Effects of dermal dexamethasone application on ACTH and both basal and ACTH-stimulated cortisol concentration in normal horses

There are no data available regarding the systemic (adverse) effects which might be induced by topical/dermal glucocorticoids (GCs) application in the horse. Besides their widespread use for the treatment of a variety of peripheral inflammatory disorders such as atopic dermatitis, eczemas or arthritis in the horse, their surreptitious application has become a concern in doping cases in competition/performance horses. Assessing both basal and ACTH‐stimulated plasma cortisol as well as basal ACTH concentrations following application of dexamethsone‐containing dermal ointment is necessary to determine influences on hypothalamus‐pituitary‐adrenal (HPA) axis. Ten clinically healthy adult standardbred horses (6 mares, 4 geldings) were rubbed twice daily each with 50 g dexamethasone‐containing ointment on a defined skin area (30 × 50 cm) for 10 days. RIA and chemiluminescent enzyme immuno‐metric assay were used to determine resting and ACTH‐stimulated plasma cortisol and basal ACTH concentrations, respectively. HPA feedback sensitivity and adrenal function were measured by a standard ACTH stimulation test. Dermal dexamethasone suppressed significantly the resting plasma cortisol level (to 75–98%) below baseline (P < 0.001) within the first 2 days and decreased further until day 10. ACTH stimulation test showed a markedly reduced rise in plasma cortisol concentrations (P < 0.001 vs. baseline). Plasma ACTH level decreased also during topical dexamethasone application. The number of total lymphocytes and eosinophil granulocytes was reduced, whereas the number of neutrophils increased. No significant change of serum biochemical parameters was noted. Dermal dexamethasone application has the potential to cause an almost complete and transient HPA axis suppression and altered leukocyte distribution in normal horses. The effects on HPA axis function should be considered in relation to the inability of animals to resist stress situations. The data further implicate that percutaneously absorbed dexamethasone (GCs) may cause systemic effects relevant to ‘doping’.     

Evaluation of plasma catecholamine and serum cortisol concentrations in horses with colic

OBJECTIVE: To evaluate plasma epinephrine and norepinephrine concentrations and serum cortisol concentration in horses with colic and assess the relationship of these variables with clinical signs, routinely measured clinicopathologic variables, and outcome in affected horses. DESIGN: Prospective observational study. ANIMALS: 35 horses with colic. PROCEDURE: Blood samples were collected within 30 minutes of arrival at the veterinary hospital from horses referred because of colic. Plasma and serum samples were analyzed for cortisol, epinephrine, norepinephrine, lactate, and electrolyte concentrations and acid-base variables. Heart rate at admission and outcome (survival or nonsurvival) were recorded. Univariate logistic regression was used to calculate crude (unadjusted) odds ratios and 95% confidence intervals. RESULTS: Of the 35 horses with colic, 26 survived. Higher plasma epinephrine, plasma lactate, and serum cortisol concentrations were significantly associated with increased risk of nonsurvival, but plasma norepinephrine concentration was not associated with outcome. Plasma epinephrine concentration was significantly correlated with heart rate (r = 0.68), plasma lactate concentration (r = 0.87), blood pH (r = -0.83), anion gap (r = 0.74), and base excess (r = -0.81). CONCLUSIONS AND CLINICAL RELEVANCE: The risk of death appears to be greater in colic-affected horses with high circulating concentrations of epinephrine and cortisol. The correlation of epinephrine with other biochemical markers of illness severity and with heart rate indicates that the degree of sympathetic activation in horses with colic can be inferred from routinely measured variables.     

Salivary and plasma cortisol as an index of stress in goats

Total assayable cortisol in plasma was highly correlated (r = 0.97) with physiologically active free cortisol in plasma after routine management procedures in 1- to 3-weeks-old goats. Transport of adult goats caused significant increases (P less than 0.001) in free cortisol in saliva and in free and total cortisol in plasma. No difference (P greater than 0.05) between concentrations of free cortisol in saliva and in plasma was apparent before or after transport. The results demonstrated that the salivary cortisol method is a useful measure of stress in adult goats, and that the relationship between free and total cortisol in plasma, and the adrenocortical response to transport, appear to be similar in sheep and goats.     

Urinary cortisol:creatinine ratios in healthy horses and horses with hyperadrenocorticism and non-adrenal disease

Urinary cortisol and creatinine concentrations, and the cortisol:creatinine ratio were compared between 12 healthy horses (group 1), 13 horses with Cushing's disease (group 2), and eight horses with dysautonomia syndrome (equine grass sickness) (group 3). The mean (sd) urinary cortisol concentrations were 112 (55.7), 250 (357) and 864 (526) nmol/litre in groups 1, 2 and 3, respectively; the mean (sd) urinary creatinine concentrations were 18.9 (7.3), 12.0 (6.7) and 45.2 (26.4) nmol/litre in groups 1, 2 and 3, respectively, and the mean (sd) ratios were 6.1 (2.6), 19.8 (23.8) and 21.3 (14.5) (x 10(-6)) in groups 1, 2 and 3, respectively. The urinary cortisol and creatinine concentrations were significantly greater in group 3 than in groups 1 and 2, but the ratios were not significantly different, although there was a trend (P=0.076) towards higher values in groups 2 and 3. A diagnostic cut-off in the cortisol:creatinine ratio for the confirmation of Cushing's disease of more than 6.9 x 10(-6) was associated with a diagnostic sensitivity and specificity of 92.3 and 75.0 per cent, respectively, when compared with healthy horses. However, when group 3 horses were included, a cut-off of more than 7.4 x 10(-6) was associated with a diagnostic sensitivity and specificity of 84.6 and 54.5 per cent, respectively.     

Clearance of bromosulphthalein from plasma as a measure of hepatic function in normal horses and in horses with liver disease

Single intravenous injections of bromosulphthalein (BSP) were given to horses and the change in plasma concentration of BSP with time was analysed by computer to obtain the proportionality transfer constants 'a', 'h' and 'b'. No age, weight or sex differences in BSP clearance were found in normal horses. The technique was non-invasive, repeatable and suitable for conscious animals. The measurement of the transfer constants 'a', 'h' and 'b', helped to provide an accurate guide to diagnosis and prognosis of liver disease.     

Aldosterone plasma concentration in horses with heart valve insufficiencies

In humans and small animals, heart disease can lead to an increase in aldosterone, and the aldosterone level correlates with the severity of the heart disease. In horses similar interactions may be possible and may lead to an increase in aldosterone in horses with heart valve insufficiencies. In a prospective clinical trial eight healthy horses (control group) and 40 horses with heart valve disease were examined. In all horses, a clinical (auscultation), electro- and echocardiographic examination was performed and aldosterone plasma concentration was determined. The median aldosterone plasma concentration in the control group was 23.95 pg/ml. Twenty-one out of 40 horses with heart valve insufficiencies and without dimensional changes by echocardiography (group 1) showed a median aldosterone plasma concentration of 45.5 pg/ml. Five out of the 40 horses had a left atrial (LA) dilation and an average LA size with 147.6+/-11 mm (group 2) and a median aldosterone plasma concentration of 95.9 pg/ml. Five other horses had a left ventricular (LV) dilation with an average LV size of 141.6+/-6.8 mm (group 3) and a median aldosterone plasma concentration of about 115.3 pg/ml. In this group a positive correlation between aldosterone plasma concentration and LV existed (r=0.9, P=0.03). Nine horses with both LA (152.8+/-11.4 mm) and LV dilatation (145+/-9 mm, group 4) had a median aldosterone plasma concentration of 161.2 pg/ml. Significant differences of the aldosterone concentrations were observed between the control group and the horses with LA and LV dilation (group 4, P=0.0005), as well as between group 1 (horses with heart valve insufficiency but without dilation) and group 4 (P=0.0006). The study confirms that, as reported for other species, aldosterone rises as the severity of valvular disease increases. However, in this study, as there is only significant difference from normal in the most affected group, it will require further study before the plasma aldosterone level can be relied on as an indicator of the severity of heart disease in an individual horse.     

Salivary cortisol in pigs following adrenocorticotrophic hormone stimulation: comparison with plasma levels

Four prepubertal pigs were prepared with venous catheters and housed in metabolism cages. Plasma and saliva samples were taken at 15-min intervals over a 105-min period and analysed by radioimmunoassay for total (i.e. free and bound) cortisol content. Adrenocorticotrophic hormone (ACTH) was given i.v. at three different doses (0.5, 1.0 and 2.0 mg) after the second sample and the cortisol responses were compared with pretreatment values and levels observed after saline vehicle administration. Basal levels of salivary cortisol were approximately 10% of those in plasma. ACTH induced significant increases in plasma and salivary cortisol but in no case was a dose/response relationship detected. Plasma cortisol showed a maximum increase of approximately 230% whereas salivary cortisol increased only by about 130%, indicating that salivary cortisol is a less sensitive indicator of adrenal activity than plasma cortisol in this species. Estimation of salivary cortisol concentrations may offer practical advantages for the assessment of stress responses in intensively housed pigs.     

Perioperative plasma cortisol concentration in the horse

The cortisol response to anaesthesia and surgery was studied in 2 groups of horses undergoing either abdominal or non-abdominal surgery. The preoperative mean plasma cortisol concentration (pcc) of 381.7 nmol/l (s.d. 254.7) was markedly higher in the abdominal group than the early-morning mean pcc of 115.6 nmol/l (s.d. 78.4) in the non-abdominal group. During halothane anaesthesia and surgery the mean pcc increased significantly (p < 0.05) from the preoperative mean of 119.2 to 215.9 nmol/l (s.d. 79.8) after 30 min of surgery in the non-abdominal group. In the abdominal group a decrease occurred after induction of anaesthesia and surgical preparation, but increased during surgery to a mean pcc of 418.1 nmol/l (s.d. 236.5). In the postoperative period a large decrease in the mean pcc occurred after 24 h in the abdominal group. It was only after 60 h that the pcc (153.2 nmol/l) equalled the pcc of the non-abdominal group (171.4 nmol/l) at 24 h. The slow decline over 60 h could be an indication of the prolonged recovery associated with abdominal surgery in the horse.     

Occlusal angles of cheek teeth in normal horses and horses with dental disease

The angle between the occlusal surface of the tooth and the horizontal plane of 687 cheek teeth from the skulls of 22 horses without gross dental disorders and 11 horses with dental disorders were measured by using stiff malleable wire as an imprint. Each measurement was repeated five times and the mean angle was recorded. In the normal skulls, the mean occlusal angles of the mandibular cheek teeth ranged from 19.2 degrees at the Triadan 06 position to 30 degrees at the 11 position, and these angles were significantly greater than the occlusal angles of the opposing maxillary cheek teeth (range 12.5 degrees to 18 degrees) at all the positions except the 06. The rostral mandibular cheek teeth had significantly lower occlusal angles than the caudal mandibular cheek teeth, but the converse was true for the maxillary teeth. In the skulls with dental disorders the occlusal angles of the mandibular cheek teeth ranged from 15.6 degrees to 28.5 degrees , and of the maxillary cheek teeth from 9.2 degrees to 16.4 degrees. They were not significantly different from the angles of the teeth from the normal skulls, except at the 06 position, where they were smaller.     

Histamine inhalation challenge in normal horses and in horses with small airway disease

A histamine inhalation challenge (HIC) procedure was developed to assess hyperreactive states in horses. Following clinical evaluation, percutaneous lung biopsies were performed on nine light breed mares aged 6 to 15 years. Five horses, with normal small airways, were classified as group A and four subjects with small airway disease (SAD) lesions formed group B. Pulmonary mechanics parameters were monitored following an aerosol of 0.9% saline and every 5 min for up to 30 min after HIC with 0.5% w/v of histamine diphosphate, administered through a face mask for 2.5 min. Tidal volume (VT) and airflow (V) values were obtained with a pneumotachograph. Transpulmonary pressure (delta Ppl) was measured by the esophageal balloon catheter method. Dynamic compliance (Cdyn), total pulmonary resistance (RL), end expiratory work of breathing (EEW) and respiratory rate (f) were calculated by a pulmonary mechanics computer. Group A horses had increases in RL, and decreases in Cdyn whereas horses in group B were hyperreactive and showed greater changes in EEW, Cdyn, and delta Ppl but with a relatively lower variation of RL. One horse in clinical remission from SAD, but with a high biopsy score (group B), and one clinically normal horse belonging to group A showed marked hyperreactivity as shown by increases in EEW, maximum change in delta Ppl and RL and decreases in Cdyn. These results suggest that the HIC described can be used as a method to investigate airway hyperreactivity and SAD in horses.     

Serum insulin concentrations in horses with equine Cushing's syndrome: response to a cortisol inhibitor and prognostic value

REASONS FOR PERFORMING STUDY: Serum insulin concentration and its use as a prognostic indicator in horses with equine Cushing's syndrome (ECS) have been poorly documented. OBJECTIVES: To examine daily insulin variations in horses with ECS and the effect of treatment using trilostane, a competitive inhibitor of 3beta-hydroxysteroid dehydrogenase. Further, we aimed to examine the relationship between baseline serum insulin concentration and survival in horses with ECS. METHODS: Basal serum insulin concentrations were measured in 20 confirmed ECS cases by taking blood at regular 4 h intervals for 24 h (1200, 1600, 2000, 2400, 0400 and 0800 h) before treatment (Day 0) and 10 days, and 30 days and 1-2 years after the onset of trilostane therapy. The temporal pattern of insulin was analysed using a linear mixed model approach, and the prognostic value of measurements on Day 0 assessed using receiver-operating characteristic analysis. RESULTS: Horses with ECS showed a diurnal pattern of serum insulin concentration, highest value at 1200 h, and this pattern was not altered by treatment with trilostane. Furthermore, despite a mild increase of serum insulin concentrations after 10 days of trilostane therapy, insulin concentration was unaffected in the long-term. Low serum insulin concentrations at the beginning of the trial were significantly associated with improved survival to 1-2 years. The 1200 h sampling before treatment had the highest prognostic value for prediction of survival with a sensitivity and specificity of at least 90% for serum insulin at < 62 and > 188 microu/ml to predict survival and nonsurvival, respectively. CONCLUSIONS AND POTENTIAL RELEVANCE: Insulin is a useful prognostic indicator for ECS, but potentially large variations can occur throughout a 24 h period, indicating a single sample may not be representative. Serum insulin concentration did not increase over 1-2 years in horses receiving trilostane therapy.     

Pulmonary artery pressures in normal horses and in horses affected with chronic obstructive pulmonary disease

Horses clinically affected with chronic obstructive pulmonary disease (COPD) were found to have pulmonary artery hypertension which was associated with systemic arterial hypoxia. The pulmonary hypertension in symptomatic COPD-affected horses was partially reversible upon remission of clinical signs or by oxygen administration. The induction of acute hypoxaemia caused an increase in pulmonary artery pressure in both normal and COPD-affected horses.     

Association of admission plasma D-dimer concentration with diagnosis and outcome in horses with colic

Background: Coagulopathies detected in horses with gastrointestinal problems seem to be associated with poor outcome. Plasma D‐Dimer concentration is a sensitive test for assessing coagulopathies. Hypothesis: Plasma D‐Dimer concentration tested on admission is related to diagnosis and outcome in horses with colic. Animals: Four hundred and ninety three horses referred for evaluation of abdominal pain. Methods: Prospective observational clinical study. Horses were grouped according to diagnosis (medical and surgical intestinal obstructions, ischemic disorders with and without intestinal resection, enteritis, peritonitis), outcome (survivors, nonsurvivors), and number of coagulopathies (normal profile, 1 or 2 coagulopathies, subclinical disseminated intravascular coagulation [DIC]). Blood samples were collected on admission and plasma D‐Dimer concentration, clotting times (PT and aPTT), and antithrombin activity were determined. Positive likelihood ratios (LR+) were calculated for evaluation of D‐Dimer cut‐off values, which were later tested in a logistic regression model. Results: Horses with enteritis or peritonitis had significantly (P < .001) higher plasma D‐Dimer concentrations and more severe coagulopathies on admission than horses with other diagnoses. Nonsurvivors also had significantly (P < .001) higher plasma D‐Dimer concentrations at presentation than did survivors, and those horses with subclinical DIC on presentation had an odds ratio (OR) 8.6 (95% confidence interval [CI], 3.3–22.5, P < .001) for nonsurvival. Finally, D‐Dimer concentrations >4,000 ng/mL had a LR+ of 5.9 and an OR 8.8 (95% CI, 4.5–17.1, P < .001) for nonsurvival. Conclusion and Clinical Importance: Plasma D‐Dimer concentration measured on admission can be used to facilitate diagnosis and outcome prediction in horses with colic. A potential cut‐off value for nonsurvival was found at approximately 4,000 ng/mL.     

Biochemical indices of vascular function, glucose metabolism and oxidative stress in horses with equine Cushing's disease

REASONS FOR PERFORMING STUDY: The mechanisms underlying the increased risk of laminitis in horses with equine Cushing's disease (ECD) are poorly understood. HYPOTHESIS: That abnormalities in glucose homeostasis, similar to those which cause microvascular dysfunction in human diabetics, contribute to development of laminitis in horses with ECD. METHODS: Thirty-one aged horses were divided into 3 groups based on clinical signs and dexamethasone suppression testing (DST). Group 1 (n = 12) had clinical ECD as evidenced by hirsutism. Group 2 (n = 10) had a positive DST but no hirsutism. Group 3 (n = 9) were controls without ECD, with a negative DST and no clinical evidence of ECD. Biochemical indices of glucose metabolism, vascular function and oxidative stress were determined in single morning blood samples. RESULTS: Group 1 had abnormalities in glucose homeostasis, including increased levels of glucose and insulin, compared to Groups 2 and 3. Groups 1 and 2 had significantly lower plasma thiol (PSH) levels and nonsignificantly lower albumin-corrected PSH levels than Group 3, consistent with oxidative stress. CONCLUSIONS AND POTENTIAL RELEVANCE: The observed abnormalities in glucose metabolism and oxidative stress could potentially contribute to development of laminitis in horses with ECD, by similar mechanisms to those that cause microvascular dysfunction in human diabetics. The absence of inter-group differences in the biochemical indices of vascular function precludes their use as preclinical diagnostic indicators of vascular dysfunction. The study also highlighted limitations in the premortem diagnosis of ECD.     

Myeloperoxidase assay in plasma and peritoneal fluid of horses with gastrointestinal disease

Gastrointestinal disorders, especially strangulating intestinal obstructions, are still a major cause of illness and death in the horse. Circulating lipopolysaccharides may activate both neutrophils and monocytes. The activated neutrophils release myeloperoxidase (MPO), a specific enzyme with strong oxidative activity. The aim of this study was to evaluate MPO concentrations in the plasma and peritoneal fluid (PF) of horses with colic and to check the hypothesis that these concentrations would be higher in a case of strangulating obstruction than in cases of nonstrangulating disease. By using a specific enzyme-linked immunosorbent assay for equine MPO, we determined the MPO concentrations in horses admitted to a clinic for colic. Horses with nonstrangulating or strangulating obstruction of the large intestine (NSLI or SLI), strangulating obstruction of the small intestine (SSI), or inflammatory bowel disease (IBD) were compared with healthy horses. The horses with SLI, SSI, or IBD had significantly higher MPO levels in plasma and PF than did those in the other 2 groups. The mean plasma level was significantly higher in the horses with NSLI than in the healthy horses. High MPO values in PF indicated necrotic bowel. These results show that neutrophil activation occurs during nonstrangulating and strangulating intestinal obstruction in horses and that the plasma and PF MPO concentrations may be a marker of the severity of the disease.