Current Zoology

59 (3): 326334, 2013

Memory of conspecifics in male salamanders Plethodon

cinereus: Implications for territorial defense
Nancy R. KOHN1*, Jennifer M. DEITLOFF2,3, Schuyler F. DARTEZ1,4, Michelle M.
WILCOX1, Robert G. JAEGER1,5
1

Department of Biology, University of Louisiana at Lafayette, Lafayette, LA 70504, USA

Department of Ecology, Evolution, and Organismal Biology, Iowa State University, Ames, IA 50011, USA

Abstract We investigated how exposure duration (time that two individuals initially interact) and separation interval (time between the initial interaction and a subsequent interaction) affect recognition memory of conspecifics in male red-backed salamanders Plethodon cinereus. Previous studies have demonstrated that this species aggressively defends territories. We recorded
aggressive behavior to assess recognition memory, because aggression is more intense toward previously unencountered individuals compared to previously encountered individuals in this species. We found that with 15-min exposures and 5-day separation intervals, focal males did not spend significantly more time threatening unfamiliar intruders than familiar intruders. After
either 8-hour exposures and 5-day separation intervals and 5-day exposures and 5-day separation intervals, focal males spent significantly more time threatening unfamiliar intruders than familiar intruders. These results suggest that male red-backed salamanders can remember familiar conspecifics (e.g., territorial neighbors) after at least an 8-hour exposure duration and that memory
persists at least as long as 5 days. After 5-day exposure and 15-day separation intervals, we found no significant difference in aggressive behavior toward familiar and unfamiliar intruders. Long separation intervals (15 days) may lead either to loss of memory
of previously familiar individuals or, alternatively, aggressive reassessment of individuals as only a change in behavior indicates
positively that memory has occurred. Thus, variance in territorial defense within an individual may depend on its ability to recognize conspecific males [Current Zoology 58 (3): 326334, 2013].
Keywords

Individual recognition, Memory, Plethodon cinereus, Red-backed salamanders, Territoriality

Many species of animals engage in social interactions

that require some form of memory (Shettleworth, 1998;
Dukas, 2004), and memory differs among species according to the ecological requirements of each species
(Shettleworth, 2001; Odling-Smee et al., 2008). More
specifically, memory can be important for avoiding
predators, searching for food or mates, navigation, and
engaging in various types of social interactions. Climate-related environmental conditions may also impact
memory, as memory may be more important for survival in harsher environments than less harsh environments (Freas et al., 2012).
Memory involves the capacity to store, retain, and
retrieve information from the past and is studied by
looking for a change in behavior due to experience
(Dukas, 1998; Shettleworth, 1998; Kawecki, 2010).
While a change in behavior indicates memory, a lack of
response or no change in behavior does not necessarily

mean that an animal has forgotten. A lack of response

could be because an individual has forgotten, or the individual remembers but there is no functional reason to
change its behavior. We studied memory in the context
of recognizing familiar individuals during territorial
defense. Following Ryan and Rand (1993) and Alberici
de Barbiano et al., (2012), we define recognition as a
behavioral response to a single stimulus (individual) and
discrimination as when an individual responds more or
less strongly to one stimulus over another stimulus. In
order to recognize individuals one needs to be able to
assess others based upon perceptual traits expressed by
or associated with those individuals (e.g., as in kin recognition: Waldman et al., 1988). In particular, individual
recognition usually involves the association of specific
information (e.g., auditory, visual, chemical cues) with
memories of past experiences with a specific individual
(Johnston and Bullock, 2001).

Received July 31, 2012; accepted Sept. 27, 2012.

Corresponding author. Current Address: Nancy R. Kohn, Department of Biology, University of Missouri-St. Louis, One University Boulevard,
St. Louis, MO 63121, USA; E-mail: nancy.r.kohn@gmail.com
3
Current address: 110 Insectary, Pammel Drive, Iowa State University, Ames, IA 50011, USA. 4 Current address: The Louisiana Department of
Wildlife and Fisheries, White Lake Wetlands Conservation Area, Gueydan, LA 70542, USA. 5 Current address: 100 Simon Latour Road,
Carencro, LA 70520, USA
2013 Current Zoology

KOHN NR et al.: Memory of conspecifics in male salamanders

Individual recognition is most likely found in species

in which 1) the perceptual traits or signals vary at the
individual level (i.e., phenotypic variation: Alberts and
Werner, 1993; Sheehan and Tibbetts, 2010) and 2) individual conspecifics encounter each other repeatedly
(Aragon et al., 2001). Examples include permanent
multi-purpose territories (Glinski and Krekorian, 1985;
Temeles, 1994), colonies of closely interacting individuals (Myrberg and Riggio, 1985), or dominance hierarchies (McLeman et al., 2005; Dreier and DEttorre,
2009). However, individual recognition does not necessarily occur if there is phenotypic variation in a species
but not repeated social interactions or vice versa (Dreier
and DEttorre, 2009; Sheehan and Tibbetts, 2010). Individual recognition has been studied in many taxa, including mammals (Johnston and Bullock, 2001; Mateo,
2006), birds (Temeles, 1994; Botero et al., 2007), lizards
(Alberts and Werner, 1993; Carazo et al., 2008), frogs
(Bee et al., 2001; Bee and Gerhardt, 2002), fish (Myrberg and Riggio, 1985), insects (Tibbetts, 2002; Sheehan and Tibbetts, 2008; Dreier and DEttorre, 2009),
and lobsters (Gherardi et al., 2010). However, many
studies do not specifically control for how long the individuals initially have to interact and separation interval, both of which can be important for memory
(Menzel et al., 2001; Nairne, 2002; Cowan, 2008).
Red-backed salamanders Plethodon cinereus are
found under rocks and logs and in the leaf litter on forest floors of northeastern North America (Jaeger, 1980b;
Mathis, 1990, 1991). Both males and females defend
territories intersexually (Lang and Jaeger, 2000) during
the courting and non-courting seasons (Mathis, 1989).
These territories function as places to sequester food
(Jaeger, 1980a; Jaeger and Forester, 1993), as refuges
from desiccation (Jaeger, 1980a; Gabor, 1995), and as
potential courtship sites (Mathis, 1991). At our research
site in Virginia, USA, adults of both sexes advertise
their territories with pheromonal markings (Jaeger et al.,
1986; Horne and Jaeger, 1988) and defend their territories with visual displays (Jaeger, 1984) and biting (Jaeger, 1981).
Previous research suggests that individual recognition occurs in red-backed salamanders (Jaeger, 1981;
Gillette et al., 2000; Jaeger et al., 2002; Joseph et al.,
2005; Kohn and Jaeger, 2009). Red-backed salamanders
can distinguish substrate markings of familiar from unfamiliar conspecifics (McGavin, 1978), and both adult
males and females employ dear enemy recognition in
which individuals are less aggressive and more submissive toward familiar territorial neighbors than toward

327

strangers (Jaeger, 1981). In addition, familiarity leads to

decreased aggression between opposite-sex salamanders
(Joseph et al., 2005), and male red-backed salamanders
remember individuals based just on visual cues or
chemical cues after a 5-day separation in the laboratory
(Kohn and Jaeger, 2009).
In regards to memory, the longer the exposure to an
individual, the stronger the memory trace for that individual should become (Ruiz et al., 2004). Therefore, we
used different exposure durations and separation intervals to answer the following questions: 1) How long do
male salamanders have to interact before they become
familiar with each other? 2) At what length of separation time do salamanders behave towards familiar individuals as they do towards unfamiliar individuals (either
because of forgetting or the need to reassess potential
competitors)? We conducted three experiments to test
whether or not exposure duration impacts the memory
of individuals in territorial male red-backed salamanders.
We predicted that longer exposure durations would lead
to reduced aggression toward familiar individuals during second encounters. We chose a 15 min exposure
(experiment 1) to simulate an intruder merely passing
through a focal salamanders territory, an 8-hour exposure (experiment 2) to simulate an individual who has
recently set up a territory nearby, and a 5-day exposure
(experiment 3) to simulate an established territorial
neighbor (as in Jaeger et al., 1981). The above three
experiments all contained a 5-day separation period
after the initial exposure duration. In addition to experiment 3, 5-day exposure and 5-day separation, we
also examined the effect of a longer separation interval
(15-day) after the initial 5-day exposure for a total of
four experiments. We predicted that salamanders should
remember familiar individuals after a 5-day separation
interval based on the results of Kohn and Jaeger (2009).
However, we had no predictions, based on previous
research, whether or not salamanders can remember
familiar individuals after a longer 15-day separation
interval.

Materials and Methods

1.1 Collection and Maintenance of Salamanders

We collected single adult (>32 mm SVL or snoutvent length: Sayler, 1966) male red-backed salamanders
near Mountain Lake Biological Station (MLBS), Giles
County, Virginia, USA. We define single males as being
alone under a rock or log, and not those found cohabiting with other conspecifics. We used only male salamanders in these experiments in order to avoid compli-

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cations from having females in different reproductive

states (gravid vs. not gravid). We placed each salamander in a separate jar containing a paper towel moistened
with spring water for transportation (in ice chests) to our
laboratory in Lafayette, Louisiana, USA. We released
the salamanders near the site of capture following the
completion of these experiments.
In the laboratory, we placed each salamander in a
Petri dish (14 1.5 cm) containing a filter paper moistened with spring water. We fed the salamanders approximately 20 fruit flies Drosophila melanogaster per
week, and filter papers were replaced about every two
weeks until the experiments began. During experiments
1, 2, and 3, we fed the salamanders on days 1, 4, 6, and
9. During experiment 4, we fed the salamanders on days
1, 4, 6, 10, 15, and 19. The laboratory was maintained at
18 3 C with a 12:12 h light: dark cycle.
We determined the sex of each salamander by placing
it in a clear plastic bag and holding it up to a bright light
to determine the presence or absence of pigmented testes (Gillette and Peterson, 2001); females and juveniles
were released back to where we found them in the forest.
Salamanders with autotomized tails were not used in
these experiments, because red-backed salamanders
alter their aggressive interactions with others based on
their opponents tail condition (Wise and Jaeger, 1998).
We did not conduct one large, randomized, experimental design because this would have required a total
of 550 adult male red-backed salamanders to achieve a
large enough sample size, and we would have stressed
an additional 2600 individuals to obtain only single
adult males. Consequently, we divided our research into
four separate experiments and we were unable to use
statistical analyses between or among experiments.
1.2 Experiment 1: 15-min exposure and 5-day
separation interval
We collected male salamanders (n = 175) from 79
October 2004 and conducted this experiment between
January and March 2005 from 12: 0016: 00 h. We divided the salamanders into 26 groups of five. Within
each group, we randomly chose one male as the focal,
territorial salamander that would be tested for memory
of the other males that were randomly assigned as intruders. We tested the focal salamanders in each of the
three treatments: 1) familiar intruder trials, 2) unfamiliar
intruder trials, and 3) surrogate control trials, in random
order, every third week, until each focal salamander had
been through all three treatments. A surrogate consisted
of a rolled piece of paper towel (moistened with spring
water) about the size of the focal salamander.

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On day 1 of the familiar intruder trials, we placed

each focal male (n = 26) in a clear Nunc bioassay dish
(24 24 2 cm hereafter called chamber) containing
a paper towel moistened with spring water and covered
with a transparent lid. We allowed 5 days for the salamander to establish a territory (Jaeger et al., 1986; Wise
and Jaeger, 1998). On day 5, we transferred an intruder
(A) into the focal salamanders chamber for 15 minutes
and allowed the two salamanders to interact. In all four
experiments intruders were transferred into the focal
salamanders chamber via a clear plastic tube to reduce
the stress of handling. After 15 minutes, we removed
intruder (A). On day 10, we introduced into the chamber
the same familiar male intruder (A) that the focal
salamander had seen on day 5, and we recorded the behavior of the focal salamander for 15 minutes.
Focal salamanders in the unfamiliar intruder treatment went through similar procedures as salamanders in
the familiar intruder treatment. However, on day 5, we
placed a different intruder (B) into the focal salamanders chamber for 15 min and then removed intruder (B).
On day 10, instead of reintroducing intruder (B) to the
focal salamander, we introduced a different unfamiliar
intruder (C) for 15 minutes and recorded the behavior of
the focal salamander.
For surrogate control trials, the procedures followed
those of familiar and unfamiliar trials above, except that
on day 5, we placed a different intruder (D) into the
focal salamanders chamber for 15 minutes and then
removed intruder (D). On day 10, we introduced a surrogate to the focal salamander and recorded the behavior of the focal salamander. Intruder salamanders were
only used once in this experiment so that each focal
salamander interacted with a different intruder A, B, C,
and D.
To control for inter-observer bias, the same person
tested the same focal male in all treatments. During each
observation, we recorded the following behavior patterns: time (in seconds) spent in an aggressive posture,
(ATR) the all-trunk raised threat posture (Jaeger and
Schwarz, 1991), and the number of bites administered
by the focal salamander to the intruder (Jaeger, 1984).
Theses bites were all nips that did not break the skin of
any intruder.
1.3 Experiment 2: 8-hour exposure and 5-day separation Interval
We collected adult male salamanders from 2022
April 2006 and conducted this experiment between August and September 2006, from 08: 0016: 00 h. We
randomly assigned 40 male salamanders to each of the

KOHN NR et al.: Memory of conspecifics in male salamanders

following four groups: focal salamanders, future familiar intruders, future unfamiliar intruders, and the partner for the unfamiliar intruder (unfamiliar pair: see
methods below) so that no intruder salamander was used
in more than one treatment. Each focal salamander was
then tested in two treatments: familiar and unfamiliar
intruder trials. After the focal salamander went through
the first treatment, we waited a week before starting the
focal salamanders in the second treatment. We randomly
selected half of the focal salamanders to receive familiar
intruder treatments first and the other half to receive
unfamiliar treatments first. We eliminated the surrogate
treatment from experiments 24, because after experiment 1 we were aware that salamanders would respond
more aggressively towards live intruders than inanimate
objects. Therefore, it was not necessary to continue this
treatment.
On day 1, for familiar intruder trials, we placed the
focal salamander into a chamber containing a moistened
(with spring water) paper towel. On day 5, we introduced the familiar intruder into the focal salamanders
chamber for 8 h. After 8 h, we removed the now familiar intruder and changed the focal salamanders substrate. On day 9, we changed the focal salamanders
substrate again. On day 10, we reintroduced the familiar
male intruder into the focal salamanders chamber, and
recorded the behavior of the focal salamander for 15
min.
Focal salamanders in the unfamiliar intruder trials
went through the same procedures as in familiar intruder
trials (above), except on day 10, when we introduced an
unfamiliar intruder into the focal salamanders chamber.
Unfamiliar intruders were given the same social experience as familiar intruders, as each unfamiliar intruder interacted with a male it had never seen before
(unfamiliar pair) for 8 hours on day 5 (the same time
that focal salamanders and familiar intruders were together). We then introduced the unfamiliar intruder to
the focal salamander on day 10 and recorded the behavior of the focal salamander. All substrate changes were
the same as in familiar intruder trials. We recorded the
same behavioral patterns (ATR and number of bites) as
in experiment 1.
1.4 Experiment 3: 5-day exposure and 5-day separation Interval
We conducted experiment 3 between April and May
2005, from 10: 0016: 00 h, using the same salamanders
as in experiment 1, but those males that were focals in
experiment 1 were not used as focals in experiment 3.
Also, salamanders that interacted with each other in

329

experiment 1 were not paired together in experiment 3.

We randomly assigned 40 male salamanders to each of
the following four groups: focal salamanders, future
familiar intruders, future unfamiliar intruders, or the
partner for the unfamiliar intruder, unfamiliar pair.
Each focal salamander was then tested in two treatments:
1) familiar and 2) unfamiliar intruders. After the focal
salamander went through the first treatment, we waited
a week before starting the focal salamanders in the second treatment. We randomly selected half of the focal
salamanders to receive familiar intruder treatments first
and half to receive unfamiliar intruder treatments first.
In the middle of the experiment, one focal salamander
died from natural causes; therefore we removed its data
from the analysis.
On day 1 of the familiar intruder trials, we placed
each focal salamander and its designated familiar intruder into a chamber containing a paper towel moistened with spring water and covered with a transparent
lid. Both salamanders were allowed 5 days to become
familiar with each other. On day 5, we removed the familiar intruder from the focal salamanders chamber and
placed the familiar intruder into a separate, clean chamber. We also changed the substrate in the focal salamanders chamber. On day 9, we changed the focal
salamanders substrate. On day 10, we introduced the
familiar intruder into the chamber of the focal salamander and recorded the behavior of the focal salamander
for 15 minutes.
In the unfamiliar intruder trials, focal salamanders
went through the same procedures as in familiar intruder
trials (above), except on day 10, when an unfamiliar
intruder was introduced into the focal salamanders
chamber. In order for unfamiliar intruders to have the
same social experience as familiar intruders, each unfamiliar intruder interacted for 5 days with a male it had
never seen before, unfamiliar pair, (the same time that
focal salamanders and familiar intruders were together),
and were separated on day 5. We then introduced the
unfamiliar intruder to the focal salamander on day 10.
We changed the substrates as we did in the familiar intruder trials and recorded the same behavior patterns as
in the previous experiments.
1.5 Experiment 4: 5-day exposure and 15-day separation interval
We collected adult male salamanders from 1315
October 2005. All collection and maintenance procedures were the same as in experiment 3. We conducted
experiment 4 from October to December 2005 from 09:
0016: 00 h. We randomly assigned 40 males to each of

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the following four groups: focal salamanders, future

familiar intruders, future unfamiliar intruders, and the
partner for the unfamiliar intruder (unfamiliar pair: see
rationale in experiments 2 and 3). Again, intruders were
only used once. We tested each focal salamander in two
treatments: 1) familiar and 2) unfamiliar intruder trials.
After the focal salamander went through the first treatment, we waited a week before starting the focal salamanders in the second treatment. We randomly selected
half the focal salamanders to receive the familiar intruder treatment first and half to receive the unfamiliar
intruder treatment first.
In familiar intruder trials, procedures for days 1
through 5 were identical to those for experiment 3. On
day 19, we changed the substrate in the focal salamanders chamber. On day 20, we reintroduced the familiar
intruder into the focal salamanders chamber and recorded the focal salamanders behavior for 15 minutes.
Focal salamanders in the unfamiliar intruder trials
went through the same procedures as in familiar intruder
trials (above), except on day 20 when we introduced an
unfamiliar intruder into the focal salamanders chamber.
All substrate changes were the same as in familiar intruder trials. We recorded the same behavior patterns as
in the first three experiments.
1.6 Statistical analyses
For experiment 1 (15-min exposure and 5-day separation interval), we used the nonparametric Friedman
two-way analysis of variance by ranks test (Siegel and
Castellan, 1988) because we used a within-subjects design and our data were not normally distributed. We
compared the behavior of focal male salamanders across
the three treatments ( = 0.05). If the Friedman test
yielded a significant difference, we then performed
post-hoc tests to compare the three treatments. The
post-hoc comparison automatically adjusts alpha (Siegel
and Castellan, 1988: p. 182). Because we used a withinsubjects design and were examining memory, we also
tested for a treatment-order effect to determine if a
salamanders experience from earlier trials had an effect
on the focal salamanders behavior in subsequent trials.
For experiments 2 (8-hour exposure and 5-day separation interval) and 3 (5-day exposure and 5-day separation interval), we conducted two-tailed Wilcoxon signed
ranks tests (Siegel and Castellan, 1988) to compare time
in ATR, of focal salamanders between the two treatments, with = 0.05. We also tested for treatment order
effects to determine if the salamanders experience from
the first trial had an effect on the second trial. We used
the two-tailed Wilcoxon sign ranks test because we had

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no a priori prediction whether aggression would increase or decrease between treatments. Since too few
bites occurred for statistical analysis, in experiments
13, they are not mentioned further.
For experiment 4 (5-day exposure and 15-day separation interval), we used a two-tailed Wilcoxon signed
ranks test to compare both time in ATR and number of
bites by focal salamanders between the two treatments.
We adjusted to 0.025 because time in ATR and number of bites are both aggressive behaviors. We also
tested for a treatment-order effect.

Results

For experiment 1 (15-min exposure and 5-day separation interval), focal males differed significantly in
ATR behavior across the three treatments (Fr = 16, n =
26, df = 2, P < 0.001) (Fig. 1). Using post hoc comparisons we found that focal salamanders spent significantly
more time threatening both familiar intruders and unfamiliar intruders than threatening the surrogate controls.
No significant differences occurred in ATR threat posture towards familiar and unfamiliar intruders. Also, we
found no significant treatment order effects for ATR in
any of the four experiments.
For experiments 2 (8-hour exposure and 5-day separation interval) and 3 (5-day exposure and 5-day separation interval), focal male salamanders spent significantly more time threatening unfamiliar male intruders
than familiar male intruders (Experiment 2: Wilcoxon

Fig. 1

Time in ATR for Experiment 1

Box and Whisker plots of the behavioral responses of focal male

salamanders. Time in ATR threat posture toward familiar male intruders, unfamiliar male intruders, and surrogates after a 15-min exposure
and 5-day separation interval. The Freidman analysis of variance by
ranks test yielded an overall significant difference; lines above the
treatments represent significant post-hoc comparisons. Boxes represent the inter-quartile ranges, horizontal bars in the boxes represent the
medians, whiskers represent the farthest data points that are not outliers, and the black stars above the whiskers represent the outliers.

KOHN NR et al.: Memory of conspecifics in male salamanders

Fig. 2

Time in ATR for Experiment 2

Box and Whisker plots of the behavioral responses of focal male

salamanders. Time in ATR threat posture toward familiar male intruders and unfamiliar male intruders after an 8-hour exposure and 5-day
separation interval. The line above the treatments represents a significant difference.

Fig. 3

Time in ATR for Experiment 3

Box and Whisker plots of the behavioral responses of focal male

salamanders. Time in ATR threat posture toward familiar and unfamiliar male intruders after a 5-day exposure and 5-day separation interval. The line above the treatments represents a significant difference.

T+ = 529.5, z = 3.09, n = 40, n = 36 non-tied scores, P =

0.002, Fig. 2; Experiment 3: Wilcoxon T+ = 569, z =
4.16, n = 39, n = 35 non-tied scores, P < 0.00006, Fig. 3).
For experiment 4 (5-day exposure and 15-day separation interval), we found no significant difference in
ATR threat posture toward unfamiliar male intruders
compared to familiar male intruders (Wilcoxon T+ =
448.5, z = 1.46, n = 40, n = 37 non-tied scores, P =
0.144, Fig. 4). Also, we found no significant difference
in number of bites towards familiar male intruders
(mean = 1.43, median = 0, range 018) and unfamiliar
male intruders (mean = 0.68, median = 0, range 09)
(Wilcoxon T+ = 57.5, z = 1.64, n = 40, n = 12 non-tied
scores, P = 0.101, adjusted = 0.025).

3 Discussion
Red-backed salamanders at our research site defend

Fig. 4

331

Time in ATR for Experiment 4

Box and Whisker plots of the behavioral responses of focal male

salamanders. Time in ATR threat posture toward familiar and unfamiliar male intruders after a 5-day exposure and 15-day separation
interval.

multi-purpose territories. Through four experiments, we

tested three different exposure durations (15 minutes, 8
hours, and 5 days) and two different separation intervals
(5 days and 15 days). In experiments 2 and 3, when the
exposure durations were 8 hours and 5 days, respectively, and the separation interval was 5 days, males
were more aggressive toward unfamiliar intruders than
familiar intruders. In contrast, in experiments 1 (15-min
exposure, 5-day separation) and 4 (5-day exposure,
15-day separation), male salamanders did not act more
aggressively toward unfamiliar intruders than familiar
intruders. These results indicate that male red-backed
salamanders can remember familiar conspecifics (e.g.,
territorial neighbors) when the exposure duration is
relatively long (e.g., 8 hours and longer) and the separation interval is relatively short (e.g., 5 days). This recognition memory may be particularly important for
red-backed salamanders at our research site at MLBS,
where the population density on the forest floor is about
2.8 salamanders/m2 and 49% of the population consists
of transient individuals that do not hold territories
(Mathis, 1991). Thus, territorial intruders are probably
frequent, and aggressive (Wise and Jaeger, 1998).
However a lack of differences in behavior toward
familiar and unfamiliar intruders does not directly indicate a lack of recognition memory. Memory is a neural
representation of a past experience that can only be assessed by a change in behavior indicating that the animal has remembered that experience (Kawecki, 2010).
Therefore, in our results, a lack of differences in behavior could occur either because the focal salamanders
did not remember the familiar conspecifics, or because
they did remember the familiar conspecifics but it was
no longer beneficial to behave differently towards them.

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The distinction between recognition memory and

changes in behavior are particularly important in experiment 4. Here, after 15 days of separation, the focal
salamanders either did not remember the previously
familiar conspecifics or did remember them but treated
them as aggressively as if they were unfamiliar strangers. If a salamander leaves its territory for 15 days, and
then returns, other floater salamanders may take over
the territory (Mathis, 1990); thus, the returning salamander poses a threat and new boundaries may need to
be established.
Territorial defense has the potential to be costly, as it
increases the chance of injury during escalated contests
(Jaeger, 1981); increases energy spent advertising and
defending territories that could be allocated to other
activities; and increases time spent fighting that could
be used for finding food, mates, or avoiding predators
(Davies, 1978; Hardouin et al., 2006; Golabek et al.,
2012). Yet not all conspecific intruders pose the same
threat (Davies, 1978). Transient individuals searching
for a territory pose more of a threat than established
territorial neighbors or juveniles (Palphramand and
White, 2007). One way to decrease the cost of territorial
defense is to recognize and remember familiar individuals/neighbors from unfamiliar individuals/strangers
(Hardouin et al., 2006). This recognition can save physiological costs by minimizing the energy expended on
aggressive acts, by preventing escalated contests between familiar individuals, and by decreasing time lost
to fighting (Hardouin et al., 2006).
One variable that might affect how a territorial owner
acts towards a conspecific intruder is the length of time
that the intruder is in or near the owners territory. If
the intruder is in an owners territory for a short amount
of time (e.g., 15-min exposure), simulating a transient or
floater individual moving through a territory, it may not
be beneficial and/or long enough for a territorial owner
to recognize, remember, and therefore decrease its aggression toward that intruder. However, if the individual
remains for a longer time period (e.g., 8-hour and 5-day
exposure), then it may become beneficial to remember,
recognize, and treat that intruder as familiar or a
neighbor, and therefore decrease aggression toward that
individual which decreases the potential cost of being
territorial.
Using a 5-day exposure and 5-day separation, Kohn
and Jaeger (2009) found that red-backed salamanders
can use only chemical cues or only visual cues to remember familiar individuals even without direct intruder interactions. Therefore, in at least our 5-day ex-

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posure and 5-day separation experiment, our results are

based on recognition memory of individuals and not just
a response to the behavior of intruders. What we do not
know is the type of visual cues used by red-backed
salamanders for individual recognition (e.g., morphological characteristics: Van Dyk and Evans, 2007).
Comparing visual cue recognition in red-backed salamander populations near MLBS that do not have such
variation in color morphs would help us determine
whether or not recognition is based more on behavioral
cues or color patterns.
Previous studies suggest that chemical cue recognition in red-backed salamanders is stronger than visual
cue recognition (Page and Jaeger, 2004; Kohn and Jaeger, 2009). This is not surprising considering that other
salamanders and tadpoles predominantly use chemical
cues over other cues for recognizing conspecifics, kin,
and predators (e.g., Blaustein and Waldman, 1992;
Wildy and Blaustein, 2001; Waldman and Bishop, 2004;
Thaker et al., 2006; Epp and Gabor, 2008). In order to
better understand individual recognition versus other
forms of recognition (Tibbetts and Dale, 2007) in
red-backed salamanders, conducting chemical profiles
on individual skin secretions (pheromones) to determine
if individual salamanders differ in their chemical profiles would be useful. Red-backed salamanders have
several secretory sources (Simons and Felgenhauer,
1992), including fecal pellets that vary in individual
chemical profiles based on diet (Walls et al., 1989). Secretions from the postcloacal gland are important in
territorial defense, and these secretions may possibly
provide individually unique signals (Simons and Felgenhauer, 1992; Jaeger and Gabor, 1993). Chemical
profiles have been made on two ant species, Pachycondyla villosa and Lasius niger, where the chemical
profiles of queens varied in hydrocarbon relative abundance from individual to individual, yet individual recognition only occurred in Pachycondyla villosa which
forms dominance hierarchies (DEttorre and Heinze,
2005; Drier and DEttorre, 2009). Alternatively, salamanders could mark familiar individuals with their own
unique chemical signatures which would allow recognition of familiar individuals in subsequent encounters
without the need to remember specific individuals (Ivy
et al., 2005).
In this population of red-backed salamanders, we
suggest that natural selection has acted on individuals to
recognize long-term territorial intruders or neighbors
from short-term transient individuals. However, not all
populations of red-backed salamanders are territorial

KOHN NR et al.: Memory of conspecifics in male salamanders

(Quinn and Graves, 1999). To better understand individual recognition and territoriality in this species one
needs to examine the environmental conditions where
territoriality exists so that one can then examine the
social interactions, phenotypic variation, and recognition signals needed for individual recognition to occur.
Acknowledgements We thank T. Williams and S. Nguyen
for help running experiments; H. Wilbur and E. Nagy for permission to conduct research at Mountain Lake Biological Station; B. Dantzer, D. McCauley, J. Fuller and her parents for
help in collecting salamanders; T. Kohn for her help in maintaining the salamanders; A. Aspbury, J. Burns, C. Gabor, M.
Kalish, P. Klerks, P. Leberg, and B. Moon for comments on
previous versions of the manuscript. This research was partly
funded by the Graduate Student Organization and the Biology
Department of The University of Louisiana at Lafayette, and
by a Louisiana Board of Regents Doctoral Fellowship
(LEQSF (2003-08)-GF-31 to N.R.K. through R.G.J.). J.M.D.
was supported by a USDA IFAFS Multidisciplinary Graduate
Education Training Grant (2001-52100-11506). Salamanders
were collected under permit 020999 during October 2004 and
permit 026118 during October 2005 and April 2006 from the
Virginia Department of Game and Inland Fisheries. Laboratory protocols were approved by The University of Louisiana
at Lafayette Animal Care and Use Committee (IACUC No.
2003-8717-094, No. 2004-8717-077, and No. 2005-8717123).

References
Alberici da Barbiano L, Rangel L, Aspbury AS, Gabor CR, 2012.
Male permissiveness in a unisexual-bisexual mating complex
promotes maintenance of a vertebrate unisexual spermdependent species. Behaviour 149: 869879.
Alberts AC, Werner DI, 1993. Chemical recognition of unfamiliar
conspecifics by green iguanas: Functional significance of different signal components. Anim. Behav. 46: 197199.
Aragon P, Lopez P, Martin J, 2001. Chemosensory discrimination
of familiar and unfamiliar conspecifics by lizards: Implications
of field spatial relationships between males. Behav. Ecol. Sociobiol. 50: 128133.
Bee MA, Gerhardt HC, 2002. Individual voice recognition in a
territorial frog Rana catesbeiana. P. R. Soc. B. 269: 1443
1448.
Bee MA, Kozich CE, Blackwell KJ, Gerhardt HC, 2001. Individual variation in advertisement calls of territorial male green
frogs Rana clamitans: Implications for individual discrimination. Ethology 107: 6584.
Blaustein AR, Waldman B, 1992. Kin recognition in anuran amphibians. Anim. Behav. 44: 207221.
Botero CA, Riveros JM, Vehrencamp SL, 2007. Relative threat
and recognition ability in the responses of tropical mockingbirds to song playback. Anim. Behav. 73: 661669.
Carazo P, Font E, Desfilis E, 2008. Beyond 'nasty neighbours' and
'dear enemies'? Individual recognition by scent marks in a lizard Podarcis hispanica. Anim. Behav. 76: 19531963.

333

Cowan N, 2008. What are the differences between long-term,

short-term, and working memory? In: Sossin WS, Lacaille J-C,
Castellucci VF, Belleville S ed. Progress in Brain Research.
The Netherlands: Elsevier B.V., 323338.
Davies NB, 1978. Ecological questions about territorial behaviour.
In: Krebs JA, Davies NB ed. Behavioral Ecology: An Evolutionary Approach. 1st edn. Sinauer Associates: Massachusetts,
USA, 317350.
DEttorre P, Heinze J, 2005. Individual recognition in ant queens.
Curr. Biol. 15: 21702174.
Dreier S, DEttorre P, 2009. Social context predicts recognition
systems in ant queens. J. Evol. Biol. 22: 644649.
Dukas R, 1998. Constraints on information processing and their
effects on behavior. In: Dukas R ed. Cognitive Ecology: The
Evolutionary Ecology of Information Processing and Decision
Making. Chicago: University of Chicago Press, 89127.
Dukas R, 2004. Evolutionary biology of animal cognition. Annu.
Rev. Ecol. Evol. Syst. 35: 347374.
Epp KJ, Gabor CR, 2008. Innate and learned predator avoidance
mediated by chemical cues in Eurycea nana. Ethology.
114:607615.
Freas CA, LaDage LD, Roth III TC, Pravosudov VV, 2012. Elevation-related differences in memory and the hippocampus in
mountain chickadees Poecile gambeli. Anim. Behav. 84:
121127.
Gabor CR, 1995. Correlational test of Mathis hypothesis that
bigger salamanders have better territories. Copeia. 1995:
729735.
Gherardi F, Cenni F, Parisi G, Aquiloni L, 2010. Visual recognition of conspecifics in the American lobster Homarus americanus. Anim. Behav. 80: 713719.
Gillette JR, Peterson MG, 2001. The benefits of transparency:
Candling as a simple method for determining sex in redbacked salamanders Plethodon cinereus. Herpetol. Rev. 32:
233235.
Gillette JR, Jaeger RG, Peterson MG, 2000. Social monogamy in a
territorial salamander. Anim. Behav. 59: 12411250.
Glinski TH, Krekorian CO, 1985. Individual recognition in
free-living adult male desert iguanas Dipsosaurus dorsalis. J.
Herpetol. 19: 541544.
Golabek KA, Ridley AR, Radford AN, 2012. Food availability
affects strength of seasonal territorial behaviour in a cooperatively breeding bird. Anim. Behav. 83: 613619.
Hardouin LA, Tabel P, Bretagnolle V, 2006. Neighbour-stranger
discrimination in the little owl Athena noctua. Anim. Behav.
72: 105112.
Horne EA, Jaeger RG, 1988. Territorial pheromones of female
red-backed salamanders. Ethology 78: 143152.
Ivy TM, Weddle CB, Sakaluk SK, 2005. Females use self-referent
cues to avoid mating with previous mates. P. R. Soc. B. 272:
24752478.
Jaeger RG, 1980a. Fluctuation in prey availability and food limitation for a terrestrial salamander. Oecologia. 44: 335341.
Jaeger RG, 1980b. Microhabitats of a terrestrial forest salamander.
Copeia 1980: 265268.
Jaeger RG, 1981. Dear enemy recognition and the costs of aggression between salamanders. Am. Nat. 117: 962974.
Jaeger RG, 1984. Agonistic behavior of the red-backed salamander.
Copeia. 1984: 309314.

334

Current Zoology

Jaeger RG, Forester DC, 1993. Social behavior of plethodontid

salamanders. Herpetologica 49: 163175.
Jaeger RG, Gabor CR, 1993. Intraspecific chemical communication by a territorial salamander via the postcloacal gland.
Copeia. 1993: 11711174.
Jaeger RG, Gillette JR, Cooper RC, 2002. Sexual coercion in a
territorial salamander: Males punish socially polyandrous female partners. Anim. Behav. 63: 871877.
Jaeger RG, Goy JM, Tarver M, Marquez CE, 1986. Salamander
territoriality: Pheromonal markers as advertisement by males.
Anim. Behav. 34: 860864.
Jaeger RG, Schwarz JK, 1991. Gradational threat postures by the
red-backed salamander. J. Herpetol. 25: 112114.
Johnston RE, Bullock TA, 2001. Individual recognition by use of
odours in golden hamsters: the nature of individual representations. Anim. Behav. 61: 545557.
Joseph LL, Cope CT, Kohn NR, Jaeger RG, 2005. The influence
of familiarity on socially monogamous behavior of red-backed
salamanders. Herpetologica. 61: 343348.
Kawecki TJ, 2010. Evolutionary ecology of learning: Insights
from fruit flies. Popul. Ecol. 52: 1525.
Kohn NR, Jaeger RG, 2009. Male salamanders remember individuals based on chemical or visual cues. Behaviour 146:
14851498.
Lang C, Jaeger RG, 2000. Defense of territories by male-female
pairs in the red-backed salamander Plethodon cinereus. Copeia.
2000: 169177.
Mateo JM, 2006. Development of individually distinct recognition
cues. Dev. Psychobiol. 48: 508519.
Mathis A, 1989. Do seasonal spatial distributions in a terrestrial
salamander reflect reproductive behavior or territoriality?
Copeia 1989: 788791.
Mathis A, 1990. Territoriality in a terrestrial salamander: The
influence of resource quality and body size. Behaviour 112:
162174.
Mathis A, 1991. Territories of male and female terrestrial salamanders: Costs, benefits, and intersexual spatial associations.
Oecologia. 86: 433440.
McGavin M, 1978. Recognition of conspecific odors by the salamander Plethodon cinereus. Copeia 1978: 356358.
McLeman MA, Mendl M, Jones RB, White R, Wathes CM, 2005.
Discrimination of conspecifics by juvenile domestic pigs Sus
scrofa. Anim. Behav. 70: 451461.
Menzel R, Manz R, Greggers U, 2001. Massed and spaced learning in honeybees: The role of CS, US, the intertrial interval,
and the test interval. Learn. Memory 8: 198208.
Myrberg Jr. AA, Riggio RJ, 1985. Acoustically mediated individual recognition by a coral reef fish Pomacentrus partitus.
Anim. Behav. 33: 411416.
Nairne JS, 2002. Remembering over the short-term: The case
against the standard model. Annu. Rev. Psychol. 53: 5381.
Odling-Smee LC, Boughman JW, Brathwaite VA, 2008. Sympatric species of threespine stickleback differ in their performance in a spatial learning task. Behav. Ecol. Sociobiol. 62:
19351945.

Vol. 59

No. 3

Page RB, Jaeger RG, 2004. Multimodal signals, imperfect information, and identification of sex in red-backed salamanders
Plethodon cinereus. Behav. Ecol. Sociobiol. 56: 123139.
Palphramand KL, White PCL, 2007. Badgers Meles meles discriminate between neighbors, alien, and self scent. Anim. Behav. 74: 429436.
Quinn VS, Graves BM, 1999. Space use in response to conspecifics by the red-backed salamander (Plethodon cinereus,
Plethodontidae, Caudata). Ethology 105: 9931002.
Ruiz JC, Soler MJ, Dasi C, 2004. Study time effects in recognition
memory. Percept. Motor. Skill. 98: 638642.
Ryan MJ, Rand AS, 1993. Species recognition and sexual selection as a unitary problem in animal communication. Evolution
47: 647657.
Sayler A, 1966. The reproductive ecology of the red-backed salamander Plethodon cinereus in Maryland. Copeia 1966: 183193.
Sheehan MJ, Tibbetts EA, 2008. Robust long-term social memories in a paper wasp. Curr. Biol. 18: R851R852.
Sheehan MJ, Tibbetts EA, 2010. Selection for individual recognition and the evolution of polymorphic identity signals in
Polistes paper wasps. J. Evol. Biol. 23: 570577.
Shettleworth SJ, 1998. Cognition, Evolution and Behavior. New
York: Oxford University Press.
Shettleworth SJ, 2001. Animal cognition and animal behaviour.
Anim. Behav. 61: 277286
Siegel S, Castellan Jr. NJ, 1988. Nonparametric Statistics for the
Behavioral Sciences. 2nd edn. New York: McGraw-Hill.
Simons RR, Felgenhauer BE, 1992. Identifying areas of chemical
signal production in the red-backed salamander Plethodon
cinereus. Copeia 1992: 776781.
Temeles EJ, 1994. The role of neighbours in territorial systems:
When are they dear enemies? Anim. Behav. 47: 339350.
Thaker M, Gabor CR, Fries JN, 2006. Sensory cues for conspecific associations in aquatic San Marcos salamanders. Herpetologica. 62: 151155.
Tibbetts EA, 2002. Visual signals of individual identity in the
wasp Polistes fuscatus. P. R. Soc. B. 269: 14231428.
Tibbetts EA, Dale J, 2007. Individual recognition: It is good to be
different. Trends. Ecol. Evol. 22: 529537.
Van Dyk DA, Evans CS, 2007. Familiar-unfamiliar discrimination
based on visual cues in the Jacky dragon Amphibolurus muricatus. Anim. Behav. 74: 3344.
Waldman B, Bishop PJ, 2004. Chemical communication in an
archaic anuran amphibian. Behav. Ecol. 15: 8893.
Waldman B, Frumhoff PC, Sherman PW, 1988. Problems of kin
recognition. Trends. Ecol. Evol. 3: 813.
Walls SC, Mathis A, Jaeger RG, Gergits WF, 1989. Male salamanders with high-quality diets have faeces attractive to females. Anim. Behav. 38: 546548.
Wildy EA, Blaustein AR, 2001. Learned recognition of interspecific predators in larval long-toed salamanders Ambystoma maccrodactylum. Ethology. 107: 479493.
Wise SE, Jaeger RG, 1998. The influence of tail autotomy on
agonistic behaviour in a territorial salamander. Anim. Behav.
55: 17071716.