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.     

Assessment of the use of two commercially available environmental enrichments by laboratory mice by preference testing

In the field of biomedical research, the demand for standardization of environmental enrichment for laboratory animals is growing. For laboratory mice, a wide variety of environmental enrichment items are commercially available. Most of these comply with the demands for standardization, hygiene and ergonomics. Whether these items also comply with their actual purpose, to enhance the well-being of the mice, is often not assessed scientifically. In this study, we tested the preference of mice for two commercially available nest boxes differing in shape and material: the Shepherd Shack/DesRes (SS/DR) and the Tecniplast Mouse House (TMH), in a simple preference test. To indicate strength of preference, both nest boxes were also tested against a highly preferred nesting material. Preference for the most preferred nest box was investigated further. Our results indicated a strong preference by mice for the SS/DR, but not for the TMH. Furthermore, nesting material was almost always combined with the SS/DR, but not with the TMH. More elaborate testing of the SS/DR in an automated preference test system confirmed that mice spent significantly more time in a cage in which an SS/DR is provided. Differences between both nest boxes are discussed with regard to their attractiveness to mice. It is also argued that enrichment should primarily be developed in concordance with the animals' needs prior to the marketing of enrichment tools.     

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).     

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.     

Impact of environmental enrichment in mice. 1: effect of housing conditions on body weight,organ weights and haematology in different strains

Currently, environmental enrichment is a very common means of improving animal well-being, especially for laboratory animals. Although environmental enrichment seems to be a possible way for improving the well-being of animals, the consideration of housing laboratory animals should not only focus solely on animal well-being, manpower and economics but also on the precision and accuracy of the experimental results. The purpose of the present study was to evaluate the effects of enriched cages (nest box, nesting material, climbing bar) on body weight, haematological data and final organ weights. BALB/c, C57BL/6 and A/J mice, originated from Harlan Winkelmann, were used for the experiments - 16 animals of each strain. Animals at 3 weeks of age were marked and separated randomly to enriched or non-enriched cages, in groups of four, half for each housing condition. Both cages were type III Makrolon cages, only the enriched cages contained a nest box, a wood bar for climbing and nesting material. Animals were kept in a clean animal room under specific pathogen free (SPF) conditions. Body weights were recorded every week. Blood samples were collected at 14 weeks of age (white blood cells (WBC), red blood cells (RBC), haemoglobin (HGB), and haematocrit (HCT) were analysed). At 15 weeks of age, the animals were euthanized by CO(2) in their home cages, and final body weight and organ weights (heart, liver, kidney, adrenal, spleen and uterus) were recorded immediately. Although nearly all the test variables were not affected by environmental enrichment in their mean values, the enriched group showed higher coefficients of variation in many variables, and strain differences of both housing conditions were not consistent. The influences of enrichment were shown to be strain- and test-dependent. Such effects may lead to an increase in the number of animals which is necessary or may change the experimental results, especially when a study, using enriched housing conditions, focuses on strain differences. Since the same enrichment design can result in different influences, a positive or a negative or no adverse effect, due to the strain and the variables studied, researchers need to collect more information before enrichment designs are introduced into experimental plans.     

Cage-change interval preference in mice

Before animal research facilities began using individually ventilated cage (IVC) systems for mice, cages were often changed one or more times per week. When using IVC systems, however, it is standard practice to change cages only once every 2-3 weeks. When deciding how often to change cages, personnel may consider the cost of labor needed to change the cage, as well as the cage type and bedding type, rather than animal preference or concern for animal well-being. The authors carried out a simple preference test in groups of mice. Mice were allowed to choose between an unsoiled cage and cages that had not been changed for 1 d, 7 d or 14 d. When evaluating where mice positioned their nests and the amount of time mice spent in the various cages, the authors found that the mice preferred the unsoiled cage. Younger mice (<150 d old) showed a stronger preference for the unsoiled cage than did older mice (>150 d old). Further studies are warranted to evaluate mice's preferences for cages changed at different intervals and to determine whether prolonging the interval between cage changes has any negative effects on mice.     

Effects of separation of resources on behaviour of high-, medium- and low-ranked hens in furnished cages

In our previous studies, we demonstrated that dominant hens had priority in using the dust bath, resulted in increased competition for the resource. It seemed that the problem was that the resource was placed on one side of the cage (‘localised’). Therefore, we designed a medium-sized furnished cage with a dust bath and nest box on both sides of the cage (‘separated’, MFS). To evaluate the effects of separation of these resources, we compared the behaviour of high-, medium- and low-ranked hens in MFS cage with that in small (SF) and medium furnished (MFL) cages with a localised resource. In total, 150 White Leghorn layers were used. At the age of 17 weeks, the hens were randomly divided into three groups and moved to small furnished cages (SF, 90 cm wide; five birds per cage) and two types of medium furnished cages (180 cm wide; 10 birds per cage) with a nest box and dust bath on both sides (MFS) and a nest box and dust bath on one side of the cage (MFL). The total dust bath and nest box areas per hen were same for the three cages. The dominance hierarchy was determined by observing the aggressive interactions and by this high-, medium- and low-ranked hens in each cage were identified. The behaviour, use of facilities and physical condition of these hens were measured. Data were analysed by using repeated measure ANOVA. A significant interaction between social order and cage design was found in the proportions of time spent in the dust bath and on performing dust-bathing (both P < 0.001), and these proportions tended to be higher in higher-ranked hens in SF and MFL. Conversely, the MFS low-ranked hens tended to use the dust bath more than the SF and MFL low-ranked hens. Thus, hens from each rank used the dust bath equally in MFS, though the MFS high-ranked hens tended to use the resource less than the SF and MFL high-ranked hens. While the frequency of pre-laying sitting was lower among low-ranked hens (P < 0.05), the proportion of time in the nest box was higher among low- than high-ranked hens (P < 0.01). The low-ranked hens spent more time performing escaping, moving and standing in the nest box. In conclusion, it is suggested that separation of the dust bath to two locations would be an effective arrangement to promote more equal usage of the dust bath by hens from each rank in the furnished cages. It was also confirmed in the present study that nest boxes were not only used for laying eggs but also as a refuge by lower ranked hens.     

The effects of different types of individually ventilated caging systems on growing male mice

Ventilation rate and turnover rate of dry air vary among different types of ventilation systems used with individually ventilated cages (IVCs) and can affect the well-being of rodents housed in these cages. The authors compared the effects of two types of IVC systems, forced-air IVCs and motor-free IVCs, on 4-week-old C57Bl/6J male mice. The mice were acclimatized to the cages for 8 d and then monitored for 87 d. Their body weights, food and water consumption and preferred resting areas were recorded. Mice that were housed in motor-free IVCs had a significantly greater increase in body weight than those housed in forced-air IVCs, despite having similar food consumption. Mice in forced-air IVCs had greater water consumption than mice in motor-free IVCs. In addition, mice in forced-air IVCs were more frequently located in the front halves of their cages, whereas mice in motor-free IVCs were located with similar frequency in the front and back halves of their cages, perhaps because of the higher ventilation rate or the location of the air inlets and outlets in the rear of the cage. These results suggest that body weight, food and water consumption and intracage location of growing male mice are influenced by the type of ventilation system used in the cages in which the mice are housed.     

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.     

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.     

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.     

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.     

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.     

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.     

千岛湖栖息地片段化对大山雀营巢资源利用的影响

为了解栖息地片段化对次级洞巢鸟营巢资源利用的影响, 我们于2008年2-8月, 在千岛湖地区选取21个岛屿, 采用悬挂人工巢箱方法进行大山雀(Parus major)招引实验。通过测定岛屿面积、岛屿隔离度、捕食者活动率、人工巢箱周围植被盖度和人工巢箱巢向等5种参数, 分析大山雀人工巢箱利用与上述参数间的关系。实验期间, 共悬挂443个人工巢箱, 其中有72个(16.3%)被大山雀利用。大山雀倾向于选择捕食者活动率低、植被盖度低、巢口向东或向南的人工巢箱; 岛屿面积和隔离度对大山雀利用人工巢箱不存在显著的直接影响, 但岛屿面积可通过影响捕食者的活动率, 对大山雀利用人工巢箱产生间接影响。因此, 我们认为在栖息地片段化后, 应更多关注对洞巢鸟营巢资源利用有直接影响的栖息地特征因素, 以及片段化效应的间接影响。     

Response of adult New Zealand white rabbits to enrichment objects and paired housing.

Enhancing the psychological well-being of laboratory animals has received much attention recently. Although many studies have been undertaken to determine the effects of cage enrichment techniques on dogs and nonhuman primates, other than scant empirical observations, little has been done to measure these events objectively in lagomorphs. We studied adult female New Zealand White (NZW) rabbits to learn if, when given the opportunity, individual rabbits would use different enrichment objects placed in their cages, and to determine if rabbits preferred to be in proximity to one another, or apart. Three different objects were evaluated with eight rabbits individually housed in conventional cages. Each object introduced into individual rabbit cages stimulated substantial interaction, especially chewing behavior. Eight other rabbits were pair-housed in a modified caging system with a special access port between two separate cages. When given a choice, rabbits preferred to be in the same cage with other rabbits. In both studies, individual behaviors were monitored, as well as either the type of interaction and percentage of observations spent with each object or, in the housing study, percentage of observations involved with different types of activity, and relative location of the paired rabbits.     

Nest box use by woodland dormice (Graphiurus murinus): the influence of life cycle and nest box placement

The use of nest boxes by the woodland dormouse, Graphiurus murinus, was investigated over a 13-month period in a riverine forest of the Great Fish River Reserve, South Africa. We predicted that some characteristics of nest box placement would affect nest box use and that the seasonal pattern of nest box use would be linked to the species' life cycle and physiological and socioecological characteristics. Generalized linear models indicated that the time since nest box installation and nest box height above ground positively affected the frequency and intensity of nest box use. Male and female dormice, as well as adults and juveniles, did not differ in the number of nest boxes used and equally occupied individual nest boxes. The percentage of nest boxes used peaked during spring and summer (breeding period) and dropped during winter (hibernation). However, whereas significantly more males were caught during the mating season (spring), the number of females occupying nest boxes was constant during the year. As female dormice successfully bred in the nest boxes, the observed sexual patterns suggest that (artificial) nest sites represent an important resource for females, whereas females seem to constitute the main resource for males, as predicted by the socioecological model.     

The effect of cage size on reproductive performance and behavior of C57BL/6 mice

Scientific research has yet to conclusively determine the optimal cage size for mice. The authors examined the effect of cage size on mouse breeding performance and on offspring behavior, which can serve as indications of overall well-being. They housed breeding trios of C57BL/6Tac mice in standard or large individually ventilated cages and measured four reproductive parameters: litter size; litter survival to weaning age; average pup weight at 7, 14 and 21 days; and the number of days between litter births. They investigated the behavior of a subset of male and female pups from parents housed in cages of each size in the elevated plus maze test, the open field assay and the acoustic startle test. Cage size had no significant effect on any of the reproductive parameters measured and few or inconsistent effects on behavior in weaned pups.     

Carbon dioxide concentrations in unventilated IVC cages

The use of individually ventilated cage (IVC) systems has become more common worldwide. The various systems are becoming more and more sealed in order to protect the animals against infections and the staff against allergens; which, however, may lead to problematic CO2 concentrations, if the cages are left unventilated. In this study it is shown that, depending on how tight the cage is and the number of animals housed in each cage, CO2 inside the cage within 2 h will increase to levels of between 2 and 8%.