A randomized controlled trial to study the effect of supplemental premilking stimulation on milking performance, teat tissue condition, udder health, and well-being in dairy cows.

The objectives of this study were to evaluate the effect of supplemental premilking stimulation, provided after manual stimulation, by means of high-frequency pulsation without reduction of the vacuum in the pulsation chamber on milking performance, teat tissue condition, udder health, and well-being in dairy cows. In a randomized controlled trial, Holstein cows (n = 491) from 1 commercial dairy farm with a thrice-per-day milking schedule were assigned to treatment and control groups over a 64-d period. Treatments consisted of a maximum of 20 s of pulsation stimulation at a pulsation rate of 100 (SPS100) or 300 (SPS300) cycles per minute and a pulsation ratio of 25:75, which were applied after completion of manual premilking stimulation upon milking unit attachment. Cows in the control group (CON) received only traditional premilking stimulation by manual forestripping for 6 s, and wiping. Milking characteristics were measured with on-farm milk flow meters. Milking machine-induced short-term (swelling at teat base, firmness at teat end, and teat discoloration) and long-term (teat-end callosity) changes to the teat tissue were assessed manually and visually. Composite milk samples were analyzed for somatic cell counts. Cow hind-leg activity was assessed with 3-dimensional accelerometers. Fecal 11,17-dioxoandrostanes (11,17-DOAs; a group of cortisol metabolites) were determined on wk 4 and 8 to assess the well-being of the cows. Generalized linear mixed models were used to study the effect of treatments on the outcome variables milk yield/milking and milking unit-on time. We observed no meaningful differences among groups for milk yield or milking unit-on time. Least squares means and their 95% confidence intervals (95% CI) for cows in the SPS100, SPS300, and CON groups were 13.9 (13.3-14.5), 14.0 (13.4-14.6), and 13.9 (13.3-14.6) kg for milk yield and 218 (212-224), 218 (211-224), and 218 (212-224) s for milking unit-on time, respectively. Compared with cows in the CON group, the odds (95% CI) of short-term changes were 1.30 (0.95-1.78) for the SPS100 group and 1.50 (1.10-2.05) for the SPS300 group. The odds of long-term changes were 0.94 (0.67-1.34) for cows in the SPS100 group and 0.71 (0.49-1.04) for cows in the SPS300 group. We observed no differences in SCC. In reference to the CON group, the hazard ratio (95% CI) in SPS100 and SPS300, respectively, were 0.35 (0.13-0.98) and 1.22 (0.57-2.64) for clinical mastitis, and 0.34 (0.12-0.95) and 1.28 (0.60-2.73) for culling. The least squares means (95% CI) of hind-leg activity during milking were 8.3 (6.5-10.5), 10.6 (8.1-13.7), and 9.1 (7.2-11.6) movements per milking for the SPS100, SPS300, and CON groups, respectively. The least squares means (95% CI) of fecal 11,17-DOAs concentration (ng/g) at the first and second test days, respectively, were 31.1 (28.1-34.2) and 22.3 (19.2-25.4) for the SPS100 group, 26.4 (23.4-29.4) and 25.2 (22.0-28.4) for the SPS300 group, and 24.8 (21.8-27.9) and 25.0 (21.7-28.3) for the CON group. We conclude that applying supplemental stimulation after manual stimulation through the high-frequency pulsation system tested here did not impart additional benefits to the milk harvesting process, teat tissue condition, somatic cell count, or the well-being of the animals.

Management of dairy heifers: Can operant conditioning be an effective and feasible tool to decrease stress and ease animals' close contact and handling?

Besides health monitoring, a regular check of dairy heifers' growth rate is desirable, but it is rarely done because procedures that require restraint and handling can be associated with substantial stress for both animals and farmers. Inexperienced heifers, especially if they are highly responsive to humans, may find restraint and handling potentially aversive. This study investigated whether training heifers of different age and responsiveness toward humans (RTH), through operant conditioning, could reduce stress in animals, ease close contact and handling, and be feasible in terms of farmers' effort. We assessed 60 Holstein heifers of 2 age classes (young, n = 29, 291 ± 39 d; old, n = 31, 346 ± 62 d) according to the avoidance distance test and classified them as confident (n = 20), neutral (n = 21), or nonconfident (n = 19). Half of the heifers of each age and RTH class were trained (n = 29), whereas the other half was not (n = 31). The trained heifers were subjected to target training for 8 sessions and positively reinforced with feed to allow being touched on the muzzle, rump, and perineum. If a heifer refused positive reinforcement, the trainer stepped back as negative reinforcement. In the last week of the experiment, the effect of training on the reaction to handling was assessed in all heifers. We measured heart rate, root mean square of successive interbeat interval differences (RMSSD), and fecal cortisol metabolites (FCMet). The presence of behavioral distress signs was recorded as well. The avoidance distance test was performed a second time 24 h after the measuring session. All of the trained heifers, regardless of RTH class, successfully accomplished the target training task in 6 sessions, each spending on average 25.3 s per session. All of the trained heifers allowed touches on the rump and perineum at the end of the fourth session. Training nonconfident heifers required more time compared with the others. Trained heifers showed higher RMSSD than nontrained heifers (14.2 vs. 16.9 ms, respectively), indicating a lower vagal tone, and thus, a slightly lower stress level than nontrained heifers. Training did not lead to differences in HR, FCMet, or presence of stress behavioral signs. Nonconfident heifers had the highest mean baseline FCMet values compared with neutral and confident heifers (38.4 vs. 30.3 vs. 29.1 ng/g, respectively). Nonconfident heifers also showed the lowest value of FCMet 12 h after the measuring session (36.7 vs. 44.6 vs. 49.7 ng/g), likely due to a decreased responsiveness of the adrenal gland to a stressor. The average avoidance distance decreased between the beginning and the end of the experiment, especially for neutral and nonconfident heifers, regardless of whether they were trained or not. These results show how using operant conditioning on some heifers not only decreased their vagal tone, but also reduced the responsiveness to humans of all the animals, trained and not trained; in the latter case, this reduction was through nonassociative learning, such as habituation.

A randomized controlled trial assessing the effect of intermittent and abrupt cessation of milking to end lactation on the well-being and intramammary infection risk of dairy cows.

The objectives were to compare the effects of an intermittent milking schedule with a thrice daily milking schedule during the final week of lactation on the well-being, udder health, milk production, and risk of culling of dairy cows. We hypothesized that cows subjected to an intermittent milking schedule would experience less udder engorgement and pain, lower concentrations of fecal glucocorticoid metabolites (11,17-dioxoandrostanes; 11,17-DOA concentration) after dry-off, lower risk of an intramammary infection during the dry period, higher milk production and lower somatic cell count in the subsequent lactation, and lower culling risk compared with herd mates milked 3 times daily and dried off by abrupt cessation. In a randomized controlled field study, Holstein cows (n = 398) with a thrice daily milking schedule were assigned to treatment and control groups. The treatment consisted of an intermittent milking schedule for 7 d before dry-off (gradual cessation of milking, GRAD). Gradual-cessation cows were milked once daily until the day of dry-off, whereas cows in the control group (abrupt cessation of milking, APT) were milked 3 times daily until the day of dry-off. Udder firmness and pain responses of the udder 3 d after dry-off, as well as the percentage change in fecal 11,17-DOA concentration (3 d after dry-off compared with the dry-off day), were used to assess the well-being of the animals. Compared with cows in the GRAD group, the odds [95% confidence interval (CI)] of udder firmness were 1.55 (0.99-2.42) for cows in the APT group, and the odds of a pain response were 1.48 (0.89-2.44) for cows in the APT group. The least squares means (95% CI) of the percentage change in 11,17-DOA concentration were 129.3% (111.1-150.4) for the APT group and 113.6% (97.5-132.4) for the GRAD group. Quarter-level culture results from the periods before dry-off and after calving were compared, to assess the likelihoods of microbiological cure and new infection. Cows in the APT group had lower odds of a new intramammary infection in the dry period [odds ratio, 95% CI: 0.63 (0.37-1.05)], whereas we observed no meaningful differences in the microbiological cure likelihood among groups. The least squares means (95% CI) for somatic cell counts (log10-transformed) were 4.9 (4.8-5.0) in the APT group and 4.9 (4.8-5.0) in the GRAD group. The odds (95% CI) of clinical mastitis in the first 30 d postcalving were 1.32 (0.53-3.30) in the APT group compared with the GRAD group. We observed no meaningful differences in milk production at the first test date postcalving or the culling risk among groups. We conclude that the gradual-cessation protocol tested herein failed to significantly improve animal well-being, udder health, milk production, and survival in the tested study cohort. However, the observed differences in udder firmness, as well as the numerical differences in udder pain and the percentage change in fecal 11,17-DOA concentrations suggest that this line of research may be useful. Future research is needed to develop drying-off strategies that are appropriate for lowering milk production at the end of the lactation and improve animal well-being without compromising udder health.

The effect of transdermal flunixin meglumine on blood cortisol levels in dairy calves after cautery disbudding with local anesthesia.

The objective of this study was to evaluate the effect of the nonsteroidal anti-inflammatory drug transdermal flunixin meglumine (Finadyne Transdermal) on plasma cortisol, average daily weight gain, and standing and lying behavior of calves, when given at the time of disbudding combined with local anesthesia. A sedative was not used to minimize pharmacological interactions. Seventy-one female Holstein Friesian calves aged 13 ± 2 d, with an average weight of 48.9 ± 4.26 kg were enrolled in the study. All calves were randomly assigned to one of 3 treatment groups: (1) control group (CON, n = 27), (2) 1-flunixin group (1-FLU, n = 26) with a single administration of transdermal flunixin meglumine at disbudding, and (3) 2-flunixin group (2-FLU, n = 24) with 2 administrations of transdermal flunixin meglumine, the first treatment at disbudding and the second 6 h after disbudding. Although the CON group received a placebo, 1-FLU and 2-FLU received flunixin meglumine transdermally. To account for plasma cortisol changes due to manipulation and handling of the calves, a sham disbudding procedure was performed one week before disbudding took place. Sham disbudding was conducted by using a cold cautery dehorner applied to each horn bud for 10 s. Disbudding was performed in a similar way by using a hot cautery dehorner. Plasma samples were collected to measure the stress biomarker cortisol at 7 different time points. Body weights were measured 4 times in 2 wk. Standing and lying behavior was assessed via 3-dimensional accelerometer. During sham disbudding and disbudding mean plasma cortisol concentrations were 6.09 ± 2.5 ng/mL and 5.16 ± 2.8 ng/mL, respectively. Treatment tended to have an effect on plasma cortisol concentrations during sham disbudding and had an effect on plasma cortisol concentrations during disbudding. Plasma cortisol concentrations were affected by treatment 2 h after disbudding in comparison to CON group. Furthermore, there was a significant effect on plasma cortisol concentrations 6 h after disbudding in contrast to CON. A return to baseline plasma cortisol levels (initial concentrations) was not achieved in CON during disbudding. There was no statistical difference between average daily weight gain and the treatment procedure. Total lying time was not affected by treatment after disbudding. In conclusion, transdermal flunixin meglumine given at the time of disbudding combined with local anesthesia decreased concentrations of the stress biomarker cortisol, but a second dose 6 h after disbudding had no further effect on plasma cortisol levels.

Endocrine changes related to dog domestication: Comparing urinary cortisol and oxytocin in hand-raised, pack-living dogs and wolves.

Dogs are exceptionally well adapted to life close to humans, and alterations in their endocrine system during the domestication process may be an underlying mechanism. In particular, it has been suggested that low circulating cortisol concentrations in conjunction with simultaneously high oxytocin concentrations may have resulted in dogs' increased docility ('selection for tameness' hypothesis) and heightened propensity to interact and form relationships with humans ('hypersociability' hypothesis) compared to wolves. To investigate this, we analyzed cortisol and oxytocin metabolite concentrations from urine samples of hand-raised, pack-living domestic dogs and their non-domestic relatives, grey wolves. Based on the hypotheses outlined above, we predicted lower cortisol but higher oxytocin concentrations in dogs than wolves. In contrast to our prediction, we found higher cortisol concentrations in dogs than wolves. However, oxytocin concentrations were higher in dogs compared to wolves although the effect was relatively small. Indeed, male dogs had the highest oxytocin concentrations while female dogs' oxytocin concentrations were comparable to wolves'. Feeding status, reproductive phase, and conspecific social interactions also significantly affected cortisol and oxytocin concentrations. Furthermore, we compared two methods of correcting for variable water content of urine samples. We discuss our results in light of physiological and behavioral changes during domestication and highlight the importance of accounting for confounding variables in future studies.

Salivary cortisol responses to acute stress vary between allergic and healthy individuals: the role of plasma oxytocin, emotion regulation strategies, reported stress and anxiety.

Numerous studies have demonstrated that acute psychological stress, induced by the Trier Social Stress Test (TSST) paradigm, affects salivary cortisol secretion and self-reported stress measures including anxiety. Allergy has been related to altered cortisol responsiveness and increased stress vulnerability. Here, we investigated acute stress responses and emotion regulation strategies in cohorts of allergic and healthy individuals. Groups of allergics and healthy individuals were subjected to the TSST and experienced levels of stress and anxiety, as well as emotion regulation strategies, were assessed. Cortisol and oxytocin concentrations were measured in saliva or plasma. The present findings confirm earlier results of altered stress responsiveness in allergic individuals. Acute stress by the TSST evoked higher physiological arousal in allergics by means of salivary cortisol secretion. Allergics also scored higher on emotion suppression. However, individuals who were more likely to use reappraisal recovered more efficiently from the cortisol increase. No such effect for reappraisal was found in the healthy population. No differences in self-reported anxiety and stress emerged between the groups. Plasma oxytocin levels prior to the TSST were significantly higher in allergics. Our data corroborate earlier findings on altered stress susceptibility in allergics. Moreover, we identified differences in emotion regulation and oxytocin secretion which should be further explored. Accounting for the emerging global prevalence of allergy, more in-depth research into the experience of stress, coping strategies and stress-related molecules in allergic people is warranted.Short summaryThis study addressed stress experiences and emotion regulation in allergic and non-allergic adults. Allergics scored higher on emotion suppression, had higher pre-stress concentrations of plasma oxytocin and exhibited a stronger salivary cortisol response to stress than healthy people. The research outcomes indicate that allergic individuals cope less efficiently with acute stress but may benefit from adaptive emotion regulation strategies such as reappraisal.

Stress as a facilitator? Territorial male impala have higher glucocorticoid levels than bachelors.

Territoriality is a common behavioural adaptation, widespread among ungulates. Here, we tested the hypothesis that territorial individuals have higher glucocorticoid concentrations than non-territorial bachelors, in wild impala (Aepyceros melampus) in the Serengeti ecosystem. We also investigated how the relationship between territoriality and glucocorticoid levels is influenced by environmental context, specifically, food quality, population density (i.e., territory defence intensity), and herd size (i.e., mate defence effort). We collected 139 faecal samples over 4 years and analysed these for faecal glucocorticoid metabolites (FGMs). We used Normalised Difference Vegetation Index (NDVI) as a proxy for food quality, and population density was based on aerial surveys. Territorial males had, on average, higher FGM concentrations than bachelors. Increased food quality did not affect FGM levels in territorial males, but decreased FGM levels in bachelors by 78%. Greater population density increased FGM levels by 47%, but this effect was not different between territorial and bachelor males. Herd size did not affect FGM levels in territorial males. While elevated GC levels are often suggested to be repercussions of being territorial, our findings support the hypothesis that elevated GC levels may be beneficial and act as a facilitator of a male's reproductive potential. The elevated GC levels may increase the ability of territorial males to maintain a territory by increasing energy mobilisation and metabolic rate, ultimately increasing their reproductive fitness. Appreciating that long-term increases in GC levels are not simply costly but may have an adaptive, potentially facilitating role in an animal's life history is key to understanding HPA-axis reactivity and its potential in eco-physiological studies.

Herd-level associations between human-animal relationship, management, fecal cortisol metabolites, and udder health of organic dairy cows.

Impact patterns of human-animal relationship (HAR) and herd stress level on udder health were investigated in a cross-sectional study on 30 German and Danish organic dairy herds also taking into account influencing factors regarding housing and management. Cow behavior (avoidance distance, tolerance to tactile interaction, release behavior) was assessed in tests, milkers' behavior recorded during milking, and information about contacts with animals during routine work gathered by interview. Additionally, stockpersons' attitudes were recorded via questionnaires. Fecal cortisol metabolites were measured in approximately 30 focal cows on each farm and used as a proxy to determine the level of distress within the herd. Management and housing were assessed on-farm. The following herd udder health indicators were calculated: the prevalence of mastitis quarters (≥100,000 cells/mL), and, from milk recording data over 1 yr retrospectively, the average somatic cell score and the self-cure rates during lactation per herd. Multivariable regression models with stepwise selection were calculated at herd level. The following HAR-related factors were associated with better udder health (in at least 1 of the final models): stockpersons' higher agreement on patience being important when moving the cows and on necessary contact to cows being pleasant, higher amount of positive interactions with cows during milking, more docile cows in the release behavior test, no routine change of milkers, more contact time during routine work, no active heifer habituation to milking, and performance of barn controls beyond routine work. Lower fecal cortisol metabolite levels were related to higher self-cure rates during lactation. Concerning housing, management, and herd characteristics, the following known factors were related to impaired udder health for at least 1 of the indicators: straw yards, automatic milking system, higher average lactation number, and less antibiotic udder treatments. The results confirm earlier findings that HAR is associated with udder health and should therefore be considered in future research and mastitis control programs. First indications of negative associations between herd stress level and mastitis curing capacity should be followed up in future studies.

Short communication: Pilot study on hormonal, metabolic, and behavioral stress response to treatment of claw horn lesions in acutely lame dairy cows.

Short-term effects of therapeutic claw trimming in acutely lame cows (n=21) with nonadvanced claw horn lesions on the endocrine, metabolic, and behavioral stress responses were investigated in comparison to regular claw trimming in nonlame control cows (n=21). Controls were matched to lame cows by parity and stage of lactation. Lame cows suffering from typical sole ulcers or white line disease were blinded and randomly assigned to 2 treatments, receiving 15 min before interventions either ketoprofen (n=11; 3mg/kg of BW intramuscularly; Romefen, Merial, Lyon, France) or placebo (n=10; saline in equivalent amount and route of administration). All cows underwent functional claw trimming in lateral recumbency on a surgical tipping table, and claw horn lesions in lame cows were conventionally treated (removal of loose horn, block on opposing claw, bandaging of affected claw). Blood samples collected 15 min before, at the end, and 24h after claw trimming were analyzed for concentrations of cortisol, fatty acids, lactate, and glucose, and fecal samples (collected before treatment and after 24 h) for cortisol metabolites. Behavioral stress responses during functional and therapeutic claw trimming were recorded. Concentrations of blood cortisol, fatty acids, glucose, and fecal cortisol metabolites were higher in lame than in nonlame cows after treatment. During claw treatment, more leg movements were recorded for lame cows than nonlame cows. Pre-emptive administration of ketoprofen had no obvious effects on stress responses to therapeutic claw trimming. Treatments of claw horn lesions caused a significant stress and pain reaction in acutely lame cows, demonstrating the necessity of adequate pain management protocols for such interventions.

Welfare and performance in layers following temporary exclusion from the litter area on introduction to the layer facility.

When introduced to the laying facility, pullets are sometimes temporarily excluded from the litter area in order to help them locate food and water, and to prevent floor-laid eggs. This procedure is not permitted in Sweden, because it involves denying access to both litter and space, which may have a negative effect on bird welfare. The present study investigated how the welfare and performance of layers were affected by this temporary exclusion on introduction of hens to the laying facility. The study included 600 floor-reared Dekalb White layers obtained at 16 wk age and housed in 6 groups of 100 in a conventional single-tier floor-laying system. Birds were either given full access to the litter area during the whole study or were excluded from the litter area during the first 2 wk after transfer to the laying facility. From 18 to 72 wk age, birds in both treatments had full access to the litter area. Excluding birds from the litter area for 2 wk resulted in better feather cover and reduced fearfulness, according to novel object and tonic immobility tests. Furthermore, birds initially excluded from the litter area produced eggs with a lower proportion of shell irregularities than birds with full access to the litter area throughout. No difference was found in corticosterone metabolites in droppings rate of lay, mortality, or proportion of floor-laid eggs. In conclusion, none of the parameters studied indicated that the welfare of laying hens was compromised by temporary exclusion from the litter area on introduction to the laying facility. In fact, some of the data suggested that bird welfare had improved.

Age-dependent baseline values of faecal cortisol metabolites in the American mink (Neovison vison) under semi-natural housing conditions.

The welfare of an animal is ensured if it is able to fully satisfy its essential species-typical needs in all functional aspects of behaviour. In mink, stereotypies and apathy, internal and/or external injuries as well as increased susceptibility to disease have been known to occur as a result of chronic stress. The non-invasive method of analysing faecal cortisol metabolites (FCM) allows conclusions to be drawn about the stress level in the respective housing system. The objective of this study is to find out how the cortisol metabolites content in the faecal changes with increasing age of the mink under semi-natural housing conditions. Thus, 40 American mink (Neovison vison) were housed in two outdoor enclosures imitating natural conditions. Throughout the entire study (13th to 32nd week of life), faecal samples were collected to measure cortisol metabolites. No differences in FCM concentrations between the two outdoor enclosures were found. In the young mink lower, less fluctuating FCM levels were found than in older animals. After the first faecal collection in the 13th/14th week of life, the level of metabolites decreased slightly (p = 0.032; 17th/18th week). From the 22nd/23rd week onwards until the 30th/31st week, shortly before the animals were pelted, continuously increasing concentrations were then measured. Increasing FCM levels with advancing age of the animals are probably attributable to the onset of sexual maturity and/or the respective season. This has to be taken into account in future studies using this method for assessing welfare and when comparing different mink housing systems.

Horses' Cardiovascular and Glucocorticoid Responses to Equine-Assisted Therapy with Women with Intellectual Disability: An Exploratory Study.

Research on equine-assisted therapy (EAT) has primarily been centered on human health. Relatively few studies have addressed the impact of EAT on horses. This study sought to monitor four experienced therapy horses' cardiovascular and glucocorticoid activity over the course of standardized EAT sessions designed to support women with intellectual disability. In the control condition, horses completed the EAT protocol solely with the therapist, thereby resembling a training session. Descriptive data analysis revealed higher levels of heart rate during an experimental EAT session and increased salivary cortisol when horses were navigated by the client through an obstacle course during the "challenge" phase of the protocol, pointing at a greater physical demand due to the recipient on horseback. Given the parasympathetic activity and overall heart rate variability across experimental EAT sessions and the cortisol recovery after the sessions, the findings do not give rise to any acute animal welfare concerns. For a more holistic interpretation of the present research results, further investigation into the horse perception of EAT, based on a bigger sample size and additional markers of welfare, is needed.

Introducing young dairy goats into the adult herd after parturition reduces social stress.

The aim of this experiment was to compare social stress, as measured by social behavior and adrenocortical activity, in young dairy goats during the first week after introduction into a herd of adult goats either during the dry period of the herd (i.e., all goats in the herd being pregnant or dry: PD) or shortly after parturition (i.e., all animals lactating or with their kids: LK). Thirty-two young goats that had had no contact with adult goats from the age of 7 wk were introduced into adult goat groups. Adult goats were kept in 2 groups of 36 animals each. Young goats were introduced (in groups of 4 animals each) into each of these 2 groups either during the PD period (2 repetitions) or during LK (2 repetitions); goats with different rearing experience were balanced over introduction periods. Young goats were more often receivers of agonistic social interactions when introduced during PD than during LK. Irrespective of the period of introduction, young goats had other young goats as neighbors more frequently than expected by chance alone, although this was even more distinct during PD. Cortisol metabolite levels increased markedly from baseline during PD, but not after parturition. Rearing showed an effect only on the nearest neighbors, with mother-reared young goats staying closer together. Our results indicate that young goats experience less social stress when being introduced into a herd of adult dairy goats shortly after parturition and with kids still present rather than during the dry period. Whether this effect is due to the period and lactational stage itself or to the presence of kids needs to be tested in future studies.

Long-term effects of early maternal deprivation on goat social behaviour.

Early maternal deprivation has been shown to disrupt goat kids' social behaviour and stress-coping strategy, and has long-term effects in other species like cattle. We studied the long-term effects of early maternal deprivation on 18-month-old goats. Seventeen goats were raised together with their dams (DR kids) and other lactating goats and kids, while 18 goats were separated from their dams three days after birth and artificially reared together (AR kids). Kids of both treatments were weaned around two to three months of age, grouped and raised together until this study 15 months later. Affiliative, playful, and agonistic behaviour was recorded by focal sampling in the home pen, when the focal goat had rejoined the herd after being physically isolated for 3 minutes, and after the focal goat was restrained and manipulated for 3 minutes. Behavioural observations were also conducted after the goats were introduced in groups of four in a herd of 77 unknown, lactating multiparous goats. Avoidance distance tests were performed in the home pen to assess the human-animal relationship. Salivary cortisol was measured before and after physical isolation, and faecal glucocorticoid metabolites were measured before and 24 hours after introduction in the lactating herd. In the home pen, AR goats were involved in less head nudging than DR goats, but other social behaviours or their behavioural and physiological response to the various stressful situations were not affected by their rearing treatment. Upon introduction in a DR lactating herd, most of the agonistic interactions observed were initiated by multiparous goats towards the introduced AR and DR goats alike. AR goats received more threats from the multiparous goats than DR goats, but were involved in less clashing than DR goats. AR goats showed less avoidance of familiar and unfamiliar humans than DR goats. Overall, previously AR and DR goats showed only a few differences in affiliative and agonistic behaviours in their home pen or after being exposed to different stressors 15 months later. Nonetheless, after being introduced into a multiparous goat herd, AR goats were still threatened more often than DR goats, and DR goats clashed more than AR goats, suggesting some persistent differences in their social abilities observed at an early age before and after weaning. As predicted, AR goats remained less fearful of humans than DR goats.

Antidepressants recruit new neurons to improve stress response regulation.

Recent research suggests an involvement of hippocampal neurogenesis in behavioral effects of antidepressants. However, the precise mechanisms through which newborn granule neurons might influence the antidepressant response remain elusive. Here, we demonstrate that unpredictable chronic mild stress in mice not only reduces hippocampal neurogenesis, but also dampens the relationship between hippocampus and the main stress hormone system, the hypothalamo-pituitary-adrenal (HPA) axis. Moreover, this relationship is restored by treatment with the antidepressant fluoxetine, in a neurogenesis-dependent manner. Specifically, chronic stress severely impairs HPA axis activity, the ability of hippocampus to modulate downstream brain areas involved in the stress response, the sensitivity of the hippocampal granule cell network to novelty/glucocorticoid effects and the hippocampus-dependent negative feedback of the HPA axis. Remarkably, we revealed that, although ablation of hippocampal neurogenesis alone does not impair HPA axis activity, the ability of fluoxetine to restore hippocampal regulation of the HPA axis under chronic stress conditions, occurs only in the presence of an intact neurogenic niche. These findings provide a mechanistic framework for understanding how adult-generated new neurons influence the response to antidepressants. We suggest that newly generated neurons may facilitate stress integration and that, during chronic stress or depression, enhancing neurogenesis enables a dysfunctional hippocampus to restore the central control on stress response systems, then allowing recovery.

Circadian activity of the hypothalamic-pituitary-adrenal axis is differentially affected in the rat chronic mild stress model of depression.

The altered activity of the hypothalamic-pituitary-adrenal (HPA) axis is often observed in stress-related disorders. According to the literature, about 60% of patients with major depressive disorder elicit high levels of cortisol. It is still unclear why high cortisol levels are not observed in all patients. In this study, we used the chronic mild stress (CMS) rat model of depression, which is based on continuous exposure to unpredictable stressors, to track longitudinal changes in HPA function using fecal corticosterone metabolites (FCM) as a read out. The dexamethasone suppression test was used to assess negative feedback inhibition of the HPA axis. Our results show (1) a disturbance in diurnal corticosterone rhythm measured as fluctuations of the diurnal FCM peak, (2) differences in corticosterone levels between stress-susceptible and stress-resilient animals, (3) recovery of diurnal corticosterone rhythm after 8 weeks of CMS, and (4) alterations in sensitivity to dexamethasone in negative feedback regulation of corticosterone secretion during the time course of CMS. Thus, a disruption of HPA axis circadian rhythmicity coincides with the initial state in the development of depression-like behavior. This chronobiological abnormality, as well as the hypersecretion of corticosterone, is state, rather than trait, dependent.

Effects of separated pair housing of female C57BL/6JRj mice on well-being.

In laboratory animal facilities, it is a common code of practice to house female mice in groups. However, some experimental conditions require to house them individually, even though social isolation may impair their well-being. Therefore, we introduced a separated pair housing system and investigated whether it can refine single housing of adult female C57BL/6JRj mice. Individually ventilated cages (IVC) were divided by perforated transparent walls to separate two mice within a cage. The cage divider allowed visual, acoustic, and olfactory contact between the mice but prevented interindividual body-contact or food sharing. Short- and long-term effects of the separated pair housing system on the well-being of the mice were compared with single and group housing using a range of behavioral and physiological parameters: Nest building behavior was assessed based on the complexity of nests, the burrowing performance was measured by the amount of food pellets removed from a bottle, and trait anxiety-related behavior was tested in the free exploratory paradigm. For the evaluation of the ease of handling, interaction with the experimenter's hand was monitored. Social interaction with unknown conspecifics and locomotor activity were investigated in a test arena. Moreover, body weight and stress hormone (metabolites) were measured in feces and hair. After the mice spent a day under the respective housing conditions, concentrations of fecal corticosterone metabolites were higher in separated pair-housed mice, and they built nests of a higher complexity when compared to single-housed mice. The latter effect was still observable eight weeks later. In week 8, separated pair-housed mice showed less locomotor activity in the social interaction arena compared to mice from the other housing systems, i.e., single and group housing. Regardless of the time of testing, pair housing improved the burrowing performance. Separated pair-housed mice were more difficult to catch than group-housed mice. Hair corticosterone, progesterone, and dehydroepiandrosterone concentrations changed with increasing age independently of the housing system. There were no effects of the housing systems on trait anxiety-related behavior in the free exploratory paradigm, voluntary interaction with the experimenter's hand, and body weight. Overall, the transfer to the separated pair housing system caused short-term stress responses in female C57BL/6JRj mice. Long-term effects of separated pair housing were ambiguous. On one hand, separated pair housing increased nesting and burrowing behavior and may therefore be beneficial compared to single housing. But on the other hand, locomotor activity decreased. The study underlined that the effects of the housing conditions on physiological and behavioral parameters should be considered when analyzing and reporting animal experiments.

[Heart rate and faecal cortisol metabolites measurements in horses at the Sechseläuten in Zurich]. / Messung von Herzfrequenzen und Kotcortisolmetaboliten bei Pferden am Zürcher Sechseläuten.

INTRODUCTION: The Zurich's Sechseläuten is a traditional festival of the Zurich guilds, in which around 500 horses take part. After a parade through the old town of Zurich, the riders gather at a big square to canter around a burning woodpile topped with an exploding effigy (the «Böögg¼). The level of stress experienced by the horses partaking in this event is subjected to increasing scrutiny. The aim of this study was to evaluate the stress load of the horses participating in the Sechseläuten procession more objectively by measuring heart rate and faecal cortisol metabolites. Twenty-three horse-rider pairs were voluntarily recruited from the guilds for participation in the study. For comparison purposes, three sections of the procession were simulated in terms of gait and distance travelled in a normal riding environment during a test day and observed during the Sechseläuten itself 16 days later: Counter march Bahnhofstrasse, parade Limmatquai and parade around the burning «Böögg¼. All horses were equipped with a heart rate measuring system with GPS tracking (Polar Team Pro®, Polar Electro Oy, Finland). Faecal samples for measuring cortisol metabolites were taken rectally immediately before and 24 hours after the events. Thirteen of the 23 horses were sedated during the Sechseläuten (Acepromazine, 0,1-0,2 mg/kg p.o.). The differences between the observation periods and groups (sedated/unsedated) were tested by analysis of variance (P.

INTRODUCTION: Le Sechseläuten de Zurich est une fête traditionnelle des guildes zurichoises à laquelle participent environ 500 chevaux. Après un défilé dans la vieille ville de Zurich, les cavaliers se rassemblent sur une grande place pour galoper autour d'un tas de bois en feu surmonté d'une effigie qui explose (le «Böögg¼). Le niveau de stress des chevaux participant à cet événement fait l'objet d'une attention croissante. L'objectif de cette étude était d'évaluer plus objectivement le niveau de stress des chevaux participant au cortège du Sechseläuten en mesurant la fréquence cardiaque et les métabolites fécaux du cortisol. Vingt-trois couples cheval-cavalier ont été recrutés volontairement dans les guildes pour participer à l'étude. À des fins de comparaison, trois sections du cortège ont été simulées en termes d'allure et de distance parcourue dans un environnement équestre normal lors d'une journée de test et observées ensuite pendant le Sechseläuten lui-même 16 jours plus tard: Contre-marche sur la Bahnhofstrasse, cortège sur le Limmatquai et chevauchée autour du «Böögg¼ en feu. Tous les chevaux ont été équipés d'un système de mesure de la fréquence un cardiomètre avec suivi GPS (Polar Team Pro®, Polar Electro Oy, Finlande). Des échantillons fécaux destinés à mesurer les métabolites du cortisol ont été prélevés par voie rectale immédiatement avant et 24 heures après les événements. Treize des 23 chevaux ont été mis sous sédation pendant le Sechseläuten (Acépromazine, 0,1­0,2 mg/kg p.o.). Les différences entre les périodes d'observation et les groupes (sédaté/non-sédaté) ont été testées par analyse de variance (P.

Heifers don't care: no evidence of negative impact on animal welfare of growing heifers when using virtual fences compared to physical fences for grazing.

Virtual fencing (VF) represents a way to simplify traditional pasture management with its high labour and cost requirements for fencing and to make better use of the 'beneficial' agronomic and ecological effects of livestock grazing. In this study, the VF technology (® Nofence, AS, Batnfjordsøra Norway) was used with Fleckvieh heifers to investigate possible welfare impacts on the animals compared to conventionally fenced animals when they were trained to respond correctly to the system. The Nofence® collars (attached to the neck of the heifers) send acoustic signals as a warning when the animals approach the VF line, which was set up by GPS coordinates within the Nofence®-App, followed by an electric pulse when they do not stop or return. The heifers had no experience with VF prior to the study. Two treatments (VF versus physical fencing (PF)) were applied to six groups of four heifers each (three groups per treatment) over three 12-day time replicates. One VF line separated the pasture of the VF group into an accessible or non-accessible area. The control group had a PF line. Both groups were equipped with Nofence® collars (deactivated for the PF group). The trial took place on two adjacent paddocks of 1 000 m2 each following a 12-day schedule which was divided into three sections: visual support of the VF line by a physical barrier (first 2 days), only virtual border without visual support, moving the VF line (on day 8). Each time replicate followed the next successively on different paddocks with two new groups of heifers, which were grazed 5 h daily. During the whole experiment, the behaviour of each of the four animals per group was continuously observed; 2 h a.m., 2 h p.m. Exclusion by the VF line was effective in our trial. None of the heifers crossed the virtual boundary, i.e. the time spent in exclusion zone was zero. The heifers received 2.70 ± 2.63 acoustic signals and 0.30 ± 0.36 electric pulses (mean ± SD) per heifer and hour during all time replicates. Main cattle behaviour on pasture was not affected by the fencing system. Live weight gain, herbage consumption and faecal cortisol metabolites also revealed no significant differences. The duration until the heifers restarted grazing after an electric pulse from the Nofence® collar was significantly shorter than after an electric pulse from the physical fence. We can summarise that in our study, cattle well-being on pasture was not negatively affected by VF compared to PF.

Stress and stereotypic behaviour in mink (Mustela vison): a focus on adrenocortical activity.

We examined whether female mink with low (LS) and high (HS) occurrence of stereotypic behaviour differ in their adrenocortical activity in baseline conditions or in response to immobilisation (Experiment 1), handling, adrenocorticotropic hormone (ACTH) challenge (Experiment 2) and excretion of circulating cortisol (Experiment 3). Faeces are the predominating excretory route of cortisol (83%), with peak concentrations after 4.2 h (urine: 3.4 h). Faecal cortisol metabolites (FCMs) reflected changes in relation to handling/ACTH challenge. In Experiment 1 (n = 162), HS mink had approximately 54% higher baseline level of FCM than LS mink (P < 0.001), with markedly elevated FCM on the days after an immobilisation stressor. In Experiment 2 (n = 48), LS and HS mink did not differ in adrenocortical activity after an ACTH challenge. However, HS mink reacted more in response to handling, evident in the FCM level 4-20 h after the handling (P = 0.001). In Experiment 3 (n = 16), the excretion of infused (3)H-cortisol did not differ between LS and HS mink. Stereotypic behaviour is concurrent with higher baseline concentrations of FCM, which cannot be explained by a greater adrenocortical reactivity or a different excretion of the circulating cortisol. Instead, we conclude that mink with a high level of stereotypic behaviour have a greater perception of stress, or increased sensitivity to stressors at the pituitary level.