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UntitledJournal of Wildlife Diseases, 49(2), 2013, pp. 338–346 REVERSIBLE IMMOBILIZATION OF FREE-RANGING SNOWLEOPARDS (PANTHERA UNCIA) WITH A COMBINATION OFMEDETOMIDINE AND TILETAMINE-ZOLAZEPAM ¨ rjan Johansson,1,2,3,8 Jonas Malmsten,4,5 Charudutt Mishra,3,6 Purevjav Lkhagvajav,7 and Tom McCarthy21 Grimso¨ Wildlife Research Station, Department of Ecology, Swedish University of Agricultural Sciences, SE-73091,Riddarhyttan, Sweden2 Panthera, 8 W 40th Street, 18th floor, New York, New York 10018, USA3 Snow Leopard Trust, 4649 Sunnyside Avenue North, Seattle, Washington 98103, USA4 Department of Pathology and Wildlife Diseases, National Veterinary Institute, SE-751 89 Uppsala, Sweden5 Department of Clinical Sciences, Swedish University of Agricultural Sciences, SE-75007 Uppsala, Sweden6 Nature Conservation Foundation, 3076/5, IV Cross Gokulam Park, Mysore, India7 Snow Leopard Conservation Foundation, Sukhbaatar District, 4th Khoroo, 53-9 Ulan Baatar, Mongolia8 Corresponding author (email: firstname.lastname@example.org) Conservation and research of the elusive snow leopard (Panthera uncia) have been hampered by inadequate knowledge about its basic life history. Global positioning system (GPS)collars can provide useful information, but there has been limited information available on safecapture methods, drug doses, and efficacy for effective immobilization of free-ranging snowleopards. We describe a drug protocol using a combination of medetomidine and tiletamine-zolazepam for the chemical immobilization of free-ranging snow leopards. We also describephysiologic responses to immobilization drugs, including rectal temperature, heart rate,respiratory rate, and relative hemoglobin oxygen saturation (SpO2) recorded every 10 min. Our study was carried out in the Tost Mountains adjacent to the Great Gobi Desert, in southernMongolia, between August 2008 and April 2012. Eighteen snow leopards were captured orrecaptured with foot-snares on 42 occasions and anesthetized for marking with GPS collars. Thesnow leopards received on average (6SD) 0.02060.04 mg/kg body mass medetomidine and2.1760.45 mg/kg tiletamine-zolazepam. The duration of ensuing anesthesia was 69613 min,including an induction period of 10 (64) min. Anesthesia was reversed with 4 mg (0.1060.04 mg/kg) atipamezole administered intramuscularly. The mean value for SpO2 for the 37 captures where we could record physiologic values was 9164. The SpO2 increased significantly during anesthesia (+0.0660.02%/min), whereas rectal temperature (average 38.160.7 C/min, change 20.0460.003C/min), heart rate (average 9769 beats/min, change 20.2060.03 beats/min), and respiratoryrate (average 2666 breaths/min, change 20.1160.03 breaths/min) decreased significantly. Adose of 80 mg tiletamine-zolazepam (2 mg/kg body weight) and 0.72 mg medetomidine(0.02 mg/kg body weight) safely immobilized all adult and subadult snow leopards (weight 25–45 kg) in our study. All measured physiologic values remained within clinically acceptablelimits.
Anesthesia, capture, drug dose, Mongolia, physiology, vital rates.
2003). Chemical immobilization must relyon a safe drug, or drug combination, and that inhabit relatively inaccessible terrain sufficient maintenance of physiologic pro- cesses such as respiration and heart rate tracking device, for example, global posi- carnivores, this requires the animals to be in the mountains of Central Asia. Captive important that the immobilization process be safe and involve as little stress to the JOHANSSON ET AL.—REVERSIBLE IMMOBILIZATION OF FREE-RANGING SNOW LEOPARDS (Jalanka, 1989b). The three field studies that involved multiple immobilizations ofwild snow leopards employed ketamine study of snow leopards in Mongolia. A keycomponent of the study was to capture and fit snow leopards with GPS collars (for a detailed description of the study, see McCarthy et al., 2010). The study area was located in the Tost Mongolia. Our primary base camp was locatedat 43u119N, 100u369E. Annual precipitation in strong winds all year, especially in winter and spring. Elevation ranges between 1,900 and conducted August 2008 to April 2012. Snow leopards were caught in modified Aldrich-style Novak, 1980) placed in steep-walled, narrow canyons. The snare design follows Frank et al.
(2003), with the exception that the snares wereanchored with a four-way cross stake ground (tiletamine) and a benzodiazepine agonist stakes,17 mm in diameter, with a nut welded properties (Plumb, 2008). Used alone, it causes to one end. The stakes were hammered in at about 45u angle from the ground and perpen- laryngeal reflexes, and cataleptoid anesthesia.
dicular to one another. This prevented thecaptured animal from escaping with the snare Adverse effects include marked salivation, cable attached to its leg. A spring was also which, in combination with retained laryn- attached between the anchor and the snare to geal reflexes, can cause vomiting and retching struggled. Each snare was equipped with a trap-site VHF transmitter (TBT-500, TelonicsInc., Mesa, Arizona, USA) emitting a contin- and TZ has reportedly been used for safe, uous signal at a unique frequency. A change in efficient, and reversible immobilization of signal pulse rate indicated the trap had been signals every 3 hr from early evening to late morning from a mountain peak using ahandheld receiver (Communications Special- Jacquier et al., 2006), California sea lions Gulland, 2001), captive polar bears (Ursus 4FL, Followit, Lindesberg, Sweden). Starting maritimus; Caulkett et al., 1999), and free- in autumn 2010, we used an automatic trap surveillance system that constantly monitored Caulkett and Cattet, 1997). We report the the traps. The system scans the trap transmit-ters and sounds an alarm within a maximum of 8 min after a snare has been sprung (Johans- son et al., 2011). This greatly reduced our objectives were to evaluate the physiologic response time, allowing us to be at the capture site in an average of 40 min, and a maximum of bilization, and to describe a drug combi- immobilized with a combination of TZ (Tela- effectively immobilize average-sized sub- Fort Dodge Laboratories, Inc., Fort Dodge, JOURNAL OF WILDLIFE DISEASES, VOL. 49, NO. 2, APRIL 2013 Vectronic, Berlin, Germany) programmed to land). The drugs were mixed to a concentra- record the animal’s location every 5 hr and tion of 50 mg zolazepam, 50 mg of tiletamine, uplink the data via Globalstar satellites. Data 5 mL of Domitor to one vial of Telazol (the drug combination henceforth referred to as recorded because we waited out of sight to minimize disturbing the darted snow leopards.
needle (Daninject, Borkøp, Denmark) fired from a CO2-powered rifle (Daninject J.M.
Special) fitted with a red dot sight (Aimpoint and were able to handle the snow leopard.
Once fully sedated, vital rates were measured throughout the anesthesia. Rectal temperature was monitored with a digital thermometer.
leopard and darted it from a distance of 7–15 m, Respiratory rate was measured by counting and the capture site was vacated immediately to the number of chest movements for 30 sec.
avoid stressing the animal. After 6–7 min, the Heart rate and relative hemoglobin oxygen animal was approached to ensure that it was not saturation (SpO2) were measured throughout lying in a position that could obstruct its the anesthesia with a pulse oximeter (Nellcor airways. If unsuccessful anesthesia was con- N65 Oximax handheld pulse oximeter, Nellcor Inc., Boulder, Colorado, USA) by attaching the animal was left for another 5 min. If still sensor (Vetsat, Nellcor Inc.) to the ear.
unanesthetized for any reason (e.g., unsuccess- Capillary refill times were estimated but not ful drug delivery/induction, or dart placement), another full dose was administered, approxi- circulation. To reverse the effect of MED, mately 13–15 min after the first dart.
atipamezole hydrochloride (Antisedan vet 5 mg/mL, Orion Pharma Animal Health) was admin- istered intramuscularly in the quadriceps ortriceps at a dose five times higher than the medetomidine dose. Duration of anesthesia lateral recumbency on an insulated blanket was defined as the time from when the last dart (Fjellduken, Jerven AS, Odda, Norway). If was fired to the time when the snow leopard first raised its head after collaring (head-up time). Time to recovery and time to walking rubber hot-water bottles were placed in the were defined as the time from injection of the groin and axillaries. Eye gel (Viscotears, CIBA reversal drug to the head-up time and the time Vision AG, Hetlingen, Switzerland) was ap- from injection of the reversal drug to the animal walking steadily. In case of a spontaneous blindfolded to protect its eyes from dust and recovery toward the end of the anesthesia, a light. In cases where the capture site was full dose of antidote was nevertheless adminis- deemed unsafe for cat recovery (steep slopes in more than one direction close to the snare),the anesthetized animal was moved to a predetermined handling and awakening site.
The data were analyzed using mixed linear and blood, hair, and ear swabs were collected models in the statistical software R 2.12.1 (R- for serologic, genetic, and parasitologic analy- Development Core Team, 2010) with package ses. Age was estimated based on body weight, lme4. We included snow leopard individuals size, and tooth wear (Stander, 1997). Smaller and capture events as random factors, as some wounds (either capture related or from pre- individuals were captured more than once and capture events) were cleaned with 2% iodine some measurements were taken several times solution, and, if deemed appropriate, a 500-mg during a capture event. Reported mean values of drug dose and physiologic parameters are Norbrook Laboratories Ltd., Carlisle, UK) was for each capture. The effect of time on the administered intramuscularly. Female snow variables was tested with a t-test (df516, leopards were visually inspected for signs of number of individuals: 2), testing whether current or previous lactation. The animals the slope was significantly different from 0.
were equipped with GPS collars (North Star, JOHANSSON ET AL.—REVERSIBLE IMMOBILIZATION OF FREE-RANGING SNOW LEOPARDS Description of individual snow leopards (Panthera uncia) captured and immobilized in Tost Mountains of South Gobi Province, Mongolia, August 2008–April 2012. Snow leopards were weighed using adigital scale with an accuracy of 0.1 kg; hobbles were placed around the front and hind legs, and tworesearchers lifted the animal. Weight is given as a range when the same individual was caught more than once.
Age estimates are based on tooth wear and body size.
a Caught in wolf trap.
b Offspring of F4.
c Offspring of F3, still travelling with his mother at time of capture.
killed M2 3 mo after he was collared. Theother three adult males caught early in the captured 17 snow leopards, nine males and eight females, on 41 occasions. In addition,one male (M6) was anesthetized and released from a steel-jaw wolf trap set by (1.69 mg/kg and 1.36 mg/kg, respectively) this anesthesia were also included in the analysis. Physiologic data were collected for for safety reasons. On those occasions, we M3), resulting in anesthesia durations of 53, 36, and 85 min, respectively. Thereaf- one capture occurred during daytime.
JOURNAL OF WILDLIFE DISEASES, VOL. 49, NO. 2, APRIL 2013 Drug doses and physiologic response of anesthetized snow leopards (Panthera uncia) captured and immobilized in Tost Mountains of South Gobi Province, Mongolia, August 2008–April 2012. Values for t and Pare from t-tests.
a MED 5 medetomidine; TZ 5 tiletamine-zolazepam (doses are mg/kg body weight); SpO2 5 relative oxygen saturation in hemoglobin measured with a pulse oximeter.
b Change in physiologic parameter per minute, derived using mixed linear models; NS 5 not statistically significant.
Induction, duration of anesthesia, and recovery in an attempt to escape. Once darted, and upon the retreat of the darter, the animals rate decreased significantly during anes- generally remained silent, indicating that they remained still during induction.
tion time was not significantly related to on three relatively warm nights, but with fore the antidote was administered. In 18 such cases, the recovery was slow, with 3– speed of 60–80 km/hr. Mean heart rate for all snow leopards was 96 beats per minute male (F5) stood up within 1 min after the had to be physically restrained to receive the antidote both times she was captured.
JOHANSSON ET AL.—REVERSIBLE IMMOBILIZATION OF FREE-RANGING SNOW LEOPARDS standard dose of 80 mg TZ and 0.72 mgMED provided safe immobilization foraverage-size (weight 25–45 kg) adult andsubadult snow leopards. A standard dosethat works on most individuals withouthaving to estimate their weight is useful, asbody weight estimation prior to dartingcan be subjective and made more difficultby the animal’s behavior or capturecircumstances (e.g., in the case of snowleopards lying flat when approached orbody partially hidden behind a rock orother obstruction). The dose used in thisstudy may need to be adjusted for animalsoutside the weight range of the snow rameters of anesthetized free-ranging snow leopards (Panthera uncia). Rectal temperature, heart rate, respiratory rate, and SpO2 (relative oxygen saturationin hemoglobin) were measured during 37 captures in animals with foot-snares is to minimize the Tost Mountains, South Gobi, Mongolia. Data were time the animal spends in the snare.
analyzed using mixed linear models where capture event and individual were treated as random factors.
surveillance system, we observed somemild injuries. However, after we began to was recorded. Capillary refill time never around 1 sec. Muscle relaxation was withinnormal range, and no excessive salivation, Capture-associated lesions and mortalities with lack of responsiveness to tactile stimuli.
leopards continuously after darting, but let sions, small cuts, or between two and four several minutes pass between the observa- fissured claws (mild onychoschisis). In one overestimates, and the true induction times overestimation, induction times were short within 3 mo after any anesthesia event.
immobilized with the same drug combina-tion. Two previous studies report the mean induction time for free-ranging Africanlions to be 14 min (n56) and 5.9 min dine and tiletamine-zolazepam (Fahlman et al., 2005; Jacquier et al., 2006). Captive clinically acceptable physiologic respons- midine and ketamine had similar induction times as the wild snow leopards in our study JOURNAL OF WILDLIFE DISEASES, VOL. 49, NO. 2, APRIL 2013 collars, recording biologic data, and sam- pling blood and hair. Longer duration has been reported in free-ranging lions (Jac- quier et al., 2006; Fahlman et al., 2005) than just ambient temperature. In approx- drug combination, although those investi- ature went below approximately 36.5 C.
(Gulo gulo; Fahlman et al., 2008) and gray duration by providing the antidote as soon wolves (Canis lupus; Ballard et al., 1991).
as possible, thus reducing the anesthetized respiratory rates we observed were ex-pected, and they can be attributed to previous information on respiratory rates ing terrain of the capture site was steep, of free-ranging snow leopards. We record- ed a respiratory rate of 13–16 breaths per leopards in a zoo (Johansson, unpubl.
the relatively high number of spontaneous data). In our study, the anesthetized snow leopards had a substantially higher respi- ratory rate, interpreted as a physiologic response to the elevation in body temper- invariably walked toward steep slopes. In rugged terrain or in areas with open water, an animal that is still under influence of caused an initial mild to moderate tachy- drugs is at risk of injury or mortality.
cardia, which was possibly exacerbated by Therefore, it is desirable to ensure that sedated animals recover in relatively safe (Fahlman et al., 2005). Hence, we believe terrain, away from cliffs or water sources, that the heart rates during anesthesia were although completely avoiding steep terrain above 84%, except for recordings madeduring the first two anesthetic events captures). We believe this was the result of is to be expected due to stress when free- incorrect probe attachment to the animal’s ranging animals are captured. The gradual ear in the first two events, due to the initial JOHANSSON ET AL.—REVERSIBLE IMMOBILIZATION OF FREE-RANGING SNOW LEOPARDS study, logistic constraints such as extreme constant supply of oxygen. The reliability humans as well as animals, and differencesbetween actual arterial values and exter- without the Mongolian Ministry for Nature New, 1983; Hendricks and King, 1993).
and Environment’s cooperation or the gener- ous support of Kolmarden and Nordens Ark.
Henrik Andre´n helped with the statistical trend of recorded values of SpO2 in order Henrik Andre´n, Pernilla Eriksson, and Eva- Charlott Munkenberg helped to improve thismanuscript. We are deeply grateful to Bayar- jargal Agvantseeren and all staff members andvolunteers who assisted at the captures.
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