Aspen wood-wool is preferred as a resting place, but does not affect intracage fighting of male BALB/c and C57BL/6J mice

Aspen wood-wool, provided as nesting material, was evaluated as a possible improvement of cage environment for 10-14-week-old inbred male mice maintained in groups of six (BALB/c n = 72 and C57BL/6J n = 36). The daily behaviour of mice was video recorded and their body weight, food consumption, weights of some organs and serum corticosterone concentrations were measured. Aggressive interactions between cage mates and against a strange intruder as well as the number of wounds on the back of the animals was monitored in order to evaluate the effect of nesting material on intermale aggression. Nesting material did not affect the daily active/passive behaviour patterns of mice, although animals clearly preferred it as a resting place. BALB/c mice given nesting material showed less weight gain and smaller brown adipose tissue weights than animals without nesting material. The other characteristics measured were not affected by the presence of nesting material in either strain. The presence of nesting material had no effect on fighting in cages. C57BL/6J mice were more aggressive than BALB/c mice according to the number of wounded animals in a cage. Wounded BALB/c mice had enlarged spleens and decreased epididymal adipose tissue weights. In conclusion, the nesting material used in this study did not adversely affect the animals. On the other hand, the material was clearly preferred to conventional bedding as a resting place. These findings suggest that nesting material may improve the cage environment of laboratory mice. Furthermore, there was an indication of strain differences in aggressive behaviour. It could be suggested that C57BL/6J mice are less tolerant towards intruders and housing six mice per cage is not suitable for this strain.     

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.     

Aggressive behavior in female golden hamsters: development and the effect of repeated social stress

In male golden hamsters, agonistic behavior matures during puberty, changing from play fighting to adult-like aggression. In addition, this transition is accelerated by repeated social subjugation early in puberty. However, little is known about the development of agonistic behavior in females. In the present study, we compared the development of agonistic behavior in male and female golden hamsters. Furthermore, we also tested the effects of repeated social subjugation on the development of agonistic behavior during puberty. Hamsters were tested for agonistic behavior in the presence of a smaller intruder at different intervals during puberty. Several observations were made. First, the frequency of attacks remained stable in females, while varying in males. Second, the transition from play fighting to adult-like aggression occurred at earlier time periods in females than in males. Finally, a clear transitional period marked by attacks focused on the flanks was observable in males around mid-puberty. However, this transitional period was not apparent in females. In addition, juvenile females were exposed to aggressive adult males or females. In both cases, repeated exposure to stress had no statistically significant effect on the development of agonistic behavior. After 2 weeks of subjugation, exposure to aggressive adults had no effect on serum cortisol levels, indicating that juvenile females habituate to repeated social stress. These data show significant sex differences in the development of agonistic behavior and adaptation to repeated stress in juvenile golden hamsters.     

Fighting in NIH/S male mice: consequences for behaviour in resident-intruder tests and physiological parameters

Fighting is known to occur frequently in male mouse groups. In this study with outbred NIH/S mice, the possible impact of individual aggressiveness on fighting in groups and on the social status of animals was studied. Male mice were pre-tested in a resident-intruder (RI) test and rated as initially aggressive or non-aggressive according to their attack behaviour against an intruder. Thereafter they were randomly allocated to new social groups, with four mice per cage. Fighting in groups was increased when several initially aggressive animals were included in the group. Within the groups, animals were rated as dominants and subordinates according to their behaviour toward a strange intruder introduced into their home-cage (Group Intruder, GI) test and the occurrence of wounds. Additionally, subordinates were divided into aggressive and non-aggressive categories according to their behaviour in the second RI test, which was performed 3 weeks after grouping. The behaviour in the RI test prior to group-housing did not predict the individual social status or possibility of being wounded in the new social environment. On the other hand, the social relationships in the new group affected the behaviour in a subsequent RI test. All dominants showed aggressive behaviour during the second RI test. Those subordinates which behaved aggressively during this test received the most numerous and serious wounds, suggesting that in the new groups their interactions with the other group members were mostly aggressive. The reduced weight of epididymal adipose tissue in dominant and aggressive subordinates may indicate that they had fought continuously. Social status or levels of fighting in a group did not affect individual weight gain or the other physiological parameters measured. The wounded animals had enlarged spleens and reduced weights of epididymal adipose tissue, which were probably the results of increased activity of the immune system and reduced welfare, respectively. In conclusion, individual aggressiveness seems to be greatly affected by the demands of the social environment. Fighting in mouse groups leading to wounded animals may have effects on physiological research parameters.     

Female Intrasexual Territoriality and its Potential Adaptive Significance: The Pampean Grassland Mouse as an Ecological Model Species

Territorial behaviour in female small mammals has been proposed as a mechanism to defend limited ecological resources or their pups against conspecific infanticidal or predators. Female territorial behaviour very often is associated with reproductive activity due to the fact that frequency and intensity of aggression are exhibited mainly when females are pregnant or lactating. In vole and mice species, female territoriality would be a counterstrategy to prevent the killing of their pups by conspecific breeding females. To study whether female territoriality is a strategy for pups or nest defence against infanticidal breeding females, and whether time invested in nursing young affects aggressive response of mothers, we used the Pampean grassland mouse (Akodon azarae) as an ecological model species. We conducted resident–intruder tests between lactating females. Differences in residency time (48 vs. 72 h) of focal females in their home territory were also included in the analysis. In all cases, the pups of both resident and intruder mothers were placed with the nesting material from their reproductive cages. Resident mothers were always more aggressive than intruders and they were even more aggressive when they spent more time nursing their pups. Contrarily, intruder females exhibited the greatest values of submissive behaviours. Our results show that female territoriality of A. azarae would represent a strategy to protect pups from potentially infanticidal females. We discuss the extent of female intrasexual territoriality and its potential adaptive significance in relation to strategies which lead to increase their reproductive success.     

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.     

Two Sides of the Same Coin? Consistency in Aggression to Conspecifics and Predators in a Female Songbird

Different forms of aggression have traditionally been treated separately according to function or context (e.g. aggression towards a conspecific versus a predator). However, recent work on individual consistency in behavior predicts that different forms of aggression may be correlated across contexts, suggesting a lack of independence. For nesting birds, aggression towards both conspecifics and nest predators can affect reproductive success, yet the relationship between these behaviors, especially in females, is not known. Here we examine free-living female dark-eyed juncos (Junco hyemalis) and compare their aggressive responses towards three types of simulated intruders near the nest: a same-sex conspecific, an opposite-sex conspecific, and a nest predator. We also examine differences in the strength of response that might relate to the immediacy of the perceived threat the intruder poses for the female or her offspring. We found greater aggression directed towards a predator than a same-sex intruder, and towards a same-sex than an opposite-sex intruder, consistent with a predator being a more immediate threat than a same-sex intruder, followed by an opposite-sex intruder. We also found positive relationships across individuals between responses to a same-sex intruder and a simulated predator, and between responses to a same-sex and an opposite-sex intruder, indicating that individual females are consistent in their relative level of aggression across contexts. If correlated behaviors are mediated by related mechanisms, then different forms of aggression may be expressions of the same behavioral tendency and constrained from evolving independently.     

Sickness and Aggressive Behavior in Dominant and Subordinate Mice*

Sick animals show a set of organized behavioral changes (sickness behavior), which is the result of a motivational re‐organization of the behavior as a whole. Sickness behavior display can be influenced by the social context. In this work, we sought to investigate the regulation of sickness behavior within a pair of mice in the presence of an intruder mouse. Dominant and subordinate mice were treated with the bacterial endotoxin lipopolysaccharide (LPS) and were challenged with the presence of an intruder mouse. LPS effects depended on ranking and social context. Even though dominant mice displayed more agonistic interaction towards the intruder, subordinate mice displayed agonistic behavior towards the intruder when their dominant companion was treated with LPS. The results show that, not only sickness behavior is differentially expressed among different social ranks, but also that sickness behavior is related to different reactions among surrounding animals. These data are relevant for a biological approach to the relation between sickness behavior and social behavior.     

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.     

Social Experience During Development and Female Offspring Aggression in Peromyscus Mice

Cross‐fostering between the highly aggressive, biparental California mouse (Peromyscus californicus) and the less aggressive, less parental white‐footed mouse (P. leucopus) influences female offspring attack latency in California mice, but not in white‐footed mice. Adult female California mice raised by white‐footed mice expressed longer attack latencies in a neutral‐arena test but not in a resident‐intruder test. One social cue that may be used by offspring to develop environmentally appropriate levels of aggression is the type of parental care during development. In California mice, a composite score of maternal behavior was positively associated with neutral‐arena aggression as indicated by decreased attack latency. In both species, paternal nest‐building was positively associated with neutral‐arena aggression and higher maternal retrieval behavior predicted higher offspring resident‐intruder aggression as indicated by decreased attack latency. Together, these results indicate that parental behavior has the potential to shape the development of attack latency in female offspring.     

Lateralization of Aggression during Reproduction in Male Siamese Fighting Fish

A preference for the left‐eye use during aggressive interactions has been widely reported in the literature, even though in some cases the direction of lateralization varies among individuals within populations. Laterality of aggression in male Siamese fighting fish has been described in a number of studies, yet very little is known about lateralization of aggression during reproduction and/or parental care in fish. Here, we aimed to investigate the relationship between the different reproductive phases and lateralization in eye use during aggressive interactions in males of Siamese fighting fish. Lateralization in eye use is influenced during the early reproductive state, before and after the bubble nest construction stages. We found that nest‐holding males preferentially used the right eye before and after bubble nest construction independent of the sex of the intruder. During the later reproductive phases, aggressiveness increased whereby the direction of lateralization rather than the degree was influenced supporting the hypothesis that reproductive state influences behavioral consistency in Siamese fighting fish.     

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.     

Circadian wheel-running activity rhythms are unaltered by acute cage enrichment with nests and shelters

C57BL/6J inbred male mice were assessed for the effects of cotton nesting material on circadian running-wheel assays at 60, 157 and 222 days of age. The oldest mice were also tested for effects of access to plastic igloo shelters and the combination of cotton plus igloos. All mice were assayed for mean activity level, acrophase and entrained circadian period in 12:12 LD, and mean activity level and free-running period in DD for circadian wheel-running activity. There were no significant effects, at any age, of either nesting material or shelters, or the combination of both, on any measures of circadian running-wheel activity. We conclude that circadian rhythm assessment is unlikely to be significantly altered by the inclusion of enrichment materials in running wheel cages.     

Neurobiological correlates premeditated (learned) aggression: seeking new experimental approaches]

Theoretical possibility of experimental modeling of learned (premediated) aggression developing in human after experience of aggression is considered. The sensory contact technique increases aggressiveness in male mice and allows aggressive type of behavior to be formed as a result of repeated experience of victories in daily agonistic confrontations. Some behavioral domains confirm the development of learned aggression in males similar to those in humans. The features are: repeated experience of aggression reinforced by victories; elements of learned behavior after period of confrontations; intent, measured by increase of the aggressive motivation prior agonistic confrontation; decreased emotionality estimated by parameters of open field behavior. Relevant stimuli provoke demonstration of aggression. This review summarized data on the influence of positive fighting experience in daily intermale confrontations on the behavior, neurochemistry and physiology of aggressive mice (winners). This sort of experience changes many characteristics in individual and social behaviors, these having been estimated in different tests and in varied situations. Some physiological parameters are also changed in the winners. Neurochemical data confirm the activation of brain dopaminergic systems and functional inhibition of serotonergic system in winners under influence of repeated experience of aggression. The expression of the neurochemical and behavioral changes observed in winners has been found dependent on the mouse strain and on the duration of their agonistic confrontations. Similarities in mechanisms of learned aggression in humans and mice are considered.     

Visual Assessment of Relative Body Size in a Cichlid Fish,the Oscar,Astronotus ocellatus

In cichlid fishes, fighting ability, success at nest defense, and fecundity are all positively correlated with body size. Selection should therefore favor the evolution of strategies for the assessment of relative body size during intraspecific interactions. The effect of relative body size on agonistic behavior in the oscar (Astronotus ocellatus) was tested using dummies. As predicted by game theory models, attack was strongly correlated with the size of the dummy relative to the subject. The most intense aggression occurred when dummy size was approximately 75% of subject size. These data support the prediciton that perceived body size asymmetries are important determinants of agonistic behavior in these fishes. Further, the results suggest that oscars can visually assess relative body size, and may estimate relative fighting ability without tests of strength. Subject size was positively correlated with intensity of aggressive behavior. Attack and display frequencies were positively correlated with a subject's body size, while latencies to approach and attack dummies were negatively correlated with subject size.     

Social and Thermal Cues Influence Nest‐site Selection in a Nocturnal Gecko,Oedura lesueurii

In oviparous species lacking parental care, successful reproduction depends on females selecting nest sites that facilitate embryonic development. Such sites may be limited in the environment, which can lead to multiple females using the same nest site simultaneously. However, there are several alternative explanations for communal nesting, including natal homing, predator satiation, and adaptive benefits to offspring. We used laboratory experiments to evaluate three hypotheses about nest‐site selection in velvet geckos (Oedura lesueurii), which often nest communally. We investigated whether the trend to nest communally is influenced by the following: (1) evidence of previous nesting (hatched eggshells); (2) body size; and/or (3) thermal regimes. When given the choice, females laid their eggs in shelters containing hatched eggshells rather than in empty shelters, and this was not influenced by body size. Females selected nest sites that were cooler than their own mean selected body temperatures, suggesting that thermal requirements of their developing embryos could outweigh their own thermoregulatory preferences. Field observations of natal homing and high predation rates on gravid females suggest that imprinting on nest sites and/or predator swamping also play roles in communal nesting. Collectively, our results suggest that female velvet geckos use multiple cues to select appropriate nest sites, and hence that multiple mechanisms result in communal nesting behavior in this species.     

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.     

Behaviour of laboratory mice in different housing conditions when allowed to self-administer an anxiolytic

Standard cages prevent mice from performing several natural behaviours for which they are motivated. As a consequence, abnormal behaviours sometimes develop and mice often spend long periods inactive. To improve welfare, cages are sometimes furnished with items such as nesting material, shelters and running wheels. We have previously reported that when allowed to self-administer an anxiolytic, mice in furnished cages consume less anxiolytic than mice in standard cages. This paper presents the results of behaviour studies of the mice in the same experiment. Female C57BL/6J mice (3 per cage) were housed in Standard (n = 10), Unpredictable (n = 10) or Furnished (n = 6) cages. Unpredictable cages were identical to Standard cages, but were exposed to unpredictable events two to three times a week. Furnished cages were double the size of Standard cages and contained nesting material, nest box, tubes, chew blocks and a running wheel. During three consecutive periods, mice had access to only water (control), water or an anxiolytic solution on a daily alternating schedule (forced consumption), and finally, both water and anxiolytic (self-administration). Behaviour was analysed from video recordings taken during the dark phase. The housing type affected behaviour both under the control and the self-administration conditions. Overall, mice in Furnished cages spent less time resting and performing bar-related behaviours and more time on exploratory/locomotory behaviours. Mice in Furnished cages also performed less bar-circling stereotypies than mice in Standard cages. The Unpredictable treatment did not significantly affect behaviour compared to mice in the Standard conditions. There was an overall effect of anxiolytic availability on rest-related behaviours and on exploration-locomotion behaviours, in that mice rested more and spent less time on exploration and locomotion when they were able to self-administer the anxiolytic.     

Assessing nest building in mice

For small rodents, nests are important in heat conservation as well as reproduction and shelter. Nesting is easily measured in the home cages of mice, particularly with the advent of pressed cotton materials. The mice first shred the tightly packed material, then arrange it into a nest. Published studies have often used materials such as hay, twine or tissues, sometimes preshredded, and have assigned scores of the quality of the resulting nest with rather rudimentary rating scales; e.g., 0, no nest; 1, flat nest; 2, nest covering the mouse. The protocol described here uses pressed cotton squares and a definitive 5-point nest-rating scale. Any unshredded material left after a bout of nesting can also be weighed, providing a semi-independent objective assay of nesting ability. Nesting has been shown to be sensitive to brain lesions, pharmacological agents and genetic mutations. This is a simple, cheap and easily done test that, along with other tests of species-typical behavior, is a sensitive assay for identifying previously unknown behavioral phenotypes. The test needs to be done overnight, but it should take no more than 5 minutes to set up plus 1 minute to assess one nest and weigh the untorn residue.     

Sex-differences in nest defence in Fieldfares Turdus pilaris in relation to their size and physical condition

The nest defence of solitary-breeding Fieldfares Turdus pilaris against a human intruder, a corvid and a hawk was studied in a mountain birch forest in central Norway in 1990–2002. Females defended their nests against a human intruder more vigorously than did males before hatching, and vice versa after hatching. Although both sexes increased their nest defence, only males showed a significant increase and showed a greater defence than females at the chick stage. Females with 1-week-old nestlings had longer wings, were heavier and in a better physical condition than those that failed to get nestlings to that stage. Females, but not males, lost condition during nesting. Birds in better condition were more aggressive. The pattern of nest defence by males and females against the corvid was similar to that shown against human intruders, whereas males took greater risks than females towards the raptor. There was weak evidence that birds that were more successful showed greater levels of nest defence. The sex-role reversal in nest defence may be connected to differences in costs in relation to the renesting potential and to the physical condition of the sexes.