Stéphane Andanson

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.

The motivation of cows to walk as thwarted by tethering

Cows are often housed in tie-sheds for some part of the year (from 20 to 80% of adult cows in European countries). By contrast, regulations aiming at improving animal welfare generally provide for more opportunity to express behavior and so limit tethering (the European directive for sows). To inform current debate on whether cows should be tethered from an animal welfare point of view, 2 experiments were conducted to determine whether cows were motivated to walk (if their walking motivation was thwarted by tethering) and whether this frustration resulted in acute or chronic stress. In Exp. 1, fifteen cows were housed in tie-stalls for 1, 3, 9, or 27 d or loose-housed in a pen equipped with free-stalls for 27 d according to a Latin square design. On d 28, cows were observed in an 80-m(2) arena where they could walk or run for 10 min. In Exp. 2, fifteen cows were either loose-housed or tie-housed with or without 1 h of daily access to an exercise area according to a Latin square design (treatment duration of 27 d). On d 28, cows were observed in the test arena for 10 min as in Exp. 1. In addition, in Exp. 2, milk was sampled twice weekly for cortisol concentrations and cows underwent a chronic stress test (challenge with ACTH, followed by blood samples for cortisol concentrations). When they were tied with no access to exercise, cows displayed a greater locomotor activity in the test arena (time spent walking x 1.4 in Exp. 1, P < 0.05; time spent trotting x 5 in Exp. 2, P < 0.05), whatever the duration of tie-housing. Regular exercise caused locomotor activity in the arena test to revert to levels observed when cows were loose-housed. Basal cortisol concentrations in milk decreased with time whatever the housing condition (P < 0.001). This decrease was more marked when cows were tied than when they were loose-housed (P < 0.05). Cortisol responses to ACTH were similar between treatments. Adult cows are motivated to walk, and tethering thwarts this motivation. However, the frustration produced by tethering does not result in either acute or chronic physiological stress responses. We recommend that cows housed in tie-sheds be given regular access to an exercise area.

Effects of feed deprivation on behavioral reactivity and physiological status in Holstein cattle.

The present study evaluated whether feed deprivation can increase reactivity to stressful events, such as those that can occur at slaughter. Therefore, effects of 30 h of feed deprivation on behavior, including reactions to psychological stressors, and physiological status in cattle were determined. Sixteen Holstein cows (Exp. 1) and 32 Holstein heifers (Exp. 2) were either fed (FE) or 30-h feed deprived (FD). Throughout the first day of feed deprivation and during evening feed distribution to control animals, FD heifers and cows were more active than controls (P < 0.05). In Exp. 1, during a feeding test, in response to a sudden air blast arising from the bucket from which the cow was feeding, FD cows showed a longer latency to return to feed (P = 0.0002), spent less time in the bucket air blast zone (P = 0.008) and less time motionless (P = 0.03), and tended to withdraw over a longer distance (P = 0.07) than FE cows. In Exp. 2, during a reactivity test, FD heifers spent more (P = 0.0001) time motionless in response to social isolation than FE heifers. In Exp. 2, one-half of the FE and FD heifers were subjected to an additional physical and psychological stressor just before the reactivity test by driving them for 5 min through a labyrinth. Within heifers subjected to the additional stressor, FD heifers were less accepting of being detained (P = 0.05) and stroked (P = 0.003) by a familiar stockperson in a corner of the test arena. Compared with FE animals, FD heifers and FD cows had greater plasma cortisol concentrations (P < 0.05). Feed-deprived cows also had reduced β-hydroxybutyrate concentrations (P = 0.02) compared with FE cows. Thus, in cattle, FD influenced some of the classical indicators of energy metabolism and exacerbated reactivity to sudden events. In addition, when additional stressors were applied, FD cattle were more reluctant to accept handling. Results indicate that a multifactorial origin of stressors during the slaughter period may synergistically increase psychological stress of cattle.

An assessment of the impact of rumenocentesis on pain and stress in cattle and the effect of local anaesthesia

Rumenocentesis is commonly used to collect rumen fluid to screen for acidosis. This study was designed to investigate whether rumenocentesis induced pain and stress and, if so, whether local anaesthesia could limit this. Twenty-four dairy cows were assigned to one of three treatments: (1) rumenocentesis with local anaesthesia (AR); (2) rumenocentesis without local anaesthesia (R); and (3) local anaesthesia only (A). Treatments were performed in a restraining cage. The cows were placed in the cage on three consecutive days and anaesthesia and/or rumenocentesis was performed on the second day. Blood samples for cortisol determination and heart rate were recorded from 0.25 h before treatment until 4h after. Behaviour was noted while the cows were caged. Feed intake and milk production were measured the week before treatment, on the day of treatment, and the day after. With all three treatments, cortisol concentrations and heart rate were increased while cows were in the cage. Cortisol, cardiac and behavioural responses were not significantly higher in the R and AR treatments than the A group. Cortisol concentrations and heart rate did not change between days. Feed intake and milk production were unaffected by the treatments. It was concluded that rumenocentesis does not appear more stressful than local anaesthesia or handling.

Stress during pregnancy alters dendritic spine density and gene expression in the brain of new-born lambs.

Rodent studies show how prenatal stress (PS) can alter morphology in the cortico-limbic structures that support emotional and cognitive functions. PS-induced alteration is less well described in species with a gyrencephalic brain and complex earlier fetal development, and never in sheep at birth to rule out postnatal environment effects or influences of maternal behavior. This study aimed to assess the consequences of a mild chronic stress in pregnant ewes on the neurobiological development of their lambs at birth. During the last third of gestation, 7 ewes were exposed daily to various unpredictable and negative routine management-based challenges (stressed group), while 7 other ewes were housed without any additional perturbation (control group). For each group, a newborn from each litter was sacrificed at birth to collect its brain and analyze its expression levels of genes involved in neuronal dendritic morphology (Dlg4, Rac1, RhoA, Doc2b), synaptic transmission (Nr1, Grin2A, Grin2B) and glucocorticoid receptor (Nr3C1) in hippocampus (HPC), prefrontal cortex (PFC) and amygdala (AMYG). Results revealed that lambs from stressed dam (PS lambs) showed under-expression of Rac1 and Nr1 in PFC and overexpression of Dlg4 in AMYG compared to controls. To assess the morphological consequences of gene dysregulations, the dendritic morphology of pyramidal neurons was explored by Golgi-Cox staining in HPC and PFC. PS lambs had higher dendritic spine density in both structures and more stubby-type spines in the CA1 area of HPC than controls. This is the first demonstration in sheep that PS alters fetal brain, possibly reflecting functional changes in synaptic transmission to cope with adversity experienced in fetal life.

Heat stress in cows at pasture and benefit of shade in a temperate climate region

Under temperate climates, cattle are often at pasture in summer and are not necessarily provided with shade. We aimed at evaluating in a temperate region (Belgium) to what extent cattle may suffer from heat stress (measured through body temperature, respiration rate and panting score, cortisol or its metabolites in milk, and feces on hot days) and at assessing the potential benefits of shade. During the summer of 2012, 20 cows were kept on pasture without access to shade. During the summer of 2011, ten cows had access to shade (young trees with shade cloth hung between them), whereas ten cows had no access. Climatic conditions were quantified by the Heat Load Index (HLI). In animals without access to shade respiration rates, panting scores, rectal temperatures, and milk cortisol concentrations increased as HLI increased in both 2011 and 2012. Fecal cortisol metabolites varied with HLI in 2011 only. When cattle had access to shade, their use of shade increased as the HLI increased. This effect was more pronounced during the last part of the summer, possibly due to better acquaintance with the shade construction. In this case, shade use increased to 65% at the highest HLI (79). Shade tempered the effects on respiration, rectal temperature, and fecal cortisol metabolites. Milk cortisol was not influenced by HLI for cows using shade for > 10% of the day. Therefore, even in temperate areas, cattle may suffer from heat when they are at pasture in summer and providing shade can reduce such stress.