Birger Puppe

Consequences of repeated early isolation in domestic piglets (Sus scrofa) on their behavioural, neuroendocrine, and immunological responses.

Stress in the form of intermittent maternal deprivation and social isolation during early postnatal life in rats and monkeys produces persistent changes in physiology and behaviour. In farm animals physiological consequences of disrupting mother-infant interactions with respect to health and animal welfare are relatively unknown. Therefore, the aim of the present study was to investigate the behavioural, neuroendocrine and immunological consequences of a 2 h daily social isolation from day 3 to day 11 of age in domestic piglets as well as potential long-term effects on the brain-endocrine-immune regulation. Repeated social isolation resulted in significantly decreased open-field activity (locomotion, vocalization) during the isolation period, increased basal cortisol concentrations and decreased lymphocyte proliferation in response to concanavalin A and pokeweed mitogen one day after the isolation. There was also a significant increase of interleukin-1beta (IL-1beta) concentration in hippocampus in isolated piglets compared to controls at this time. Six weeks after isolation significant enhanced basal ACTH concentrations as well as higher IL-1beta content and glucocorticoid receptor (GR) binding in hippocampus were found. These endocrine and immune responses were associated with decreased CRH levels in the hypothalamus and increased CRH content in the amygdala. The present data indicate that early social isolation in pigs may cause changes in behavioural, neuroendocrine, and immune regulation and produce long-term effects not only on the activity of the hypothalamic-pituitary-adrenal (HPA) system, but also on the immune-brain circuitry with possible negative consequences in health and welfare of commercial pigs. Using the pig as a suitable animal model, the finding of this study may also have some implications for the etiology of anxiety and depression in humans.

Effects of social status after mixing on immune, metabolic, and endocrine responses in pigs

The effects of social rank on immune, metabolic, and endocrine responses were studied in 10 newly mixed groups of German Landrace pigs (9 individuals each) at an age of 12 weeks. Immediately after mixing, the agonistic interactions (AI) of all group members were continuously recorded over 3 days (10 h daily). An individual dominance value (DV) was calculated by the number of wins minus defeats in relation to all decisive fights (DV < or = 0, subordinate; DV > 0, dominant). Blood samples were taken 24 h before and 3 days after mixing. The data showed that the social status had a significant effect on lymphocyte proliferation in responses to different mitogens: socially dominant pigs had higher proliferative response than subordinate pigs. In addition, during the observation period the lymphocyte activation by mitogens increased in the dominant animals and decreased in the subordinate animals with increasing number of agonistic interactions. The rise in total serum IgG concentration 3 days after mixing was higher in dominant pigs compared with subordinates. The dominance status did not significantly affect plasma metabolic levels nor cortisol concentrations. However, mixing appeared to increase glucose and total protein values and to decrease alkaline phosphatase and cortisol levels in both, dominant and subordinate pigs. In conclusion, mitogen induced cell proliferation seems to be a valuable marker for acute social stress in pigs.

Effects of familiarity and relatedness on agonistic pair relationships in newly mixed domestic pigs

Abstract The effects of familiarity and relatedness on agonistic pair relationships (dyads) in different pen regions (pen area or trough area) were studied in 16 groups of newly mixed domestic pigs of similar weight (9 pigs per group) at an age of 12 weeks. The agonistic interactions (AI) within 124 familiar (related) and 452 unfamiliar (related and unrelated) dyads were continuously recorded for 3 days (10 h daily) after mixing. Whereas pigs, both in dyads familiar and unfamiliar to each other, showed the same frequency of AI in the trough area, unfamiliar dyads exhibited significantly more AI in the pen area than familiar dyads. The relatedness of unfamiliar dyads had apparently no influence on AI. It is discussed that, besides establishing a dominance hierarchy, pigs react aggressively on strange subjects. Furthermore, the results are briefly discussed with reference to dominance and resource usage in pigs.

Effects of postnatal social isolation on hormonal and immune responses of pigs to an acute endotoxin challenge

Social stress during early postnatal life often results in long-term effects on neuroendocrine and immune adaptation mechanisms. Therefore, the aim of the present study was to determine the influence of a 2-h daily social isolation from Day 3 to Day 11 on the acute and long-term proinflammatory and neuroendocrine responses of piglets challenged with the bacterial endotoxin lipopolysaccharide (LPS; 100 microg/kg body weight). Peripheral LPS administration significantly increased plasma concentrations of tumor necrosis factor-alpha (TNF-alpha), ACTH and cortisol in isolated and control pigs. However, the activity of the hypothalamic-pituitary-adrenal (HPA) axis after LPS stimulation was not significantly affected by isolation treatment, whereas the prior social isolation diminished the plasma TNF-alpha response to LPS 1 day as well as 45 days after the isolation period. The hippocampal TNF-alpha concentration in response to LPS was also reduced in priorly isolated pigs compared to control animals. Furthermore, the significant increase of TNF-alpha in the spleen caused by LPS was associated with a dramatic decrease in glucocorticoid receptor (GR) binding. The GR binding in hippocampus was increased in isolated pigs and was significantly decreased after LPS injection. In addition, the repeated isolation stressor was shown to increase hippocampal levels of interleukin-1beta (IL-1beta). The present results indicate that repeated social isolation of neonatal pigs may cause long-term effects on proinflammatory regulation at the periphery and in the brain following immune challenge with particular importance of TNF-alpha in mediating these interactions.

Agonistic interactions and physiological reactions of top- and bottom-ranking pigs confronted with a familiar and an unfamiliar group: Preliminary results

Abstract Four trials were carried out, where in each trial during a selection test two groups of growing pigs (12 weeks old) with nine animals each were randomly formed. After a week, the top (TR) and bottom (BR) ranking pig from each group was isolated and kept under single housing conditions. At the age of 15 to 16 weeks, the test animals were confronted once with the familiar and once with the unfamiliar group. During the 10-h social confrontation test, agonistic behaviour (agonistic interactions, individual dominance values) and physiological stress reactions (epinephrine, norepinephrine, cortisol, heart rate) were investigated simultaneously. The social confrontation of a pig with a familiar or an unfamiliar group caused very frequent agonistic interactions during the first 30 min. This behaviour was accompanied by a rapid increase of the heart rate and the plasma catecholamine concentrations and a delayed increase of cortisol. TR pigs elicited more agonistic interactions during the first 30 min and displayed higher plasma catecholamine concentrations. Additionally, TR animals showed significantly higher dominance values during the 10-h testing situation compared to the BR pigs. The confrontation of an animal with the unfamiliar group resulted in a more pronounced adrenocortical reaction compared to the confrontation with the familiar group. In particular, the TR animals showed a distinct increase of plasma cortisol during the first hour and a later bradycardia when they were confronted with the unfamiliar group. The preliminary results presented in this paper indicate that pigs show different, but inconsistent behavioural and physiological reactions when they are confronted with a social group. The agonistic behaviour and the physiological stress reactions during a social confrontation test are still influenced by the former rank of the animal, even when the animal was isolated under single housing conditions in the meantime for 2 to 3 weeks.

Autonomic reactions indicating positive affect during acoustic reward learning in domestic pigs

Cognitive processes, such as stimulus appraisal, are important in generating emotional states and successful coping with cognitive challenges is thought to induce positive emotions. We investigated learning behaviour and autonomic reactions, including heart rate (HR) and its variability (standard deviation (SDNN) and root mean square of successive differences (RMSSD) of a time series of interbeat intervals). Twenty-four domestic pigs, Sus scrofa, housed in six groups of four, were confronted with a cognitive challenge integrated into their familiar housing environment. Pigs were rewarded with food after they mastered the discrimination of an individual acoustic signal followed by an operant task. All pigs quickly learned the tasks, while baseline SDNN and RMSSD increased significantly throughout the experiment. In reaction to the signals, pigs showed a sudden increase in HR, SDNN and RMSSD, and a decrease in the RMSSD/SDNN ratio. Immediately after this reaction, the HR and SDNN decreased, and the RMSSD/SDNN ratio increased. During feeding, the HR and the RMSSD/SDNN ratio stayed elevated. The pigs showed no cardiac reaction to the sound signals for other pigs or their feeding pen mates. We concluded that the level of cognitive challenge was adequate and that the observed changes in the autonomic tone, which are related to different dimensions of the affective response (e.g. arousal and valence), indicated arousal and positive affective appraisal by the pigs. These findings provide valuable insight into the assessment of positive emotions in animals and support the use of an adequate cognitive enrichment to improve animal welfare.

A single exposure to social isolation in domestic piglets activates behavioural arousal, neuroendocrine stress hormones, and stress-related gene expression in the brain

Stressful early life events can have short- and long-term effects on neuroendocrine and behavioural mechanisms of adaptation. Here, we investigated the effects of a single social isolation (4 h) of domestic piglets on both behavioural alterations in open-field tests and modifications in the expression of genes regulating glucocorticoid response in stress-related brain regions at 7, 21 or 35 days of age. The mRNAs of glucocorticoid receptor (GR), mineralocorticoid receptor (MR), 11ss-hydroxysteroid dehydrogenase 1 and 2 (11ss-HSD1 and 11ss-HSD2) and c-fos were analysed by real-time RT-PCR in the hypothalamus, hippocampus and amygdala. The social isolation caused both elevated stress hormone concentrations (e.g. cortisol) and open-field reactivity (e.g. locomotion, vocalisation) compared to control piglets. The enhanced behavioural and neuroendocrine activity was associated with distinct changes in gene expression in the limbic system. The hypothalamic GR, MR and 11ss-HSD1 mRNA expressions and the hippocampal 11ss-HSD1 mRNA was significantly higher in isolated piglets, whereas in the amygdala social isolation caused a significant decrease in MR mRNA expression. Isolated piglets also displayed significantly higher c-fos mRNA expression, an estimate of neuronal activation, in hypothalamus and amygdala. The mRNA alterations as well as the behavioural and hormonal pattern show an effect of social isolation on days 7 and 21, but no effect on day 35. In conclusion, a single social isolation in piglets caused age-dependent neuroendocrine and behavioural changes that indicate increased arousal and experienced distress. The present results also suggest that psychosocial stress effects should be considered for the assessment of livestock handling practices with respect to health and welfare.

Linear prediction coding analysis and self-organizing feature map as tools to classify stress calls of domestic pigs (Sus scrofa)

It is assumed that calls may give information about the inner (emotional) state of an animal. Hence, in the last years sound analysis has become an increasingly important tool for the interpretation of the behavior, the health condition, and the well-being of animals. A procedure was developed that allows the characterization, classification, and visualization of the cluster structures of stress calls of domestic pigs (Sus scrofa). Based on the acoustic model of the sound production the extraction of features from calls was performed with linear prediction coding (LPC). A vector-based self-organizing neuronal network was trained with the determined LPC coefficients, resulting in a feature map. The cluster structure of the calls was then visualized with a unified matrix and the neurons were labeled for their input origin. The basic applicability of the procedure was tested by using two examples which were of special interest for a possible evaluation of the normal farming practice. The procedure worked well both in discriminating individual piglets by their scream characteristics and in classifying pig stress calls vs other calls and noise occurring under normal farming conditions.

Physiological and behavioral effects of different success during social confrontation in pigs with prior dominance experience

The impact of a 10-h social confrontation on behavior, plasma adrenaline, noradrenaline and cortisol was studied in 14 domestic pigs (eight castrated males and six females) with prior dominance experience. Prior to the experiments, animal groups, each consisting of nine growing pigs (12 weeks old) from different litters, were composed randomly. After 5 days, the pig with the highest rank from each group was removed, provided with a jugular vein catheter and kept in single housing for 2-3 weeks. After this period, each test animal was returned into its familiar group for a 10-h social confrontation. The reintroduction of the test animals into the familiar groups caused frequent agonistic interactions during the first 30 min. Seven animals succeeded to win most of their encounters during the confrontation test and were classified as High Success (HS) animals, whereas seven other animals lost most of their encounters and were classified as Low Success (LS) animals. The reintroduction of the test animals into the groups provoked also marked changes in behavioral and physiological measures. The frequent fighting behavior during the first 30 min was accompanied by a rapid increase of plasma catecholamines and a delayed increase of cortisol. Immediately after introduction, LS pigs tended to show higher plasma adrenaline and noradrenaline concentrations than HS pigs. There was also a tendency for a sustained increase of noradrenaline/adrenaline ratios in HS pigs, whereas the ratios of LS pigs remained nearly unchanged. No significant differences between both groups were found in cortisol levels and in the frequency of agonistic interactions. However, LS animals showed less locomotion, more lying and spent less time exploring the pen or other animals. These results show that during a social confrontation the experience of defeats for formerly high-ranking pigs is accompanied by increased submissive or passive behavior and a higher sympathoneural and adrenomedullary reaction, which may indicate more emotional distress and fear compared to successful animals.

Effects of attention and rewarded activity on immune parameters and wound healing in pigs.

Challenging animals with a demanding situation they are able to cope with and where they are rewarded may be a source of positive emotions and possibly of increased welfare. In order to test if this results in changes of immunological parameters and wound healing, 56 pigs (7-20 weeks of age), housed in groups of 8 animals each, were successfully trained to recognize and localize an individual acoustic summons and to receive a small portion of feed as a reward. Immune reactions and the development of a standardized biopsy wound were compared to values of conventionally fed control groups of equal size and animal-to-feeding-place ratio (2:1). In the experimental animals a significantly higher concentration of IgG as well as an increased in vitro T-cell proliferation to ConA but a reduced LPS-induced proliferation of B-cells was found, while basal salivary cortisol concentrations were similar. Wound development was better in the experimental animals as measured by the area of the inflammatory corona. It decreased more rapidly in the experimental animals since the 5th day after biopsy and was significantly smaller than in the control groups. We conclude that environmental enrichment by equipment provoking attention and cognitive activity which is rewarded by feed may play a beneficial role for physical welfare of intensively housed pigs.