Oecologia (2006) 147: 585–595
DOI 10.1007/s00442-005-0301-3
POPULATION ECOLOGY
Anssi Laurila Æ Susanna Pakkasmaa Æ Juha Merilä
Population divergence in growth rate and antipredator defences
in Rana arvalis
Received: 6 April 2005 / Accepted: 1 November 2005 / Published online: 2 December 2005

Springer-Verlag 2005
Abstract Growth and development rates often differ
among populations of the same species, yet the factors
maintaining this differentiation are not well understood.
We investigated the antipredator defences and their
efficiency in two moor frog Rana arvalis populations
differing in growth and development rates by raising
tadpoles in outdoor containers in the nonlethal presence
and absence of three different predators (newt, fish,
dragonfly larva), and by estimating tadpole survival in
the presence of free-ranging predators in a laboratory
experiment. Young tadpoles in both populations
reduced activity in the presence of predators and
increased hiding behaviour in the presence of newt and
fish. Older tadpoles from the slow-growing Gotland
population (G) had stronger hiding behaviour and lower
activity in all treatments than tadpoles from the fast-
growing Uppland population (U). However, both pop-
ulations showed a plastic behavioural response in terms
of reduced activity. The populations differed in induced
morphological defences especially in response to fish. G
tadpoles responded with relatively long and deep body,
short tail and shallow tail muscle, whereas the responses
in U tadpoles were often the opposite and closer to the
responses induced by the other predators. U tadpoles
metamorphosed earlier, but at a similar size to G tad-
poles. There was no evidence that growth rate was
Communicated by Roland Brandl
A. Laurila (&) Æ S. Pakkasmaa
Population Biology/ Department of Ecology and Evolution,
Evolutionary Biology Center, Uppsala University,
Norbyvägen 18d, 75236 Uppsala, Sweden
E-mail: Anssi.Laurila@ebc.uu.se
Fax: +46-18-4716424
Present address: S. Pakkasmaa
National Board of Fisheries, Institute of Freshwater Research,
17893 Drottningholm, Sweden
J. Merilä
Ecological Genetics Research Unit, Department of Bio- and
Environmental Sciences, University of Helsinki, P.O. Box 65,
00014 Helsinki, Finland
affected by predator treatments, but tadpoles meta-
morphosed later and at larger size in the predator
treatments. G tadpoles survived better in the presence of
free-ranging predators than U tadpoles. These results
suggest that in these two populations, low growth rate
was linked with low activity and increased hiding,
whereas high growth rate was linked with high activity
and less hiding. The differences in behaviour may
explain the difference in survival between the popula-
tions, but other mechanisms (i.e. differences in swim-
ming speed) may also be involved. There appears to be
considerable differentiation in antipredator responses
between these two R. arvalis populations, as well as with
respect to different predators.
Keywords Growth rate Æ Inducible defences Æ
Plasticity Æ Population differentiation Æ Predation
Introduction
Many organisms exhibit adaptive intraspecific differen-
tiation in growth and development rates, which is often
related to large-scale climatic variation (Endler 1977;
Conover and Schultz 1995; Arendt 1997). While these
differences appear common in nature, their interplay
with other important fitness-related traits remains ob-
scure (Arendt 1997; Blanckenhorn 2000; Mangel and
Stamps 2001; Angilletta et al. 2003). For example, as
high growth rate is generally considered to increase fit-
ness, the occurrence of submaximal growth rates in
many populations is perplexing and suggests that high
growth rates must incur costs in other fitness compo-
nents. However, although several hypotheses have been
put forward to explain these costs, they are not well
understood (Arendt 1997).
One important cost of high growth rate may be in-
creased predation risk. Predator–prey interactions often
involve plastic modifications in prey behaviour,
morphology, and life history (Lima 1998a, b; Tollrian
and Harvell 1999). Evolution of differences in growth
586
and development rates may be accompanied by changes
in antipredator defences and, consequently, vulnerability
to predation (Sih 1987). Indeed, there is evidence that
individuals with high growth rates are more vulnerable
to predation, and this seems to be connected with dif-
ferentiation in traits fundamentally associated with
predation avoidance (activity and swimming speed;
Gotthard 2000; Billerbeck et al. 2001; Lankford et al.
2001; Munch and Conover 2003).
If antipredator defences are costly, environmental
heterogeneity in predator regimes is expected to cause
differentiation in these defences. While the plastic de-
fences are likely to have evolved in response to spatial
and temporal variation in predation risk, geographical
variation in these defences is not well known. Several
studies have reported population differentiation in
behavioural or morphological antipredator responses
(e.g. Riechert and Hedrick 1990; Spitze 1992; De Me-
ester 1993; Huntingford et al. 1994; Magurran 1998;
Storfer and Sih 1998; Lardner 1998; Relyea 2002).
However, these studies do not usually consider how
antipredator responses are affected by variation in other
fitness-related traits, and there are few studies exploring
the link between inducible defences and adaptive varia-
tion in other traits over environmental gradients
(Lardner 1998; Relyea 2002).
Another source of variation in antipredator defences
is the structure of predator communities. Predator
communities typically consist of multiple species, and as
predators differ in their hunting strategies, natural
selection should favour different defences against dif-
ferent predators (Sih et al. 1998). There are several
examples that prey animals can differentiate between
different predators and respond in an adaptive manner
(e.g. Tollrian and Dodson 1999; DeWitt et al. 2000;
Relyea 2001, 2003; Van Buskirk 2001; Jones et al. 2003;
Teplitsky et al. 2005), but only few studies have com-
pared several populations to several different predators
(Tollrian and Dodson 1999). As predator communities
differ, so also may the strength of selection imposed by
different predators, opening a possibility to quantitative
or qualitative variation in the expression of inducible
defences. Such variation is likely to be stronger with
predators that show strong spatial variation in their
distribution.
Many larval amphibians have well-developed anti-
predator defences in terms of inducible behaviour (re-
duced activity and spatial avoidance) and morphology
(changes in shape of tail and body; McCollum and Van
Buskirk 1996; Lardner 1998; Relyea 2001; Van Buskirk
2002). In this study, we compared larval growth and
development rates, as well as behavioural and morpho-
logical defences in two geographically separated popu-
lations of the moor frog Rana arvalis in outdoor tanks in
the presence and absence of three different predators. In
addition, we compared survival of the tadpoles in the
presence of free-ranging predators in a separate labo-
ratory experiment. One of the populations originates
from the Baltic island of Gotland (hereafter G) and the
other from Uppland (hereafter U) on the Swedish
mainland. We have previously found that these popu-
lations differ in growth and development rates, G
showing slower growth and development rates than U
(Laurila et al. 2002; P. -A. Crochet et al., unpublished).
Two of the predators we used, Aeshna dragonfly larvae
and smooth newts Triturus vulgaris, are common in both
areas. The third predator, threespine stickleback Gast-
erosteus aculeatus is a common littoral fish in the Baltic
Sea. Sticklebacks are common in G ponds which are
close to the coast and often connected to the sea by small
streams. However, they are absent in U ponds which are
situated far (ca. 80 km) from the coast and are not
connected to the sea. The predators differ in their
hunting mode: dragonfly larvae are mainly ambush
predators, sticklebacks pursue tadpole prey, whereas
newts are active predators, but do not usually pursue the
tadpoles after an unsuccessful first attack. While all
these predators prey most effectively on small tadpoles,
sticklebacks and newts are more strongly gape-limited
than dragonflies.
We addressed four main questions. First, are the
population differences in growth and development rates
reflected in the antipredator defences of the tadpoles?
Specifically, do the fast-growing U tadpoles show
weaker antipredator defences than the slow growing G
individuals. Second, do the populations differ in survival
when exposed to free-ranging predators? If high growth
rate is associated with weaker antipredator behaviour,
we expect U to have higher mortality than G. Third, do
the tadpoles show different responses against different
predators? We expect to find the strongest differences
between the responses to dragonfly and stickleback as
the difference in the hunting mode is largest between
these predators. Finally, do the populations differ in
their responses to specific predators? We mainly ex-
pected to find a difference in response to stickleback, as
it only occurs in G ponds.
Methods
Rana arvalis is a relatively small brown frog that has a
wide palearctic distribution from Western Europe to
Eastern Siberia (Gasc et al. 1997). It breeds in a wide
range of freshwater habitats, but avoids the most
ephemeral ponds. At the onset of the breeding season in
April, we collected adult R. arvalis from breeding sites in
Valar, Gotland (5700¢ N, 1813¢ E), and in Stora Almby
(5952¢ N, 1724¢ E) in the province of Uppland on the
Swedish mainland. The tadpoles originated from crosses
performed in the laboratory following Laugen et al.
(2003). For both populations, five pairwise crosses were
performed on the same day producing five full-sib
families for both populations. The use of artificial
crosses ensured that the tadpoles were of equal age and
had no prior experience with predators. The eggs from
each family were divided into six 1 l containers and
reared in room temperature in laboratory until hatching.

After hatching, the tadpoles were pooled into two pop-
ulation-specific containers and 600 seemingly healthy
tadpoles from both populations were randomly taken in
the experiment.
Late-instar larvae of the dragonfly Aeshna cyanea and
adult smooth newts were captured in ponds around
Uppsala. Adult threespine sticklebacks were captured in
the littoral zone of the Baltic in southern Gotland. Be-
fore the experiment, the sticklebacks were gradually
habituated to freshwater in the laboratory, and no fish
losses occurred during that time. When not in the
experiment, the sticklebacks were kept in circular plastic
pools (diameter 150 cm, depth 30 cm) outdoors and fed
R. arvalis and R. temporaria tadpoles and aquatic
invertebrates daily. The newts were kept outdoors in
covered plastic tanks filled with 20 l of well water and
fed tadpoles ad libitum. The dragonflies were kept in
individual 0.25 l plastic vials outdoors and fed two
tadpoles every third day.
Experimental design
The experiment was conducted in a field at Segelbo,
Heby municipality, 50 km northwest from Uppsala. 40
large plastic containers (80·40·35 cm), arranged in five
spatial blocks, were used as experimental units. On April
11, the containers were filled with 80 l of well water. In
addition, 2 l of pond water were added to each container
to provide an inoculum of phyto- and zooplankton.
Each container received 6 g of rabbit pellets and 10 g of
dried aspen (Populus) and birch (Betula) leaves to pro-
vide nutrients for the system. Leaves also acted as
structural complexity and provided shelter for the tad-
poles, which was further increased by adding four 8 cm
pieces of plastic pipe (diameter 2.5 cm) in each con-
tainer. The containers were covered with plastic mos-
quito net to prevent insects from colonising the
containers. A cylindrical predator cage (diameter 11 cm,
height 21 cm) made of transparent plastic film with a
double net bottom (mesh size 1.5 mm) was hung 7 cm
above the tank floor in each experimental unit.
The experiment was initiated on April 29 (day 0 of
the experiment), when tadpoles were at developmental
stage 25 (Gosner 1960). 30 tadpoles from either U or G
population were released in to each container. The
predators were introduced into the cages on day 1. Each
container received no predator (control; empty cage) or
one stickleback, newt, or dragonfly larva.
An additional 4 g of rabbit pellets were added to each
container on day 22. Otherwise, the tadpoles relied on
tree leaves and periphyton as food. The predators were
fed ca. 300 mg of R. arvalis tadpoles every second day.
The present experimental design cannot differentiate
whether the tadpoles respond to visual or chemical (i.e.
kairomones) cues from the predator, or to chemical
alarm cues released by the captured prey (e.g. Laurila
et al. 1998; Schoeppner and Relyea 2005). However, any
difference in the response variables between the three
587
predator treatments would indicate that the tadpoles are
reacting to the predators rather than to cues from preyed
conspecifics.
Response variables
Tadpole behaviour was recorded twice per day (between
09 and 12 a m) during day 3–6 and 27–32. The con-
tainers were approached carefully and the numbers of
active (judged by moving tail) and visible tadpoles were
scored.
Eight tadpoles were sampled from each container on
days 13 and 33 and stored in 10% formalin for mea-
surements. Body length (from the tip of the nose to the
end of the body wall), maximum body depth, tail length
(from the body terminus to the tip of the tail), maximum
tail-muscle depth (at the base of the tail), and maximum
tail depth were later measured with a digital calliper to
the nearest 0.01 mm. The same person conducted the
measurements blindly with respect to the treatments.
When the tadpoles were approaching metamorphosis
(emergence of at least one forelimb; Gosner 1960) the
containers were checked daily. Metamorphosed tadpoles
were removed from the containers, blotted dry, weighed
to the nearest 1 mg with an electronic balance and the
length of larval period (from the start of the experiment)
was determined for each individual.
Statistical analyses
We used a 2·4 randomised block design with a combi-
nation of four predator treatments and two population
origins replicated over five spatial blocks. All the vari-
ables were considered as fixed. Behavioural and larval
life history data were analysed with MANOVAs fol-
lowed by univariate ANOVAs using type III sums of
squares. Due to the change in density and composition
of individual tadpoles between the sampling periods we
preferred to use a normal MANOVA instead of a re-
peated-measures MANOVA in the behavioural analy-
ses. Since values of morphological traits are dependent
on general body size, tadpole morphology was analysed
with principal component analyses (PCA) with correla-
tion matrices to reduce variation in initial morphological
measurements into composite variables of size and
shape. In all analyses, all traits loaded strongly and
positively on the first axis, revealing this axis to be a size
component. To obtain size-corrected measurements of
the morphometric traits, we regressed the traits against
PC1 scores and analysed the residuals (shearing method;
Bookstein 1991). PC1s from the analyses were used as an
index of larval growth. Finally, we performed MANO-
VAs on all the morphological variables followed by
univariate ANOVAs. The analyses were conducted with
Systat 10 statistical package (Wilkinson 2000). Tadpole
mortality was analysed with type III general linear
models with a logit link function and binomial error
588
structure as implemented in the GENMOD procedure
of SAS (Allison 1999).
Analyses on behavioural variables were conducted on
arcsin-squareroot transformed container-specific pro-
portions, whereas the ANOVAs on morphology and life
history traits were performed on container means. One
experimental unit (U population, newt treatment) suf-
fered sudden heavy mortality around day 20 and was
excluded from all subsequent analyses.
Survival in the presence of free-ranging predators
Five fresh spawn clumps of R. arvalis were collected in
April at the same two localities as in the previous
experiment and brought into laboratory. The egg clumps
were allowed to develop until Gosner stage 25 in family-
specific 3 l vials in room temperature. Thereafter, the
tadpoles from each population were pooled and 90
tadpoles were randomly allotted to each of the plastic
tanks (80·40·35 cm3) filled with 70 l of aged tap water
(day 0). For both populations we used six tanks, two of
which were control (predator-free) tanks, two had a
caged stickleback in a similar setting as in the previous
experiment, and two had a caged dragonfly larva. The
tadpoles were raised indoors under constant (16L: 8D)
photoperiod in room temperature (mean water temper-
ature during the experiment 19.025±0.089 (SE) C,
n=23 days) and fed slightly boiled spinach ad libitum.
The predators were fed 300 mg of R. arvalis tadpoles
every second day.
The trials with free-ranging predators were conducted
between days 18 and 21. Due to differences in breeding
phenology, the tests with G tadpoles were started eigh-
teen days before those with U tadpoles. For both pop-
ulations, naı̈ve and induced tadpoles were tested both in
the presence of a free ranging stickleback and in the
presence of a dragonfly larva. Induced tadpoles were
tested only with the predator species they had been
raised with. The experimental arenas were plastic con-
tainers (49·26·25 cm3) filled with aged tap water to the
depth of 10 cm. Ca. 40 dried aspen leaves were added in
each container to provide structural complexity. The
containers were arranged in a single row on a shelf in a
laboratory room illuminated with natural light.
At the start of each experimental run, ten randomly
selected tadpoles from one population and predator
treatment were added to each container. After one hour,
one predator (a stickleback or a dragon fly larva) was
added to each container. As survival of R. arvalis tad-
poles is very good under laboratory conditions, we did
not conduct control trials without predators. The trial
was terminated after 23 h, the number of surviving
tadpoles was recorded, and the total length of each
predator was measured. We also carefully examined all
the tadpoles for injuries, which are indicative of predator
hunting activity even in the absence of killed tadpoles
(Morin 1985). Each predator and prey were used only
once in the experiment. Trials ranging between 8 and 10
were conducted for each treatment combination. The
total length (from tip of the snout to the tip of tail) of ten
randomly selected tadpoles from each container was
measured before the start of the experiments: There was
a nonsignificant tendency for the U tadpoles [mean
length 29.32±0.50 (SE)] to be larger than G tadpoles
(mean length 28.16±0.29; t10= 1.98, P=0.076).
We analysed tadpole survival with generalised linear
models in Proc GENMOD in SAS. Population origin,
predator type and induction treatment were used as
fixed factors and predator length as a covariate. The
three-way interaction among the factors was not signif-
icant and was removed from the final model.
Results
Behaviour
MANOVA showed significant effects of population
(Wilk’s k=0.176, F4,24=28.06, P<0.001), predator
(Wilk’s k=0.063, F12,63=9.80, P<0.001) and popula-
tion · predator interaction (Wilk’s k=0.259, F12,63=
3.54, P<0.001). Examination of univariate ANOVAs
indicated that early in the experiment (days 3–6) both
population and predator treatments had significant
effects on the numbers of tadpoles visible (Table 1,
Fig. 1). In general, tadpoles were hiding more in newt
and stickleback treatments, whereas there was no differ-
ence between control and dragonfly treatments. How-
ever, the difference between treatments was much larger
in G than in U tadpoles explaining the significant pop-
ulation · predator interaction (Table 1, Fig. 1). Predator
treatments had a strong effect on activity in both popu-
lations with tadpoles in the predator treatments being less
active than those in control (Table 1, Fig. 1).
During the latter observation period (days 27–32) U
tadpoles were more often visible than G tadpoles
(Table 1, Fig. 1). While the predator main effect was not
significant, there was highly significant popula-
tion · predator interaction (Table 1). This was due to
the fact that G tadpoles were most visible in stickleback
treatment, whereas in U, the opposite was true (Fig. 1).
In both populations, tadpoles tended to hide somewhat
more in newt and dragonfly treatments than in the
control treatment, but this difference was not significant
(Table 1, Fig. 1).
U tadpoles were much more active than G tadpoles
during the latter observation period (Table 1, Fig. 1).
Table 1 Univariate ANOVA F-values for behaviour of R. arvalis
tadpoles during days 3–7 (P1) and 27–31 (P2)
Source df Activity P1 Visible P1 Activity P2 Visible P2
Population 1,27 1.58 11.48*** 69.67**** 82.71****
Predator 3,27 19.72**** 24.35**** 6.49*** 2.35*
Pop. · Pred. 3,27 1.76 4.1** 7.34*** 12.84****
Block 4,27 0.95 1.71 1.16 1.07
*P<0.10; **P<0.05, ***P<0.01; ****P<0.001
 589

Fig. 1 Proportion of tadpoles Control Stickleback

visible and active in the two
Newt Dragonfly
populations and four predator
treatments. The values 1 0.3
represent least square
means ± SE

Proportion visible

Proportion active
0.75
0.2

0.5

0.1
0.25

0 0
Gotland Uppland Gotland Uppland Gotland Uppland Gotland Uppland

Days 3-6 Days 27-32 Days 3-6 Days 27-32

Predator treatments reduced activity in dragonfly and
newt treatments (Table 1, Fig. 1). U tadpoles decreased
activity also in the presence of sticklebacks, whereas in
G activity did not differ from the control, causing the
significant population · predator interaction. In gen-
eral, U tadpoles were more active in the presence of
predators than G tadpoles were in control treatment
(Fig. 1).
Morphology
MANOVA showed no significant effects of population
origin on morphology on day 13 (Wilk’s k=0.721,
F5,24=1.86, P=0.14). However, MANOVA was close to
significance on day 32 (Wilk’s k=0.649, F5,23=2.49,
P=0.061), and univariate ANOVAs indicated that G
tadpoles tended to have relatively deeper tail than U
tadpoles (Table 2, Fig. 2).
MANOVAs indicated significant effects of predator
treatments on tadpole morphology on day 13 (Wilk’s
k=0.066, F15,66=7.45, P<0.001) and on day 32 (Wilk’s
k=0.116, F15,63=5.03, P<0.001). In general, tadpoles
had shorter body and deeper tail in the predator treat-
ments (Table 2, Fig. 2). There was some heterogeneity in
response to different types of predators on day 13: the
response in tail depth was weaker in stickleback treat-
ment, and body was shallower in newt treatment (Ta-
ble 2, Fig. 2). In addition, tail muscle was shallower in
stickleback treatment in G, whereas it was deeper in the
presence of newt in U causing a significant popula-
tion · predator interaction. However, MANOVA found
no significant population · predator interaction in tad-
pole morphology on day 13 (Wilk’s k=0.449,
F15,66=1.49, P=0.133).
Also on day 32, the strongest response was in tail
depth with strongest effect in dragonfly treatment,
somewhat weaker in the presence of newt and only a
weak response in stickleback treatment (Table 2, Fig. 2).
In other traits, the interpretation of predator effects was
complicated by population · predator interactions,
which were also indicated by MANOVA (Wilk’s
k=0.408, F15,63=1.64, P=0.089). G tadpoles had rela-
tively longer and deeper body in the presence of stick-
leback, whereas body shape was not affected in other
predator treatments (Table 2, Fig. 2). U tadpoles had
shorter body in stickleback and dragonfly treatments
and shallower body in all predator treatments as com-
pared to control. Presence of stickleback also induced a
decrease in tail length in G, and tail muscle tended to be
shallowest in stickleback treatment in G but deepest in
U (Table 2, Fig. 2).
Larval survival and life history
The proportion of G tadpoles surviving until meta-
morphosis (0.925±0.027) was significantly higher than

Table 2 Univariate ANOVA

F-values for morphology Source df Body length Body depth Tail length Tail depth Tail muscle depth
(residual values, see text) of
R. arvalis tadpoles on days 13 Day 13
and 33 Population 1,28 0.19 0.85 0.03 5.44* 0.46
Predator 3,28 8.79**** 3.01** 2.53* 40.03**** 3.24**
Pop. · Pred. 3,28 0.77 0.99 0.08 2.22 3.27**
Block 4,28 1.02 2.14 1.30 2.64* 2.83**
Day 33
Population 1,27 0.51 0.89 0.99 7.66*** 1.29
Predator 3,27 3.12** 2.01 4.42** 18.86**** 0.02
Pop. · Pred. 3,27 4.48** 3.15** 3.26** 0.54 3.08**
*P<0.10; **P<0.05; Block 4,27 3.43** 2.66* 1.64 2.03 2.05
***P<0.01; ****P<0.001
590

Fig. 2 Body shape (residual 0.75 0.3

body length, body depth, tail
length, tail depth and tail
0.5 0.2
muscle depth) of tadpoles on

Residual body length

Residual body depth

day 13 and 32 in the two
populations and four predator 0.25 0.1
treatments. The values
represent least square 0 0
means ± SE
-0.25 -0.1

-0.5 -0.2

-0.75 -0.3

0.8 0.4

Residual tail length 0.6

0.4 0.2

Residual tail depth

0.2
0 0
-0.2
-0.4 -0.2
-0.6
-0.8 -0.4
Gotland Uppland Gotland Uppland
0.15 Day 13 Day 32
Residual tail muscle depth

0.1
Control Stickleback
0.05 Newt Dragonfly

-0.05

-0.1

-0.15
Gotland Uppland Gotland Uppland
Day 13 Day 32

in U population (0.845±0.028; v21,27=4.27, P=0.039). Survival under predation risk

Predator treatments did not influence tadpole survival
(P>0.8).
MANOVA indicated significant population (Wilk’s
k=0.063, F4,24=88.94, P<0.001) and predator effects
(Wilk’s k=0.364, F12,63=2.48, P=0.01) in growth and
development, whereas population · predator interac-
tion was not significant (P=0.335). Larval growth
(indicated as PC1 scores) was significantly faster in U
than in G population, and U tadpoles metamorphosed
earlier than G tadpoles (Table 3, Fig. 3). However, there
was no difference between the populations in metamor-
phic mass. Tadpoles in the predator treatments meta-
morphosed later and at larger mass than control
individuals (Table 3, Fig. 3). The block had significant
effects on larval growth and developmental rates (Ta-
ble 3), possibly because of temperature-related envi-
ronmental variation among the blocks.
In seven of the total of 72 trials, the predator did not
show any hunting behaviour (i.e., all the tadpoles sur-
vived unharmed, always in the stickleback treatments)
and these trials were excluded from the analyses. In an
additional ten stickleback trials, all the tadpoles survived
but one or more of the tadpoles showed signs of stick-
leback hunting activity (usually tail injury). These trials
were included in the analysis. Hence the final sample size
was 65 (Fig. 4). G tadpoles survived better than U
individuals in the presence of free-ranging predators
(v2=5.44, df=1, P=0.019, Fig. 4). Dragonfly larvae
caused more mortality than sticklebacks (v2=23.83,
df=1, P<0.001, Fig. 4), and induced tadpoles raised in
the presence of predators had higher survival than naı̈ve
tadpoles (v2=4.51, df=1, P=0.034, Fig. 4). None of the
interactions was significant (P>0.24), but increasing
 591

Table 3 Univariate ANOVA

F-values (produced by Source df PC1 at day 13 PC1 at day 32 Mass at Age at
MANOVA) for larval life- metamorphosis metamorphosis
history traits of R. arvalis
tadpoles Population 1,27 124.91**** 122.85**** 0.76 113.54****
Predator 3,27 1.22 0.96 3.65** 4.06**
Pop. · Pred. 3,27 0.95 0.67 0.75 1.86
**P<0.05, ***P<0.01, Block 4,27 8.34**** 6.91**** 2.22 2.85**
****P<0.001

Fig. 3 Tadpole body size (PC1) Control Stickleback

on day 13 and 32, mass at Newt Dragonfly
metamorphosis and length of
larval period in the two 1 1
populations and four predator
treatments. The values
represent least square 0.5 0.5
means ± SE PC1 at day 13

PC1 at day 32
0 0

-0.5 -0.5

-1 -1
0.6 65
Mass at metamorphosis (g)

Larval period (d)

0.5 60

0.4 55

0.3 50
Gotland Uppland Gotland Uppland

predator size increased tadpole mortality (v2=5.44,
df=1, P=0.019).
Discussion
We found significant differentiation between the two R.
arvalis populations in growth and development rates,
behaviour, morphological defences and survival under
predation risk. Both populations showed induced re-
sponses to predators, which varied during ontogeny and
in response to specific predators. In the following, we
will discuss these findings in more detail.
Growth rate, antipredator behaviour and survival
As growth rate is considered to be ultimately linked with
individual fitness, the occurrence of submaximal growth
rates is intriguing, and several hypotheses on the costs of
high growth rates have been put forward (Arendt 1997).
Empirical studies testing the hypotheses are few, but
include examples showing that fast growing individuals
may have lower starvation endurance (Gotthard et al.
1994), competitive ability (James and Partridge 1998),
pathogen resistance (Smoker 1986; Brommer 2003) or
rate of skeletal ossification (Arendt and Wilson 2000).
We found that irrespective of predator treatment, the
fast-growing U population had higher mortality than
the slow-growing G population, possibly suggesting that
high growth rate per se may include a mortality cost.
Any of the mechanisms mentioned above may be func-
tioning, although we did not see any signs of disease or
starvation in the tadpole, and the difference could also
be due to some unknown factor.
U tadpoles had significantly higher mortality in the
presence of free-ranging predators, suggesting the
intriguing possibility that high predation risk may be a
cost of high growth rate in R. arvalis. Few other studies
have measured the trade-off between high growth rate
592
Non-induced Induced
1 6 8 studies on both tadpole and predator behaviour are
5 9
Proportion surviving
0.9
9 10
0.8
8 10
0.7
0.6
0.5
Dragonfly Stickleback Dragonfly Stickleback
Uppland
Gotland
Fig. 4 Survival of R. arvalis tadpoles in the two populations in the
presence of a free-ranging dragonfly larva or stickleback. The
values represent means ± SE. Sample sizes are indicated on the
top of the bars
and predation risk. Gotthard (2000) showed that
experimentally time-constrained, fast-growing Pararge
butterfly larvae suffered higher mortality due to preda-
tion. In the Atlantic silverside Menidia menidia, the fast
growing northern population had higher mortality than
the slow growing southern population when exposed to
predatory fish (Billerbeck et al. 2001; Lankford et al.
2001; Munch and Conover 2003). It is clear that our
study based on two populations cannot provide a
definitive answer on the question how growth rate af-
fects predation mortality. However, in accordance with
Menidia studies, we found that a fast-growing high lat-
itude population had a higher predation risk, possibly
suggesting that increased predation risk may be a com-
mon character of populations with high individual
growth rates.
We are only beginning to understand the mechanisms
behind the higher vulnerability of fast growing individ-
uals. High activity rate is likely to increase predation risk
(e.g. Sih 1987), and if high intake rate is linked to high
growth rate, active foraging may render the fast-growing
individuals more vulnerable. The relationship between
activity level and predation risk is especially well estab-
lished in amphibian larvae (e.g. Skelly 1994; Anholt and
Werner 1995), and activity and growth appear to be
positively genetically correlated in anurans (Relyea
2005). We found that U tadpoles maintained higher
activity rates even in the presence of predators, and
it seems possible that increased predation risk in
U tadpoles is mediated by their high activity level.
However, other mechanisms may also be involved in the
higher vulnerability of U tadpoles. In several fish species
(Kolok and Oris 1995; Gregory and Wood 1998;
Billerbeck et al. 2001), as well as in tadpoles (Arendt
2003), fast-growing individuals have relatively lower
sprint-swimming speed, which increases their vulnera-
bility to predation (Lankford et al. 2001). On the other
hand, tadpoles sprint speed generally increases with
body size (Van Buskirk and McCollum 2000; Arendt
2003; Teplitsky et al. 2005), which is likely to decrease
predation risk for fast-growing U individuals. Detailed
needed to disentangle the mechanisms of higher mor-
tality in U tadpoles.
The reason for the difference in growth and devel-
opment rates between the populations is currently un-
clear, and many factors can potentially affect these traits
(see Arendt 1997 for a review). Ponds in both areas
harbour diverse and dense populations of predators (A.
Laurila et al., personal observation), rendering it un-
likely that large differences in predation pressure would
occur between the populations. A more likely explana-
tion is selection imposed by climate, especially season
length and/or temperature. For example, length of
growth season is about 219 days in G and 184 days in U
(Odin et al. 1983) indicating that growth and develop-
ment in U population are more time-constrained. We
found low growth and development rates and strong
antipredator behaviour in the southern population (G),
whereas the northern (U) population had high growth
and development rates and higher activity, suggesting
that variation in seasonal time constraints may be a
driving force behind population-level variation. While
only studies using more populations can definitely re-
solve the role of climate in population differentiation,
our view is supported by the fact that increasing growth
and development rates towards higher latitudes have
been found in other Rana species living along latitudinal
gradients including Rana temporaria in Scandinavia (e.g.
Berven and Gill 1983; Laugen et al. 2003; Lindgren and
Laurila 2005). Similarly, studies on insects have found
that larval growth and development are faster in U than
in G, further supporting the role of climate in creating
this variation (Nylin et al. 1996).
Variation in antipredator responses
In line with some earlier studies (Arnqvist and Johans-
son 1998; Van Buskirk 2001, 2002), we found consid-
erable ontogenetic variation in the antipredator
defences. Furthermore, the responses differed between
the populations suggesting a role for local selection
pressures in shaping them. The most striking difference
between the populations in behaviour was increased
hiding and low activity level in G late in the experiment.
We suggest that the relatively high early activity levels
found in the absence of predators in both populations
may reflect the fact that rapid growth is especially
important for small tadpoles in order to gain a headstart
in competition, or to avoid gape-limited predators. After
passing the most vulnerable early stages, less time lim-
ited G may afford the lower activity level and slower
growth and development rates, whereas U increases
activity further as development proceeds. Another po-
tential explanation for the difference in activity between
the populations is that mortality of older tadpoles due to
predation is invariably higher in G than in U popula-
tion, which might have selected for low activity levels.

Several studies have shown that prey animals can
dynamically balance their behaviour during ontogeny to
minimize the risk of being eaten. For example, Van
Buskirk (2001) showed that only small R. temporaria
tadpoles avoided Triturus alpestris newts, which are
strongly gape-limited and can only feed effectively on
small tadpoles.
R. arvalis tadpoles showed differential behavioural
responses to different predators suggesting that they
were able to differentiate between the predators. For
example, young tadpoles showed reduced activity in
response to all three predators, whereas they showed
increased hiding behaviour only in response to newt
and stickleback. Both newts and stickleback are active
predators moving in the water column, and hiding in
pond bottom is probably an efficient antipredator
tactic against them. On the contrary, dragonfly larvae
are hiding sit-and-wait predators. Young tadpoles in
dragonfly treatment avoided hiding in tank bottom
and instead stayed immobile on tank walls. However,
later in the experiment, such behaviour was not evi-
dent suggesting that the tadpoles relied more on
inactivity and morphological defences later in their
development.
The responses in induced morphology to the two
common predators were broadly similar in the two
populations and in accordance with previous studies on
this and other amphibian species (increased relative tail
depth and decreased body size; e.g. McCollum and Van
Buskirk 1996; Relyea 2001; Van Buskirk 2001). How-
ever, the responses of older tadpoles to sticklebacks
differed between the populations. G tadpoles had rela-
tively longer and deeper bodies, shorter tail, shallower
tail muscle and slightly deeper tail fin in the presence of
stickleback, whereas the response in U was more similar
to the dragonfly treatment except that tail muscle tended
to be deeper in the presence of stickleback, and tail fin
depth was not different from control. These differences
may reflect adaptation to stickleback predation as G
tadpoles commonly co-occur with sticklebacks, but U
tadpoles do not. However, despite the differences in
morphological defences, the population difference in
survival was not dependent on the predator type, and
survival was generally improved by the induction treat-
ment in both populations.
While several studies have studied morphological
plasticity of anuran larvae in response to insect preda-
tors, morphological responses to fish have received less
attention (but see Relyea 2001; Teplitsky et al. 2003). In
R. dalmatina, a closely related species to R. arvalis, the
morphological responses induced by sticklebacks in-
cluded deeper tail muscle and longer tail fin. Deeper tail
muscle increased swimming speed and the plastic chan-
ges in R. dalmatina were to the same direction as selec-
tion on morphology by free-ranging sticklebacks
(Teplitsky et al. 2003, 2005). Sticklebacks are gape-lim-
ited and can catch large tadpoles only by repeated
attacks (A. Laurila, C. Teplitsky, personal observation).
The increased relative body length and depth may
593
protect the G tadpoles from stickleback attacks and give
them enough time to find a shelter (see also Kishida and
Nishimura 2004). Intriguingly, body form in G tadpoles
did not change in response to dragonflies and newts
possibly reflecting the differences in the hunting strate-
gies. On the other hand, the increase in tail muscle in U
is similar to the changes in R. dalmatina, which were
demonstrated to be of survival value under stickleback
predation (Teplitsky et al. 2005). In a comparison of six
tadpole species, Relyea (2001) found that the responses
induced by the mudminnow (Umbra limi) varied widely
among the species and the traits. The variation among
species and populations may suggest that there are sev-
eral potential morphological responses to fish predators.
Tadpoles in the predator treatments metamorphosed
later but at a larger size than those from the control
treatment. There was no evidence that tadpole growth
rate was influenced by predator treatments; instead
tadpole development was negatively affected by predator
presence. Similar results have been obtained in several
studies of ranid frogs (e.g. Laurila et al. 1998; Altwegg
2002; Van Buskirk and Saxer 2002). Late metamor-
phosis may have a negative influence on later fitness
(Smith 1987; Berven 1990). However, the possible neg-
ative effects of later metamorphosis may be largely
compensated by the larger body mass at metamorphosis.
Indeed, Altwegg and Reyer (2003) found that the posi-
tive effects of increased metamorphic size on terrestrial
survival of juvenile pool frogs (Rana lessonae) were al-
ways stronger than the negative effects of late meta-
morphosis. If similar relationship holds true for R.
arvalis, the direct fitness costs of antipredator defences
are likely to be small.
In conclusion, we found considerable differentiation
in antipredator defences between two R. arvalis popu-
lations differing in growth and development rates. The
slow-growing population with stronger antipredator
behaviour also exhibited significantly higher survival in
the presence of free-ranging predators. Our results sug-
gest that variation in activity levels and behavioural
defences may have an important role in mediating life
history differentiation among populations. However, the
mechanisms of differential predation mortality need to
be investigated further.
Acknowledgments We thank Pierre-André Crochet, Katja Enberg,
Maria Järvi-Laturi, Beatrice Lindgren, Outi Pihlajamäki and Katja
Räsänen for field and laboratory assistance, and Nina Peuhkuri,
Miguel Tejedo, Celine Teplitsky and an anonymous reviewer for
helpful comments on the manuscript. The study was performed
with the permission C 72/1 from Uppsala county ethical committee
for animal experiments. Our research was funded by the Swedish
Research Council (AL) and the Academy of Finland (SP, JM).
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