Libido, hormone concentrations in blood plasma and semen characteristics in Holstein bulls

Relationships among bull libido, serum hormone concentrations and semen characteristics were studied using 18 Holstein bulls that were 4 to 5 yr old. The hormones studied included testosterone, estradiol (E2), prolactin (PRL), LH and cortisol. Two ejaculates were collected three times per week from each bull during a 5-wk trial. During the last week of the trial, on a day semen was not collected, blood was collected from indwelling catheters every 15 min for 6 h to determine the hormonal profiles of each bull. On the following day, blood was sampled every 10 min before and after the time of semen collection. Libido factors were quantified, and semen volumes and sperm concentrations were recorded. The libido factors included reaction time to first service, latency time between the first and second semen collections, and duration of time the bull mounted the teaser prior to the first (TM1) and second (TM2) semen collection. Average reaction and latency times were correlated (r = .524; P = .026), as were TM1 and TM2 (r = .597; P = .015). Latency times were correlated with average TM2 (r = .669; P = .003). Average PRL concentrations were correlated with average latency times (r = .467; P = .05). Low libido bulls tended to have higher E2:testosterone ratios than did high libido bulls. Both PRL and cortisol concentrations increased at semen collection.     

Effect of hypothyroidism on blood lipid concentrations in horses.

OBJECTIVE: To measure and compare concentrations of selected blood lipids before and after thyroidectomy in horses. ANIMALS: 5 healthy adult mares. PROCEDURE: Mares were confirmed to be euthyroid. Thyroidectomy was performed, and hypothyroidism was confirmed. Selected blood lipid variables were measured before hypothyroidism was induced and weekly for 4 weeks after induction. Plasma concentrations of very low-density lipoprotein (VLDL), low-density lipoprotein (LDL), serum triglyceride (TG), total cholesterol (TC), and nonesterified fatty acid (NEFA) were measured. The composition of VLDL and LDL also was examined. RESULTS: Mean plasma concentrations of VLDL and LDL increased significantly after thyroidectomy. By 4 weeks after thyroidectomy, a ninefold increase in mean plasma concentration of VLDL and a threefold increase in LDL, compared with baseline values, were detected. After thyroidectomy, mean percentage of TG in VLDL increased significantly, whereas free cholesterol and cholesterol ester content decreased. Mean percentage of TG in LDL also increased by 3 to 4 weeks after thyroidectomy. Serum concentrations of TG and TC increased, whereas serum NEFA concentration decreased. CONCLUSIONS: Hypothyroidism significantly alters blood lipid concentrations of horses. After thyroidectomy, markedly high VLDL concentration, appearance of TG-rich VLDL, increased serum concentrations of TG and TC, and decreased blood concentration of NEFA were evident. CLINICAL RELEVANCE: Examination of blood lipid concentrations of horses may be useful for detecting naturally acquired hypothyroidism.     

Effects of a supplement containing chromium and magnesium on morphometric measurements, resting glucose, insulin concentrations and insulin sensitivity in laminitic obese horses

Reasons for performing study: Obesity and insulin resistance are risk factors for laminitis in equids and supplements containing chromium and magnesium might improve insulin sensitivity. Hypothesis: A supplement containing chromium, magnesium and other nutraceuticals would alter morphometric measurements, blood variables, and insulin sensitivity in laminitic obese horses. Methods: Twelve previously laminitic obese (body condition score ≥ 7/9) horses were randomly allocated to treatment (n = 6) and control (n = 6) groups and 2 obese horses with clinical laminitis were included in the treatment group. Treated animals received 56 g supplement with 0.25 kg oats once daily for 16 weeks. The supplement contained chromium (5 mg/day as yeast), magnesium (8.8 g/day as oxide/proteinate), and other nutraceuticals. Insulin‐modified frequently sampled i.v. glucose tolerance tests were performed with hay provided at 0, 8 and 16 weeks, and insulin sensitivity was estimated by minimal model analysis. Physical measurements were collected at the same points. Horses were not exercised. Results: Hyperinsulinaemia (>30 µu/ml) was detected in 12 of 14 horses prior to treatment. Glucose and insulin data from one mare with clinical laminitis were excluded because of persistent pain. Mean ± s.d. insulin sensitivity was 0.64 ± 0.62 × 10^?4 l/min/mu prior to treatment for the remaining 13 horses. Time and treatment × time effects were not significant for any of the variables examined, with the exception of resting insulin concentrations, which significantly increased over time (P = 0.018). Health status remained the same. Conclusions: The supplement containing chromium and magnesium evaluated in this study did not alter morphometric measurements, blood variables, resting insulin concentrations or insulin sensitivity in laminitic obese horses. Potential relevance: Additional research is required to determine the appropriate use of chromium and magnesium supplements in horses.     

Measurement of C‐peptide concentrations and responses to somatostatin,glucose infusion,and insulin resistance in horses

Reasons for performing study: Hyperinsulinaemia is detected in horses with insulin resistance (IR) and has previously been attributed to increased pancreatic insulin secretion. Connecting peptide (C‐peptide) can be measured to assess pancreatic function because it is secreted in equimolar amounts with insulin and does not undergo hepatic clearance. Hypothesis: A human double antibody radioimmunoassay (RIA) detects C‐peptide in equine serum and concentrations would reflect responses to different stimuli and conditions. Methods: A validation procedure was performed to assess the RIA. Six mature mares were selected and somatostatin administered i.v. as a primed continuous rate infusion, followed by 50 nmol human C‐peptide i.v. Insulin and C‐peptide concentrations were measured in horses (n = 6) undergoing an insulin‐modified frequently sampled i.v. glucose tolerance test, and in horses with insulin resistance (n = 10) or normal insulin sensitivity (n = 20). Results: A human RIA was validated for use with equine sera. Endogenous C‐peptide secretion was suppressed by somatostatin and median (range) clearance rate was 0.83 (0.15–1.61) ml/min/kg bwt. Mean ± s.d. C‐peptide‐to‐insulin ratio significantly (P = 0.004) decreased during the glucose tolerance test from 3.60 ± 1.95 prior to infusion to 1.03 ± 0.18 during the first 20 min following dextrose administration. Median C‐peptide and insulin concentrations were 1.5‐ and 9.5‐fold higher, respectively in horses with IR, compared with healthy horses. Conclusions: Endogenous C‐peptide secretion decreases in response to somatostatin and increases after dextrose infusion. Results suggest that relative insulin clearance decreases as pancreatic secretion increases in response to dextrose infusion. Hyperinsulinaemia in insulin resistant horses may be associated with both increased insulin secretion and decreased insulin clearance. Potential relevance: Both C‐peptide and insulin concentrations should be measured to assess pancreatic secretion and insulin clearance in horses.     

Increased plasma fructosamine concentrations in laminitic horses

Reasons for performing study: The use of plasma fructosamine concentration ([fructosamine]) as a marker of abnormal glucose homeostasis in laminitic horses has not been investigated. Hypothesis: Plasma fructosamine concentration may be higher amongst laminitic horses than normal horses; this might relate to underlying insulin resistance. Objectives: 1) To compare [fructosamine] between laminitic and normal horses. 2) To investigate associations between [fructosamine] at presentation in laminitic horses with a) single sample markers of insulin resistance and b) outcome. Methods: Plasma fructosamine concentration, fasting serum insulin concentration (insulin) and fasting plasma glucose concentration (glucose) were measured in 30 horses that presented with laminitis. Clinical details and follow‐up data were recorded. Plasma fructosamine concentration was also measured in 19 nonlaminitic control horses. Results: Laminitic horses had significantly higher mean [fructosamine] than normal horses (P<0.001). Thirteen of 30 laminitic horses had fasting hyperinsulinaemia, 2/30 had fasting hyperglycaemia. Statistically significant univariable correlations were identified between [fructosamine] and [glucose], [insulin] and the proxies RISQI and MIRG. Trends for association between [fructosamine] and negative outcome did not reach statistical significance. Conclusions and potential relevance: Increased mean [fructosamine] in laminitic horses may represent abnormal glycaemic control and [fructosamine] may become a clinically useful marker.     

Comparison between blood serum and salivary cortisol concentrations in horses using an adrenocorticotropic hormone challenge

Reasons for performing study: In horses, serum cortisol concentration is considered to provide an indirect measurement of stress. However, it includes both free and bound fractions. The sampling method is also invasive and often stressful. This is not the case for salivary cortisol, which is collected using a more welfare‐friendly method and represents a part of the free cortisol fraction, which is the biologically active form. Objectives: To compare salivary and serum cortisol assays in horses, in a wide range of concentrations, using an adrenocorticotropic hormone (ACTH) stimulation test, in order to validate salivary cortisol for stress assessment in horse. Methods: In 5 horses, blood samples were drawn using an i.v. catheter. Saliva samples were taken using swabs. Cortisol was assayed by radioimmunoassay. All data were treated with a regression method, which pools and analyses data from multiple subjects for linear analysis. Results: Mean ± s.d. cortisol concentrations measured at rest were 188.81 ± 51.46 nmol/l in serum and 1.19 ± 0.54 nmol/l in saliva. They started increasing immediately after ACTH injection and peaks were reached after 96 ± 16.7 min in serum (356.98 ± 55.29 nmol/l) and after 124 ± 8.9 min in saliva (21.79 ± 7.74 nmol/l, P<0.05). Discharge percentages were also different (225% in serum and 2150% in saliva, P<0.05). Correlation between serum and salivary cortisol concentrations showed an adjusted r²= 0.80 (P<0.001). The strong link between serum and salivary cortisol concentrations was also estimated by a regression analysis. Conclusions: The reliability of both RIAs and regression found between serum and salivary cortisol concentrations permits the validation of saliva‐sampling as a noninvasive technique for cortisol level assessment in horses.     

Serum thyroid hormone concentrations in New Zealand horses

Total thyroxine and total tri-iodothyronine concentrations were measured in the sera from 125 horses of mixed age, breed and sex, and varied clinical histories. While low serum thyroxine concentrations were detected in 35 horses, the majority of those horses had serum thyroxine values within the reference range when retested. Only one horse had a mildly decreased serum tri-iodothyronine concentration. Those horses in which the serum thyroxine concentration was low when retested had a normal thyrotropin releasing hormone stimulation test. Hypothyroidism was not diagnosed in any horses in this study. The low serum thyroxine concentrations measured in the present study were attributed to either normal fluctuations in serum concentrations in healthy horses, the effect of drugs, or to the effects of non-thyroidal illness. Because thyroid hormone concentrations are altered by many factors, hypothyroidism should not be diagnosed on the basis of a single low value and further testing, preferably including active stimulation of the thyroid gland, should be carried out.     

Cortisol concentrations in blood and urine of horses

A survey of the concentrations of cortisol in blood and urine samples taken from thoroughbred and standardbred horses after racing is presented. Statistical analysis showed the only significant difference between thoroughbred and standardbred horses was a higher cortisol concentration in thoroughbred urine. Urine volume and pH had no significant influence on the urinary cortisol concentration, however 9.5% of the urinary cortisol variation could be explained due to the influence of plasma cortisol concentration. The results of cortisol and ACTH administrations are also shown and compared with the survey results.     

Circannual variation in plasma adrenocorticotropic hormone concentrations in the UK in normal horses and ponies, and those with pituitary pars intermedia dysfunction

Reasons for performing study: Pituitary pars intermedia dysfunction (PPID) is a common endocrinopathy, frequently diagnosed via plasma adrenocorticotropic hormone (ACTH) concentrations. Seasonal variation in plasma ACTH concentrations has been described in normal horses prompting caution in diagnosing PPID at certain times of the year. The aims of this study were to determine appropriate reference intervals for equine plasma ACTH throughout the year; and to examine the circannual variation of plasma ACTH concentrations in PPID cases. Hypothesis: Plasma ACTH can be used as a test for PPID throughout the year with the use of appropriate reference intervals. Methods: Data for reference interval calculations were obtained from samples collected from inpatients of Liphook Equine Hospital (non‐PPID group, n = 156). Data from PPID cases (n = 941) were obtained from samples submitted to the Liphook Equine Hospital Laboratory from horses with a clinical suspicion of PPID found to have plasma ACTH concentrations greater than our upper reference interval for that time of year. Results: Upper limits for reference interval of plasma ACTH were 29 pg/ml between November and July and 47 pg/ml between August and October. Circannual variation in plasma ACTH occurred in both non‐PPID and PPID horses with the highest ACTH concentrations found between August and October in both groups (P<0.0001). The greatest difference between the 2 populations also occurred between August and October. Conclusions: Plasma ACTH can be used for the diagnosis and monitoring of PPID throughout the year with the use of appropriate reference intervals. These findings demonstrate an increase in pituitary gland secretory activity during the late summer and autumn in both normal and PPID cases.     

Effects of hypothyroidism and withholding of feed on plasma lipid concentrations,concentration and composition of very-low-density lipoprotein,and plasma lipase activity in horses

OBJECTIVE: To evaluate selected concentrations of blood lipids and lipase activities in euthyroid and hypothyroid horses deprived of feed for 96 hours. ANIMALS: 4 healthy adult mares and 4 thyroidectomized adult mares. PROCEDURE: Horses were deprived of feed for 96 hours. Blood samples were collected at 24-hour intervals and analyzed to determine concentrations of non-esterified fatty acid (NEFA), triglyceride (TG), total cholesterol (TC), and very-low-density lipoprotein (VLDL) as well as composition of VLDL. Plasma lipase activities were measured after feed was withheld for 96 hours and 12 days after resumption of feeding. RESULTS: Time significantly affected plasma NEFA, VLDL, TG, and TC concentrations in both groups of horses. During the 96-hour period, mean plasma concentrations of NEFA and VLDL increased 10-fold in euthyroid horses and increased 5-fold and 9-fold, respectively, in hypothyroid horses. Mean plasma TG concentrations increased 8-fold in both groups, and plasma TC concentrations significantly increased by 33 and 30%, respectively. Composition of VLDL was significantly affected by feed deprivation in euthyroid horses. Activities of lipoprotein lipase and hepatic lipase were significantly higher in feed-deprived horses. Activity of hepatic lipase was significantly lower in hypothyroid horses than in euthyroid horses. CONCLUSIONS AND CLINICAL RELEVANCE: Hypothyroidism did not significantly alter the magnitude of the response of blood lipids to feed deprivation. Thyroid hormones may reduce variability in blood lipid concentrations but do not determine susceptibility to hyperlipemia. Hypothyroidism does not appear to be a factor in the pathogenesis of hyperlipemia in horses.     

Hypertriglyceridemia with increased plasma insulin concentrations in cats

Metabolite, insulin and adiponectin concentrations and LDH, AST and ALT activities were measured in plasma of 142 client-owned cats (1–13 years old, 16 breeds) to set up a new criterion of hypertriglyceridemia (hyper-TG) with increased plasma insulin concentrations for early diagnosis of lipid metabolism abnormality including obesity. 25 cats with over 165 mg/dl of plasma triglyceride (TG) concentrations were decided as hyper-TG with increased plasma insulin concentrations, and prevalence of hyper-TG was 16.7% in young (1–6 years old) and 18.3% in old (>7 years old) cats examined. In the hyper-TG cats, their plasma TG concentrations increased to 6.6–7.4-fold of the values of control cats with 35–50 mg/dl of plasma TG and their plasma cholesterol, FFA and insulin concentrations and LDH and ALT activities increased significantly, whereas their plasma adiponectin concentrations decreased significantly compared to those in the control cats. Hyper-TG cats with significantly increased body weights and plasma insulin and decreased plasma adiponectin seemed to be in early stage of obesity accompanying increased plasma insulin concentrations. Increased TG, insulin, LDH and ALT and decreased adiponectin values in plasma seemed to be key factors for diagnosis of lipid metabolism abnormality at early stage in cats.     

Growth, nutrient utilization, radiographic bone characteristics and postprandial thyroid hormone concentrations in weanling horses fed added dietary fat

Thirty, 19-week-old Quarter Horses were utilized in a 112-d study to compare growth, nutrient utilization and post prandial thyroid hormone concentrations in weanling horses fed fat-supplemented or conventional diets. Concentrates were formulated to contain the same nutrient-calorie ratios and were fed with Coastal Bermuda grass hay in a 70:30 concentrate:hay ratio. Initially and at 28-d intervals the horses were weighed and measured for height, heart girth circumfer ence and subcutaneous fat thickness over the rump and ribs. Radiographs were taken of the right carpal and metacarpal phalangealjoints at the beginning and end of the experimental period for evaluation of bone density and physeal maturation. A digestion trial was conducted on each horse 10 d prior to completion of the study. On d 70, blood samples were col lected from 6 females in each treatment group over a 6-h collection period for evaluation of postprandial insulin, T₄ and T₃ concentrations in response to the diets. Horses in both groups consumed an average of 2.6% of body weight daily of total feed (Dry Matter). Weanlings consuming the fat-supplemented concentrate tended to have higher (P = .10) average daily weight gains (.80 vs .74 kg) and lower (P<.05) feed:gain ratios (6.3 vs 7.3) than weanlings fed the control diet. Gain in heart girth circumference was greater (P<.05) for the fat fed weanlings (20.1 cm) than the control weanlings (17.9 cm). All other linear measurements of growth were similar between treatments. There were no radiographic indications of abnormalities in the physes of horses on either treatment. Closure of the physes occurred at normal rates in foals on both treatments. Radiographic bone density was not different between treatments, and there was an expected increase in bone density in weanlings on both treatments over time. Digestibilities of protein and energy were similar between treatments, while ether extract and neutral detergent fiber digestibilities were higher (P<.05 and P<.06, respec tively) in those horses consuming the fat-supplemented diet (65.39 vs 57.67% and40.55 vs 35.62%, respectively). These data indicate that fat can be used as an energy source to support growth and development in weanling horses.     

Effects of season and sample handling on measurement of plasma alpha-melanocyte-stimulating hormone concentrations in horses and ponies

OBJECTIVE: To investigate effects of sample handling, storage, and collection time and season on plasma alpha-melanocyte-stimulating hormone (alpha-MSH) concentration in healthy equids. ANIMALS: 11 healthy Standardbreds and 13 healthy semiferal ponies. PROCEDURE: Plasma alpha-MSH concentration was measured by use of radioimmunoassay. Effects of delayed processing were accessed by comparing alpha-MSH concentrations in plasma immediately separated with that of plasma obtained from blood samples that were stored at 4 degrees C for 8 or 48 hours before plasma was separated. Effects of suboptimal handling were accessed by comparing alpha-MSH concentrations in plasma immediately stored at -80 degrees C with plasma that was stored at 25 degrees C for 24 hours, 4 degrees C for 48 hours or 7 days, and -20 degrees C for 30 days prior to freezing at -80 degrees C. Plasma alpha-MSH concentrations were compared among blood samples collected at 8:00 AM, 12 noon, and 4:00 PM. Plasma alpha-MSH concentrations were compared among blood samples collected in January, March, April, June, September, and November from horses and in September and May from ponies. RESULTS: Storage of blood samples at 4 degrees C for 48 hours before plasma was separated and storage of plasma samples at 4 degrees C for 7 days prior to freezing at -80 degrees C resulted in significant decreases in plasma alpha-MSH concentrations. A significantly greater plasma alpha-MSH concentration was found in September in ponies (11-fold) and horses (2-fold), compared with plasma alpha-MSH concentrations in spring. CONCLUSIONS AND CLINICAL RELEVANCE: Handling and storage conditions minimally affected plasma alpha-MSH concentrations. Seasonal variation in plasma alpha-MSH concentrations must be considered when evaluating pituitary pars intermedia dysfunction in equids.     

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.     

Alterations of growth hormone, cortisol, luteinizing hormone, and insulin concentrations in early-postnatal calves affected with diarrhea

The aim of the study was to investigate the influence of diarrheic infections during the early postnatal phase of calves on the concentrations of hormones controlling reproduction and metabolism. Blood samples were collected from 20 male and female calves via jugular vein catheters every 15 min for 6 hr at Days 3, 9, and 21 of life. The animals were classified into three groups. Group 1 (controls): healthy calves (n = 9). Group 2: calves affected with diarrhea at Day 9 (n = 7). Group 3: calves with diarrhea at Days 3 and 9 (n = 4). Infections occurred spontaneously and were mainly due to E. coli infections. All affected calves had recovered at Day 21. Mean GH concentrations in the calves in Groups 2 and 3 compared to control calves had increased by Day 3 (P<0.01; P<0.001). Cortisol levels of calves in all groups were highest at Day 3 and decreased thereafter (P<0.001). Cortisol concentrations were lower at Day 3 in animals in Groups 2 (P<0.001) and 3 (P<0.05) than in controls. Pulsatile LH release was detectable at Days 9 and 21 only in healthy calves. Insulin increased at Day 9 during diarrhea. The results indicate that cortisol concentrations decreased whereas GH concentrations were increased before diarrhea was observed. The onset of pulsatile LH release was delayed in diarrheic calves. It is concluded that diarrhea exerts effects upon the release of reproductive and metabolic hormones in early postnatal calves.     

Improvement in insulin resistance and reduction in plasma inflammatory adipokines after weight loss in obese dogs

Obesity is now a major disease of dogs, predisposing to numerous disorders including diabetes mellitus. Adipocytes are active endocrine cells, and human obesity is characterized by derangements in inflammatory adipokine production. However, it is unclear as to whether similar changes occur in dogs. The purpose of the current study was to assess insulin sensitivity and inflammatory adipokine profiles in dogs with naturally occurring obesity and to investigate the effect of subsequent weight loss. Twenty-six overweight dogs were studied, representing a range of breeds and both sexes. All dogs underwent a weight loss program involving diet and exercise. Body fat mass was measured by dual-energy x-ray absorptiometry; plasma concentrations of insulin, glucose, and a panel of inflammatory adipokines (including acute-phase proteins, cytokines, and chemokines) were also analyzed. Body fat mass before weight loss was positively correlated with both plasma insulin concentrations (Kendall τ = 0.30, P = 0.044) and insulin:glucose ratio (Kendall τ = 0.36, P = 0.022), and both decreased after weight loss (P = 0.0037 and 0.0063, respectively). Weight loss also led to notable decreases in plasma tumor necrosis factor-α (TNF-α), haptoglobin, and C-reactive protein concentrations (P < 0.05 for all), suggesting improvement of a subclinical inflammatory state associated with obesity. This study has demonstrated that in obese dogs, insulin resistance correlates with degree of adiposity, and weight loss improves insulin sensitivity. Concurrent decreases in TNF-α and adipose tissue mass suggest that in dogs, as in humans, this adipokine may be implicated in the insulin resistance of obesity.