L.F.M. van Zutphen

Preferences for nesting material as environmental enrichment for laboratory mice

Behavioural and psychological needs of laboratory animals generally cannot adequately be met in standard laboratory cages. Environmental enrichment, which provides a more structured environment can enhance the well-being of laboratory animals. They may perform more of their species-specific behaviour and may control their environment in a better way. An easily applicable form of enrichment for laboratory mice is nesting material. Six different types of nesting materials were evaluated in a preference test with male and female animals of two strains (C57BL/6J or BALB/c, n=48). No significant differences in preference were found between the strains or between the sexes. All mice showed a clear preference for cages with tissues or towels as compared to paper strips or no nesting material, and for cages with cotton string or wood-wool as compared to wood shavings or no nesting material. Paper-derived materials were preferred over wood-derived materials, although the results also suggest that the nature (paper or wood) of the nesting material is less important than its structure, which determines the nestability of the material. Nesting material may be a relatively simple method to contribute to the well-being of laboratory mice.

Male management: coping with aggression problems in male laboratory mice

In a laboratory environment, aggressive interactions between male mice may exceed normal levels leading to negative effects both on the well-being of the animals and on the validity of experimental results. In this paper we review results from the literature and our own research with regard to coping with excessive aggressive behaviour in male laboratory mice. Based on this review practical recommendations concerning the housing and care of male laboratory mice are formulated. In short, it is recommended to avoid individual housing, to transfer odour cues from the nesting area during cage cleaning and to apply nesting material as environmental enrichment. Furthermore, group size should be optimized to three animals per cage. Further research, in particular into the effects of frequency, duration, type and severity of disturbances during an experiment on the degree of aggression, is recommended.

Nesting Material as Environmental Enrichment Has No Adverse Effects on Behavior and Physiology of Laboratory Mice

Environmental enrichment may improve the quality of life of captive animals by altering the environment of animals so that they are able to perform more of the behavior that is within the range of the animals species-specific repertoire. When enrichment is introduced into an animals environment, it is important to evaluate the effect of the enrichment program and to assess whether the animal continues to use the enrichment in the long-term. Groups of mice were housed under either standard or enriched conditions for several weeks. Nesting material which was highly preferred in previous studies was used as enrichment. During the period of differential housing several behavioral parameters (behavioral tests and handling) and physiological parameters (urine and plasma corticosterone, food and water intake, body and adrenal weight) were monitored to determine the impact of environmental enrichment. Observations were made to determine whether or not the mice continued to use the enrichment. The results indicated that throughout the study all mice used the nesting material to build nests and that mice from enriched conditions weighed more than mice housed under standard conditions, although the latter consumed more food. No major differences for behavioral and physiological parameters were found between the groups of mice housed under different conditions. Therefore it is not likely that supply of nesting material will jeopardize the outcome of experiments.

Strength of preference for nesting material as environmental enrichment for laboratory mice

Abstract The present paper describes two experiments in which preferences of laboratory mice for materials which could serve as cage enrichment were investigated. In the first experiment, presence of nesting material (paper towel or tissue) and the presence of a nest box (perforated metal or clear perspex box) were tested against each other. On average, the 47 mice tested spent significantly more time in the cage with the nesting material (more than 69% of their total time, whereas less than 25% of their time in the cage with the nest box). In the second experiment, the preferred nesting material (tissues) was placed in a cage with a grid floor (previously found to be avoided) and the next box (perforated metal) was placed in another cage, connected to the first, with a solid floor covered with sawdust bedding material. In this experiment, 24 female mice were tested and on average they spent more than 67% of their time in the cage with the nesting material, despite the presence of a grid floor. Thus, it is concluded that providing a cage with nesting material (in addition to bedding) may be essential for the well-being of laboratory mice.

Validation of a new system for the automatic registration of behaviour in mice and rats

A newly developed behaviour registration system, Laboratory Animal Behaviour Observation, Registration and Analysis System (LABORAS) for the automatic registration of different behavioural elements of mice and rats was validated. The LABORAS sensor platform records vibrations evoked by animal movements and the LABORAS software translates these into the corresponding behaviours. Data obtained by using LABORAS were compared with data from conventional observation methods (observations of videotapes by human observers). The results indicate that LABORAS is a reliable system for the automated registration of eating, drinking, grooming, climbing, resting and locomotion of mice during a prolonged period of time. In rats, grooming, locomotion and resting also met the pre-defined validation criteria. The system can reduce observation labour and time considerably.

Complete homology maps of the rabbit (Oryctolagus cuniculus) and human by reciprocal chromosome painting

Fluorescence in situ hybridization (FISH) was used to construct a homology map to analyse the extent of evolutionary conservation of chromosome segments between human and rabbit (Oryctolagus cuniculus, 2n = 44). Chromosome-specific probes were established by bivariate fluorescence activated flow sorting followed by degenerate oligonucleotide-primed PCR (DOP-PCR). Painting of rabbit probes to human chromosomes and vice versa allowed a detailed analysis of the homology between these species. All rabbit chromosome paints, except for the Y paint, hybridized to human chromosomes. All human chromosome paints, except for the Y paint, hybridized to rabbit chromosomes. The results obtained revealed extensive genome conservation between the two species. Rabbit chromosomes 12, 19 and X were found to be completely homologous to human chromosomes 6, 17 and X, respectively. All other human chromosomes were homologous to two or sometimes three rabbit chromosomes. Many conserved chromosome segments found previously in other mammals (e.g. cat, pig, cattle, Indian muntjac) were also found to be conserved in rabbit chromosomes.

Preference for social contact versus environmental enrichment in male laboratory mice

Due to their aggressive nature, male mice are less frequently used than female mice in biomedical research. When aggressive males are being used, individual housing is common practice. The question arises whether this is an acceptable housing for a social species. The present study was designed to gain more insight into the nature of inter-male social contact and into the potential of a form of environmental enrichment (nesting material) to compensate for the lack of social contact. In a series of tests, we analysed whether male mice of different ages preferred to spend time (1) near a familiar cage mate versus an empty cage, or (2) near to a familiar cage mate versus direct contact with nesting material (tissues). Dwelling time in each of the test cages and sleeping sites was recorded, as was the behaviour of the test mice. Results indicated that when other conditions were similar, male mice preferred to sleep in close proximity to their familiar cage mate. Furthermore, the need to engage in active social behaviour increased with age. Tissues were used to a large extent for sleeping and sleep-related behaviour. It is concluded that single housing in order to avoid aggression between male mice is a solution with evident negative consequences for the animals. When individual housing is inevitable due to excessive aggressive behaviour, the presence of nesting material could partly compensate for the deprivation of social contact.

Effect of restraint and injection methods on heart rate and body temperature in mice

Routine procedures in the laboratory, inducing acute stress, will have an impact on the animals and might thereby influence scientific results. In an attempt to gain more insight into quantifying this acute stress by means of the parameters heart rate (HR) and body temperature (BT), we subjected mice to different restraint and injection methods. We first compared the treatment response of HR and BT, measured by means of radiotelemetry, with the treatment response of plasma corticosterone (pCORT), a common and well-validated parameter for measuring acute stress responses. It was found that HR, and to a lesser extent also BT, parallels pCORT values after subjecting the animals to different methods of restraint. Secondly, the acute stress response caused by different injection methods was evaluated. Again, HR was found to be a more sensitive parameter than BT. We found that, in case of sham injections, the acute stress response after an intraperitoneal (i.p.) injection was more pronounced than after intramuscular (i.m.) or subcutaneous (s.c.) injections, but this difference was found to be inconsistent when saline was used as injection fluid. In a third experiment we investigated if the level of experience of the animal technician influenced the stress response after s.c. injections, but no differences were found. Overall, the results have indicated that HR might be considered as a useful parameter for measuring acute stress responses to routine procedures, but the value of BT seems to be of limited value in this respect.

Strain differences in response to dietary cholesterol by JAX rabbits: Correlation with esterase patterns

Six strains of genetically defined JAX rabbits were tested for their serum cholesterol levels (total and free) in response to a 0.5% cholesterol diet. Marked differences in response between the 6 strains were found. IIIVO/J and WH/J are low responding strains, X/J and ACEP/J are intermediate responding strains, and OS/J and AX/J are high responding strains. After 4 weeks of the cholesterol diet the total serum cholesterol level of the high responding AX/J strain was about 5-fold greater than the level of the low responding IIIVO/J strain. The esterified/total (E/T) ratio appeared to be higher in the high responding strains, indicating a synergistic effect in the process of atherosclerosis. The response of the individual rabbits to the cholesterol diet was compared with the patterns of serum and liver esterase zymograms. This comparison indicated a correlation of the dietary cholesterol susceptibility with the presence or absence of the esterase zones in the anodal, fast moving region of the gel.

Sterol balance and cholesterol absorption in inbred strains of rabbits hypo- or hyperresponsive to dietary cholesterol

In 2 inbred strains of rabbits with high or low response of plasma cholesterol to dietary cholesterol, excretion of steroids in the feces and efficiency of cholesterol absorption were determined. Rates of whole-body cholesterol synthesis, measured as fecal excretion of bile acids and neutral steroids minus cholesterol intake, were similar in hypo- and hyperresponders fed a low-cholesterol (8 mumol/100 g) diet. Transfer of the rabbits to a high-cholesterol (182 mumol/100 g) diet caused an increase in fecal bile acid excretion in hypo- but not in hyperresponders. Dietary cholesterol did not affect neutral steroid excretion in either rabbit strain. Hyperresponders tended to accumulate more cholesterol in their body than did hyporesponders. After the rabbits were switched back from the high- to the low-cholesterol diet, rates of whole-body cholesterol synthesis were significantly higher in the hypo- than in the hyperresponders. With the use of the simultaneous oral administration of [3H]cholesterol and beta-[14C]sitosterol, hyperresponders were found to absorb significantly higher percentages of cholesterol than hyporesponders. It is concluded that the differences in stimulation of bile acid excretion after cholesterol feeding and the efficiency of cholesterol absorption are important determinants of the phenomenon of hypo- and hyperresponsiveness in the 2 inbred rabbit strains.