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We investigated if there were any negative effects on the behaviour and physiology of rats housed in groups of five in two types of enriched cages and compared them with paired-housed rats housed in traditional cages. Eighty-four male Wistar and Sprague-Dawley rats were housed in an enriched rat cage (ERC), a rebuilt rabbit cage (RRC) or a Makrolon III cage (MC) system from 5–16 weeks of age with access to different enrichments. Recordings of behaviour and cage use (3 × 24 h video recording), weekly weighing, measuring food consumption four days/week and water consumption two days/week, were carried out. The rats’ muscle strength was assessed using the ‘inclined plane’ at the end of the study, and after euthanasia the adrenal glands were removed and weighed. Being in the shelter was the most common behaviour in the ERC and RRC groups. In the MC group, which lacked a shelter, rats performed the highest percentage of lying, grooming, rearing, play fighting and manipulating paper shreds. Rats in the RRC had the highest percentage of standing and manipulating gnawing sticks. Water consumption was higher in MC than in ERC and RRC rats. Rats from the RRC managed to remain at a steeper angle on the ‘inclined plane’ than rats from the MC. There were no significant effects of cage type on weight gain, food consumption or relative weights of adrenal glands. In conclusion, male rats kept in groups of five in larger enriched cages benefited from the enrichments, and no negative effects were found in the larger groups. [ABSTRACT FROM PUBLISHER]

Aggressiveness, expressed by fighting, is a frequent problem in group-housed laboratory male mice and results in increased stress, injury, and death. One way to prevent fighting is by pairing the male mice with ovariectomized female mice to provide a compatible companion. However, the effect of these housing conditions remains unclear. Therefore, we aimed to evaluate behavior and stress levels in two different housing conditions, pair-housed with an ovariectomized female and group-housed with other males. Behavioral tests were performed to assess stress and anxiety-like behavior. Moreover, the corticosterone levels in plasma were measured by ELISA. Based on home cage behavior assessment, pair-housed male mice showed no signs of fighting, not even after isolation and regrouping. Our results also showed that the pair-housed males had a better memory and demonstrated less anxiety-like behavior. Subsequently, the pair-housed male mice had a larger reduction in corticosterone levels compared to group-housed males. Overall, pair-housing reduced anxiety-like behavior and stress levels in male mice compared to standard group-housing.

Aggression among group-housed male mice is a major animal welfare concern often observed at animal facilities. Studies designed to understand the causes of male mice aggression have used different methodological approaches and have been heterogeneous, using different strains, environmental enrichments, housing conditions, group formations and durations. By conducting a systematic literature review based on 198 observed conclusions from 90 articles, we showed that the methodological approach used to study aggression was relevant for the outcome and suggested that home cage observations were better when studying home cage aggression than tests provoking aggression outside the home cage. The study further revealed that aggression is a complex problem; one solution will not be appropriate for all animal facilities and all research projects. Recommendations were provided on promising tools to minimize aggression, based on the results, which included what type of environmental enrichments could be appropriate and which strains of male mice were less likely to be aggressive.

Animals used for scientific purposes are protected by EU legislation. Social animals should be kept in stable groups that enable species-typical social behavior and provide individuals with social comfort. However, when group-housing male mice, aggression within the homecage is a common husbandry and welfare problem. Excessive fighting and injuries due to aggression can cause pain and stress, resulting in individuals being euthanized or housed individually. In addition, stress can alter physiological parameters, risking scientific validity and generating larger sample sizes. Mouse aggression, and the consequences thereof, thus opposes the 3R goals of Refining the methods to minimize potential pain and suffering and Reducing the number of animals used. Animal technicians, veterinarians, and scientists using animals have valuable information on how these problems are experienced and handled in practice. We assembled these experiences from laboratory animal facilities in Sweden, mapping problems observed and identifying strategies used to prevent mouse aggression. In line with current literature, less aggression was perceived if mice were grouped before sexual maturity, re-grouping avoided and nesting material transferred at cage cleaning. Preventing aggression will minimize pain and suffering and enable housing of stable groups, leading to more reliable scientific outcomes and is thus of high 3Rs relevance., Competing Interests: The authors declare no conflict of interest.