A review of pain assessment techniques and pharmacological approaches to pain relief after bovine castration: Practical implications for cattle production within the United States

Castration of male calves destined for beef production is a common livestock management practice in the United States amounting to approximately 7 million procedures per year. Recently there has been renewed interest in identifying methods to reduce pain associated with dehorning and castration. Although several studies have reported that analgesic drug administration prior to castration attenuates plasma cortisol response, there are currently no compounds specifically approved for pain relief in livestock in the U.S. Validated pain assessment tools are needed to support regulatory approval of analgesic compounds. This may include use of accelerometers, videography, heart rate variability determination, electroencephalography, thermography and plasma neuropeptide measurement to assess behavioral, physiological and neuroendocrine changes associated with a pain response. Extra-label drug use (ELDU) for pain relief is regulated under the Animal Medicinal Drug Use Clarification Act (AMDUCA) and requires that drugs be administered by or under the supervision of a veterinarian. Agents that may provide preemptive analgesia include local anesthetics, nonsteroidal anti-inflammatory drugs (NSAIDs), opioids, α2-agonists, and N-methyl d-aspartate (NMDA) receptor antagonists. A review of the published literature suggests that a significant decrease in plasma cortisol concentration after castration was associated with preemptive administration of a NSAID and local anesthesia. Local anesthesia alone tended to decrease peak plasma cortisol concentrations more than NSAIDs. However NSAIDs alone tended to decrease the area under the plasma cortisol-time curve more than local anesthesia alone. These findings suggest that multimodal analgesic regimens that extend into the post-operative period are more effective at mitigating pain and distress associated with castration than a single drug modality. Regulatory approval of safe and cost effective analgesic compounds with convenient routes of administration is needed for routine use of pain relieving drugs to be considered as standard practice at the time of castration.     

Effect of rectal or intravenous tramadol on the incidence of pain-related behaviour after disbudding calves with caustic paste

To evaluate the efficacy of tramadol (suppositories or intravenous) in alleviating pain after caustic paste disbudding, two behaviour-assessing methods were used during the first hour post-disbudding: the objective recording of four pain-related behaviours and the subjective scoring pain on a scale of 0-10 (Numerical Rating Scale, NRS). Sixty dairy calves were allocated to five groups: disbudded with no treatment (C); disbudded after rectal tramadol (TR-r); disbudded after i.v. tramadol (TR-iv); sham-disbudded after i.v. tramadol (TR-SD); sham-disbudded with no treatment (SD). All disbudded groups showed more head shakes, head rubs and transitions for the first 15 min when compared with sham-disbudded calves. Animals treated with rectal tramadol showed more head shakes, head rubs and transitions than all other groups from 15 to 30 min after disbudding. Ear flicks were more frequent in disbudded calves than in sham-disbudded ones at the end of the observation period (45-60 min). NRS showed a higher score at all moments for all disbudded groups when compared with sham-disbudded. Sham-disbudded animals with i.v. tramadol had a higher score than sham-disbudded with no treatment at the first observation period (15 min). We concluded that caustic paste disbudding causes severe pain for the first 30 min after applying the paste and that tramadol is not efficient in controlling this pain but when given i.v. it may help in reducing post-operative nociception. We also showed that tramadol can be used safely in young calves although after i.v. administration it may cause some sham-disbudded animals to shake their heads and ears during the first minutes after injection. NRS showed to be a reliable method of evaluating the acute pain in caustic paste disbudded calves.     

Pharmacokinetics and effect of intravenous meloxicam in weaned Holstein calves following scoop dehorning without local anesthesia

ABSTRACT: BACKGROUND: Dehorning is a common practice involving calves on dairy operations in the United States. However, less than 20% of producers report using analgesics or anesthetics during dehorning. Administration of a systemic analgesic drug at the time of dehorning may be attractive to dairy producers since cornual nerve blocks require 10 -- 15 min to take effect and only provide pain relief for a few hours. The primary objectives of this trial were to (1) describe the compartmental pharmacokinetics of meloxicam in calves after IV administration at 0.5 mg/kg and (2) to determine the effect of meloxicam (n = 6) or placebo (n = 6) treatment on serum cortisol response, plasma substance P (SP) concentrations, heart rate (HR), activity and weight gain in calves after scoop dehorning and thermocautery without local anesthesia. RESULTS: Plasma meloxicam concentrations were detectable for 50 h post-administration and fit a 2-compartment model with a rapid distribution phase (mean T1/2alpha = 0.22 +/- 0.087 h) and a slower elimination phase (mean T1/2beta = 21.86 +/- 3.03 h). Dehorning caused a significant increase in serum cortisol concentrations and HR (P < 0.05). HR was significantly lower in the meloxicam-treated calves compared with placebo-treated calves at 8 h (P = 0.039) and 10 h (P = 0.044) after dehorning. Mean plasma SP concentrations were lower in meloxicam treated calves (71.36 +/- 20.84 pg/mL) compared with control calves (114.70 +/- 20.84 pg/mL) (P = 0.038). Furthermore, the change in plasma SP from baseline was inversely proportional to corresponding plasma meloxicam concentrations (P = 0.008). The effect of dehorning on laying behavior was less significant in meloxicam-treated calves (p = 0.40) compared to the placebo-treated calves (P < 0.01). Calves receiving meloxicam prior to dehorning gained on average 1.05 +/- 0.13 kg bodyweight/day over 10 days post-dehorning compared with 0.40 +/- 0.25 kg bodyweight/day in the placebo-treated calves (p = 0.042). CONCLUSIONS: To our knowledge, this is the first published report examining the effects of meloxicam without local anesthesia on SP, activity and performance of calves post-dehorning. These findings suggest that administration of meloxicam alone immediately prior to dehorning does not mitigate signs of acute distress but may have long term physiological, behavior and performance effects.     

Effect of previous handling experiences on responses of dairy calves to routine husbandry procedures

The nature of human–animal interactions is an important factor contributing to animal welfare and productivity. Reducing stress during routine husbandry procedures is likely to improve animal welfare. We examined how the type of early handling of calves affected responses to two common husbandry procedures, ear-tagging and disbudding. Forty Holstein–Friesian calves (n = 20/treatment) were exposed to one of two handling treatments daily from 1 to 5 weeks of age: (1) positive (n = 20), involving gentle handling (soft voices, slow movements, patting), and (2) negative (n = 20), involving rough handling (rough voices, rapid movements, pushing). Heart rate (HR), respiration rate (RR) and behaviour (activity, tail flicking) were measured before and after ear-tagging and disbudding (2 days apart). Cortisol was measured at −20 (baseline), 20 and 40 min relative to disbudding time. There were no significant treatment differences in HR, RR or behaviour in response to either procedure. However, the following changes occurred across both treatment groups. HR increased after disbudding (by 14.7 ± 4.0 and 18.6 ± 3.8 bpm, positive and negative, respectively; mean ± s.e.m.) and ear-tagging (by 8.7 ± 3.1 and 10.3 ± 3.0 bpm, positive and negative, respectively). After disbudding, there was an increase in RR (by 8.2 ± 3.4 and 9.3 ± 3.4 breaths/min, positive and negative, respectively), overall activity (by 9.4 ± 1.2 and 9.9 ± 1.3 frequency/min, positive and negative, respectively) and tail flicking (by 13.2 ± 2.8 and 11.2 ± 3.0 frequency/min, positive and negative, respectively), and cortisol increased from baseline at 20 min post procedure (by 10.3 ± 1.1 and 12.3 ± 1.1 nmol/l positive and negative, respectively). Although we recorded significant changes in calf responses during ear-tagging and disbudding, the type of prior handling had no effect on responses. The effects of handling may have been overridden by the degree of pain and/or stress associated with the procedures. Further research is warranted to understand the welfare impact and interaction between previous handling and responses to husbandry procedures.     

Effects of a local anaesthetic and NSAID in castration of piglets,on the acute pain responses,growth and mortality

The present study addresses the questions whether on-farm use of local anaesthesia with lidocaine leads to a reduction in pain responses during castration, and whether the non-steroidal anti-inflammatory drug meloxicam improves technical performance after castration of piglets. Five treatments were included in the study: (1) castration without anaesthesia or analgesia (CAST), (2) castration after local anaesthesia with lidocaine (LIDO), (3) castration after administration of meloxicam (MELO), (4) castration after lidocaine and meloxicam (L + M) and (5) sham castration (SHAM). To reduce litter influences, each treatment was present in each of the 32 litters (n = 32 per treatment). During castration, vocalizations were recorded continuously. Blood samples were collected 15 min before and 20 min after castration for determination of plasma levels of total cortisol, glucose, lactate and creatine kinase (CK). Mortality was registered and piglets were weighed several times to calculate growth. Several aspects of vocalizations during castration showed consistent and significantly different levels in CAST compared with LIDO, L + M and SHAM. CAST piglets squealed longer, louder and higher. Vocalizations of MELO piglets most resembled those of CAST. An increase in cortisol was seen in all treatments. However, in SHAM piglets this increase was significantly lower than in the other treatments. LIDO piglets showed a significantly smaller increase in plasma cortisol levels compared with CAST and MELO. L + M piglets differed significantly only from the SHAM group. Lactate levels differed significantly between LIDO and MELO, the level in LIDO being decreased after castration. In the other treatments an increase was measured. No treatment effects were found in plasma glucose and CK levels, nor in growth and mortality of the piglets. In conclusion, on the basis of vocalizations and plasma cortisol, local anaesthesia with lidocaine reduces pain responses in piglets during castration. A positive effect of meloxicam on technical performance was not found.     

Minimising pain in farm animals: the 3S approach – ‘Suppress,Substitute, Soothe’

Recently, the French National Institute for Agricultural Research appointed an expert committee to review the issue of pain in food-producing farm animals. To minimise pain, the authors developed a ‘3S’ approach accounting for ‘Suppress, Substitute and Soothe’ by analogy with the ‘3Rs’ approach of ‘Reduction, Refinement and Replacement’ applied in the context of animal experimentation. Thus, when addressing the matter of pain, the following steps and solutions could be assessed, in the light of their feasibility (technical constraints, logistics and regulations), acceptability (societal and financial aspects) and availability. The first solution is to suppress any source of pain that brings no obvious advantage to the animals or the producers, as well as sources of pain for which potential benefits are largely exceeded by the negative effects. For instance, tail docking of cattle has recently been eliminated. Genetic selection on the basis of resistance criteria (as e.g. for lameness in cattle and poultry) or reduction of undesirable traits (e.g. boar taint in pigs) may also reduce painful conditions or procedures. The second solution is to substitute a technique causing pain by another less-painful method. For example, if dehorning cattle is unavoidable, it is preferable to perform it at a very young age, cauterising the horn bud. Animal management and constraint systems should be designed to reduce the risk for injury and bruising. Lastly, in situations where pain is known to be present, because of animal management procedures such as dehorning or castration, or because of pathology, for example lameness, systemic or local pharmacological treatments should be used to soothe pain. These treatments should take into account the duration of pain, which, in the case of some management procedures or diseases, may persist for longer periods. The administration of pain medication may require the intervention of veterinarians, but exemptions exist where breeders are allowed to use local anaesthesia (e.g. castration and dehorning in Switzerland). Extension of such exemptions, national or European legislation on pain management, or the introduction of animal welfare codes by retailers into their meat products may help further developments. In addition, veterinarians and farmers should be given the necessary tools and information to take into account animal pain in their management decisions.     

Benefits of a multimodal analgesia compared to local anesthesia alone to alleviate pain following castration in sheep: a multiparametric approach

Castration of male ruminants is a common livestock management practice, but induces pain. However, little is known about the effectiveness of multimodal analgesia compared to local anesthesia (LA) alone in reducing pain associated with burdizzo castration in sheep. This study aimed to monitor the pain response induced by castration in sheep and to assess the efficacy of analgesia strategies. Twenty-four 12-month-old male Texel sheep were burdizzo-castrated after administration of physiological serum (Burd), local anesthetic (Burd+LA) or LA plus non-steroidal anti-inflammatory drug (Burd+LA+NSAID). Sheep responses were monitored using behavioral and physiological indices of pain. Sampling occurred from 24 h pre-castration to 78 h post-castration, split into four periods based on the duration of analgesia: P0 (T-24 to T-1 h), P1 (T0 to T+2 h), P2 (T+3 to T+32 h) and P3 (T+36 to T+78 h). Behavioral indices were attention and head position, ear position, position of the eyelid, other facial expression, standing/lying postures, postures of the legs, clinical signs and abnormal activities. Physiological indices consisted in indicators of inflammation (haptoglobin, serum amyloid A (SAA), body temperature), hypothalamo–pituitary–adrenal axis (cortisol, non-esterified fatty acids, glucose), autonomous nervous system (heart rate variability (HRV)) and oxidative stress. The variables contributing most to discrimination of the period×treatment groups were analyzed by factorial discrimination analysis. Pre-castration (P0), there was no significant difference between treatments for all indicators (P > 0.05). Post-castration, eight indicators varied significantly according to period and treatment: cortisol, clinical signs, ratio of low frequency/high frequency (LF/HF) bands of the HRV, attention and head position, SAA, haptoglobin, body temperature and glucose. The treatment×periods groups were well discriminated by the 23 indicators. Burd in P0, Burd+LA in P0 and Burd+LA+NSAID in P0, P1 and P2 had low values for all indicators, likely reflecting absence of pain and discomfort. Burd in P1 and P2 and Burd+LA in P2 showed clinical signs and reduced attention, high LF/HF and high cortisol levels, reflecting acute pain. Burd and Burd+LA in P3 had high temperature, high haptoglobin, high glucose and high SAA, but no response from other pathways. These results suggest that (i) behavioral signs of pain were apparent up to 32 h post-castration, (ii) LA was partially effective, but only during its time of action (2 h) and (iii). multimodal analgesia (LA and NSAID) was effective for up to 3 days post-castration. These findings, and especially those related to sheep behavior, can help veterinarians and farmers better detect pain and refine their pain alleviation methods.     

Physiological and genetic correlates of boldness: characterising the mechanisms of behavioural variation in rainbow trout, Oncorhynchus mykiss

Bold, risk-taking animals have previously been putatively linked with a proactive stress coping style whereas it is suggested shyer, risk-averse animals exhibit a reactive coping style. The aim of this study was to investigate whether differences in the expression of bold-type behaviour were evident within and between two lines of rainbow trout, Oncorhynchus mykiss, selectively bred for a low (LR) or high (HR) endocrine response to stress, and to link boldness and stress responsiveness with the expression of related candidate genes. Boldness was determined in individual fish over two trials by measuring the latency to approach a novel object. Differences in plasma cortisol concentrations and the expression of eight novel candidate genes previously identified as being linked with divergent behaviours or stress were determined. Bold and shy individuals, approaching the object within 180 s or not approaching within 300 s respectively, were evident within each line, and this was linked with activity levels in the HR line. Post-stress plasma cortisol concentrations were significantly greater in the HR line compared with the LR line, and six of the eight tested genes were upregulated in the brains of LR fish compared with HR fish. However, no direct relationship between boldness and either stress responsiveness or gene expression was found, although clear differences in stress physiology and, for the first time, gene expression could be identified between the lines. This lack of correlation between physiological and molecular responses and behavioural variation within both lines highlights the complexity of the behavioural-physiological complex.     

Sedation or anaesthesia decrease the stress response to electroejaculation and improve the quality of the collected semen in goat bucks

Electroejaculation (EE) is stressful and probably painful; thus the administration of anaesthesia is recommended to decrease those negative effects. However, anaesthesia has a direct risk of provoking death, but sedation is less risky than anaesthesia. At the same time, α2-adrenergic agonists may improve semen quality. Therefore, the aim of the present study was to compare the physiological and behavioural responses indicative of stress and possibly pain, and the semen quality in electroejaculated untreated, anaesthetised or sedated goat bucks. Semen was collected from eight bucks using three different procedures in all them (EE in untreated bucks, EE under sedation or EE under general anaesthesia). The number of vocalizations during EE and the behavioural pattern before and after procedures were recorded. Pain visual analogue scale (VAS) score was also determined during EE. Rectal temperature, heart rate, serum cortisol concentration, biochemical and haematological parameters were measured before and after each procedure, and sperm characteristics were determined. Bucks vocalised more often when untreated than sedated or anaesthetised (P<0.02). The pain VAS score was greater when bucks were untreated than sedated or anaesthetised (P<0.002). The rectal temperature, heart rate, total protein, albumin and haemoglobin concentrations were greater when bucks were untreated than anaesthetised or sedated (P<0.02). Serum cortisol increased after EE (P=0.0006), without differences between procedures. The frequency and duration of lying down after EE were greater when bucks were anaesthetised than sedated or untreated (P<0.05), and were also greater when bucks were sedated than untreated (P<0.05). The number of times that the animal tried to stand up after EE was greater when bucks were anaesthetised than sedated or untreated (P<0.02). The sperm mass motility was greater when bucks were anaesthetised or sedated than when they were untreated (P=0.048). When animals were sedated, the ejaculate contained more spermatozoa with functional plasma membrane (P=0.03) and morphologically normal (P=0.05) than when they were untreated. In conclusion, general anaesthesia and sedation decreased the stress and probably the pain response provoked by EE and especially sedation improved the quality of the semen collected.     

Effects of rapid experimental temperature increases on acute physiological stress and behaviour of stream dwelling juvenile chinook salmon

We experimentally heated small streams in summer and investigated the short-term behavioural changes and physiological stress responses of juvenile chinook salmon (Oncorhynchus tshawytscha). We rapidly raised temperatures ∼1–4 °C for 1.5 h above ambient levels of ∼7–15 °C in groundwater fed tributary streams and ∼19–23 °C in side-channel streams. Juvenile chinook rearing in groundwater fed tributaries were generally unaffected behaviourally; however, we found that temperature increase caused fish in the tributary trials to be physiologically stressed (elevations in mean cortisol concentrations ranged from 116% to 253%). Side-channel trials caused some mortality of juvenile chinook and a stronger display of behaviours indicative of stress and avoidance such as erratic swimming, abnormal posture, and aggregative behaviour. Foraging rates increased over 56 times in response to heating in side-channel trials. Cortisol levels did not increase in side-channel trials, but rather showed a trend to levels below control values suggesting an impaired stress response possibly due to chronic stress. Our results may reflect conservative responses in terms of what we may find with other salmonid species since juvenile chinook have been described as the most tolerant of the Pacific salmon species to elevated temperatures.     

A multiparametric approach to discriminate the impacts of different degrees of invasiveness of surgical procedures in sheep

Traumatic situations in animals induce responses including pain, expressed through behavioural and physiological pathways such as inflammation, oxidative stress, hypothalamic–pituitary–adrenal (HPA) axis and autonomic nervous system. As some of these systems can also be activated during excitement and situations with a positive valence, their use as a means to assess pain response is difficult. We explored (i) how these five aforementioned pathways change in sheep exposed to various degrees of invasiveness of surgical procedures despite a therapeutic regimen and (ii) whether a multiparametric analysis that combines information from these five pathways enhances the discrimination between these situations, and estimates the relative importance of these pathways in the response. We used 24 adult sheep split into four treatments: Control (C; no fasting, no anaesthesia, no surgery), Sham (S; fasting, anaesthesia, no surgery), Rumen Canulation (R; fasting, anaesthesia, rumen cannulation) and Rumen–Duodenal–Ileum cannulation (RDI; fasting, anaesthesia, cannulation of the rumen, duodenum and ileum). Sheep’ responses were measured for 5 days after surgery. When considering each behavioural or physiological pathway independently, discrimination between treatments was acceptable, its sensitivity (Se) ranging from 0% to 100%, and its specificity (Sp) ranging from 62% to 100%. The multiparametric analysis gathering information from the five pathways enhanced the effectiveness of discrimination between treatments (Se, 50% to 100%; Sp, 82% to 100%), and gave additional information on the relative contribution of each pathway to the global sheep response. Sheep global response was higher when exposed to a surgery, and increased with the surgery invasiveness. This response relied mostly on inflammation (absolute correlation for haptoglobin, 0.89), HPA (cortisol, 0.85) and behaviour (antalgic postures, 0.85). The multiparametric approach seems to be a promising tool to discriminate between different degrees of invasiveness of surgical procedures.     

Stress effects on AVT and CRF systems in two strains of rainbow trout (Oncorhynchus mykiss) divergent in stress responsiveness

The aim for this study was to examine whether the F4 generation of two strains of rainbow trout divergent in their plasma cortisol response to confinement stress (HR: high responder or LR: low responder) would also differ in stress-induced effects on forebrain concentrations of mRNA for corticotropin-releasing factor (CRF), arginine vasotocin (AVT), CRF receptor type 1 (CRF-R1), CRF receptor type 2 (CRF-R2) and AVT receptor (AVT-R). In addition, plasma cortisol concentrations, brainstem levels of monoamines and monoamine metabolites, and behaviour during confinement were monitored. The results confirm that HR and LR trout differ in their cortisol response to confinement and show that fish of these strains also differ in their behavioural response to confinement. The HR trout displayed significantly higher locomotor activity while in confinement than LR trout. Moreover, following 180 min of confinement HR fish showed significantly higher forebrain concentrations of CRF mRNA than LR fish. Also, when subjected to 30 min of confinement HR fish showed significantly lower CRF-R2 mRNA concentrations than LR fish, whereas there were no differences in CRF-R1, AVT or AVT-R mRNA expression between LR and HR fish either at 30 or 180 min of confinement. Differences in the expression of CRF and CRF-R2 mRNA may be related to the divergence in stress coping displayed by these rainbow trout strains.     

Pain and Pessimism: Dairy Calves Exhibit Negative Judgement Bias following Hot-Iron Disbudding

Pain is defined as an unpleasant sensory and emotional experience associated with actual or potential tissue damage, but emotional states are difficult to directly assess in animals. Researchers have assessed pain using behavioural and physiological measures, but these approaches are limited to understanding the arousal rather than valence of the emotional experience. Cognitive bias tasks show that depressed humans judge ambiguous events negatively and this technique has been applied to assess emotional states in animals. However, limited research has examined how pain states affect cognitive processes in animals. Here we present the first evidence of cognitive bias in response to pain in any non-human species. In two experiments, dairy calves (n = 17) were trained to respond differentially to red and white video screens and then tested with unreinforced ambiguous colours in two or three test sessions before and two sessions after the routine practice of hot-iron disbudding. After disbudding calves were more likely to judge ambiguous colours as negative. This ‘pessimistic’ bias indicates that post-operative pain following hot-iron disbudding results in a negative change in emotional state.     

Measurement of cortisol,cortisone, prednisolone,dexamethasone and 11-deoxycortisol with ultra high performance liquid chromatography–tandem mass spectrometry: Application for plasma,plasma ultrafiltrate,urine and saliva in a routine laboratory

We describe an ultra high performance liquid chromatography–tandem mass spectrometry (UHPLC MS/MS) method suitable for a routine laboratory to determine endogenous and exogenous glucocorticoids in plasma, plasma ultrafiltrate, urine and saliva in a single analytical run. After addition of a multi-analyte internal standard, a standardised sample preparation procedure with solid phase extraction followed, before injecting into a tandem mass spectrometer with positive mode electron spray ionisation and multiple reactant monitoring acquisition. The chromatography time was 3 min. The limit of quantitation for cortisol and cortisone in plasma was 3.75 nmol/L and linearity extended to 2000 nmol/L. The limit of quantitation for cortisol in plasma ultrafiltrate and saliva was 0.6 nmol/L. The limit of quantitation for 11-deoxycortisol and prednisolone was 5 nmol/L and for dexamethasone 1 nmol/L. The intra-assay CV was <5% and the inter-assay CV <10% for all analytes in all matrices. Comparison with an immuno-assay (IA) plasma cortisol method resulted in a regression equation of UHPLC = 0.79 × IA + 31.12 with R² = 0.960 (p < 0.0001). Comparison with a high performance liquid chromatography (HPLC) cortisol method yielded a regression equation of UHPLC = 1.06 × HPLC + 9.82, R² = 0.992 (p < 0.0001). The simultaneous measurement of endogenous and exogenous glucocorticoids contributed to patient care in cases with dexamethasone and metyrapone dynamic tests and unsuspected therapeutic glucocorticoid use.     

Relationship between change in core temperature and change in cortisol and TNFα during exercise

The combined thermal load created by exercise and a hot environment is associated with an exaggerated core temperature response. The elevated core temperature is believed to increase the total stress of the exercise. Increased stress during exercise has been associated with increased levels of cortisol. The association of cortisol with increased inflammatory responses following exercise in the heat is equivocal. Thus, the purpose of the current investigation was to explore the relationship between increases in rectal temperature (T_re) and TNFα and cortisol. To induce T_re changes, 8 male subjects (mean±SD, age=23.6±2 yr, VO₂max=52.8±3.7 mL/kg/min, BMI=24.2±1.9) participated in two 40 min trials of cycle ergometry at 65% of VO₂peak immersed to chest level in cool (25 °C) and warm (38.5 °C) water. T_re was monitored throughout each trial, with blood samples taken immediately pre and post of each trial. Neither cortisol nor TNFα changed significantly during exercise in the cool water; however, in the warm trial, both cortisol and TNFα significantly increased (p<0.004). Concordance correlations (R_c) between Δ cortisol and Δ TNFα indicated a strong but non-significant correlation (R_c=0.833, p=0.135). In conclusion, changes in core temperature may be impacting the relationship between exercise induced changes in cortisol and TNFα. Therefore, acute moderate-intensity exercise (40 min or less) in warm water impacts the stress and inflammatory response. Understanding this is important because exercise load may need to be adjusted in warm and hot environments to avoid the negative effects of elevated stress and inflammation response.     

Psychosocial stressor effects on cortisol and ghrelin in emotional and non-emotional eaters: Influence of anger and shame

Food consumption in stressful situations vary as a function of individual difference factors (e.g., emotional vs. non-emotional eating), and may be related to hormonal responses elicited by the stressful event. These hormonal responses may be tied to specific emotions elicited by the stressful event. The present investigation examined the emotional and hormonal (cortisol, ghrelin) responses of high and low emotional eaters following a laboratory stressor (Trier Social Stress Test; TSST). Women (n = 48) either high or low in emotional eating status were tested in a TSST or served as controls during which blood samples were taken for analysis of cortisol and ghrelin, both of which have been implicated in eating and in response to stressors. The TSST promoted elevated cortisol levels, being somewhat more pronounced in emotional than in non-emotional eaters. Both shame and anger were provoked by the TSST, and although both these emotions were correlated with cortisol levels, only anger significantly mediated the relationship between the stressor and cortisol levels. As well, baseline ghrelin levels in low emotional eaters exceeded that of high emotional eaters, and increased moderately in response to the stressor situation, irrespective of emotional eating status. Interestingly, when provided with food, ghrelin levels declined in the non-emotional eaters, but not in emotional eaters. The possibility is offered that the lack of a decline of ghrelin in emotional eaters may sustain eating in these individuals.     

Changes in plasma IL-1beta, TNF-alpha and IL-6 after total hip replacement surgery in general or regional anaesthesia

Different anaesthetic methods influence the neuro-immuno-endocrine biologic responses to surgery and may thus possibly interfere with the postoperative course and development of complications. The neuroendocrine system is closely related to the cytokine network. In this study, the effects of general anaesthesia (n=6) and regional spinal/epidural anaesthesia (n=6) on the cytokine response (IL-1beta, TNFalpha, IL-6) to uncemented total hip replacement surgery were evaluated. The postoperative clinical course was uneventful in every case. In both groups, only very low values of plasma IL-beta were measured perioperatively, whereas plasma IL-6 increased postoperatively with peak values 4 h after surgery. The changes in plasma TNF-alpha were not significant. No significant differences in plasma TNF-alpha or IL-6 were found between patients operated in general or in regional anaesthesia. This suggests minor influence of plasma cytokines on the possible beneficial effects of regional anaesthesia on the clinical course after surgery in low risk patients. There were slightly higher TNF-alpha and IL-6 levels after the operation and significantly lower cortisol levels during the operation in the regional anaesthesia group compared to the general anaesthesia group, giving rise to a significant inverse correlation between peak values of IL-6 and peak values of cortisol. This supports the theory that after surgery the inhibitory effect of cortisol on monocyte cytokine production overrides adrenergic stimulation.     

11β-Hydroxysteroid dehydrogenase 1 contributes to the pro-inflammatory response of keratinocytes

The endogenous glucocorticoid, cortisol, is released from the adrenal gland in response to various stress stimuli. Extra-adrenal cortisol production has recently been reported to occur in various tissues. Skin is known to synthesize cortisol through a de novo pathway and through an activating enzyme. The enzyme that catalyzes the intracellular conversion of hormonally-inactive cortisone into active cortisol is 11β-hydroxysteroid dehydrogenase 1 (11β-HSD1). We recently reported that 11β-HSD1 is expressed in normal human epidermal keratinocytes (NHEKs) and negatively regulates proliferation of NHEKs. In this study, we investigated the role of 11β-HSD1 in skin inflammation. Expression of 11β-HSD1 was induced by UV-B irradiation and in response to the pro-inflammatory cytokines, IL-1β and TNFα. Increased cortisol concentrations in culture media also increased in response to these stimuli. To investigate the function of increased 11β-HSD1 in response to pro-inflammatory cytokines, we knocked down 11β-HSD1 by transfecting siRNA. Production of IL-6 and IL-8 in response to IL-1β or TNFα stimulation was attenuated in NHEKs transfected with si11β-HSD1 compared with control cells. In addition, IL-1β-induced IL-6 production was enhanced in cultures containing 1 × 10⁻¹³ M cortisol, whereas 1 × 10⁻⁵ M cortisol attenuated production of IL-6. Thus, cortisol showed immunostimulatory and immunosuppressive activities depending on its concentration. Our results indicate that 11β-HSD1 expression is increased by various stimuli. Thus, regulation of cytosolic cortisol concentrations by 11β-HSD1 appears to modulate expression of inflammatory cytokines in NHEKs.     

Cortisol release and heart rate variability in horses during road transport

Based on plasma cortisol concentrations it is widely accepted that transport is stressful to horses. So far, cortisol release during transport has not been evaluated in depth by non-invasive techniques such as analysis of salivary cortisol and faecal cortisol metabolites. Transport also causes changes in heart rate and heart rate variability (HRV). In this study, salivary cortisol, faecal cortisol metabolites, heart rate and HRV in horses transported by road for short (one and 3.5 h) and medium duration (8 h) were determined. With the onset of transport, salivary cortisol increased immediately but highest concentrations were measured towards the end of transport (4.1 ± 1.6, 4.5 ± 2.6, 6.5 ± 1.8 ng/ml in horses transported for one, 3.5 and 8 h, respectively). Faecal cortisol metabolite concentrations did not change during transport, but 1 day after transport for 3.5 and 8 h had increased significantly (p < 0.01), reflecting intestinal passage time. Compared to salivary cortisol, changes in faecal cortisol metabolites were less pronounced. Heart rate increased and beat-to-beat (RR) interval decreased (p < 0.05) with the onset of transport. Standard deviation of heart rate increased while root mean square of successive RR differences (RMSSD) decreased in horses transported for 3.5 (from 74 ± 5 to 45 ± 6 ms) and 8 h (from 89.7 ± 7 to 59 ± 7 ms), indicating a reduction in vagal tone. In conclusion, transport of horses over short and medium distances leads to increased cortisol release and changes in heart rate and HRV indicative of stress. The degree of these changes is related to the duration of transport. Salivary cortisol is a sensitive parameter to detect transient changes in cortisol release.     

The effect of zoo visitors on the behaviour and faecal cortisol of the Mexican wolf (Canis lupus baileyi)

The effect of zoo visitors on the behaviour and physiological responses of the Mexican wolf has not been documented but is worthy of investigation since it is a critically endangered native species and most of the population live in zoos. The effect of the number of visitors was assessed in 12 wolves (five males and seven females) in three zoos of central Mexico. Each wolf was observed for a total of 72 h on Saturdays, Sundays, Mondays and Tuesdays (6 h/day) during a three-week period. Scan sampling was used to obtain individual time budgets. A faecal sample was collected on the following morning of each observation day from 10 wolves to determine cortisol level (RIA). A repeated measures analysis of variance revealed that there was an effect of day on the proportion of time lying (P < 0.01), eating (P < 0.01) and in locomotion (P < 0.01). Wolves spent less time lying on Saturdays and Sundays (0.2980 ± 0.012 and 0.3266 ± 0.012, respectively) than on Mondays and Tuesdays (0.3821 ± 0.012 and 0.4075 ± 0.012, respectively), as well as less time eating on weekend days (Saturdays 0.1214 ± 0.015, Sundays 0.0556 ± 0.015) than in the other 2 days (Mondays 0.816 ± 0.015 and Tuesdays 0.0915 ± 0.015). Mean faecal cortisol was 203.7 ng/g of DM and ranged from 113.10 to 314.48 ng/g. There was an effect of day on the faecal cortisol response (P < 0.01) with wolves having higher cortisol response on Sundays (242.50 ng/g ± 8.48) compared to the other 3 days (Saturdays 192.71 ± 8.48, Mondays 170.73 ng/g ± 7.80, Tuesdays 183.82 ng/g ± 8.48). This is the first study that measures faecal cortisol of Mexican wolves. These results indicate that the amount of visitors in zoos influence the behaviour and adrenal activity of these animals which could be undesirable for ex situ conservation efforts of this endangered species.