Changes in plasma ghrelin and growth hormone concentrations in mature Holstein cows and three-month-old calves

We measured changes in plasma ghrelin and GH concentrations in mature Holstein cows and 3-mo-old female Holstein calves fed at scheduled times. Our objective was to determine the characteristics of ghrelin secretion in dairy cattle and its influence on GH. Animals were fed at 0800 and 1600 for 2 wk before and during experiments. Plasma was sampled for 24 h at 2-h intervals in Exp. 1. In mature cows, plasma ghrelin concentrations decreased (P < 0.01) just after 0800 but not at the 1600 feeding. Ghrelin concentrations were lower (P < 0.01) in calves than in mature cows and they did not decrease after feeding in calves. The temporal relationship between ghrelin and GH remained unclear. In Exp. 2, plasma was sampled 2 h before and after both morning and evening feedings at 20-min intervals. Plasma ghrelin concentrations decreased (P < 0.05) 40 min after 0800 feeding and 60 min after 1600 feeding in mature cows. These results indicate that in mature cows, plasma ghrelin concentration decreased after feeding, but this decrease was not evident in 3-mo-old calves. Further studies are required to define the relationship between plasma ghrelin and GH concentrations.     

Active immunization against ghrelin decreases weight gain and alters plasma concentrations of growth hormone in growing pigs

Ghrelin has been implicated in the control of food intake and in the long-term regulation of body weight. We theorize that preventing the ability of ghrelin to interact with its receptors, would eventually lead to decreased appetite and thereby decrease body weight gain. To test our hypothesis, pigs were actively immunized against ghrelin. Ghrelin((1-10)) was conjugated to BSA and emulsified in Freund's incomplete adjuvant and diethylaminoethyl-dextran. Primary immunization was given at 19 weeks of age (WOA), with booster immunizations given 20 and 40 days after primary immunization. Body weight (BW) and plasma samples were collected weekly beginning at 19 WOA, and feed intake was measured daily. Fourteen days after primary immunization, the percentage of bound (125)I-ghrelin in plasma from immunized pigs was increased compared with control animals (P<0.001). Voluntary feed intake was decreased more than 15% in animals that were actively immunized against ghrelin compared with controls. By the end of the experiment, immunized pigs weighed 10% less than control animals (P<0.1). Concentrations of GH were increased (P<0.05) in immunized pigs. Apoptosis was not observed in post-mortem samples obtained from the fundic region of the stomach. Our observations suggest that immunization against ghrelin induces mild anorexia. This procedure could potentially be used as a treatment to control caloric intake and obesity.     

Effects of intravenous ghrelin injection on plasma growth hormone,insulin and glucose concentrations in calves at weaning

Ghrelin action, which stimulates growth hormone (GH) secretion, may alter during the weaning period in calves. Our objective was to compare the effects of intravenous ghrelin injection on plasma GH, insulin and glucose concentrations in calves around the weaning period. Four Holstein bull calves were fed whole milk and allowed free access to solid feeds, and weaned at 7 weeks of age. Measurements were performed at weeks 1, 2, 4, 6, 7, 9, 11 and 13, when calves were intravenously injected with ghrelin (1.0 μg/kg body weight (BW)) through a catheter, and jugular blood samples were obtained temporally relative to the injection time. Estimated digestible energy intake per metabolic BW transiently decreased at week 7 because of low solid intake immediately after weaning, and thereafter gradually increased. Plasma insulin and glucose concentrations were not affected by ghrelin injection at all ages. In contrast, plasma GH concentrations increased with ghrelin injection at all ages. The incremental area of GH at week 7 was greatest and significantly higher compared with weeks 2, 4, 6 and 9. This result suggests that nutrient insufficiency immediately after weaning enhances GH responsiveness to ghrelin.     

Salivary secretory IgA concentrations in beagle dogs

The normal concentrations of salivary secretory IgA (sIgA) were examined, and the response of sIgA to acute stress was evaluated in dogs. Ten clinically healthy beagle dogs familiarized with the method of saliva sampling were used. During the non-stress period, saliva samples were collected between 0800 hr and 1700 hr at 1-hr intervals for 7 consecutive days and analyzed for sIgA concentration. After a 1-day control period, a noise stressor was presented for 15 min between 0845 hr and 0900 hr on 2 consecutive days. Saliva was collected at pre-stress, immediately after, 30 min after and 60 min after the stress. The average sIgA concentration over the 2-day period was compared with the control value. Environmental stimuli were restricted. During the non-stress period, significant variations were observed during the diurnal pattern, in which sIgA increased in the morning and then decreased; and the day-to-day variations were significant except at 0800 hr and 0900 hr. During the stress experiments, the sIgA concentration decreased significantly, immediately after and 30 min after the noise stress, and then increased to the same level as the control value by 60 min after the stress. When estimating the effectiveness of salivary sIgA as a marker of stress in dogs, the appropriate time for saliva sampling appears to be in the morning. Salivary sIgA was deemed potentially useful as a marker of stress in dogs.     

Effects of food intake and food withholding on plasma ghrelin concentrations in healthy dogs

OBJECTIVE: To investigate the physiologic endocrine effects of food intake and food withholding via measurement of the circulating concentrations of acylated ghrelin, growth hormone (GH), insulin-like growth factor-I (IGF-I), glucose, and insulin when food was administered at the usual time, after 1 day's withholding, after 3 days' withholding and after refeeding the next day in healthy Beagles. ANIMALS: 9 healthy Beagles. PROCEDURES: Blood samples were collected from 8:30 AM to 5 PM from Beagles when food was administered as usual at 10 AM, after 1 day's withholding, after 3 days' withholding, and after refeeding at 10 AM the next day. RESULTS: Overall mean plasma ghrelin concentrations were significantly lower when food was administered than after food withholding. Overall mean plasma GH and IGF-I concentrations did not differ significantly among the 4 periods. Circulating overall mean glucose and insulin concentrations were significantly higher after refeeding, compared with the 3 other periods. CONCLUSIONS AND CLINICAL RELEVANCE: In dogs, food withholding and food intake were associated with higher and lower circulating ghrelin concentrations, respectively, suggesting that, in dogs, ghrelin participates in the control of feeding behavior and energy homeostasis. Changes in plasma ghrelin concentrations were not associated with similar changes in plasma GH concentrations, whereas insulin and glucose concentrations appeared to change reciprocally with the ghrelin concentrations.     

Effects of chronic obesity and weight loss on plasma ghrelin and leptin concentrations in dogs

The objective of this study was to evaluate, in dogs, the effects of obesity and weight loss on plasma total ghrelin and leptin concentrations. Twenty-four Beagle dogs, 12 control lean and 12 obese dogs of both genders and aged between 1 and 9 years, were used for the experiments. Mean body weight was 12.7+/-0.7 kg for the lean group and 21.9+/-0.8 kg for the obese group. The trial was divided into three phases. During phase 1, all 24 Beagle dogs were fed a maintenance diet. During phase 2, the obese dogs were submitted to a weight loss protocol with a high protein-low energy diet. The weight loss protocol ended once dogs reached optimal body weight. During phase 3, the dogs that were submitted to the weight loss protocol were maintained at their optimal body weight for 6 months. Plasma total ghrelin, leptin, insulin and glucose concentrations were measured to evaluate the effects of obesity and weight loss on these parameters in dogs. Body weight, body condition score, thoracic and pelvic perimeters, and ingested food amounts were also recorded during the study. Obese dogs demonstrated a significant decrease in plasma ghrelin and a significant increase in plasma leptin and insulin concentrations when compared with control dogs. During weight loss, significant increases in plasma total ghrelin and glucose and significant decreases in plasma leptin and insulin were observed. The increase in plasma ghrelin concentrations seemed to be transient. Body weight and the morphometric parameters correlated positively with leptin concentrations and negatively with total ghrelin concentrations. These results suggest that ghrelin and leptin could play a role in dogs in the adaptation to a positive or negative energy balance, as observed in humans.     

Effects of fasting and refeeding on plasma concentrations of leptin, ghrelin, insulin, growth hormone and metabolites in swine

The effects of nutritional status, such as fasting and refeeding, on leptin and ghrelin secretion in swine were examined. The swine (n = 4) were fasted for 54 h and plasma hormone levels were measured before, during and after fasting. Plasma leptin and insulin concentrations began to decrease 12 and 6 h into the fasting period, respectively (P < 0.05), and maintained a low level for the remaining period of fasting. Plasma leptin and insulin returned to the pre‐fasting value 6 and 12 h after refeeding, respectively. Plasma ghrelin concentrations showed a nocturnal periodicity during the fasting period; it increased nocturnally at 36 and 42 h into the fasting period (P < 0.05). Plasma growth hormone levels did not show any remarkable changes during the fasting. Plasma glucose levels showed a modest fall during fasting and significantly decreased (P < 0.05) at 24 h into the fasting period, returning to pre‐fasting levels after refeeding. Plasma nonesterified fatty acid levels increased (P < 0.05) at 12 h into the fasting period and returned to the pre‐fasting level 6 h after refeeding. These results indicate that plasma leptin, insulin and ghrelin play an important role in maintaining energy homeostasis in swine. The plasma ghrelin did not continuously increase, but showed nocturnal periodicity during fasting. This may suggest that ghrelin is also involved in physiological processes other than energy homeostasis.     

Negative energy balance increases periprandial ghrelin and growth hormone concentrations in lactating dairy cows

The reported effects of feeding on growth hormone (GH) secretion in ruminants have been inconsistent, and are likely influenced by energy status of animals. High-producing dairy cows in early lactation and late lactation were used to assess the effects of energy balance on temporal variation of plasma metabolites and hormones. Cows were fed a single diet once daily, and feed was withdrawn for 90 min prior to feeding. Beginning at the time of feed withdrawal, plasma samples were collected via jugular catheters hourly for 24h. Concentrations of non-esterified fatty acids and GH were measured for all samples, while insulin, glucose, and acylated (active) ghrelin were quantified for four sample times around feeding. As expected, calculated energy balance was significantly lower in early lactation than late lactation cows (-43.5 MJ retained/day versus 7.2 MJ retained/day). Following the primary meal of the day, a GH surge was observed in early lactation but not in late lactation cows. This difference was not explained by temporal patterns in non-esterified fatty acid, insulin, or glucose concentrations. However, a preprandial ghrelin surge was observed in early lactation only, suggesting that ghrelin was responsible for the prandial GH surge in this group. Results of a stepwise regression statistical analysis showed that both preprandial ghrelin concentration and energy balance were significant predictors of prandial GH increase over baseline. Adaptations to negative energy balance in lactating dairy cattle likely include enhanced ghrelin secretion and greater GH response to ghrelin.     

Effects of racing and nontraining on plasma thyroid hormone concentrations in sled dogs

OBJECTIVE: To determine the effects of racing and nontraining on plasma thyroxine (T4), free thyroxine (fT4), thyroid-stimulating hormone (TSH), and thyroglobulin autoantibody (TgAA) concentrations in sled dogs and compare results with reference ranges established for dogs of other breeds. DESIGN: Cross-sectional study. ANIMALS: 122 sled dogs. PROCEDURE: Plasma thyroid hormone concentrations were measured before dogs began and after they finished or were removed from the Iditarod Trail Sled Dog Race in Alaska and approximately 3 months after the race. RESULTS: Concentrations of T4 and fT4 before the race were less than the reference range for nonsled dogs in 26% and 18% of sled dogs, respectively. Immediately after racing, 92% of sled dogs had plasma T4 concentrations less than the reference range. Three months after the race, 25% of sled dogs had plasma T4 concentrations less than the reference range. For T4, fT4, TSH, and TgAA, significant differences were not detected in samples collected before the race versus 3 months later. CONCLUSIONS AND CLINICAL RELEVANCE: Plasma T4, fT4, and TSH concentrations decreased in dogs that complete a long distance sled dog race. Many clinically normal sled dogs have plasma T4 and fT4 values that are lower than the reference range for nonsled dogs. We suggest that the reference ranges for sled dogs are 5.3 to 40.3 nmol/L and 3.0 to 24.0 pmol/L for plasmaT4 and fT4 concentrations, respectively, and 8.0 to 370 mU/L for TSH.     

Variations in plasma growth hormone concentrations in laying hens.

1. Variations in plasma concentrations of growth hormone (GH) and luteinising hormone (LH) were measured by radioimmunoassay in blood samples taken during the ovulatory cycle of the domestic hen. 2. A peak in LH concentrationas observed 4 h prior to ovulation. 3. Plasma GH concentrations were quite variable but a significant increase was observed between 2 h before and 2 h after ovulation and the concentration remained elevated for about 4 h thereafter.     

Relationship between testicular transferrin and plasma estradiol-17beta concentrations of dogs with azoospermia and dogs with sertoli cell tumors.

Testicular Transferrin (Tf) and peripheral plasma estradiol-17beta (E2) concentrations were measured in 3 dogs with azoospermia (AZ dogs), 3 dogs with Sertoli cell tumors (SC dogs), and 5 normal male Beagles. The mean Tf concentrations in the testes of the AZ dogs and the affected testes of the SC dogs, and the plasma E2 concentrations in both these groups of dogs were significantly higher than the values in normal dogs (P<0.05, 0.01 and 0.01, respectively). Therefore, excessive E2 secretion by hyperfunctioning Sertoli cells is thought to have caused the azoospermia in the 3 dogs.     

Effect of octreotide on plasma concentrations of glucose, insulin, glucagon, growth hormone, and cortisol in healthy dogs and dogs with insulinoma

The inhibitory effect of the somatostatin analogue octreotide on the secretion of insulin could be used in the treatment of insulinoma. However, current information on the effectiveness of octreotide in dogs is conflicting. Therefore, the endocrine effects of a single subcutaneous dose of 50 microg octreotide were studied in healthy dogs in the fasting state (n=7) and in dogs with insulinoma (n=12). Octreotide did not cause any adverse effects. In healthy dogs in the fasting state, both plasma insulin and glucagon concentrations declined significantly. Basal (non-pulse related) GH and ACTH concentrations were not affected. A slight but significant decrease in the plasma glucose concentrations occurred. Dogs with insulinoma had significantly higher baseline insulin concentrations and lower baseline glucose concentrations than healthy dogs in the fasting state. Plasma glucagon, GH, ACTH, and cortisol concentrations did not differ from those in healthy dogs. Baseline plasma insulin concentrations decreased significantly in dogs with insulinoma after octreotide administration, whereas plasma concentrations of glucagon, GH, ACTH, and cortisol did not change. In contrast to the effects in the healthy dogs, in the dogs with insulinoma plasma glucose concentrations increased. Thus, the consistent suppression of plasma insulin concentrations in dogs with insulinoma, in the absence of an suppressive effect on counter-regulatory hormones, suggests that further studies on the effectiveness of slow-release preparations in the long-term medical treatment of dogs with insulinoma are warranted.     

Relationship between plasma insulin and triglyceride concentrations in hypertriglyceridaemic donkeys

Hyperinsulinaemia is a commonly-observed characteristic of insulin resistance, and a reduction in insulin sensitivity is thought to be either a causative and/or symptomatic feature of equine hyperlipaemia. A positive correlation (r=0·545, P=0·0015) existed between plasma insulin and triglyceride concentrations determined in 31 donkeys with naturally occurring hyperlipidaemia/hyperlipaemia. Greater insulin values tended to occur in the animals with an overweight body score. Inter-animal variation in insulin concentrations, however, prevented the identification of any differences either within hypertriglyceridaemic donkeys (when classified by clinical condition, date of arrival to a sanctuary and eventual outcome after treatment) or between groups of normotriglyceridaemic (n=6) and experimentally fasted hypertriglyceridaemic (n=5) donkeys. Determination of basal plasma insulin concentrations may not provide an accurate assessment of underlying insulin sensitivity. Alternatively, hyperinsulinaemia may be evident only in animals with established insulin resistance.     

Relationship between the serum concentrations of serotonin and lipids and aggression in dogs

The serum concentrations of serotonin and lipids--triglycerides, total cholesterol, low density lipoprotein, high density lipoprotein and very low density lipoprotein--were determined in 18 normal dogs and 23 dogs diagnosed as aggressive on the basis of interviews with their owners and an assessment of their behaviour with reference to a canine overt aggression chart. The serum serotonin levels in the aggressive dogs were significantly lower than in the normal dogs (P<0.01), but the differences in serum lipids between the two groups were not statistically significant.     

Relationship between plasma creatinine concentration and glomerular filtration rate in dogs

Glomerula filtration rate (GFR), plasma creatinine concentration (CR), and plasma urea nitrogen concentration (BUN) were measured in 129 adult dogs with reduced renal mass. A preliminary examination of the relationship between CR and GFR was conducted, and the inverse model (GFR vs. 1/CR) was chosen for further evaluation. The slope of the regression of GFR on 1/CR which was computed from actual data was not statistically different from a theoretical regression line generated from the clearance equation.
Evaluation of subsets of the population revealed no significant difference between male ( n = 69) and female (n = 60) dogs on the slope of the regression equations. Diets differing in protein concentration (16% protein, n = 35: 21% protein, n = 75: 32% protein, n = 19) did not cause a significant difference in the slope of the regression equations.
The regression equation and the confidence intervals generated in this study may be used to predict a probable range of GFR values from CR in individual dogs. Such values may be useful in adjusting drug dosages in dogs with renal disease. However, since the derived equation did not differ significantly from the theoretical inverse relationship between GFR and CR, it remains to be established whether the equation is advantageous.     

Relationship of disease progression and plasma histamine concentrations in 11 dogs with mast cell tumors

Plasma histamine concentrations (PHCs) were measured serially over 9 months or until death in 11 dogs with mast cell tumors (MCTs). Eight dogs had grossly visible disease and the other 3 dogs had microscopic disease. Initial PHCs in the dogs with gross disease were significantly higher than PHCs in healthy dogs (median, 0.73 ng/mL and 0.19 ng/mL respectively; P < .009), whereas initial PHCs in dogs with microscopic disease showed no difference from controls. Seven dogs subsequently had progressive increases in PHC, and developed hyperhistaminemia (median, 14.0 ng/mL; range, 5.11-30.1 ng/nL). These 7 dogs died from MCTs, and 1 had general weakness with rapid lysis of a large tumor burden after radiation therapy. PHCs of the other 4 dogs were less than 1 ng/mL during the study. These 4 dogs were still alive with adequate control of the tumor at the conclusion of the study. Four of the 11 dogs initially had gastrointestinal (G1) signs, which abated soon after administration of histamine-2 (H-2) blockers. No significant difference was found between PHCs in dogs with GI signs and those without GI signs (median, 0.86 ng/mL and 0.35 ng/mL. respectively). Thereafter, 7 dogs had serious GI complications for which H-2 blocker therapy was ineffective. PHCs in these 7 dogs were extremely high (median, 12.2 ng/mL; range, 3.42-30.1 ng/nL). Results of this study demonsrated that PHC was one factor related to disease progression, and indicated that marked hyperhistaminemia was associated with the GI signs refractory to H-2 blocker therapy in dogs with MCTs.     

Developmental changes in the regulation of plasma growth hormone concentrations in Holstein calves

A study was initiated to determine whether development of a functional ruminant digestive system was associated with alterations in plasma growth hormone (GH) concentration. Holstein bull calves were fed milk or milk with grain until studied at the age of 1 month (n = 12). Calves placed on pasture with some grain supplementation were studied at the age of 3 months (n = 6) to determine plasma GH concentration in an animal with fully developed ruminant metabolism. Blood samples were taken at 10-minute intervals for 5 hours, followed by administration of bovine GH-releasing factor (0.075 micrograms/kg of body weight) and subsequent blood sample collection for 1 hour. On the following day, a blood sample was collected via jugular cannula, clonidine (10 micrograms/kg) was administered, and blood samples were subsequently obtained. Data indicated that milk-fed calves had higher mean plasma GH concentration than did either milk/grain-fed or older calves. The difference in mean plasma GH concentration was related to higher secretory pulse amplitude. Pituitary responses to bovine GH-releasing factor did not differ among the 3 groups, but response to clonidine were greater in milk-fed calves than in calves of the other groups. These data indicate that the change from a nonruminant to a ruminant-type gastrointestinal tract, perhaps attributable to subsequent changes in metabolism, may induce changes in hypothalmic function to decrease GH concentration.