Reversible immobilization of free-ranging African lions (Panthera leo) with medetomidine-tiletamine-zolazepam and atipamezole

A combination of medetomidine-tiletamine-zolazepam was used to conduct six immobilizations of free-ranging lions (Panthera leo) in Waza National Park, Cameroon, during 1999 and 2000. Drugs were administered by dart injection at 0.07+/-0.01 (mean+/-SD) mg/kg of medetomidine and 1.8+/-0.5 mg/kg of tiletamine-zolazepam. Chemical immobilization was characterized by smooth inductions (14.1+/-6 min), satisfactory analgesia, and muscle relaxation. One animal was treated for bradypnea. No major alterations of physiologic parameters (heart and respiratory rates, rectal temperature) were seen during immobilization in the other lions. Relative arterial oxygen saturation was measured in two animals and revealed mild hypoxemia. The animals received atipamezole at 0.3+/-0.1 mg/kg intramuscularly for reversal of anesthesia. Recoveries were uneventful. All animals were radiocollared, and no mortalities occurred during an 18-mo follow-up period. Use of medetomidine-tiletamine-zolazepam for anesthesia and reversal of anesthesia with atipamezole appear to be useful for reversible immobilization of free-ranging lions.     

Anti-stress effect of dalargin in immobilization stress in rats

The function of the isolated perfused rat hearts was studied in four groups of experiments. Group 1--included the hearts of intact animals ("absolute control"), group 2--the hearts of rats subjected to 24 hour immobilization in supine position against the background of triple intramuscular injections of placebo (control), group 3 included the hearts of rats, which during 24 hour immobilization stress were thrice injected a synthetic analogue of endogenous opioids Dalargin in a dose of 3 g/kg, and group 4 included the hearts of animals, which during immobilization were administered Dalargin in a dose of 10 g/kg of body mass. Ulcer index as indicator of stress injury of gastric mucosa was also determined. In the control group of experiments (group 2) 24 hour immobilization stress resulted in complete depression of cardiac performance as compared with group 1, and ulcer index approximated 1. In group 3 the indices of cardiac performance even exceeded those in group 1 (intact animals). As compared with group 2, ulcer index in group 3 decreased by 9 times. In application of Dalargin in a dose of 10 g/kg complete preservation of heart function indices and complete prevention of stress injury of gastric mucosa were also observed. Thus, Dalargin possesses cardioprotective and anti-ulcerogenic effect in immobilization stress in rats. Most probably, this phenomenon can be attributed to its ability to inhibit the activity of sympathoadrenal system, which gets enhanced during stress.     

Individual characteristics of the nociceptive sensitivity of rats during the formation of a negative emotional reaction

Changes in dynamics of nociceptive sensitivity in male rats of Wistar strain were studied during formation of negative emotional reaction. The procedure of blood's taking from the tail after its tip's amputation was used as negative emotional factor. The animals were divided into six groups by the criterion of individual properties of nociceptive sensitivity dynamics. On the whole it was found to decrease. Besides, one more group (the seventh group) was found with profound hypoalgesia caused by action of the negative emotional factor. It was also shown that formation of negative emotional reaction in two groups was not accompanied by changes of nociception. The data suggest a selective influence of negative emotional factor on nociceptive sensitivity.     

Evaluation of xylazine hydrochloride as the sole immobilizing agent in moose and caribou--and its subsequent reversal with idazoxan

Xylazine hydrochloride was used as the sole immobilizing agent in moose and caribou. The animals were free-ranging and immobilization was accomplished from a helicopter using powered darts. Following a period of immobilization during which radiotelemetry collars were fitted, the animals were revived using idazoxan (RX 781094) or its methoxy analogue RX 821002. Xylazine was administered at dose rates of approximately 3.0 mg/kg and 5.0 mg/kg to the moose and caribou, respectively. Moose received 430 +/- 27 mg of xylazine and a mean dose of 10 mg idazoxan (RX 781094). Caribou received 485 +/- 30 mg xylazine and a mean dose of 4 mg idazoxan (RX 821002). This technique gave adequate immobilization with rapid recovery of consciousness in both species.     

Hibernation and disuse osteoporosis.

In an attempt to determine 1) if disuse osteoporosis occurs naturally during hibernation and 2) if it can be induced by limb immobilization in active and hibernating ground squirrels, forty-six thirteenlined ground squirrels were divided into five experimental groups. Group one was the active control group (AC) (ten animals). Group two was the active immobilized (AI) group and was subjected to a unilateral sciatic nerve section (ten animals). Group three was the hibernating control group and was allowed to hibernate undisturbed for 30 days (HC) (ten animals). The fourth group was subjected to a unilateral sciatic nerve section and allowed to hibernate for 30 days; this was the hibernating immobilized group (HI). The fifth group of six animals received the identical treatment as the (HI) group but was allowed to hibernate for 150 days.The following measurements were taken: Gastrocnemius-soleus mass, body mass, tibia-fibula weight, total tibia-fibula calcium and phosphorus. In addition, the femur was observed histologically for the enlarged lacunae typical of osteoporotic bone. Statistical analysis was accomplished with the Wilcoxan sign rank test.The data were interpreted to mean: 1) Disuse osteoporosis can be induced in active ground squirrels, 2) disue osteoporosis is not evident nor can it be induced by limb immobilization during 30 days of hibernation and 3) limb immobilization did not result in significant disuse osteoporosis following 150 days of hibernation. Only a few enlarged lacunae were evident after 150 days of hibernation.     

Modulating effect of opioid peptides on hemopoiesis in stress

The influence of leu-enkephalin and dalargin on the blood system was studied during immobilization stress in mice. The early transmitted reactions of the peripheral blood were shown to decrease upon single drug infusions after immobilization. At later terms the activation of bone marrow hematopoiesis was not registered in mice receiving opioid peptides in contrast to the control animals. It correlates with drug-induced decrease in the mitotic activity of bone-marrow cells. Suppressive effect of opioids on hematopoiesis during stress was connected with their decreasing effect on corticosteroid level in the animal plasma. The latter can suggest indirect influence of enkephalins on bone marrow hematopoiesis in immobilization stress.     

Repeated immobilization stress increases total cytosolic glucocorticoid receptor in rat liver

Glucocorticoids are well-known mediators of stress-related endocrine, autonomic, and behavioral responses in mammals and human beings. However, our understanding of the mechanisms of glucocorticoid action in response to stress remains elusive. Therefore, in the present study, an effort has been made to systematically examine glucocorticoid action during acute (2 h) and repeated (2 h daily for 7, 15, and 30 days) immobilization stress in male Sprague-Dawley rats. Prolonged 30-day stress resulted in reduced total body weight gain. There was a dramatic 3- to 4-fold increase in plasma corticosterone levels after single acute stress paradigm, which remained augmented 2- to 3-fold higher than basal control levels during the repeated 30-day immobilization conditions. There was good relationship between increased plasma corticosterone levels and elevation of tyrosine aminotransferase activity in the liver during 30 days of stress. Because repeated immobilization stress animals showed increased levels of both plasma corticosterone and tyrosine aminotransferase activity, the regulation of cytosolic glucocorticoid receptor (GR) in rat liver, a major target tissue for glucocorticoid, was carried out during repeated stress by using GR binding assay, exchange assay, and Western blotting techniques. Exposure of animals to acute and repeated stress resulted in decreased free cytosolic GR. Interestingly, the bound cytosolic GR increased remarkably in liver during prolonged stress of 7-30 days. Overall, results obtained by using both binding assays and Western blotting for the first time showed that repeated stress animals had higher levels of total hepatic cytosolic GR as compared to control animals. These novel results suggest that repeated stress influences the hypothalamic-pituitary-adrenal axis in rats by elevating both the level of plasma corticosterone and total hepatic cytosolic GR.     

A computational model for the adaptation of muscle and tendon length to average muscle length and minimum tendon strain

This paper hypothesizes that average muscle length and minimum tendon strain govern muscle and tendon length adaptation in all situations. A model has been implemented to test this hypothesis, and simulations have been performed for normal development, bone lengthening, immobilization, and retinacular release experiments in young and adult animals. The simulation results predict that both muscle and tendon lengthen during normal development, with the rate of tendon growth slowing faster than the rate of muscle growth. The results also predict that muscle length increases during bone lengthening in both young and adult animals, while tendon length increases only in young animals. For immobilization in adult animals, the results predict that muscle length increases when the muscle is immobilized in a lengthened position and decreases when the muscle is immobilized in a shortened position with no change in tendon length. For immobilization in young animals, the results predict reduced muscle growth and increased tendon growth regardless of immobilization position. Finally, the simulations predict that retinacular release which increases excursion of the musculotendinous unit leads to increased muscle length with decreased tendon length in young animals and decreased muscle length with no change in tendon length in adult animals. These simulation results are consistent with experimental findings reported in the literature by other investigators. This suggests that average muscle length and minimum tendon strain may represent general principles that govern muscle and tendon length adaptation.     

Efficiency and safety of xylazine and tiletamine/zolazepam to immobilize captured wild boars (<Emphasis Type="Italic">Sus scrofa</Emphasis> L. 1758): analysis of field results

During a 2-year study on wild boar ecology in a semiagricultural area of Northern Apennines (central Italy), 47 animals were caught with traps and subsequently immobilized with chemical restrain based on a 1:1 mixture of xylazine and tiletamine/zolazepam (1 ml contains: 50 mg of xylazine, 25 mg of tiletamine, and 25 mg of zolazepam). Effective chemical immobilization was observed in 55% of the caught wild boars, and a high capture mortality rate occurred (in 10.6% of wild boars). A posteriori analysis, by means of a logistic regression model, was performed to assess potential factors associated with wild boar chemical immobilization failure. Underdosage and increase in the number of animals caught together significantly increase the wild boar chemical restrain failure, while the capture mortality appears to be positively related to the sole trap size alone. The stress developed during the capture plays a relevant role influencing the outcomes of the anesthesia. Thus, the use of alternative handling or catching procedures with the aim to reduce stress in captured animals could maximize the safety and the efficiency of the wild boar chemical immobilization.     

Field immobilization of feral 'Judas' donkeys (Equus asinus) by remote injection of medetomidine and ketamine and antagonism with atipamezole

The Judas technique is a method used for landscape control of feral donkeys (Equus asinus) in northern Australia. Central to the success of any Judas program is the safe, efficient, and humane attachment of the telemetry device. For feral donkeys, this involves the use of field immobilization. We examine the replacement of the current chemical capture agent, succinylcholine, with contemporary immobilization agents to achieve positive animal welfare outcomes. A combination of medetomidine and ketamine delivered by remote injection from a helicopter was used to capture 14 free-ranging feral donkeys for the fitting of telemetry collars in Western Australia in November 2010. Dose rates of 0.14 mg/kg medetomidine and 4.1 mg/kg ketamine were appropriate to immobilize animals in 9 min (± SD = 3). Mean recovery time (total time in recumbency) was 21 min (± 14). All animals recovered uneventfully after being administered atipamezole, a specific antagonist of medetomidine, intramuscularly at 0.35 mg/kg. Physiologic parameters were recorded during recumbency, with environment-related hyperthermia being the only abnormality recognized. No significant complications were encountered, and this drug combination represents an efficient approach to capturing wild donkeys. This new method allows a rapid, safe, cost-effective approach to the immobilization of feral donkeys for use as Judas animals. This drug combination will replace the relatively inhumane succinylcholine for the field immobilization of feral donkeys.     

Effect of exogenous heparin on secretory status of rat mast cells under immobilization stress]

The significant increase of heparin release from mast cells was observed in rats under stress conditions induced by 60 min immobilization. The index of its saturation with heparin became 4 times lower. The highest secretory activity of mast cells was observed during the first 30 min of immobilization. It was shown that at that time the heparin release from mast cells occurred by granulolysis (merocrine type of secretion). In the rats received heparin (15 or 150 u/200 g) during the first 15 min of immobilization the mast cells released heparin with the same intensity as in a 4 control animals. But then in rats with high heparin blood concentration the heparin release from mast cells ceased and mast cells began to accumulate heparin from blood. By the 30th min of immobilization the heparin content in the mast cells has become normal.     

Pain Modulation in Waking and Hypnosis in Women: Event-Related Potentials and Sources of Cortical Activity

Using a strict subject selection procedure, we tested in High and Low Hypnotizable subjects (HHs and LHs) whether treatments of hypoalgesia and hyperalgesia, as compared to a relaxation-control, differentially affected subjective pain ratings and somatosensory event-related potentials (SERPs) during painful electric stimulation. Treatments were administered in waking and hypnosis conditions. LHs showed little differentiation in pain and distress ratings between hypoalgesia and hyperalgesia treatments, whereas HHs showed a greater spread in the instructed direction. HHs had larger prefrontal N140 and P200 waves of the SERPs during hypnotic hyperalgesia as compared to relaxation-control treatment. Importantly, HHs showed significant smaller frontocentral N140 and frontotemporal P200 waves during hypnotic hypoalgesia. LHs did not show significant differences for these SERP waves among treatments in both waking and hypnosis conditions. Source localization (sLORETA) method revealed significant activations of the bilateral primary somatosensory (BA3), middle frontal gyrus (BA6) and anterior cingulate cortices (BA24). Activity of these contralateral regions significantly correlated with subjective numerical pain scores for control treatment in waking condition. Moreover, multivariate regression analyses distinguished the contralateral BA3 as the only region reflecting a stable pattern of pain coding changes across all treatments in waking and hypnosis conditions. More direct testing showed that hypnosis reduced the strength of the association of pain modulation and brain activity changes at BA3. sLORETA in HHs revealed, for the N140 wave, that during hypnotic hyperalgesia, there was an increased activity within medial, supramarginal and superior frontal gyri, and cingulated gyrus (BA32), while for the P200 wave, activity was increased in the superior (BA22), middle (BA37), inferior temporal (BA19) gyri and superior parietal lobule (BA7). Hypnotic hypoalgesia in HHs, for N140 wave, showed reduced activity within medial and superior frontal gyri (BA9,8), paraippocampal gyrus (BA34), and postcentral gyrus (BA1), while for the P200, activity was reduced within middle and superior frontal gyri (BA9 and BA10), anterior cingulate (BA33), cuneus (BA19) and sub-lobar insula (BA13). These findings demonstrate that hypnotic suggestions can exert a top-down modulatory effect on attention/preconscious brain processes involved in pain perception.     

Chemical sensitivity of neurons to immobilization emotional stress in the rat

Chemical sensitivity of the neurones of the medial thalamus (MT) and ventromedial hypothalamus (VMH) to noradrenaline (NA) was studied microionophoretically in conditions of 30-hour immobilization of rats. Chemical sensitivity of single neurons of MT as well as VMH during immobilization is characterized by a certain instability, showed in changes of their reactions at repeated NA applications. These changes were observed significantly more frequently in MT neurones especially at the initial stage of immobilization. At the final stage as well as in unrestrained control animals, changes of the sensitivity were seen significantly more seldom. At the initial stage of rats' immobilization a significant increase of the number of MT and VMH cells responding to NA application by the excitative type of reactions was also observed.     

Time course of vasopressin and oxytocin secretion after stress in adrenalectomized rats.

To characterize the participation of vasopressin (AVP) and oxytocin (OT) in hypothalamus-pituitary-adrenal regulation after adrenalectomy (ADX), we evaluated corticosterone, ACTH, AVP and OT plasma concentrations and AVP and OT content of the paraventricular nucleus (PVN) at different periods (3 h, 1, 3, 7 and 14 days) in sham or ADX rats under basal conditions and after immobilization stress. ADX animals showed undetectable corticosterone levels, while sham animals showed a marked increase in corticosterone and ACTH 3 h after surgery, then lowering to basal control levels. ADX rats showed high basal ACTH levels with a triphasic response without changes after immobilization. After three hours, the ADX group showed higher OT levels than the sham group. OT was increased after immobilization stress in sham and ADX groups. AVP plasma levels did not change throughout the basal or stress studies in either group. There was a decrease in hypothalamic AVP content 1 and 3 days after ADX under basal and stress conditions. Plasma osmolality showed a significant decrease in the ADX group at 3, 7, and 14 days. In conclusion, there are different pituitary-adrenal axis set points after removal of the glucocorticoid negative feedback. The role of vasopressinergic and oxytocinergic neurons in the ACTH secretion after ADX or immobilization stress appears to differ. Magnocellular AVP is unlikely to contribute to ACTH secretion in response to ADX or immobilization stress. On the other hand, OT is elicited by immobilization stress and might contribute to the ACTH secretion during short-term ADX.     

Influence of surgical pain stress on the blood-brain barrier permeability in rats

Effect of surgical pain stress on the blood-brain barrier permeability was investigated in rats. The animals were divided into four groups: Group 1: control, Group 2: immobilization stress, Group 3: acute hypertension, Group 4: immobilization stress + surgical pain stress.Bilateral hid paw surgical wounds for cannulations were applied in animals' inguinal regions under diethyl-ether anesthesia, then the animals were awaken from anesthesia to produce surgical pain stress. Evans-blue was used as a blood-brain barrier tracer. There is no significantly blood-brain barrier breakdown after short-time immobilization stress, but after adrenalin hypertension blood-brain barrier permeability was increased especially on frontal and occipital cortices in 50% of the animals. Surgical pain stress increased blood-brain barrier permeabiliy in comparison to acute adrenalin-induced hypertension (p < 0.01). In surgical pain stress-induced animals distinct Evans-blue leakage was observed in the occipital, frontal and parieto-temporal cortices.     

Cytoplasmic superoxide dismutase activity is a sensitive indicator of the antioxidant status of the rat liver and brain

Several parameters of the cytoplasmic enzymatic antioxidant system of the liver and brain of the rat have been investigated under conditions of immobilization stress and of an antioxidant preparation in the diet of animals. These included superoxide dismutase (SOD) and glutathione reductase (GR) activities and nonspecific NADPH oxidation. Only changes in the activity of SOD both in the liver and brain were revealed. In the liver of animals that receive no preparation, a decrease in the activity of SOD after 30-min immobilization and its restoration after a 360-min immobilization were observed. In the brain, the activity of SOD decreased only in preconditioned animals after 30 and 360 min of exposure to stress. In addition, the activity of SOD in the brain of preconditioned animals, both stressed and unstressed, was lower than in the corresponding groups of control animals. It is probable that, under the conditions of immobilization stress, the level of reactive oxygen species (ROS) and as a consequence the activity of SOD decrease. The intake of an antioxidant preparation under these conditions seems to be not correct.     

Mitotic activity of the lymphocytes of the thymus cortex in hypokinesia during the period of readaptation

The changes in the weight and mitotic index were studied in the cortex of the thymus of Wistar rats during 10-day hypokinesia and 10-day readaptation (restoration). 24 hours after immobilization of the animals the mitotic index was 2 times as lower. No complete readaptation was attained during 10-day hypokinesia. No readaptation was attained during 10-day hypokinesia. In readaptation the stage of secondary stress was found (the mitotic index was 3.5 times as reduced), the stage of genuine restoration being revealed after 10 days.     

Role of the pituitary-adrenal system in mechanism regulating vascular permeability during stress

Disturbances of the microvascular permeability were studied by the "vascular labelling" technique during the immobilization stress of hypophysectomized and adrenalectomized rats. Animals with sham operations served as controls. As revealed, hypophysectomy and adrenalectomy caused disturbances of vascular permeability in the mesentery. Vascular permeability disturbances in the hypophysectomized and adrenalectomized rats under conditions of immobilization were more expressed than in the sham-operated animals. Removal of the pituitary and adrenal glands produced mast cell degranulation at the earlier immobilization period.     

Effects of immobilization stress on tuberoinfundibular dopaminergic (TIDA) neuronal activity and prolactin levels in lactating and non-lactating female rats.

The effects of immobilization stress on the prolactin secretory response and on the activity of the tuberoinfundibular dopaminergic (TIDA) neurons were determined in intact, virgin female rats on the morning of diestrus or proestrus and in post-partum, lactating female rats. The virgin females exhibited a significant increase in circulating levels of prolactin which was evident by 1 minute and persisted during the immobilization (5 minutes). In contrast, the prolactin secretory response in lactating females was significantly attenuated compared to non-lactating animals. The activity of the TIDA neurons was not altered by the 5 minutes of stress. Even after 30 minutes of immobilization, TIDA neuronal activity was not affected in either the lactating or cycling females. These data suggest that the cycling female rat is capable of a prolactin secretory response to the stressor without inhibition of TIDA neuronal activity. It seems likely that prolactin releasing factors mediate this response. In contrast, stress did not produce a similar prolactin increase during lactation. It seems likely that, during lactation, the pituitary is not sensitive to releasing factors unless the TIDA neurons are inhibited. There appear to be differences in the sensitivity of the pituitary depending on the physiological state of the model employed.     

Relative efficiency of succinylcholine, xylazine, and carfentanil/xylazine mixtures to immobilize free-ranging moose

We compared the efficiency of succinylcholine chloride, xylazine hydrochloride and carfentanil/xylazine mixtures in immobilizing 364 free-ranging moose (Alces alces) between 1987 and 1997 in Québec (Canada). With succinylcholine chloride (0.070, 0.062, 0.051 mg/kg of estimated body weight for calves, juveniles and adults), 63% of the 252 immobilization attempts led to complete immobilization and marking, whereas 7% of the darted animals died of respiratory paralysis during handling. The moose took an average of 13 min to lay down after darting (down time). Injection of xylazine (3.67-4.22 mg/kg) permitted sedation (the animal laid down but got up again when approached) or complete immobilization in 78% of the 40 darted adult moose, the mean down time being 8.7 min. No mortality was noted with this drug but 58% of the marked animals were only sedated. The use of RX821002A (0.058 mg/kg) as an antagonist, permitted a mean recovery time of 2.8 min after intravenous injection. With the carfentanil/xylazine mixtures (0.0071 and 0.181 mg/kg), 96% of the immobilization trials (n = 72) led to complete (88%) or partial (8%) immobilization, but 6% of the moose died several days after capture. The mean down time was 6.6 min, and injection of naltrexone (0.709 mg/kg) antagonized the effect of the immobilizing agent within 3.7 min. The respiratory rate was higher (P < 0.05) among moose immobilized with xylazine (35/min) than among those immobilized with carfentanil/xylazine mixtures (19/min) but this variation could be related to a longer pursuit time (z = 3.60; P < 0.01) and higher stress levels during handling. Rectal temperature also was higher with xylazine but the difference was small (39.7 vs. 39.3, P = 0.03) and did not differ significantly between the sexes (P > 0.05). Considering loss of materials and helicopter flight time due to non-successful marking trials, carfentanil/xylazine mixtures were the least expensive ($333 Cdn/animal).