Ligand-Free Signaling of G-Protein-Coupled Receptors: Relevance to µ Opioid Receptors in Analgesia and Addiction.

Numerous G-protein-coupled receptors (GPCRs) display ligand-free basal signaling with potential physiological functions, a target in drug development. As an example, the µ opioid receptor (MOR) signals in ligand-free form (MOR-µ*), influencing opioid responses. In addition, agonists bind to MOR but can dissociate upon MOR activation, with ligand-free MOR-µ* carrying out signaling. Opioid pain therapy is effective but incurs adverse effects (ADRs) and risk of opioid use disorder (OUD). Sustained opioid agonist exposure increases persistent basal MOR-µ* activity, which could be a driving force for OUD and ADRs. Antagonists competitively prevent resting MOR (MOR-µ) activation to MOR-µ*, while common antagonists, such as naloxone and naltrexone, also bind to and block ligand-free MOR-µ*, acting as potent inverse agonists. A neutral antagonist, 6ß-naltrexol (6BN), binds to but does not block MOR-µ*, preventing MOR-µ activation only competitively with reduced potency. We hypothesize that 6BN gradually accelerates MOR-µ* reversal to resting-state MOR-µ. Thus, 6BN potently prevents opioid dependence in rodents, at doses well below those blocking antinociception or causing withdrawal. Acting as a 'retrograde addiction modulator', 6BN could represent a novel class of therapeutics for OUD. Further studies need to address regulation of MOR-µ* and, more broadly, the physiological and pharmacological significance of ligand-free signaling in GPCRs.

Comparison of some cardiopulmonary effects of etorphine and thiafentanil during the chemical immobilization of blesbok (Damaliscus pygargus phillipsi).

OBJECTIVE: To determine the cardiopulmonary effects of etorphine and thiafentanil for immobilization of blesbok. STUDY DESIGN: Blinded, randomized, two-way crossover study. ANIMALS: A group of eight adult female blesbok. METHODS: Animals were immobilized twice, once with etorphine (0.09 mg kg-1) and once with thiafentanil (0.09 mg kg-1) administered intramuscularly by dart. Immobilization quality was assessed and analysed by Wilcoxon signed-rank test. Time to final recumbency was compared between treatments by one-way analysis of variance. Cardiopulmonary effects including respiratory rate (ƒR), arterial blood pressures and arterial blood gases were measured. A linear mixed model was used to assess the effects of drug treatments over the 40 minute immobilization period. Significant differences between treatments, for treatment over time as well as effect of treatment by time on the variables, were analysed (p < 0.05). RESULTS: There was no statistical difference (p = 0.186) between treatments for time to recumbency. The mean ƒR was lower with etorphine (14 breaths minute-1) than with thiafentanil (19 breaths minute-1, p = 0.034). The overall mean PaCO2 was higher with etorphine [45 mmHg (6.0 kPa)] than with thiafentanil [41 mmHg (5.5 kPa), p = 0.025], whereas PaO2 was lower with etorphine [53 mmHg (7.1 kPa)] than with thiafentanil [64 mmHg (8.5 kPa), p < 0.001]. The systolic arterial pressure measured throughout all time points was higher with thiafentanil than with etorphine (p = 0.04). The difference varied from 30 mmHg at 20 minutes after recumbency to 14 mmHg (standard error difference 2.7 mmHg) at 40 minutes after recumbency. Mean and diastolic arterial pressures were significantly higher with thiafentanil at 20 and 25 minute measurement points only (p < 0.001). CONCLUSIONS: Both drugs caused clinically relevant hypoxaemia; however, it was less severe with thiafentanil. Ventilation was adequate. Hypertension was greater and immobilization scores were lower with thiafentanil.

Comparison of cardiopulmonary effects of etorphine and thiafentanil administered as sole agents for immobilization of impala (Aepyceros melampus).

OBJECTIVE: To compare the cardiopulmonary effects of the opioids etorphine and thiafentanil for immobilization of impala. STUDY DESIGN: Two-way crossover, randomized study. ANIMALS: A group of eight adult female impala. METHODS: Impala were given two treatments: 0.09 mg kg-1 etorphine or 0.09 mg kg-1 thiafentanil via remote dart injection. Time to recumbency, quality of immobilization and recovery were assessed. Respiratory rate, heart rate (HR), mean arterial blood pressure (MAP) and arterial blood gases were measured. A linear mixed model was used to analyse the effects of treatments, treatments over time and interactions of treatment and time (p < 0.05). RESULTS: Time to recumbency was significantly faster with thiafentanil (2.0 ± 0.8 minutes) than with etorphine (3.9 ± 1.6 minutes; p = 0.007). Both treatments produced bradypnoea, which was more severe at 5 minutes with thiafentanil (7 ± 4 breaths minute-1) than with etorphine (13 ± 12 breaths minute-1; p = 0.004). HR increased with both treatments but significantly decreased over time when etorphine (132 ± 17 to 82 ± 11 beats minute-1) was compared with thiafentanil (113 ± 22 to 107 ± 36 beats minute-1; p < 0.001). Both treatments caused hypertension which was more profound with thiafentanil (mean overall MAP = 140 ± 14 mmHg; p < 0.001). Hypoxaemia occurred with both treatments but was greater with thiafentanil [PaO2 37 ± 13 mmHg (4.9 kPa)] than with etorphine [45 ± 16 mmHg (6.0 kPa)] 5 minutes after recumbency (p < 0.001). After 30 minutes, PaO2 increased to 59 ± 10 mmHg (7.9 kPa) with both treatments (p < 0.001). CONCLUSIONS AND CLINICAL RELEVANCE: The shorter time to recumbency with thiafentanil may allow easier and faster retrieval in the field. However, thiafentanil caused greater hypertension, and ventilatory effects during the first 10 minutes, after administration.

Immobilization quality and cardiopulmonary effects of etorphine alone compared with etorphine-azaperone in blesbok (Damaliscus pygargus phillipsi).

OBJECTIVE: To evaluate the immobilization quality and cardiopulmonary effects of etorphine alone compared with etorphine-azaperone in blesbok (Damaliscus pygargus phillipsi). STUDY DESIGN: Blinded, randomized, crossover design. ANIMALS: A total of 12 boma-habituated female blesbok weighing [mean ± standard deviation (SD)] 57.5 ± 2.5 kg. METHODS: Each animal was administered etorphine (0.09 mg kg-1) or etorphine-azaperone (0.09 mg kg-1; 0.35 mg kg-1) intramuscularly with 1-week intertreatment washout period. Time to first sign of altered state of consciousness and immobilization time were recorded. Physiological variables were recorded, arterial blood samples were taken during a 40-minute immobilization period, and naltrexone (mean ± SD: 1.83 ± 0.06 mg kg-1) was intravenously administered. Recovery times were documented, and induction, immobilization and recovery were subjectively scored. Statistical analyses were performed; p < 0.05 was significant. RESULTS: No difference was observed in time to first sign, immobilization time and recovery times between treatments. Time to head up was longer with etorphine-azaperone (0.5 ± 0.2 versus 0.4 ± 0.2 minutes; p = 0.015). Etorphine caused higher arterial blood pressures (mean: 131 ± 17 versus 110 ± 11 mmHg, p < 0.0001), pH, rectal temperature and arterial oxygen partial pressure (59.2 ± 7.7 versus 42.2 ± 9.8 mmHg), but lower heart (p = 0.002) and respiratory rates (p = 0.01). Etorphine-azaperone combination led to greater impairment of ventilatory function, with higher end-tidal carbon dioxide (p < 0.0001) and arterial partial pressure of carbon dioxide (58.0 ± 4.5 versus 48.1 ± 5.1 mmHg). Immobilization quality was greater with etorphine-azaperone than with etorphine alone (median scores: 4 versus 3; p < 0.0001). CONCLUSIONS AND CLINICAL RELEVANCE: Both treatments provided satisfactory immobilization of blesbok; however, in addition to a deeper level of immobilization, etorphine-azaperone caused greater ventilatory impairment. Oxygen supplementation is recommended with both treatments.

EVALUATION OF THE QUALITY OF IMMOBILIZATION AND CARDIORESPIRATORY EFFECTS OF ETORPHINE-MEDETOMIDINE-AZAPERONE COMBINATION IN PLAINS ZEBRAS (EQUUS QUAGGA): A PILOT STUDY.

Five free-ranging male (subadults, n = 3; adults, n = 2) plains zebras (Equus quagga) were immobilized using a combination of etorphine (0.017 mg/kg), medetomidine (0.017 mg/kg), and azaperone (0.24 mg/kg) by means of a blank cartridge-fired projector. Time to recumbency was recorded and a descriptive score used to assess the quality of immobilization, manipulation, maintenance, and recovery. Physiological parameters were recorded at 5-min intervals for 20 min. At the end of the procedure, naltrexone (0.23 mg/kg) was administered intramuscularly and time to standing documented. The combination evaluated in this study allowed for successful immobilization and safe recovery of all animals, including during the subsequent 15 days. Despite the good outcome in this pilot study, as a result of the periodic apneic events and hypercapnia documented in the zebras, the authors suggest that physiological parameters be thoroughly monitored when using this protocol. Further studies are needed to improve upon chemical immobilization protocols in free-ranging plains zebras.

Dose-effect study of the serotonin agonist R-8-OH-DPAT on opioid-induced respiratory depression in blesbok (Damaliscus pygargus philipsi) and impala (Aepyceros melampus).

OBJECTIVE: To determine whether the R-enantiomer of 8-hydroxy-2-(di-n-propylamino) tetralin (R-8-OH-DPAT) alleviates respiratory depression in antelope species immobilized with etorphine. The experiment also aimed to establish the most clinically effective dose of this serotonin 5- HT1A receptor agonist. ANIMALS: A group of six female blesbok and six female impala. STUDY DESIGN: Each animal was subjected to four immobilization treatments in a prospective four-way crossover design-control treatment consisting of only etorphine at 0.09 mg kg-1 and three treatments consisting of etorphine at 0.09 mg kg-1 combined with 0.005, 0.02 and 0.07 mg kg-1 of R-8-OH-DPAT, respectively. Induction, quality of immobilization and recovery were monitored in each treatment. Physiological variables including heart rate, respiratory rate, arterial blood pressure and blood gases were measured for 35 minutes during immobilization. A linear mixed model was used to assess the effects of treatments over the recumbency period. RESULTS: R-8-OH-DPAT did not influence induction, immobilization or recovery scores. Respiratory rate in blesbok was increased in the medium- and high-dosage R-8-OH-DPAT treatment group. However, this increased respiratory rate did not translate into improvements of arterial partial pressure of oxygen (PaO2) values in the blesbok. The medium and higher dosages of R-8-OH-DPAT in impala led to an improved PaO2 as well as to decreased opioid-induced tachycardia during the first 10 minutes of immobilization. CONCLUSIONS AND CLINICAL RELEVANCE: Previous reports indicated that the racemic mixture of 8-OH-DPAT injected intravenously had a positive effect on blood-gas values in etorphine-treated hypoxemic goats. In this experiment, similar effects could be seen in impala at the higher dosage rates of R-8-OH-DPAT. However, failure to achieve an improvement of blood-gas values in blesbok was an unexpected result. It could be speculated that the dosage, species-specific differences of serotonin receptors or the use of the R-enantiomer of 8-OH-DPAT might play a role.

A COMPARISON OF OPIOID-BASED PROTOCOLS FOR IMMOBILIZATION OF CAPTIVE GREVY'S ZEBRA (EQUUS GREVYI).

Grevy's zebra (Equus grevyi) is an endangered species often found in zoological collections. Veterinary care for this species often requires immobilization. This study retrospectively evaluated behavioral and physiological parameters from Grevy's zebra records after three immobilization protocols: etorphine and alpha-2 agonist (EA2; n = 11); etorphine, alpha-2 agonist, and ketamine (EA2K; n = 16); and thiafentanil, alpha-2 agonist, and ketamine (TA2K; n = 6). Median time to working depth was statistically different (P = 0.03; EA2 = 6.5 min, EA2K = 6.3 min, TA2K = 14.5 min) by the Kruskal-Wallis test. When EA2 + EA2K were combined and compared with TA2K (Wilcoxon rank sum test), median recumbency time (P = 0.02) was also significantly longer (15 min compared with 6 min) for TA2K. There were no significant differences between the groups for physiological parameters or recovery time after reversal. Although all protocols produced anesthesia in Grevy's zebra, increased time to achieve working depth was observed with the alpha-2 and thiafentanil combinations.

Postinduction butorphanol administration alters oxygen consumption to improve blood gases in etorphine-immobilized white rhinoceros.

OBJECTIVE: To investigate the effects of postinduction butorphanol administration in etorphine-immobilized white rhinoceros on respiration and blood gases. STUDY DESIGN: Randomized crossover study. ANIMALS: A group of six sub-adult male white rhinoceros. METHODS: Etorphine, or etorphine followed by butorphanol 12 minutes after recumbency, was administered intramuscularly [2.5 mg etorphine, 25 mg butorphanol (1000-1250 kg), or 3.0 mg etorphine, 30 mg butorphanol (1250-1500 kg)]. Sampling started at 10 minutes after initial recumbency, and was repeated at 5 minute intervals for 25 minutes. Arterial blood gases, limb muscle tremors, expired minute ventilation and respiratory frequency were measured at each sampling point. Calculated values included alveolar-arterial oxygen gradient [ [Formula: see text] ], expected respiratory minute volume (V˙e), tidal volume (Vt), oxygen consumption ( [Formula: see text] ) and carbon dioxide production ( [Formula: see text] ). RESULTS: Etorphine administration resulted in an initial median (range) hypoxaemia [arterial partial pressure of oxygen 25.0 (23.0-28.0) mmHg], hypercapnia [arterial partial pressure of carbon dioxide 76.2 (67.2-81.2) mmHg], increased [Formula: see text] [41.7 (36.6-45.1) mmHg, [Formula: see text] [11.1 (10.0-12.0) L minute-1] and muscle tremors. Butorphanol administration was followed by rapid, although moderate, improvements in arterial partial pressure of oxygen [48.5 (42.0-51.0) mmHg] and arterial partial pressure of carbon dioxide [62.8 (57.9-75.2) mmHg]. In rhinoceros administered butorphanol, [Formula: see text] [4.4 (3.6-5.1) L minute-1] and [Formula: see text] [4.2 (3.8-4.4) L minute-1] were lower than in those not administered butorphanol. Increased arterial oxygen tension was associated with lower oxygen consumption (p=0.002) which was positively associated with lower muscle tremor scores (p<0.0001). CONCLUSIONS AND CLINICAL RELEVANCE: Hypoxaemia and hypercapnia in etorphine-immobilized rhinoceros resulted from an increased [ [Formula: see text] ] and increased [Formula: see text] and [Formula: see text] associated with muscle tremors. Rather than being associated with changes in V˙e, it appears that improved blood gases following butorphanol administration were a consequence of decreased [Formula: see text] associated with reduced muscle tremoring.

IMMOBILIZING MUSKOX ( OVIBOS MOSCHATUS) UNDER HIGH ARCTIC CONDITIONS.

Immobilizing and handling large, free-ranging animals without proper facilities in harsh environmental conditions poses significant challenges. During two field expeditions, a total of 29 female muskoxen ( Ovibos moschatus) were immobilized in Northeast Greenland (74°N, 20°E). Fixed doses of immobilizing drugs were used regardless of animal size: 2 mg etorphine, 30 mg xylazine, 0.3 mg medetomidine, and 40 mg ketamine. Physiologic and behavioral monitoring was performed during the second expedition on 15 female muskoxen. The observed heart rates were 35-58 beats/min and respiratory rates were 25-30 breaths/min. Mean arterial pressures measured using oscillometry ranged between 117-142 mmHg. Pulse oximeter readings ranged from 91-98% with oxygen supplementation, nasal end-tidal carbon dioxide values were 24-42 mmHg, and rectal temperature ranged from 38.9-39.6°C. Induction time was 6-8 min, recovery time 2-6 min after reversal, and duration of anesthesia was 50-100 min. This anesthetic regime thus provided reliable immobilization with minimal pathophysiologic alterations.

REPEATED ANESTHESIA IN A BLACK RHINOCEROS ( DICEROS BICORNIS) TO MANAGE UPPER RESPIRATORY OBSTRUCTION.

This report describes weekly repeated anesthesia in a 7-yr-old, 1,030 kg, female Eastern black rhinoceros ( Diceros bicornis michaeli), that was immobilized six times using a combination of 2 mg etorphine (0.002 mg/kg), 5 mg medetomidine (0.005 mg/kg), 25 mg midazolam (0.024 mg/kg), and 300 mg ketamine (0.29 mg/kg) delivered intramuscularly (IM) via remote dart to facilitate long-term medical care of a bilateral, obstructive Actinomyces sp. rhinitis. The drug combination described in this study resulted in reliable, rapid recumbency of the animal within 2-8 min after initial administration via dart and produced deep anesthesia for 34-78 min without supplemental anesthetic administration. Antagonist drugs (100 mg naltrexone [0.1 mg/kg] and 25 mg atipamezole [0.024 mg/kg] IM) produced reliable and uneventful recoveries in all the procedures. During each anesthetic procedure, the animal was intubated and provided intermittent positive pressure ventilation with a megavertebrate demand ventilator. Tachycardia and hypoxia noted after induction resolved after positive pressure ventilation with oxygen. This report provides useful information on a novel anesthetic protocol used repeatedly for intensive medical management in a black rhinoceros.

EFFECTS OF A SUPPLEMENTAL ETORPHINE DOSE ON PULMONARY ARTERY PRESSURE AND CARDIAC OUTPUT IN IMMOBILIZED, BOMA-HABITUATED WHITE RHINOCEROS ( CERATOTHERIUM SIMUM): A PRELIMINARY STUDY.

The effects of etorphine on the pulmonary vascular system of white rhinoceros ( Ceratotherium simum) have not been described and could play a role in the severe hypoxemia that develops after immobilization with etorphine-based drug combinations. Characterization of these effects requires measurement of pulmonary vascular pressures and cardiac output (CO). To refine a technique for pulmonary arterial catheterization, five boma-habituated white rhinoceros (three females and two males weighing 1,012-1,572 kg) were immobilized by remote injection with etorphine plus azaperone followed by butorphanol. This afforded the opportunity to perform a pilot study and acquire preliminary measurements of pulmonary arterial pressure (PAP) and CO before and after supplemental etorphine given intravenously. Ultrasonographic guidance was used to insert a sheath introducer into a linguofacial branch of a jugular vein. A 160-cm-long pulmonary artery catheter with a balloon and thermistor was then passed through the introducer and positioned with its tip in the pulmonary artery. It was not long enough to permit wedging for measurement of pulmonary artery occlusion pressure. Mean PAP was 35 mm Hg (minimum, maximum 32, 47 mm Hg) and increased ( P = 0.031) by 83% (28, 106%) after supplemental etorphine. Thermodilution CO was 120 L/min (92, 145 L/min) and increased 27% (3, 43%) ( P = 0.031). Heart rate was 100 (88, 112) beats/min and increased 20% (4, 45%) ( P = 0.031), whereas arterial partial pressure of oxygen was 35 mm Hg (30, 94 mm Hg) and decreased 47% (20, 72%) ( P = 0.031). The cardiovascular observations could result from etorphine-induced generalized sympathetic outflow, as has been reported in horses. Further studies of etorphine in isolation are needed to test this suggestion and to discern how the changes in pulmonary vascular pressures and blood flow might relate to hypoxemia in etorphine-immobilized white rhinoceros.

The hairy lizard: heterothermia affects anaesthetic requirements in the Arabian oryx (Oryx leucoryx).

OBJECTIVE: To study the effect of heterothermia on anaesthetic drug requirements in semi-free ranging Arabian oryx and to assess the temperature quotient (Q10) of oxygen consumption. STUDY DESIGN: Prospective observational study and controlled metabolic experiment. ANIMALS: Sixty-eight anaesthetic events in 59 Arabian oryx from Mahazat As-Sayd protected area, Saudi Arabia METHODS: Anaesthesia was induced by remote injection of 25 mg ketamine, 10 mg midazolam and 0.5 mg medetomidine with a variable amount of etorphine based on a target dosage of 20 µg kg-1 and subjective assessment of body mass. Animals not recumbent within 15 minutes or insufficiently anaesthetized were physically restrained and administered supplementary etorphine intravenously depending on the anaesthetic depth. Body temperature (Tb) was measured rectally immediately upon handling of each animal. From six anaesthetized oryx, expiratory gasses for oxygen analysis and metabolic rate calculation were collected at two Tbs; before and after submersion in ice water for approximately 30 minutes. RESULTS: Forty-two animals (62%) became recumbent with the initial dose, with a mean induction time (± standard deviation) of 9 ± 2 minutes. The remaining animals could be handled but needed 0.3 ± 0.1 mg etorphine intravenously to reach the desired level of anaesthesia. There was a significant positive correlation between Tb and effective etorphine dosage (R2 = 0.48, p < 0.0001). Average Tb of the six animals in which metabolic rate was measured decreased from 40.0 ± 0.5°C immediately after induction to 35.5 ± 0.5°C after cooling. This reduction was associated with a reduction in oxygen uptake from 3.11 ± 0.33 to 2.22 ± 0.29 mL O2 minute-1 kg-1, reflected in Q10 of 2.17 ± 0.14. CONCLUSIONS AND CLINICAL RELEVANCE: Tb significantly affects anaesthetic requirements in Arabian oryx and should be considered when selecting dosages for anaesthetic induction for species showing diurnal heterothermy.

SHORT DURATION IMMOBILIZATION OF ATLANTIC WALRUS ( ODOBENUS ROSMARUS ROSMARUS) WITH ETORPHINE, AND REVERSAL WITH NALTREXONE.

Forty adult, male Atlantic walruses ( Odobenus rosmarus rosmarus) were successfully immobilized for the attachment of global positioning system loggers on their tusks and collection of various biological samples. A standard dose of 7.8 mg etorphine was used for each animal, regardless of body size. All animals were reversed with an iv or im injection of 250 mg naltrexone, immediately after tag attachment. Twenty-seven of the animals were intubated and ventilated with 100% oxygen during the recovery period. The induction time was, on average, 4 min 51 sec ± 1 min 46 sec. Several animals had venous pH, and Pco2 levels that indicated severe acidosis and hypercarbia. All animals recovered within an average of 5 min 16 sec ± 2 min 47 sec after reversal. The total time from darting to recovery was 15 min 23 sec ± 3 min 33 sec. The use of naltrexone is recommended for reversal of etorphine immobilization in adult, male walruses, and the use of positive-pressure ventilation with oxygen is highly encouraged.

A RETROSPECTIVE COMPARISON OF CHEMICAL IMMOBILIZATION WITH THIAFENTANIL, THIAFENTANIL-AZAPERONE, OR ETORPHINE-ACEPROMAZINE IN CAPTIVE PERSIAN FALLOW DEER (DAMA DAMA MESOPOTAMICA).

Records of 56 Persian fallow deer (Dama dama mesopotamica) immobilized for translocation were reviewed. Twenty-three were administered 0.05 ± 0.01 (mean ± SD) mg/kg thiafentanil (THIA), 20 were administered 0.045 ± 0.008 mg/kg thiafentanil combined with 0.19 ± 0.03 mg/kg azaperone (THIA-AZP), and 13 were administered 0.032 ± 0.04 mg/kg etorphine-acepromazine (ETOR-ACP) by intramuscular remote injection. Parameters recorded and compared between groups included induction and recovery times, heart rate, respiratory rate, rectal temperature, oxygen saturation, blood pressure, reflexes, quality of immobilization, and blood concentrations of lactate and glucose. Naltrexone (THIA groups) or diprenorphine (ETOR-ACP) were administered for reversal. Mean induction time was significantly shorter in the THIA group versus the ETOR-ACP group (2.0 ± 1.3 and 4.8 ± 2.8 min, respectively), but not significantly shorter than the THIA-AZP group (2.8 ± 3.1 min). Respiratory rate was significantly higher in the THIA group in comparison to the two other groups. None of the protocols provided excellent immobilization quality, which was significantly poorer in the THIA group. Following antagonist administration, all deer from the THIA and ETOR-ACP groups recovered quickly, while there were five perianesthetic morbidity and mortality cases in the THIA-AZP group. Mean recovery time was significantly shorter in the THIA group versus the THIA-AZP and ETOR-ACP groups (0.5 ± 0.3, 1.1 ± 0.8, and 2.3 ± 1.1 min, respectively). In conclusion, the use of THIA provided faster induction and recovery, with less respiratory depression, but poorer immobilization. The THIA-AZP combination should be used with caution in Persian fallow deer until further investigation.

Activation of delta-opioid receptor contributes to the antinociceptive effect of oxycodone in mice.

Oxycodone has been used clinically for over 90 years. While it is known that it exhibits low affinity for the multiple opioid receptors, whether its pharmacological activities are due to oxycodone activation of the opioid receptor type or due to its active metabolite (oxymorphone) that exhibits high affinity for the mu-opioid receptors remains unresolved. Ross and Smith (1997) reported the antinociceptive effects of oxycodone (171nmol, i.c.v.) are induced by putative kappa-opioid receptors in SD rat while others have reported oxycodone activities are due to activation of mu- and/or delta-opioid receptors. In this study, using male mu-opioid receptor knock-out (MOR-KO) mice, we examined whether delta-opioid receptor was involved in oxycodone antinociception. Systemic subcutaneous (s.c.) administration of oxycodone (above 40mg/kg) could induce a small but significant antinociceptive effect in MOR-KO mice by the tail flick test. Delta-opioid receptor antagonist (naltrindole, 10mg/kg or 20mg/kg, i.p.) could block this effect. When oxycodone was injected directly into the brain of MOR-KO mice by intracerebroventricular (i.c.v.) route, oxycodone at doses of 50nmol or higher could induce similar level of antinociceptive responses to those observed in wild type mice at the same doses by i.c.v. Delta-opioid receptor antagonists (naltrindole at 10nmol or ICI 154,129 at 20µg) completely blocked the supraspinal antinociceptive effect of oxycodone in MOR-KO mice. Such oxycodone antinociceptive responses were probably not due to its active metabolites oxymorphone because (a) the relative low level of oxymorphone was found in the brain after systemically or centrally oxycodone injection using LC/MS/MS analysis; (b) oxymorphone at a dose that mimics the level detected in the mice brain did not show any significant antinocieption effect; (c) oxycodone exhibits equal potency as oxymorphone albeit being a partial agonist in regulating [Ca(2+)]I transients in a clonal cell line expressing high level of mu-opioid receptor. These data suggest that oxycodone by itself can activate both the mu- and delta-opioid receptors and that delta-opioid receptors may contribute to the central antinociceptive effect of oxycodone in mice.

EVALUATION OF ETORPHINE AND MIDAZOLAM ANESTHESIA, AND THE EFFECT OF INTRAVENOUS BUTORPHANOL ON CARDIOPULMONARY PARAMETERS IN GAME-RANCHED WHITE RHINOCEROSES (CERATOTHERIUM SIMUM).

Nineteen white rhinoceroses ( Ceratotherium simum ) were anesthetized with 4 mg of etorphine hydrochloride; 35-40 mg of midazolam; and 7,500 international units of hyaluronidase for dehorning purposes at a game ranch in South Africa, to investigate this anesthetic combination. Median time to recumbency was 548 sec (range 361-787 sec). Good muscle relaxation and no muscle rigidity or tremors were observed in 18 animals, and only 1 individual showed slight tremors. In addition, all animals received butorphanol i.v. 5 min after recumbency at the ratio of 10 mg of butorphanol per 1 mg of etorphine. Blood gas and selected physiologic parameters were measured in the recumbent animal, immediately before and 10 min after the administration of butorphanol. Statistically significant improvements were observed in blood gas physiologic and cardiopulmonary parameters 10 min after the administration of butorphanol, with a reduction in arterial partial pressure of carbon dioxide, systolic blood pressure, and heart rate and an increase in pH, arterial partial pressure of oxygen, oxygen saturation, and respiratory rate (all P < 0.005). After i.v. naltrexone reversal, recovery was uneventful, and median time to walking or running was 110 sec (range 71-247 sec). The results indicate etorphine and midazolam combination is an effective alternative anesthetic protocol and produces good muscle relaxation. Furthermore, i.v. butorphanol was associated with improved blood gas values and cardiopulmonary function for at least 10 min postinjection.

Accelerated induction of etorphine immobilization in blue wildebeest (Connochaetes taurinus) through the addition of hyaluronidase.

OBJECTIVE: To study the effects of the addition of hyaluronidase (HA) to an etorphine/azaperone drug combination on induction times of immobilization. STUDY DESIGN: Experimental part-randomized 'blinded' cross-over study. ANIMALS: Eight wild managed blue wildebeest (Connochaetes taurinus). METHODS: Animals were immobilized, on separate occasions separated by two weeks, with one of four treatments. Treatments were; 'Control drugs (CD), etorphine 0.01 mg kg(-1)  + azaperone at 0.1 mg kg(-1) ; treatment 1 CD + 5000IU HA; treatment 2 CD + 7500 IU HA; and treatment 3 etorphine 0.007 mg kg(-1)  + azaperone at 0.07 mg kg(-1)  + 7500 IU HA. Times to first effect and to immobilization (from darting to possible to approach and blindfold) were measured. anova was used to compare treatments. Results are given in means ± SD (range). RESULTS: For control, and treatments 1-3 respectively, times (in minutes) to first effect were 1.58 ± 0.42 (1.02-2.10), 1.64 ± 0.42 (0.95-2.17), 1.12 ± 0.24 (0.80-1.48) and 1.60 ± 0.21 (1.13-1.88) and to immobilization were 5.38 ± 1.53 (3.82-8.07), 3.80 ± 1.14 (2.02-5.50), 3.51 ± 1.08 (2.28-5.52) and 4.46 ± 0.67 (3.30-5.40). Compared to control, time to first effect for treatment 2 was significantly shorter. Time to immobilization was significantly quicker in all three treatments containing HA than that for control. CONCLUSION AND CLINICAL RELEVANCE: Hyaluronidase can reduce the time to immobilization when used in the immobilizing dart, and might be usefully incorporated into etorphine combinations for darting wildlife.

ETORPHINE-KETAMINE-MEDETOMIDINE TOTAL INTRAVENOUS ANESTHESIA IN WILD IMPALA (AEPYCEROS MELAMPUS) OF 120-MINUTE DURATION.

There is a growing necessity to perform long-term anesthesia in wildlife, especially antelope. The costs and logistics of transporting wildlife to veterinary practices make surgical intervention a high-stakes operation. Thus there is a need for a field-ready total intravenous anesthesia (TIVA) infusion to maintain anesthesia in antelope. This study explored the feasibility of an etorphine-ketamine-medetomidine TIVA for field anesthesia. Ten wild-caught, adult impala ( Aepyceros melampus ) were enrolled in the study. Impala were immobilized with a standardized combination of etorphine (2 mg) and medetomidine (2.2 mg), which equated to a median (interquartile range [IQR]) etorphine and medetomidine dose of 50.1 (46.2-50.3) and 55.1 (50.8-55.4) µg/kg, respectively. Recumbency was attained in a median (IQR) time of 13.9 (12.0-16.5) min. Respiratory gas tensions, spirometry, and arterial blood gas were analyzed over a 120-min infusion. Once instrumented, the TIVA was infused as follows: etorphine at a variable rate initiated at 40 µg/kg per hour (adjusted according to intermittent deep-pain testing); ketamine and medetomidine at a fixed rate of 1.5 mg/kg per hour and 5 µg/kg per hour, respectively. The etorphine had an erratic titration to clinical effect in four impala. Arterial blood pressure and respiratory and heart rates were all within normal physiological ranges. However, arterial blood gas analysis revealed severe hypoxemia, hypercapnia, and acidosis. Oxygenation and ventilation indices were calculated and highlighted possible co-etiologies to the suspected etorphine-induced respiratory depression as the cause of the blood gas derangements. Impala recovered in the boma post atipamezole (13 mg) and naltrexone (42 mg) antagonism of medetomidine and etorphine, respectively. The etorphine-ketamine-medetomidine TIVA protocol for impala may be sufficient for field procedures of up to 120-min duration. However, hypoxemia and hypercapnia are of paramount concern and thus oxygen supplementation should be considered mandatory. Other TIVA combinations may be superior and warrant further investigation.

COMPARISON OF ETORPHINE-ACEPROMAZINE AND MEDETOMIDINE-KETAMINE ANESTHESIA IN CAPTIVE IMPALA (AEPYCEROS MELAMPUS).

Impala (Aepyceros melampus) are a notoriously difficult species to manage in captivity, and anesthesia is associated with a high risk of complications including mortality. The aim of this study was to compare an opioid-based protocol with an α-2 agonist-based protocol. Nine female impala were studied in a random cross-over design. Subjects received either an etorphine-acepromazine (EA) protocol: 15 µg/kg etorphine and 0.15 mg/kg acepromazine, or a medetomidine-ketamine (MK) protocol: 109 µg/kg medetomidine and 4.4 mg/kg ketamine on day 1. Anaesthesia was repeated 3 days later with the alternative protocol. Subjective assessments of the quality of induction, muscle relaxation, and recovery were made by a blinded observer. Objective monitoring included blood pressure, end-tidal CO2, regional tissue oxygenation, and blood gas analysis. EA provided a significantly quicker (mean EA, 7.17 mins; MK, 17.6 mins) and more-reliable (score range EA, 3-5; MK, 1-5) induction. Respiratory rates were lower for EA with higher end-tidal CO2, but no apnoea was observed. As expected, blood pressures with EA were lower, with higher heart rates; however, arterial oxygenation and tissue oxygenation were equal or higher than with the MK protocol. In conclusion, at these doses, EA provided superior induction and equivalent muscle relaxation and recovery with apparent improved oxygen tissue delivery when compared to MK.

Effects of Butorphanol on Respiration in White Rhinoceros (Ceratotherium simum) Immobilized with Etorphine-Azaperone.

This article reports on respiratory function in white rhinoceros (Ceratotherium simum) immobilized with etorphine-azaperone and the changes induced by butorphanol administration as part of a multifaceted crossover study that also investigated the effects of etorphine or etorphine-butorphanol treatments. Six male white rhinoceros underwent two immobilizations by using 1) etorphine-azaperone and 2) etorphine-azaperone-butorphanol. Starting 10 min after recumbency, arterial blood gases, limb muscle tremors, expired minute ventilation, and respiratory rate were evaluated at 5-min intervals for 25 min. Alveolar to arterial oxygen gradient, expected respiratory minute volume, oxygen consumption, and carbon dioxide production were calculated. Etorphine-azaperone administration resulted in hypoxemia and hypercapnia, with increases in alveolar to arterial oxygen gradient, oxygen consumption, and carbon dioxide production, and a decrease in expired minute ventilation. Muscle tremors were also observed. Intravenous butorphanol administration in etorphine-azaperone-immobilized white rhinoceros resulted in less hypoxemia and hypercapnia; a decrease in oxygen consumption, carbon dioxide production, and expired minute ventilation; and no change in the alveolar to arterial oxygen gradient and rate of breathing. We show that the immobilization of white rhinoceros with etorphine-azaperone results in hypoxemia and hypercapnia and that the subsequent intravenous administration of butorphanol improves both arterial blood oxygen and carbon dioxide partial pressures.