AQUATIC BIOLOGY
Aquat Biol
Vol. 17: 107117, 2012
doi: 10.3354/ab00465
Published online November 6
INTRODUCTION
Organic matter that supports consumers in lakes
can be produced within the ecosystem (autochtho-
nous matter) or imported into the systems from adja-
cent terrestrial habitats (allochthonous matter). In
low-productivity lakes that dominate in the boreal
ecoregion, the relative availability of these resources
is controlled by allochthonous terrestrial inputs (i.e.
leaf litter falling from riparian vegetation; Ask et al.
2009, Karlsson et al. 2009). Thus, lakes are coupled to
watershed inputs of organic matter as movements of
organic matter connect terrestrial ecosystems with
aquatic food webs (Pace et al. 2004). Moreover, be -
cause the vast majority of lakes in this region are
small and shallow and their perimeter to area ratios
are relatively high (Schindler & Scheuerell 2002),
they can be highly influenced by allochthonous
organic matter of terrestrial origin (Weidel et al.
2008, Premke et al. 2010, Solomon et al. 2011).
Littoral zones are known to receive large inputs of
allochthonous matter from riparian vegetation
(Schindler & Scheuerell 2002). Most current studies
have focused on the pelagic food webs. However, lit-
© Inter-Research 2012 · www.int-res.com*Email: patricia.glaz@uqar.ca
Determination of food sources for benthic
invertebrates and brook trout Salvelinus fontinalis
in Canadian Boreal Shield lakes using stable
isotope analysis
Patricia Glaz
1,
*
, Pascal Sirois
2
, Christian Nozais
1
1
Département de biologie et centre d’études nordiques, Université du Québec à Rimouski, Rimouski, Québec G5L 3A1, Canada
2
Laboratoire des sciences aquatiques, Département des sciences fondamentales, Université du Québec à Chicoutimi,
Chicoutimi, Québec G7H 2B1, Canada
ABSTRACT: Allochthonous inputs can be an important contribution of organic matter in lake eco-
systems. Yet, our understanding of the patterns of energy dependence of littoral invertebrates and
fish is poor. We measured carbon (δ
13
C) and nitrogen (δ
15
N) stable isotope values for primary pro-
ducers, terrestrial detritus, benthic macroinvertebrates, zooplankton and brook trout Salvelinus
fontinalis in 8 oligotrophic Canadian Boreal Shield lakes to determine food sources that support
benthic consumers and brook trout. Mixing models used to determine animal diets from stable iso-
tope analysis showed leaf litter to be the principal food source for benthic primary consumers in 6
out of 8 lakes. Brook trout derived its carbon mainly from benthic predatory macroinvertebrates in
all lakes, with a contribution ranging from 60 to 90%. Zooplankton also contributed to brook trout
diet in 3 of 8 lakes, ranging from 28 to 37%. δ
15
N increased from primary producers to consumers
at different trophic positions in all lakes. Nitrogen isotopic signatures of brook trout became more
positive with increasing fish length, indicating a change in fish foraging strategy related to size.
Our study (1) suggests that carbon from terrestrial habitat (leaf litter) may contribute significantly
to the food web of oligotrophic Canadian Boreal Shield lakes and (2) highlights the importance of
carbon originating from benthic habitats in supporting brook trout in these lakes.
KEY WORDS: Allochthonous subsidies · Benthic macroinvertebrates · Zooplankton · Brook trout ·
Diet breadth · Size-related diet shift · Stable isotopes · Boreal lakes
Resale or republication not permitted without written consent of the publisher
Aquat Biol 17: 107117, 2012
108
toral benthic pathways often play a key role in lake
production and food web dynamics (Vadeboncoeur
et al. 2002). Indeed, littoral benthic macroinverte-
brates are an important heterotrophic component of
lake ecosystems and constitute a link between basal
resources and upper trophic levels in the food web
such as fishes (Vander Zanden & Vadeboncoeur
2002, Vander Zanden et al. 2006). Knowledge of the
relative importance of allochthonous matter sources
supporting benthic macroinvertebrates and ulti-
mately fish is still poor (Carpenter et al. 2005, Solo -
mon et al. 2008). However, Cole et al. (2006) showed
that in oligotrophic lakes between 60 and 85% of
benthic macroinvertebrates used allochtonous
organic matter to meet their energy requirements.
A variety of processes at multiple trophic levels cre-
ate linkages among terrestrial, pelagic and benthic
energy pathways in lakes (Schindler & Scheue rell
2002, Vadeboncoeur et al. 2002). For instance, benthic
consumers may utilize dissolved or particulate detritus
of terrestrial origin, and fishes may consume benthic,
pelagic, or terrestrial prey (Solomon et al. 2011).
Many fishes use both benthic and pelagic habitats or
undergo ontogenic habitats shifts, integrating benthic
and pelagic food webs (Vadeboncoeur et al. 2002).
Brook trout Salvelinus fontinalis is a widely distributed
and economically important fish species of eastern
Canadian Boreal Shield lakes (Power 1980). It has
been identified as a generalist carnivore fish that
feeds mainly on benthic macroinvertebrates (Trem-
blay & Magnan 1991, Lacasse & Magnan 1992), but
also on zooplankton, terrestrial insects (Power et al.
2002, Tremblay-Rivard 2007) and fish, including
brook trout (Magnan 1988, La casse & Magnan 1992).
Recent studies have shown that stable isotope signa-
tures could be used to identify intraspecific shifts in
feeding strategies of brook trout (Power et al. 2002,
Tremblay-Rivard 2007). However, little is known
about brook trout diet and size-related diet shifts in
lakes of the Canadian Boreal Shield.
In this study, we measured naturally occurring car-
bon and nitrogen stable isotope ratios of basal re -
sources (terrestrial detritus, macrophytes, periphy-
ton, sediment organic matter), benthic organisms,
zooplankton and brook trout in 8 oligotrophic Cana-
dian Boreal Shield lakes. Using stable isotope analy-
sis we examined feeding relationships among con-
sumers. Our objectives were to: (1) determine the
ultimate sources from which benthic primary con-
sumers derive their energy in order to evaluate the
relative importance of allochthonous organic matter
for these consumers and (2) identify brook trout diet
breadth and size-related diet shifts.
MATERIALS AND METHODS
Study area
The study was conducted in the province of Que-
bec, on the Boreal Shield, in the Mistassibi River
drainage basin. The stu dy area is dominated by vir-
gin black spruce Picea mariana forests. Eight oligo-
trophic lakes with similar morphometric characteris-
tics were selected in a territory covered by pristine
boreal forest that was recently accessible by a net-
work of forestry roads (Table 1). In all lakes, the only
fish species found was brook trout, except for Lake 8,
in which brook trout coexists with white sucker
Catostomus commersoni.
Sampling
Eight lakes were sampled in July 2008. Water sam-
ples and pH measurements were taken at 5 littoral
sampling stations set uniformly around each lake. At
each sampling station, pH was measured at the sub-
surface using a YSI 556 MPS probe. Water trans-
parency was estimated with a Secchi disk at the
deepest part in each lake. Water samples (4 l) were
collected at each littoral station at 50 cm below the
surface with an Alpha bottle for the determination
of chlorophyll a (chl a), dissolved organic carbon
(DOC), total phosphorus (TP) concentrations and iso-
topic analysis of particulate organic matter (POM).
Terrestrial detritus (leaf litter, woody debris), macro-
phytes, periphyton (epilithon and epixylon), surface
sediment organic matter (SOM), benthic macro -
invertebrates, zooplankton and brook trout were
sampled for stable isotope analysis. Terrestrial detri-
tus, macro phytes and SOM were hand-collected in
the littoral zone of lakes and placed in scintillation
vials. Periphyton was collected by scraping from
rocks and woody debris in the littoral zone and
placed in scintillation vials. Benthic invertebrates
were collected in the littoral zone using a Turtox D-
net with a mesh size of 500 µm (n = 3 replicates per
station) and placed in cooler boxes. Zooplankton
samples were obtained by towing a plankton net
(mouth opening 20 cm, mesh 53 µm) mounted along-
side a boat for 5 to 10 min. Zooplankton were then
placed in Ziploc
®
bags containing water. Fish were
captured with 6 experimental gill nets, according to
the Quebec Ministry of Natural Resources and
Wildlife standard protocol (mesh sizes of 1, 1.5, 2, 2.5,
3, and 3.5 inches [25 to 90 mm]) set simultaneously
around the lake, perpendicular to the shore with the
Glaz et al.: Food web structure in Boreal Shield lakes
small mesh always set toward the shore. Gill nets
were left overnight for 12 h. All fish samples were
placed in cooler boxes and transported to the labora-
tory for processing.
Sample preparation and analyses
Water samples for the determination of chl a were
filtered (200 ml or more) onto Whatman GF/F filters.
Pigments were extracted for 24 h in 90% acetone, at
5°C in the dark without grinding. Chl a was deter-
mined using the Welschmeyer et al. (1993) method.
DOC was determined by filtering subsamples of
water through precombusted (500°C, 5 h) Whatman
GF/F filters. The filtrate was placed in glass vials with
teflon-lined caps and acidified with 25% v/v H
3
PO
4
(10 µl ml
−1
). DOC values were obtained using a TOC-
5000A analyzer (Shimadzu), following the protocol of
Whitehead et al. (2000). DOC reference standards
were produced by the Hansell’s Certified Reference
Materials (CRM) program. Total phosphorus (TP) was
measured using the molybdenum blue method (Stain-
ton et al. 1977) after autoclaving 50 ml samples with
0.5 g of potassium persulfate for 1 h at 120°C. For the
determination of isotopic signatures of POM, subsam-
ples of water were filtered through precombusted
(500°C, 5 h) Whatman GF/F filters and stored frozen
at −20°C until analysis. Terrestrial detritus, macro-
phytes, periphyton and SOM samples were cleared
and separated with the aid of a dissecting microscope.
Benthic macroinvertebrates and zooplankton samples
were identified at the order or family level using Mer-
ritt & Cummins (1996) and Edmond son (1959) respec-
tively. After identification, benthic macroinvertebrates
were frozen separately at −20°C while zooplankton
was frozen as a pool of organisms at −20°C. The total
length of each brook trout was recorded. A 2 cm
square and 1 cm deep block of dorsal white muscle
without skin sample was taken from 25 brook trout in
each lake. All muscle samples were kept frozen at
−20°C in scintillation vials until analysis.
Stable isotope analyses
POM filters, terrestrial detritus, macrophytes, peri-
phyton, SOM, whole benthic invertebrates, zoo-
plankton, and brook trout samples were dried at
60°C for 48 h. Mortar and pestle were thereafter used
to grind samples into a fine powder (except for POM
filters). Powder and filter samples were then encap-
sulated in pressed tin capsules (5 × 9 mm) and tin foil
cups (Costech Analytical Technology), respectively.
Encapsulated dry mass was approximately 1 mg for
terrestrial detritus, macrophytes, periphyton, benthic
invertebrates, zooplankton, and brook trout muscle
tissue and 3 mg for SOM. Lipid correction techniques
were not considered in our analyses because of low
carbon to nitrogen (C:N) ratio values (Vander Zan-
den & Rasmussen 1999). Previous studies showed
that lipids are lower in animals analyzed as muscle
tissue, as in our study for fish, than animals analyzed
as whole (McCutchan et al. 2003). C:N ratio values
for fish varied between 3.9 (Lake 7) and 4.7 (Lake 6)
in our study.
Analyses of stable isotopes ratios of C (δ
13
C) and N
(δ
15
N) were carried out at the Institut des sciences de
la mer (ISMER, Rimouski, Quebec, Canada) using a
COSTECH ECS 4010 Elemental Analyser coupled
with a DeltaPlus XP Isotope Ratio Mass Spectrometer
(IRMS, Thermo Electron). System control as well as
acquisition and treatment of the data were carried
out using the Isodat 2 software. Stable isotope ratios
were expressed in δ notation as parts per thousand
(‰) according to the equation:
δX = [(R
sample
R
standard
) − 1] × 1000 (1)
where X is
13
C or
15
N and R is the corresponding
13
C:
12
C or
15
N:
14
N ratios.
International standards used for the measurement
were Vienna Pee Dee Belemite (VPDB) limestone for
13
C and atmospheric nitrogen for
15
N. Regional stan-
dards for in-lab normalization regressions to deter-
mine sample δ values were anhydrous caffeine
(Sigma Chemical), Mueller Hinton Broth (Becton
Dickinson) and Nannochloropsis, respectively. These
homemade standards were calibrated once a year
using standards from the National Institute of Stan-
dards and Technology (NIST). Replicate analyses of
standards gave analytical errors (SD) of ±0.30‰ for
C and ±0.18‰ for N.
Data analyses
To estimate the relative contribution of the differ-
ent food sources to the diet of benthic invertebrates
and brook trout, we adopted a Bayesian multi-source
stable isotope mixing model, available as an open
source R package (SIAR, Parnell et al. 2010). SIAR
takes data on organism isotopes and fits a Bayesian
model to their dietary habits based on a Gaussian
likelihood with a dirichlet distribution prior mixture
on the mean. SIAR offers a number of advantages
over earlier mixing models; it can incorporate trophic
109
Aquat Biol 17: 107117, 2012
enrichment factors within the model and known C
and N concentrations dependence, assuming that for
each element, a source’s contribution is proportional
to the contributed mass times the elemental concen-
tration in that source. We assumed a trophic enrich-
ment of +1‰ for
13
C (De Niro & Epstein 1978) and
+3.4‰ for
15
N (Minagawa & Wada 1984). To assess
these trophic enrichment assumptions, we combined
organisms into main groups of consumers: zooplank-
ton, benthic primary consumers and predatory
macroinvertebrates and calculated average δ
13
C and
δ
15
N for each group.
Using Daphnia spp. as the baseline trophic level,
trophic position of consumers was calculated using
the formula given in Vander Zanden & Rasmussen
(2001):
Trophic position
consumer
=
(δ
15
N
consumer
δ
15
N
baseline
)f + 2
(2)
where δ
15
N
consumer
is the δ
15
N value of the consumer
for which the trophic position is estimated, δ
15
N
baseline
is the δ
15
N of the baseline organism, 2 is the expected
trophic position of the organism used to estimate
baseline δ
15
N, and f is the fractionation factor
between a predator and its prey which corresponds
to 1 trophic level. We used the fractionation factor of
3.4‰ as proposed by Minagawa & Wada (1984) and
generally used in studies estimating trophic position
of aquatic consumers (Vander Zanden et al. 1997).
Stable isotopes of carbon and nitrogen were also
individually plotted against fish length in each lake.
Simple linear regressions were then calculated for
each lake.
RESULTS
Physico-chemical data for the 8 lakes are outlined
in Table 1. The lakes are small (0.031 to 0.288 km
2
),
as are their catchments (0.202 to 2.895 km
2
). All the
lakes are relatively shallow (maximum depth < 9 m)
and exhibited low levels of phytoplankton biomass
(chl a) but high DOC concentrations (Table 1). This is
typical of oligotrophic Canadian Shield lakes, which
are slightly acidic, with high concentrations of humic
materials (Power et al. 2002, Glaz et al. unpubl.).
Benthic consumers and potential food sources
Mayflies (Ephemeroptera), corixids (Corixidae),
am phi pods (Amphipoda) and caddisflies (Tricho -
ptera), all of them non-predatory macroinverte-
110
Lake 1 Lake 2 Lake 3 Lake 4 Lake 5 Lake 6 Lake 7 Lake 8
Latitude (° N) 50° 25’ 44” 50° 29’ 22” 50° 23’ 13” 50° 28’ 34” 50° 30’ 9” 50° 31’ 25” 50° 30’ 40” 50° 28’ 11”
Longitude (° W) 71° 57’ 28” 71° 57’ 32” 72° 1’ 24” 71° 57’ 15” 71° 47’ 1” 71° 56’ 26” 71° 56’ 5” 71° 46’ 51”
Lake area (km
2
) 0.170 0.169 0.063 0.031 0.288 0.090 0.277 0.043
Catchment area (km
2
) 0.916 2.799 0.586 0.202 2.895 1.761 2.416 0.339
Drainage area (km
2
) 0.746 2.630 0.523 0.171 2.606 1.671 2.138 0.296
Maximum depth (m) 5 2 0.5 2 9 4.5 7.5 2
Secchi depth (m) 1.25 1.50 na
a
1.75 1.50 1.65 1.40 1.40
pH 5.92 (0.10) 5.75 (0.02) 5.94 (0.05) 5.87 (0.07) 5.92 (0.06) 5.02 (0.05) 5.62 (0.15) 5.38 (0.28)
Chl a (mg m
−3
) 0.427 (0.065) 0.390 (0.061) 0.617 (0.200) 0.363 (0.050) 0.982 (0.212) 0.546 (0.060) 0.681 (0.161) 0.486 (0.072)
DOC (mg l
−1
) 10.78 (0.57) 12.06 (0.58) 12.56 (1.01) 12.33 (0.48) 11.91 (0.73) 9.82 (0.40) 8.98 (0.43) 13.73 (1.04)
TP (µg l
−1
) 5.05 (0.26) 4.95 (0.52) 5.77 (0.50) 5.13 (1.01) 5.09 (0.70) 4.69 (0.47) 5.26 (0.70) 4.65 (0.55)
a
Not measured because lake was too shallow
Table 1. Lake characteristics of the 8 studied Canadian Boreal Shield lakes. pH, chl a, dissolved organic carbon (DOC) and total phosphorus (TP) are reported as
means (SD) over the 5 sampling stations in the photic zone. na: not available
Glaz et al.: Food web structure in Boreal Shield lakes
111
brates, were grouped as benthic primary consumers.
Leeches (Hirudinea), water mites (Hydracarina),
dragonflies (Anisoptera), damselflies (Zygoptera),
dysticids (Dysticidae) and alderflies (Sialidae), all of
them active predators, and Empidids (Empididae)
and Chironomids (Chironomidae), which are mainly
predators, were enriched in
15
N relative to primary
consumers in all lakes (Fig. 1). Consequently, these
organisms were pooled as predatory macroinverte-
brates. A mixing model was performed to evaluate
basal resource contribution to benthic primary con-
sumers. Benthic primary consumers derive their car-
bon mainly from leaf litter in all lakes (Table 2). How-
ever, in Lakes 1 and 2 epilithon and epixylon also
contributed to their diet, with mean contributions of
43 and 46% respectively (Table 2).
Brook trout diet breath
Brook trout mean carbon isotope ratios ranged
from −31.1 (Lake 8) to −26.2‰ (Lake 3) and mean
nitrogen isotopes ratios ranged from 5.6 (Lake 1) to
6.4‰ (Lake 4). Both δ
13
C and δ
15
N signatures for
brook trout were well separated from their potential
prey items in all lakes. Brook trout were enriched in
15
N relative to all other sources in all lakes (Fig. 1).
The mixing model confirmed that brook trout derive
their carbon biomass mainly from predatory benthic
macroinvertebrates in all lakes, with feasible mean
contributions ranging from 60 (Lake 2) to 90%
(Lake 3) (Table 3). However, in Lakes 1, 2 and 8, zoo-
plankton also appeared as a potentially important
food resource with feasible mean contributions of 28,
37 and 32% respectively. The mixing model also
indicated that brook trout do not depend on benthic
primary consumers for food (Table 3).
Trophic position
δ
15
N increased from primary producers to con-
sumers at different trophic positions in all lakes (Fig.
1), indicating that the heavy nitrogen isotope (
15
N)
was preferentially retained during nutrient assimila-
tion and incorporation into animal tissues. Brook
trout were enriched in
15
N by 3.77‰ relative to
predatory macroinvertebrates when all lakes were
pooled (Table 4). However, predatory macroinverte-
brates were only enriched by 2.60‰ in
15
N relative to
benthic primary consumers (Table 4).
Mean trophic position of consumers was signifi-
cantly different among the 3 main trophic groups
exa mined in all lakes (Table 5). The a posteriori
Tukey test confirmed the existence of significant dif-
ferences for all paired combinations in all lakes.
Predatory macroinvertebrates had a mean trophic
position that was significantly higher than that of
benthic primary consumers and fish had a signifi-
cantly higher trophic position than both benthic pri-
mary consumers and predatory macroinvertebrates
in all lakes (Table 5).
Size-related diet shift of brook trout
Brook trout mean length ranged from 192 (Lake 8)
to 245 mm (Lake 2). Brook trout showed great vari-
ability in δ
13
C values and they became significantly
more depleted in
13
C with increasing length in
Lakes 2, 4 and 8 (although in Lake 4 sampling size
was low, n = 8, in comparison to the other lakes, n =
25). Lakes 1, 3, 5, 6 and 7 did not show any particular
pattern in relation to increasing fish length (Fig. 2a,
Table 6). Brook trout became significantly more en-
riched in
15
N with increasing body length in all lakes
but Lake 4, and this relationship was also significant
when all lakes were pooled (Fig. 2b, Table 7).
DISCUSSION
Leaf litter appeared to be the principal carbon
source for benthic primary consumers in 7 out of our
8 lakes, suggesting a significant contribution of allo -
chthonous matter to the littoral food web. Brook trout
seemed to derive their food mainly from benthic
predatory macroinvertebrates in all lakes, although
zooplankton also contributed to brook trout diet in 3
out of 8 lakes. A size-related diet shift was also ob -
served for brook trout as they became
15
N-enriched
with increasing body length. However, no consistent
pattern was observed for δ
13
C values.
Carbon flow in lake food webs
In oligotrophic lakes, humic matter originating in
the catchment can be an important contribution of
allochthonous matter for aquatic organisms (France
1997, Glaz et al. unpubl.). Moreover, the loading of
allochthonous matter can greatly exceed autochtho-
nous primary production (Carpenter et al. 2005).
High DOC concentrations strongly influence light
penetration in lakes (Carignan et al. 2000, Karlsson
et al. 2009) and present an unfavorable environment
Aquat Biol 17: 107117, 2012
112
Fig. 1. Stable isotope signatures of carbon (δ
13
C) and nitrogen (δ
15
N) for potential food sources and brook trout for 8 lakes: (a)
Lake 1; (b) Lake 2; (c) Lake 3; (d) Lake 4; (e) Lake 5; (f) Lake 6; (g) Lake 7 and (h) Lake 8. Error bars represent the mean ± SE.
POM: particulate organic matter
Glaz et al.: Food web structure in Boreal Shield lakes
for photosynthetic fixation of carbon
in situ by autotrophic phytoplankton
(Jones 1992), limiting autochthonous
production. This is consistent with the
low chl a and high DOC values en -
countered in our studied lakes.
This study demonstrates that allo -
chthonous matter (leaf litter) is the
main food source contributor of ben-
thic primary consumers, although
benthic algae also contributed to their
diet in Lakes 1 and 2. Also, since pri-
mary consumers are prey for preda-
tory invertebrates (Merritt & Cum-
mins 1996) that are, in turn, principal
dietary items for fish, allochthonous
matter assimilated by primary con-
sumers is transferred up the food
web. Benthic invertebrates would be
then the primary link be tween terres-
trial carbon and fish. Our results sup-
port an emerging consensus that ter-
restrial and benthic support of lake
food webs can be substantial (Vade -
boncoeur et al. 2002, Premke et al.
2010, Solomon et al. 2011).
Trophic position
Our results enabled the description
of the vertical structure of the food
web using δ
15
N. Benthic primary con-
sumers, predatory macroinvertebrates
and fish had mean trophic positions of
2.25, 2.99 and 4.08 respectively. If we
consider primary producers as Tro -
phic level 1, then trophic levels of
these groups would be 1.25, 1.99 and
3.08 respectively. These results are
113
Lake Leaf litter Epilithon Epixylon Macrophytes POM
Mean Percentile Mean Percentile Mean Percentile Mean Percentile Mean Percentile
1 43 2659 abs 43 29−56 7 0−20 6 0−19
2 36 1058 10 0−20 abs 46 28−66 7 0−19
3 44 2563 3 0−9 abs 27 0−49 26 0−49
4 52 4066 17 0−35 10 0−26 13 0−31 7 0−20
5 95 9099 2 0−5 abs 1 0−4 1 0−3
6 78 6887 abs 4 0−11 6 0−17 8 0−17
7 47 2371 7 0−20 22 0−44 11 0−27 11 0−31
8 57 4570 7 0−20 10 0−25 12 0−28 13 0−28
Table 2. Bayesian mixing model (SIAR) results for benthic primary consumers in 8 lakes. Percentiles show the 1st to 99th range
of potential contribution. Sources with a necessary contribution (minimal potential contribution > 0%) in bold. abs: absent
Lake Benthic primary Predatory Fish F p
consumers macroinvertebrates
1 2.31 (0.34) 2.87 (0.41) 4.09 (0.13) 240.71 <0.001
2 2.21 (0.43) 2.97 (0.52) 3.97 (0.12) 168.10 <0.001
3 2.63 (0.33) 3.23 (0.38) 4.41 (0.14) 172.34 <0.001
4 2.25 (0.21) 3.18 (0.53) 4.22 (0.12) 90.04 <0.001
5 2.40 (0.24) 3.11 (0.30) 4.09 (0.18) 446.22 <0.001
6 2.23 (0.25) 2.91 (0.34) 4.03 (0.16) 269.27 <0.001
7 1.90 (0.26) 2.99 (0.44) 4.00 (0.16) 323.14 <0.001
8 2.08 (0.32) 2.73 (0.25) 3.85 (0.14) 324.13 <0.001
Pooled mean 2.25 (0.30) 2.99 (0.40) 4.08 (0.14)
Table 5. Mean (± SD) trophic-position estimates in all lakes and analysis of
variance (ANOVA) results comparing estimates between benthic primary con-
sumers, predatory macroinvertebrates and fish in 8 lakes. The a posteriori
Tukey test confirmed the existence of significant differences for all paired
combinations in all lakes. Significance level α = 0.05
Lake Zooplankton Benthic primary Predatory
consumers macroinvertebrates
Mean Percentile Mean Percentile Mean Percentile
1 28 20−36 3 0−9 68 58−79
2 37 30−45 2 0−7 60 51−68
3 2 0−5 8 0−17 90 80−99
4 13 0−24 14 0−35 73 48−93
5 5 0−13 8 0−19 86 74−97
6 6 0−14 7 0−17 86 76−97
7 13 0−23 6 0−15 80 70−89
8 32 23−42 3 0−8 64 55−73
Table 3. Salvelinus fontinalis. Bayesian mixing model (SIAR) results for brook trout
in 8 lakes. Percentiles show the 1st to 99th range of potential contribution. Sources
with a necessary contribution (minimal potential contribution > 0%) in bold
δ
15
N (‰) δ
13
C (‰)
Fish 5.93 (0.48) −28.70 (1.02)
Predatory macroinvertebrates 2.16 (1.20) −28.52 (1.40)
Benthic primary consumers −0.44 (0.93) −29.33 (1.45)
Zooplankton −0.57 (0.64) −32.97 (0.91)
Table 4. Mean (± SD) nitrogen (δ
15
N) and carbon (δ
13
C) stable isotope signatures
of principal groups of consumers. All lakes were pooled
Aquat Biol 17: 107117, 2012
very consistent with traditional food-chain models,
which attribute trophic positions of 1, 2 and 3 relative
to primary consumers, predatory macroinvertebrates
and fish (Vander Zanden & Rasmussen 1999). Our re-
sults are also consistent with previous isotopic studies
in which predatory macroinvertebrates had higher
15
N ratios than primary consumers and fish were en-
riched in δ
15
N relative to invertebrates (Vander Zan-
den & Rasmussen 1999, Herwig et al. 2004, Anderson
& Cabana 2007). Brook trout exhibited slightly higher
δ
15
N enrichment (3.77‰) than the typical assumed
shift of 3.4‰ per trophic level (DeNiro & Epstein 1978,
Minagawa & Wada 1984). However, Post (2002) calcu-
lated a standard deviation associated with this typical
value of ±1‰. For predatory macroinvertebrates, the
δ
15
N fractionation value was lower than expected
(2.60‰). Minagawa & Wada (1984) reported a wide
range in δ
15
N trophic enrichment values across taxa
(0 to 9‰). Moreover, δ
15
N trophic enrichment was
shown to differ between herbivores (mean = 2.5‰)
and carnivores (mean = 3.4‰) (Vander Zanden & Ras-
mussen 2001), suggesting that in our study omnivory
(feeding at more than one trophic level) may be im-
portant for predatory macroinvertebrates as well.
Brook trout, however, is corroborated as carnivorous.
Brook trout diet breadth
Stable isotopes analyses and mixing models
showed that brook trout fed primarily on benthic
predatory macroinvertebrates in the 8 Canadian
Boral Shield lakes, with a contribution ranging from
60 to 90% of fish diet. While zooplankton contributed
28 to 37% in 3 of 8 lakes, it does not appear to be a
main source of energy for fish in our lakes. Brook
trout, as with other salmonids, are visual predators
and are known to feed on the most abundant and vis-
ible food, selecting the largest prey items (Allan
114
–24
Lake 1
Lake 2
Lake 3
Lake 4
Lake 5
Lake 6
Lake 7
Lake 8
–26
–28
–30
–32
–34
–36
100 150
a
b
200 250
δ
13
C (‰)δ
15
N (‰)
300 350 400
4.5
5.0
5.5
6.0
6.5
7.0
7.5
100 150 200 250
Brook trout length (mm)
Brook trout length (mm)
300 350 400
Fig. 2. Salvelinus fontinalis. Changes in brook trout (a) car-
bon (δ
13
C) and (b) nitrogen (δ
15
N) stable isotope signatures
with increasing fish length in 8 lakes
Lake n Slope r
2
F p
1 25 0.0008 0.0003 0.0061 0.9383
2 24 −0.0081 0.4058 15.0264 0.0008**
3 25 0.0104 0.1508 4.0847 0.0551
4 8 −0.0173 0.5643 7.7710 0.0317*
5 25 0.0125 0.1436 3.8573 0.0617
6 24 −0.0072 0.0911 2.2056 0.1517
7 25 −0.0125 0.1176 3.0645 0.0934
8 24 −0.0185 0.1899 5.3916 0.0294*
All lakes 180 −0.0054 0.0150 2.7473 0.0992
Table 6. Salvelinus fontinalis. Simple regression results of
carbon stable isotope ratio (δ
13
C) against brook trout length
in 8 lakes. **p < 0.001, *p < 0.05
Lake n Slope r
2
F p
1 25 0.0117 0.3302 11.3376 0.0027**
2 24 0.0045 0.5807 30.4728 <0.0001**
3 25 0.0095 0.2344 7.0408 0.0142*
4 8 0.0086 0.3880 3.8047 0.0990
5 25 0.0152 0.7891 86.0649 <0.0001**
6 24 0.0054 0.3904 14.0915 0.0011*
7 25 0.0114 0.6383 40.5921 <0.0001**
8 24 0.0075 30.2704 8.1535 0.0092*
All lakes 180 0.0064 0.2490 59.3456 <0.0001**
Table 7. Salvelinus fontinalis. Simple regression results of
δ
15
N vs. brook trout length in 8 lakes. **p < 0.001, *p < 0.05
Glaz et al.: Food web structure in Boreal Shield lakes
1978). The larger body size of benthic invertebrates
relative to pelagic prey makes feeding on benthos
more energetically attractive to fish, increasing the
efficiency of this trophic link (Vander Zanden et al.
2006). This may explain the greater proportion of
benthic macroinvertebrates in fish diet compared to
zooplankton in our study, as also shown by other
studies (Lacasse & Magnan 1992, Power et al. 2002,
Herwig et al. 2004). Brook trout are known to coexist
in many lakes with other species, especially the
white sucker Catostomus commersonii, a specialist
benthivore (Scott & Crossman 1973). When living in
sympatry with white sucker, brook trout shift their
feeding niche from benthic macroinvertebrates to
zooplankton in response to interspecific competition
for resources (Magnan 1988, Lacasse & Magnan
1992). In this study, only Lake 8 contained both brook
trout and white sucker. In this lake, zooplankton rep-
resents 32% of the diet of brook trout according to
mixing models. This is consistent to a certain extent
with the brook trout niche shift when living in sym-
patry with white sucker. However, this may not be
the only explanation, as in Lakes 1 and 2 zooplankton
also contributed to brook trout diet (28 and 37%
respectively), but in these 2 lakes there was no pres-
ence of white sucker. Other lake characteristics may
influence brook trout planktonic feeding behavior.
In addition, other studies on brook trout have esti-
mated that they consume between 10 (Tremblay-
Rivard 2007) and up to 30% (Vander Zanden et al.
2006) of terrestrial prey. We have not taken this food
source into account and as a result may have under-
estimated the allochthonous matter source contribu-
tion to brook trout diet coming in the form of terres-
trial insects. Future studies on brook trout feeding
should consider sampling terrestrial prey as a poten-
tial food source. Invertebrate isotopic signatures vary
in the short term, possibly confounding diet compar-
isons with fish that have much slower tissue turn-
over rates. As our sampling was conducted in July,
stable isotope results would be limited to the under-
standing of the feeding behavior of brook trout dur-
ing spring or early summer.
Brook trout size-related diet shifts
Ontogenetic diet shifts and size-selective predation
are common among fish (Werner & Gilliam 1984). In
our study, δ
13
C signatures exhibited a complex pat-
tern. δ
13
C values became significantly more depleted
with increasing fish length in 3 lakes, likely indica-
ting a diet shift related to size. Other studies have
shown a significant
13
C-enrichment with increasing
fish length (Grey 2001, Xu et al. 2007), but they con-
sidered both alevin and juvenile stages, which was
not the case in our study. When all trout stages were
considered, Grey (2001) did not find a particular pat-
tern of
13
C values in relation to increasing fish length.
Furthermore, a change in δ
13
C values is indicative of
a shift in benthic or pelagic feeding (Vander Zanden
et al. 1998) and expecting that a change in diet (to a
different trophic level) can occur without a signifi-
cant change in δ
13
C may be reasonable. In contrast,
in our study, δ
15
N ratios of brook trout became more
positive with increasing fish length in 7 out of 8 lakes.
This was also shown in other studies on brook trout
(Power et al. 2002, Tremblay-Rivard 2007) and other
fish species (Grey 2001, Xu et al. 2007) and is typical
of an ontogenetic dietary shift (Power et al. 2002). As
a fish increases in length and age, it is capable of
handling larger prey items and ingesting later instars
of macroinvertebrates and terrestrial insects that fall
into the lake (Power et al. 2002).
In conclusion, our study suggests than carbon orig-
inating from terrestrial habitat, particularly leaf litter,
may significantly contribute to the food web in east-
ern Canadian Boreal Shield lakes. This study also
highlights the importance of carbon produced in
benthic habitats to support brook trout in small oligo-
trophic boreal lakes. Our stable isotope study indi-
cates that benthic macroinvertebrates are important
contributors to fish diet. It supports a growing body of
literature suggesting that benthic rather than pelagic
habitats are the primary energy pathway contribut-
ing to fish production (Vadeboncoeur et al. 2002,
Herwig et al. 2004, Weidel et al. 2008).
Acknowledgements. This is a contribution to the scientific
program of the Chaire de recherche sur les espèces aqua-
tiques exploitées. This research project was supported by a
grant from the Fonds de la recherche forestière du
Saguenay− Lac-Saint-Jean and Fonds Québécois de la
Recherche sur la Nature et les Technologies (FQRNT) to
C.N. and P.S. P.G. was supported by a PhD fellowship from
FQRNT. P. Bérubé from the Ministère des Ressources
naturelles et de la Faune du Québec also contributed to this
project. We are grateful to people who contributed to the
sampling in the field: M. Bergeron, Y. Bhérer, G. Diab and
D. Gauthier. We also thank AbitibiBowater Inc. for provid-
ing land use information and field facilities. Two anonymous
reviewers provided helpful comments on an earlier version
of the manuscript.
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117
Editorial responsibility: Asbjørn Vøllestad,
Oslo, Norway
Submitted: March 9, 2011; Accepted: July 30, 2012
Proofs received from author(s): October 16, 2012
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