Are the effects of different enrichment designs on the physiology and behaviour of DBA/2 mice consistent?

Environmental enrichment is intended to improve the well-being of laboratory animals. Although many researchers have indicated that environmental enrichment may enhance animal well-being, there is some evidence that enrichment differs in its effects on physiology and behaviour between species and strains. The present study focuses on the effects of different enrichment designs on the physiology and behaviour of male and female DBA/2 mice. A total of 48 DBA/2J mice, 24 males and 24 females were used for this experiment. Upon arrival at about 3 weeks of age, the animals were randomly allotted to three experimental groups: NE, non-enrichment; E1, enriched with nest box, wooden climbing bar and nest material according to Scharmann (1993); E2, enriched with horizontal and vertical dividers, modified from Haemisch and G?rtner (1994). Same-sex groups of four mice were housed for 12 weeks in type III Makrolon cages with (E1 or E2) or without (NE) enrichment objects. Behavioural performance (Open Field, Food Drive and Elevated Plus Maze tests) and physiological traits (haematological variables, body weight and organ weights, corticosterone and thyroxine levels) were measured. This study observed that enrichment had significant effects on the mean values of body weight (females), Open Field and Food Drive tests. The most significant housing differences were found between the E2 and NE/E1 groups. Furthermore, sex differences in the NE, E1 and E2 groups were not consistent for several variables (growth rate, relative weights of spleen, kidney and heart, Food Drive and Elevated Plus Maze behavioural performance). There was often a higher coefficient of variation (CV) in the E1 and E2 groups as compared to the NE group, chiefly in physiological traits and in the Open Field and Food Drive tests. The results of this study indicate, that the effects of enrichment designs used in the present study are not consistent, but vary according to sex and the variable studied.     

The Influence of the Location of a Nest Box in an Individually Ventilated Cage on the Preference of Mice to Use It

The improvement of housing conditions for mice by using environmental enrichment materials is of high concern for the scientific community. Plastic, autoclavable nest boxes are commercially available and ready to use for specific cases such as in individually ventilated cages, metabolic cages, or during toxicological studies. The aim of this study was to see if the location of the nest box within the cage could influence the mice to prefer and use it. Located on the cage floor or hung from the cage lid, a nest box (MPLEX, Otto Environmental, Milwaukee, Wisconsin), enriched the cages. The study concluded that the location of the nest boxes in the individually ventilated cage plays a significant role in the mice preferring to use it or to avoid it. It is also important to use environmental enrichment items that provide animals with the possibility of expressing their preferences and manipulating them in a way to cope better with their environmental conditions.     

The influence of the location of a nest box in an individually ventilated cage on the preference of mice to use it

The improvement of housing conditions for mice by using environmental enrichment materials is of high concern for the scientific community. Plastic, autoclavable nest boxes are commercially available and ready to use for specific cases such as in individually ventilated cages, metabolic cages, or during toxicological studies. The aim of this study was to see if the location of the nest box within the cage could influence the mice to prefer and use it. Located on the cage floor or hung from the cage lid, a nest box (MPLEX, Otto Environmental, Milwaukee, Wisconsin), enriched the cages. The study concluded that the location of the nest boxes in the individually ventilated cage plays a significant role in the mice preferring to use it or to avoid it. It is also important to use environmental enrichment items that provide animals with the possibility of expressing their preferences and manipulating them in a way to cope better with their environmental conditions.     

The effect of preweaning and postweaning housing on the behaviour of the laboratory mouse (Mus musculus)

Standard housing for laboratory mice severely restricts natural behaviour and the control that the animal has over its environment. Providing the cage with objects is a method that has been used to both increase environmental complexity, promote the performance of natural behaviour and provide greater controllability for the animal. This method of furnishing cages has mostly been studied in adult animals, and little is known about the influence that the preweaning environment has on the behaviour of mice as adults. This study aimed to investigate the effects on mice behaviour of preweaning and postweaning housing environment. In this experiment, 64 pairs of animals of the strain C57BL/6J were used. Half of the animals were born and reared until weaning in standard cages and the other half in cages twice the size of the standard and furnished with nesting material, a cardboard tube, a PVC nest box and a wooden chewblock. After weaning, half the animals in each group were changed to the other type of cage, whereas the other half remained in the same environment; in both cases they were kept in single-sex pairs of littermates. Behaviour during the dark, active period was studied through video recordings. We found no main effects of preweaning environment on behaviour; however, mice moved from furnished to standard cages at weaning showed a decrease in inactive behaviour at four weeks of age. Mice housed after weaning in standard cages spent less time inactive, and more time engaging in activities like feeding and drinking, self-grooming and allogrooming. A sex difference was also found, in that females showed a greater performance of exploratory behaviour as well as a greater prevalence of stereotypies. The use of different objects and locations within the furnished cage was also analysed at both ages. Results show that at eight weeks of age mice spent more time at the top of the cage, and that the use of the nest box (although not for resting) increased between four and eight weeks. Mice were found to use the nest box as a nesting site/sleeping place only at age four weeks, whereas they always used the nesting material for sleeping.     

Effects of different forms of environmental enrichment on behavioral,endocrinological, and immunological parameters in male mice

This study investigated effects of different forms of environmental enrichment on behavioral, endocrinological, and immunological parameters in male mice. For this purpose, animals of the inbred strain CS were kept in groups of four males under three different housing conditions: (A) nonstructured Makrolon type III laboratory cages ("standard-housing" = S); (B) equivalent laboratory cages that were enriched with a box and a scaffolding ("enriched-housing" = E); and (C) spacious terraria that were structured richly ("super-enriched-housing" = SE). Both forms of enrichment caused a sharp rise in aggressive behavior, though play behavior was increased in E and SE mice, too. Levels of sociopositive behaviors in S and SE mice were higher than those in E mice. Plasma corticosterone concentrations and adrenal tyrosine hydroxylase activities were significantly increased in male mice kept in both forms of enriched cages, indicating an activation of the pituitary-adrenocortical and the adrenomedullary systems. The behavioral and endocrinological differences were partly reflected by immunological parameters: SE mice had levels of IgG1 and ratios of IFN-gamma/IL-10 and IL-2/IL-10 significantly lower than those of S mice. Ratios of IgG2a/IgG1 were significantly higher in SE mice. The absolute percentages of CD8 cells in E-mice were significantly lower than those in S mice. Despite the elevated levels of stress hormones under both forms of enriched housing, the behavioral parameters also indicate positive effects of the enrichment, especially on SE animals. Obviously, an environmental enrichment is beneficial for male mice as long as the spatial conditions are generous enough to allow coping with the increased aggression brought about by the enrichment.     

Environmental modification and agonistic behavior in NIH/S male mice: nesting material enhances fighting but shelters prevent it

Outbred NIH/S male mice were housed from weaning in groups of 4 without enrichment (control) or with nesting material (nest), nesting material and a box (nest-and-box), or nesting material and a tube (nest-and-tube) as environmental modification. The aim of the study was to investigate effects of widely recommended nesting material and additional shelters on male mice. The aggressiveness of the mice in their home cages clearly increased in the nest group, as assessed by the number of wounds. In the nest group, fighting was a stressful situation for the mice, leading to changes in weight gain and in the weights of the thymus, adrenals, spleen, and epididymal adipose tissue. Moreover, the agonistic behavior of these mice toward an intruder was increased both in individual tests (an intruder with the individual mouse) and group tests (an intruder with a group of mice). The provision of a box or tube as a shelter, in addition to nesting material, prevented intracage fighting and did not lead to alterations in the weight gain or organ weights of the mice. However, the agonistic behavior of mice with shelters was slightly increased in behavioral tests. Anxiety in the elevated plus-maze was not affected by any of the housing systems. In conclusion, the agonistic behavior of NIH/S mice, an aggressive strain, seemed to be easily enhanced by these environmental modifications. The suitability of any enrichment should be carefully evaluated, especially when highly aggressive mice are used.     

The effects of enriching laboratory cages using various physical structures on multiple measures of welfare in singly-housed rats

The single housing of laboratory rats may be recommended in some situations such as hypothesis-driven or test-specific studies, during electroencephalogram recording of phases of sleep and after surgical procedures. However, as single housing of laboratory rats has been shown to be stressful, modification of the housing environment is needed to improve the welfare of these animals. This experiment was carried out to investigate the long-term effects of environmental enrichment on some behavioural, physiological, pathological and psychological measures of welfare. With two batches of animals, 24 rats were housed singly in either enriched cages (EC) (n = 12 cages) or unenriched cages (UC) (n = 12 cages). Behaviour was sampled every week and so was body weight and weight gain over a six-week observation period. Behaviours of the rats in the elevated plus-maze were recorded on the seventh week, whereas organ weights were recorded postmortem. The results revealed that long-term single housing of rats in super-enriched cages increased levels of indicators of good welfare including sleep, exploration, movement and feeding behaviour, body weights, weight gains and the relative weights of the thymus gland and spleen, and decreased levels of indicators of poor welfare such as stationary behaviour and the relative weight of adrenal glands. Thus, enrichment of conventional cages of newly weaned singly-housed laboratory rats with multiple physical structures appeared to improve their ability to control the environment and to promote their species-specific behaviour; changes that can ultimately result in good welfare.     

Improving housing conditions for laboratory mice: a review of "environmental enrichment"

Laboratory animal facilities have been designed to provide a standard environment where animals can be kept in good physical health at the same time as economic and ergonomic considerations are met. Recognizing the potential welfare problem associated with behavioural restriction in such housing systems, a number of attempts have been made to improve this environment, generally described under the term "environmental enrichment". Modifications of cages for mice usually consist of providing material for nest building and structures which can serve as hiding places and/or for climbing. We have reviewed 40 studies carried out between 1987 and 2000, in which preferences as well as the effect of housing modifications have been studied. Mice will work for access to nesting material and make use of this material to make nests in which they rest. They prefer a more complex cage to the standard cage and will also work for access to cages with shelter and raised platforms. On the basis of present knowledge, it is recommended that mice should have access to nesting material. Strategies for future research are outlined in the article.     

Housing mice in the individually ventilated or open cages—Does it matter for behavioral phenotype?

Individually ventilated caging (IVC) systems for rodents are increasingly common in laboratory animal facilities. However, the impact of such substantial change in housing conditions on animal physiology and behavior is still debated. Most importantly, there arise the questions regarding reproducibility and comparison of previous or new phenotypes between the IVC and open cages. The present study was set up for detailed and systematic comparison of behavioral phenotypes in male and female mice of three widely used inbred strains (C57BL/6JRccHsd, DBA/2JRccHsd, 129S2/SvHSd) after being kept in two housing environments (IVC and open cages) for 6 weeks (since 4 weeks of age) before behavioral testing. The tests addressed exploratory, anxiety‐like and stress‐related behavior (light‐dark box, open field, forced swim test, stress‐induced hyperthermia), social approach and species‐specific behavior (nest building, marble burying). In all tests, large and expected strain differences were found. Somewhat surprisingly, the most striking effect of environment was found for basal body temperature and weight loss after one night of single housing in respective cages. In addition, the performance in light‐dark box and open field was affected by environment. Several parameters in different tests showed significant interaction between housing and genetic background. In summary, the IVC housing did not invalidate the well‐known differences between the mouse strains which have been established by previous studies. However, within the strains the results can be influenced by sex and housing system depending on the behavioral tasks applied. The bottom‐line is that the environmental conditions should be described explicitly in all publications.     

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.     

Effects of Increased Spatial Complexity on Behavioural Development and Task Performance in Lister Hooded Rats

Enhancing laboratory animal welfare, particularly in rodents, has been achieved through environmental enrichment in caging systems. Traditional enrichment such as adding objects has shown to impact development, reproductive and maternal performance as well as cognition. However, effects of increased spatial complexity as part of larger novel caging systems have not been investigated. While adoption of caging systems with increased spatial complexity seems uncontroversial from a welfare perspective, effects of such housing on the development and task performance of experimental animals remains unclear. In this study, we investigate differences in key behaviours and cognitive performance between Lister Hooded rats housed in traditional (single-shelf) cages (‘basic’) and those housed in larger cages with an additional shelf (‘enriched’). We found minor differences in maternal behaviour, such as nursing and offspring development. Further, we compared task performance in females, using a hippocampus-dependent task (T-maze) and a hippocampus-independent task (Novel Object Recognition, NOR). While in the T-maze no differences in either the rate of learning or probe trial performance were found, in the NOR task females housed in enriched cages performed better than those housed in basic cages. Our results show that increased spatial complexity does not significantly affect development and maternal performance but may enhance learning in females for a non-spatial task. Increased spatial complexity does not appear to have the same effects on behaviour and development as traditional enrichment. Thus, our results suggest no effect of housing conditions on the development of most behaviours in experimental animals housed in spatially enriched caging systems.     

Mandatory " enriched" housing of laboratory animals: the need for evidence-based evaluation

Environmental enrichment for laboratory animals has come to be viewed as a potential method for improving animal well-being in addition to its original sense as a paradigm for learning how experience molds the brain. It is suggested that the term housing supplementation better describes the wide range of alterations to laboratory animal housing that has been proposed or investigated. Changes in the environments of animals have important effects on brain structure, physiology, and behavior--including recovery from illness and injury--and on which genes are expressed in various organs. Studies are reviewed that show how the brain and other organs respond to environmental change. These data warrant caution that minor cage supplementation intended for improvement of animal well-being may alter important aspects of an animal's physiology and development in a manner not easily predicted from available research. Thus, various forms of housing supplementation, although utilized or even preferred by the animals, may not enhance laboratory animal well-being and may be detrimental to the research for which the laboratory animals are used.     

Computer-task testing of rhesus monkeys (Macaca mulatta) in the social milieu

Previous research has demonstrated that a behavior and performance testing paradigm, in which rhesus monkeys (Macaca mulatta) manipulate a joystick to respond to computer-generated stimuli, provides environmental enrichment and supports the psychological well-being of captive research animals. The present study was designed to determine whether computer-task activity would be affected by pair-housing animals that had previously been tested only in their single-animal home cages. No differences were observed in productivity or performance levels as a function of housing condition, even when the animals were required to “self-identify” prior to performing each trial. The data indicate that cognitive challenge and control are as preferred by the animals as social opportunities, and that, together with comfort/health considerations, each must be addressed for the assurance of psychological well-being.     

Behavioral effects of enrichment on singly-housed,yearling rhesus monkeys: An analysis including three enrichment conditions and a control group

Three cohorts of yearling rhesus monkeys (Macaca mulatta) were maintained in single cages for one year as part of a derivation program to produce a breeding colony of specific pathogen-free (SPF) monkeys. During this year of social restriction, subjects were provided with three different types of environmental enrichment (physical, feeding, and sensory) to counteract the known effects of social restriction and to quantify the effects of these different conditions of enhancement on their behavior. Focal animal observations were conducted on enriched and control subjects for all cohorts. Enrichment conditions were presented in a different order to each cohort. Monkeys provided with enrichment spent significantly more time playing and less time self-grooming than did control monkeys in unenriched cages, suggesting that the overall enrichment program was of some benefit to the monkeys, because these changes in behavior were in species-typical directions. Among enriched subjects only, there were significant differences in the amount of time spent drinking, grooming, feeding, playing, exploring, and using enrichment across the three enrichment conditions. Both the physical and feeding enrichment conditions led to species-appropriate changes in behavior, therefore enhancing psychological well-being as some define it. Sensory enrichment was of little benefit. The first cohort was housed indoors, received less stimulation from the environment outside of the single cage, and used enrichment more than did the other two cohorts housed outdoors. This suggests that the external environment influences behavior in the single cage and that enrichment may be most effective for animals housed indoors. © 1995 Wiley-Liss, Inc.     

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.     

The effect of daily disturbance on the breeding performance of mice

The United Kingdom Home Office Code of Practice for the housing and care of breeding animals requires that, 'the general well-being of all animals must be checked at least once daily'. However, excessive daily disturbance of rodent breeding colonies could be counter-productive to animal welfare if it increases pre-weaning mortality. An experiment involving 100 breeding cages of BALB/c mice compared daily inspection of the mouse cages, but without disturbing the mice within the nest, with daily inspection in which every individual was studied even if this involved removing the cage lid and disturbing the nest. No statistically significant differences were found between the two groups in breeding performance or pre-weaning mortality, though the disturbed group produced marginally fewer offspring and had slightly higher mortality. Average weaning weight did not differ between the groups, but sexual dimorphism at weaning was significantly increased in the disturbed group. It is concluded that there are unlikely to be any welfare benefits in an inspection regimen that involves disturbance of breeding mice, provided the cage is inspected daily.     

Complex housing environment for farmed blue foxes (Vulpes lagopus): use of various resources

The present study was designed to measure the use of various, simultaneously available resources in a complex housing environment in juvenile blue foxes. Twelve blue fox sibling (male–female) pairs were housed in two-section experimental cages from the age of 8 weeks until the age of 7 months (from June to December). Each experimental cage was furnished with two platforms, a nest box, a sand box and a wooden block. This housing set-up provided the foxes with social contact, and an opportunity for oral manipulation, scratching and nesting, as well as the choice of staying on a solid floor material or on an elevated location. The foxes’ behaviour was recorded at three time points during autumn (September, November and December). The foxes used all available resources. The most utilised resource was the nest box, possibly because it could be utilised in several ways (as a shelter, an elevated location, an object for scratching and for oral manipulation). The foxes also stayed more in the cage section containing the nest box than in the cage section containing a sand box. The foxes rested much on the cage floor, but they also used the interior of the nest box and elevated locations for resting. Social contact often occurred during resting. Thus, the nest box and elevated location, in conjunction with social contact seem to be valuable while resting. While active, the foxes utilised the cage floor and roof of the nest box instead of the platforms. Scratching, digging and an interaction with the wooden block were seldom observed. Activity occurred mainly on the ‘empty’ cage area. In conclusion, all studied resources provided blue foxes with a distinct value, as they all were used in the complex housing environment. The nest box is used most and for most variable behaviours.     

The effects of different rack systems on the breeding performance of DBA/2 mice

Housing systems for laboratory animals have been developed over a long time. Micro-environmental systems such as positive, individually ventilated caging systems and forced-air-ventilated systems are increasingly used by many researchers to reduce cross contamination between cages. There have been many investigations of the impact of these systems on the health of animals, the light intensity, the relative humidity and temperature of cages, the concentration of ammonia and CO(2), and other factors in the cages. The aim of the present study was to compare the effects of different rack systems and to understand the influence of environmental enrichment on the breeding performance of mice. Sixty DBA/2 breeding pairs were used for this experiment. Animals were kept in three rack systems: a ventilated cabinet, a normal open rack and an individually ventilated cage rack (IVC rack) with enriched or non-enriched type II elongated Makrolon cages. Reproduction performance was recorded from 10 to 40 weeks of age. In all three rack systems there was a similar breeding index (pups/dam/week) in non-enriched groups during the long-term breeding period, but the coefficients of variation in the IVC rack were higher for most parameters. This type of enrichment seems to lead to a decrease in the number of pups born, especially in the IVC group. However, there was no significant difference in breeding index (young weaned/female/week).     

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.