379

© 2006 Universities Federation for Animal Welfare Animal Welfare 2006, 15: 379-382
The Old School, Brewhouse Hill, Wheathampstead, ISSN 0962-7286
Hertfordshire AL4 8AN, UK

Long-term social memory in the laboratory rat (Rattus norvegicus)

OHP Burman* and M Mendl

Division of Farm Animal Science, Department of Clinical Veterinary Science, University of Bristol, Langford, Bristol BS40 5DU, UK
* Contact for correspondence and request for reprints: oliver.burman@bristol.ac.uk

Abstract
A key question in the management of group-housed captive animals is how long can an individual be removed from a social group
and still be reintroduced with minimal social upheaval. In order to answer this question we require a knowledge of how long cage-
mates, following a specified period of group-housing, can remember one another after separation. This issue was investigated in labo-
ratory rats (Rattus norvegicus). Rats were group-housed for 18 days before being housed individually. One hour, 48 hr, and 96 hr
after separation, they were exposed simultaneously to odour cues originating from unfamiliar rats and from former cage-mates. The
rats spent significantly more time investigating the unfamiliar odour 1 hr and 48 hr, but not 96 hr, after separation, suggesting that,
after 18 days of group-housing, juvenile rats remember former cage-mates for between 48 and 96 hr. The implications of this result
for animal welfare are discussed.

Keywords: animal welfare, habituation/discrimination, laboratory rats, long-term memory, social memory, social recognition

Introduction experimental purposes, and, second, established tests exist

Research into different areas of animal cognition eg studies
of social memory and recognition (Burman & Mendl 2000,
2004), spatial memory (Laughlin & Mendl 2000, 2004),
visual perspective taking (Held et al 2001a,b), not only
furthers our understanding of the fundamental cognitive
processes of non-human animals, but also provides us with
information that may help to improve animal husbandry and
welfare (Nicol 1996; Mendl et al 2001).
For example, because the majority of group housed captive
animals are kept together for extended periods (eg weeks:
broiler chickens; months: laboratory rodents; years: zoo
animals), studies of long-term social memory abilities may
provide important information on how best to manage and
control husbandry and housing procedures that disrupt and
reunite established social groupings (Mendl 1999). An
increase in aggression, and therefore an apparent reduction
in welfare, can occur when animals are reunited after a
period of separation (Ewbank & Meese 1971). Thus, if an
animal is to be removed from a stable social group for
husbandry or experimental reasons, it is important to know
for how long that individual can be separated from its
group-mates while still allowing safe reintroduction. In
order to answer this question we need to know for how long,
following a given period of group-housing, former cage-
mates can remember one another after separation.
We investigated this issue using laboratory rats because,
first, they tend to be housed in groups for several weeks and
can be separated from their social groups temporarily for
for assessing social memory in laboratory rats (Thor &
Holloway 1982; Engelmann et al 1995). Whilst short-term
memory for conspecifics, typically over periods of 30 - 120
min, has been the focus of much research (Thor &
Holloway 1982; Dantzer et al 1987; Hlinak & Krejci 1991;
Gheusi et al 1994; Engelmann et al 1995), comparatively
little attention has been directed towards long-term social
memory. Using spontaneous behaviour tests (eg the habitu-
ation/discrimination technique [see later]), researchers have
shown that rats appear unable to remember conspecifics for
longer than two hours (Bluthé & Dantzer 1990; Taylor et al
1999) when the initial encounter with the to-be-remembered
conspecific is around 5 min long. Even when rats received
six × 5 min encounters each separated by 10 min intervals
they appeared unable to remember conspecifics 24 h later
(Sekiguchi et al 1991).
However, it may not be realistic to expect rats to remember
conspecifics long-term when they have only had the oppor-
tunity to encounter those conspecifics for such a brief initial
period of time. Such studies also do not reflect real-life
husbandry procedures, whereby animals are often group-
housed for several days before any individuals are
temporarily removed. We decided, therefore, to investigate
long-term social memory in laboratory rats by housing indi-
viduals together for 18 days, and then assessing whether or
not, following this extended familiarisation period, the rats
were subsequently able to remember former cage-mates
after separation – from one hour up to four days.

Universities Federation for Animal Welfare Science in the Service of Animal Welfare

https://doi.org/10.1017/S0962728600030712 Published online by Cambridge University Press

 380 Burman and Mendl
The results of this study have direct implications for the
welfare of laboratory rats, and, if we consider the rat as a
model species, may also have implications for the welfare
of other group-housed captive species.
Materials and methods
Subjects, housing, and care
We used 16 unrelated (no shared parent) female (21 days
old at the start of experiment) Lister hooded rats (Harlan
UK Limited, Bicester, UK). Unrelated rats were used to
ensure that it was familiarity per se that was examined; with
no confounding influence of kin recognition. We used
juvenile rats in order to avoid any potentially confounding
effects of dominance on performance in the
habituation/discrimination technique (Carr et al 1976;
Brown 1992). Previous research has suggested no differ-
ence between the sexes in pre-maturation social memory
performance (Thor & Holloway 1982). All the rats were
housed in the same room in which the experiments took
place, with temperature 19 ± 1°C, relative humidity 46%
and a reverse dark-light cycle (light on from 1900 - 0700h).
Observations were made in the dark phase of the cycle, with
dim ‘white’ light (10 watt) provided during experimenta-
tion. Home cages, (50 × 33 × 21 cm)
(length × width × height), contained sawdust bedding, an
enrichment toy and ad libitum food (Harlan Teklad
Laboratory Diet) and water. We identified individual rats
from natural variation in coat colour markings. Disposable
gloves were worn when handling rats to restrict odour
transfer. This experiment was carried out with a University
of Bristol investigation number (UB/98/L038), as deter-
mined by the University of Bristol Home Office Liaison
Team (HOLT).
The habituation/discrimination technique
We used the habituation/discrimination technique
(Johnston & Jernigan 1994; Johnston & Bullock 2001).
This procedure involves repeated presentation of a
stimulus (eg odour cue) obtained from a specific indi-
vidual to a subject animal. The behavioural response of
the subject towards this stimulus, usually investigation, is
recorded, and generally decreases with each presentation.
Following this habituation, which is taken to indicate the
subject’s familiarity with the stimulus, odour stimuli from
both the original individual and a novel individual are
then introduced simultaneously. Successful discrimina-
tion between the two stimuli, in terms of significantly
more time spent investigating the novel odour stimulus,
indicates continued habituation for, and thus recogni-
tion/memory of, the original stimulus.
This technique should overcome any potentially
confounding motivational changes between the introduction
and subsequent reintroduction of the same stimuli (Johnston
1993) that could, for example, result in a general elevation
or decrease in response levels that mask/accentuate habitu-
ation or dishabituation. Therefore, regardless of motivation,
it is the comparison between the level of investigation of the
© 2006 Universities Federation for Animal Welfare
https://doi.org/10.1017/S0962728600030712 Published online by Cambridge University Press
two stimuli within each test, rather than between tests, that
is important. In this study, rather than repeatedly presenting
the odour of one individual to another to induce habituation
and the presumed familiarity of that odour; familiarity, and
hence habituation, were assumed to have occurred
following the group housing of subject and stimulus
animals for an extended period of time (Epple & Niblick
1997; Mateo & Johnston 2000).
A potential problem with using an habituation/discrimina-
tion technique of this kind is that there can be difficulty with
the interpretation of ‘negative’ results. Whilst a statistically
significant preference to investigate the novel stimulus, ie
discrimination, can be interpreted relatively reliably as a
recognition of, and habituation to, the original stimulus, any
failure to show discrimination should be interpreted with
caution. For example, a negative result should be described
as ‘a failure to demonstrate discrimination’ rather than ‘a
failure to discriminate’ per se.
Testing procedure
At 21 days of age the rats were housed in four groups of
four for 18 days, to allow them to become familiar with the
other members of their particular group. They were then
separated and housed individually for the duration of testing
(a total period of four days) after which they were group-
housed for use in a further experiment. Half the rats were
randomly selected as subjects (n = 8) and the remainder as
odour donors. The subjects were tested at one hour, 48 h,
and 96 h post separation. Tests were carried out in the home
cage, and involved the simultaneous presentation of unfa-
miliar and familiar (former cage-mate) odour cues for
10 min. Significantly more time spent investigating the
unfamiliar odour was taken to indicate successful recogni-
tion of the familiar odour (Johnston 1993). For each of the
three tests, odour cues from different familiar and unfa-
miliar rats were used, so that subjects never encountered the
same odour cues during repeated testing.
Time (seconds) spent investigating the odours was recorded
(ie sniffing, licking, or the subject’s nose being within one
cm of the odour container) using a Psion event recorder
(Psion PLC, London, UK) with ‘Observer’ software
(Noldus Information Technology, Wageningen, Holland).
Odour cues
Previous studies have indicated the suitability of using
odour cues as social stimuli in recognition tests (Carr et al
1976; Sawyer et al 1984; Burman & Mendl 2002). We
obtained the odour samples by placing a sheet of absorbent
paper on the floor of the rats’ individual home-cages for one
hour prior to testing. This ensured that urine samples were
relatively uniform in age. Where the rat had urinated on the
paper (all rats urinated at least once within the hour),
scissors were used to cut out the marked area and this
sample was then placed inside a spherical wire mesh
container (20 cm³). We attached the containers to a test cage
lid that could then be introduced into the home cage of the
subject rat when testing was to start. The two odour samples
were presented at opposite sides of the subjects’ home cage,
 Long-term social memory in rats 381

and these positions (either left or right side) were alternated Figure 1
between different subjects to avoid any effect of side pref-
erence. The odour containers, the test cage lid, and the
scissors were disinfected between each odour collection and
each test, and latex gloves were also used to minimise the
risk of odour transfer between samples.
Data analysis
The total amount of time (seconds) spent investigating each
of the two containers holding the odour samples was
analysed using Minitab version 12 (Minitab Inc,
Philadelphia, USA). The data met requirements for
normality and homogeneity of variance, and so a parametric
repeated measures General Linear Model (GLM) was used
to analyse the data with paired t-tests for post hoc analysis.
Descriptive statistics of normally distributed data are
means ± standard errors.
Results
Differences in the amount of investigation directed towards the
An initial analysis was carried out to compare the levels of
familiar (white columns) and unfamiliar (black columns) odour
total investigation (unfamiliar + familiar odour) between the stimuli by the subjects, at one hour, 48 hours, and four days after
three tests (1 hr, 48 hr, 96 hr). This revealed a significant separation.
difference between the tests (F2,14 = 15.04, P < 0.001), with
post hoc analysis indicating a significant reduction in total
investigation between the 1 hr and 48 hr tests (P < 0.01), but Our study thus demonstrates the spontaneous capability of
no difference between the 48 hr and 96 hr tests. This result rats for long-term social memory following an extended
suggests a general habituation to the test procedure (18 day) period of familiarisation. This suggests that
occurring between the 1 hr and 48 hr tests for both stimuli previous studies may have failed to demonstrate long-term
(familiar: F2,14 = 3.76, P < 0.05; unfamiliar: F2,14 = 14.5, social memory in rats because the initial familiarisation
P < 0.001), but not between the 48 hr and 96 hr tests. period selected was too brief (eg 5 min: Taylor et al 1999).
However, the habituation/discrimination technique used in The results also suggest that, following a familiarisation
this study was selected precisely because it is able to take period of 18 days, juvenile rats removed for at least
such changes in general motivational level into account (see 48 hours might be safely reintroduced into their original
‘Materials and methods’). social group, because it is still possible for them to success-
fully discriminate between the odours of previously familiar
We then carried out a repeated measures GLM with test (1 hr, (former cage-mates) and unfamiliar individuals. It should
48 hr, 96 hr) and odour (unfamiliar/familiar) as factors. This be noted, though, that the ability to successfully discrimi-
revealed a significant interaction between test and odour nate between individuals (or their cues) might not, in itself,
(F2,14 = 5.55, P < 0.01). Post hoc analysis of this test × odour be sufficient to avoid further aggression that may result as a
interaction revealed that both one hour (t8 = 4.27, P < 0.01) consequence of additional factors, ie the dominance status
and 48 hr (t8 = 2.59, P < 0.05) after separation, the rats did of the reintroduced individual (Ewbank & Meese 1971).
indeed spend significantly more time investigating the odour
It is also possible that the length of time between encounters
of a novel juvenile than that of a former cage-mate (see
in studies of long-term social memory allows greater oppor-
Figure 1). Thus, although there was evidence of a general
tunity for the disruption of social memory by challenging
habituation to the test procedure between 1 and 48 h, discrim-
environmental events, for example exposure to the odours
ination was apparent at both these time points. However,
of novel individuals during cage cleaning. Nevertheless,
96 hours after separation, by which time there had been no
even though short-term social memory does appear suscep-
further habituation to the test procedure – total levels of
tible to disruption (Burman & Mendl 2000), familiarity with
investigation were no different to those at 48 h – there was
a conspecific gained over an extended period of time (ie
now no evidence of any discrimination between the familiar
18 days) may make memory less vulnerable to disruption
and unfamiliar odours (t8 = 1.21, P = 0.27). (Burman & Mendl 2004).
Discussion Animal welfare implications
The results of this experiment suggest that juvenile female In conclusion, we have described how, following 18 days of
rats are able to remember the identity of former cage-mates group-housing, juvenile rats appear to remember cage-
that they had previously spent 18 days housed with for at mates with whom they were previously housed for at least
least 48 hours following separation but failed to demon- 48 hours, but that similarly successful recognition is no
strate successful recognition when this period of separation longer demonstrated after 96 hours of separation. This
was increased to 96 hours. indicates that juvenile rats that are temporarily separated

Animal Welfare 2006, 15: 379-382

https://doi.org/10.1017/S0962728600030712 Published online by Cambridge University Press

 382 Burman and Mendl

from their cage-mates for experimental or other reasons,
should be reunited them within 48 hours, and before
96 hours. This should maximise the chances of social recog-
nition and a harmonious reintroduction. The results of this
study are therefore an essential first step in developing an
understanding of the cognitive processes that inform the
practical issue of how and when to remove and reintroduce
animals from social groups.
Acknowledgments
This work was funded by a University of Bristol
Postgraduate Scholarship to Oliver HP Burman. The
authors wish to thank two anonymous reviewers for their
helpful suggestions.
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© 2006 Universities Federation for Animal Welfare

https://doi.org/10.1017/S0962728600030712 Published online by Cambridge University Press