Increased Cortisol Concentrations Research Articles

Elimination of fall in glucose during glucagon stimulation test does not completely abolish an increase in growth hormone and cortisol concentrations

Elimination of fall in glucose during glucagon stimulation test does not completely abolish an increase in growth hormone and cortisol concentrations

Short Road Transport and Slaughter Stress Affects the Expression Profile of Serotonin Receptors, Adrenocortical, and Hematochemical Responses in Horses.

Horse transport is considered a cause of stress in animals and is known to affect the 5-HT concentrations in both the brain and other tissues. The aim of this research was to investigate the effect of horse transportation and slaughter stress on plasma serotonin's concentration and the expression levels of the related 5-HT1B and 5-HT2A receptors in PBMCs. Furthermore, the IL-12 levels and a variety of blood parameters, including triglycerides, total cholesterol, glucose, aspartate aminotransferase, creatine phosphokinase, lactate dehydrogenase, were also considered. This research was carried out on 32 horses submitted to short road transport of 40 km to slaughter. Blood samples were collected in baseline conditions (T0) and 24 h later, after they were slaughtered (T1). The results showed a significantly increased expression of 5-HT1B and 5-HT2A receptors and a significantly decreased expression of IL-12 in PBMCs at T1 vs. T0. Furthermore, a significant increase in cortisol and glucose concentrations, and LDH activity was observed at T1. In contrast, a significantly lower circulating 5-HT concentration was observed at T1 vs. T0. These results indicate that the stress induced by transport and slaughter stimuli led to the serotoninergic system's activation, suggesting that the expression of serotonin receptors could be used as a pivotal marker of stress, with potential applications for the improvement of elective protocols to observe the guidelines relating to transported horses.

Effects of different hormonal treatments on spermatogenesis advancement in hatchery-produced greater amberjack Seriola dumerili (Risso 1810)

In earlier studies, wild-caught greater amberjack Seriola dumerili (Risso, 1810) males reared in sea cages showed gametogenesis impairment and low sperm production and quality. Here, we (a) examined if F1 hatchery-produced males reared in sea cages also exhibit reproductive dysfunctions and (b) evaluated the effects of gonadotropin releasing hormone agonist (GnRHa) administration through injections (GnRHainj) or sustained-release implants (GnRHaimpl), and human chorionic gonadotropin (hGC) injections on spermatogenesis/spermiation enhancement. Fish were given a hormone treatment just prior to the spawning season, and were transferred to land-based tanks, according to an established spawning induction protocol. Blood samples (n = 6) were obtained on Days 0, 7 and 13 after treatment. Testis samples were obtained on Days 0 (n = 4) and 13 (n = 2 per treatment). The fish prior to their transfer from the sea cages to the land-based tanks, exhibited a low gonadosomatic index, altered sex steroid hormone profile and high density of testicular apoptotic cells. After transfer to tanks, there was a general depression of sex steroid plasma levels parallel to an increase in cortisol concentrations. Despite the negative effect on steroidogenesis by the transfer from the sea, the hormonal treatments increased the number of fish from where sperm could be obtained, as well as testis growth, and reduced testicular apoptosis. Treatment with hCG resulted in the most significant changes in spermatogenesis, while GnRHaimpl appeared to induce less intense, but likely longer-lasting effects. The study indicated that F1 hatchery-produced males also exhibited reproductive dysfunctions as wild-caught captive-reared greater amberjack, and that the observed positive effects of the hormone treatments on spermiation/spermatogenesis were likely mediated by factors other than sex steroid hormones.

Prenatal cortisol exposure impairs adrenal function but not glucose metabolism in adult sheep.

Adverse environmental conditions before birth are known to programme adult metabolic and endocrine phenotypes in several species. However, whether increments in fetal cortisol concentrations of the magnitude commonly seen in these conditions can cause developmental programming remains unknown. Thus, this study investigated the outcome of physiological increases in fetal cortisol concentrations on glucose-insulin dynamics and pituitary-adrenal function in adult sheep. Compared with saline treatment, intravenous fetal cortisol infusion for 5 days in late gestation did not affect birthweight but increased lamb body weight at 1-2 weeks after birth. Adult glucose dynamics, insulin sensitivity and insulin secretion were unaffected by prenatal cortisol overexposure, assessed by glucose tolerance tests, hyperinsulinaemic-euglycaemic clamps and acute insulin administration. In contrast, prenatal cortisol infusion induced adrenal hypo-responsiveness in adulthood with significantly reduced cortisol responses to insulin-induced hypoglycaemia and exogenous adrenocorticotropic hormone (ACTH) administration relative to saline treatment. The area of adrenal cortex expressed as a percentage of the total cross-sectional area of the adult adrenal gland was also lower after prenatal cortisol than saline infusion. In adulthood, basal circulating ACTH but not cortisol concentrations were significantly higher in the cortisol than saline-treated group. The results show that cortisol overexposure before birth programmes pituitary-adrenal development with consequences for adult stress responses. Physiological variations in cortisol concentrations before birth may, therefore, have an important role in determining adult phenotypical diversity and adaptability to environmental challenges.

Identification of intestinal and fecal microbial biomarkers using a porcine social stress model.

Understanding the relationships between social stress and the gastrointestinal microbiota, and how they influence host health and performance is expected to have many scientific and commercial implementations in different species, including identification and improvement of challenges to animal welfare and health. In particular, the study of the stress impact on the gastrointestinal microbiota of pigs may be of interest as a model for human health. A porcine stress model based on repeated regrouping and reduced space allowance during the last 4 weeks of the finishing period was developed to identify stress-induced changes in the gut microbiome composition. The application of the porcine stress model resulted in a significant increase in salivary cortisol concentration over the course of the trial and decreased growth performance and appetite. The applied social stress resulted in 32 bacteria being either enriched (13) or depleted (19) in the intestine and feces. Fecal samples showed a greater number of microbial genera influenced by stress than caecum or colon samples. Our trial revealed that the opportunistic pathogens Treponema and Clostridium were enriched in colonic and fecal samples from stressed pigs. Additionally, genera such as Streptococcus, Parabacteroides, Desulfovibrio, Terrisporobacter, Marvinbryantia, and Romboutsia were found to be enriched in response to social stress. In contrast, the genera Prevotella, Faecalibacterium, Butyricicoccus, Dialister, Alloprevotella, Megasphaera, and Mitsuokella were depleted. These depleted bacteria are of great interest because they synthesize metabolites [e.g., short-chain fatty acids (SCFA), in particular, butyrate] showing beneficial health benefits due to inhibitory effects on pathogenic bacteria in different animal species. Of particular interest are Dialister and Faecalibacterium, as their depletion was identified in a human study to be associated with inferior quality of life and depression. We also revealed that some pigs were more susceptible to pathogens as indicated by large enrichments of opportunistic pathogens of Clostridium, Treponema, Streptococcus and Campylobacter. Generally, our results provide further evidence for the microbiota-gut-brain axis as indicated by an increase in cortisol concentration due to social stress regulated by the hypothalamic-pituitary-adrenal axis, and a change in microbiota composition, particularly of bacteria known to be associated with pathogenicity and mental health diseases.

166 Pre-Weaned Calves Have Increased Cortisol Concentrations Post-Wildfire Smoke Exposure

Abstract The western states of the U.S., home to some of the largest cattle operations in the country, are increasingly experiencing losses related to wildfire events. Wildfires are predicted to intensify with climate change; rising temperatures, droughts, and drier climatic conditions. Although wildfires do not directly endanger most humans and animals, the resultant smoke can disperse great distances and increase disease susceptibility and death in populations removed from its origin. There is a lack of literature on how wildfire smoke exposure can affect livestock physiology, health, and productivity. Therefore, this study aimed to evaluate changes in blood markers of cattle naturally exposed to wildfire smoke. Eighteen beef-on-dairy calves (Jersey X Simmental) from a commercial farm were enrolled in this study at birth (May to June 2022). Blood samples for analysis of cortisol, acute phase proteins, and pro- and anti-inflammatory cytokines were collected before (June to August), during (September), and after smoke exposure (September and October), in a total of seven collections. Additionally, rectal temperature and health scores were collected from each calf at those time points. Air quality (AQ) data, specifically particulate matter (PM2.5; EPA standard for AQ; 35 µg/m3) near the study location (Vale, OR), was obtained from the AirNow database. For statistical analysis, calf was considered the experimental unit, and all data were analyzed using the MIXED procedure of SAS (SAS Inst. Inc., Cary, NC). Air quality before smoke exposure was considered adequate, and PM 2.5 was, on average, 4.9 µg/m3 from June to August. In September, a wildfire reduced AQ at the study location, increasing PM 2.5 to an average of 44.6 µg/m3 for four consecutive days. After the fire was contained, AQ improved, and PM2.5 dropped to 8.6 ug/m3. Blood cortisol concentrations changed over time (P < 0.001). Cortisol concentrations of samples collected before smoke exposure did not differ (P = 0.89) at 4.4, 2.0, and 1.9 ng/mL (SEM:1.3). Cortisol concentrations increased (P < 0.001) and peaked during smoke exposure, at 10.3 and 7.0 ng/mL (SEM:1.4). Immediately after smoke exposure, cortisol concentrations averaged 6.3 ng/mL, intermediate between those before and during smoke exposure (P = 0.89; SEM:1.3). Interestingly, in the last post-smoke exposure sample, cortisol concentration averaged 9.5 ng/mL, similar to concentrations during smoke exposure (P = 0.99; SEM:1.3). This increase in cortisol concentration post-smoke exposure was unexpected but likely explained by the calves having been weaned two days before this final sampling. In summary, calves naturally exposed to wildfire smoke had increased blood cortisol concentrations with concentrations similar to those observed shortly after post-weaning, a practice known to be challenging and stressful for calves.

113 Maternal Stress Influences Dam and Progeny Cortisol, Immune, and Behavior

Abstract Maternal stress exposes fetuses to increased cortisol concentrations, resulting in lifelong and multi-generational effects on physiology and behavior, especially if stress occurs during sensitive periods of development. This study aimed to validate the prenatal stress model and characterize the immune phenotype of the dam and the stress responsiveness and behavioral phenotype of her progeny. Gilts were randomly assigned to receive either hydrocortisone acetate (HCA) at mid (MID) or late (LATE) gestation for 21 days or placebo (CON). Blood or saliva samples were taken from gilts during the 21 day- treatment period. Blood samples were taken, and behavior was registered in response to weaning from the progeny. Body weights were also recorded. Data were analyzed using PROC MIXED with repeated measures (SAS 9.4). Significant treatment*day interactions occurred for salivary and plasma cortisol. The MID had the greatest salivary cortisol at 3- and 21-days post-HCA treatment and the least plasma cortisol at 7- and 21-days post-HCA compared with LATE and CON. Also, at 21 days post-HCA, there was a significant treatment effect where LATE gilts had decreased percentage of neutrophils (P = 0.012), increased percentage of lymphocytes (P = 0.012), and decreased neutrophil to lymphocyte ratio (P = 0.0007) compared with their respective controls. Plasma IgG patterns were differentially affected by HCA treatment. At 7 days post-HCA, IgG decreased among MID gilts but increased at d 21, whereas IgG increased at d 7 and then decreased at d 21 among LATE treated gilts. Colostrum IgA was greater in the CON gilts than in MID or LATE-treated gilts (P = 0.032). Among the pigs born to these gilts, plasma cortisol was similar before weaning, but 24-h post-weaning pigs from CON had greater cortisol than those from MID or LATE (P < 0.05), and leukocyte distribution in response to weaning was also affected by maternal treatment (P < 0.01). Finally, behavioral responses, including social interactions, were affected by maternal treatment (P < 0.001). Taken together, these findings imply that HCA treatment differentially affects the immune function and HPA responsiveness of the dam dependent on when stress is experienced within gestation.

Influence of Diet Changes on the Condition and Physiological State of Juvenile Sea Trout (Salmo trutta)

Abstract The aim of the study was to determine the influence of diets (factor D) and the time period (factor T) during which they were applied on the growth performance and physiological condition (blood plasma hematological and biochemical indicators of stress and immunity) in juvenile sea trout (Salmo trutta; initial body weight approximately 73 g). The diet of the fish that was used prior to the experiment (formulated feed; initial fish sample) was modified as follows: a different formulated feed (group B), mixed feed (feed B + prey fish; group B/N), prey fish exclusively (group N). The fish from group A were given the feed that was used prior to the beginning of the experiment. During the 28-day trial neither factors D nor T influenced absolute or relative fish growth rates. Factor D significantly influenced hematological indicators and leukograms, while the phagocytic index and cidal ability were determined by the time test (factor T). Myeloperoxidase (AMPO) was related significantly with the influence of factors D and T and also with the interaction of D×T. A significant decrease in AMPO was noted after two weeks of the test in groups B, N, and B/N, but after four weeks this indicator did not differ from that confirmed in the initial fish sample. The factors tested influenced stress indicators, i.e., cortisol (D and T) and glucose (D). Significant increases in cortisol (group B) and glucose (groups A, B, and B/N) concentrations were noted after two weeks of the test. After the subsequent two weeks (four weeks of the test) these indicators also stabilized. Sea trout from aquaculture readily accept prey fish; however, changes in diet cause stress reactions such as temporarily reduced immunity. Thus, the procedure for preparing sea trout stocking material for release, which is to change the feed from formulated to natural (live fish), should last approximately four weeks.

Metabolic rates in female guinea pigs during different reproductive stages

Reproduction in female mammals is characterized by major changes in steroid hormone concentrations, which can be linked to fluctuations in energy expenditure (EE). Estradiol and cortisol can increase EE and metabolic rates (MRs), but knowledge on MR changes during the estrous cycle and gestation is scarce for many species. This also applies to the domestic guinea pig, a species exhibiting an exceptional estrous cycle among rodents. In this study, MRs were measured through oxygen (O2) consumption in female guinea pigs during different reproductive stages. Mean O2 consumption over 2.5 h, resting metabolic rate (RMR, lowest and most stable O2 consumption over 3 min), body mass, fecal estrogen and progesterone, and saliva cortisol concentrations were measured in twelve female guinea pigs in a repeated measurements design during diestrus, estrus, and the second trimester of gestation. In estrus, body mass was significantly lower and estrogen and cortisol concentrations were significantly higher compared to diestrus and gestation. Mean O2 consumption and RMR both were significantly increased in estrus compared to diestrus. Additionally, a positive effect of body mass on MRs detected during diestrus and gestation was not found during estrus. Mean O2 consumption was also higher during gestation compared to diestrus, and a significant increase in cortisol concentrations during the 2.5-h MR measurement was recorded. The results indicate that estrus in guinea pigs is energetically demanding, which probably reflects catabolic effects of estrogens and cortisol that uncoupled MRs from body mass. Knowledge on the energetic requirements associated with different reproductive stages is important for future physiological and behavioral studies on female guinea pigs.

Exploring the Effect of Age on the Reproductive and Stress Physiology of Octopus bimaculoides Using Dermal Hormones.

Our goal was to validate the use of dermal swabs to evaluate both reproductive and stress physiology in the California two-spot octopus, Octopus bimaculoides. Our objectives were to (1) use dermal swabs to evaluate glucocorticoids and reproductive hormones of O. bimaculoides; (2) determine the influence of life stage on hormone production (glucocorticoids in all individuals; testosterone, estrogen, and progesterone in females; and testosterone in males) of reproductive (n = 4) and senescent (n = 8) individuals to determine the effect of age on hormonal patterns; and (3) determine whether these hormones change significantly in response to an acute stressor. For the stress test, individuals were first swabbed for a baseline and then chased around the aquarium with a net for 5 min. Afterward, individuals were swabbed for 2 h at 15 min intervals to compare to the pre-stress test swab. Reproductive individuals responded to the stressor with a 2-fold increase in dermal cortisol concentrations at 15 and 90 min. Six of the eight senescent individuals did not produce a 2-fold increase in dermal cortisol concentrations. Reproductive individuals had significantly higher sex hormone concentrations compared to senescent individuals (progesterone and estradiol measured in females, and testosterone for both sexes). After the stressor, only reproductive males produced a 2-fold increase in dermal testosterone concentrations, while sex hormones in females showed no change. The stress hormone cortisol was significantly higher in senescent than in reproductive individuals, independent of sex. Dermal corticosterone concentrations were highest in senescent females followed by senescent males, and lowest in reproductive individuals regardless of sex. Dermal swabs provide an effective and noninvasive means for evaluating octopus hormones. Application of these indicators may be imperative as cephalopods are more commonly cultured in captivity for experimentation, display, and consumption.

Determinants of maternal breast milk cortisol increase: Examining dispositional and situational factors

BackgroundBreast milk is a rich nutritional source, containing numerous proteins, carbohydrates, and hormones that impact long-term offspring development. Strikingly, predictors and correlates of breast milk composition remain largely unknown. Building on a previously discovered increase in breast milk cortisol concentration from 2 to 12 weeks postpartum, we investigated potential predictors of maternal breast milk cortisol in the first three months post-delivery by examining a suite of maternal dispositional (e.g., attachment, adverse childhood experiences or ACEs) and situational factors (e.g., partner support, self-efficacy). MethodsData from 73 mothers were collected prenatally, at birth, and 2-, 6- and 12 weeks postpartum. The analyses, which sought to predict postnatal changes in breast milk cortisol, included a pool of theoretically-sound constructs (Table 1) and an exploratory data-driven approach. We fit models differing in complexity as preregistered: 1) Random Forest models, capable of modeling interactions and non-linear relationships, and 2) Bayesian linear models, allowing to model change over time while less prone to overfitting. ResultsOverall, we found that no single variable had strong predictive value beyond the known predictors of cortisol, such as time since awakening and time of collection. However, results from both models suggest that ACEs carry information that warrants future investigations, pointing towards a negative relationship with cortisol concentration in breast milk, albeit with a minimal effect size. ConclusionUsing sophisticated models, we found that early life stress may play a role in physiological stress markers in breast milk in the first three months postpartum, with potential implications for offspring development.

Age-Related Difference in Cognitive Performance under Severe Whole-Body Hyperthermia Parallels Cortisol and Physical Strain Responses.

Background and Objectives: To date, understanding age-related changes in cognitive processes during heat exposure still needs to be better-understood. Thus, the main aim of the current study was to evaluate the effects of whole-body hyperthermia (WBH), i.e., a ≈ 2.5 °C increase in rectal temperature (Tre) from overnight-fast baseline value, on cognitive functioning in old and young men and to explore factors, such as stress and thermophysiological strain, that could influence such changes. Materials and Methods: Ten young (19-21 years of age) and nine old (61-80 years of age) healthy men underwent an experimental trial with passive lower-body heating in hot water immersion (HWI) at 43 °C (HWI-43 °C) until Tre reached 39 °C in old adults and 39.5 °C in young adults. Cognitive performance and cortisol concentration were assessed before and after HWI, and the physiological strain index (PSI) was assessed during HWI-43 °C. Results: PSI was lower and cortisol concentration was greater after HWI-43 °C in the old group compared with the young group (p < 0.05). Surprisingly, hyperthermia improved cognitive flexibility only in old adults, whereas short-term and visual recognition memories were maintained in both age groups. Conclusions: A ≈ 2.5 °C increase in rectal temperature can improve executive function in old adults, and this increase parallels the increased cortisol concentration and the lower thermophysiological strain under severe WBH conditions.

Changes in hair cortisol concentration in intrauterine device initiators: A prospective cohort study

ObjectivesPrior studies found increased hair cortisol concentration (a surrogate marker for hypothalamic-pituitary-adrenal axis activation) in users of the levonorgestrel intrauterine device (LNG 52 mg IUD). We evaluated change in hair cortisol and psychometric tests in women initiating a copper (CuT380 IUD) or LNG 52 mg IUD. Study designWe prospectively enrolled healthy women initiating an LNG 52 mg IUD or CuT380 IUD. Participants provided hair and blood samples and completed psychometric inventories (Patient Health Questionnaire-9, Positive and Negative Affect Schedule, and Psychological General Well-Being Index) after IUD insertion and at 6 and 12 months. We used liquid chromatography with tandem mass spectrometry to measure hair cortisol concentrations. We compared hair cortisol concentrations and psychometric test changes from baseline to 6 and 12 months using independent two-sample t tests. ResultsWe enrolled 39 of our targeted 86 participants (LNG 52 mg IUD 26, CuT380 IUD 13). Thirty-eight subjects (LNG 52 mg IUD 25, CuT380 IUD 13) completed 6 months of follow-up. We found no difference between cohorts in the mean change in hair cortisol concentrations at 6 months (LNG 52 mg IUD n = 21 [−0.01 pg/mg (95% CI −1.26, 1.23); CuT380 IUD n = 13 [−1.31 pg/mg (−3.36, 0.73)]). While psychometric inventory results remained within normal ranges, LNG 52 mg IUD users reported a trend toward more favorable changes over time. ConclusionsWe did not find clinically important differences in hair cortisol concentrations following initiation of a CuT380 IUD or LNG 52 mg IUD; psychometric inventories demonstrated no adverse effect of hormonal IUDs on mood. ImplicationsOur findings of similar hair cortisol concentrations following the initiation of either the LNG 52 mg IUD or CuT380 IUD suggest that hormonal IUDs do not increase cortisol concentrations or alter stress reactivity, and favorable effects on psychometric inventories provide further reassurance that the LNG 52 mg IUD has no adverse impact on mood. Trial registration numberClinicalTrials.gov NCT03499379

The effect of training load stress on salivary cortisol concentrations, health parameters and hematological parameters in horses

The performance of sport horses is conditioned not only by the quality of its gene pool, but also by a large number of external factors. The most dominant being nutrition, quality of breeding, level of zootechnical care and the quality of the sports rider and coach. Important factor is the process of individuals’ adaptation to the training load occurring during the training itself. This study was focused on the analysis of salivary cortisol levels as well as hematological and biochemical blood parameters in relation to load to which the tested horses were subjected. In the study 14 horses of sport breeds were analyzed a all tested horses were in the same (medium) level of training load. Tested horses underwent following stages of workload – transportation, jumping training, parkour competition, treadmill training, riding training, shoeing and lunging of various intensity. Saliva samples were obtained using a tampon on a string which was inserted into horse's oral cavity, chewed by the horse and placed in a sterile tube with a closable lid. Afterwards, the samples were then stored in deep-freezing boxes at temperature of −80 °C. The EIA cortisol kit was used in this study. The absorbance was read at the wavelength of 450 nm against a reference wavelength of 620–630 nm or a blank sample. Blood samples were obtained at the beginning of the experiment, after half a year of running the experiment and at the end of the experiment from v. jugularis. Hematological analysis were carried out using automatic hematologic analyser and multiple parameters were observed. Analysis of biochemical parameters in blood serum were realized using commercial DiaSys kits and semiautomatic biochemical spectrophotometer. Sodium, potassium and chlorides were measured using automatic analyzer EasyLytePlus. In all monitored forms of exercise (transportation, jumping training, parkour competition, treadmill training, riding training, horse shoeing, lunging), an increase in cortisol concentrations immediately after the exercise was recorded, but only spotted statistically significant differences were found during the transportation of monitored horses. The levels of blood parameters were within the reference range during the experiment period. From a comprehensive evaluation of the results, it can be stated that there were no visible health changes to the horses that underwent the experimental load and that manipulation with horses is an important factor that has effect on horses’ stress response. In general, the results of this study show no visible impact of training and/or load on the health status of horses over entire 12-month duration of the experiment.

Acute stress impairs visual path integration

Acute stress exerts substantial effects on episodic memory, which are often mediated by glucocorticoids, the end-product of the hypothalamic-pituitary-adrenal axis. Surprisingly little is known, however, about the influence of acute stress on human spatial navigation. One specific navigational strategy is path integration, which is linked to the medial entorhinal cortex, a region harboring glucocorticoid receptors and thus susceptible for stress effects. Here, we investigated effects of acute stress on path integration performance using a virtual homing task. We divided a sample of healthy young male participants into a stress group (nstress = 32) and a control group (ncontrol = 34). The stress group underwent the socially evaluated cold-pressor test, while the control group underwent a non-stressful control procedure. Stress induction was confirmed via physiological and subjective markers, including an increase of salivary cortisol concentrations. We applied linear mixed models to investigate the effect of acute stress on path integration depending on task difficulty and the presence or absence of spatial cues. These analyses revealed that stress impaired path integration especially in trials with high difficulty and led to greater decline of performance upon removal of spatial cues. Stress-induced deficits were strongly related to impaired distance estimation, and to a lesser extent to compromised rotation estimation. These behavioral findings are in accordance with the hypothesis that acute stress impairs path integration processes, potentially by affecting the entorhinal grid cell system. More generally, the current data suggests acute stress to impair cognitive functions mediated by medial temporal lobe regions outside the hippocampus.

Non-Invasive Biomarkers in Saliva and Eye Infrared Thermography to Assess the Stress Response of Calves during Transport.

Animal transport is currently a stressful procedure. Therefore, animal-based indicators are needed for reliable and non-invasive welfare assessment. Saliva is a biospecimen with potential validity for the determination of cortisol and oxidative stress, although its use to assess calf welfare during transport has never been tested. Similarly, the applicability and reliability of infrared thermography to assess temperature change during calves' transport have never been evaluated. These objectives were outlined following the known and growing need to identify non-invasive methodologies for stress assessment in bovines. This study was conducted on 20 calves of the Arouquesa autochthone breed, at about nine months of age, during their transport to slaughter. For each animal, saliva samples and thermographic images of the eye were collected at three time points: before transport, after transport, and at slaughter. The saliva was then processed to measure cortisol levels and oxidative stress parameters (reactive oxygen species, thiobarbituric acid reactive substance, carbonyls, and advanced oxidation protein products), and the images were analyzed using FLIR Tools+ software. There was an increase in cortisol concentration and oxidative stress parameters (reactive oxygen species, thiobarbituric acid reactive substance, carbonyls, and advanced oxidation protein products) in saliva after transport. An increase in eye temperature triggered by transport was also observed. The cortisol and eye temperature results at slaughter were returned to values similar to those before transport; however, the values of oxidative stress remained increased (mainly TBARS values). These non-invasive techniques seem to be reliable indicators of stress in bovine transport, and oxidative stress parameters in saliva may be a persistent marker for welfare assessment.

Assessment of the hypothalamic-pituitary-adrenocortical axis function using a vasopressin stimulation test in neonatal foals.

Bacterial sepsis is the leading cause of death in foals and is associated with hypothalamic-pituitary-adrenocortical axis (HPAA) dysfunction. HPAA function can be evaluated by an arginine-vasopressin (AVP) stimulation test. Administration of AVP will stimulate a dose-dependent rise in systemic adrenocorticotropin-releasing hormone (ACTH) and cortisol in neonatal foals. There will be no response seen in corticotropin-releasing hormone (CRH) and baseline AVP will be within reference interval. Twelve neonatal foals, <72 hours old. HPAA function was assessed in foals utilizing 3 doses of AVP (2.5, 5, and 7.5 IU), administered between 24 and 48 hours of age in this randomized cross-over study. Cortisol, ACTH, CRH and AVP were measured at 0 (baseline), 15, 30, 60 and 90 minutes after AVP administration with immunoassays. The fold increase in cortisol and ACTH was calculated at 15 and 30 minutes compared to baseline. All doses of AVP resulted in a significant increase in cortisol concentration over time, and a dose-dependent increase in ACTH concentration over time. ACTH and cortisol were significantly increased at 15 and 30 minutes, respectively after all 3 doses of AVP compared to baseline (P < .01). There was no change in endogenous CRH after stimulation with AVP. Administration of AVP is safe and results in a significant rise in ACTH and cortisol in neonatal foals. A stimulation test with AVP (5 IU) can be considered for HPAA assessment in septic foals.

Post-Traumatic Stress Response and Appendicitis in Children-Clinical Usefulness of Selected Biomarkers.

Acute appendicitis is an inflammatory process which is one of the most frequent global causes of surgical interventions in children. The goal of the study was to determine whether acute phase proteins, that is, C-reactive protein (CRP), procalcitonin (PCT) and neutrophil gelatinase-associated lipocalin (NGAL), interleukin 6 (IL-6), transforming growth factor-beta1 (TGF-β1) and cortisol (HC) play a role in the pathomechanism of post-trauma stress response of the organism and to establish the impact of the applied surgical procedure and/or of inflammation on their concentrations. An additional purpose was to establish the clinical usefulness of the studied biomarkers in the diagnostics of appendicitis. CRP concentrations were quantified via the immunoturbidimetric method, while the levels of IL-6 and PCT were assessed using a bead-based multiplexed immunoassay system in a microplate format (Luminex xMAP technology); NGAL, TGF-β1 and cortisol concentrations were determined via the enzyme-linked immunosorbent assay (ELISA) technique. All the investigated biomarkers were assayed twice, i.e., immediately before the surgery and 12-24 h after its completion. Significant increases in CRP, IL-6 and PCT concentrations were found in all children subjected to laparoscopic surgeries (p = 0.001, p = 0.006, and p = 0.009, respectively) and open (classic) surgeries (p = 0.001, p = 0.016, and p = 0.044, respectively) compared to the initial concentrations. The patients undergoing classical surgery moreover presented with significant (p = 0.002, and p = 0.022, respectively) increases in NGAL and TGF-β1 levels after the procedures. In a group of children undergoing laparoscopic surgery, the appendicitis induced an increase in cortisol concentration, whereas in patients undergoing classical surgery the increase in the levels of this biomarker was caused by the type of performed surgical procedure. Simultaneously assaying the levels of CRP, NGAL and IL-6 (p = 0.008, p = 0.022, and p = 0.000, respectively) may prove useful in clinical practice, enabling the diagnosis of appendicitis in paediatric patients reporting to a hospital with abdominal pains, in addition to data from anamnesis and from clinical or ultrasound examination. The performed study confirms the participation of examined biomarkers in the pathomechanism of post-injury stress reaction of the organism to surgical trauma.

Bio-Inspired Molecularly Imprinted Polymer Electrochemical Sensor for Cortisol Detection Based on O-Phenylenediamine Optimization.

This paper presents a comprehensive investigation of the various parameters involved in the fabrication of a molecularly imprinted polymer (MIP) sensor for the detection of cortisol. Parameters such as monomer concentration, electropolymerization cycles, pH, monomer-template ratio, template removal technique, and rebinding time were optimized to establish a more consistent and effective method for the fabrication of MIP sensors. Under the optimized conditions, the MIP sensor demonstrated a proportional decrease in differential pulse voltammetry peak currents with increasing cortisol concentration in the range of 0.1 to 100 nM. The sensor exhibited excellent sensitivity, with a limit of detection of 0.036 nM. Selectivity experiments using a non-imprinted polymer sensor confirmed the specific binding affinity of the MIP sensor for cortisol, distinguishing it from other steroid hormones. This study provides crucial insights into the development of a reliable and sensitive strategy for cortisol detection using O-PD-based MIPs. These findings laid the foundation for further advancements in MIP research.

Features of hormonal homeostasis in women with functional hypothalamic amenorrhea and premature ovarian insufficiengy caused by posttraumatic stress disorder.

Posttraumatic stress disorder can cause severe and prolonged amenorrhea in women. Since the beginning of the war in Ukraine, there has been an increased number of visits for the medical care of female military servicewomen and displaced women with this problem.The objective: to perform a comprehensive study of the characteristics of the hormonal levels of the hypothalamic-pituitary-ovarian axis and the stress hormone cortisol, as well as to analyze the presence of autoantibodies to ovarian tissue in women with secondary amenorrhea caused by posttraumatic stress disorder.Materials and methods. The levels of hypothalamic-pituitary-ovarian hormones (follicle-stimulating hormone (FSH), luteinizing hormone (LH), prolactin, estradiol, progesterone, free testosterone and anti-Mullerian hormone (AMH), as well as cortisol (urine), autoantibodies to ovarian tissue from 54 military servicewomen and displaced women with amenorrhea caused by posttraumatic stress disorder (main group). 35 (64.8%) women (the 1st subgroup) were diagnosed with functional hypothalamic amenorrhea (FHA), 19 (35.2%) patients (the 2nd subgroup) were diagnosed with premature ovarian failure (POF).The control group included 23 women of reproductive age (18–37 years), who were in a state of psychological comfort and had no menstrual cycle disorders.Results. As a result of the study, a significant (p&lt;0.05) decrease of all pituitary-ovarian hormones was found in women with FHA, except for free testosterone and AMH. In patients with POF, there was an increased concentration of pituitary hormones and a decreased amount of estradiol by 2.2 times, progesterone by 3.9 times, and AMH by 21.0 times compared to the control group.A 1.5-fold increase in prolactin level in patients with POF compared to the indicator of the control group is noteworthy. The presence of autoantibodies in the group with POF was detected in 26.3% of cases (in the group of women with FHA – only in 2.9% of cases; p&lt;0.05), which may be a manifestation of polyglandular autoimmune syndrome.In both examined subgroups of the main group, a probable increased cortisol concentration in daily urine was determined (by 2.0 times in the 1st subgroup and by 2.1 times in the 2nd subgroup compared to the control), which indicates a stressogenic dysfunction of the adrenal glands.Conclusions. Posttraumatic stress disorder can have an irreversible pathological effect on the reproductive system of women. In women with stressogenic functional hypothalamic amenorrhea there is a deep depression of the hypothalamic-pituitary-ovarian axis with preserving the follicular ovarian apparatus.In women with premature ovarian failure (POF) with long-term stress, there is a sharp depletion of the ovarian reserve. The presence of autoantibodies to ovarian tissue indicates the involvement of the immune system in the pathogenetic chain of development of amenorrhea, the severity of the lesion and the higher speed of the follicle atresia process by POF.