Ethology 110, 635—647 (2004)

2004 Blackwell Verlag, Berlin

ISSN 0179–1613

Predation Risk Avoidance by Terrestrial Amphibians: The Role of

Prey Experience and Vulnerability to Native and Exotic Predators

D. L. Murray* , J. D. Rothà & A. J. Wirsing§

*Department of Fish and Wildlife Resources, University of Idaho, Moscow, ID,
USA;  Department of Biology, Trent University, Peterborough, ON, Canada;
àDepartment of Biology, University of Central Florida, Orlando, FL, USA;
§Department of Biological Sciences, Simon Fraser University, Burnaby, BC,
Canada

Abstract
We studied avoidance, by four amphibian prey species (Rana luteiventris,
Ambystoma macrodactylum, Pseudacris regilla, Tarichia granulosa), of chemical
cues associated with native garter snake (Thamnophis elegans) or exotic bullfrog
(R. catesbeiana) predators. We predicted that avoidance of native predators would
be most pronounced, and that prey species would differ in the intensity of their
avoidance based on relative levels of vulnerability to predators in the wild. Adult
R. luteiventris (presumably high vulnerability to predation) showed significant
avoidance of chemical cues from both predators, A. macrodactylum (intermediate
vulnerability to predation) avoided T. elegans only, while P. regilla (intermediate
vulnerability to predation) and T. granulosa (low vulnerability to predation)
showed no avoidance of either predator. We assessed if predator avoidance was
innate and/or learned by testing responses of prey having disparate levels of prior
exposure to predators. Wild-caught (presumably predator-exposed) post-meta-
morphic juvenile R. luteiventris and P. regilla avoided T. elegans cues, while
laboratory-reared (predator-naive) conspecifics did not; prior exposure to
R. catesbeiana was not related to behavioural avoidance among adult or post-
metamorphic juvenile wild-reared A. macrodactylum and P. regilla. These results
imply that (i) some but not all species of amphibian prey avoid perceived risk from
garter snake and bullfrog predators, (ii) the magnitude of this response probably
differs according to prey vulnerability to predation in the wild, and (iii) avoidance
tends to be largely learned rather than innate. Yet, the limited prevalence and
intensity of amphibian responses to predation risk observed herein may be
indicative of either a relatively weak predator–prey relationship and/or the limited
importance of predator chemical cues in this particular system.
Corresponding author: D. L. Murray, Department of Biology, Trent Univer-
sity, Peterborough, ON, Canada, K9J 7B8. E-mail: dennismurray@trentu.ca

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636 D. L. Murray, J. D. Roth & A. J. Wirsing

Introduction
Prey animals may exhibit a suite of behavioural responses following exposure
to predators, including direct displacement and alterations in movement or
activity patterns (e.g. Lima & Dill 1990; Sih et al. 1990). Recent research has
shown that the expression of such behaviours can be mediated by chemical cues
associated with a predator; in systems where predator chemical cues can be used
to forecast a predatory encounter, adaptive responses by prey to such cues should
be favoured from an evolutionary standpoint (Vermeij 1994; Abrams 2000).
Moreover, such responses should be related to the magnitude of predation risk
faced by the prey species, as well as the length of the prey’s exposure to predation
risk. It follows that in the case of predator–prey relationships where either
predation risk is presently high or where predator-induced selection has existed
for a long time, prey behavioural responses to perceived predation risk should be
substantive.
Prey learning also may shape prey responses to predation risk, thereby
further complicating the dynamic interaction between predator and prey. In
theory, the extent of predation upon the youngest age classes and the likelihood
of prey surviving an initial predator encounter, and responding adaptively
following subsequent encounters, should in large part determine whether a
particular anti-predator response is innate or learned. Because many predator–
prey relationships are both complex and variable, for some prey species
predator avoidance may possess both innate and learned components. It
follows that the disparity in both the ecology and evolution of predator–prey
interactions across taxonomic groups may lead to differential ontogeny and
intensity of anti-predator behaviour across predator and prey species (e.g.
Dickman 1992; Jedrzejewski et al. 1993).
Many aquatic amphibian species exhibit predator avoidance that is
mediated by chemical cues (e.g. Peterson et al. 1992; Kats & Dill 1998), likely
because predators and prey are often found in close proximity and exposure to
predator cues tends to be intense. Limited evidence exists for similar patterns of
recognition in terrestrial environments (e.g. Cupp 1994; Madison et al. 1999;
Murray & Jenkins 1999). In the northwestern US, pond amphibians are preyed
upon extensively by native garter snake predators (Thamnophis elegans,
T. sirtalis) (Nussbaum et al. 1983); garter snakes (or their chemical cues) are
known to elicit a variety of avoidance behaviours among amphibian prey
(Madison et al. 1999; Murray & Jenkins 1999; Belden et al. 2000). The bullfrog
(Rana catesbeiana), introduced recently (last 50 yr) from the eastern US to many
western ponds (Nussbaum et al. 1983), is also known to prey on juvenile and
adult forms of many species of western frogs and salamanders (Bury & Whelan
1984; Schwalbe & Rosen 1988), and also may compete with native amphibians
for space or food (Bury & Whelan 1984). (Herein we refer to bullfrogs as
amphibian ÔpredatorsÕ, recognizing that if they also serve as competitors,
amphibian avoidance behaviour should be accentuated.) Both adult and larval
bullfrogs have been shown to elicit avoidance in a variety of aquatic amphibians
 Predation Risk Avoidance by Terrestrial Amphibians 637
(Kiesecker & Blaustein 1997; Kiesecker et al. 2001), implying that their potential
impact on such species may be substantive, perhaps even contributing to
declines in some amphibian populations (e.g. Hayes & Jennings 1986; Kiesecker
& Blaustein 1997; Kupferberg 1997).
Post-metamorphic amphibians may be less susceptible to predation by garter
snakes and bullfrogs than their larval counterparts because they avoid aquatic/
riparian habitats during most of the year. Thus, while aquatic amphibian larvae
should avoid chemical cues from these two predators, expression of such
responses may be absent among some adult amphibians because they confer little
selective advantage (i.e. in cases where sympatry between adult amphibians and
their predators is limited). Similarly, predator avoidance may not be beneficial in
toxic amphibian species.
We assessed behavioural responses of spotted frogs (R. luteiventris), long-
toed salamanders (Ambystoma macrodactylum), Pacific treefrogs (Pseudacris (or
Hyla) regilla), and rough-skinned newts (Tarichia granulosa) to chemical cues
associated with native (garter snake) vs. exotic (bullfrog) predators. We
hypothesized that predator avoidance would be associated with relative vulner-
ability to predation by different prey species. We predicted that (1) adult spotted
frogs, which not only occupy riparian habitat throughout the spring–fall
(Nussbaum et al. 1983) and are therefore in close proximity to both garter
snakes and bullfrogs for much of the year, but also have been observed by us
being consumed by both predators, should avoid chemical cues from both
predator species. Adult salamanders and treefrogs should not avoid such predator
cues as strongly because they occupy subterranean (salamanders) or arboreal
(treefrogs) habitats (i.e. spatial refugia) during most of the spring–fall. Newts are
toxic to both garter snakes and bullfrogs (see Brodie 1968; Macartney & Gregory
1981), and thus should gain the least benefit from any predator avoidance. Thus,
the four amphibian species should exhibit a range in prey avoidance behaviour
that is based on their susceptibility to snake and bullfrog predation. (2) The
disparity in past exposure of western larval amphibians to the two predators
suggests that (i) avoidance should be more intensive towards snake cues, and that
(ii) predator avoidance should be strongest among post-metamorphic juvenile
stages than adults (i.e. largely innate).

Methods
Collection and Maintenance of Study Animals
All animals were handled in accordance with protocol number 9053, which
was approved by the University of Idaho Animal Care and Use Committee.
Free-ranging adult salamanders, newts, and frogs were captured by hand,
dipnet, or minnow trap from breeding ponds near Moscow, Idaho (49N,
117W). Adult salamanders and treefrogs were captured during the breeding
season (March–May), spotted frogs were captured during April–August, while
newts were captured during August–September. Post-metamorphic juvenile
638 D. L. Murray, J. D. Roth & A. J. Wirsing

frogs, salamanders, and treefrogs were obtained near pond edges a few days
before or after metamorphosis; newly transformed juvenile newts were not
collected. Overall, amphibians were obtained from 10 different ponds, only two
that are also known to be occupied by resident populations of bullfrogs
(separate groups of long-toed salamanders and treefrogs were captured from
ponds with and without bullfrogs; spotted frogs and newts were available from
ponds lacking bullfrogs only); garter snakes were observed in/near all ponds.
Because ponds with bullfrogs were separated by a minimum of 8 km from
those without bullfrogs, we considered all amphibian populations to be discrete
with respect to exposure to bullfrogs. All amphibians were housed in plastic
containers in a coldroom (5C) for 1–3 d prior to the behavioural trials;
animals were allowed to acclimate to room temperature for several hours prior
to the start of trials.
To evaluate the extent to which amphibian responses to garter snake
predation risk were innate and/or learned, we raised frog tadpoles and
salamander larvae in the laboratory and subjected them to snake chemical cues
in behavioural trials, post-metamorphosis. Gastrula-stage amphibian egg masses
were removed from ponds in spring and housed in 2-gallon plastic containers
containing pond water and receiving continuous aeration. Hatched larvae were
fed ad lib until metamorphosis (salamanders were fed brine shrimp and
oligochaete worms; frogs were fed tropical fish food). Post-metamorphic juveniles
were removed from containers after transformation and used within 3 d in
behavioural trials (July–September). To evaluate the effect of prior exposure on
amphibian avoidance of bullfrog cues, post-metamorphic juvenile and adult
salamanders and treefrogs from ponds with vs. without bullfrogs were captured
and subjected to behavioural trials.
Wandering garter snakes (n ¼ 23) were captured by hand and housed
individually in 10-gallon tanks, while bullfrogs (n ¼ 12) were either captured
from ponds or obtained commercially, and housed in 50-gallon plastic storage
bins. In addition, to test the possibility that newts avoided cues from snakes
that were toxin-resistant (see below), five common garter snakes (T. sirtalis)
were captured and housed in conditions similar to that of T. elegans. Snakes
and bullfrogs were kept in the laboratory year-round, and fed earthworms
(Lumbricus spp.) or chicken hearts, every 3–4 d. Predators were fed
earthworms rather than conspecific prey specifically to provide a neutral diet
that would not bias interspecific comparisons of prey responses; because prey
avoidance tends to be accentuated when conspecifics are fed to predators, the
intensity of prey responses reported herein may be considered as conservative.
All animals were maintained on a 12.00 : 12.00 hours (light : dark) photo-
period at approximately 20C. Prior to behavioural trials, paper towels were
left in the predator tanks/storage bins for 24–36 h, and then used as substrate
in experiments. Predators were made to fast for a minimum of 4 d prior to
being used in experiments; however, any faecal material that accumulated on
paper towels during the treatment period was removed prior to behavioural
trials.
 Predation Risk Avoidance by Terrestrial Amphibians 639
Behavioural Trials
Our methods for evaluating anti-predator behaviour were similar to those
used by Cupp (1994) and Murray & Jenkins (1999). Amphibians were placed in
24 cm · 45 cm behavioural arenas with a central line drawn on the bottom and
one paper towel (substrate) on each side of the arena. Most trials were conducted
under a fume hood to prevent cross-contamination of predator cues. Amphibian
behaviour was observed for 120 consecutive minutes in a laboratory under
fluorescent lighting (e.g. Chivers et al. 1995), and all behavioural trials were
conducted at room temperature between 10.00 and 15.00 hours. The two paper
towel substrates in each arena were separated by 2 cm to avoid diffusion of
chemical cues between them. At the start of each trial, amphibians were released
on the centre line; every 2 min their location in the experimental chamber (side A
or side B) was recorded. After 30, 60 and 90 min, we rotated the arena 180. To
minimize the potential for amphibian dehydration during trials, we misted paper
towels with dechlorinated water and covered experimental chambers with
plexiglass.
Wild-caught adult and post-metamorphic juvenile amphibians and laborat-
ory-reared juvenile amphibians were subject to predation risk treatments from
wandering garter snakes and bullfrogs vs. untreated (control) substrates. Because
we were unable to obtain newt eggs or juveniles, this species was excluded from
such trials. To evaluate the ability of newts to discern between toxin-resistant vs.
potentially non-resistant snake predators, we tested adult newts in behavioural
trials consisting of common garter snake chemical cues (common garter snakes
exhibit greater resistance to newt toxin than do wandering garter snakes, see
Brodie & Brodie 1999). Baseline information on amphibian movement patterns in
the absence of predator cues was determined in trials consisting of control
substrate on both sides of the arena.

Statistical Analysis
In sum, we conducted a total of 520 behavioural trials evaluating amphibian
anti-predator responses, with 160 trials being undertaken on salamanders, 160 on
treefrogs, 120 on spotted frogs, and 80 on newts. An individual amphibian was
considered as the experimental unit and thus was used in a single trial, except that
11 adult treefrogs and 20 adult newts were used in two trials each. Because for
each individual used in multiple trials the second trial constituted exposure to
control vs. control substrate (to determine baseline activity patterns), and given
the relatively large number of predators used to condition the substrate, we felt
that this level of pseudoreplication was unlikely to have confounded our results
(see also Ramı́rez et al. 2000). While our experimental lighting regime and
intensity of predator cues were comparable with those used in previous studies, we
determined if our results were influenced by these procedures by comparing the
behaviour of adult salamanders (subject to snake vs. control treatments) when
(i) fluorescent lights were on vs. off, and (ii) paper towels had been left in snake
640 D. L. Murray, J. D. Roth & A. J. Wirsing

tanks for 1 vs. 3 d. These tests involved conducting an additional 40 behavioural

trials (20 with lights off, 20 with 3-d-old paper towels) and comparing the results
with a set of 20 trials using standard experimental protocols (i.e. lights on and 1-d-
old paper towels) and involving snake cues. We found that neither lighting level
(F1,38 ¼ 0.985, p ¼ 0.32), nor duration of exposure to paper towels (F1,38 ¼
0.084, p ¼ 0.77) affected salamander behavioural responses to predator cues,
implying that our results are unlikely to have been confounded specifically by our
experimental protocol.
We sampled 20 individuals for each permutation of our experiment; this
standardized approach eliminated potential effects of sample size variability on
statistical power. Murray & Jenkins (1999) showed that a similar sample size was
adequate to detect avoidance of garter snake cues by red-backed salamanders
(Plethodon cinerus) under a variety of experimental conditions, including a
standard earthworm diet for snakes. Considering that a 60–65% use of the
control substrate (contrary to the null of 50%) might represent biologically
significant avoidance of predator cues (keeping in mind that the neutral predator
diet likely reduced the level of anti-predator stimulus), we conducted a power
analysis using our sample size (n ¼ 20) and the mean standard deviation observed
across our main trials (SD ¼ 0.218; see Table 1). We found that for a ¼ 0.05,
statistical power for a single-sample t-test was 0.50 for a population mean of 60%,
and 0.83 for a mean of 65%. Thus, we considered that our experimental protocol
provided reasonable statistical power for detecting predator avoidance in the
species under study.
We used overall percent time that subjects spent on the control side of the
arena to measure antipredator behaviour. Because all of our sample means for use

Table 1: Percent use of control substrates by four amphibian prey species during experi-
ments evaluating the effect of predator chemical cues on avoidance behaviour

Species Treatment Percentage SD t-value p-value

R. luteiventris Control 0.506 0.151 0.176 0.86

A. macrodactylum Control 0.482 0.187 0.481 0.63
P. regilla Control 0.506 0.071 0.347 0.73
T. granulosa Control 0.466 0.257 0.591 0.56
R. luteiventris T. elegans 0.576 0.100 3.392 0.003
A. macrodactylum T. elegans 0.608 0.173 2.797 0.012
P. regilla T. elegans 0.547 0.221 0.960 0.35
T. granulosa T. elegans 0.492 0.228 0.148 0.88
R. luteiventris R. catesbeiana 0.640 0.116 5.370 <0.001
A. macrodactylum R. catesbeiana 0.505 0.270 0.078 0.90
P. regilla R. catesebiana 0.426 0.252 1.316 0.20
T. granulosa R. catesbeiana 0.549 0.233 0.942 0.36
T. granulosa T. sirtalis 0.475 0.206 0.539 0.60

All prey animals used in trials involving bullfrog cues were captured from ponds lacking
bullfrogs. Each trial involved 20 prey animals and analyses were performed using one-
sample t-tests, which evaluated the hypothesis of random (i.e. 50 : 50) use of substrates.
 Predation Risk Avoidance by Terrestrial Amphibians 641
of control substrate ranged between 25–75% and the distribution of data from all
trials approached normality, percentage data were left untransformed and
parametric tests were used in analyses. Percent avoidance of predator chemical
cues was analysed using a one-sample t-test (Zar 1984) against the null hypothesis
of no avoidance (i.e. 50% use of each side). anova was used to compare the effect
of specific treatments on amphibian avoidance. While we recognize that multiple
tests can inflate the chances of type I statistical error, we reject the concern that
Bonferonni adjustments should be used to correct our p-values in the above tests.
Each statistical test that used previously analysed data addressed a unique
hypothesis pertaining to predator species (exotic vs. native) or response type
(innate vs. learned), while each data set used in such analyses was a composite of
those used in previous tests.

Results
Avoidance of Predator Cues by Adult Amphibians
In control vs. control trials, the four prey species showed no selection for
specific sides of the experimental chamber, indicating that there were no
directional biases inherent in our experimental procedure (Table 1). We tested
for amphibian avoidance of snake and bullfrog chemical cues, where all
individuals tested against bullfrogs were from ponds lacking these predators.
Consistent with our first prediction, spotted frogs were the only prey species
where adults avoided chemical cues from both predators, with subjects spending
on average 58% of the trial away from wandering garter snake cues, and 64%
from bullfrog cues (Table 1). The intensity of avoidance manifested by adult
spotted frogs did not differ significantly between garter snake and bullfrog
predator cues (F1,38 ¼ 3.396, p ¼ 0.073). In contrast, adult long-toed salaman-
ders were found to avoid wandering garter snake cues but not those of bullfrogs
(Table 1), but intensity of salamander avoidance did not differ relative to the two
species of predators (F1,38 ¼ 0.202, p ¼ 0.65). Neither adult treefrogs nor newts
avoided chemical cues from either of the two main predator species (Table 1).
Furthermore, newts did not avoid cues from common garter snakes (Table 1),
and the pattern of substrate usage was similar for trials with both snake species
(F1,38 ¼ 0.064, p ¼ 0.80).

Innate vs. Learned Responses

We compared the responses of laboratory-reared (predator naive) vs. wild-
caught (ostensibly predator-exposed) spotted frogs, salamanders, and treefrogs,
with wandering garter snake cues. Wild-caught juvenile spotted frogs and
treefrogs exhibited snake avoidance, while laboratory-reared conspecifics did not
(Table 2). However, we found that for both spotted frogs (F1,38 ¼ 0.002, p ¼
0.99) and treefrogs (F1,38 ¼ 0.542, p ¼ 0.47), the intensity of avoidance did not
differ between laboratory-reared vs. wild-caught recent metamorphs. Moreover,
642 D. L. Murray, J. D. Roth & A. J. Wirsing

Table 2: Percent use of control substrates by three amphibian prey species in experiments
evaluating the effect of prey experience on avoidance of predator chemical cues

Species Treatment Age Origin Percentage SD t-value p-value

R. luteiventris T. elegans J Lab 0.556 0.190 1.315 0.20

R. luteiventris T. elegans J Wild 0.556 0.112 2.238 0.032
A. macrodactylum T. elegans J Lab 0.553 0.253 0.923 0.37
A. macrodactylum T. elegans J Wild 0.522 0.277 0.358 0.72
P. regilla T. elegans J Lab 0.561 0.256 1.070 0.30
P. regilla T. elegans J Wild 0.616 0.120 2.462 0.024
R. luteiventris B. catesbeiana J Non-BF 0.535 0.168 1.304 0.20
R. luteiventris B. catesbeiana A Non-BF 0.640 0.116 5.370 0.001
A. macrodactylum B. catesbeiana J Non-BF 0.553 0.180 1.324 0.20
A. macrodactylum B. catesbeiana J BF 0.508 0.313 0.002 1.00
A. macrodactylum B. catesbeiana A Non-BF 0.505 0.270 0.078 0.94
A. macrodactylum B. catesbeiana A BF 0.605 0.319 1.476 0.16
P. regilla B. catesbeiana J Non-BF 0.576 0.337 0.882 0.39
P. regilla B. catesbeiana J BF 0.482 0.166 0.493 0.63
P. regilla B. catesbeiana A Non-BF 0.426 0.252 1.316 0.20
P. regilla B. catesbeiana A BF 0.564 0.257 1.118 0.28

Each trial involved 20 prey animals and analyses were performed using one-sample t-tests,
which tested the hypothesis of random (i.e. 50 : 50) use of substrates. Adult and newly
transformed (juvenile) animals served as test subjects. Laboratory-reared and wild-caught
juveniles were used in tests involving snake cues, whereas prey animals from bullfrog (BF)
vs. non-bullfrog (non-BF) ponds were used in bullfrog trials. Spotted frogs were una-
vailable from non-bullfrog ponds and thus not tested.

the significant predator avoidance demonstrated by juveniles did not differ from
the behavioural patterns reported for adults of either anuran species (spotted frog:
F1,38 ¼ 0.375, p ¼ 0.55; treefrog: F1,38 ¼ 1.008, p ¼ 0.32). In contrast, neither
laboratory- nor wild-reared juvenile salamanders exhibited predator avoidance
when subject to snake cues (Table 2).
Bullfrog-naive adult spotted frogs (no spotted frogs were available from
bullfrog ponds) avoided bullfrog cues, but we failed to detect a similar response
among recent metamorphs (Table 2). Next, we compared behavioural responses
of adult and juvenile salamanders and treefrogs captured from ponds with and
without bullfrogs. For adults of both species, we found that individuals failed to
avoid bullfrog cues, irrespective of whether they were from bullfrog or non-
bullfrog ponds (Table 2). Intensity of avoidance did not differ between bullfrog-
exposed and -naive adult salamanders (F1,38 ¼ 1.162, p ¼ 0.29) and treefrogs
(F1,38 ¼ 2.957, p ¼ 0.094). Similarly, we failed to detect avoidance among recent
metamorphs of either species from bullfrog or non-bullfrog ponds (Table 2), and
the intensity of any potential avoidance behaviour also was similar between
bullfrog/non-bullfrog capture sites (salamander: F1,38 ¼ 0.323, p ¼ 0.57; treefrog:
F1,38 ¼ 1.018, p ¼ 0.32).
 Predation Risk Avoidance by Terrestrial Amphibians 643
Discussion
The results of this study generally support the contention that predator
avoidance behaviour is shaped by vulnerability to predation by a given predator
species and, in at least some cases, prey experience. Adult spotted frogs (high
predation risk) avoided chemical cues associated with both garter snake and
bullfrog predators, long-toed salamanders (intermediate predation risk) avoided
snake cues only, while treefrogs (intermediate predation risk) and newts (low
predation risk) did not avoid predator cues (Table 1). For juvenile spotted frogs
and treefrogs, avoidance of snake cues appeared to be largely learned (but lost
among adult treefrogs), whereas no positive effects of prior exposure to bullfrogs
were observed for salamanders or treefrogs (Table 2).
Prey avoidance of predator chemical cues has been demonstrated across a
wide range of taxa (see Kats & Dill 1998 for review), and a variety of evolutionary
arguments have been used to explain the genesis of such behaviour. The ability to
recognize potential predators through chemical cues clearly has adaptive value
insofar as it may decrease the probability of encounter with, and subsequent
capture by, a predator. However, if such behaviour is so generalized that it is
elicited by a wide range of heterospecifics (including non-predators or
opportunistic predators), then avoidance could be less effective from the cost–
benefit standpoint (Lima & Dill 1990; Kiesecker et al. 1996). It follows that prey
behavioural responses should be largely suited to offset risk elicited from a
particular predator only where such risk is both predictable and associated with a
high probability of mortality. Accordingly, prey species should exhibit predator-
specific variability in their response to perceived predation risk. However, if the
predator–prey relationship was shaped principally by past evolutionary history,
or if adaptive response to predation risk is constrained by conflicting selective
forces opposing such a response, then the relationship between predator
avoidance and perceived predation risk should be weakened.
Spotted frogs were the only prey species where adults avoided cues from both
snakes and bullfrogs. This observation is consistent with our first prediction,
given that (1) spotted frogs and both predators have broadly overlapping habitat
use patterns in the wild (i.e. riparian areas, Nussbaum et al. 1983), and (2) this
species is known to be a palatable prey item for garter snakes and bullfrogs in our
study area. The more limited anti-predator response of long-toed salamanders
may stem from the tendency of this species to occupy subterranean habitat for
much of the year, where the risk of predation by garter snakes and bullfrogs is
likely reduced. The avoidance of snake cues only in this species could be related to
the higher importance of snakes as a salamander predator and/or a longer-term
interaction between salamanders and snakes than between salamanders and
bullfrogs. Regrettably, these latter scenarios cannot be assessed thoroughly, for
while bullfrogs are suspected predators of several western anuran species (Ehrlich
1979; Kiesecker & Blaustein 1997; Kupferberg 1997), studies comparing the
efficacy of the bullfrog as an amphibian predator relative to the garter snake in the
western United States have yet to be conducted (see Ferguson 1961; Nussbaum
644 D. L. Murray, J. D. Roth & A. J. Wirsing

et al. 1983; Beneski et al. 1986). For treefrogs, minimal habitat overlap with the
predators in question (and thus low predation risk) probably contributed to their
failure to avoid snake and bullfrog cues. However, it is notable that Chivers et al.
(2001) observed that predator-exposed treefrogs avoided cues associated with
bullfrogs, and the disparity between their research findings and ours may be
related to spatial differences in bullfrog predation risk or other local factors
affecting trade-offs in the predator–prey interaction. Finally, we would argue that
the lack of antipredatory behaviour among the newts tested suggests that the
toxicity, and consequent low palatability of this species, has removed predator-
mediated selective pressure to avoid predator chemical cues.
Predation risk avoidance has been shown to possess both innate and learned
components depending on a variety of factors related to predator and prey life
history. Studies using laboratory-reared prey residing in aquatic environments
have shown that anti-predatory behaviour has a strong genetic basis (e.g. Sih &
Kats 1994), but that aspects of the response may be learned (Semlitsch & Reyer
1992; Chivers et al. 1995; Woody & Mathis 1998; Wildy & Blaustein 2001). We
found that predator-naive juvenile amphibians failed to show significant
avoidance of snakes, while predator-exposed juveniles from two species did
exhibit such avoidance. While this finding suggests that amphibian responses to
predation risk are shaped by prior exposure and thus are learned, it is notable that
the actual intensity of snake avoidance was modest and that laboratory-reared
individuals also tended to avoid snakes, albeit only qualitatively (Table 2).
Accordingly, it remains possible that to some extent predator avoidance was of
innate origin and perhaps refined or intensified following amphibian maturation
and/or direct exposure to predation risk. This result is corroborated by the
observed avoidance of bullfrog cues by bullfrog-naive adult spotted frogs, and the
fact that a similar significant response was absent among newly transformed
juveniles. Yet, because in the present study there was no clear effect of prior
exposure to bullfrogs on amphibian responses, the short duration of bullfrog
interactions with native amphibians, and/or the modest mortality risk caused by
bullfrogs in the wild, could have precluded the significant expression of predator
avoidance. Alternatively, amphibian responses to perceived risk may simply differ
fundamentally between snake and bullfrog predators.
The idea that prey animals avoid perceived predation risk is widespread in
ecology, and the vast majority of studies addressing this topic have demonstrated
such avoidance (Lima & Dill 1990; Kats & Dill 1998). However, several published
studies have failed to demonstrate a substantive prey response (e.g. Heinen 1994;
Wolff & Davis-Born 1997; Kullberg & Lind 2002), and others have recently
shown that such responses are more variable than previously thought (e.g. Hazlett
& Schoolmaster 1998; Brown & Warburton 1999; Martı́n et al. 2003; Mathis et al.
2003). Thus, while the frequency with which predator avoidance has been
demonstrated may reflect the actual prevalence of this phenomenon across taxa,
this trend may instead be symptomatic of a publication bias favouring significant
results (see Csada et al. 1996). Here, having invoked a robust study design likely
to furnish non-significant results for species showing no response, we report a low
 Predation Risk Avoidance by Terrestrial Amphibians 645
intensity of predator avoidance, even in comparison with studies on similar
species also involving a ÔneutralÕ predator diet (e.g. Cupp 1994; Madison et al.
1999; Murray & Jenkins 1999). By implication, we suggest that prey responses to
perceived predation risk are subject to considerable taxonomic, geographic, and/
or environmental variation that may be related to habitat-specific risk levels
(Krupa & Sih 1999), and that the limited prevalence and intensity of behavioural
responses reported herein may be indicative of either relatively weak predator–
prey relationships and/or the limited importance of predator chemical cues in this
system. However, because most attempts to evaluate prey responses to predator
cues have occurred in the laboratory, where the intensity of predator stimuli may
be unnaturally high and the potential for abnormal prey behaviour is substantive,
it remains necessary to further assess how predator cues affect the fitness of free-
ranging prey. Future studies need to assess the linkage between mortality risk
incurred by a predator, how such risk affects prey behaviour, and how
behavioural changes influence prey life history, survival, and production (e.g.
Chivers et al. 1999; Maerz et al. 2001; Mirza & Chivers 2001; Gazdewich &
Chivers 2002; Kiesecker et al. 2002), and ultimately, predator–prey population
dynamics (see Vet 1999). Such studies are particularly needed in relation to
varying predator diets and instances (such as ours) where avoidance responses
appear to be biologically modest. Our results lead to the prediction that for most
species, perceived predation risk will be linked to prey avoidance behaviour, but
that such a linkage will be adjusted by other factors related to the life history of
the prey species.

Acknowledgements
We are very grateful to B. Barton, G. Bjornstrom, J. Brown, C. Holmberg, J. Nelson, D. Speten,
and C. Strobl for assistance in the lab and in the field. We thank S. Forbes, J. Oakleaf, and two
anonymous reviewers for helpful comments on a previous draft of the paper. The study was funded by
the University of Idaho Seed Grant program.

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Received: July 20, 2003

Initial acceptance: May 17, 2004

Final acceptance: May 17, 2004 (S. Forbes)