The motivation of group-housed laboratory mice, Mus musculus, for additional space

The limited amount of space that we provide for research animals can adversely influence the validity of behavioural data and animal welfare. Consumer demand studies have examined the strength of motivation that animals have for additional space; however, one problem of these studies is that the animals tested were generally of a gregarious species, but were tested in isolation. I trained a single mouse from each of six groups to perform an operant task to gain access to an additional 319, 777, or 1600 cm² while group-housed. As the cost of visits increased, the mice continued to gain access to the additional space, although the numbers of visits and the time in the additional space decreased. The slopes of the demand functions for the three amounts of additional space ranged between 0.64 and 0.44, indicating that the mice perceived additional space to be an important resource. The slopes of the demand functions, the areas under the demand functions and the Y-axis intercepts were not significantly different between the three sizes of additional space, thus corroborating previous similar work. These results indicate that group-housed laboratory mice in standard laboratory cages were highly motivated for additional space, but did not discriminate between the amounts offered.     

Behavioral assessment of intermittent wheel running and individual housing in mice in the laboratory

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.     

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.     

Using the Activity-based Anorexia Rodent Model to Study the Neurobiological Basis of Anorexia Nervosa

Anorexia nervosa (AN) is a psychiatric illness characterized by excessively restricted caloric intake and abnormally high levels of physical activity. A challenging illness to treat, due to the lack of understanding of the underlying neurobiology, AN has the highest mortality rate among psychiatric illnesses. To address this need, neuroscientists are using an animal model to study how neural circuits may contribute toward vulnerability to AN and may be affected by AN. Activity-based anorexia (ABA) is a bio-behavioral phenomenon described in rodents that models the key symptoms of anorexia nervosa. When rodents with free access to voluntary exercise on a running wheel experience food restriction, they become hyperactive – running more than animals with free access to food. Here, we describe the procedures by which ABA is induced in adolescent female C57BL/6 mice. On postnatal day 36 (P36), the animal is housed with access to voluntary exercise on a running wheel. After 4 days of acclimation to the running wheel, on P40, all food is removed from the cage. For the next 3 days, food is returned to the cage (allowing animals free food access) for 2 hr daily. After the fourth day of food restriction, free access to food is returned and the running wheel is removed from the cage to allow the animals to recover. Continuous multi-day analysis of running wheel activity shows that mice become hyperactive within 24 hr following the onset of food restriction. The mice run even during the limited time during which they have access to food. Additionally, the circadian pattern of wheel running becomes disrupted by the experience of food restriction. We have been able to correlate neurobiological changes with various aspects of the animals’ wheel running behavior to implicate particular brain regions and neurochemical changes with resilience and vulnerability to food-restriction induced hyperactivity.     

The reinforcing property and the rewarding aftereffect of wheel running in rats: a combination of two paradigms

Wheel running reinforces the behavior that generates it and produces a preference for the context that follows it. The goal of the present study was to demonstrate both of these effects in the same animals. Twelve male Wistar rats were first exposed to a fixed-interval 30 s schedule of wheel-running reinforcement. The operant was lever-pressing and the reinforcer was the opportunity to run for 45 s. Following this phase, the method of place conditioning was used to test for a rewarding aftereffect following operant sessions. On alternating days, half the rats responded for wheel-running reinforcement while the other half remained in their home cage. Upon completion of the wheel-running reinforcement sessions, rats that ran and rats that remained in their home cages were placed into a chamber of a conditioned place preference (CPP) apparatus for 30 min. Each animal received six pairings of a distinctive context with wheel running and six pairings of a different context with their home cage. On the test day, animals were free to move between the chambers for 10 min. Results showed a conditioned place preference for the context associated with wheel running; however, time spent in the context associated with running was not related to wheel-running rate, lever-pressing rate, or post-reinforcement pause duration.     

Effects of long-term physical exercise on skeletal muscles in senescence-accelerated mice (SAMP8)

We examined the effect of long-term exercise on the prevention of sarcopenia using a senescence-accelerated-prone mice (SAMP8) model. Mice were housed in a wheel cage for 25 weeks to increase voluntary exercise. At week 23, endurance running capacity was examined using a treadmill. In a treadmill running test, the wheel cage group had increased endurance running capacity, which suggests that aging-related loss of muscle function was recovered by long-term exercise. Mice were sacrificed and microarray analysis revealed that genes involved in protein synthesis and degradation were upregulated in the skeletal muscles of the wheel cage group, suggesting accelerated protein turnover. Total body and adipose tissue weights decreased following the use of the wheel cage. Thus, long-term, spontaneous physical exercise may assist in recovering from aging-related sarcopenia (loss of muscle function) and obesity.     

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.     

Effect of cage enrichment on the daily use of running wheels by Syrian hamsters

Institutional animal care committees may one day require for the welfare of captive hamsters more floor space and the introduction of tunnels and toys. As hamsters are popular animal subjects in chronobiological research, and as clock phase is usually measured through running wheel activity, it is important to determine what effect cage enrichment might have on daily wheel use. Here the daily number of wheel revolutions, the daily duration of the running activity phase, the phase relationship between lights-off and onset of running activity, and the free-running period of circadian activity rhythms were measured in Syrian hamsters, Mesocricetus auratus, housed in single cages or in multiple cages linked by tunnels and supplied with commercial wooden toys. Free-running periodicity was not affected by cage enrichment. In multiple-cage systems, there were fewer daily revolutions, shorter wheel-running activity phases, and delayed running activity onsets. These effects, however, were small as compared to interindividual and week-to-week variation. They were statistically significant only under a light:dark cycle, not in constant darkness, and only when interindividual variation was eliminated through a paired design or when the number of cages was increased to five (the maximum tested). Daily wheel use is thus affected by cage enrichment, but only slightly.     

Colour learning in two behavioural contexts: how much can a butterfly keep in mind?

Here we examine the ability of butterflies to learn colour cues in two different behavioural contexts, nectar foraging and oviposition, more or less simultaneously. We first trained female Battus philenor (Papilionidae) butterflies to associate a given colour with the presence of host plant leaf extract and assayed their colour preference; we then trained a subset of these butterflies to associate a second colour with the presence of sucrose solution and assayed colour preference once more. When offered an array of four unscented and unrewarding coloured models, ‘single-trained’ butterflies consistently alighted most frequently on their oviposition training colour. Green-trained butterflies landed on nontrained colours only about 4% of the time, while butterflies trained to red, yellow or blue made about 23% of their landings on nontrained colours; of those nontrained landings, most were on green. The majority of ‘dual-trained’ butterflies made the greatest number of visits to both training colours in the appropriate behavioural context; that is, they probed the models of their sucrose-associated colour and alighted on the models of their oviposition-associated colour. Landings or probes on nontrained colours in one context were consistently biased towards what was learned in the alternative context, suggesting an information-processing constraint in the butterflies. This paper provides a clear demonstration that butterflies can learn in two behavioural contexts within a short span of time. A capacity for such dual conditioning presumably permits female butterflies to forage effectively for egg-laying sites and nectar resources even when those activities are intermingled in time. Copyright 2003 Published by Elsevier Science Ltd on behalf of The Association for the Study of Animal Behaviour.       

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     

Papillary mechanics and cardiac morphology of infarcted rat hearts after training

Infarction of the left ventricle was induced by ligation of the coronary artery in male Sprague-Dawley rats under ketamine-xylazine anesthesia. Three weeks after surgery, animals were assigned to a trained (n = 21; running at 20 m/min, 10% grade, 1 h/day, 5 days/wk) or nontrained group (n = 23) for an additional 8 wk. A third, sham-operated control group (n = 16) remained cage sedentary for 11 wk. Ventricular mass was greater in the trained and nontrained infarct groups [1,335 +/- 57.3 and 1,414 +/- 56.1 mg, respectively (mean +/- SE)] compared with the control group (1,155 +/- 50.9 mg) (P less than or equal to 0.05). The diameter of septal fibers was 13% greater in the trained and 17% greater in the nontrained infarct groups compared with control. The specific peak developed force and maximum rate of force development of left ventricular papillary muscle in vitro were 75 and 62% greater in both infarcted groups compared with the control group; these variables were unaffected by training. Myofibrillar adenosine triphosphatase activity of septum was 20% lower in both infarct groups compared with sham-operated animals. We conclude that exercise training did not alter the magnitude of morphological and physiological adaptations to infarction.     

Effect of access to a running wheel on behavior of C57BL/6J mice.

BACKGROUND AND PURPOSE: To evaluate how and when mice run on a running wheel and how ad libitum access to the wheel affect behavior, feed intake, and weight gain. METHODS: Seventeen 2-month-old C57BL/6J mice had access to the wheel, whereas 19 control mice did not. After 3 to 6.5 weeks, behavior was video-recorded over 24 h for each mouse. RESULTS: Experimental mice ran an average 2 km/24 h in 114 min. Highest running activity took place at the onset of darkness. Experimental mice spent 22 min more feeding on the cage floor than did control mice. These times were deducted from those for all other behaviors: 74 min from resting time, 39 min from climbing and feeding on the cage lid, 14 min from locomotion on the cage floor, and 10 min from grooming. In relative figures, deduction from sleeping time was only 9%, whereas climbing time was halved. CONCLUSIONS: Climbing on the cage lid has a similar circadian rhythm as does wheel running and high-energy expenditure. Because experimental mice climbed less, their weight gain and feed intake were similar to those of control mice. Thus, wheel running can substitute for other forms of energy-consuming behaviors and vice versa.     

Voluntary exercise delays monogenetic obesity and overcomes reproductive dysfunction of the melanocortin-4 receptor knockout mouse

The melanocortin system is involved in hypothalamic regulation of energy homeostasis. The melanocortin-4 receptor (MC4R) has been linked to both obesity and reproductive dysfunction. Deletion of the MC4R from the mouse genome has resulted in phenotypes including adult onset obesity, hyperphagia, and difficulty in reproducing when homozygote parents are bred. Additionally, polymorphisms of the human MC4R have been identified in morbidly obese children and adults. Herein, we have identified that voluntary exercise, provided via the presence of a running wheel, impedes the monogenetic obesity (at 20 weeks of age running wheel housed body weight=31+/-1.8 g versus conventionally housed body weight=41+/-2.3 g, a 25% decrease in body weight p<0.01), hyperphagia (average cumulative food intake is not statistically different than wild type mice housed in running wheel cages), and reproductive dysfunction phenotypes associated with the MC4R knockout mice housed by conventional means. These data demonstrate the novel finding that voluntary exercise at a young age may hinder genetically induced obesity.     

Testing resource value in group-housed animals: an investigation of cage height preference in laying hens

To avoid unpredictable social effects, animals' behavioural priorities are almost always tested using individuals housed singly, yet many species kept commercially are social animals housed in groups. Our aim was to develop a method of investigating environmental preference in group-housed laying hens, Gallus gallus domesticus, that maximised the external validity of our findings. In a simple test of preference, eight groups of ten hens were given free choice between furnished cages with minimum heights of 38 cm (low) and 45 cm (high). A preference for one cage height over the other would be evident as a shift from a binomial distribution of flock sizes in the two cages. No height preference was found as hens distributed evenly between the two cages more frequently than was expected. This suggests at high stocking densities maximising average inter-individual distance could be a priority over increased cage height. In a second experiment, to investigate the value that hens placed on a change in cage height; a 'cost' in the form of a narrow gap was imposed on movement from a low or high start cage to a high or low target cage, respectively. Cage height did not influence the latency of the first three hens to enter the target cage. However, latencies for subsequent hens were shorter and more hens worked to access a high target cage than a low target cage. We suggest that titrating animals' willingness to tolerate higher stocking densities against access to a resource could be an effective way to compare responses of group-housed animals to resources that are expected to satisfy the same motivational state.     

If you knew what was good for you! The value of environmental enrichments with known welfare benefits is not demonstrated by sows using operant techniques

This study assessed the motivation of gestating sows housed in standard, barren gestation stalls (used for breeding/implantation and/or gestation) for access to environmental enrichment. Enrichment consisted of a cotton rope or rubber mat in comparison to positive (additional food when fed at commercial levels) and negative (empty trough) controls. Although environmental enrichment may improve animal welfare, sows' valuation of enrichments is largely unknown. This study used an operant panel and obtained behavioral measures to quantify motivation. As indicated by a higher price paid and lower latencies to press the panel and enter the treatment stall (all comparisons, p < .05), sows demonstrated higher motivation for food compared with all treatments. Sows housed in gestation stalls did not demonstrate high motivation via operant responding for a cotton rope or a rubber mat; nor did they demonstrate any differences in behavioral measures (all comparisons, p > .10). Although sows' motivation for a mat did not differ from that for an empty trough, previous work has demonstrated the welfare benefits associated with comfort flooring.     

Effects of early‐life exposure to Western diet and wheel access on metabolic syndrome profiles in mice bred for high voluntary exercise

Experimental studies manipulating diet and exercise have shown varying effects on metabolic syndrome components in both humans and rodents. To examine the potential interactive effects of diet, exercise and genetic background, we studied mice from four replicate lines bred (52 generations) for high voluntary wheel running (HR lines) and four unselected control lines (C). At weaning, animals were housed for 60 days with or without wheels and fed either a standard chow or Western diet (WD, 42% kcal from fat). Four serial (three juvenile and one adult) blood samples were taken to measure fasting total cholesterol (TC), high‐density lipoprotein cholesterol (HDL‐C), triglycerides and glucose. Western diet was obesogenic for all mice, even after accounting for the amount of wheel running and kilojoules consumed. Western diet significantly raised glucose as well as TC and HDL‐C concentrations. At the level of individual variation (repeatability), there was a modest correlation (r = 0.3–0.5) of blood lipids over time, which was reduced with wheel access and/or WD. Neither genetic selection history nor wheel access had a statistically significant effect on blood lipids. However, HR and C mice had divergent ontogenetic trajectories for body mass and caloric intake. HR mice also had lower adiposity, an effect that was dependent on wheel access. The environmental factors of diet and wheel access had pronounced effects on body mass, food consumption and fasting glucose concentrations, interacting with each other and/or with genetic strain. These data underscore the importance (and often unpredictable nature) of genotype‐by‐environment and environment‐by‐environment interactions when studying body weight regulation.     

If You Knew What Was Good For You! The Value of Environmental Enrichments With Known Welfare Benefits Is Not Demonstrated by Sows Using Operant Techniques

This study assessed the motivation of gestating sows housed in standard, barren gestation stalls (used for breeding/implantation and/or gestation) for access to environmental enrichment. Enrichment consisted of a cotton rope or rubber mat in comparison to positive (additional food when fed at commercial levels) and negative (empty trough) controls. Although environmental enrichment may improve animal welfare, sows' valuation of enrichments is largely unknown. This study used an operant panel and obtained behavioral measures to quantify motivation. As indicated by a higher price paid and lower latencies to press the panel and enter the treatment stall (all comparisons, p < .05), sows demonstrated higher motivation for food compared with all treatments. Sows housed in gestation stalls did not demonstrate high motivation via operant responding for a cotton rope or a rubber mat; nor did they demonstrate any differences in behavioral measures (all comparisons, p > .10). Although sows' motivation for a mat did not differ from that for an empty trough, previous work has demonstrated the welfare benefits associated with comfort flooring.     

Voluntary exercise impairs initial delayed spatial alternation performance in estradiol treated ovariectomized middle-aged rats

Estrogens differentially modulate behavior in the adult female rodent. Voluntary exercise can also impact behavior, often reversing age associated decrements in memory processes. Our research group has published a series of papers reporting a deficit in the acquisition of an operant working memory task, delayed spatial alternation (DSA), following 17β-estradiol treatment to middle-aged ovariectomized (OVX) rats. The current study examined if voluntary exercise could attenuate the 17β-estradiol induced deficits on DSA performance. OVX 12-month old Long–Evans rats were implanted with a Silastic capsule containing 17β-estradiol (10% in cholesterol: low physiological range) or with a blank capsule. A subset of the 17β-estradiol and OVX untreated rats were given free access to a running wheel in their home cage. All rats were tested for 40 sessions on the DSA task. Surprisingly, we found running wheel access to impair initial acquisition of the DSA task in 17β-estradiol treated rats, an effect not seen in OVX untreated rats given running wheel access. This deficit was driven by an increase in perseverative responding on a lever no longer associated with reinforcement. We also report for the first time a 17β-estradiol induced impairment on the DSA task following a long intertrial delay (18-sec), an effect revealed following more extended testing than in our previous studies (15 additional sessions). Overall, running wheel access increased initial error rate on the DSA task in 17β-estradiol treated middle-aged OVX rats, and failed to prevent the 17β-estradiol induced deficits in performance of the operant DSA task in later testing sessions.     

Effects of housing density on Long Evans and Fischer 344 rats

At many breeding facilities, rats are housed at relatively high densities until they are 5 weeks old, at which point they are either shipped for research or rehoused at standard cage densities according to weight. The authors carried out a pilot study in Long Evans and in Fischer 344 rats to investigate whether continuing to house rats at high densities (24 in(2) floor space per rat) past the age of 5 weeks, through puberty and into adulthood would alter behavioral or physiological parameters compared with raising rats at standard densities (about 72 in(2) floor space per rat). After rats reached puberty, the authors rehoused them with unfamiliar cagemates. The researchers evaluated clinical and behavioral signs of stress, weight, blood glucose concentration, white blood cell count and serum corticosterone concentration. Overall, cage density had little effect on the parameters measured, though gender seemed to affect stress in Long Evans rats. The results suggest that rats of these strains can be raised at the higher densities tested until any age and regrouped with unfamiliar cagemates without compromising rats' welfare or subsequent experimental data.