Association between exploratory activity and social individuality in genetically identical mice living in the same enriched environment.

We previously reported that inbred, genetically identical mice living in one enriched environment develop individual behavioral trajectories, indicating increasingly different levels of spatial exploratory behavior as quantified by roaming entropy. Cumulative roaming entropy (cRE) correlated positively with adult hippocampal neurogenesis, a type of plasticity involved in the flexible integration of new information into existing contexts (Freund et al., 2013). The study on which we report here was done in parallel to that first experiment, but here we acquired detailed observational data on the behavior of individual mice. Roaming entropy (RE) was again assessed in real-time with an antenna-based system over the entire experimental period of 3months. Compared to the least active mice in the enclosure (low number of antenna contacts), the most active animals showed tendencies of increased socially interactive behavior in the final observation block whereas least active mice displayed more self-related behavior (non-social local exploration and play). When looking at roaming behavior, we discovered that RE correlated negatively with latent factors representing social exploratory and non-social exploratory and play behavior. Adult neurogenesis could not be studied in the present cohort but we do know that under identical conditions, cumulative RE correlated positively with adult hippocampal neurogenesis. We can thus hypothesize that the mice with more exploratory experience in terms of areal coverage (as quantified by RE) and related greater levels of adult hippocampal plasticity, might also be the ones that were less involved in interactions within the group and, hence, more individualistic. While this remains to be confirmed experimentally, the present data suggest that the described mechanism of individualization, which has previously been shown to be hippocampus-dependent, has a social component.

Serotonin transporter knockout and repeated social defeat stress: impact on neuronal morphology and plasticity in limbic brain areas.

Low expression of the human serotonin transporter (5-HTT) gene presumably interacts with stressful life events enhancing susceptibility for affective disorders. 5-Htt knockout (KO) mice display an anxious phenotype, and behavioural differences compared to wild-type (WT) mice are exacerbated after repeated loser experience in a resident-intruder stress paradigm. To assess whether genotype-dependent and stress-induced behavioural differences are reflected in alterations of neuronal morphology in limbic areas, we studied dendritic length and complexity of pyramidal neurons in the anterior cingulate and infralimbic cortices (CG, IL), hippocampus CA1 region, and of pyramidal neurons and interneurons in the lateral (La) and basolateral (BL) amygdaloid nuclei in Golgi-Cox-stained brains of male WT and 5-Htt KO control and loser mice. Spine density was analysed for IL apical and amygdaloid apical and basal pyramidal neuron dendrites. While group differences were absent for parameters analysed in CG, CA1 and amygdaloid interneurons, pyramidal neurons in the IL displayed tendencies to shorter and less spinous distal apical dendrites in 5-Htt KO controls, and to extended proximal dendrites in WT losers compared to WT controls. In contrast, spine density of several dendritic compartments of amygdaloid pyramids was significantly higher in 5-Htt KO mice compared to WT controls. While a tendency to increased spine density was observed in the same dendritic compartments in WT after stress, changes were lacking in stressed compared to control 5-Htt KO mice. Our findings indicate that disturbed 5-HT homeostasis results in alterations of limbic neuronal morphology, especially in higher spinogenesis in amygdaloid pyramidal neurons. Social stress leads to similar but less pronounced changes in the WT, and neuroplasticity upon stress is reduced in 5-Htt KO mice.

Altered phosphorylation but no neurodegeneration in a mouse model of tau hyperphosphorylation.

The role of hyperphosphorylation of tau in Alzheimer's disease is still unsolved. Here we describe a novel transgenic mouse model, expressing a pseudohyperphosphorylated (PHP) variant of the longest human CNS tau isoform in forebrain neurons. We report that pseudohyperphosphorylation decreases phosphorylation at T205 while other sites (T212, S262) are less or not affected compared to mice expressing wildtype tau. Despite the differences in phosphorylation, the subcellular distribution of tau is not affected and mice do not develop highly aggregated states of tau. PHP tau expressing mice do not show any evidence for neurodegeneration as determined from morphometric measurements of neocortical regions, caspase activation, analysis of mitochondrial dysfunction, or determination of spine densities. In agreement, no differences in learning and memory are observed. The data indicates that moderate levels of modified tau alone are not sufficient to induce tau aggregation or neurodegeneration in transgenic mice. With our model it becomes possible to study the effects of hyperphosphorylation at conditions which may prevail in an early preaggregation state of the disease.

Environmental bias? Effects of housing conditions, laboratory environment and experimenter on behavioral tests.

Behavioral testing does not always yield similar results when replicated in different laboratories, and it usually remains unclear whether the variability in results is caused by different laboratory environments or different experimenters conducting the tests. In our study, we applied a systematic variation of housing conditions, laboratories and experimenters in order to test the influence of these variables on the outcome of behavioral tests. We wanted to know whether known effects of different housing conditions on behavior can be demonstrated regardless of the respective laboratory and experimenters. In this study, we compared the behavior of mice kept under enriched housing conditions with mice kept in unstructured cages regarding their exploratory, locomotor and anxiety-related behavior in the barrier test, in the open-field test and in the elevated plus-maze test. Experiments were conducted by six different persons in two different laboratories. In spite of an extensive protocol standardizing laboratory environment, animal maintenance and testing procedures, significant differences in absolute values between different laboratories as well as between different experimenters were noticed in the barrier test and in the elevated plus-maze test but not in the open-field test. However, with regard to the differences between enriched and unstructured housing conditions, overall consistent results were achieved by different experimenters in both laboratories. We conclude that the reliability of behavioral phenotyping is not challenged seriously by experimenter and laboratory environment as long as appropriate standardizations are met and suitable controls are involved.

Role of a neuronal small non-messenger RNA: behavioural alterations in BC1 RNA-deleted mice.

BC1 RNA is a small non-messenger RNA common in dendritic microdomains of neurons in rodents. In order to investigate its possible role in learning and behaviour, we compared controls and knockout mice from three independent founder lines established from separate embryonic stem cells. Mutant mice were healthy with normal brain morphology and appeared to have no neurological deficits. A series of tests for exploration and spatial memory was carried out in three different laboratories. The tests were chosen as to ensure that different aspects of spatial memory and exploration could be separated and that possible effects of confounding variables could be minimised. Exploration was studied in a barrier test, in an open-field test, and in an elevated plus-maze test. Spatial memory was investigated in a Barnes maze and in a Morris water maze (memory for a single location), in a multiple T-maze and in a complex alley maze (route learning), and in a radial maze (working memory). In addition to these laboratory tasks, exploratory behaviour and spatial memory were assessed under semi-naturalistic conditions in a large outdoor pen. The combined results indicate that BC1 RNA-deficient animals show behavioural changes best interpreted in terms of reduced exploration and increased anxiety. In contrast, spatial memory was not affected. In the outdoor pen, the survival rates of BC1-depleted mice were lower than in controls. Thus, we conclude that the neuron-specific non-messenger BC1 RNA contributes to the aptive modulation of behaviour.

Early social stress in male Guinea-pigs changes social behaviour, and autonomic and neuroendocrine functions.

The present study was undertaken to investigate the effects of pre- and early postnatal social stress on the functioning of hormonal, autonomic and behavioural systems, by studying the distribution of sex hormone receptors in limbic brain systems. Dams had either lived in groups with a constant composition (= stable social environment) or in groups with changing compositions, i.e. every third day, two females from different groups were exchanged (= unstable social environment). The subjects were male offspring of dams who had either lived in a stable social environment during pregnancy and lactation (= control males) or in an unstable social environment during this period of life (= early stressed males). From days 20-80, the spontaneous behaviour of control males and early stressed males was recorded in their home cages. Five control males and five early stressed males were killed at 75 days, and five control males and five early stressed males at 120 days. Blood samples were taken to determine serum concentrations of cortisol, testosterone, dehydroepiandrosterone, dehydroepiandrosterone sulphate and oestrogen. The adrenals were prepared to determine tyrosine hydroxylase activities and the brains were used to investigate the distribution of sex-hormone receptors in specific hypothalamic and hippocampal brain areas. Early stressed males showed a behavioural infantilization that was accompanied by significantly decreased adrenal tyrosine hydroxylase activities and dehydroepiandrosterone levels. Furthermore, early stressed males showed a down-regulation of androgen receptors in the medial preoptic area and the nucleus arcuatus of the hypothalamus, as well as of oestrogen receptor alpha in the hippocampus compared to control males. Thus, the present study provides clear evidence that early social stress induces changes in endocrine, autonomic and limbic brain function, which is mirrored by changes in male social behaviour.

Different types of oestrous cycle in two closely related South American rodents (Cavia aperea and Galea musteloides) with different social and mating systems.

A comparative approach was used to investigate two closely related South American rodent species: the wild cavy (Cavia aperea) and the yellow-toothed cavy (Galea musteloides). These species of wild guinea-pig inhabit different habitats and show divergent social and mating systems. Cavia have a polygynous mating system, whereas in Galea promiscuous mating occurs. These observations correspond to functional variations in sperm characteristics, which might be interpreted as adaptations to different reproductive patterns in the females of both species. Twenty-two female C. aperea and G. musteloides (11 of each species) were investigated to elucidate differences in the characteristics of the oestrous cycles. The experimental design consisted of three stages, in which the degree of contact between the sexes was successively changed. During these stages, the reproductive cycle of the females was monitored by examining the condition of the vaginal closure membrane, analysing vaginal smears and determining serum concentrations of oestrogens and progesterone. The results revealed different types of oestrous cycle in the two species. Female C. aperea showed periodical cycles with spontaneously occurring oestrous periods, ovulation and corpus luteum activity. In contrast, in female G. musteloides, oestrus was exclusively induced by the presence of a male. However, after the induction of oestrus, ovulation and corpus luteum activity occurred spontaneously without stimuli from copulation. To date this type of oestrous cycle has not been described for any other mammalian species. Thus, these data indicate that differences in the reproductive patterns of the females may have shaped the evolution of different sperm characteristics in the males.

Social stress during pregnancy and lactation affects in guinea pigs the male offsprings' endocrine status and infantilizes their behaviour.

This study investigated the influence of the social environment during pregnancy and lactation on the male offsprings' endocrine parameters and behaviour in guinea pigs. The subjects studied were sons whose mothers had either lived in a stable social environment during pregnancy and lactation or in an unstable social environment during this period of life. The stable social environment was made by keeping the group composition (one male, five females) constant; in the unstable social environment situation every third day, two females from different groups were exchanged. After weaning, seven groups of sons, whose mothers had lived in an unstable social environment and seven groups of sons, whose mothers had lived in a stable social environment, consisting of two males each, were established. From their 20th through their 100th day of age the spontaneous behaviour of the males was recorded in their home cages. On the endocrine level, cortisol- and testosterone-concentrations in serum as well as adrenal tyrosine hydroxylase activities were determined. Sons, whose mothers had lived in an unstable social environment, showed significantly higher amounts of resting with bodily contact than sons, whose mothers had lived in a stable social environment. Additionally, they displayed this behaviour to an older age than the latter sons. Further on, sons, whose mothers had lived in an unstable social environment, displayed significantly higher frequencies of courtship behaviour than sons, whose mothers had lived in a stable social environment, which was integrated into play behaviour. These behavioural data point to an infantilization of sons, whose mothers had lived in an unstable social environment. The behavioural patterns corresponded with a delayed development of the hypothalamo-pituitary-adrenocortical-axis and a lower activity of the sympathetic-adrenomedullary-system compared to sons whose mothers had lived in a stable social environment. Testosterone did not differ between both categories of males. Thus, the instability of the social environment during pregnancy and lactation had distinct and not yet described effects on the behaviour and endocrine system of the male offspring.

Effects of ACTH applications during pregnancy on the female offsprings' endocrine status and behavior in guinea pigs.

The present study was conducted to elucidate the effects of ACTH application during pregnancy on the female offsprings' endocrine status and behavior in a precocial animal, the guinea pig. We, therefore, studied daughters whose mothers were treated three times with either ACTH-depot or placebo during the sensitive phase of central nervous sexual differentiation of the foetus. From their 41st through their 80th day of age the spontaneous behavior of the daughters was recorded in their home cages, and concentrations of serum cortisol were determined from the 20th through the 90th day of age every 14 days. Daughters whose mothers had received ACTH displayed significantly lower amounts of a defensive aggressive behavior pattern, the urine spray, and they had higher cortisol concentrations than daughters whose mothers had received placebo. Thus, the number of species that shows consequences of an ACTH treatment during pregnancy for the offsprings' endocrine status and behavior is extended to include a precocial animal, the guinea pig.

Maturation of epididymal spermatozoa in the nondomesticated guinea pigs Cavia aperea and Galea musteloides.

The physiological changes occurring in spermatozoa in the male reproductive tract of 2 nondomesticated species of South American guinea pigs with different mating systems were studied. Cavia aperea, the wild ancestor of the domesticated guinea pig, has a polygynous mating system, whereas Galea musteloides exhibits promiscuous mating behavior. The epididymis of both species resembled that of the domesticated guinea pig, with a swathe of tubule convolutions (linking the 2 major parts of the organ) that was of smaller size in Cavia but not Galea. Higher relative epididymal weight was demonstrated in the promiscuous species. During their journey through the epididymis, spermatozoa from Galea developed their potential for motility expression more proximally than did those of Cavia, but motility developed into forward progression in the same region in both species. The maximal velocities exhibited by mature Cavia sperm in vitro were greater than those of Galea. Spermatozoa from Cavia were twice the length of those from Galea, they had larger heads, and the acrosomes of single sperm were more sensitive to disruption during morphological preparation. Only in Cavia did agglutination of sperm into rouleaux occur, after the potential for motility had been developed. Migration of the cytoplasmic droplet along the midpiece occurred in the same regions in both species and before agglutination in Cavia. It is suggested that the male's reproductive strategy (polygyny vs promiscuity) dictates the size of the testis and epididymis, whereas the female's reproductive physiology (induced ovulation vs cyclicity) influences the posttesticular development of sperm morphology and motility in the epididymis.

[Voluntary testing procedures of farm animal housing equipment according to the Animal Welfare Act of 1998]. / Freiwilliges Prüfverfahren für Stalleinrichtungen entsprechend dem novellierten Tierschutzgesetz von 1998.

Before its broad application in practice, housing equipment should be tested, in particular with regard to animal welfare. The differing positions of the German Federal Council (Bundesrat) and the German Federal Parliament (Bundestag), whether such testing should be mandatory or voluntary, have been conciliated in the amended animal welfare act by empowering the Federal Ministry of Food, Agriculture and Fisheries (BML) to fix official standards for voluntary testing procedures by regulation. On request of the BML, a report as scientific basis for a draft regulation is currently prepared by the scientific animal welfare committee of the German Agricultural Society (DLG). The scientific animal welfare committee has been appointed by the DLG in order to provide support in the effort to strengthen animal welfare aspects in the DLG-utility testing procedure of housing equipment, which is in place since 1953. The committee elaborates standards concerning testing methods, assessment criteria and the necessary size of investigations. As required, the scientific animal welfare committee may support the DLG-testing bodies in the implementation of the animal welfare part of the testing procedure. It will, moreover, be involved in the welfare assessment based on the testing results. The amendments of the already established testing procedure will help to fulfill the general requirements on an acceptable animal welfare testing procedure. While keeping in mind that there are certain limits in what can be achieved by a voluntary testing procedure, the enhanced consideration of animal welfare aspects within the DLG-utility testing procedure has the advantage to be relatively unbureaucratic and in line with EU legislation, and is, therefore, an appropriate tool for a contibrution to improved animal welfare in livestock housing.

The behavioral endocrinology of domestication: A comparison between the domestic guinea pig (Cavia aperea f. porcellus) and its wild ancestor, the cavy (Cavia aperea).

In this study spontaneous behavior and endocrine parameters were compared between the domestic guinea pig (Cavia aperea f. porcellus) and its wild ancestor, the cavy (Cavia aperea), to elucidate the process of domestication in this species. In 120 h of observation time the behavior of five groups of wild and seven groups of domestic guinea pigs, each consisting of one adult male and two adult females, was analyzed quantitatively. To assess the activities of the pituitary-adrenocortical (PAC), the pituitary-gonadal (PG), and the sympathetic-adrenomedullary (SAM) systems, serum cortisol, testosterone, epinephrine, and norepinephrine concentrations, as well as adrenal tyrosine hydroxylase activities, were determined in males of both forms. The following significant differences between wild cavies and domestic guinea pigs were found: the domesticated animals displayed less aggressive but more sociopositive and more male courtship behavior than their wild ancestors. In addition, they were distinctly less attentive to their physical environment than the wild cavies. The basal activity of the SAM system, as well as the reactivity of the SAM and the PAC systems, was distinctly reduced in the domesticated animals. In contrast, the basal activity of the PAC system did not differ between both forms. The activity of the PG system was significantly higher in males of the domestic guinea pig than in male wild cavies. Thus, in guinea pigs the process of domestication has led to typical behavioral traits-reduced aggressiveness, increased social tolerance-which have also been found in comparisons between wild and domestic forms of other species. The decreased reactivity of the organism's stress axes can be regarded as a physiological mechanism which helps domesticated animals to adjust to man-made housing conditions.

Of domestic and wild guinea pigs: studies in sociophysiology, domestication, and social evolution.

Among mammals a majority of each individual's daily expectations, motivations, and behaviors are directed to encounters with conspecifics. Therefore the knowledge of the genesis, control, and consequences of social interactions is crucial for understanding their social life. We present here our research on the sociophysiology, domestication, and social evolution of wild (Cavia aperea and Galea musteloides) and domestic (Cavia aperea f. porcellus) guinea pigs, which summarizes general rules for many group-living mammals. It is shown that social interactions have consequences not only for the individuals' reproductive success but also for their degrees of stress and welfare. The way in which individuals interact is controlled not only by the present environment but also by the previous social experiences which they have gathered during their behavioral development. Furthermore, the study of ontogeny does not begin at birth, because prenatal social factors acting on pregnant females can also affect the way in which the offspring will interact when adult. In addition, to understand the genesis of interactions between domesticated animals implies knowledge of the behavioral and physiological changes which occurred during the process of domestication. Finally, understanding the social interactions among individuals of the wild ancestor of the domesticated form requires knowledge of how their behavior patterns were brought about by natural selection during the process of social evolution.

The social environment during pregnancy and lactation affects the female offsprings' endocrine status and behaviour in guinea pigs.

This study investigated the influence of the social environment during pregnancy and lactation on the female offsprings' endocrine parameters and behaviour. The subjects studied were daughters whose mothers had either lived in a stable social environment (SSE) during pregnancy and lactation (SE-mothers) or in an unstable social environment (USE) during this period of life (UE-mothers). The SSE was made by keeping the group composition (one male, five females) constant; in the USE situation every third day two females from different groups were exchanged. After weaning, 3 groups of daughters from UE-mothers (UE-daughters) and 3 groups of daughters from SE-mothers (SE-daughters), consisting of four females each, were established. From their 60th through their 80th day of age the spontaneous behaviour of the daughters was recorded in their home cages. On the endocrine level, cortisol and testosterone (T) concentration in serum as well as adrenal tyrosinehydroxylase activities in both adrenals and their weight were determined. UE-daughters displayed significantly higher amounts of male-typical courtship and play behaviour than SE-daughters. This behavioural masculinization corresponded with significantly higher T in UE- than SE-daughters. Cortisol levels did not differ between both categories of females. Significantly higher tyrosinehydroxylase activities and adrenal weight in UE- than SE-daughters pointed, however, to higher degrees of stress response in UE-daughters. Thus, the instability of the social environment during pregnancy and lactation has distinct effects on the behaviour and endocrine system of the female offspring.

Social relationships and the management of stress.

Two different types of social relationships exist in mammalian social systems: dominance relationships and social bondings. This article shows that both are crucial for the management of stress. The following general conclusions are derived: (1) In stable social systems, established dominance relationships result in predictable behaviour. As a consequence, low positions in the hierarchy do not necessarily lead to enhanced endocrine stress responses. Under conditions of instability, however, distinct increases in the activities of the pituitary-adrenocortical- and the sympathetic-adrenomedullary systems are found; (2) The ability to establish and to respect dominance relationships is a prerequisite to build up stable social systems. Whether this ability is realized, however, depends on social experiences made during behavioural development. The time around puberty seems to be essential for the acquisition of those social skills needed to adapt to unfamiliar conspecifics in a non-stressful and non-aggressive way; (3) Stress responses can be ameliorated by the presence of members of the same species. This phenomenon is called social support. In general, social support cannot be provided by any conspecific, but the ability to give social support is restricted to bonding partners. In most mammalian species mothers are important bonding partners for their infants. In some species bondings also occur between adult individuals; and (4) On a physiological level the bonding partner reduces the activities of the pituitary-adrenocortical- and the sympathetic-adrenomedullary systems. On a psychological level he/she can be regarded as a 'security-giving and arousal-reducing structure'. This is true irrespective of whether the bonding partner is the mother, in the case of an infant, or a male or a female in the case of an adult individual.

The social environment, behaviour and stress--a case study in guinea pigs.

In this article we summarize our work on the relationships between the social environment, behaviour and stress in guinea pigs. We confirm what Jim Henry predicted to be a general rule for all mammals including man 20 years ago: The individual's degree of social stress is related to the stability of the pre- and postnatal social environment in which it lives, to the amount of social support which it receives from bonding partners and to the social experiences which it has made during behavioural development.

Prenatal social stress masculinizes the females' behaviour in guinea pigs.

This study investigated the influence of the social environment during pregnancy and lactation on the female offsprings' behaviour and body weight. The subjects studied were daughters whose mothers had lived: a) in a stable social environment (SSE) during pregnancy and lactation; b) in an unstable social environment (USE) during pregnancy and lactation; c) in a SSE during pregnancy but in an USE during lactation; d) in an USE during pregnancy but in a SSE during lactation. The SSE was made by keeping the group composition (one male, five females) constant; in the USE situation every third day two females from different groups were exchanged. After weaning (n = 4 x 3 = 12), groups of daughters, consisting of four females each, were established, originating from the different categories of mothers. From their 41st through their 80th day of age the spontaneous behaviour of the daughters was recorded in their home cages. When the mothers had lived in an USE during pregnancy, the daughters displayed significantly higher amounts of male-typical courtship, play, and social orientation behaviour compared to daughters whose mothers had lived in a SSE during pregnancy. The stability and instability, respectively, of the social environment during lactation, however, had no significant influence on the daughters' prospective behaviour. The four categories of daughters did not differ in their body weights either at birth or at 80 days of age. Thus, the instability of the social environment during pregnancy obviously represented a mild stressor that nevertheless caused a distinct masculinization of the daughters' behaviour.

The environment, hormones, and aggressive behaviour: a 5-year-study in guinea pigs.

This paper summarizes persistent consequences of varying rearing and testing conditions on intermale aggression, endocrine responses, and body weight (BW) in guinea pigs. Pairs of unfamiliar males were placed in chronic confrontations for 3-50 days in 2 m2 enclosures. Their behaviour was recorded in more than 1000 h of observation time. About 900 samples were assayed for plasma glucocorticoid (GC), testosterone (T), and norepinephrine (NE) before and at different times after the beginning of the confrontations. The males were reared either in large mixed-sex colonies (CRM) or with a single female (FRM). When FRM males were confronted, high levels of aggressive behaviour occurred and high degrees of stress were found in losers, especially when the environment was familiar and a female was present. In contrast, CRM males confronted in the same situation, but in a nonaggressive way, showed no changes in GC, NE, and BW. These differences between FRM and CRM developed around puberty. A 50-min agonistic experience with an unfamiliar male around this time shifted the subsequent adult behaviour of a FRM to a CRM pattern. These findings suggest a causal relationship between social experiences occurring around puberty, subsequent behaviour as adults, and degree of stress in chronic social encounters.

The ability to arrange with conspecifics depends on social experiences around puberty.

The study examined the effect of age on the ability to arrange with unfamiliar conspecifics in differentially reared male guinea pigs. The experiments were conducted with 50-, 90-, 150-, and 210-240-day-old-males, which either had been reared in large colonies (CRM) or each with a single female (FRM). In each age class, seven pairs of unfamiliar CRM and seven pairs of unfamiliar FRM were placed in chronic confrontations for 6-10 days in the presence of an unfamiliar female in 2 m2 enclosures. Body weights were determined directly before and 52 h, 124 h, and 220 h after the onset of the experiments. CRM showed no substantial losses in body weights irrespective of whether the experiments were conducted before, around, or after sexual maturity (which is achieved at 2-3 months of age). In contrast pairs of FRM arranged without problems only at 50 days of age. Around puberty, social stress distinctly increased, and 2 and 4-5 months later the confrontations even had to be ceased after 6 days, because losers extremely decreased in body weights. It is concluded that in male guinea pigs the time around sexual maturity is crucial for the development of social skills necessary to arrange with unfamiliar conspecifics in a nonstressful way.

Social experience, behavior, and stress in guinea pigs.

This study examined the effect of varying rearing and testing conditions on guinea pig aggression, courting behavior, endocrine responses and body weight. Pairs of 7-8-month-old males were placed in chronic confrontations for 6-50 days in 2 m2 enclosures. Social behavior was recorded with a total of 882 h observation time. Body weight as well as plasma glucocorticoid, testosterone and norepinephrine titers were determined for each male 20 h before, and 4, 52 and 124 h after, the onset of the chronic encounters. Three experiments were conducted: in Experiment I, 7 pairs of males, each male raised singly with one female (FRM), were confronted in the presence of an unfamiliar female, in Experiment II, 6 pairs of FRM were confronted with no female present, and in Experiment III, 7 pairs of males which were raised in different large colonies were confronted in the presence of an unfamiliar female. In Experiment II and III low levels of aggression, no distinct endocrine changes and no indications of physical injury occurred in winners or losers, whereas in Experiment I high levels of aggression and courting behavior, extreme increases in glucocorticoid titers and distinct decreases in body weights were found in both males. Losers, however, were affected to a much greater extent than winners. These findings suggest that in guinea pigs a causal relationship exists between social rearing conditions, behavior as adults and degree of social stress in chronic encounters.