Cornelis G. van Reenen

Exploration of the hypothalamic–pituitary–adrenal function as a tool to evaluate animal welfare

Measuring HPA axis activity is the standard approach to the study of stress and welfare in farm animals. Although the reference technique is the use of blood plasma to measure glucocorticoid hormones (cortisol or corticosterone), several alternative methods such as the measurement of corticosteroids in saliva, urine or faeces have been developed to overcome the stress induced by blood sampling itself. In chronic stress situations, as is frequently the case in studies about farm animal welfare, hormonal secretions are usually unchanged but dynamic testing allows the demonstration of functional changes at several levels of the system, including the sensitization of the adrenal cortex to ACTH and the resistance of the axis to feedback inhibition by corticosteroids (dexamethasone suppression test). Beyond these procedural aspects, the main pitfall in the use of HPA axis activity is in the interpretation of experimental data. The large variability of the system has to be taken into consideration, since corticosteroid hormone secretion is usually pulsatile, follows diurnal and seasonal rhythms, is influenced by feed intake and environmental factors such as temperature and humidity, age and physiological state, just to cite the main sources of variation. The corresponding changes reflect the important role of glucocorticoid hormones in a number of basic physiological processes such as energy metabolism and central nervous system functioning. Furthermore, large differences have been found across species, breeds and individuals, which reflect the contribution of genetic factors and environmental influences, especially during development, in HPA axis functioning. Usually, these results will be integrated with data from behavioral observation, production and pathology records in a comprehensive approach of farm animal welfare.

Adrenocortical reactivity and central serotonin and dopamine turnover in young chicks from a high and low feather-pecking line of laying hens.

Feather pecking in domestic fowl is a behavioral abnormality that consists of mild or injurious pecking at feathers of conspecifics. Previously, it was shown that chicks from a high feather-pecking (HFP) and low feather-pecking (LFP) line of laying hens already differ in their propensity to feather peck at 14 and 28 days of age. As a first step in investigating a possible relationship between the development of feather pecking and physiological and neurobiological characteristics of laying hens, two subsequent experiments were carried out. Firstly, we investigated the development of adrenocortical (re)activity in HFP and LFP chicks during the first 8 weeks of life. Secondly, we studied dopamine (DA) and serotonin (5-HT) turnover in the brain of 28-day-old HFP and LFP chicks. In both experiments, chicks were exposed to manual restraint (placing the chicks on its side for 5 min). Plasma corticosterone levels were lower (baseline on Days 3 and 56; restraint-induced on Days 3, 14 and 28) in HFP chicks. Both brain DA and 5-HT turnover were lower in the HFP chicks, as well. Possible consequences for the observed differences in (stress) physiology and neurobiology between the two lines in relation to the feather pecking are discussed.

The development of feather pecking behaviour and targeting of pecking in chicks from a high and low feather pecking line of laying hens

Large individual differences between adult laying hens in their propensity for feather pecking are known to exist. However, not much research has been carried out into the individual differences concerning the development of feather pecking behaviour. The purpose of this study was to investigate whether contrasting levels of feather pecking, observed among adult birds from two lines of laying hens, already occur at an early age. Further-more, an important question to be discussed was whether different behavioural systems may be related to the occurrence of feather pecking. Therefore, this study consisted of studying and comparing the behaviour of White Leghorn laying hens from a high (HFP) and low feather pecking line (LFP) during the first 8 weeks of life. Chicks were reared in litter-floor pens and were kept in groups of five animals per line (12 groups per line). HFP chicks showed significantly higher levels of gentle feather pecking (gentle FP) than LFP chicks at the age of 14 and 28 days. Furthermore, HFP chicks spent significantly more time preening than LFP chicks on days 14, 28 and 41. Duration of foraging behaviour and feeding behaviour was significantly higher in the LFP line compared to the HFP line on days 41 and 56 and days 28, 41 and 56, respectively. HFP chicks showed a significant negative correlation between gentle FP and preening on days 3 (r = -0.49) and 41 (r = -0.86). In the LFP line duration of feeding correlated negatively with gentle FP on day 3 (r = -0.63). A principal component analysis (PCA) revealed that in the HFP line, gentle FP and preening exhibited high and opposite loadings on the same component at all ages, whereas feeding consistently loaded on the other component. This outcome contrasted with that of the LFP line. In this line feeding predominantly loaded on the same principal component as gentle FP with loadings opposite to those of gentle FP, whereas preening showed the same loadings as gentle FP, on days 3 and 41. In conclusion, differences in feather pecking behaviour between HFP and LFP chicks can already be observed at a very early age during development. Furthermore, our results indicate that HFP and LFP chicks differ in the way pecking behaviour is targeted. This difference could be related to the existence of a difference in underlying motivational system controlling the development of feather pecking between the two lines

Age, gender and litter-related variation in T-lymphocyte cytokine production in young pigs

The capacity of farm animals to produce cytokines could be an important determinant of robustness and health. From research in rodents and humans it appears that the production and the balance of T helper 1 (Th1) and T helper 2 (Th2)‐type cytokines influences susceptibility to autoimmune and infectious diseases. It is known that pigs show a large variation in many immune response parameters. So far the extent of individual variation in the production of Th1‐ and Th2‐type cytokines in commercial outbred pigs has not been reported. In the current experiment we determined mRNA expression, as well as protein production of cytokines in 32 pigs from eight litters. From each litter two male and two female pigs were tested at 2, 5 and 8 weeks of age. Two Th1‐type cytokines, interleukin (IL)‐2 and interferon (IFN)‐gamma, and two Th2‐type cytokines, IL‐4 and IL‐10, were measured after phytohaemagglutinin (PHA)‐stimulation of blood mononuclear cells. Cytokine production and the Th1/Th2‐ratio were highly variable. The variation in cytokine protein production was moderately consistent across ages, i.e. pigs that produced high levels of cytokine at 2 weeks of age tended to do so as well at 5 and 8 weeks of age. Cytokine production tended to increase with age, and gilts and boars differed in their IL‐2/IL‐4 ratio. Unexpectedly, age, gender and litter effects often differed for mRNA and protein production data. We hypothesize that cytokine production is a consistent trait in pigs, especially at the protein production level. Future investigations in more animals and across a wider age range are necessary.

Welfare assessment: correlations and integration between a Qualitative Behavioural Assessment and a clinical/ health protocol applied in veal calves farms

Abstract This study is aimed at finding correlations and possible integration among Qualitative Behavioural Assessment (QBA) and a specific protocol of clinical/health evaluation. Both welfare assessment methods were based on direct animal observation and were applied in 24 Italian veal calves farms at 3 weeks (wks) of rearing. Principal component analysis (PCA) summarized 20 QBA descriptors on two main components (PC1 and PC2) with eigenvalues above 4 and explaining 29.6 and 20.3% of the variation respectively. PCA on residuals obtained after correcting for housing condition yielded highly similar results, indicating that the rearing environment of the calves was not an important determinant of the observer reliability of QBA. A relationship was found between QBA PC2 and the presence of signs of cross-sucking recorded during the clinical visit (presence PC2=1.11 vs. absence PC2=-1.55, P<0.001). There were no other relations between the quantitative clinical measures and QBA PC’s. The frequency of farmer, veterinarian, or industry technician visits to the veal unit significantly affected QBA PC1 and PC2 scores. These results suggest that the 2 methods provide complementary types of information and can each make valid a contribution to an integrated animal welfare monitoring scheme.

Tail Biting in Pigs: Blood Serotonin and Fearfulness as Pieces of the Puzzle?

Tail biting in pigs is a widespread problem in intensive pig farming. The tendency to develop this damaging behaviour has been suggested to relate to serotonergic functioning and personality characteristics of pigs. We investigated whether tail biting in pigs can be associated with blood serotonin and with their behavioural and physiological responses to novelty. Pigs (n = 480) were born in conventional farrowing pens and after weaning at four weeks of age they were either housed barren (B) or in straw-enriched (E) pens. Individual pigs were exposed to a back test and novel environment test before weaning, and after weaning to a novel object (i.e. bucket) test in an unfamiliar arena. A Principal Component Analysis on behaviours during the tests and salivary cortisol (novel object test only) revealed five factors for both housing systems, labeled ‘Early life exploration’, ‘Near bucket’, ‘Cortisol’, ‘Vocalizations & standing alert’, and ‘Back test activity’. Blood samples were taken at 8, 9 and 22 weeks of age to determine blood platelet serotonin. In different phases of life, pigs were classified as tail biter/non-tail biter based on tail biting behaviour, and as victim/non-victim based on tail wounds. A combination of both classifications resulted in four pig types: biters, victims, biter/victims, and neutrals. Generally, only in phases of life during which pigs were classified as tail biters, they seemed to have lower blood platelet serotonin storage and higher blood platelet uptake velocities. Victims also seemed to have lower blood serotonin storage. Additionally, in B housing, tail biters seemed to consistently have lower scores of the factor ‘Near bucket’, possibly indicating a higher fearfulness in tail biters. Further research is needed to elucidate the nature of the relationship between peripheral 5-HT, fearfulness and tail biting, and to develop successful strategies and interventions to prevent and reduce tail biting.