic invertebrates (all) 27.6 30.8 28.5 31.5 30.5 32.0 30.1 32.9 33.1
Amphibians (adult frogs only) — 29.2 26.7 — — 29.4 28.2 — —
Fish 26.6 28.9 23.7 28.5 29.1 30.4 30.0 30.3 30.4

Dd13Carbon: benthic invertebrates to fish 1.0 1.9 4.8a 3.0 1.4 1.6 0.1 2.6 2.7
34
Mean d Sulfur
Primary producers 6.5 0.96 7.2 4.7 6.3 — 3.1 1.3 4.7
Benthic invertebrates (all) 4.5 4.5 8.2 6.4 6.2 1.2 4.2 2.9 0.1
Amphibians (adult frogs only) — — — — — — 0.6 — —
Fish 3.2 3.5 5.9 2.7 5.0 2.9 2.0 2.9 1.3
Dd34Sulfur: Benthic Invertebrates to Fish 1.3 1.0 2.3a 3.7 1.2 1.7 2.2 0.0 1.2
a
Invertebrate species at Goddard Marsh exhibited a wide range of d13carbon and d34sulfur. The fish caught there may feed selectively on invertebrates
with less negative isotope signatures than is reflected by the mean value, resulting in trophic shifts more consistent with literature values (Table 1).
 ARTICLE IN PRESS
P.L. Orr et al. / Ecotoxicology and Environmental Safety 63 (2006) 175–188 181

Table 3
Estimates of food chain length in lentic and lotic habitats of the Elk River watershed based on d15nitrogen measured at different trophic levels

Average d15Nitrogen Lentic Lotic

Exposed Reference Exposed Reference

FSP FRO GM BLW FHW LCO FR LC AC

Primary producers 6.1 7.2 6.5 0.02 0.1 — 4.0 2.3 0.15
Benthic invertebrates (primary consumers only) 7.8 10.3 8.7 2.8 1.1 3.6 7.4 4.6 4.6
Ephemeroptera+Chironomidae 6.7 10.4 9.2 3.5 0.8 3.6 8.0 4.1 3.7
Benthic invertebrates (all) 7.8 10.7 9.3 3.5 2.3 4.5 7.7 5.3 4.6
Fish 10.4 13.2 12.0 7.5 4.7 8.8 9.9 9.4 8.6
Dd15Nitrogen: primary producers to benthos (primary consumers) 1.7 3.1 2.2 2.8 1.0 — 3.4 2.3 4.8
Dd15Nitrogen: primary producers to fish 4.3 6.0 5.5 7.5 4.6 — 5.9 7.1 8.8
Estimated trophic position of fish (No. of trophic levels)a 2.4 3.0 2.8 3.5 2.5 — 3.0 3.4 3.9
Dd15Nitrogen: benthic invertebrates (all) to fish 2.6 2.5 2.7 4.0 2.4 4.3 2.2 4.1 4.0
Estimated trophic position of fish (No. of trophic levels)b 2.9 2.8 2.9 3.3 2.8 3.4 2.7 3.4 3.3

Dd15Nitrogen: ‘Ephemeroptera+Chironomidae’ to fish 3.7 2.8 2.8 4.0 4.0 5.3 1.9 5.4 4.9
Estimated trophic position of fish (No. of trophic levels)c 3.2 2.9 2.9 3.3 3.3 3.8 2.6 3.8 3.6
a
Estimated food chain length between fish and primary producers. Calculated as Dd15nitrogen/3+1, assuming an approximate increase in Dd15nitrogen
of 3 at each increasing trophic level as shown in Table 1.
b
Estimated food chain length between fish and benthic invertebrates. Calculated as Dd15nitrogen/3+2, assuming an approximate increase in
15
Dd nitrogen of 3 at each increasing trophic level as shown in Table 1.
c
Estimated food chain length between fish and ‘Ephemeroptera+Chironomidae’. Calculated as Dd15nitrogen/3+2, assuming an approximate increase
in Dd15nitrogen of 3 at each increasing trophic level as shown in Table 1.

similar in lotic and lentic habitats, as indicated by the
presence of primary and secondary consumer benthic
species and one dominant fish species (cutthroat trout at
all areas but FHW and GM, where longnose sucker was
obtained, and LCO, where brook trout was obtained), all
of which appeared to feed predominantly on resident
benthic invertebrates. This was supported by stomach
content analysis (not presented). Estimates of food chain
length generally indicated three trophic levels.
3.2. Selenium concentrations in the food web
The water concentrations of Se measured in 2002 and
2003 at lentic exposure and lotic reference and exposure
areas were within the range of concentrations reported in
previous monitoring at those locations (Table 4). Reference
lentic areas were not monitored previously, but values
measured in 2002 and 2003 were similar to those measured
at the lotic reference area (AC). Elk River watershed
reference area concentrations were slightly greater than
those reported for reference areas in other studies
(0.1–0.4 mg/L; USDOI, 1998), likely due to the naturally
seleniferous geology of the area. In general, aqueous Se
concentrations were greater in exposure versus reference
areas, although Se concentrations in the exposure areas
were highly variable, reflecting the relative proximity to
mine site discharges.
As with water, sediment Se concentrations in exposure
areas were variable and ranged from levels comparable to
reference (2–3 mg/kg at FSP) to a maximum of 26 mg/kg at
GM (Table 4).
The animals in lentic exposure areas had, on average,
much higher tissue Se concentrations than those in lentic
reference areas (Fig. 4, Table 5). In contrast, the biota in
lotic exposure areas had tissue levels of Se similar to those
in both the lotic reference area and the lentic reference area
(Fig. 4), indicating that Se is taken up more readily into the
food web in lentic than in lotic environments. However,
primary producers in lentic exposure areas did not contain
higher Se concentrations than those in the other environ-
ments (Table 6), indicating that the key process of Se
incorporation into the food web occurs through a pathway
other than direct consumption of primary producers.
The Se concentrations of aquatic plants and animals in
lentic and lotic habitats were compared to relative trophic
position of each organism based on d15N. The biota at
higher trophic positions in lentic exposure areas accumu-
lated much greater Se concentrations that those in other
habitats (Fig. 5).
Aqueous exposure concentrations were generally lower
in the lotic than in some lentic exposure areas, so the data
were evaluated separately for one lotic and one lentic area
with fairly similar concentrations of Se in water. Higher Se
concentrations were evident among biota in the FRO
(lentic area) than in LC (lotic area) even though the latter
had marginally higher aqueous Se levels (Fig. 6, Table 4).
Average values computed for organisms classified as
benthic detritivores, benthic predators, and fish did not
suggest that biomagnification occurs among Elk River
 ARTICLE IN PRESS
182 P.L. Orr et al. / Ecotoxicology and Environmental Safety 63 (2006) 175–188

Table 4
Water and sediment quality data for present study (2002–2003) compared to aqueous selenium concentrations reported by EVS (2002) for same locations.

Habitat Area type Specific location Location Total aqueous selenium (mg/L) Total sediment selenium
type identifier (mg/kg dry weight)
1995–2001 (EVS 2002) This Study

Number of Mean (range) 2002 (n ¼ 1 2003 (n ¼ 1 2002 (n ¼ 1 2003 (n ¼ 1

Samples for each area) for each area) for each area) for each area)

Lentic Exposure Fording River FRO 5 9.6 (4–14) 9.9 4.4 7.0 8.8
Oxbow
Fording Settling FSP 17 25.4 (o1–54) 50.0 42.0 3.0 2.6
Pond (Clode
Pond)
Goddard Marsh GM 16 63.5 (7–106) 82.0 94.0 26.0 —

Reference Flathead Pond FHW — — 0.7 0.7 3.0 —

Barns Lake BLW — — 0.5 0.5 2.0 —
Wetland
Leach Creek LCO — — 1.4 0.9 3.0 —
Oxbow

Lotic Exposure Line Creek LC 43 15.1 (4.6–25) — 20.9 — 2.1

Fording River FR 7 14.5 (7–19) — 20.1 — 2.1

Reference Alexander Creek AC 4 1.4 (0.5–3.7) — 0.9 — 0.9

80 Lentic Reference (BLW,FHW,LCO) and lotic exposure habitats (Rolff et al., 1993), but to a
Lentic Exposure (FSP,FRO,GM) greater extent in lentic habitats.
Lotic Reference (FR,LC)
Lotic Exposure (AC) It is noteworthy that the filtering benthic invertebrates
70
(Pisidium) obtained at exposure area FSP (7.2 mg/kg) and
reference area LCO (1.8 mg/kg) had lower Se concentra-
60 tions than other benthic invertebrates at those locations (91
and 14 mg/kg, respectively). These bivalves derive their
50 food from particulate matter transported in the water
column, whereas the other invertebrates sampled rely on
Se (ppm)

food sources that are more closely associated with the

40
sediment–detrital food web. It is also noteworthy that the
isotope signatures of bulk sediment and cutthroat trout
30 were more similar in the three lentic areas where this
species was dominant (Dd13C of only 1.3–3.3% between
20 sediment and fish) compared to the three lotic areas (Dd13C
of 4.9–5.8%), also suggesting a closer dietary link to
sediments in lentic areas.
10

0 4. Discussion
Water Sediment Benthic Benthic Benthic Benthic Amphibians Cutthroat
Plants Filterers Detritivores Predators Trout

Fig. 4. Mean selenium concentrations in water, sediment, algae, benthic
invertebrates, amphibians, and fish from the lentic and lotic reference and
exposure areas of the Elk River watershed.
watershed biota (Fig. 4, Table 5). Furthermore, the greatest
tissue Se concentrations were sometimes observed among
specific benthic invertebrates (e.g., Ephemeroptera with
165.4 mg/kg at GM). However, the positive b values in the
exponential equations shown in Figs. 5 and 6 (y ¼ aebx )
indicated that Se biomagnification occurred in both lentic
SIA identified that the food chain structure in both lotic
and lentic habitats was relatively simple, involving three
trophic levels: (1) allochthonous and/or autochthonous
primary production, (2) primary consumers (filtering,
grazing, and detritivorous benthic invertebrates), and (3)
secondary consumers (predator invertebrates, amphibians,
and fish). SIA also indicated that the fish collected in this
study likely derive much of their nutrition from the lentic
and lotic habitats from which they were collected, allowing
for characterization of potential differences in food chain
dynamics giving rise to elevated concentrations of Se in fish
tissues in lentic but not lotic exposure areas.
 ARTICLE IN PRESS
P.L. Orr et al. / Ecotoxicology and Environmental Safety 63 (2006) 175–188 183

Table 5
Mean tissue selenium concentrations observed in biota representing different trophic levels (mg/kg dry weight, sample sizes in parentheses)

Lentic Lotic

Exposed Reference Exposed Reference

FSP FRO GM BLW FHW FR LC AC

Benthic detritivores 91.3 (2) 26.6 (3) 96.4 (2) 8.5 (2) 3.7 (1) 7.1 (1) 7.3 (1) 9.6 (1)
Benthic predators — — 42.6 (1) 8.2 (1) 6.6 (2) 6.9 (1) 2.7 (1) 4.5 (1)
Fish (muscle) 41.6 (6) 25.7 (6) 33.6 (3) 4.6 (3) 3.9 (3) 8.2 (3) 7 (3) 4.9 (3)

Table 6
Selenium concentrations measured in samples of aquatic primary producers (mg/kg dry weight)

Lentic Lotic

Exposed Reference Exposed Reference

FSP FRO GM BLW FHW LCO FR LC AC

Algae
Epilithon 4.47 7.63 — — — — 5.43 2.19 —
Chara 6.5 — — — — —
Cladophora — 3.47 3.21 4.40 — — 1.11 — 4.49

Moss — — — — 3.89 3.67 7.94 — —

Macrophytes
Carex 3.9 — 3.9 — — — — — —
Equisetum 3.9 — 4.3 — — — — — —
Typha — — 12.3 — — — — — —

All Sites 80
120
Tissue Se (mg/kg dry weight)

70
100
60
80

50 FRO ( )
Se (mg/kg)

60
y = 4.1216e0.7986x
2
40 R = 0.5178
40

30
20
LC ( )
20 y = 1.1169e0.7362x
0
R2 = 0.6009
0.0 0.5 1.0 1.5 2.0 2.5 3.0 3.5 4.0 4.5
Trophic Position 10

Lentic Reference Lentic Exposure Lotic Reference Lotic Exposure

0
0.0 0.5 1.0 1.5 2.0 2.5 3.0 3.5
Fig. 5. Selenium concentrations measured in biological tissue samples Trophic Position
collected from lentic and lotic habitats of the Elk River watershed, relative FRO LC FRO Exponential Trendline LC Exponential Trendline
to trophic position. Trophic position was calculated from d15nitrogen
values of each sample. Fig. 6. Trophic position and selenium concentration of each biological
tissue sample collected at FRO (exposed lentic area) versus LC (exposed
lotic area).
The food chain lengths of lentic areas were similar to or
shorter than those of lotic areas, so the hypothesis of
elevated Se levels potentially being associated with greater exposure areas, when measured d15N (indicating actual
food chain length in lentic areas was not supported by the trophic position), rather than assumed dietary relation-
data. Se biomagnification was shown, particularly in lentic ships, was considered. These results indicated that general
 ARTICLE IN PRESS
184 P.L. Orr et al. / Ecotoxicology and Environmental Safety 63 (2006) 175–188
assumptions about the feeding behavior (e.g., detritivore
versus predator) of benthic invertebrate orders containing
species with a wide variety of feeding characteristics cannot
be made. Stable nitrogen isotope data provided a
more reliable means of evaluating trophic feeding relation-
ships by indicating actual rather than assumed dietary
relationships.
Greater macrophyte abundance in lentic areas was
consistent with the tendency of watershed wetlands to act
as reservoirs for a large proportion of watershed nutrients.
The lentic exposure environments were associated with
elevated concentrations of Se in benthic invertebrates but
not in primary producers, indicating that the key process of
Se incorporation into the food web occurs through a
pathway other than direct consumption of primary
producers. Instead, this study provides evidence of the
importance of sediment–detrital processes, which give rise
to elevated concentrations in benthos, likely via uptake of
microorganisms containing elevated Se concentrations
and/or via direct uptake by benthos of Se from water
(porewater or water near the sediment interface) or
incidental sediment ingestion. This conclusion was sup-
ported by the findings of lower Se accumulation in filter-
feeding bivalves than in benthic detritivores, enhanced
biomagnification of Se in lentic compared to lotic exposure
areas, and greater similarity between the isotope signatures
of bulk sediment and that of cutthroat trout in lentic
compared to lotic areas.
Taken together, the data related to Se speciation and
relative Se uptake by biota presented in the literature are
consistent with the conclusions of this study. Uptake of the
dissolved oxyanions (selenate and selenite) by algae,
macrophytes and bacteria and subsequent transformation
to selenoamino acids (e.g., selenocysteine and seleno-
methionine) are cited as key pathways for biotransforma-
tion of dissolved inorganic Se into organoselenium. This is
because selenium is readily taken up from water by algae
(Sandholm et al., 1973; Nassos et al., 1980; Foe and
Knight, 1986; Riedel et al., 1991; Besser et al., 1993; Herbel
et al., 2002), aquatic plants (Allen, 1991; Ornes et al., 1991;
Herbel et al., 2002), and bacteria (Bender et al., 1991)
under laboratory and field conditions. Se speciation affects
the rate at which it is assimilated by plants, with uptake
rate increasing from least to most rapid in the order of
selenate versus selenite versus organoselenide (Ogle et al.,
1988; Lemly et al., 1993; Maier et al., 1993; Alaimo et al.,
1994; Bowie et al., 1996; Kiffney and Knight, 1990; Riedel
et al., 1996). Se taken up in the form of oxyanions by plants
or microorganisms is then largely transformed into
selenoamino acids and subsequently incorporated into
proteins (Butler and Peterson, 1967; Wrench, 1978; Bottino
et al., 1984). Thus, it has been previously concluded,
although not specifically demonstrated, that bacteria and
aquatic plants are primarily responsible for introducing
organoselenides into aquatic food webs and account for
much of the biogeochemical cycling of Se that occurs in
natural systems (Bowie et al., 1996).
Consumer aquatic organisms (e.g., invertebrates and
fish) can also take up Se directly from water, and, as with
microbes and plants, selenate is the least bioavailable form
and organoselenium is the most bioavailable form (Niimi
and LaHam, 1976; Ingersoll et al., 1990; Luoma et al.,
1992; Besser et al., 1993; Lemly et al., 1993; Maier et al.,
1993). Therefore, some of the Se taken up by aquatic biota
may occur via waterborne organoselenium compounds
released from living algae or by the necrosis of cells
(Cutter, 1992; Besser et al., 1994). Furthermore, environ-
ments in which a substantial proportion of Se is present as
aqueous organoselenides will lead to greater accumulation
of Se from water by both primary producers and
consumers.
In general, however, the dominant route of Se uptake by
aquatic consumer organisms is from food (Sandholm et al.,
1973; Besser et al., 1993; Bowie et al., 1996), in which Se
also tends to be largely in an organoselenide form (Fan et
al., 2002). Se in suspended and bottom particles is a
critically important phase because it represents the
food base for the smallest consumers (zooplankton and
benthic invertebrates, respectively) in aquatic food webs
(Lemly, 1985; Luoma et al., 1992; Fan et al., 2002),
which are in turn consumed by organisms at higher trophic
levels.
Bottom sediments are the dominant sink for Se in
aquatic ecosystems via association with suspended parti-
culate matter and subsequent deposition (Besser et al.,
1989; Canton and Van Derveer, 1997). Se tends to sorb
most rapidly to fine- rather than coarse-textured
substrate (Besser et al., 1989), consistent with greater
surface area for Se sorption and the elevated Se concentra-
tions in some lentic area sediments in the Elk River
watershed. Bottom sediments are considered an important
link and exposure source to the benthic-driven food
web (Saiki et al., 1993; Lemly, 1993, 1996; Alaimo et al.,
1994; Maier and Knight, 1994; Bowie et al., 1996;
Sappington, 2002; Hamilton et al., 2004). Detritus can
contain high Se concentrations in Se-contaminated
environments (e.g., up to 440 mg/kg reported by Saiki
(1986) and Saiki and Lowe (1987)) and benthic inverte-
brates readily accumulate Se from detritus (this study;
Alaimo et al., 1994)
The chemical form of Se in sediments is controlled by the
nature of the ambient microbial community (i.e., either
selenooxyanion reducers or elemental Se oxidizers), which
in turn is dictated by redox conditions (Schlekat et al.,
2000; May et al., 2001). Some studies have indicated that
dissimilatory reduction of Se by bacteria may be important
in transforming selenooxyanions in overlying water or
porewater to sediment-associated elemental Se (Maiers
et al., 1988; Oremland et al., 1990; Fan et al., 2002; Amweg
et al., 2003) and then to organoselenide (Foda et al., 1983;
Schlekat et al., 2000; Fan et al., 2002) in reducing
environments.
On the other hand, analysis of Se stable isotope ratios in
a California estuary showed that reduction of soluble Se
 ARTICLE IN PRESS
P.L. Orr et al. / Ecotoxicology and Environmental Safety 63 (2006) 175–188
from overlying waters is not always the dominant process
by which Se is incorporated into sediments (Johnson et al.,
2000). Instead, it appears that Se assimilation by plants and
algae, followed by deposition and mineralization, may be
the dominant transformation pathway responsible for
accumulation of reduced forms of Se in the sediments of
some wetlands (Herbel et al., 2002). This is supported by
other studies in which it was concluded that accumulation
of Se in the top layer of sediment is generally the result of
deposition of dead organic material from the water column
and incorporation in the detrital food chain (Kiffney and
Knight, 1990; Oremland et al., 1990; Bender et al., 1991;
Graham et al., 1992; Stephens, 1996; Zawislanski et al.,
2001). Dead periphyton can contribute to the detrital food
web (Allan, 1995) as can terrestrial plant litter (Allan, 1995;
Waters, 1995). Very little aquatic macrophyte biomass is
grazed directly by herbivores; rather, the plants die and
become important contributors to the detrital food chain
(Teal, 1962; Hamilton and Buhl, 2003). Bacteria, fungi, and
other microbes then colonize the detrital material as part of
benthic nutrient cycling in aquatic systems (Kaushik and
Hynes, 1971; Baker and Bradnum, 1976; Bender et al.,
1991). Organisms higher in the food web, including
detritivores such as midge larvae, are thought to utilize
mainly the microbial biomass component of detritus
(Odum and de al Cruz, 1967; Baker and Bradnum, 1976;
Fenchel and Jorgensen, 1977; Harper et al., 1981;
Foda et al., 1983). Thus, some authors have concluded
that uptake of Se by benthic organisms from the
detrital food chain is one of the most important pathways
by which Se is recycled in aquatic environments (Canton
and Van Derveer, 1997; Hamilton and Lemly, 1999)
playing an extremely important role in the eventual
bioaccumulation and toxicity of Se in fish and birds
(Lemly, 1993; Saiki et al., 1993; Alaimo et al., 1994;
Maier and Knight, 1994; Lemly, 1996). Woock (1984)
found that caged golden shiner (Notemigonus crysoleucas)
exposed to sediment accumulated more Se than those
suspended in cages above the sediment, demonstrating the
importance of the sediment-linked food web in Se
accumulation.
Therefore, it is concluded that higher Se uptake in lentic
exposure areas of the Elk Valley occurs because their
hydrological characteristics enhance biotransformation of
Se oxyanions to organoselenium via macrophytes and
microbiota associated with substrate surfaces. In addition,
the relatively slow hydraulic retention time of lentic
compared to lotic areas promotes uptake and recycling
via sediment–detrital pathways. Consistent with Hamilton
(2002), this study also suggests that elevated concentrations
discharged to lotic habitats may pose greater risk to biota
in off stream and downstream lentic habitats of some
watersheds than in lotic reaches of the same watershed. The
overall risk to watershed biota will depend on the mix of
aquatic habitat types present, their hydrological connec-
tions, and the relative utilization by biota of each type of
habitat.
185
Acknowledgments
This study was partially funded by an Industrial
Research Fellowship awarded to Karin Guiguer by the
National Science and Engineering Research Council. The
authors thank the Elk Valley Coal Corp. and the other
members of the Elk Valley Selenium Task Force for their
technical input and support during this project. We also
thank the numerous other individuals who provided input
to the project and/or reviewed drafts of the full technical
report, notably D. Lemly (US Forest Service), W. Adams
(Rio Tinto), A. Fairbrother (US EPA), S. Chandra
(U. Nevada-Reno), K. Munkittrick (U. New Brunswick),
G. Dixon (U. Waterloo), A. Farwell (U. Waterloo),
M. Power (U. Waterloo), and D. Laine (U. Ottawa).
References
Alaimo, J., Ogle, R.S., Knight, A.W., 1994. Selenium uptake by larval
Chironomus decorus from a Ruppia maritima-based benthic detrital
substrate. Arch. Environ. Contam. Toxicol. 27, 441–448.
Allan, J.D., 1995. Stream Ecology–Structure and Function of Running
Waters. Chapman & Hall, New York 388pp.
Allen, K.N., 1991. Seasonal variation of selenium in outdoor experimental
stream–wetland systems. J. Environ. Qual. 20, 865–868.
Amweg, E.L., Stuart, D.L., Weston, D.P., 2003. Comparative bioavail-
ability of selenium to aquatic organisms after biological treatment of
agricultural drainage water. Aquat. Toxicol. 63, 13–25.
Arthur, J.R., Beckett, G.J., 1994. New metabolic roles for selenium. Proc.
Nutr. Soc. 53, 615–624.
Atwell, L., Hobson, K.A., Welch, H.E., 1998. Biomagnification and
bioaccumulation of mercury in an arctic marine food web: insights
from stable nitrogen isotope analysis. Can. J. Fish. Aquat. Sci. 55,
1114–1121.
Baker, J.H., Bradnum, L.A., 1976. The role of bacteria in the nutrition of
aquatic detritivores. Oecologia 24, 95–104.
Bender, J., Gould, J.P., Vatcharapijarn, Y., Saha, G., 1991. Uptake
transformation and fixation of Se(VI) by a mixed selenium-tolerant
ecosystem. Water Air Soil Pollut. 59, 359–367.
Besser, J.M., Huckins, J.N., Little, E.E., LaPoint, T.W., 1989. Distribu-
tion and bioaccumulation of selenium in aquatic microcosms. Environ.
Pollut. 62, 1–12.
Besser, J.M., Canfield, T.J., La Point, T.W., 1993. Bioaccumulation of
organic and inorganic selenium in a laboratory food chain. Environ.
Toxicol. Chem. 12, 57–72.
Besser, J.M., Huckins, J.N., Clark, R.C., 1994. Separation of selenium
species released from Se-exposed algae. Chemosphere 29, 771–780.
Bottino, N.R., Banks, C.H., Irgolic, K.J., Micks, P., Wheeler, A.E.,
Zingaro, R.A., 1984. Selenium-containing amino-acids and proteins in
marine-algae. Phytochemistry 23, 2445–2452.
Bowie, G.L., Sanders, J.G., Riedel, G.F., Gilmour, C.C., Breitburg, D.L.,
Cutter, G.A., Porcella, D.B., 1996. Assessing selenium cycling and
accumulation in aquatic systems. Water Air Soil Pollut. 90, 93–104.
Bowles, K.C., Apte, S.C., Maher, W.A., Kawei, M., Smith, R., 2001.
Bioaccumulation and biomagnification of mercury in Lake Murray,
Papua New Guinea. Can. J. Fish. Aquat. Sci. 58, 888–897.
Broman, D., Näf, C., Rolff, C., Zebühr, Y., Fry, B., Hobbie, J., 1992.
Using ratios of stable nitrogen isotopes to estimate bioaccumulation
and flux of polychlorinated dibenzo-p-dioxins (PCDDs) and dibenzoi-
furans (PCDFs) in two food chains from the northern Baltic. Environ.
Toxicol. Chem. 11, 331–345.
Butler, G.W., Peterson, P.J., 1967. Uptake and metabolism of inorganic
forms of selenium-75 by Spirodela oligorrihza. Aust. J. Biol. Sci. 20,
7786.
 ARTICLE IN PRESS
186 P.L. Orr et al. / Ecotoxicology and Environmental Safety 63 (2006) 175–188
Cabana, G., Rasmussen, J.B., 1994. Modelling food chain structure and
contaminant bioaccumulation using stable nitrogen isotopes. Nature
372, 255–257.
Cabana, G., Rasmussen, J.B., 1996. Comparison of aquatic food chains
using nitrogen isotopes. Proc. Natl. Acad. Sci. USA 93, 10844–10847.
Canton, S.P., Van Derveer, W.D., 1997. Selenium toxicity to aquatic life:
an argument for sediment-based water quality criteria. Environ.
Toxicol. Chem. 16, 1255–1259.
Craig, H., 1953. The geochemistry of the stable carbon isotopes. Geochim.
Cosmochim. Acta 3, 53–92.
Cutter, G.A., 1992. Kinetic controls on metalloid speciation in seawater.
Mar. Chem. 40, 65–80.
Demayo, A., Taylor, M.C., Reeder, S.W., 1979. Inorganic chemical
substances: selenium. In: Guidelines for Surface Water Quality.
Environment Canada, Inland Waters Directorate, Water Quality
Branch, Ottawa.
DeNiro, M.J., Epstein, S., 1978. Influence of diet on the distribution of
carbon isotopes in animals. Geochim. Cosmochim. Acta 42, 495–506.
DeNiro, M.J., Epstein, S., 1981. Influence of diet on the distribution of
nitrogen isotopes in animals. Geochim. Cosmochim. Acta 45, 341–351.
de Souza, M.P., Chu, D., Zhao, M., Zayed, A.M., Ruzin, S.E., Schichnes,
D., Terry, N., 1999. Rhizosphere bacteria enhance selenium accumula-
tion and volatilization by Indian mustard. Plant Physiol. 119, 565–573.
EVS (EVS Environmental Consultants), 2002a. Elk Valley selenium
monitoring study: draft. Prepared for the Elk Valley Mines Environ-
mental Management Committee, April 2002.
EVS (EVS Environmental Consultants), 2002b. Elk Valley mines selenium
trend analysis. Prepared for the Elk Valley Mines Environmental
Management Committee, June 2002.
Fan, T.W.-M., The, S.J., Hinton, D.E., Higashi, R.M., 2002. Selenium
biotransformations into proteinaceous forms by foodweb organisms of
selenium-laden drainage waters in California. Aquat. Toxicol. 57,
65–84.
Fenchel, T.M., Jorgensen, B.B., 1977. Detritus food chains of aquatic
ecosystems: the role of bacteria. In: Alexander, M. (Ed.), Advances in
Microbial Ecology. Plenum Press Publishing Corp., New York, pp.
10–58.
Foda, A., Vandermeulen, J., Wrench, J.J., 1983. Uptake and conversion of
selenium by a marine bacterium. Can. J. Fish. Aquat. Sci. 40 (Suppl.
2), 215–220.
Foe, C., Knight, A.W., 1986. Selenium bioaccumulation, regulation, and
toxicity in the green alga, Selenastrum capricornutum, and dietary
toxicity of the contaminated alga to Daphnia magna. In: Selenium in
the Environment, Publication No. CAT1/860201. California Agricul-
tural Technology Institute, California State University, Fresno, CA,
pp. 77–88.
France, R.L., Peters, R.H., 1997. Ecosystem differences in the trophic
enrichment of 13C in aquatic food webs. Can. J. Fish. Aquat. Sci. 54,
1255–1258.
Fry, B., 1991. Stable isotope diagrams of freshwater food webs. Ecology
72, 2293–2297.
Fry, B., Sherr, E.B., 1984. d13C measurements as indicators of carbon
flow in marine and freshwater ecosystems. Contrib. Mar. Sci. 27,
13–47.
Graham, R.V., Blaylock, B.G., Hoffman, F.O., Frank, M.L., 1992.
Comparison of selenomethionine and selenite cycling in freshwater
experimental ponds. Water Air Soil Pollut. 62, 25–42.
Guy, R.D., Fogel, M.F., Berry, J.A., Hoering, T.C., 1987. Isotope
fractionation during oxygen production and consumption by plants.
Prog. Photosyn. Res. 3, 597–600.
Hamilton, S.J., 2002. Rationale for a tissue-based selenium criterion for
aquatic life. Aquat. Toxicol. 57, 85–100.
Hamilton, S.J., Buhl, K.J., 2003. Selenium and other trace elements in
water, sediment, aquatic plants, aquatic invertebrates, and fish from
streams in southeastern Idaho near phosphate mining operations: May
2001. Final Report as part of the USGS Western US Phosphate
Project, May 23, 2003. US Geological Survey, US Department of the
Interior.
Hamilton, S.J., Lemly, A.D., 1999. Water-sediment controversy in setting
environmental standards for selenium. Ecotoxicol. Environ. Saf. 44,
227–235.
Hamilton, S.J., Holley, K.M., Buhl, K.J., Bullard, F.A., Weston, L.K.,
McDonald, S.F., 2004. Evaluation of flushing of a high-selenium
backwater channel in the Colorado River. Environ. Toxicol. 19, 51–81.
Harper, R.M., Fry, J.C., Lerner, M.A., 1981. A bacteriological investiga-
tion to elucidate the feeding biology of Nais variabilis (Oligochaeta:
Naididae). Freshwater Biol. 11, 227–236.
Herbel, M.J., Johnson, T.M., Tanji, K.K., Gao, S., Bullen, T.D., 2002.
Selenium stable isotope ratios in California agricultural drainage water
management systems. J. Environ. Qual. 31, 1146–1156.
Hershey, A.E., Peterson, B.J., 1996. Stream food webs. In: Hauer, F.R.,
Lamberti, G.A. (Eds.), Stream Ecology. Academic Press, San Diego,
CA, pp. 511–530.
Ingersoll, C.G., Dwyer, F.J., May, T.W., 1990. Toxicity of inorganic and
organic selenium to Daphnia magna and Chironomus riparius (Diptera).
Environ. Toxicol. Chem. 9, 1171–1181.
Johnson, T.M., Bullen, T.D., Zawislanski, P.T., 2000. Selenium stable
isotope ratios as indicators of sources and cycling of selenium: results
from the Northern Reach of San Francisco Bay. Environ. Sci.
Technol. 34, 2075–2079.
Kaushik, N.K., Hynes, H.B., 1971. The fate of dead leaves that fall into
streams. Arch. Hydrobiol. 68, 465–515.
Kennedy, C.J., McDonald, L.E., Loveridge, R., Strosher, M.M., 2000.
The effect of bioaccumulated selenium on mortalities and deformities
in the eggs, larvae and fry of a wild population of cutthroat trout
(Oncorhynchus clarki lewisi). Arch. Environ. Contam. Toxicol. 39,
46–52.
Kidd, K.A., Schindler, D.W., Muir, D.C.G., Lockhart, W.L., Hesslein,
R.H., 1995. High concentrations of toxaphene in fishes from a
subarctic lake. Science 269, 240–242.
Kiffney, P., Knight, A., 1990. The toxicity and bioaccumulation of
selenate, selenite and seleno-L-methionine in the cyanobacterium
Anabaena flosaquae. Arch. Environ. Contam. Toxicol. 19, 488–494.
Lemly, A.D., 1985. Toxicology of selenium in a freshwater reservoir:
implications for environmental hazard evaluations and safety.
Ecotoxicol. Environ. Saf. 34, 223–227.
Lemly, A.D., 1993. Guidelines for evaluating selenium data from aquatic
monitoring and assessment studies. Environ. Monit. Assess. 28,
83–100.
Lemly, A.D., 1996. Selenium in aquatic organisms. In: Beyer, W.N.,
Heinz, G.H., Redmon-Norwood, A.W. (Eds.), Environmental Con-
taminants in Wildlife—Interpreting Tissue Concentrations. CRC
Lewis Publishers, Boca Raton, FL, pp. 427–445.
Lemly, A.D., 1999. Selenium transport and bioaccumulation in aquatic
ecosystems: a proposal for water quality criteria based on hydrological
units. Ecotoxicol. Environ. Saf. 42, 150–156.
Lemly, A.D., Smith, G.J., 1987. Aquatic cycling of selenium: implications
for fish and wildlife, fish and wildlife leaflet 12. US Fish and Wildlife
Service, Washington, DC.
Lemly, A.D., Finger, S.E., Nelson, M.K., 1993. Sources and impacts of
irrigation drainwater contaminants in arid wetlands. Environ. Toxicol.
Chem. 12, 2265–2279.
Lillebo, H.P., Shaner, S., Carlson, D., Richard, N., DuBowy, P., 1988.
Regulation of agricultural drainage to the San Joaquin River.
State Water Resources Control Board, SWRCB Order No.
W.Q. 85–1.
Luoma, S.N., Johns, C., Fisher, N., Steinberg, N.A., Oremland, R.S.,
Reinfelder, J.R., 1992. Determination of selenium bioavailability to a
benthic bivalve from particulate and solute pathways. Environ. Sci.
Technol. 26, 485–491.
MacLeod, N.A., Barton, D.R., 1998. Effects of light intensity, water
velocity, and species composition on carbon and nitrogen
stable isotope ratios in periphyton. Can. J. Fish Aquat. Sci. 55,
1919–1925.
Maier, K.J., Knight, A.W., 1994. Ecotoxicology of selenium in freshwater
systems. Rev. Environ. Contam. Toxicol. 134, 31–48.
 ARTICLE IN PRESS
P.L. Orr et al. / Ecotoxicology and Environmental Safety 63 (2006) 175–188
Maier, K.J., Foe, C.G., Knight, A.W., 1993. Comparative toxicity of
selenate, selenite, seleno-DL-methionine and seleno-DL-cysteine to
Daphnia magna. Environ. Toxicol. Chem. 12, 755–763.
Maiers, D.T., Wichlacz, P.L., Thompson, D.L., Bruhn, D.F., 1988.
Selenate reduction by bacteria from a selenium-rich environment.
Appl. Environ. Microbiol. 54, 2591–2593.
Mariotti, A., 1983. Atmospheric nitrogen is a reliable standard for natural
15 N abundance measurements. Nature 303, 658–687.
Mason, R.P., Laporte, J-M., Andres, S., 2000. Factors controlling the
bioaccumulation of mercury, methylmercury, arsenic, selenium and
cadmium by freshwater invertebrates and fish. Arch. Environ. Toxicol.
Chem. 38, 283–297.
May, T.W., Walther, M.J., Petty, J.D., Fairchild, J.F., Lucero, J.,
Delvaux, M., Manring, J., Armbruster, M., Hartman, D., 2001. An
evaluation of selenium concentrations in water, sediment, invertebrates
and fish from the Republican River basin: 1997–1999. Environ. Monit.
Assess. 72, 179–206.
McCutchan Jr., J.H., Lewis, W.M., Kendall, C., McGrath, C.C., 2003.
Variation in trophic shift for stable isotope ratios of carbon, nitrogen
and sulfur. Oikos 102, 378–390.
McDonald, L.E., Strosher, M.M., 1998. Selenium mobilization from
surface coal mining in the Elk River Basin, British Columbia. A survey
of water, sediment and biota. BC Ministry of Environment, Sept. 1998.
McDonald, L.E., Strosher, M.M., 2000. Selenium in the Elk River Basin,
British Columbia: a review of findings and discussion of implication
for assessment and management. In: Planning for End Land Use in
Mining Reclaimation. Proceedings of the 24th Annual British
Columbia Mine Reclaimation Symposium, Williams Lake, BC, June
19–24, 2000, pp. 160–173.
Merritt, R.W., Cummins, K.W., 1996. An Introduction to the Aquatic
Insects of North America, Third ed. Kendall/Hunt Publishing
Company, Dubuque, IA.
Minigawa, M., Wada, E., 1984. Stepwise enrichment of 15N along food
chains: further evidence and the relation between d15N and animal age.
Geochim. Cosmochim. Acta. 48, 1135–1140.
Minnow Environmental Incorporated (Minnow), 2003. Selenium Study of
Lentic Areas in the Elk Valley (Draft). Prepared for Elk Valley
Selenium Task Force.
Nassos, P.A., Coats, J.R., Metcalf, R.L., Brown, D.D., Hansen, L.G.,
1980. Model ecosystem, toxicity, and uptake evaluation of
75
Se-selenite. Bull. Environ. Contam. Toxicol. 24, 752–758.
NRC (National Research Council), 1989. Irrigation-Induced Water
Quality Problems: What can be Learned from the San Joaquin Valley
experience? National Research Council, Committee on Irrigation-
Induced Water Quality Problems. National Academy Press, Washing-
ton, DC.
Niimi, A.J., LaHam, Q.N., 1976. Relative toxicity of organic and
inorganic compounds of selenium to newly hatched zebrafish
(Brachydanio rerio). Can. J. Zool. 54, 501–509.
Odum, E.P., de al Cruz, A.A., 1967. Particulate organic detritus in a
Georgia salt marsh-estuarine ecosystem. In: Lanff, G.H. (Ed.),
Estuaries. American Association for the Advancement of Science
Publication, Washington, DC.
Ogle, R.S., Knight, A.W., 1996. Selenium bioaccumulation in aquatic
ecosystems: 1. Effects of sulphate on the uptake and toxicity of selenate
in Daphnia magna. Arch. Environ. Contam. Toxicol. 30, 274–279.
Ogle, R.S., Maier, K.J., Kiffney, P., Williams, M.J., Brasher, A., Melton,
L.A., Knight, A.W., 1988. Bioaccumulation of selenium in aquatic
ecosystems. Lake Reservoir Manage 4, 165–173.
O’Leary, M.H., 1984. Measurement of the isotope fractionation asso-
ciated with diffusion of carbon dioxide in aqueous solution. J. Phys.
Chem. 88, 823–825.
O’Leary, M.H., 1988. Carbon isotopes in photosynthesis. BioScience 38,
328–336.
Oremland, R.S., Steinberg, N.A., Maest, A.S., Miller, L.G., Hollibaugh,
J.T., 1990. Measurement of in situ rates of selenate removal by
dissimilatory bacterial reduction in sediments. Environ. Sci. Technol.
24, 1157–1164.
187
Ornes, W.H., Sajwan, K.S., Dosskey, M.G., Adriano, D.C., 1991.
Bioaccumulation of selenium by floating aquatic plants. Water Air
Soil Pollut. 57&58, 53–57.
Osmond, B., Valaane, N., Haslam, S.M., Uotila, P., Roksandic, Z., 1981.
Comparisons of d13C values in leaves of aquatic macrophytes from
different habitats in Britain and Finland; some implications for
photosynthetic processes in aquatic plants. Oecologia 50, 117–124.
Peterson, B.J., Fry, B., 1987. Stable isotopes in ecosystems studies. Ann.
Rev. Ecol. Sys. 18, 293–320.
Post, D.M., 2002. Using stable isotopes to estimate trophic position:
models, methods, and assumptions. Ecology 83, 703–718.
Power, M., Klein, G.M., Guiguer, K.R.R.A., Kwan, M.K.H., 2002.
Mercury accumulation in the fish community of a sub-Arctic lake in
relation to trophic position and carbon sources. J. Appl. Ecol. 39,
819–830.
Rau, G.H., 1980. Carbon-13/carbon-12 variation in subalpine lake
aquatic insects: food source implications. Can. J. Fish. Aquat. Sci.
37, 742–745.
Raven, J.A., Osborne, B.A., Johnston, A.M., 1985. Uptake of CO2 by
aquatic vegetation. Plant Cell Environ. 8, 417–425.
Rees, C.E., Jenkins, W.J., Monster, J., 1978. The sulphur isotopic
composition of ocean water sulphate. Geochim. Cosmochim. Acta 42,
377–381.
Riedel, G.F., Ferrier, D.P., Sanders, J.G., 1991. Uptake of selenium by
freshwater phytoplankton. Water Air Soil Pollut. 57–58, 23–30.
Riedel, G.F., Sanders, J.G., Gilmour, C.C., 1996. Uptake, transformation,
and impact of selenium in freshwater phytoplankton and bacterio-
plankton communities. Aquat. Microbial Ecol. 11, 43–51.
Robberecht, H., Van Grieken, R., 1982. Selenium in environmental
waters: determination, speciation and concentration levels. Talanta 29,
823–844.
Rolff, C., Broman, D., Näf, C., Zebühr, Y., 1993. Potential biomagnifica-
tion of PCDD/Fs—new possibilities for quantitative assessment using
stable isotope trophic position. Chemosphere 27, 461–468.
Rounick, J.S., Winterbourn, M.J., 1986. Stable carbon isotopes and
carbon flow in ecosystems. BioScience 36, 171–177.
Saiki, M.K., 1986. Concentrations of selenium in aquatic food-chain
organisms and fish exposed to agricultural tile drainage water. In:
Selenium and Agricultural Drainage: Implications for San Francisco
Bay and the California Environment. University of California,
Berkeley, CA, pp. 25–33.
Saiki, M.K., Lowe, T.P., 1987. Selenium in aquatic organisms from
subsurface agricultural drainage water, San Joaquin Valley, California.
Arch. Environ. Contam. Toxicol. 16, 657–670.
Saiki, M.K., Jennings, M.R., Brumbaugh, W.G., 1993. Boron, molybde-
num and selenium in aquatic food chains from the lower San Joaquin
River and its tributaries, California. Arch. Environ. Contam. Toxicol.
24, 307–319.
Sandholm, M., Oksanen, H.E., Pesomen, L., 1973. Uptake of selenium by
aquatic organisms. Limnol. Oceanogr. 18, 496–499.
Sappington, K.G., 2002. Development of aquatic life criteria for selenium:
a regulatory perspective on critical issues and research needs. Aquat.
Toxicol. 57, 101–113.
Schlekat, C.E., Dowdle, P.R., Lee, B.-L., Luoma, S.N., Oremland, R.S.,
2000. Bioavailability of particle-associated Se to the bivalve Potamo-
corbula amurensis. Environ. Sci. Technol. 34, 4505–4510.
Stephens, D., 1996. Selenium in bottom sediment from ponds at Ouray
National Wildlife Refuge, 1991, 1994. Draft Report, US Geological
Survey, Salt Lake City, UT. As cited in Hamilton et al. (2004).
Teal, J.M., 1962. Energy flow in a salt marsh ecosystem of Georgia.
Ecology 43, 614–624.
USDOI, 1998. Guidelines for Interpretation of the Biological Effects of
Selected Constituents in Biota, Water, and Sediment: Selenium. US
Department of the Interior, National Irrigation Water Quality
Program Information Report No. 3.
USEPA, 2004. Draft Aquatic Life Water Quality Criteria for Selenium—
2004. US Environmental Protection Agency, Office of Water, Office of
Science and Technology, Washington, DC November 2004.
 ARTICLE IN PRESS
188 P.L. Orr et al. / Ecotoxicology and Environmental Safety 63 (2006) 175–188
Vander Zanden, M.J., Rasmussen, J.B., 1999. Primary consumer d13C and
d15N and the trophic position of aquatic consumers. Ecology 80,
1395–1404.
Vander Zanden, M.J., Cabana, G., Rasmussen, J.B., 1997. Comparing
trophic position of freshwater fish calculated using stable isotope ratios
(d15N) and literature dietary data. Can. J. Fish. Aquat. Sci. 54,
1142–1158.
Waters, T.F., 1995. Sediment in Streams: Sources. Biological Effects and
Control. American Fisheries Society Monograph 7, Bethesda, MD 251pp.
Weiss, K.F., Ayres, J.C., Kraft, A.A., 1965. Inhibitory action of selenite
on Escherichia coli, Proteus vulgaris and Salmonella thompson.
J. Bacteriol. 90, 857–862.
Wilber, C.G., 1980. Toxicology of selenium: a review. Clin.Toxicol. 17,
171–230.
WLU (Wilfred Laurier University), 2000. Aquatic Invertebrates Illu-
strated Field Guide. http://www.wlu.ca/wwwbiol/bio305/Database/
Categories.htm. February 2000.
Woock, S.E., 1984. Accumulation of selenium by golden shiners
Notemigonus crysoleucas Hyco Reservoir N.C. cage study 1981–1982.
Carolina Power and Light Company, New Hill, NC 19pp. As cited in
Hamilton and Buhl (2003).
Wrench, J.J., 1978. Selenium metabolism in marine phytoplankters
Tetraselmis tetrathele and Dunaliella minuta. Mar. Biol. 49,
231–236.
Zawislanski, P.T., Chau, S., Mountford, H., Wong, H.C., Sears, T.C.,
2001. Accumulation of selenium and trace metals on plant
litter in a tidal marsh. Estuarine and Coastal Shelf Science 52,
589–603.