Interrelationships of zinc with glucose and insulin metabolism in humans

Hyperzincemia has been reported to cause alterations in the homeostasis of glycid metabolism. To determine this effect on plasma glucose and insulin levels, we studied 36 normal individuals of both sexes aged 22–26 y after a 12-h fast. The tests were initiated at 7:00am when an antecubital vein was punctured and a device for infusion was installed and maintained with physiological saline. Zinc was administered orally at 8:00am. Subjects were divided into an experimental group of 22 individuals who received doses of 25, 37.5, and 50 mg of zinc and a control group of 14 individuals. Blood samples were collected over a period of 240 min after the basal samples (−30 and 0 min). We did not detect any change in plasma glucose or insulin levels, a fact that we attribute either to the ineffectiveness of the 50 mg dose of zinc or to the lack of human response to the acute action of this trace element. The individuals who ingested zinc showed a significant fall in plasma cortisol, probably caused by the action of this trace element.     

Lack of acute zinc effects in glucose metabolism in healthy and insulin-dependent diabetes mellitus patients

Acute or chronic zinc administration may cause hyperglycemia in experimental animals. These findings are attributed to permissive actions of glucocorticoids and glucagon upon hepatic gluconeogenesis and glycogenolysis. The effect of Zn⁺⁺ on plasma glucose, C-peptide, glucagon, and cortisol was investigated in healthy and insulin-dependent diabetes mellitus (IDDM) patients. Ten normal individuals (5 of each sex, aged 24.10 ± 1.96) and 10 IDDM (5 of each sex, aged 25.20 ± 8.10) were tested at 7:00 AM after 12-h fast. Twenty-five mg of Zn⁺⁺ were administered intravenously during 1 min, and blood samples were collected from the contralateral arm at 0, 3, 30, 60, 90 and 120 min after Zn⁺⁺ injection. The plasma levels of glucose, C-peptide, and glucagon remained constant throughout the experimental period in both groups studied. Plasma cortisol levels decreased significantly, which is consistent with our previous findings. These results suggest that, in contrast to experimental animals, acute Zn⁺⁺ administration, despite decreasing cortisol levels, does not change carbohydrate metabolism in human beings.     

Interaction among zinc,glucose, and insulin in normal individuals during glucose and tolbutamid perfusion

Reports in the literature have shown that acute or chronic zinc administration may cause hyperglycemia, with a fall in serum or insular insulin occurring in experimental animals. On the other hand, under conditions of both acute and chronic hyperglycemia, an increase, a decrease, or a normal level of blood zinc has been observed in studies conducted on humans. Thus, the objective of the investigation described here was to determine the relationship existing among zinc, glucose, and insulin under acute conditions. Thirty-six subjects of both sexes (mean age, 23 yr) were tested at 7:00A.M. after a 12-h fast. Two antecubital veins of both forearms were punctured and maintained with physiological saline. Three experiments were performed in which zinc was administered orally, and hypertonic glucose and tolbutamid were administered intravenously. Blood samples were then collected over a period ranging from 93 to 240 min after the basal times of −30 and 0 min. Hyperzincemia did not cause changes in plasma glucose or insulin either in the absence of or during perfusion of glucose. Hyperglycemia, hypoglycemia, and hyperinsulinemia did not modify serum zinc levels. These results demonstrate that acute zinc administration did not change carbohydrate metabolism and that sudden variations in glucose and insulin levels did not modify the serum profile of zinc.     

Influence of long-term exposure to an air-conditioned environment on the diurnal cortisol rhythm

In the present study, the influence of the long-term use of air-conditioning in summer on the cortisol rhythm was examined by measuring the rhythm in subjects who had been exposed to air-conditioning for a short [S] or long [L] time. Investigations were conducted twice in July and September. Atmospheric temperature and relative humidity near the subjects were measured for three days in each season. Saliva samples for cortisol analysis were collected every 2 hours during the daytime beginning at 8:00 h with subsequent sampling times at 10:00, 12:00, 14:00, 16:00, 18:00, 20:00 and 22:00 h. A questionnaire on sleep and duration of air-conditioning use was also undertaken. Ambient mean temperature was higher in the S group (mean+/-SD; 30.8+/-1.2 degrees C in July, 28.0+/-0.8 degrees C in September) than in the L group (28.0+/-1.2 degrees C in July, 27.3+/-1.0 degrees C in September) (p<0.01), while mean relative humidity did not differ. There were no differences in bedtime, waking time and sleeping hours either between groups or months. Diurnal patterns of salivary cortisol rhythm in July and September were similar in the S group, but the L group had a delayed rise of morning cortisol secretion in September compared with July. These results suggest that long-term exposure to an air-conditioned environment might adversely affect the human cortisol rhythm.     

Atrial natriuretic factor inhibits the CRH-stimulated secretion of ACTH and cortisol in man.

Corticotrophic secretion of ACTH is stimulated by corticotropin-releasing hormone (CRH) and arginine vasopressin (AVP), and suppressed by glucocorticoids. In vitro and preclinical studies suggest that atrial natriuretic factor (ANF) may be a peptidergic inhibitor of pituitary-adrenocortical activity. The aim of this study was to elucidate a possible role of ANF as a modulator of ACTH release in humans. A bolus injection of 100 micrograms human CRH (hCRH) during a 30 min intravenous infusion of 5 micrograms/min human alpha atrial natriuretic factor (h alpha ANF) was administered at 19:00 to six healthy male volunteers. In comparison to saline, a blunted CRH-stimulated secretion of ACTH (mean maximum plasma level +/- SD 45 min after hCRH: saline 46.2 +/- 14.2 pg/ml, h alpha ANF 34.6 +/- 13.8 pg/ml, p-value = 0.007) and a delayed rise (10 min) in cortisol were detected. The maximum plasma cortisol levels remained nearly unchanged between saline and h alpha ANF administration (mean maximum plasma level +/- SD 60 min after hCRH: saline 182 +/- 26 ng/ml, h alpha ANF 166 +/- 54 ng/ml). No effects of h alpha ANF on basal cortisol levels were observed; in contrast, basal ACTH plasma levels were slightly reduced. Basal blood pressure and heart rate remained unaffected. In the control experiment, infusion of 3 IU AVP in the same experimental paradigm increased basal and stimulated ACTH and cortisol levels significantly in comparison to saline. These observations suggest that intravenously administered haANF inhibits the CRH-stimulated release of ACTH in man.     

Role of prostaglandin E2 (PGE2) on the corticotropin-releasing hormone (CRH)-induced ACTH release in healthy men.

The effect of PGE2 on ACTH and cortisol responses to CRH was studied in 6 healthy men who received CRH i.v. during either saline or PGE2 infusions which were started 60 min. before testing. ACTH and cortisol responses to CRH were greater during PGE2 infusion compared to the control study. The results indicate that PGE2 positively modulates CRH-induced ACTH secretion.     

Cortisol and GH secretory dynamics, and their interrelationships, in healthy aged women and men

We studied 130 healthy aged women (n = 57) and men (n = 73), age 65-88 yr, with age-related reductions in insulin-like growth factor I and gonadal steroid levels to assess the interrelationships between cortisol and growth hormone (GH) secretion and whether these relationships differ by sex. Blood was sampled every 20 min from 8:00 PM to 8:00 AM; cortisol was measured by RIA and GH by immunoradiometric assay, followed by deconvolution analyses of hormone secretory parameters and assessment of approximate entropy (ApEn) and cross-ApEn. Cortisol mass/burst, cortisol production rate, and mean and integrated serum cortisol concentrations (P < 0.0005), and overnight basal GH secretion (P < 0.05), were elevated in women vs. men. Integrated cortisol concentrations were directly related to most measures of GH secretion in women (P < 0.01) and with mean and integrated GH concentrations in men (P < 0.05). Integrated GH concentrations were directly related to mean and integrated cortisol levels in women (P < 0.005) and men (P < 0.05), with no sex differences. There were no sex differences in cortisol or GH ApEn values; however, the cross-ApEn score was greater in women (P < 0.05), indicating reduced GH-cortisol pattern synchrony in aged women vs. men. There were no significant relationships of integrated cortisol secretion with GH ApEn, or vice versa, in either sex. Thus postmenopausal women appear to maintain elevated cortisol production in patterns that are relatively uncoupled from those of GH, whereas mean hormone outputs remain correlated.     

Zinc: an inhibitor of prolactin (PRL) secretion in humans

The response of plasma prolactin (PRL) to oral administration of increasing doses of zinc (25.0, 37.5 and 50.0 mg) was studied in 17 normal adult men and women. Blood samples were collected at 10 and 30-min intervals over a period of 120 min after two basal times (-30 and 0 min). PRL concentrations significantly fell below basal levels in all subjects in response to the increase in plasma zinc levels, as compared to the controls. These results suggest that acute hyperzincemia can inhibit basal PRL secretion in normal individuals.     

Detailed examination of episodic bursts of rGH secretion by high frequency blood sampling in normal male rats

Episodic bursts of rat growth hormone (rGH) secretion were examined in detail by monitoring circulating rGH concentration every 2 min (70 ul blood samples were taken). Conscious free moving normal male rats implanted with indwelling cannulae were used. Between 11:00–13:00 h and between 14:00–15:20 h, basal levels of rGH were found to be 30–50 ng/ml. Bursts of rGH secretion had many different shapes, including a “single spike”, “compound peak” and “plateau” but no regularity or periodicity of changes of rGH concentration was observed. Therefore we could not apply basic statistical treatments such as the standard deviation or standard error of means which are based on the assumption of normal distributions. When the downward slopes of each major spikes comprised more than 3 sequential samples, the decay slope was analyzed. Of 16 slopes, 10 were parallel with a theoretical curve of a 7 min half-life (rGH half-life is 5–7 min) indicating that no more rGH was secreted during the period of the downward slope. The slopes with significantly slower disappearnace rates can be interpreted as indicating that a significant amount of rGH is secreted during the period of the downward slopes. Our experimental design revealed a more complex pattern of rGH secretory bursts than had hitherto been described. Our technique revealed more physiological characteristics of rGH secretion by more frequent sampling of very small volumes of blood. This technique eliminated unnecessarily excessive bleeding and avoided the necessity of returning red blood cells.     

Acute effects of hydrocortisone on circulating growth hormone levels in patients with acromegaly.

Aim of our study was to investigate the acute effects of intravenous infusion of hydrocortisone on circulating growth hormone (GH) levels in acromegaly. We studied 5 adult patients with active acromegaly, 3 males and 2 females; age 52 +/- 3.6 years, body mass index 27 +/- 1 kg/m2. The patients underwent in randomized order from 0 to 120 min: (1) intravenous infusion of saline, 250 ml; (2) bolus intravenous injection of hydrocortisone succinate, 100 mg at time 0 followed by intravenous infusion of hydrocortisone succinate, 250 mg in 250 ml of saline for 120 min. Blood samples for GH, cortisol and glucose assay were taken at -15, 0 (time of beginning of saline or hydrocortisone infusion), 15, 30, 45, 60, 90, 120, 150 and 180 min. In all the acromegalic patients, during hydrocortisone succinate infusion, GH values clearly fell with respect to saline (nadir range 18.4-50.5% with respect to baseline levels) with nadir between 60 and 180 min after the beginning of the infusion. Our data show that acute and sustained hypercortisolism, decreases circulating GH levels in acromegaly. It seems likely that also in acromegalic patients as well as in normal subjects short-term increases in serum cortisol levels may be able to cause an enhancement of hypothalamic somatostatin secretion, which in turn may be responsible for the glucocorticoid-mediated GH inhibition.     

Feedback suppression of ACTH secretion by cortisol in dogs: lags after large signals equal those following very small signals

Previously, low stepwise infusions of cortisol in resting adrenalectomized dogs (plateaux less than or equal to 6 micrograms/dL) were shown to reduce ACTH secretion only after 20 min. In the present study, large, steep-sloped cortisol signals were used to try to evoke faster feedback. Adrenalectomized male mongrel dogs were maintained on exogenous steroids until 48 h before the experiment. Of the 23 experiments on 15 dogs (under light pentobarbital anesthesia), 12 were on resting dogs, 7 on dogs stressed by variable insulin infusion (keeping plasma glucose at 18-40 mg/dL), and 4 stressed as above but with 4 h of low cortisol infusion (plasma congruent to 5 micrograms/dL) before applying the feedback signal. After a 50-min control period, a 30-min feedback period was initiated by one of two i.v. cortisol signals: (a) injection of 0.3 mg/kg or (b) infusion of 46 micrograms kg-1 min-1. Both raised plasma cortisol above physiological limits (within 2 and 6 min, respectively). In each experiment, 23 timed venous blood samples were assayed for plasma ACTH and cortisol. ACTH secretion rates were calculated continuously using a validated single-compartment method. Results from both types of cortisol signals were indistinguishable, and were thus pooled. In the unstressed dogs, control-period ACTH secretion of 0.97 +/- 0.12 mU kg-1 min-1 showed no significant decline due to the feedback signal for 20.3 +/- 1.4 min.(ABSTRACT TRUNCATED AT 250 WORDS)     

Influence of acoustic stimulation on the circadian and ultradian rhythm of premature infants

The aim of the present study was to evaluate the development of the circadian rhythm of the salivary cortisol in premature infants and its correlation with the onset of the sleep–activity behavior pattern during the first 3 weeks of life under controlled light:dark conditions. Furthermore, we investigated the influence of acoustic stimulation by audiotaped lullabies or the maternal voice on the cortisol values and long-term sleep–activity patterns. The study was a block-randomized, prospective clinical trial with a study population of 62 preterm neonates (30?<?37 gestational age). We compared two study groups who listened either to music or to the maternal voice (music: N?=?20; maternal voice: N?=?20) with a matched control group (N?=?22). The acoustic stimulation took place every evening between 20:00 and 21:00?h for 30?min over a period of 2 weeks. The cortisol values and activity–rest behavior of the neonates were determined during the first 3 weeks of life on the 1st, 7th and 14th day. Actigraphic monitoring was used to record the activity pattern continuously over 24?h and a validated algorithm for neonates was used to estimate sleep and wakefulness. The saliva samples were obtained 10?min before and 10?min after the acoustic interventions for the study groups. Additionally, saliva samples were obtained from the control group seven times over a 24-h period (20:00, 21:00, 01:00, 05:00, 08:00, 13:00 and 17:00?h). The cortisol data were analyzed by fast Fourier transformation to assess periodic characteristics and frequencies. Hierarchical linear modeling was further performed for the statistical analysis. Results: The cortisol rhythm analysis indicated a circadian rhythm pattern for only one premature infant, all others of the neonates showed no circadian or ultradian rhythm in cortisol. Cortisol level of the premature neonates was significantly higher during the first day of the study period at night-time (median: 17.1?nmol/L, IQR?=?9.7–24.4?nmol/L) than on days 7 (median: 9.6?nmol/L, IQR?=?4.7–14.6?nmol/L; Tukey-HSD, z?=?4.12, p?<?0.001) and 14 (IQR?=?5.8–13.7?nmol/L; Tukey-HSD, z?=?2.89, p?<?0.05). No significant effect of acoustic stimulation was observed on the cortisol concentration and sleep–wake behavior. The activity–sleep rhythm of preterm neonates was dominated by ultradian rhythm patterns with a prominent period length of 4?h (30.5%). Activity frequencies of neonates were also significantly higher overnight on the first study day (mean: 329?±?185.1?U) than of night seven (mean: 260.2?±?132.4?U; Tukey-HSD, z?=?2.50, p?<?0.05). Quiet-activity patterns increased, whereas high-activity patterns decreased during the observation period. Average sleep time increased significantly during the study time from day 1 to day 7 (Tukey-HSD, z?=?2.51, p?<?0.05). In conclusion, premature infants showed higher cortisol levels – without a circadian rhythmicity – and higher activity frequencies in the first days after birth which may reflect an adaptation process of neonates after birth. Cortisol concentrations and the activity patterns were not influenced by music interventions.     

Diurnal variations in plasma ACTH, cortisol and beta-endorphin levels in cocaine addicts.

In order to establish possible alterations in the hypothalamic pituitary-adrenal axis and in ACTH-related opioids in cocaine addicts, plasma ACTH, cortisol and beta-endorphin levels were measured throughout the day in 9 cocaine addicts [age: 27 +/- 5 years (mean +/- SE); weight: 72 +/- 6.1 kg, duration of cocaine addiction: at least 2 years] on the day of their admission to a recovery community for drug abusers (first test) and after 15 days of abstinence (second test). Nine normal controls (age: 28 +/- 6 years; weight: 73 +/- 3.2 kg) were tested only once in a similar manner. Blood samples were taken at 06:00, 08:00, 10:00, 12:00, 18:00 and 20:00 h and served for hormonal assays. Urine samples were taken from cocaine addicts at 08:00 h on the experimental day and on the following day. Results of both urine assays were positive for cocaine catabolites, indicating cocaine administration during the day before the experimental test. From the day of their admission in the community (1st experimental day), the patients were forbidden to use cocaine. For 4 days after admission, they were treated with symptomatics to attenuate withdrawal symptoms. Thereafter, the patients underwent a washout period of pharmacological treatments for 10 days before being retested (second test). Urine samples taken at 08:00 h on this second experimental day and on the next day were negative for the presence of drug catabolites. During the first test, cocaine addicts showed higher plasma ACTH, cortisol and beta-endorphin levels than normal controls at all examined time points.(ABSTRACT TRUNCATED AT 250 WORDS)     

Plasma cortisol in cattle: Circadian rhythm and exposure to a simulated high altitude of 5,000 m

After a 2 week control period at 400 m, cattle were exposed to 5,000 m simulated altitude for 2 weeks, which was followed by a 2-week post-altitude control period. Plasma cortisol values from blood samples taken every 30 min for a total of 24 h indicated that cortisol was secreted episodically and that a circadian rhythm existed. When cortisol values were grouped into 4, 6-h periods, plasma cortisol was most abundant from 06:00 to 12:00 h with an average of 0.96µ g/100 ml and least abundant from 00:30 to 06:00 h with an average of 0.55µ g/100 ml. Plasma cortisol increased from 0.42 to 3.08µ g/100 ml during the 4 h ascent to 5,000 m and decreased to near normal levels the following day. A rhythmic plasma cortisol pattern was maintained after one day at simulated high altitude.     

Effects of adrenocorticotropin stimulation on cortisol dynamics of pregnant gilts and their fetuses: implications for prenatal stress studies

The effect of adrenocorticotropic hormone (ACTH) administration on plasma cortisol concentrations was determined in pregnant gilts and their fetuses. In a first experiment, 100 IU ACTH (Synacthen Depot) was administered intramuscularly to the gilts every second day from Days 49 to 75 of gestation. ACTH injections were carried out at 08:00 h and, thereafter, 10 blood samples were taken within the following 8h via jugular catheters. Blood samples were analysed for plasma cortisol concentrations, and results were compared with values from animals which were treated with physiological saline and untreated animals (blood sampling only). The values for plasma cortisol concentrations increased until 3h after ACTH applications to a mean maximum level of 276.5+/-17.2 nmol/l in the whole 4-week stimulation period. Plasma cortisol levels did not return to pre-treatment values within the 8 h post-injection. No differences in cortisol levels were found between the physiological saline and untreated control, and no habituation of the adrenocortical response to ACTH was found during the 4-week stimulation period. In a second experiment, 100 IU ACTH were administered to pregnant gilts at gestation Day 65. After 3 h, fetuses were recovered under general anaesthesia and blood samples were taken from the umbilical vein, artery, and, after decapitation, from periphery. Application of ACTH to the sows significantly increased their plasma cortisol concentrations (P<0.001), and also increased plasma cortisol concentrations in peripheral blood samples from the fetuses (P=0.09) and in the umbilical vein (P<0.001) and artery (P<0.01), respectively. Plasma ACTH concentrations did not differ in fetuses from ACTH-treated or control sows. The results show that in gilts the adrenocortical response to an exogenous application of Synacthen Depot is consistent over time during mid-gestation. Furthermore, cortisol but not ACTH levels were increased in fetuses from ACTH-treated sows, indicating that maternal cortisol can cross the placenta during mid-gestation. The stimulation of maternal cortisol release through exogenous ACTH with subsequent elevation of fetal cortisol levels is, therefore, a useful approach for studying effects of elevated maternal glucocorticoids in prenatal stress studies in pigs.     

Sustained but not repeated acute elevation of cortisol impaired the luteinizing hormone surge, estrus, and ovulation in gilts.

We tested the hypothesis that sustained and repeated acute elevation of cortisol would impair the LH surge, estrus, and ovulation in gilts. Cortisol was injected intramuscularly, to achieve a sustained elevation of plasma concentrations of cortisol, or intravenously, to achieve an acute elevation of plasma concentrations of cortisol. Control gilts received i.m. injections of oil and i.v. injections of saline. These treatments were administered to gilts (n = 6 per treatment) at 12-h intervals from Days 7 to 11 of the estrous cycle until after estrus ceased or until Day 27 or 28 of the estrous cycle, whichever came first. The repeated acute elevation of cortisol had no effect on the LH surge, estrus, or ovulation. In contrast, when the elevation of cortisol was sustained, the LH surge, estrus, and ovulation were inhibited. We conclude that cortisol is capable of direct actions to impair reproductive processes in female pigs but that plasma concentrations of cortisol need to be elevated for a substantial period for this to occur.     

Different pituitary beta-endorphin and adrenal cortisol response to ethanol in individuals with high and low risk for future development of alcoholism

The purpose of the present studies was to investigate the activity of the adrenal gland and the pituitary beta-endorphin system in individuals from families with a 3 generation history of alcoholism, High Risk group, or from families without history of alcoholism, Low Risk group. All subjects had a medical examination, a drinking behavior personal interview and the Michigan Alcoholism Screening Test. Individuals with medical problems or excessive drinking were not included in the study. On the day of testing, a blood sample was taken at 9:00 a.m., then the subject drank a placebo drink or an ethanol solution (0.5 g ethanol/kg B.Wt.). Additional blood samples were taken at 15, 45 and 120 minutes post-drink. Results indicated that individuals of the High Risk group had lower basal levels of beta-endorphin like immunoreactivity (beta-EPLIR) than individuals of the Low Risk group. The dose of 0.5 g ethanol/kg B.Wt. induced an increase in the plasma content of beta-EPLIR of the High Risk group, but not of the Low Risk group. In the Low Risk group ethanol did not induce an increase above the 9:00 a.m. levels, however, it attenuated the beta-endorphin decrease overtime, observed following the placebo drink. Analysis of beta-endorphin-like peptides in the plasma of the High Risk group, with Sephadex G-75 chromatography indicated that the major component of the plasma beta-EPLIR was beta-lipotropin. Plasma cortisol levels, following ethanol intake, presented a small increase in the High Risk group but not in the Low Risk group. Both groups presented similar blood alcohol levels. The basal levels of immunoreactive cortisol and beta-endorphin in the plasma of individuals who were alcoholics, but had been abstinent for at least six months prior to testing were similar to the levels of the High Risk group. Thus there are differences both in the basal levels and in the response of the cortisol and the pituitary beta-endorphin system to an acute ethanol challenge between the two groups.     

Effect of ACTH (tetracosactide) on steroid hormone levels in the mare. Part A: effect in intact normal mares and mares with possible estrous related behavioral abnormalities

Ovariectomized mares and mares with inactive ovaries may show signs of estrus. The reason behind this phenomenon is not clear; however, steroid hormones of adrenal origin have been suggested. Moreover, aberrant adrenal hormone production has been implied as a reason why some intact mares may change behavior. In the present study, the effect of ACTH on plasma levels of cortisol, progesterone, androstenedione and testosterone was investigated in intact mares with normal estrous behavior ('controls', n=5) and intact mares that according to their owners showed deviant estrous behavior ('problem' mares, n=7). Blood samples were collected hourly from 12:00 h until 14:00 h the following day (half-hourly between 14:00 and 17:00 h) on two occasions (at two estruses), with saline or ACTH treatment (tetracosactide) at 14:00 h (saline treatment day or ACTH treatment day). ACTH treatment caused a significant increase in plasma levels of cortisol, progesterone, androstenedione and testosterone in all mares (P<0.05). An overall significant difference in cortisol response to ACTH was found (P<0.05), with 'problem' mares showing a significantly lower increase in cortisol levels 30 min to 3h post ACTH treatment (P<0.001). The 'problem' mares also showed a significantly higher increase than controls in progesterone levels in the same time period (P<0.05). The reason for the reduced adreno-cortical reactivity, with a low cortisol response to the ACTH treatment, in the 'problem' mares is unknown, but may indicate a difference in adrenal function as compared to control mares.     

Effects of two different anaesthetics on serum concentrations of cortisol and luteinizing hormone in barrows and gilts

To determine if administration of the anaesthetic cocktail, telazol-ketamine-xylazine (TKX) and pentothal (PEN) decreases serum concentrations of luteinizing hormone (LH) in pigs, the following experiment was performed. On day 1, eight gilts and six barrows of similar weight (75 kg) were anaesthetized with TKX (1 mL/22.5 kg body weight [BW] intramuscularly) and indwelling jugular catheters were inserted. On days 2, 6 and 8 blood samples were taken every 20 min, for 4 h before pigs were administered saline (day 2) or anaesthetized with TKX (day 6) and PEN (8.9 mg/kg BW intravenously, day 8). Blood samples were taken every 20 min for 4 h following administration of saline and anaesthetics. Mean serum concentrations of LH and cortisol did not differ (P > 0.05) within barrows or gilts from before administration of saline (day 2) to following saline administration. Mean serum concentrations of LH and cortisol were not different (P > 0.05) within barrows and gilts before administration of TKX (pre-TKX) or PEN compared with day 2 samples. Following administration of TKX (post-TKX), mean serum concentrations of LH decreased (P < 0.05) and remained decreased for 140 min, while mean serum concentrations of cortisol increased (P < 0.05) post-TKX and remained elevated for 140 min. In gilts, mean serum concentrations of LH did not differ (P > 0.05) from pre- to post-TKX. However, mean serum concentrations of cortisol increased (P < 0.05) post-TKX in gilts and remained elevated for 240 min. Following administration of PEN, mean serum concentrations of LH and cortisol within barrows and gilts were not different (P > 0.05) from concentrations before administration of PEN. Administration of TKX to barrows and gilts increased serum concentrations of cortisol, but transient decreases in serum concentrations of LH were observed only in barrows, indicating gonadal status and/or sex may influence the ability of TKX to alter circulating concentrations of LH in pigs.     

Effectiveness of saliva collection and enzyme-immunoassay for the quantification of cortisol in socially housed baboons

Circulating cortisol levels are often used to assess the biological stress response in captive primates. Some methods commonly used to collect blood samples may alter the stress response. As such, noninvasive means to analyze cortisol levels are increasingly being developed. We adapted an existing collection method to simultaneously obtain saliva from multiple socially living hamadryas baboons (Papio hamadryas hamadryas) and validated an enzyme-immunoassay kit to quantify cortisol within the saliva samples. Over a period of 12 months, saliva samples were regularly collected from approximately half of the 18-member colony, representing younger monkeys who were more willing to participate. The assay met the four criteria typically used to assess the effectiveness of a new analytical technique: parallelism, precision, accuracy, and sensitivity. Cortisol levels were also proportional to those expected given published plasma levels of cortisol in baboons. Further, salivary cortisol levels increased in individuals following significant stress-related events, such as removal from the group, indicating biological validation. The technique provided a reliable and effective means to assess a physiological indicator of stress in a social group without initiating a stress response owing to handling or sedation, and provided a real-time assessment of cortisol levels and reactivity.