1329

Predator diet and prey adaptive responses: Can

tadpoles distinguish between predators feeding
on congeneric vs. conspecific prey?
Bastien Ferland-Raymond and Dennis L. Murray

Abstract: Predator diet can play an important role in facilitating detection of predation risk among prospective prey, and
such detection should have adaptive significance in reducing mortality in environments where not all predators confer sim-
ilar risk. In the laboratory, we tested behavioural and morphological responses of tadpoles from two congeneric frog spe-
cies (bullfrog (Rana catesbeiana Shaw, 1802) and mink frog (Rana septentrionalis Baird, 1854)) to cues from an odonate
predator (genus Aeshna Fabricius, 1775). In a separate experiment we found that both frog species had similar baseline
vulnerability to Aeshna predation, implying that species’ responses to predators feeding on conspecific vs. congeneric prey
also would be similar. Both species reduced their activity in the presence of predators feeding on tadpoles of either species
vs. those fed invertebrates (Libellulidae) or not subjected to predators (controls). Bullfrog tadpoles grew bigger than con-
trols when exposed to predators fed mink frog tadpoles only, whereas mink frogs failed to show a comparable response.
Neither species exhibited changes in shape that were attributable to predator diet. Our results suggest that closely related
frog species do not distinguish between predators feeding on conspecific vs. congeneric prey, implying that selection fa-
vours generalized antipredator responses when prey species are subject to similar predation risk.
Résumé : Le régime alimentaire des prédateurs peut jouer un rôle important en facilitant la détection du risque de préda-
tion chez les proies éventuelles; une telle détection devrait avoir une valeur adaptative en réduisant la mortalité dans les
milieux où tous les prédateurs ne représentent pas les mêmes risques. Nous avons testé, en laboratoire, les réactions com-
portementales et morphologiques des têtards de deux espèces de grenouilles appartenant au même genre (le ouaouaron
(Rana catesbeiana Shaw, 1802) et la grenouille de nord (Rana septentrionalis Baird, 1854)) à des signaux provenant d’un
odonate prédateur (le genre Aeshna Fabricius, 1775). Dans une autre expérience, nous avons trouvé que les deux espèces
de grenouilles ont des vulnérabilités de base semblables à la prédation par les Aeshna, ce qui fait croire que les réactions
de chacune des deux espèces devraient être semblables lorsque le prédateur utilise l’une ou l’autre des espèces comme
proies. Les deux espèces réduisent leur activité en présence de prédateurs en train de se nourrir de têtards de l’une ou
l’autre espèce, ce qui n’est pas le cas lorsque les prédateurs se nourrissent d’invertébrés (Libellulidae) ou lorsque les pré-
dateurs sont absents (témoins). Les têtards de ouaouarons atteignent une plus grande taille que les témoins lorsqu’ils sont
en présence de prédateurs nourris seulement de têtards de grenouilles du nord, ce qui n’est pas le cas dans la situation in-
verse chez les têtards de grenouilles du nord. Ni l’une, ni l’autre espèce ne subit de changements de forme qui pourraient
être attribuables au régime alimentaire des prédateurs. Nos résultats laissent croire que des grenouilles fortement apparent-
ées sont incapables de distinguer entre les prédateurs qui se nourrissent de proies de leur propre espèce et ceux qui con-
somment des proies appartenant au même genre qu’elles, ce qui implique que la sélection favorise des réactions
généralisées à la prédation lorsque les espèces de proies font face à des risques similaires de prédation.
[Traduit par la Rédaction]

Introduction rectly by causing changes in behaviour, morphology, or

Predator–prey relationships play an important role in de-
fining the structure and function of ecological communities.
Predation is known to directly limit, and sometimes regulate,
animal populations (Taylor 1984; Messier 1994; Oksanen et
al. 2001); however, predators also may influence prey indi-
Received 14 February 2008. Accepted 22 September 2008.
Published on the NRC Research Press Web site at cjz.nrc.ca on
7 November 2008.
B. Ferland-Raymond1 and D.L. Murray.2 Department of
Biology, Trent University, 1600 West Bank Drive,
Peterborough, ON K9J 7B8, Canada.
1Corresponding author (e-mail:
bastien.ferland-raymond@usherbrooke.ca).
2Corresponding author (e-mail: dennismurray@trentu.ca).
body condition (Harvell 1990; Lima and Dill 1990; Kats
and Dill 1998; Benard 2004). In many cases, adaptive
changes can be elicited by mere exposure to predators or
their chemical cues (Harvell 1990; Chivers and Smith 1998;
Kats and Dill 1998), and a wide variety of prey species are
known to recognize and respond to such cues (Tollrian and
Harvell 1999; Van Buskirk 2002). Predator chemical cues
are believed primarily to constitute kairomones released by
the predator and used by prospective prey to evaluate risk
(Petranka and Hayes 1998; Dicke and Grostal 2001; Blaus-
tein et al. 2004; Kusch et al. 2004). Although recent studies
reveal that predator diet affects the composition of the kair-
omone and thereby influences the magnitude and intensity
of prey responses to perceived predation risk (PPR; Belden
et al. 2000; Kiesecker et al. 2002; Li and Jackson 2005),
currently little is known about the mechanisms by which

Can. J. Zool. 86: 1329–1336 (2008) doi:10.1139/Z08-117 # 2008 NRC Canada

1330
such cues operate or the range in cues that can be detected
by a given prey species.
Predator diet should be an important determinant of prey
responses to PPR. Dietary cues from conspecific prey should
serve as a reliable index of high prey vulnerability to a
given predator (Persons et al. 2001; Wirsing et al. 2005),
and it follows that predators consuming heterospecifics
should pose a lesser threat to prospective prey than those
feeding on conspecifics (Persons and Rypstra 2000; Lind
and Cresswell 2005). Because antipredator responses of
prey can be costly (Van Buskirk 2000), selection should fa-
vour prey possessing finely tuned abilities to assess the risk
imposed by individual predators (Kusch et al. 2004). How-
ever, although comparisons showing prey discrimination of
predators feeding on conspecifics vs. heterospecifics have
been published (Chivers and Mirza 2001a), such studies
tend to compare predator diets that are vastly divergent
from a taxonomic (and presumably a chemical) perspective
(vertebrate or invertebrate: Murray and Jenkins 1999; Chiv-
ers and Mirza 2001b; Brodin et al. 2006; fruit or meat:
Nolte et al. 1994). However, prey discrimination between
predator diets is not universal (e.g., Wirsing et al. 2005;
Smee and Weissburg 2006), probably implying that not all
predators feeding on heterospecifics confer lesser mortality
risk. Indeed, if predators are opportunistic and have general-
ized feeding patterns, limited benefits should be derived
from predator diet differentiation, which should lead to non-
specific predator diet assessment (Smee and Weissburg
2006).
The level at which prey sensitivity to predator chemical
cues is sufficiently finely tuned to allow distinction between
closely related prey diets remains unclear. Studies involving
closely related predator food types have provided mixed re-
sults, with some showing the capacity for prey to distinguish
diets sharing taxonomic order (e.g., fathead minnows (Pime-
phales promelas Rafinesque, 1820) vs. green swordtails (Xi-
phophorus hellerii Heckel, 1848); Brown et al. 1995) and
others failing to show comparable effects (e.g., European
toads (Bufo bufo (L., 1758)) vs. European frogs (Rana tem-
poraria L., 1758); Laurila et al. 1997, 1998). To date the
single study evaluating predator diets among congeneric
prey (Relyea and Werner 2000) revealed that tadpoles
(wood frogs (Rana sylvatica LeConte, 1825) vs. northern
leopard frogs (Rana pipiens Schreber, 1782)) failed to dis-
criminate between prey species. However, this study only
examined prey discrimination of predator diet in one of the
two prey species, and species-specific asymmetry may exist
depending on the respective life-history pattern and natural
association with predation risk of each species. Additional
efforts clearly are needed to further elucidate mechanisms
of chemical signalling between predators and prey, and the
extent to which prey responses are finely tuned to reflect
specific predator diets.
The goal of the present study is to compare responses of
two closely related (congeneric) prey species (bullfrogs
(Rana catesbeiana Shaw, 1802) and mink frogs (Rana sep-
tentrionalis Baird, 1854)) sharing similar life-history strat-
egies and habitats to PPR elicited by predator dietary cues.
We predict that if both prey species are comparably vulner-
able to predation, then each should respond similarly to con-
specific vs. congeneric predator dietary cues; such responses
Can. J. Zool. Vol. 86, 2008
should be greater than those elicited by a heterogeneric
(family Libellulidae) predator diet. However, if prey species
differ in their baseline vulnerability to predation, then the
species suffering higher risk should exhibit less discrimina-
tion and therefore respond more strongly to all predator di-
ets.
Materials and methods
Experimental design
During summer 2006, mink frog and bullfrog egg masses
were collected in permanent wetlands near Peterborough,
Ontario (44817’N, 78819’W). Dragonfly (genus Aeshna Fab-
ricius, 1775) larvae were collected from nearby ponds.
Dragonfly larvae were present in the ponds where frog eggs
were collected, but for convenience they were collected in
ponds where they were easier to catch. Egg masses were
stored separately in 140 L bins in the laboratory until they
reached the appropriate size or stage (Gosner stage 25) for
recruitment into experiments. All the animals were cared
for in accordance with the Guide to the Care and Use of Ex-
perimental Animals by the Canadian Council on Animal
Care and the protocol was approved by the Trent University
Animal Care Committee (#06011).
First, we conducted a simple assessment of risk of tad-
poles from either frog species to predation by dragonfly
larva (predation experiment). For each species, tadpoles
from three broods were raised separately until they reached
a mean (SE) mass of 0.019 ± 0.001 g; groups of 20 tadpoles
were placed in 11 L plastic bins filled with river water and
including 50 cm of hemp rope to provide refuge cover. Tad-
poles were allowed to acclimatize for 3 h before a dragonfly
larva was introduced as a potential predator and allowed to
move freely in the tanks. Each dragonfly larva was new to
our study and used in a single experiment, and predators
were captured (and not fed) 4 days prior to the onset of the
experiment. After 12 h (from 2000 to 0800) the experiment
was terminated and remaining tadpoles were tallied to obtain
a mean survival rate per container. We conducted 24 and 29
trials for mink frog and bullfrog, respectively; differences in
levels of replication were due to seasonal disparity in the
availability of dragonfly larvae.
In a second experiment, we exposed groups of 25 tadpoles
to dragonfly-induced PPR to evaluate responses to predator
diets (diet experiment). We tested the two tadpole species
separately and used mink frog tadpoles (Gosner stage 25,
0.018 ± 0.001 g) first followed by bullfrog tadpoles (Gosner
stage 25, 0.014 ± 0.001 g). Tadpoles from three broods were
placed in 15 L experimental containers interspersed into four
treatments. Broods were kept separate to control for varia-
tion owing to parental lineage. Experimental containers
were kept in an inside laboratory at constant temperature
(20 8C) and 12 h light : 12 h dark photoperiod. Tanks were
cleaned twice a week and tadpoles were fed half a teaspoon
of boiled spinach after each cleaning.
Tadpoles were exposed to four predator diet treatments —
(1) dragonfly larva (genus Aeshna) fed mink frog tadpoles,
(2) dragonfly larva fed bullfrog tadpoles, (3) dragonfly larva
fed invertebrates (Libellulidae larvae, mostly from genus Li-
bellula L., 1758), and (4) control (no predator). Each treat-
ment was replicated 15 times, with 5 replicates per brood.
# 2008 NRC Canada
Ferland-Raymond and Murray
Predators were kept in transparent MarinaTM breeding float-
ing traps (0.6 L), allowing tadpoles to be exposed to visual
and chemical cues but preventing direct contact between
predators and prey. A wooden perching stick was added to
each predator cage, and an empty cage with a stick was
placed in the control treatment. Dragonflies were fed
~0.12 g (1 Libellulidae or 3–4 tadpoles) twice per week
after each cleaning. Feeding occurred outside the main ex-
perimental containers to ensure that no alarm cues poten-
tially released by dying prey were present in the
experimental containers; this methodology allowed only po-
tential cues associated either with predators, their diet, or a
combination of the two, to be released. The experiment was
terminated after 3 weeks and then repeated after 5 days us-
ing bullfrog tadpoles. All experimental methods and tad-
poles used were similar between species’ trials.
Behaviour
Tadpole behaviour was evaluated twice per day (morning
and afternoon), 5 days a week, for the entire duration of the
experiment. The same observer performed all behavioral
tests by checking tadpoles while standing still and moving
carefully between aquaria. All individuals that displayed
any movement within a 30 s period were counted (any tail
movement; Laurila et al. 1997) and divided by the total
number of tadpoles present in the container. This proportion
represents a snapshot measure of activity.
Morphology
At the end of the experiment, each tadpole was weighed
and photographed to evaluate the role of treatment on mor-
phology. Tadpoles were kept alive for photography, put in a
small transparent display with a ruler, and photographed at a
fixed distance from the camera lens using a Nikon D70 dig-
ital camera equipped with a Tamron 90 mm macro 1:1 lens.
Pictures with bent, blurry, or deformed tadpoles were cen-
sored from the analysis (bullfrog: 6.3%, n = 1379; mink
frog: 14.1%, n = 1241).
Statistical analysis
Survival rates from the predation experiment were ob-
tained by counting the number of tadpoles that survived at
the end of the 12 h predation trial. Species were compared
with a distribution-free t test based on bootstrapping with
1000 replicates to compensate for the lack of normality.
Data were also tested using nonparametric statistics, which
provided qualitatively similar results to the bootstrapping
exercise and thus are not presented formally herein.
In the behavioural experiment, the number of tadpoles
that were active was converted to a proportion, but propor-
tions were left untransformed because of closer adherence
to basic statistical assumptions. Difference in activity across
treatments was analysed using a repeated-measures ANOVA
with date as the repeated measure. Tukey’s HSD test was
used for further post hoc analysis. We blocked for brood
and time of day (morning or afternoon). Tests were con-
ducted using STATISTICA version 7.0 (Statsoft Inc. 2004).
Shape and size of tadpoles were analyzed from digital im-
ages (1292 for bullfrog, 1066 for mink frog). We acquired
14 landmarks (Fig. 1) using tpsDig version 2 (Rohlf 2006)
and used relative-warp analysis (Zelditch et al. 2004) to ex-
1331
amine the composite role of landmarks in determining shape
of tadpoles. Tadpole pictures from every individual con-
tainer were then isolated and superimposed using general-
ized least square Procrustes superimposition to obtain the
mean tadpole shape (consensus) for each container. Centroid
size, which consists of the square root of the sum of the
squared distance of all landmarks to the centroid, was also
calculated as a size variable. Mean size and shape per con-
tainer were used to calculate shape variables (partial warps
(PW) and uniform components) using CVAGen version 6j
(Sheets 2003). We selected relative-warp analysis over prin-
cipal components analysis (PCA) because of the former’s
superiority at discriminating size from shape (Zelditch et al.
2004) and increased power in discriminating between minor
shape differences (Birch 1997). Paired analyses for our data
revealed that PWA and PCA provided comparable results
(B. Ferland-Raymond, unpublished data).
The 22 PW and two uniform components were used as
dependent variables in a multivariate analysis of covariance
(MANCOVA) with centroid size as covariate and treatment
and brood as independent variables. Analysis was conducted
in SAS version 9.0 (SAS Institute Inc. 2002). Difference in
tadpole size between treatments was evaluated using AN-
OVA on log of centroid size, with treatment and brood serv-
ing as factors. A similar ANOVA was performed on body
mass data and produced similar results due to the close cor-
relation between body mass and size (r > 0.94, p < 0.001);
accordingly, we present only size data in the present paper.
All tests were considered significant with a < 0.05.
Results
Predation experiment
We failed to detect a difference in survival rate of bull-
frog and mink frog tadpoles (bootstrap statistic: P = 0.70).
Mean (SE) mortality rates were 43.3% ± 5.6% (n = 29) and
39.8% ± 6.7% (n = 24) for bullfrog and mink frog tadpoles,
respectively. This implies that both frog species are subject
to comparable baseline predation risk from dragonfly larvae,
and thus are expected to have comparable responses to PPR
when predators are feeding on either frog species.
Diet experiment
Behaviour
Activity of bullfrogs varied among broods (F[2,113] = 52.8,
P < 0.001); activity levels of broods ranged from 20.5% ±
0.7% to 28.8% ± 0.7%. However, we detected no relation
between time of day and bullfrog tadpole behaviour
(F[1,113] = 0.61, P = 0.44). Diet treatment elicited differential
bullfrog tadpole behaviour (F[3,113] = 21.5, P < 0.001;
Fig. 2A), with post hoc tests revealing that activity was
lower (all P < 0.001) among bullfrog tadpoles exposed to
cues from dragonfly larvae eating tadpoles of either species
compared with those feeding on invertebrates or controls
(Fig. 2A). Bullfrog tadpoles had comparable activity pat-
terns both between bullfrog and mink frog diets (P = 0.99),
as well as between control and invertebrate diets (P = 0.96).
Mink frog tadpoles responded similarly to bullfrog tad-
poles to our treatments. Both brood (F[2,113] = 6.84, P =
0.002) and time of day (F[1,113] = 49.9, P < 0.001) influ-
# 2008 NRC Canada
1332
Fig. 1. Position of tadpole landmarks (LM) used to assess morphological response of bullfrog (Rana catesbeiana) and mink frog (Rana
septentrionalis) tadpoles to predator diet. LM1: tip of the tail; LM2–LM11: perpendicular to the tail muscle at 20% distance of the tip of the
tail; LM3–LM10: perpendicular to the tail muscle at maximum tail height; LM4–LM8: perpendicular from imaginary line between LM6 and
LM13 with LM4 at the limit junction of the fin and body; LM5: eye; LM6: tip of the nose; LM7: mouth; LM9: anus; LM12–LM13–LM14:
junction between body and tail muscle at the lower end (12), muscle line (13), and upper end (14). A mink frog tadpole is displayed.
Fig. 2. Proportion of activity level of (A) bullfrog (Rana catesbei-
ana) and (B) mink frog (Rana septentrionalis) tadpoles relative to
dragonfly predator diet (C, control; L, Libellulidae; M, mink frog
tadpoles; B, bullfrog tadpoles). Values are means ± SE.
enced activity patterns. Activity ranged from 13.2% ± 0.4%
to 15.7% ± 0.5% among broods and, on average, tadpoles
were more active in the morning (16.3% ± 0.4%) than after-
noon (12.2% ± 0.3%). Diet treatment also influenced mink
frog tadpole behaviour (F[3,113] = 13.6, P < 0.001; Fig. 2B).
Post hoc tests revealed that mink frog tadpoles reduced their
activity when exposed to cues from dragonfly larvae feeding
on either tadpole diets relative to control (P < 0.001) or in-
vertebrate (P < 0.05) diets. However, tadpoles failed to dis-
Can. J. Zool. Vol. 86, 2008
criminate either between tadpole diets (P = 0.58) or between
control vs. invertebrate diets (P = 0.22).
Morphology
Bullfrog tadpole size varied among broods (F[2,54] = 64.6,
P < 0.001), with size ranging from 15.9 ± 0.3 to 17.4 ±
0.3 mm. A significant effect of treatment on size also was
observed (F[3,54] = 2.91, P = 0.043). Post hoc analysis re-
vealed that bullfrog tadpoles exposed to predators feeding
on mink frogs grew bigger than those in control treatments
(mink frog: 17.0 ± 0.3 mm; control: 16.6 ± 0.3 mm; P =
0.024; Fig. 3), but that bullfrog tadpole sizes were compara-
ble among bullfrog, mink frog, and Libellulidae treatments
(Libellulidae: 16.8 ± 0.3 mm; bullfrog: 16.8 ± 0.3 mm; P >
0.58). Bullfrog and Libellulidae diet treatments did not dif-
fer from control (P > 0.35).
Mink frog tadpoles differed in size accordingto brood
(F[2,54] = 12.0, P < 0.001), with size ranging from 16.3 ±
0.5 to 17.3 ± 0.4 mm. However, no size-related responses
to predator diet treatment were observed for mink frogs
(control: 16.5 ± 0.4 mm; Libellulidae: 16.6 ± 0.4 mm; mink
frog: 16.8 ± 0.5 mm; bullfrog: 16.7 ± 0.4 mm; F[3,54] = 1.0,
P = 0.50).
After removing the effect of size, bullfrog tadpoles were
similarly shaped in all predator diet treatments (Wilks’
l[72] = 0.123, P = 0.13). However, tadpole shape differed
among broods (Wilks’ l[48] = 0.00535, P < 0.001) and var-
ied according to tadpole size (Wilks’ l[24] = 0.292, P =
0.002). Large bullfrog tadpoles had larger tails in relation to
body size, compared with smaller tadpoles (Fig. 4A). The
shape response of mink frog tadpoles was similar to that of
bullfrogs, in that shape was comparable among predator
treatments (Wilks’ l[72] = 0.114, P = 0.12) but variable
among broods (Wilks’ l[48] = 0.0176, P < 0.001) and tad-
pole size (Wilks’ l[24] = 0.254, P < 0.001). However, larger
mink frog tadpoles did not exhibit increased tail size as did
bullfrog tadpoles. Rather, size affected mink frog tadpole
shape mostly through larger tadpoles having an upright posi-
tioning of the nose and tail tip (Fig. 4B).
Discussion
Bullfrog and mink frog tadpoles did not differ in vulner-
ability to dragonfly predation, leading us to predict that both
species would respond similarly to cues associated with
predator diet. Both species reduced their activity when ex-
# 2008 NRC Canada
Ferland-Raymond and Murray
Fig. 3. Centroid size (mm) of bullfrog (Rana catesbeiana) tadpoles
relative to dragonfly predator diet (C, control; L, Libellulidae; M,
mink frog tadpoles; B, bullfrog tadpoles). Values are means ± SE.
posed to dragonfly larvae feeding on tadpoles, but only bull-
frogs grew larger when dragonfly larvae were present. Bull-
frog size responses were limited to the mink frog treatment;
however, the absence of size difference between bullfrogs
exposed to mink vs. bullfrog tadpole diet emphasized the
lack of a discernible diet effect. These results reveal that
both tadpole species responded to predators feeding on con-
specific and congeneric prey but not on more distantly re-
lated prey, and that such responses were primarily
behavioural rather than morphological. Furthermore, the ab-
sence of response to heterogeneric prey diet compared with
the control treatment emphasizes the general importance of
predator diet in influencing adaptive responses of prey in
this system. Our results were also marked by a significant
effect of brood, which is explained by the fact that baseline
characteristics differed between parental lineages. Side anal-
ysis showed no interactions between brood and response to
PPR, thus making the response to PPR similar between
broods and the brood effect of little consequence. Collec-
tively, our findings support the hypothesis that under compa-
rable predation risk, prey should not differentiate between
predator dietary cues from closely related prey species.
These results contribute importantly to our understanding
of factors influencing perceived predation risk and prey sen-
sitivity to predator diet. They are also supported by other
studies showing prey discrimination of vastly different pred-
ator diets (Murray and Jenkins 1999; Chivers and Mirza
2001b; Brodin et al. 2006), or prey failure to exhibit differ-
entiation among congeneric diets (Rana spp.; Relyea and
Werner 2000). However, our study is the first to (i) relate
the responses of closely related predator dietary cues relative
to vulnerability to predation and (ii) document the spectrum
of symmetrical responses across a range of predator dietary
cues ranging from congeneric to vastly different taxonomic
phyla. These findings add context to our understanding of
the importance of predator dietary cues in eliciting antipre-
dator responses by prey, and are consistent with the hypoth-
esis that phylogenic proximity promotes overlapping
responses to predator dietary cues (Schoeppner and Relyea
2005). However, the above arguments are not above chal-
lenge in light of other studies revealing that highly dissimi-
lar predator diets can elicit similar antipredator responses
(Smee and Weissburg 2006) or that closely related diets
1333
Fig. 4. Shape progression of (A) bullfrog (Rana catesbeiana) and
(B) mink frog (Rana septentrionalis) tadpoles with increase in size.
Arrow origins represent the shape of small tadpoles, while arrow
heads represent the shape of large tadpoles. Vectors are magnified
10 times to emphasise differences.
may fail to cause comparable patterns (Laurila et al. 1997).
This lack of consistency among studies hints at the sheer
complexity of mechanisms eliciting prey responses to preda-
tor dietary cues, and the need to consider more broadly ex-
perimental protocols when examining such issues. Indeed,
prey discrimination of predator diets may be importantly in-
fluenced by factors such as absolute and relative mortality
risk incurred by the predator in question and structure and
function of chemical cues specific to the predator. These
need to be considered carefully in future comparisons of
prey responses to predator dietary cues.
Theory suggests that prey antipredator responses should
be especially sensitive to predators incurring high mortality
risk, and predators feeding mainly on the conspecifics
should be potentially more harmful than those feeding on
heterospecifics. Because antipredator responses to predators
with low risk may be costly and thereby maladaptive to
prey (Lima and Dill 1990; Lima 1998; Van Buskirk 2000),
the ability to recognize predator diet should be adaptive
where predators have diverse diets and pose different risks
to prey. It follows that if prey species share similar preda-
tion risk to a given predator, then prey may benefit by ‘‘pig-
gybacking’’ antipredator responses to include multiple prey
species. On the other hand, when risk consistently differs
between prey species, antipredator responses that are spe-
cific to individual prey species should be advantageous.
In our case, both tadpole species suffered similar mortal-
ity risk from dragonfly larvae and therefore the similarity in
their antipredator responses to predator diet should not be
surprising. Along the same lines, Persons et al. (2001)
showed that the wolf spider Pardosa milvina (Hentz, 1844)
avoided substrate conditioned by predators (the wolf spider
Hogna helluo (Walckenaer, 1837)) fed conspecifics; because
Hogna is known to exhibit consistent dietary preference,
Pardosa dietary sensitivity to predator dietary cues naturally
should be adaptive. On the other hand, northern quahogs
(Mercenaria mercenaria (L., 1758)) respond to blue crab
(Callinectes sapidus M.J. Rathbun, 1896) predator cues that
are independent of diet (fish vs. clam diet; Smee and Weiss-
burg 2006), likely because of the predator’s variable and op-
portunistic feeding patterns (Smee and Weissburg 2006).
Thus, we infer that mortality risk disparity among prey spe-
cies consumed by a given predator should play a role in
# 2008 NRC Canada
1334
driving evolution of predator diet sensitivity. We note that
similar mortality risk between frog species is unlikely to be
an artefact of low statistical power in our predation experi-
ment. Our a priori power analysis revealed that a reasonable
level of replication (>24 replicates per species) should pro-
vide a robust test given expected variability between species
and detectable effect sizes. However, we conducted a post
hoc equivalence test to confirm the absence of mortality
rate differences (Dixon and Pechmann 2005). Equivalence
tests require the selection of thresholds where a result would
be considered to have no biological significance. In our
case, the threshold used is the level of differences in mortal-
ity rates of bullfrogs and mink frogs that have a limited im-
pact on species evolution. By setting this threshold of
mortality rate >15%, our results are robust given our sample
sizes, whereas if we set the threshold <10%, our results are
equivocal. In light of the 3.5% difference in observed mor-
tality rates between species, and considering that a differ-
ence of 15% in mortality rate is not likely to have major
biological impact on prey survival and evolution, we con-
firm that our test had enough power to show the absence of
a trend.
It is notable that the chemical structure of predator dietary
cues themselves may elucidate the mechanism by which tad-
poles failed to differentiate between genus-related predator
diets. Indeed, the only means by which tadpole prey would
have distinguished between predator diets is if chemicals re-
leased by the predator (i.e., kairomones) were diet-specific.
To date, conflicting accounts exist on whether the source of
predator chemical cues is alarm pheromones released by
prey (Bryer et al. 2001), kairomones produced by the preda-
tor (Blaustein et al. 2004), and (or) prey leftovers from di-
gestion (Brown et al. 1995; Lefcort 1996; Stabell et al.
2003; Jacobsen and Stabell 2004). However, a mechanistic
understanding of the source of predator chemical cues may
inform our understanding of the evolution of prey sensitivity
to predator diet. Our experiment allows us to reject the pos-
sibility that tadpole responses were elicited by direct alarm
pheromones, given that predators were fed outside the ex-
perimental container yet antipredator behaviours still were
manifest. Secondly, it seems unlikely that any kairomone in
the tadpole–dragonfly system is a basic metabolic waste
emitted by the predator, because under such circumstances
tadpoles should have revealed similar responses to all preda-
tors irrespective of diet. Therefore, we suspect that in our
case the dietary cue likely was a by-product of prey diges-
tion (see also Lefcort 1996; Murray and Jenkins 1999; Dicke
and Grostal 2001; Stabell et al. 2003). This would imply
that from the evolutionary perspective, prey that share meta-
bolic similarities owing to their genetic proximity (genus-
related species) should invoke similar predator kairomones.
Therefore, if the chemical signature of the predator is the
same irrespective of diet, prey should not distinguish be-
tween predator diets. Our preliminary attempt to character-
ize chemical cues in the dragonfly–Rana system suggests
that compounds emitted by predators have similar spectral
characteristics whether predators are feeding on bullfrogs
or mink frogs, but that these differ considerably from
Libellulidae-fed predators (B. Ferland-Raymond, unpub-
lished data). This tantalizing finding highlights the need
for additional work of this nature to reveal the intricacies
Can. J. Zool. Vol. 86, 2008
of the transmission of information via chemical cues in
predator–prey systems.
Although our results indicate that prey species failed to
distinguish between predator cues from two closely related
diets, such low sensitivity may not be ubiquitous across
taxonomic groups. We found bullfrogs and mink frogs to
live sympatrically in the wild and they generally share habi-
tat requirements and have comparable life history strategies
(Stewart and Sandison 1972). Presumably, this implies that
both species are subject to comparable risk incurred by drag-
onfly and other predators. In contrast, congeneric prey that
either occupy different habitats or have disparate life-history
strategies may not necessarily piggyback their responses to
predator dietary cues even if they are taxonomically related.
However, to date apparently no study has shown that prey
can distinguish predator diets between congeneric prey, but
at the level of taxonomic order, prey differentiation of pred-
ator diet has been revealed for prey occupying nonoverlap-
ping habitats in the wild (Brown et al. 1995). This
observation implies that genus-level sensitivity to predator
diet may be especially relevant among prey species having
overlapping distributions.
Most studies evaluating the role of predator dietary cues
on antipredator responses have assessed discriminatory abil-
ity of prey without considering the broader context of preda-
tion risk differences among species under consideration
(e.g., Wilson and Lefcort 1993; Kiesecker et al. 2002; Li
and Jackson 2005). Our results suggest that selection may
have shaped antipredator responses to be fine-tuned among
related groups of prey when they share similar mortality
risk, but alternatively a general lack such fine-tuning among
closely related species also may explain our observations.
However, our findings point logically to the prediction that
congeneric prey species that are either allopatric or have dis-
similar mortality risk should have disparate responses to
predator dietary cues, with species having higher mortality
risk also exhibiting stronger responses to heterospecific diet-
ary cues. Thus, we propose that comparing PPR relative to
predator diets, across a spectrum of prey species with differ-
ences in baseline predation risk, phylogenic relatedness, and
life-history strategy, will provide a fruitful avenue for fur-
ther understanding the mechanisms underlying the ecology
and evolution of antipredator responses.
Acknowledgements
We are grateful to Tom Hossie for help in collecting ani-
mals and assisting with implementation of the experiment.
Funding for this project came from the Natural Sciences
and Engineering Research Council of Canada. This research
complies with the laws of Canada, where the research was
performed. We are grateful to anonymous reviewers for
helpful comments.
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