Response of fecal cortisol to stress in captive chimpanzees (Pan troglodytes)

This study examined whether fecal cortisol could be used as an index of stress responses. The stress responsiveness of fecal cortisol was tested with a stressor known to stimulate adrenal activity, the stress of anesthesia. Daily fecal and urine samples were collected from four captive chimpanzees (Pan troglodytes) before and after anesthetizations with Telazol^®/Ketasat^®. Tests of assay validity indicated that cortisol was measurable in chimpanzee fecal extracts. Fecal cortisol concentrations were significantly elevated 2 days after anesthetization, with elevations in seven of the eight treatments. The posttreatment peak was significantly greater than baseline values in three of the four subjects. Both fecal concentrations and proportionate increases in response to stress were significantly correlated with the corresponding values in urinary cortisol, confirming the stressfulness of these procedures and the stress responsiveness of fecal cortisol. These findings provide evidence for the application of fecal cortisol as a noninvasive index of physiologic stress in nonhuman primates. Am. J. Primatol. 44:57–69, 1998. © 1998 Wiley-Liss, Inc.     

Non‐invasive fecal hormone analysis and behavioral observations for monitoring stress responses in captive western lowland gorillas (Gorilla gorilla gorilla)

Non‐invasive techniques for monitoring the stress response in captive western lowland gorillas (Gorilla gorilla gorilla) were investigated. Fecal samples for cortisol measurement and concurrent behavioral data were collected from six individuals in a socially housed gorilla group (one adult male, three adult females and their three offspring) over a 7‐month period. Despite inter‐individual variation in the dynamics of fecal cortisol concentrations over time, several major secretory peaks coincided across individuals. High cortisol concentrations in feces were correlated with induced stressors or behavioral observations indicating high social tension, with a 1–2 day lag period. Entry progression order of the gorillas into a den complex and a supplant‐based dominance index were suitable indicators of overall dominance hierarchies, and fluctuations over time reflected periods of instability. Diurnal variation in fecal cortisol was not apparent when comparing afternoon and morning samples, however the sample collection interval was relatively short (3–5 hr). These results demonstrate the feasibility of monitoring stress responses based on the dynamics of both fecal cortisol excretion and behavior. This non‐invasive approach may be used for gauging responses to changes in husbandry, environment and group structure of captive gorillas. Zoo Biol 0:1–15, 2005. © 2005 Wiley‐Liss, Inc.     

Dextroamphetamine infusions in normals result in correlated increases of plasma beta-endorphin and cortisol immunoreactivity

Stress induces behavioral and hormonal alterations. In experimental and clinical studies, amphetamine administration may simulate or potentiate such effects. The endogenous opioid system has been implicated in the modulation of stress effects. Dextroamphetamine, 0.3 mg./kg. and placebo were infused on separate days in a double-blind cross over design to eight normal volunteers. At 30 minutes post infusion, plasma β-endorphin immunoreactivity was significantly (p < 0.05) increased. The increase ws significantly correlated (r = 0.76, p = 0.05) with volunteers' cortisol responses.     

Cortisol and β-endorphin levels in peripheral circulation around weaning in primiparous sows

Peripheral plasma patterns of cortisol and β-endorphin were compared around weaning in primiparous sows. Blood samples for hormone analysis were collected repeatedly on the day before weaning, on the day of weaning and on the day after weaning. Peak cortisol and β-endorphin levels occurred shortly after weaning on the day of weaning. Cortisol concentration on the day of weaning was greater (P < 0.001) than on the day before weaning, which in turn was higher (P < 0.05) than on the day after weaning. β-Endorphin concentrations on the day before weaning and on the day of weaning were similar, but were greater (P < 0.01) on both these days than on the day after weaning. Overall, cortisol and β-endorphin patterns followed a similar trend, which was more marked on the day before weaning and on the day of weaning than on the day after weaning. Pearson's correlation coefficients for peripheral plasma cortisol and β-endorphin secretion were 0.61 (P < 0.001) on the day before weaning, 0.63 (P < 0.001) on the day of weaning and 0.15 (P < 0.13) on the day after weaning. The present study indicates that suckling and weaning are stressful stimuli during which the peripheral plasma concentrations of cortisol and β-endorphin follow a similar trend.     

Narrow-band ultraviolet B (NB UV-B) exposures improve mood in healthy individuals differently depending on chronotype

ABSTRACT

The evening chronotype is associated with psychological symptoms such as depressed mood, while skin exposure to ultraviolet radiation (UVR) may affect mood and behavior through neural and humoral routes. This pilot study aimed to investigate the impact of whole-body narrow-band (NB) UV-B exposure on current mood state and circulating 25-hydroxyvitamin D3 (25(OH)D₃), interleukin-6 (IL-6), cortisol and β-endorphin (β-END) levels in healthy participants. Here, eleven healthy women received full-body NB UV-B exposures on four afternoons, and the chronotype was assessed with a shortened version of Horne and Östberg’s Morningness-Eveningness Questionnaire (MEQ). Perceived mood was evaluated using the Visual Analogue Scale (VAS), and serum 25(OH)D₃, IL-6, cortisol and β-END concentrations were monitored daily. Decreasing VAS values showed mood to improve significantly over the five days after the four suberythematous NB UV-B exposures (p = .038), and the more the circadian preference was inclined toward eveningness, the greater the improvement in the mood dimension of wellbeing (p = .021). Baseline mood state was correlated with baseline 25(OH)D₃ (r = ?0.54, 95% CI: ?0.86 to ?0.09) and with baseline cortisol (r = ?0.57, 95% CI: ?0.87 to ?0.04). During the NB UV-B exposures, 25(OH)D₃ increased significantly, as expected, and IL-6 declined significantly by ?0.35 (95% CI: ?0.69 to ?0.07) pg/mL from the initial values of 1.12 ± 0.66 pg/mL (p = .025). In conclusion, in our pilot study, NB UV-B exposure improved mood, especially among those with evening preference for their daily activities, as well as circulating 25(OH)D₃ levels, whereas circulating IL-6 levels decreased.

Abbreviations: UVR: Ultraviolet radiation; NB UV-B: narrow-band UV-B; VAS: Visual Analogue Scales; β-END: β-endorphin; IL-6: Interleukin-6     

Baseline,diurnal variations,and stress-induced changes of stress hormones in three captive beluga whales,Delphinapterus leucas

Concentrations of plasma adrenocorticotropic hormone (ACTH), cortisol, and aldosterone were investigated in three adult beluga whales (Delphinapterus leucas), held in a large outdoor public aquarium exhibit. The purpose of this study was to evaluate resting concentrations of these hormones and associated diurnal variations with routine interactions and medical procedures. Resting blood samples were collected voluntarily from the ventral fluke veins at predetermined times of the day to evaluate diurnal changes in analyte concentrations. In addition, hematology and serum chemistry analyses were performed to monitor health status and evaluate changes related to physical exam procedures. Analogous sampling was conducted during out-of-water physical examinations and before and after wading-contact sessions (WCS). Baseline stress hormone concentrations (± SD) were as follows: plasma ACTH (8.41 ± 5.8 pg/mL), serum cortisol (1.80 ± 0.71 g/dL), and serum aldosterone (11.42 ± 5.5 pg/mL). Plasma ACTH and cortisol concentrations were consistently higher in early morning than evening, while aldosterone was higher in the evening. All stress-related hormones were significantly elevated during physical examination. Plasma ACTH concentrations were most increased, 5–10-fold, during physical examination, whereas cortisol and aldosterone showed 2–4-fold elevations. Stress response analytes measured during the WCS did not differ significantly from baseline concentrations.     

Evaluation of physiological stress in free-ranging bears: current knowledge and future directions

Stress responses, which are mediated by the neurogenic system (NS) and hypothalamic–pituitary–adrenal (HPA) axis help vertebrates maintain physiological homeostasis. Fight-or-flight responses are activated by the NS, which releases norepinephrine/noradrenaline and epinephrine/adrenaline in response to immediate stressors, whilst the HPA axis releases glucocorticoid hormones (e.g. cortisol and corticosterone) to help mitigate allostatic load. There have been many studies on stress responses of captive animals, but they are not truly reflective of typical ranges or the types of stressors encountered by free-ranging wildlife, such as responses and adaptation to environmental change, which are particularly important from a conservation perspective. As stress can influence the composition of age and sex classes of free-ranging populations both directly and indirectly, ecological research must be prioritised towards more vulnerable taxa. Generally, large predators tend to be particularly at risk of anthropogenically driven population declines because they exhibit reduced behavioural plasticity required to adapt to changing landscapes and exist in reduced geographic ranges, have small population sizes, low fecundity rates, large spatial requirements and occupy high trophic positions. As a keystone species with a long history of coexistence with humans in highly anthropogenic landscapes, there has been growing concern about how humans influence bear behaviour and physiology, via numerous short- and long-term stressors. In this review, we synthesise research on the stress response in free-ranging bear populations and evaluate the effectiveness and limitations of current methodology in measuring stress in bears to identify the most effective metrics for future research. Particularly, we integrate research that utilised haematological variables, cardiac monitors and Global Positioning System (GPS) collars, serum/plasma and faecal glucocorticoid concentrations, hair cortisol levels, and morphological metrics (primarily skulls) to investigate the stress response in ursids in both short- and long-term contexts. We found that in free-ranging bears, food availability and consumption have the greatest influence on individual stress, with mixed responses to anthropogenic influences. Effects of sex and age on stress are also mixed, likely attributable to inconsistent methods. We recommend that methodology across all stress indicators used in free-ranging bears should be standardised to improve interpretation of results and that a wider range of species should be incorporated in future studies.     

Antibodies Specific for α-N-Acetyl-β-Endorphins: Radioimmunoassays and Detection of Acetylated β-Endorphins in Pituitary Extracts

Abstract: Antibodies specific for α-N-acetyl-β-endorphins have been prepared by injecting into rabbits either α-N-acetyl-β-endorphin(1-31) or [α-N-acetyl, ε-acetyl-Lys⁹]-β-endorphin(1-9) linked by carbodiimide to bovine thyroglobulin. Both antisera were used to develop specific radioimmunoassays for α-N-acetyl-β-endorphins. The radioimmunoassays were used to measure α-N-acetylated β-endorphins in extracts of pituitary regions from different species. By comparison of the amounts of total β-endorphin and α-N-acetyl-β-endorphin immunoreactivity, a relative ratio of β-endorphin acetylation was obtained. The relative acetylation of β-endorphin was highest in rat posterior-intermediate lobe extracts (>90%). Beef and monkey intermediate lobes had a lower degree of acetylation (53 and 31%, respectively). Anterior lobe extracts from all three species contained low amounts of acetylated β-endorphin. Human pituitary extracts did not contain acetylated β-endorphins. By the use of cation exchange and high performance liquid chromatography, six different acetylated derivatives and fragments of β-endorphin were resolved in extracts of rat posterior-intermediate pituitaries. Two of these peptides corresponded to α-N-acetyl-β-endorphin(1-31) and -(1-27). One acetylated β-endorphin fragment had the same size as α-N-acetyl-β-endorphin(1-27) but was eluted earlier from the cation exchange column. This peptide had full cross-reactivity with antibodies directed against the middle and amino-terminal parts of β-endorphin. Compared with α-N-acetyl-β-endorphin(1-27), it had much less cross-reactivity with antibodies directed against the COOH-terminal part of β-endorphin, suggesting that it was a COOH-terminally modified derivative of β-endorphin(1-27). The remaining N-acetylated β-endorphin derivatives were eluted even earlier from the cation exchange column. The majority of these fragments were slightly larger in size than y-endorphin, i.e., β-endorphin(1-17), but smaller than β-endorphin(1-27). They had full cross-reactivity in an amino-terminally directed β-endorphin radioimmunoassay and a greatly diminished cross-reactivity with antibodies to the middle region of β-endorphin.     

Experimental Verification of the Effects on Normal Domestic Cats by Feeding Prescription Diet for Decreasing Stress

The objective of this study was to evaluate the effects of diet on the feline stress response by measuring plasma and urinary cortisol. A study diet was developed with a unique combination of nutrients that supports the management of stressful situations. The specific formulation of the diet included alpha-casozepine, which is believed to have an anxiolytic effect, and tryptophan supplementation. Tryptophan is the precursor for the synthesis of the neurotransmitter serotonin. Twenty-one indoor cats were fed with the study diet (n = 10) or a control diet (n = 11) for 8 weeks, after which physiological responses were evaluated. The study diet significantly increased the ratio of plasma tryptophan to large neutral amino acids and decreased urinary cortisol concentrations after being consumed daily for 8 weeks, but there was no effect on plasma cortisol levels following a stressful event (veterinary examination and blood draw). Further studies, such as behavioral analyses, are needed to clarify the effects of the study diet.     

Physiologic responses of grizzly bears to different methods of capture

The physiologic effects of two methods of capture, chemical immobilization of free-ranging (FR) bears by remote injection from a helicopter and physical restraint (PR) by leg-hold snare prior to chemical immobilization, were compared in 46 grizzly bears (Ursus arctos) handled during 90 captures between 1999 and 2001. Induction dosages and times were greater for FR bears than PR bears, a finding consistent with depletion of, or decreased sensitivity to, catecholamines. Free-ranging bears also had higher rectal temperatures 15 min following immobilization and temperatures throughout handling that correlated positively with induction time. Physically restrained bears had higher white blood cell counts, with more neutrophils and fewer lymphocytes and eosinophils, than did FR bears. This white blood cell profile was consistent with a stress leukogram, possibly affected by elevated levels of serum cortisol. Serum concentrations of alanine aminotransferase, aspartate aminotransferase, and creatine kinase were higher in PR bears that suggested muscle injury. Serum concentrations of sodium and chloride also were higher in PR bears and attributed to reduced body water volume through water deprivation and increased insensible water loss. Overall, different methods of capture resulted in different patterns of physiologic disturbance. Reducing pursuit and drug induction times should help to minimize increase in body temperature and alteration of acid-base balance in bears immobilized by remote injection. Minimizing restraint time and ensuring snare-anchoring cables are short should help to minimize loss of body water and prevent serious muscle injury in bears captured by leg-hold snare.     

Presence of immunoreactive alpha-endorphin in rat pituitary gland.

Utilizing radioimmunoassay for α-endorphin, we attempted to demonstrate immunoreactive α-endorphin in acid extracts of pars distalis and combined pars intermedia and pars nervosa of the rat pituitary gland after chromatography on Sephadex G-25. β-Lipotropin, β-endorphin and γ-endorphin were not converted into α-endorphin during the extraction and gel chromatographic procedures. Concentrations of immunoreactive α-endorphin determined after gel chromatography of extracts from pars distalis and combined pars intermedia and pars nervosa were 1.1±0.6 and 130±17 ng/mg wet tissue (mean±SE), respectively. Serial dilution of these extracts gave parallel lines to the standard curve of synthetic α-endorphin, but not to that of γ-endorphin or δ-endorphin. These results suggest the existence of immunoreactive α-endorphin indistinguishable in molecular size from synthetic α-endorphin in the rat pituitary gland.     

Alterations in blood and urine parameters in two florida manatees (Trichechus manatus latirostris) from simulated conditions of release following rehabilitation

In an effort to manage the existing population of the endangered Florida manatee (Trichechus manatus latirostris), as many individuals as possible are rehabilitated from illness or injury and released back into the waters of the state of Florida. It is not uncommon, however, for manatees recaptured for health assessment following release from rehabilitation to have elevated concentrations of serum creatinine. Although such elevated levels would be indicative of kidney failure in most other mammals, problems associated with renal function have not been evident in these recaptured manatees. To determine the possible cause(s) of the serum creatinine increase, two captive Florida manatees were manipulated to simulate many of the environmental and physical changes that occur during and shortly after release. Routine chemical analyses of serum and urine, complete blood counts, serum cortisol concentrations, and lymphocyte proliferation responses were measured. Serum creatinine concentrations increased significantly in response to decreased food intake and changes in food type. The increases differed depending on the salinity of the water in which the animals were maintained. It was found that significant changes in urinary creatinine and serum creatine kinase occurred as well, but serum cortisol concentrations were elevated only during simulated transport. Lymphocyte proliferation assays indicated that immune function was potentially impaired by extreme levels of dietary restriction and by changes in salinity. These results suggest that serum creatinine elevations and subsequent effects on the immune system might be minimized by adapting manatees undergoing rehabilitation to the diet and salinity they would encounter following release. Zoo Biol 22:103–120, 2003. © 2003 Wiley‐Liss, Inc.     

Diurnal variation of salivary cortisol in captive African elephants (Loxodonta africana) under routine management conditions and in relation to a translocation event

The present study assessed the diurnal variation in salivary cortisol in captive African elephants during routine management (baseline) and in relation to a potential stressor (translocation) to evaluate to what extent acute stress may affect diurnal cortisol patterns. Under baseline conditions, we collected morning and afternoon saliva samples of 10 animals (three zoos) on different days in two study periods (n = 3–10 per animal, daytime and period). Under stress conditions, we sampled the transported cow (newcomer) and the two cows of the destination zoo before and after the transport in the morning and afternoon (n = 3–9 per animal, daytime and transport phase), as well as after the first introduction of the newcomer to the bull (n = 1 per animal). Cortisol was measured in unextracted samples by enzyme immunoassay. Under baseline conditions, we observed the expected diurnal variation with higher cortisol levels in the morning than in the afternoon. Under stress conditions, neither a significant difference between pre- and posttransport, nor between morning and afternoon levels was found. The percentage difference between morning and afternoon cortisol after the transport, however, was remarkably lower than before the transport in the newcomer potentially indicating a stress response to familiarization. Saliva samples taken immediately after the introduction of the newcomer to the bull revealed a marked cortisol increase. Our findings indicate that stressors may disturb the diurnal cortisol rhythm. Furthermore, provided that samples can be collected promptly, salivary cortisol is a useful minimally invasive measure of physiological stress in the African elephant.     

Polar bear stress hormone cortisol fluctuates with the North Atlantic Oscillation climate index

Polar bears are heavily dependent on sea ice for hunting sufficient prey to meet their energetic needs. When the bears are left fasting, it may cause a rise in the levels of the stress hormone cortisol. Cortisol is the major corticosteroid hormone in most mammals, including polar bears. Production and regulation of this stress hormone are vital for the body as it is part of a myriad of processes, including in relation to metabolism, growth, development, reproduction, and immune function. In the present study, we examined the correlation between East Greenland polar bear hair cortisol concentration (HCC), a matrix that reflects longer-term hormone levels, and the fluctuations of the North Atlantic Oscillation (NAO) index, a large-scale climate phenomenon applied as a proxy for sea ice extent in the Greenland Sea along the coast of East Greenland. In doing so, a significant positive correlation (r = 0.88; p = 0.0004) was found between polar bear hair cortisol and the NAO, explaining 77 % of the variation in HCC observed between years over the period 1989–2009. This result indicates that interannual fluctuations in climate and ice cover have a substantial influence on longer-term cortisol levels in East Greenland polar bears. Further research into the implications and consequences inherent in this correlation are recommended, preferably across multiple polar bear populations.     

Serum corticosteroid binding globulin expression is modulated by fasting in polar bears (Ursus maritimus)

Polar bears (Ursus maritimus) from several subpopulations undergo extended fasting during the ice-free season. However, the animals appear to conserve protein despite the prolonged fasting, though the mechanisms involved are poorly understood. We hypothesized that elevated concentrations of corticosteroid binding globulin (CBG), the primary cortisol binding protein in circulation, lead to cortisol resistance and provide a mechanism for protein conservation during extended fasting. The metabolic state (feeding vs. fasting) of 16 field sampled male polar bears was determined based on their serum urea to creatinine ratio (>25 for feeding vs. <5 for fasting). There were no significant differences in serum cortisol levels between all male and female polar bears sampled. Serum CBG expression was greater in lactating females relative to non-lactating females and males. CBG expression was significantly higher in fasting males when compared to non-fasting males. This leads us to suggest that CBG expression may serve as a mechanism to conserve protein during extended fasting in polar bears by reducing systemic free cortisol concentrations. This was further supported by a lower serum glucose concentration in the fasting bears. As well, a lack of an enhanced adrenocortical response to acute capture stress supports our hypothesis that chronic hunger is not a stressor in this species. Overall, our results suggest that elevated serum CBG expression may be an important adaptation to spare proteins by limiting cortisol bioavailability during extended fasting in polar bears.     

The efficacy of orally dosed ketamine and ketamine/medetomidine compared with intramuscular ketamine in rhesus macaques (Macaca mulatta) and the effects of dosing route on haematological stress markers

BACKGROUND: This study compared the efficacy of two orally-dosed (PO) anaesthetic regimens for chemical immobilization in rhesus macaques (Macaca mulatta), versus the standard protocol of intramuscular (TM) ketamine. In addition, the effects of dosing route on haematological stress markers were evaluated. METHODS: Testing was conducted on 18 chronically housed animals. Animals were trained to accept oral dosing and then randomly assigned to one of three drug regimens: (1) ketamine IM, (2) ketamine PO, (3) Ketamine/medetomidine PO. Sedation levels for each regimen were evaluated. RESULTS: Oral dosing alone was not sufficient to achieve a plane of sedation that allowed for safe handling. Serum cortisol and glucose levels were unchanged across groups, although differences were observed in the leukogram profiles. CONCLUSION: The oral dosages used in this study fell short in providing adequate sedation for safe handling for routine veterinary procedures. Leukogram profiles indicated that orally dosed animals experienced a higher level of stress.     

Social relationship and hair cortisol level in captive male chimpanzees (<Emphasis Type="Italic">Pan troglodytes</Emphasis>)

Understanding how social relationships affect long-term stress is important because stress has a profound impact on the welfare of animals and social relationships often exert a strong influence on their stress responses. The purpose of this study was to investigate the relationship between social behaviors and long-term stress levels as assessed by hair cortisol (HC) concentration. The subjects were 11 chimpanzees living in an all-male group (divided into two sub-groups) in Kumamoto Sanctuary, Kyoto University, Japan. Behavioral data were collected between December 2014 and March 2015. The total observation time was 129 h. Hair samples were collected in late March and early April 2015, and cortisol was extracted from the hair and measured with enzyme immunoassay. The hair growth rate was estimated to be 1.33 ± 0.06 cm/month. The results revealed that there was a positive correlation between the rate of receiving aggression and HC levels. We also found a significant negative correlation between the balance between giving and receiving grooming (grooming balance index: GBI), which was calculated by subtracting the rate with which grooming is given from that with which it is received, and the rate of receiving aggression and between the GBI and HC levels. Thus, individuals receiving higher levels of aggression also tended to give grooming for relatively long periods compared to the time they were being groomed. In contrast, the rate of initiating aggression did not have a relationship with either HC levels or any measure of social grooming. We did not find social buffering effects, as there was no correlation between mutual social grooming and HC levels. These results show that not only aggressive interactions but also overall social situations in which animals do not have balanced relationships with others might result in the long-term elevation of cortisol levels in captive male chimpanzees.     

Serum parameters of Adriatic sturgeon Acipenser naccarii (Pisces: Acipenseriformes): effects of temperature and stress

Data on the concentrations of some blood constituents of captive Adriatic sturgeon, Acipenser naccarii, a primitive bony fish, are reported. Serum osmolality, Na⁺, K⁺, Cl⁻, Ca²⁺, cortisol, glucose and total protein concentrations were measured. The effects of anaesthesia, temperature, crowding and prolonged handling stress were tested on a group of 12 4-year-old sturgeons sampled repeatedly. The anaesthetic dose of MS 222 (140 mg l⁻¹) induced significant osmolality elevation in the sturgeon. After exposure to colder temperature (17 versus 25°C), cortisol and Cl⁻ concentrations significantly decreased. The cultured sturgeon did not seem susceptible to crowding and prolonged handling stress, since neither the serum cortisol and glucose levels nor the other blood parameters were affected by these stressors. Results are compared with the few available data on other chondrostean fish and with those on teleosts.     

Acute Thermal and Stress Response in Moose to Chemical Immobilization

Management and research of moose (Alces alces) in Alaska, USA, often require chemical immobilization; however, moose may be prone to capture-induced hyperthermia while immobilized. We chemically immobilized moose with carfentanil citrate and xylazine hydrochloride to measure rump fat depth, collect blood and fecal samples, and to deploy modified vaginal implant transmitters and global positioning system (GPS)-collars for recording body temperature and movement during and after the chemical immobilization. We predicted wild moose pursued and captured from a helicopter would have elevated body temperature at time of capture, whereas body temperature would remain stable in hand-raised captive moose not pursued and only hand-injected for immobilization. Additionally, we expected post-capture body temperature would be a function of activity, time immobilized, and ambient temperature. As predicted, body temperature of wild moose was elevated 1 hour after capture (38.9°C, 95% CI = 38.7–39.1°C) but returned to baseline levels within 3 hours (38.0°C, 95% CI = 37.9–38.1°C); however, body temperatures then rose above baseline levels and remained elevated 12–48 hours post-capture when movement rates were also elevated. Body temperatures in captive moose were not elevated 1-hour post-immobilization (37.9°C, 95% CI = 37.8–38.0°C). Body temperatures of wild moose were positively related to cortisol levels at time of capture. Two moose that died after immobilization had initial body temperatures similar to other immobilized moose; however, their body temperature began to rise at 17 hours and 40 hours post-immobilization. Our study provides evidence that chemical immobilization affects body temperature and movement of wild moose up to 48 hours after capture, possibly as a result of renarcotization from carfentanil citrate. With advancements in technology, we recommend fine-scale GPS data (<1-hr fix rates) and continuous body temperature be evaluated to detect evidence of renarcotization during and after opioid-based captures of northern ungulates. © 2020 The Wildlife Society.     

α-N-Acetyl β-Endorphins in the Pituitary: Immunohistochemical Localization Using Antibodies Raised Against Dynorphin(1-13)

Abstract: Intense immunohistochemical staining of the intermediate lobe of the pituitary was observed by using an antiserum raised against synthetic dynorphin(1-13) treated with a water-soluble carbodiimide (CDI). Subsequent studies showed that the immunostaining was blocked by preincubation of the antiserum with acetylated derivatives of both β-endorphin and dynorphin(1-13) as well as by CDI-treated dynorphin(1-13), but only weakly by authentic dynorphin(1-13). Neither nonacetylated β-endorphin nor any other fragments of the ACTH/endorphin precursor blocked the immunostaining of the intermediate lobe. Analysis of the CDI-treated dynorphin(1-13) used as an antigen showed that most of the peptide was acetylated at primary amino groups. CDI treatment of dynorphin(1-13) results in the formation of an acetyl derivative because the commercially available peptide is supplied as the acetate salt. The antibodies responsible for the intermediate lobe staining were isolated by affinity chromatography, using a column containing partially purified intermediate lobe extract linked to an affinity resin and a radioimmunoassay (RIA) was developed with CDI-treated dynorphin(1-13) used as a trace and as a standard. Competition studies showed 0.5-1% cross-reactivity with α-N-acetyl β-endorphin(1-31), α-N-acetyl β-endorphin(1-27), and totally acetylated β-endorphin(1-31). Nonacetylated β-endorphins did not cross-react. Posterior-intermediate lobe extracts from rat and beef were fractionated by gel filtration. Rat posterior-intermediate lobe extracts were also fractionated by cation-exchange chromatography. Fractionated extracts were analyzed by RIAs for β-endorphin, CDI-treated dynorphin(1-13), and authentic dynorphin(1-13). The results suggested that the peptides responsible for the intermediate lobe staining were mainly four different derivatives of β-endorphin bearing an acetyl group at the amino terminus. No immunostaining was seen in the posterior and anterior lobes of the pituitary. This suggests that the intermediate lobe is the main source of acetylated β-endorphins in the pituitary.