Training Nonhuman Primates Using Positive Reinforcement Techniques

Physical cage enrichment—exercise devices for rodents in the laboratory—often includes running wheels. This study compared responses of mice in enriched physical and social conditions and in standard social conditions to wheel running, individual housing, and open-field test. The study divided into 6 groups, 48 female BALB/c mice group housed in enriched and standard conditions. On alternate days, the study exposed 2 groups to individual running wheel cages. It intermittently separated from their cage mates and housed individually 2 groups with no running wheels; 2 control groups remained in enriched or standard condition cages. There were no significant differences between enriched and standard group housed mice in alternate days' wheel running. Over time, enriched, group housed mice ran less. Both groups responded similarly to individual housing. In open-field test, mice exposed to individual housing without running wheel moved more and faster than wheel running and home cage control mice. They have lower body weights than group housed and wheel running mice. Intermittent withdrawal of individual housing affects the animals more than other commodities. Wheel running normalizes some effects of intermittent separation from the enriched, social home cage.     

Behaviour of laboratory mice in different housing conditions when allowed to self-administer an anxiolytic

Standard cages prevent mice from performing several natural behaviours for which they are motivated. As a consequence, abnormal behaviours sometimes develop and mice often spend long periods inactive. To improve welfare, cages are sometimes furnished with items such as nesting material, shelters and running wheels. We have previously reported that when allowed to self-administer an anxiolytic, mice in furnished cages consume less anxiolytic than mice in standard cages. This paper presents the results of behaviour studies of the mice in the same experiment. Female C57BL/6J mice (3 per cage) were housed in Standard (n = 10), Unpredictable (n = 10) or Furnished (n = 6) cages. Unpredictable cages were identical to Standard cages, but were exposed to unpredictable events two to three times a week. Furnished cages were double the size of Standard cages and contained nesting material, nest box, tubes, chew blocks and a running wheel. During three consecutive periods, mice had access to only water (control), water or an anxiolytic solution on a daily alternating schedule (forced consumption), and finally, both water and anxiolytic (self-administration). Behaviour was analysed from video recordings taken during the dark phase. The housing type affected behaviour both under the control and the self-administration conditions. Overall, mice in Furnished cages spent less time resting and performing bar-related behaviours and more time on exploratory/locomotory behaviours. Mice in Furnished cages also performed less bar-circling stereotypies than mice in Standard cages. The Unpredictable treatment did not significantly affect behaviour compared to mice in the Standard conditions. There was an overall effect of anxiolytic availability on rest-related behaviours and on exploration-locomotion behaviours, in that mice rested more and spent less time on exploration and locomotion when they were able to self-administer the anxiolytic.     

The effect of cage size and enrichment on core temperature and febrile response of the golden hamster

The aim of this study is to determine the effect of cage size and cage enrichment. Golden hamsters were individually housed in standard cages of four different sizes and in enriched cages of three different sizes since 3 weeks of age. Each of the seven housing groups consisted of 12 hamsters. After 14 weeks of housing in their respective environments the measurements started. The mean baseline rectal temperature was significantly higher in hamsters housed in small cages than in hamsters housed in large cages. After the injection of fever-inducing lipopolysaccharide rectal temperature increased by 1 to 2 degrees C. The increase of rectal temperature and the fever index were the highest in animals housed in large cages and the smallest in animals housed in small cages. Through cage enrichment and increasing cage size the mean febrile response increased while the mean baseline rectal temperature decreased. Cage size and cage enrichment had no effect on the dispersion of the measured values. The differences in microclimate between large and small cages were too small to have an effect on thermoregulation. The results indicate that housing in small cages induce chronic stress which obviously affects thermoregulation. The findings demonstrate that the results of some physiological experiments are significantly influenced by the pre-experimental housing conditions.     

The effects of running activity on the reproductive axes of rodents

Access to a running wheel causes gonadal recrudescence in Syrian hamsters whose reproductive axes have been suppressed by housing them under short day lengths (Borer et al. 1983). The first experiment tested the generality of this phenomenon in a population of rodents that is genetically heterogeneous for reproductive photoresponsiveness. Male meadow voles (Microtus pennsylvanicus) of the two extreme phenotypes — reproductively photoresponsive and non-responsive — were either provided with a running wheel or housed without one. After 4 weeks with a wheel, the responsive voles had recovered full reproductive function, while the reproductive axes of responsive voles housed without wheels remained suppressed. Three experiments queried whether the use of a wheel would have reproductively stimulative effects in other rodents. First, intact male mice given access to wheels showed no increase in testis size when compared to mice housed without wheels. Likewise, locomotor activity had no effect on male rats whose testes were partially regressed in response to testosterone implants or on female mice whose estrous cycles were pheromonally suppressed by housing them in groups. Thus the neuroendocrine pathway used by locomotor activity to enhance the secretion of gonadotropin is specifically allied with the pathway used by photoperiod to control GnRH secretion.Abbreviations GnRH gonadotropin-releasing hormone - LH luteinizing hormone     

Responses to Environmental Enrichment Differ with Sex and Genotype in a Transgenic Mouse Model of Huntington's Disease

Background

Environmental enrichment (EE) in laboratory animals improves neurological function and motor/cognitive performance, and is proposed as a strategy for treating neurodegenerative diseases. EE has been investigated in the R6/2 mouse model of Huntington''s disease (HD), where increased social interaction, sensory stimulation, exploration, and physical activity improved survival. We have also shown previously that HD patients and R6/2 mice have disrupted circadian rhythms, treatment of which may improve cognition, general health, and survival.

Methodology/Principal Findings

We examined the effects of EE on the behavioral phenotype and circadian activity of R6/2 mice. Our mice are typically housed in an “enriched” environment, so the EE that the mice received was in addition to these enhanced housing conditions. Mice were either kept in their home cages or exposed daily to the EE (a large playground box containing running wheels and other toys). The “home cage” and “playground” groups were subdivided into “handling” (stimulated throughout the experimental period) and “no-handling” groups. All mice were assessed for survival, body weight, and cognitive performance in the Morris water maze (MWM). Mice in the playground groups were more active throughout the enrichment period than home cage mice. Furthermore, R6/2 mice in the EE/no-handling groups had better survival than those in the home cage/no-handling groups. Sex differences were seen in response to EE. Handling was detrimental to R6/2 female mice, but EE increased the body weight of male R6/2 and WT mice in the handling group. EE combined with handling significantly improved MWM performance in female, but not male, R6/2 mice.

Conclusions/Significance

We show that even when mice are living in an enriched home cage, further EE had beneficial effects. However, the improvements in cognition and survival vary with sex and genotype. These results indicate that EE may improve the quality of life of HD patients, but we suggest that EE as a therapy should be tailored to individuals.     

Response,use and habituation to a mouse house in C57BL/6J and BALB/c mice

Animal welfare depends on the possibility to express species-specific behaviours and can be strongly compromised in socially and environmentally deprived conditions. Nesting materials and refuges are very important resources to express these behaviours and should be considered as housing supplementation items. We evaluated the effects of one item of housing supplementation in standard settings in laboratory mice. C57BL/6JOlaHsd (B6) and BALB/cOlaHsd (BALB) young male and female mice, upon arrival, were housed in groups of four in standard laboratory cages and after 10 days of acclimatization, a red transparent plastic triangular-shaped Mouse House™ was introduced into half of the home cages. Animals with or without a mouse house were observed in various contexts for more than one month. Body weight gain and food intake, home cage behaviours, emotionality and response to standard cage changing procedures were evaluated. The presence of a mouse house in the home cage did not interfere with main developmental and behavioural parameters or emotionality of BALB and B6 male and female mice compared with controls. Both strains habituated to the mouse house in about a week, but made use of it differently, with BALB mice using the house more than the B6 strain. Our results suggest that mice habituated to the mouse house rather quickly without disrupting their home cage activities. Scientists can thus be encouraged to use mouse houses, also in view of the implementation of the EU Directive (2010/63/EU).     

Effects of cage density on behavior in young adult mice

Optimal housing conditions for mice can be achieved by minimizing environmental variables, such as those that may contribute to anxiety-like behavior. This study evaluated the effects of cage size on juvenile mice through assessment of differences in weaning weight, locomotor skills, and anxiety-like behavior. Eighteen pairs of male and pregnant female Swiss-Webster (Cr:SW) mice were housed in 3 different caging scenarios, providing 429, 505, or 729 cm2 of space. Litters were standardized to 10 pups per litter in each cage. Mice reared in each caging scenario were assessed with the open-field, light-dark exploration, and elevated plus-maze tests. No differences in weaning weight were noted. Mice reared in the 505- and 729-cm2 cages explored a significantly larger area of the open-field arena than did those in the 429-cm2 cages. Those reared in the 505-cm2 cages spent more time in the center of the open field than did those in the 729-cm2 cages, suggesting that anxiety-like behavior may be increased in the animals housed in the larger cages. This study did not establish a consistent link between decreased floor space and increased anxiety-like behavior; neither does there appear to be a consistent effect of available floor area on the development of locomotor skills on mouse pups.     

Response of Sod-2 enzyme activity to selection for high voluntary wheel running

The objective of this study was to examine the correlated response of anti-oxidant enzyme activity to selective breeding for increased voluntary wheel running in house mice. Activity of liver superoxide dismutase-2 (Sod-2), a free radical scavenger, was measured in four groups of mice. 'Active' individuals were housed in cages with attached wheels for 8 weeks beginning at weaning; 'sedentary' individuals were housed in cages with attached wheels that were prevented from rotating. Both of these treatments were applied to male and female mice from generation 14 of a replicated artificial selection experiment, which is composed of four lines selected for high wheel running and four randomly bred lines that serve as controls. In females, Sod-2 activity was significantly lower in selected vs control animals, regardless of presence/absence of a free-turning wheel. This difference suggests a trade-off between early-age voluntary wheel-running activity and Sod-2 activity. In males, Sod-2 activity was significantly affected by an interaction between selection group and activity group, with males from selected lines having lower Sod-2 activity relative to control males only in the sedentary treatment. These negative correlated responses of Sod-2 activity to selection on wheel running are discussed in the context of antagonistic pleiotropy models of aging and with respect to potential effects on lifespan.     

Housing of growing rabbits in individual,bicellular and collective cages: fear level and behavioural patterns

During growth (27 to 75 days of age), a total of 384 rabbits were kept in 72 individual cages, 48 bicellular cages (2 rabbits/cage) and 24 collective cages (9 rabbits/cage). To evaluate the effects of the housing system on the fear level and behavioural patterns of rabbits at the two ages (39 to 45 days and 66 to 73 days), a tonic immobility test and an open-field test were conducted and their behaviour was video recorded. In the tonic immobility test, the number of attempts to induce immobility (1.38) was lower, and the duration of immobility (47.8 s) was higher (0.05 < P < 0.01) in the rabbits housed in individual cages than in those kept in bicellular (1.72 attempts and 25.0 s of immobility) and collective cages (1.99 attempts and 25.0 s of immobility). During the open-field test, the rabbits from individual and bicellular cages showed higher latency (38.8 and 40.3 v. 27.0 s), a lower number of total (73.3 and 81.7 v. 91.9) and central displacements (3.6 and 2.8 v. 5.4) and a shorter running time (11.8 and 13.6 s v. 17.7 s) and the time biting the pen (5.5 and 9.1 s v. 28.2 s) compared with the rabbits kept in collective cages (0.05 < P < 0.001). During the 24-h video recording, the rabbits in individual and bicellular cages spent less time allogrooming (0.34% and 0.19% v. 1.44%), moving (0.74% and 0.60% v. 1.32%) and running (0.08% and 0.03% v. 0.21%) than the rabbits in the collective cages (0.01 < P < 0.001). The lowest numbers of alerts and hops were observed in the rabbits kept in bicellular cages. With increasing age, a lower number of rabbits were sensitive to the immobility test and more rabbits entered the pen spontaneously during the open-field test (P < 0.001). In conclusion, the rabbits in individual cages exhibited the highest fear level and incomplete behavioural patterns; the rabbits housed in collective cages showed the lowest fear levels and had the possibility of expressing a wider range of behaviour; and the rabbits in bicellular cages exhibited an inconsistent pattern of fear in the tonic immobility and open-field tests. Probably, these rabbits were in a less stressful condition compared with animals in individual cages because social contacts were allowed, even if freedom of movement was more limited.     

Effects of early weaning and housing conditions on the development of stereotypies in farmed mink

Female mink pups were weaned at 6, 8 or 10 weeks of age and subjected to two different housing conditions. They were either kept together with a single male sibling in traditional mink cages (30x45x90 cm) or housed socially with all litter-mates in an alternative system consisting of three adjoining traditional cages (90x45x90 cm). All cages were supplied with nest boxes. At 5 months of age, the siblings were removed leaving the females socially isolated in the two different cage systems. Females' stereotypies were quantified by repeated scanning observations under the social housing conditions immediately before removal of the siblings, and again at the age of 7 and 9 months, when the animals had stayed solitary in the two systems for 2 and 4 months. Solitary females showed significantly more stereotypies than females under social housing conditions in both cage systems. Stereotypies were more frequent in the smaller traditional cages. Stereotypies declined from 7 to 9 months of age among solitary animals in traditional cages but not in alternative cages. Early-weaned solitary females in traditional cages showed more stereotypies than later-weaned animals, but only when measured at the age of 7 months. It is suggested that early weaning, individual housing and small cages promote the development of stereotypies in farmed mink. The influence of early weaning on stereotypies seems to decline with age, while effects related to individual housing and small cages appear to be more persistent.     

The effect of housing and environmental enrichment on stereotyped behavior of adult vervet monkeys (Chlorocebus aethiops)

Little information is available on the response of vervet monkeys to different housing conditions or on the suitability of enrichment devices or methods for vervet monkeys. In this study, the authors evaluated the occurrence of stereotyped behavior in adult vervet monkeys under various conditions of housing and enrichment. The variables included cage size, cage level (upper or lower), enrichment with a foraging log, enrichment with an exercise cage and presence of a mate. The authors first determined the incidence of stereotyped behavior in captive-bred, singly housed adult female and male vervet monkeys. They then exposed monkeys to different housing and enrichment situations and compared the incidence of stereotyped behavior among the monkeys. The authors found that more females than males engaged in stereotyped behavior and that females, on average, engaged in such behavior for longer periods of time than males. Stereotyped behavior was most often associated with a small, single cage. The average amount of observed stereotyped activity in monkeys housed in a small cage was significantly lower when the monkeys had access to either a foraging log or an exercise cage. Stereotyped behavior was also lower in female monkeys that were housed (either with a male or without a male) in a larger cage. The least amount of abnormal behavior was associated with the largest, most complex and enriched housing situation. Males and females housed in cages on the lower level of two-level housing engaged in more stereotyped behavior than did monkeys housed in the upper level, regardless of the presence or type of enrichment provided.     

Effects of housing density on nasal pathology of breeding mice housed in individually ventilated cages

The 2011 edition of the Guide for the Care and Use of Laboratory Animals includes new recommendations for the amount of floor space that should be provided to breeding mice. When pairs or trios of continuously breeding mice are housed in shoebox cages, they may have less than this recommended amount of floor space. High housing densities may adversely affect animal health, for example, by compromising air quality inside the cage. Hence, some institutions are carefully reevaluating the microenvironments of breeding cages. The use of individually ventilated cages (IVCs) to house research mice allows for greater control over the quality of the cage microenvironment. The authors evaluated the microenvironments of shoebox cages in an IVC rack system housing breeding and non-breeding Swiss Webster mice. Ammonia concentrations were significantly higher in cages housing breeding trios with two litters. Histopathologic lesions attributable to inhaled irritants such as ammonia were found in mice housed in breeding pairs and trios. The authors conclude that the microenvironments of cages in an IVC rack system housing breeding pairs and trios may be detrimental to animal health.     

Preferences of mice, Mus musculus, for different types of running wheel

Mice are increasingly used in research. In particular, their wheel running is often used as a measure of activity, and as a marker of phase of circadian rhythms. Learning about the preferences of mice for different types of wheel may improve their welfare and suggest ways of increasing activity levels. Mice, Mus musculus, were given a choice between different types of running wheel by putting them in cages equipped with two wheels. Strong preferences were shown for wheels with a plastic mesh flooring, rather than the standard metal rods only. The mesh was even preferred over a solid base, although this effect was not seen in mice that had been given access only to wheels with the solid base immediately prior to the choice test. Small diameter wheels, sometimes sold as mouse wheels, were preferred less than standard-sized wheels with rods. The results suggest that types of running wheel often used in laboratories can be improved by considering the animals' preferences. The types of wheel tested here are easy to maintain and entail little additional cost, while increasing the mouse's interest in running and exercise.     

Close Proximity of the Heterosexual Partner Reduces the Physiological and Behavioral Consequences of Novel-Cage Housing in Black Tufted-Ear Marmosets (Callithrix kuhli)

The present studies assessed the extent to which heterosexual pairmates could buffer marmosets (Wied's black tufted-ear marmoset,Callithrix kuhli)against stress. Six male and six female marmosets from established groups were exposed to two experimental manipulations together with a control condition. Each condition lasted a total of 4 days. For the two experimental conditions, animals were removed from the family group and housed in a novel cage for 48 h in either the presence or the absence of the heterosexual pairmate. During the 48-h novel-cage housing period and for 48 h upon reunion of the subjects with the family group, concentrations of urinary cortisol were measured in the first void sample of the day and behavioral observations were conducted. When animals were housed alone in a novel cage they exhibited significant elevations in levels of urinary cortisol after 24 and 48 h of novel-cage exposure. In contrast, when marmosets were housed in the novel cage in the presence of the pairmate, levels of urinary cortisol did not change across the 4-day period. The presence of the social partner also reduced the behavioral manifestations of exposure to novelty. Upon reunion with the family group, animals that had been housed in the novel cage alone spent significantly more time in close proximity to the pairmate than animals that had been housed with the partner. A second experiment was conducted to determine the effect that separation from the pairmate, only (independent of any effects of novelty), had on levels of cortisol. Concentrations of urinary cortisol were measured in subjects housed in the familiar home cage, but in the absence of the pairmate, over a 48-h period and compared to concentrations of excreted cortisol immediately prior to separation. Separation from the pairmate did not elevate cortisol levels when the subject was housed in the home cage, suggesting that elevated cortisol levels in animals housed alone in the novel cage were in response to novelty exposure rather than to separation from the pairmate. Since the physical presence of the heterosexual partner reduced the physiological and behavioral effects of novel-cage housing, social attachments might function as homeostatic regulators of HPA function in marmosets.     

Anxiety-related defensive behavioral responses in mice selectively bred for High and Low Activity

High and Low Activity strains of mice (displaying low and high anxiety-like behavior, respectively) with 7.8–20 fold differences in open-field activity were selected and subsequently inbred to use as a genetic model for studying anxiety-like behavior in mice (DeFries et al., 1978, Behavior Genetics, 8:3-13). These strains exhibited differences in other anxiety-related behaviors as assessed using the light–dark box, elevated plus-maze, mirror chamber, and elevated square-maze tests (Henderson et al., 2004, Behavior Genetics, 34: 267-293). The purpose of these experiments was three-fold. First, we repeated a 6-day behavioral battery using updated equipment and software to confirm the extreme differences in anxiety-like behaviors. Second, we tested novel object exploration, a measure of anxiety-like behavior that does not rely heavily on locomotion. Third, we conducted a home cage wheel running experiment to determine whether these strains differ in locomotor activity in a familiar, home cage environment. Our behavioral test battery confirmed extreme differences in multiple measures of anxiety-like behaviors. Furthermore, the novel object test demonstrated that the High Activity mice exhibited decreased anxiety-like behaviors (increased nose pokes) compared to Low Activity mice. Finally, male Low Activity mice ran nearly twice as far each day on running wheels compared to High Activity mice, while female High and Low Activity mice did not differ in wheel running. These results support the idea that the behavioral differences between High and Low Activity mice are likely to be due to anxiety-related factors and not simply generalized differences in locomotor activity.     

The thalamic intergeniculate leaflet mediates locomotor activity-induced reversal of phenotype in photoperiod nonresponsive Siberian hamsters

The role of the intergeniculate leaflet of the thalamus (IGL) in photoperiod responsiveness was examined in a laboratory-selected line of photoperiod nonresponsive (NR) Siberian hamsters. NR hamsters fail to exhibit typical winter-type responses (i.e., gonadal regression and development of winter-type pelage) when exposed to short day lengths (e.g., 10 h of light/day). Earlier studies revealed that NR hamsters will exhibit winter-type responses when exposed to short photoperiod if they are given free access to a running wheel. The present study tested the hypothesis that this locomotor activity-induced reversal of phenotype is dependent on the IGL. Male NR hamsters underwent destruction of the IGL prior to being housed in short day lengths in cages equipped with running wheels. Activity rhythms were monitored for 8 weeks, after which time pelage response and paired testes weights were obtained. In contrast to sham-operated NR animals given access to running wheels, IGL-ablated animals showed no increase in the duration of nocturnal running wheel activity and became active later in the night than sham-lesioned animals. Lesioned animals also failed to exhibit the typical short photoperiod-induced gonadal regression and pelage molt. The results implicate the IGL in the mechanism by which running wheel activity can influence photoperiodic responses.     

Running wheel choice by Syrian hamsters

The present study investigated the preference of male and female Syrian hamsters, Mesocricetus auratus, for different types of running wheels. Hamsters were placed individually in sets of multiple cages linked by tunnels, each cage with a different running wheel. The number of wheel revolutions in each cage was tallied daily over 40 days. The hamsters did not express a preference when offered a choice of a running surface made of metal rods spaced 9 mm apart and a similar running surface covered in plastic mesh to prevent the possible slippage of feet between the rods. The hamsters did express a clear preference for larger wheels (35 versus 23 cm diameter), and for completely circular wheels over truncated ones. They neither favoured nor rejected wheels with small obstacles along the running surface. In all experiments, preferences were more strongly expressed by males than by females. Running wheels for hamsters may be improved by enlarging their diameter (to the standards often used for rats, if practically possible) and by ensuring good footing on the running surface (a space no larger than 9 mm between evenly spaced rods seems sufficient to achieve this, at least in large wheels and for hamsters older than 55 days). Installing obstacles along the running surface does not appear to make the wheel more interesting to hamsters.     

Scrounging facilitates social learning in common marmosets, Callithrix jacchus

The minimalistic environment of standard laboratory cages can adversely influence the responses of animals in standard behavioural tests and other aspects of the animals' biology. To avoid this, cages should provide for the animals' species-specific behavioural characteristics. We hypothesized that, given their possible capacity for colour vision, laboratory mice, Mus musculus, would show preferences between cages of different colours. Studies show that environmental colour can influence emotionality and task performance in humans, suggesting that cage colour could also affect emotionality and performance of mice in behavioural tests. Seventy-two mice were housed in home cages painted red, black, green or white. Five weeks later, 24 mice were placed individually into an apparatus allowing them to choose between cages of each of the home cage colours. Each mouse showed a highly significant preference, which overall, was unrelated to home cage colour. White cages were most preferred and red were least. Home cage colour had a significant effect on body weight and food consumption as well as on behaviour in a raised plus maze. Mice from red home cages spent most time in the closed arms, indicating greater anxiety, possibly suggesting that the reduced occupancy of the red preference cages resulted from avoidance of environmental conditions that induced a negative mental state. These findings show that laboratory mice have strong preferences between cages of different colours. We also found that an apparently inconsequential environmental variable, home cage colour, can influence responses in standard behavioural tests, which should be considered in assessing the external validity of such tests. Copyright 2003 The Association for the Study of Animal Behaviour. Published by Elsevier Ltd. All rights reserved.     

Social context affects the motivation of laboratory mice, Mus musculus, to gain access to resources

Consumer demand studies measuring the motivation that animals have for resources have almost exclusively used gregarious species, but have tested animals housed in isolation. The motivation that gregarious animals have for resources could be increased or decreased by the presence or absence of companion animals, that is, the social context. I trained a single mouse from each of six groups to perform an operant task that was quantifiable and would not be performed fortuitously or learnt by nontrained mice. The trained mice were housed with nontrained cagemates in a start cage that required them to perform the operant task to enter a resource cage providing either additional space or a running wheel. The presence of cagemates in the start cage significantly decreased the number of visits/24 h to the running wheel, the number of bouts and the duration of wheel running, but had no effect on the number of visits to additional space. The presence of cagemates increased the time spent in the resource cage whether this provided additional space or a running wheel, indicating that the mice overcompensated for the reduced frequency of visits. These results show that social context can have a significant effect on the motivation that animals have for resources, and that this effect is resource dependent. The influence of social context should therefore be considered in the design and interpretation of consumer demand studies. Copyright 2003 Published by Elsevier Ltd on behalf of The Association for the Study of Animal Behaviour.