The study measured the hormonal and protein markers of acute stress, those of oxidative stress and total antioxidant capacity (TAC) in swine oral fluid, determined which of these parameters would be the most appropriate for future livestock welfare assessment and established the time when the samples should be taken. Stress was induced in 7 out of 14 castrated six-week-old Danbred×Duroc pigs by immobilisation on a nasal snare at 8 a.m., 1 p.m., and 6 p.m. and samples were taken both directly after the stressor was applied and 30 min later. The remaining pigs were the control group, which were not immobilised; their samples were taken at the same times. The concentrations of hormones and malondialdehyde (MDA) were measured using liquid chromatography with tandem mass spectrometry, while those of alpha-amylase and TAC were measured using spectrophotometry. The levels of cortisol and cortisone increased with statistical significance immediately after the acute stress response and 30 min later. A cut-off value set at 0.25 ng/mL cortisol concentration was capable of distinguishing between the stressed and control groups with 100% accuracy in evening samples and 95% accuracy overall. Prednisolone was not present, and the levels of testosterone and corticosterone were low and not distinctive. Alpha-amylase became significantly more concentrated during stress induction and 30 min later. The TAC and MDA levels rose after the stress but without statistical significance. The most suitable markers of acute stress were cortisol, cortisone and alpha-amylase. Oral fluid is a reliable material for monitoring the level of pigs’ stress and should be collected in the evening.

There is a balance between oxidative stress, antioxidant capacity and immune response. Their roles in physiological and behavioural mechanisms are important for the maintenance of the organism’s internal equilibrium. This study aimed to evaluate the antioxidant effects of the exogenous alga Spirulina platensis (Arthrospira platensis) in a stress-induced rat model, and to describe its possible mechanism of action. Thirty-six adult male Sprague Dawley rats were separated into four groups: control (C), stress (S), S. platensis (Sp), and S. platensis + stress (SpS). The rats in groups Sp and SpS were fed with 1,500 mg/kg b.w./day Spirulina platensis for 28 days. All rats were exposed to prolonged light phase conditions (18 h light : 6 h dark) for 14 days. The SpS and S groups were exposed to stress by being kept isolated and in a crowded environment. Blood samples were obtained by puncturing the heart on the 28th day. The effect of stress on serum corticosterone, oxidative stress markers (TOS, TAC, PON1, OSI) and immunological parameters (IL-2, IL-4, IFN-ɣ) were tested. Also, the brain, heart, intestines (duodenum, ileum, and colon), kidney, liver, spleen, and stomach of the rats were weighed. Serum corticosterone levels were higher in the S group than in the C group, and significantly lower in the SpS group than in the S group. Mean total antioxidant capacity were lower in the S group than in the C group, and Spirulina reversed this change. Although not significantly different, IL-2 was lower in the S group than in the C group. However, in the SpS group, IL-2 increased due to Spirulina platensis mitigating effects of stress. Male rats fed a diet with Spirulina platensis could experience significantly milder physiological changes during stress, although stress patterns may be different. Exogenous antioxidant supplements merit further investigation in animals and humans where the endogenous defence mechanism against stress may not be sufficient.

Objective—To determine whether trilostane or ketotrilostane is more potent in dogs and determine the trilostane and ketotrilostane concentrations that inhibit adrenal gland cortisol, corticosterone, and aldosterone secretion by 50%. Sample—24 adrenal glands from 18 mixed-breed dogs. Procedures—Adrenal gland tissues were sliced, placed in tissue culture, and stimulated with 100 pg of ACTH/mL alone or with 5 concentrations of trilostane or ketotrilostane. Trials were performed independently 4 times. In each trial, 6 samples (1 for each time point) were collected for each of the 5 concentrations of trilostane and ketotrilostane tested as well as a single negative control samples. At the end of 0, 1, 2, 3, 5, and 7 hours, tubes were harvested and media and tissue slices were assayed for cortisol, corticosterone, aldosterone, and potassium concentrations. Data were analyzed via pharmacodynamic modeling. One adrenal slice exposed to each concentration of trilostane or ketotrilostane was submitted for histologic examination to assess tissue viability. Results—Ketotrilostane was 4.9 and 2.4 times as potent in inhibiting cortisol and corticosterone secretion, respectively, as its parent compound trilostane. For trilostane and ketotrilostane, the concentrations that inhibited secretion of cortisol or corticosterone secretion by 50% were 480 and 98.4 ng/mL, respectively, and 95.0 and 39.6 ng/mL, respectively. Conclusions and Clinical Relevance—Ketotrilostane was more potent than trilostane with respect to inhibition of cortisol and corticosterone secretion. The data should be useful in developing future studies to evaluate in vivo serum concentrations of trilostane and ketotrilostane for efficacy in the treatment of hyperadrenocorticism.

Objective-To investigate the mechanisms by which corticosteroid administration may predispose cats to congestive heart failure (CHF). Animals-12 cats receiving methylprednisolone acetate (MPA) for the treatment of dermatologic disorders. Procedure-The study was conducted as a repeated-measures design. Various baseline variables were measured, after which MPA (5 mg/kg, IM) was administered. The same variables were then measured at 3 to 6 days and at 16 to 24 days after MPA administration. Evaluations included physical examination, systolic blood pressure measurement, hematologic analysis, serum biochemical analysis, thoracic radiography, echocardiography, and total body water and plasma volume determination. Results-MPA resulted in a substantial increase in serum glucose concentration at 3 to 6 days after administration. Concurrently, RBC count, Hct, and hemoglobin concentration as well as serum concentrations of the major extracellular electrolytes, sodium and chloride, decreased. Plasma volume increased by 13.4% (> 40% in 3 cats), whereas total body water and body weight slightly decreased. All variables returned to baseline by 16 to 24 days after MPA administration. Conclusions and Clinical Relevance-These data suggest that MPA administration in cats causes plasma volume expansion as a result of an intra- to extracellular fluid shift secondary to glucocorticoid-mediated extracellular hyperglycemia. This mechanism is analogous to the plasma volume expansion that accompanies uncontrolled diabetes mellitus in humans. Any cardiovascular disorders that impair the normal compensatory mechanisms for increased plasma volume may predispose cats to CHF following MPA administration.

Chickens in a low-stress environment (heterophil/lymphocyte ratio 0.31) were given feed containing 30, 40, or 60 mg of corticosterone/kg of feed for 0.5 hour. Between 0.5 to 12 hours later, chickens were exposed to Escherichia coli via the air sac route. For each dose of corticosterone, there was an untreated control group that was exposed to E coli via the air sac route. The prevalence of pericarditis was reduced from 78 to 7% between 2 and 4 hours after exposure. Resistance was associated with heterophil/lymphocyte (H/L) ratios greater than 1.04. Peak H/L ratios correlated positively with amount of corticosterone in the feed. In one experiment, chickens were inoculated IV with sheep erythrocytes at various times after consumption of feed containing corticosterone. Suppression of antibody responsiveness was most pronounced 4 hours later. Antibody responsiveness correlated positively with lymphocyte numbers. Histologic examination of air sacs was made following euthanasia at various times after E coli exposure. Lesions observed in control chickens included: edema at 0.5 hour, beginning of heterophil infiltration at 1 hour, increased edema and heterophil infiltration at 2 hours, and severe edema and heterophil infiltration at 4 hours. Lesions were not observed in chickens that had been given feed containing 40 mg of corticosterone/kg of feed.

The effect of in-house transport on plasma corticosterone concentration and blood lymphocyte populations of laboratory mice was investigated. Mice were transported within a research facility at 0900 hours in a pattern designed to simulate that commonly used by investigators prior to experimental manipulation. Plasma corticosterone concentration and WBC count were determined at 0.25, 2, 4, 8, 12, and 24 hours after transport. A significant (P less than 0.05) increase in plasma corticosterone concentration was seen in mice immediately after transport. The normal circadian rhythm of plasma corticosterone concentration was altered for the subsequent 24-hour period. Corresponding significant (P less than 0.05) decreases in total WBC numbers, lymphocyte count, and thymus gland weight were observed. The decrease in total blood lymphocyte numbers at 4 hours was reflected in B-and T-lymphocyte populations. The subsequent acute increase in plasma corticosterone concentration was associated with alterations in the cellular components of the immune system. Results of the study indicated that routine in-house transport of laboratory mice should be considered a stressful stimulus.

Plasma cortisol and corticosterone responses of 8 clinically normal adult ferrets to synthetic ACTH (cosyntropin) were evaluated. Cosyntropin was administered iv at 4 dosages (0.5, 1.0, 5.0, and 10 micrograms/kg of body weight) at 2- to 4-week intervals, with blood samples collected 60 and 120 minutes after injection. After completion of the studies, an additional ACTH stimulation test was performed by administering cosyntropin (1.0 micrograms/kg) IM. The baseline plasma cortisol concentrations from all studies ranged from 25.9 to 235 nmol/L (mean +/- SEM = 73.8 +/- 7.0 nmol/L), and plasma corticosterone values ranged from 1.7 to 47 nmol/L (mean +/- SEM = 8.3 +/- 1.1 nmol/L). After iv administration of cosyntropin, plasma concentrations of cortisol and corticosterone increased significantly (P < 0.05) and reached peak values at 60 minutes; however, there were no significant differences between plasma cortisol or corticosterone responses to the 4 dosages of cosyntropin. Intramuscular administration of 1.0 Kg of cosyntropin/kg induced increases in plasma cortisol and corticosterone concentrations that were similar to the responses induced by iv administration of cosyntropin. The mean molar ratio of cortisol to corticosterone, calculated from the resting plasma concentrations, was approximately 9:1, whereas the ACTH-stimulated cortisol to corticosterone ratio was approximately 4:1. Results of this study indicated that administration of cosyntropin to clinically normal ferrets, at dosages ranging from 0.5 to 10 micrograms/kg, increased plasma concentrations of cortisol and corticosterone. Although cosyntropin stimulates the adrenocortical secretion of cortisol and corticosterone, cortisol appears to be the predominate circulating glucocorticoid in ferrets.