Determination Of Serum Cortisol Research Articles

Analysis of microsamples by miniaturized magnetic-based pipette tip microextraction: determination of free cortisol in serum and urine from very low birth weight preterm newborns.

Miniaturized magnetic-based pipette tip microextraction is presented as a sample preparation approach for microsamples. It involves quick dispersion of a diminutive amount of a magnetic sorbent material in a low-volume sample (10 μL) to entrap the target analytes. Next, the dispersion is aspirated using a (semi)automatic pipette through a pipette tip with a small cubic neodymium magnet inside, which retrieves the magnetic sorbent containing the analytes. After discarding the rest of the sample, the sorbent is properly rinsed by aspirating/dispensing deionized water, and then, the analytes are eluted by aspirating/dispensing an appropriate solvent. This approach was employed for the determination of free cortisol in serum and urine from very low birth weight preterm newborns, a vulnerable patient group who present low availability for sampling biological fluids. A magnetic immunosorbent made of a cortisol antibody was employed for the selective extraction, followed by liquid chromatography-tandem mass spectrometry. Good analytical features were obtained, such as limits of detection and quantification of 0.08 and 0.27 ng mL-1, respectively, linearity up to 50 ng mL-1 (R2 > 0.999), RSD values under 15% and relative recoveries between 91 and 111%. The cross-reactivity with other glucocorticoids (i.e., cortisone and prednisolone) was evaluated to show the selectivity of the extraction. Finally, the method applicability was demonstrated towards the determination of free cortisol in the serum and urine samples from low birth weight preterm newborns.

An Immunosensor for the Determination of Cortisol in Serum and Saliva by Array SPRi.

Cortisol is a hormone which plays an essential role in the immune, endocrine, cardiovascular, renal and skeletal systems. Its level increases in response to stress, illness, injury or exhaustion, and it is therefore a significant diagnostic biomarker of stress. An immunosensor for the determination of cortisol by SPRi array was developed. The receptive part of the immunosensor is mouse monoclonal antibody against cortisol, immobilized via cysteamine linker. The optimum pH of the immunosensor is 7.4, and the optimum concentration of the antibody is 50 ng mL-1. The immunosensor is specific for cortisol, and its linear response ranges from 0.20 ng mL-1 (LOQ) to 8 ng mL-1. The precision of the determination was between 3.1% and 3.3%, and the recovery between 99% and 102%. The immunosensor was validated by simultaneous determination of cortisol in serum and saliva samples by a standard method, with good agreement between the results.

The protective role of iodine and MS-222 against stress response and bacterial infections during Nile tilapia (Oreochromis niloticus) transportation

Fish transportation is a regular farming and experimental practice that mainly results in stress conditions that may extend to induce a high mortality rate. In the present experimental simulation, Oreochromis niloticus were transported for 2 h and maintained for another 6 h in transportation containers to assess the effect of tranquilization with tricaine methanesulfonate (MS-222) and/or the addition of iodine on mortality rate and restoring the normal physiological functions. Experimental fish were divided into four groups: group 1 (control) was transported in farm water without MS-222 or iodine, group (2) was transported in farm water supplemented with 40 mg/L of MS-222, fish in group (3) were transported in farm water supplemented with 40 mg/L of MS-222 + 10 ppm iodine, and fish in group (4) were transported in farm water containing 10 ppm iodine. Blood samples were collected for the determination of serum cortisol and glucose, while skin mucus was collected for assaying lysozyme, peroxidase and antibacterial activity; cumulative mortality rate; and food reflexes which were assessed at 0 h, 1 h, 6 h, 48 h, 7 days and 14 days post 2 h transportation. The results indicated that fish tranquilization with MS-222 and/or treatment with iodine mitigated the stress condition associated with transportation and accelerate restoration of the normal physiological and immunological status. So, it was recommended to use the MS-222 as a tranquillizer together with iodine as an antiseptic during transportation of O. niloticus.

Immune Functions Alterations Due to Racing Stress in Thoroughbred Horses.

Simple SummaryRacehorses are under constant stress when training and during competitions. It is known that high levels of cortisol, the hormone responsible for stress regulation, can impact the immune system. Hence, the importance of assessing the immune cells, blood components and cortisol during different times of athlete horses’ routines, including races. This research verified the impact of training and racing on the immune function of Thoroughbreds’ trough phagocytosis and oxidative neutrophil burst assays, serum cortisol determination, erythrocytes apoptosis, lymphoproliferation, and blood count analyses. Racehorses are constantly exposed to stress. Aiming to verify the state of blood components and cortisol alterations during their routine and after races, phagocytosis and oxidative neutrophil burst assays, serum cortisol determination, erythrocytes apoptosis evaluation, lymphoproliferation assays, and blood count tests were performed in thirty Thoroughbred racehorses, which were divided in two groups. The samples were taken right after races (moment 0 d), during rest periods (−11 d, +1 d, +3 d), and after training (−8, +2, +5). In both groups, the phagocytosis showed a decrease in percentage and intensity immediately after the race when comparing samples collected during rest or training periods. In the mean values of oxidative burst on samples collected immediately after the race, group I animals demonstrated a decrease (524.2 ± 248.9) when compared with those samples collected in other moments. No significant differences were found between the results of different moments regarding the apoptotic cells and lymphoproliferation assays. The mean values of serum cortisol levels were increased immediately after racing. There was an increase in the percentage of neutrophils found immediately after the race. It was possible to conclude that, although a transient reduction was found in the number of neutrophils, the horses’ adaptive function was not affected.

Short communication: Is it better to calve alone or in groups? A pilot study

In this pilot study, 35 multiparous Holstein cows (3.2 ± 1.16 lactations) were allocated to either a treatment consisting of cows calving in isolation or to a treatment consisting of cows calving in groups. Cows were moved 5–7 d before expected calving date to a maternity barn with two individual 25-m2 pens (SINGLE; n = 15) or to a maternity barn with 2 pens (250 m2) holding 8–10 cows (GROUP; n = 20). All cows were fed the same dry-cow and lactating-cow ration before and after calving, respectively. Incidence of retained placenta (RP), metritis (MTR), and clinical mastitis (MAST) was recorded for each cow. Cows were video-recorded and calving duration and time interacting with the newborn calf was assessed during the first 4 h after parturition. Individual milk production was recorded daily during the first 60 DIM. The proportion of cows that reached 250 DIM was also recorded. Lastly, a blood sample was obtained from the coccygeal vessels at 12 ± 2 h after calving to determine serum cortisol, insulin, and glucose concentrations. There was no difference in calving duration (94.2 ± 14.6 min) or time interacting with the calf (70.4 ± 7.1 min) between treatments. Serum cortisol concentrations were greater in SINGLE (2.25 ± 0.30 ng/ml) than in GROUP (1.20 ± 0.30 ng/ml) cows. Milk production during the first 60 DIM (2,751± 109 kg) did not differ between treatments. Cows on SINGLE tended to have a greater risk (odds ratio = 5.0) of incurring RP and had a greater risk (odds ratio = 10.6) of suffering from MTR than GROUP cows. Only 73% of the SINGLE cows reached 250 DIM, whereas 95% of the GROUP cows reached 250 DIM; thus, GROUP cows had 38 greater odds of reaching 250 DIM than SINGLE cows. It is concluded that cows kept in isolation for about 1 week before calving tend to have more RP, have greater serum cortisol concentration after calving, experience a greater incidence of MTR, and are less likely to reach 250 DIM compared with cows moved to groups about 1 week before calving.

Validation study of canine serum cortisol measurement with the Immulite 2000 Xpi cortisol immunoassay.

We report the results of validation of canine serum cortisol determination with the Immulite 2000 Xpi cortisol immunoassay (Siemens), with characterization of precision (CV), accuracy (spiking-recovery [SR] bias), and observed total error (TEo = bias + 2CV) across the reportable range, specifically at the most common interpretation thresholds for dynamic testing. Imprecision increased at increasing rate with decreasing serum cortisol concentration and bias was low, resulting in increasing TEo with decreasing serum cortisol concentration. Inter-laboratory comparison study allowed for determination of range-based bias (RB) and average bias (AB). At 38.6 and 552 nmol/L (1.4 and 20 μg/dL), between-run CV was 10% and 7.5%, respectively, and TEo ~30% and ~20%, respectively (TEo remained similar regardless of the considered bias: SR, RB, or AB). These analytical performance parameters should be considered in the interpretation of results and for future expert consensus discussions to determine recommendations for allowable total error (TEa). Importantly, the commonly used thresholds for interpretation of results were determined ~40 y ago with different methods of measurements and computation, hence updating is desirable. Quality control material (QCM) had between-run imprecision of 4% for QCM1 and 7% for QCM2; the bias was minimal for both levels. Acceptable QC rules are heavily dependent on the desired TEa for the QCM system (TEaQCM), itself limited by the desired clinical TEa. At low TEaQCM (20-33%), almost no rules were acceptable, whereas at high TEaQCM (50%), almost all rules were acceptable; further investigation is needed to determine which TEaQCM can be guaranteed by simple QC rules.

Arrival cortisol measurement in veal calves and its association with body weight, protein fractions, animal health and performance

Male dairy calves are exposed to an accumulation of transport, social and environmental stressors while transferred to fattening units. As a consequence, calves show high cortisol concentrations upon arrival at the veal facility. Whether cortisol levels as measured on arrival can be associated with animal health, welfare and production results is currently unknown.The first objective of this prospective cohort study was to determine possible associations of arrival serum cortisol concentration with health and production variables of veal calves and other arrival predictors like body weight and γ-globulin concentration. The second aim was to investigate potential clustering of arrival risk factors in veal calves for developing bovine respiratory disease (BRD) based on arrival body weight, serum cortisol concentration, total protein and protein fractions. In total, 105 male Holstein calves from two consecutive production cycles in a single, commercial white veal farm were blood sampled directly at arrival on the farm to determine serum cortisol, total protein and protein fractions. All calves were weighed the day after arrival and clinical signs, average daily weight gain (ADG) and carcass weight were collected. Also, all calves of both production cohorts were repeatedly examined by thoracic ultrasonography at the onset of group respiratory disease symptoms (2−3 weeks after arrival) and four weeks later. Linear and logistic mixed models together with k-means clustering were used for statistical analyses.Calves showed on average high, but individually variable serum cortisol concentrations (mean value = 96.6 ng/mL ± standard deviation (SD) = 48.8; Range (R) = 50.0−317 ng/mL). Arrival cortisol, body weight and γ-globulin content were not significantly associated. Serum cortisol and albumin concentrations at arrival were associated with chronic, unresponsive pneumonia. For each increase of serum cortisol concentration by 10 ng/mL, the odds for lung consolidation of ≥1 cm in depth at the second ultrasonography increased, odds ratio (OR) = 1.03 (95 % confidence interval (CI) = 1.01–1.06; P < 0.050). For every decrease in serum albumin concentration by 1g/L, the OR for developing pneumonia was 1.23 (95 % CI= 1.04–1.46; P < 0.015). Additionally, two clusters of calves were identified based upon arrival status: a low risk cluster with below average stress-induced cortisol values, above average body weight, no acute phase response (APR) and no failure of passive immune transfer (FPIT), and a high risk cluster with above average cortisol values, below average body weight, APR and FPIT. High risk calves had higher odds for developing clinical BRD (OR= 3.88 (95 % CI=1.20−12.53; P < 0.020)) and lung consolidation ≥1 cm in depth at week 6−7 after arrival (OR= 3.93 (95 % CI=1.34−11.53; P < 0.013)).Avoiding high cortisol levels upon arrival of calves is important for animal welfare, but also for reduction of the prevalence of chronic, unresponsive pneumonia and the associated need for (repeated) antimicrobial treatment and production losses.

Is ECLIA Serum Cortisol Concentration Measurement, an Accurate Indicator of Pain Severity in Dogs with Locomotor Pain?

Simple SummarySerum cortisol level reflects the activity of stress axis, ethological alterations, acute and chronic pain, life quality, or psychogenic stress. Although it is stated that stress can produce a measurable influence on the cortisol level, a certified value of this pain biomarker in dogs was not generally accepted yet. This interdisciplinary research emerged from the need for information in this field since not many studies were focused mainly on comparative analysis. We consider this field as a hot topic with various possible applications. Our survey is a methodological study within the fields of behavior and veterinary sciences, being relevant for the dog’s pain assessment. Results are having the guarantee of the high standard analysis of serum specimens, and using the updated Cortisol assay, with serum cortisol determined by Electrochemiluminescence immunoassay. We consider that our work could refresh the information in this field. Yet, an area of interest, specific pathology and pain particularities in a dog, being studied more and more in the last decade. What we can say is that serum cortisol limits cannot be adopted as a single and accurate pain marker in dog species, our study confirming these values, as non-conclusive for the assessment of the real pain levels in the dogs.The purpose of determining serum cortisol level is to reflect the activity of stress axis, ethological alterations, acute and chronic pain, life quality, or psychogenic stress. Although it is stated that stress can produce a measurable influence on the cortisol level, a certified value of this pain biomarker in dogs was not generally accepted yet. This study aimed to investigate if serum cortisol measured follows allopathic treatments only, or it is associated with physiotherapy, point out pain level in dogs with orthopedic disease, which could reveal the healing progress. The diagnostic identified: hip dysplasia, cranial cruciate ligament rupture, or intervertebral disc disease. Ortolani and Barden tests, together with clinical examination, drawer sign, and tibia compression test, were done in dogs exhibiting postures, and motion alteration, and X-Ray confirmed. A total of 30 dogs were grouped in healthy (n = 10) and pain groups (n = 20), the blood sampling is done at the beginning of the investigation, and after ten days of the study. Dogs were handled in two ways: G1—treated with Nonsteroidal Anti-Inflammatory Drugs (NSAIDs) only and respectively, G2—by therapy and physiotherapy. The analysis was performed on a Roche Cobas Analyzer (Roche, USA), serum cortisol being determined by Electrochemiluminescence immunoassay (ECLIA), and statistics using ANOVA, following Tukey’s Multiple Comparison Test. The results revealed that, out of ten specimens in the Control group, nine were within the normal limits: 5–65 ng × mL−1 (24.76 ± 19.48678), and one sample under the set limit. In G1, it was observed that the plasmatic P1 values were below the levels of P2, in six situations. In G2, although the status of all subjects improved radically with the removal or evident reduction of pain, confirmed clinically and imagistically, the P2 values in five dogs were higher than the initial P1 values, and in contradiction with the observed clinical reality. Comparing results, the mean difference in G1 was 0.41, and in G2 = 2.54, with an SD for G1 = 13.38, and G2 = 16.66, registering moderate development. Standard deviation illustrated that the values of treated groups were highly spread throughout the interval, and the serum cortisol assay did not generate significant statistical differences between groups in our case. This inferred the doubt whether the used detection method or values registered correctly indicates the pain levels in dog species.

BIOMEDICAL INDICATORS OF PRE-COMPETITIVE STRESS IN FEMALE AND MALE JUDO PLAYERS

The aim of the present paper is to monitor changes of selected mental and physiological stress indicators in both female and male sportsmen in the subsequent days prior to an International Judo Tournament in Poland and on the competition day. The study included 9 men and 9 women, participants of the training camp that ended with a tournament. Every morning, on the three consecutive days preceding the competition (day -3, -2, -1) and on the competition day (day 0) capillary blood was sampled to determine serum cortisol (C) and testosterone (T) levels, then the subjects assessed their night sleep quality (SQ) with the use of a 5-point Likert scale, and perceived anxiety using the State-Trait Anxiety Inventory questionnaire by Spielberger (STAI - Polish version). SQ insignificantly deteriorated, and C and anxiety levels increased with the time to event. The inter-group analysis for the entire period showed higher scores in average anxiety and higher average level of C in women than men. The relative changes in these biomarkers could indicate higher pre-competitive stress in women, however without taking into account the sports class of the opponents individually for each of the groups one cannot imply gender-specific susceptibility to stress as a general principle.

Serum cortisol and thyroid hormone concentrations and survival in foals born from mares with experimentally induced ascending placentitis.

BackgroundThere are few publications on occurrence of nonthyroidal illness syndrome in foals and on the prognostic value of cortisol and thyroid hormone (TH) concentrations in newborn foals.ObjectivesTo determine serum cortisol and TH concentrations (total and free thyroxine: T4 and FT4; total and free triiodothyronine: T3 and FT3) in foals born from mares with placentitis, to determine their association with survival, and their use as prognostic markers.AnimalsA cohort of 29 newborn foals comprising 5 Control, 14 Low‐risk, and 10 Sick foals were evaluated over the first week of life.MethodsIn this prospective study foals born to mares with experimentally‐induced placentitis were assigned to Low‐risk or Sick groups while foals born to control mares were classified as Control based on clinical findings. Foals were also classified as Term (n = 13), Dysmature (n = 7), or Premature (n = 9), and survival rate was recorded. Serum cortisol and TH hormone concentrations were measured at 0, 12, 24, 48, and 168 hours of life.ResultsSick non‐surviving foals had lower (P < .05) T3 : cortisol ratio at 12 (3.68 ± 1.06 versus 18.58 ± 2.78), 24 (5.47 ± 2.34 versus 23.40 ± 3.82), and 48 (10.47 ± 6.29 versus 26.6 ± 2.90) hours of life when compared to Sick surviving foals and lower (P < .05) T4 : cortisol ratio at 12 (75.12 ± 21.71 versus 414.47 ± 58.47) and 24 hours (127.83 ± 55.21 versus 430.87 ± 80.31) after birth than Sick surviving foals.Conclusion and Clinical ImportancePlacental infections can impair fetal thyroid function. Low T3 : cortisol and T4 : cortisol ratios seem to be good prognostic markers in newborn foals.

Male High School Students Express Higher Blood Cortisol Levels During Examination Periods than Female High School Students.

The academic formation of an individual with good study opportunities in Mexico begins approximately at 3 years of age and remains constant until graduation from high school. At this stage, there are learning periods, that include classes and homework, and evaluation periods, that covers exams and class presentations. Evaluation periods concentrate two or three daily exams during one week. The present project was designed to determine through cortisol concentrations if there are differences in the expression of stress between males and females in high school in a private school in the city of Puebla, Mexico. We recruited 14 female and 13 male adolescent volunteers between the ages of 17 and 18. We obtained two blood samples, the first one was taken during the class period and the second one was taken on the week of final evaluations of the semester. The blood was processed according to the instructions of an ELISA Bio-cortisol (MEXLAB) kit in order to determine serum cortisol concentrations, and according to a Bio-glucose (MEXLAB) kit to determine serum glucose concentrations. The results show that males have higher blood cortisol levels during the class and examination periods compared to females. We also show that cortisol levels in males are significantly higher during the examination period than during the class period; whereas females did not show differences in cortisol levels between periods. In the case of glucose levels, we observed differences related to sex but not differences related to the evaluated periods. These data suggest that school activities at the high school level generate more stress in males than in females.

SAT-355 Comparison of Two Serum Cortisol Immunoassays versus Liquid Chromatography: Tandem Mass Spectrometry (LC-MS/MS)

Accurate measurement of serum cortisol is required to diagnose and treat adrenal disorders. The most widely used methods in clinical laboratories for serum cortisol measurement are immunoassays which are offered in numerous commercial kits and on automated platforms. The LC-MS/MS is the gold-standard method to quantify serum steroids due its highest specificity, however it is a laborious methodology. Objective: to compare serum cortisol levels measured by two immunoassays versus LC-MS/MS. Methods: 35 routine serum samples (range: 1.1 to 50.0 ug/dL) were measured by Access Cortisol in Unicel DxI800 (Beckman Coulter), Architect Cortisol in i2000 Architect (Abbott) and LC-MS/MS in Xevo-TQ-S (Waters). Results were compared by Deming regression analysis and mean bias were used to describe the relationship between LC-MS/MS and Access and Abbott immunoassays, respectively. Results: Beckman and Abbott immunoassays showed similar cortisol concentrations (mean bias: 0.366%). Both immunoassays showed close agreement with LC-MS/MS results. Deming regression analysis showed a slope of 0.99 (0.85 to 1.134) an intercept of 1.334 (-1.345 to 4.014) in LC-MS/MS vs Beckman comparison; a slope of 1.008 (0.906 to 1.110) an intercept of 1.164 (-0.766 to 3.094) in LC-MS/MS vs Abbott comparison. The mean positive bias of Beckman and Abbott versus LC-MS/MS were 8.38% e 8.78%, respectively. Conclusion:Both Beckman and Abbott Cortisol immunoassays presented similar results in comparison of cortisol LC-MS/MS measurement, which evidence that these assays are suitable and reasonable methods for routine serum cortisol determination.

Comparing hair cortisol concentrations from various body sites and serum cortisol in Holstein lactating cows and heifers during thermal comfort zone

Measuring cortisol in hair has recently been established as a reliable physiological index to evaluate chronic stress conditions in humans and many animal species. The aim of this study was to determine serum cortisol and the concentrations of hair cortisol from various body sites of lactating Holstein cows and heifers during thermal comfort zone. Forty-seven multiparous lactating Holstein cows (average milk yield 37.5 ± 2.3 kgd−1, days in milk = 110 ± 47 days, body weight = 623 ± 75 kg) and 23 Holstein heifers (10-12 months of age, body weight = 258 ± 20 kg) were used in this study. Blood samples were collected on days (d) 0, 30, and 60 of the study. Hair samples were harvested from the forehead, withers, and rump sites of the animals. Data for temperature-humidity index (average THI = 69.3 ± 1.2) represented the threshold environmental conditions during the experiment. Serum cortisol concentration was higher on day 0 of the study and lower on day 30 and day 60 of sampling in lactating Holstein cows and heifers (P < 0.05). However, no differences in serum cortisol concentrations were observed in the second (day 30) and the third (day 60) samples in heifers and lactating cows (P < 0.05). No differences were found in hair cortisol concentrations among various sampling sites in lactating cows and heifers (P > 0.05). Conclusions drawn indicate that the distribution of cortisol into the hair shaft does not depend on hair sampling sites, and so that of the hair cut for cortisol analysis can be collected from any of the three body sites.

Can cortisol levels predict the severity of acute whiplash-associated disorders?

The exact underlying mechanism of whiplash-associated disorders still remains obscure. Central sensitization of the brain to painful stimulus and disturbances in the hypothalamic-pituitary-adrenal axis has been suggested to contribute to the development of whiplash-associated disorders. Although cortisol is a well-known factor in the acute stress response and its effects on chronic pain sensation were studied, information is lacking regarding the relation between acute phase cortisol concentrations and the intensity of whiplash-associated disorders. The aim of this prospective observational study was to investigate the relationship between acute serum cortisol concentrations and the severity of whiplash-associated disorders. 55 patients enrolled in the study and they answered a pertinent questionnaire. A blood sample was drawn to determine serum cortisol concentration. The mean cortisol concentration of the whiplash-associated disorder score 2-3 patients was significantly lower compared to the whiplash-associated disorder score 1 patients, 9.5 ± 6.9 vs. 13.22 ± 8.3µg% (p = 0.02). The mean cortisol concentrations increased significantly from mild through moderate to serious grade of severity of accident as perceived by the patient, 9.64 ± 4.82, 11.59 ± 6.85, 17.39 ± 12.1µg% (p = 0.02). The study supports the possibility that cortisol plays a role in the development of whiplash-associated disorders. Low or relatively low cortisol concentrations might be associated with more severe forms of the disorder.

0300 STRESS SYSTEM DYSREGULATION IN INSOMNIA DISORDER

Insomnia disorder (ID) is highly co-morbid with chronic pain conditions. Dysregulation of the hypothalamus-pituitary adrenal (HPA) stress system suggests a contribution to both ID and chronic pain. We investigated HPA system integrity in individuals with ID, indicated by measures of resting cortisol levels and IL-6 expression in monocytes, glucocorticoid (GC) sensitivity (i.e., sensitivity of cells to the counter-inflammatory signal cortisol), and reactivity of cortisol to a repeated challenge. We hypothesized deterioration of HPA system integrity may represent a mechanistic pathway by which ID increases chronic pain vulnerability. This is an ongoing study with N=19 participants (ages 18–49). Seven participants (1 male) diagnosed of insomnia disorder (ID) based on DSM-V criteria and 12 healthy controls (HC;3 male) completed an overnight polysomnographic (PSG) sleep recording in the Clinical Research Center. The following morning, resting blood samples were collected for measurements of basal serum cortisol levels, IL-6 positive monocytes, and GC sensitivity of monocytes as assessed by flow cytometry. To determine serum cortisol reactivity, participants completed a cold pressor test (CPT) challenge three times in succession (1300,1430,1600) by placing their hand in a temperature-controlled cold water bath for at least one minute. Blood was drawn 20 and 50 minutes after each CPT to measure post-challenge cortisol levels. ID participants had significantly shorter PSG sleep duration (ID:341 ± 110 min; HC:422 ± 31 min). Basal cortisol levels, IL-6 positive monocytes and GC sensitivity did not differ between groups, but cortisol levels in the ID group were significantly higher 20 min (ID:14.24 ± 3.34μg/dL; HC:10.86 ± 2.38μg/dL) and 50 min (ID:11.46 ± 2.40μg/dL; HC:8.99 ± 2.04μg/dL) after the first CPT challenge, and 20 min after the second CPT challenge (ID:11.24 ± 3.59; HC:7.92 ± 1.59μg/dL). While basal levels of cortisol, IL-6 expression by monocytes, and GC sensitivity did not significantly differ between ID and HC cortisol reactivity in the first and second CPT challenge was increased. These preliminary results suggest a stronger stress system responsiveness to novel challenges, which may mechanistically contribute to the relationship between ID and vulnerability to chronic pain. Investigator-initiated grant from Merck Inc., NIH/UL1 RR02758 and M01-RR-01032 from National Center for Research Resources to Harvard Clinical and Translational Science Center.

Increases in heart rate and serum cortisol concentrations in healthy dogs are positively correlated with an indoor waiting‐room environment

Few studies have investigated the effect of veterinary clinical procedures on the welfare of dogs, with specific emphasis on the veterinary practice environment. Clinicopathologic variables have also not been assessed in these potentially stressful situations. Similar to human clinical studies, the veterinary clinical waiting room could present a significant stress factor for dogs. The present study was designed to investigate the effect of waiting-room environment on serum cortisol and glucose alterations as well as heart rate in privately owned healthy dogs. The clinical trial included 24 healthy dogs that were divided into 2 groups: the clinical waiting-room group (A) and the control group (B) that waited outside in a garden. During the entire experiment, 18 dogs (9 dogs per group) were monitored with a human heart rate monitor fastened around the chest. After 20 minutes of waiting, blood samples were collected from all of the dogs (24 dogs) to determine serum cortisol concentration. Serum cortisol concentration and mean, maximum, and minimum heart rate were significantly higher in group A compared with group B, but there was no statistical difference in serum glucose concentrations between the 2 study groups. Results of this study suggest that the waiting room is a potentially stressful situation for dogs in clinical veterinary practice, when compared with a garden, based on the assessment of adrenal cortex function and heart rate evaluation.

Effects of music therapy on pain and anxiety in patients undergoing port catheter placement procedure

Patients scheduled to invasive medical procedures experience high levels of anxiety, which may lead to increased perceptions of pain and vital sign instability throughout. To examine the effect of a music intervention (MI) on stress hormones, physiologic parameters, pain, and anxiety state before and during port catheter placement procedures (PCPPs). We conducted a prospective, randomized, controlled study in 100 oncology patients, who were randomly assigned to an MI group (n=50) or a control group (n=50). The effects of music were assessed by determination of serum cortisol and adrenocorticotropic hormone (ACTH) levels, heart and respiratory rate (HR, RR) and systolic and diastolic blood pressure (SBP, DBP), on arrival in the surgical intervention room, as well as immediately prior to and immediately after the PCPP, in both groups. Furthermore, pain and anxiety levels were identified using visual analogue scale and state-trait anxiety inventory scales. On arrival, there were no differences between the patients in terms of serum cortisol and ACTH levels, HR, RR, SBP, DBP and anxiety levels. There were significant reductions in hormone levels (p<0.05 for all), HR (p<0.001), RR (p<0.001), SBP (p<0.05) and DBP (p<0.05), immediately prior to and immediately after the PCPP in participants in the MI group compared to those in the control group. Furthermore, music led to a significant reduction in pain (p<0.05) and anxiety scores (p<0.05) in the MI group compared to control group. During invasive medical procedures, MI significantly decreases stress hormone levels, physiological parameters, acute procedural pain and anxiety.

The importance of diet choice on stress-related responses by lambs

Farm animals are commonly restricted to a reduced array of foods, like total mixed rations or pastures with low species diversity. Under these conditions, animals are less likely to satisfy their specific and changing nutrient requirements. In addition, foods and flavors eaten too frequently or in excess induce sensory-specific satiety and can cause aversions. Thus, sensory and postingestive monotony may reduce animal welfare. We hypothesized that exposure to monotonous diets, even if they are considered to be nutritionally balanced, is stressful for sheep. Twenty-four 2-month-old male Corriedale lambs were randomly assigned to two experimental groups. One group (diversity treatment, DIV) received a free choice of four-way combinations of two foods with low and two foods with high protein/energy ratios from an array of seven foods (three foods high in protein/energy ratio: soybean meal, sunflower meal, and alfalfa pellets, and four foods low in protein/energy ratio: barley grain, oat grain, milo grain, and corn grain). The other group (monotony treatment, MON) was fed a balanced ration containing all foods offered to lambs in DIV. Foods were offered in four individual buckets and exposure lasted 55 days. During exposure, feeding behavior was assessed, and blood samples were taken for a complete blood cell count and to determine serum cortisol concentration. Lambs in MON showed greater cortisol levels (31.44 vs. 19.90±3.30nmol/L [means±SEM]; P=0.025) and a greater neutrophil to lymphocyte ratio (0.37 vs. 0.26±0.05; P=0.044) than lambs in DIV. Lambs in DIV spent a lower proportion of time eating (0.38 vs. 0.49±0.02; P<0.001) and showed a greater intake rate (17.73 vs. 14.09±1.26g/min, P<0.044) than lambs in MON. They also showed a greater proportion of time lying (0.44 vs. 0.36±0.03; P=0.049) and greater activity (0.047 vs. 0.035±0.003; P=0.003) than lambs in MON. However, final body weight and the average daily weight gain were not affected by treatment (P>0.05). Our results showed that restricting lambs’ dietary breadth produced changes in blood and behavioral parameters previously shown to be indicative of stress in sheep. The importance of incorporating food choice as an alternative practice to overcome stress associated to the traditional livestock feeding management is discussed.

Evaluation of thymus morphology and serum cortisol concentration as indirect biomarkers to detect low-dose dexamethasone illegal treatment in beef cattle

BackgroundCorticosteroids are illegally used in several countries as growth promoters in veal calves and beef cattle, either alone or in association with sex steroids and β-agonists, especially at low dosages and primarily through oral administration, in order to enhance carcasses and meat quality traits. The aim of the present study is to evaluate the reliability of the histological evaluation of the thymus, as well as the serum cortisol determination, in identifying beef cattle, treated with two different dexamethasone-based growth-promoting protocols and the application of different withdrawal times before slaughter.ResultsOur findings demonstrate that low dosages of dexamethasone (DXM), administered alone or in association with clenbuterol as growth promoter in beef cattle, induce morphologic changes in the thymus, resulting in increase fat infiltration with concurrent cortical atrophy and reduction of the cortex/medulla ratio (C/M). In fact, the C/M value was significantly lower in treated animals than in control ones, with both the protocols applied. The cut off value of 0.93 for the cortex/medulla ratio resulted to be highly effective to distinguish control and treated animals. The animals treated with DXM showed inhibition of cortisol secretion during the treatment period, as well as at the slaughterhouse, 3 days after treatment suspension. The animals treated with lower doses of DXM in association with clenbuterol, showed inhibition of cortisol secretion during the treatment period, but serum cortisol concentration was restored to physiological levels at slaughterhouse, 8 days after treatment suspension.ConclusionsThe histological evaluation of thymus morphology, and particularly of the C/M may represent a valuable and reproducible method applicable to large-scale screening programs, due to the easy sampling procedures at slaughterhouse, as well as time and cost-saving of the analysis. Serum cortisol determination could be considered as an useful in vivo biomarker of dexamethasone illegal treatment in beef cattle during the fattening period, whilst it does not appear to be a good biomarker at the slaughterhouse, since the protocol of DXM administration, as well as the withdrawal period could affect the reliability of the method.

Serum cortisol and haematological, biochemical and antioxidant enzyme variables in horse blood sampled in a slaughterhouse lairage, immediately before stunning and during exsanguination

The aim of the study was to determine changes of serum cortisol and biochemical, haematological and antioxidant enzyme variables in the blood of horses sampled during the pre-slaughter period (in the lairage and in the stunning box) and during exsanguination. A total of 24 Slovenian warm-blooded horses were observed. Blood samples for determination of serum cortisol and biochemical, haematological (red blood cell count, haematocrit, haemaglobin concentration) and antioxidant enzyme (whole blood superoxide dismutase and glutathione peroxidase) variables were collected by venipuncture of the left jugular vein in the lairage pen, 60 min before stunning (lairage) and immediately before stunning (stunning box). At exsanguination, blood samples were collected from the wound at the time of jugular vein sticking. During blood collection in the lairage pen and in the stunning box, horses were gently restrained with a halter. They were stunned using a penetrating captive bolt pistol impelled by air and were bled by jugular vein sticking. Horses were physically active in the lairage pen and in the race before entering the stunning box. After stunning, the horses showed paddling movements with their legs. In horses, the plasma lactate and glucose concentrations, the serum potassium concentration, the activities of the serum muscle enzymes aspartate aminotransferase and creatine kinase, and values of most of the other biochemical (Table 1) and haematological variables (Table 2), were significantly (P < 0.05) higher at exsanguination, than in blood sampled while they were in the lairage and in the stunning box. The serum concentrations of cortisol and chloride and the activities of alanine aminotransferase and antioxidant enzymes were not significantly different between the pre-slaughter period and exsanguination. All selected blood variables were not significantly different between the lairge and the stunning box sampling time, indicating no physiological stress responses of the investigated horses to stressors, such as novelty of the pre-slaughter environment and handling, present in the slaughterhouse between the lairage and the stunning box. However, the significantly higher values, at exsanguination, for the plasma lactate and glucose concentrations, serum muscle enzyme activities and haematological variables, than during the pre-slaughter period, might partially be attributed to stimulation of the sympathetic nervous system, caused by stunning and bleeding.