Eur J Wildl Res (2011) 57:887 893 DOI 10.1007/s10344-011-0500-7 ORIGINAL PAPER Comprison of xylzine ketmine nd medetomidine ketmine nesthesi in the Ierin iex (Cpr pyrenic) Encrn Css-Díz & Ignsi Mrco & Jorge R. López-Olver & Gregorio Menterre & Sntigo Lvín Received: 6 Octoer 2010 / Revised: 13 Jnury 2011 / Accepted: 17 Jnury 2011 / Pulished online: 8 Ferury 2011 # Springer-Verlg 2011 Astrct A comprison ws mde etween two nesthetic comintions in 35 free-rnging dult Ierin iexes (Cpr pyrenic), from My to Decemer 2005. Sixteen iexes (10 mles, 6 femles) were cptured using xylzine ketmine (3.0±0.4+3.0±0.4 mg/kg) nd 19 iexes (12 mles, 7 femles) with medetomidine ketmine (0.10±0.02+2.1±0.3 mg/kg). Anesthetic times were evluted, s well s clinicl vriles (respirtory nd hert rtes, rectl temperture, hemogloin oxygen sturtion), hemtologicl nd iochemicl vriles, t the time of induction nd fter 1 h. The hert rte of iex immoilized with medetomidine ketmine ws higher thn those immoilized with xylzine ketmine. Stiliztion of the hert rte of iex immoilized with medetomidine ketmine cme erlier thn those immoilized with xylzine ketmine. Rectl temperture decresed nd stilized in oth groups, ut erlier in the xylzine ketmine group, nd hypoxemi ws oserved in oth groups. The white lood cell count of iex immoilized with medetomidine ketmine ws lower thn those immoilized with xylzine ketmine throughout nesthesi, while sodium concentrtion ws higher only fter 1 h of nesthesi. In iex immoilized with xylzine ketmine, the neutrophil count, serum cretinine kinse ctivity nd sprtte minotrnsferse ctivity incresed fter 1 h of immoiliztion, while triglycerides decresed. Chnges found in hemtologicl nd iochemicl vriles suggest no Communicted y F. Kup E. Css-Díz (*) : I. Mrco : J. R. López-Olver : G. Menterre : S. Lvín Servei d Ecoptologi de Fun Slvtge (SEFS), Deprtment de Medicin i Cirurgi Animls, Edifici V, Fcultt de Veterinàri, Universitt Autònom de Brcelon, Brcelon, Spin e-mil: encrn.css@u.ct mjor differences in the different drug comintions used, ut clinicl findings of this study, s well s hypoxemi, hypothermi nd rdycrdi, were importnt records tht should e tken into ccount when performing sfe opertion. Keywords Anesthesi. Cpr pyrenic. Ierin iex. Medetomidine. Ketmine. Xylzine Introduction Anesthesi is useful for hndling of species tht re highly sensitive to stress or especilly ggressive. However, it does not completely eliminte or prevent stress or cpture myopthy, possily due to pursuit of the niml efore nd fter the nesthetic drug injection (Hrthoorn 1982; Jessup 1999). For nesthesi in the domestic got, single-drug protocols or comintion of drugs from two nesthetic groups hve een used (Pwde et l. 1996; Afshr et l. 2005; Mhmood nd Mohmmd 2008). The use of single nesthetic drug is very rre in wildlife, it eing more common to ssocite two or more drugs to otin the desired effect. The most frequently used nesthetic comintions in wild ungultes re ketmine xylzine, etorphine cepromcine, tiletmine zolzepm nd medetomidine ketmine (Culkett nd High 2007). Either xylzine or medetomidine, in comintion with ketmine, re used in different ungulte species, such s oudds (Ammotrgus lervi), mouflon (Ovis musimon), xis deer (Cervus xis), fllow deer (Cervus dm), common red deer (Cervus elphus), Pere Dvid s deer (Elphurus dvidinus) nd Alpine iex (Cpr iex iex) (Golightly
888 Eur J Wildl Res (2011) 57:887 893 nd Hofstr 1989; Jlnk nd Roeken 19; Peindo et l. 1999). The Ierin iex (Cpr pyrenic) is used in trnsloctions nd ecologicl studies in Spin, for which its cpture is common prctice. The species hs mrked sexul dimorphism with higher ody weight nd lrger horns in mles, mking hndling difficult nd involving personl risk when using physicl cpture methods. Risks re lower if telenesthesi is used for niml immoiliztion. The ppliction of different nesthetic comintions nd methods provides new, hitherto unpulished, informtion for this species, thus fcilitting the choice of chemicl nesthesi nd method involving the lowest risk for oth humns nd nimls. In this study, two nesthetic comintions xylzine ketmine (XK) nd medetomidine ketmine (MK) were studied to determine differences in nesthetic times, s well s clinicl, hemtologicl nd iochemicl vriles in the free-rnging Ierin iex. Mterils nd methods Thirty-five dult Ierin iexes were cptured in the Sierr Nevd Ntionl Prk, (El Toril estte, 37 03 N, 3 34 O), in Southern Spin, from My to Novemer 2005. The nimls were immoilized y remote drting using CO 2 powered rifle (DAN-Inject, model JM Specil, Børkop, Denmrk) within corrl trp. The nesthetic comintions used were xylzine (3.0±0.4 mg/kg, men ± SD) (Xilgesic 20%, 200 mg/ml, Lortorios Clier, Brcelon, Spin) with ketmine (3.0±0.4 mg/kg, men ± SD) (Imlgène 1000, 100 mg/ml, Meril, Lyon, Frnce) in 16 nimls (10 mles, 6 femles), nd medetomidine (0.10±0.02 mg/kg, men ± SD) (Zlopine, 10 mg/ml, Orion Phrm, Espoo, Finlnd) with ketmine (2.1±0.3 mg/kg, men ± SD) in 19 nimls (12 mles, 7 femles). Administered intrmusculrly ntgonist doses were 1 mg tipmezole (Antisedn, Orion Phrm, Espoo, Finlnd) per 10 mg xylzine, nd 5 mg tipmezole per 1 mg medetomidine. The men ody weight ±SD ws 52.0±11.4 nd 28.9±4.5 kg for mles nd femles, respectively. Five dys were necessry to cpture the nimls (3 dys in My nd 2 dys in Novemer 2005). The Ierin iexes were cptured in corrl trp (100 50 m) ited with food to ttrct them the dy efore the intervention. Once inside, n nesthetic drt with stndrd dose ccording to estimted ody weight ws prepred nd shot with the drt rifle from covered feeder. After induction, the niml ws plced in trnsport sck net, with 4 4 cm mesh (Zioni Ornitecnic, Bergmo, Itly), trnsported outside the enclosure nd weighed. The precordil region ws shved to instll hert rte monitor trnsmitter (Polr Vntge NV nd S710i Polestr, Elect Polestr Oy, Kempele, Finlnd). Ultrsonogrphy gel ws used to ensure good contct etween the electrodes nd the skin (Geleco, Novrtis, Brcelon, Spin). The receiver ws plced in plstic collr round the neck, since it must e less thn 1.5 m from the trnsmitter. In ddition, rectl proes to record temperture were used (Mätmn dtlogger, Chipsoits Eltex AB, Osy, Sweden). The hert rte monitor ws left in plce for 43.9± 11.5 min (men ± SD) nd rectl proes for 40.0±11.2 min (men ± SD). Both vriles were recorded every 60 s, the recorded series eing sved nd turned into numericl vlues with their respective softwre. For sttisticl nlysis, men vlue for every 5 min ws clculted for oth hert rte nd rectl temperture. Stility times were clculted in the hert rte nd temperture. These times were determined compring the vlues long the period of study nd estlishing s time of stiliztion the minute from which differences were not sttisticlly significnt with the following minutes. Respirtory rte nd hemogloin oxygen sturtion ws monitored every 10 min throughout nesthesi. The respirtory rte ws determined y direct oservtion of the costl wll movements; hemogloin oxygen sturtion ws mesured with pulse oximeter with proe ttched to tongue (Vet/Ox 4404, Hesk Corportion, Fort Collins, Colordo, USA). Time of initil effect ws defined s the intervl etween the time of drting nd onset of txi. Time to recumency ws the intervl etween drting nd the time tht the iex ly down in sternl recumency. The time from drting to sfe hndling of nimls ws defined s induction. After 1 h of induction, the ntgonist ws dministered. First rousl ws defined s the point when the niml egn to lift its hed. The time from tipmezole dministrtion up to when the niml returned to stnding position nd multion, with some minor txi, ws defined s recovery. Blood smples were collected from the jugulr vein when the niml ws nesthetized () nd fter 1 h (). Two millilitres were plced in commercil tues contining tripotssium ethylenediminetetrcetic cid (EDTA) s nticogulnt. The remining lood ws plced in tues with polystyrene grnules to fcilitte cogultion nd serum seprtion. The smples were kept in portle refrigertor for processing, lwys within 12 h. Animl cre ctivities nd study procedures were conducted in ccordnce with the guidelines of the good experimentl prctices, under the pprovl of the Ethicl nd Animl Welfre Committee of the Universitt Autònom of Brcelon. Red lood cell count (RBC), white lood cell count (WBC), pltelet count nd hemogloin concentrtion were determined with n electronic impednce semi-utomtic nlyzer (Sysmex F-0; To Medicl Electronics, Hmurg, Germny). Pcked cell volume (PCV) ws mesured y the
Eur J Wildl Res (2011) 57:887 893 889 Tle 1 Men ± SD (min mx) of comprtive immoiliztion nesthetic times of xylzine ketmine nd medetomidine ketmine comintions in Ierin iexes Xylzine+ketmine Medetomidine+ketmine Initil effect (min) 2.6±0.8 (1 4) 5.7±4.7 (2 20) Recumency (min) 5.4±3.2 (1 13) 8.7±5.6 (3 24) Induction (min) 8.2±4.1 (2.5 15) 11.6±7.4 (3.75 30) Antgonist (min) 70.9±4.0 (65 ) 71.3±5.3 (65 81) First rousl (min) 2.7±0.6 (1.72 3) 5.9±4.9 (1.72 20) Recovery (min) 4.6±5.2 (0 20) 5.2±1.4 (2.5 7) stndrd microhemtocrit method, using microhemtocrit centrifuge (Hemtospin 1400, Hwksley, Sussex, UK) t 14,000 G for 6 min. Men corpusculr volume (MCV), men corpusculr hemogloin (MCH) nd men corpusculr hemogloin concentrtion (MCHC) were clculted from the RBC, hemogloin concentrtion nd PCV. The differentil leukocyte count ws mde y identifying 200 leukocytes with n opticl microscope t n originl mgnifiction of 1000, in lood smers dried with pnoptic stin (Chemicl Applied Clinic, Trrgon, Spin). The serum ws otined fter centrifuging the clotted smples t 1,811 G, nd ws frozen t 20 C witing nlysis. Biochemicl vriles such s enzymes [cretine kinse (CK), sprtte minotrnsferse (AST), lnine minotrnsferse (ALT), lkline lctte dehydrogense (LDH), lkline phosphtse (AP)], metolites (ure, glucose, cholesterol, triglycerides, totl iliruin, lctte, cretinine), s well s totl proteins nd chloride, were mesured with two utomtic nlyzers (Cos Mir, Roche, Rotkreuz, Switzerlnd; Olympus AU400, Olympus, Minz, Germny). Sodium nd potssium were mesured y flme photometry (Corning 410 C, Medicl Corning Medfield, USA) nd cortisol levels with n ELISA commercil kit (EIA-1887, DRG Instruments GMBH, Mrurg, Germny). The softwre SAS for System V8 Windows (SAS Inc. Institute, Cry, North Crolin, USA) were used to sttisticl studies, which includes ll procedures pplied. Tests of normlity of the times otined throughout nesthesi were crried out. The non-norml vriles were trnsformed logrithmiclly nd normlity ws nlyzed gin. Vlues regrded s outliers were discrded, cre eing tken not to exceed the mximum llowed y sttisticl studies. An nlysis of vrince (ANOVA) ws pplied to dt considered norml to detect sttisticlly significnt differences etween the groups using the PROC GLM procedure. The nonprmetric dt were nlyzed with the PROC NPAR1WAY ANOVA procedure. The fctor considered in this model for ech vrile ws the nesthetic comintion used; the remining dt (clinicl, hemtologicl nd iochemicl vlues) were nlyzed with n ANOVA, which compred the verges of repeted mesures using the PROC MIXED procedure. The min fctor ws the tretment used, with either of the two nesthetic comintions, nd the repeted fctor ws time. Lest squre mens (LS MEANS) ws used due to the unlnced distriution of nimls etween groups. In ll cses, the ccepted level of significnce ws p<0.05. Results Anesthesi nd recovery times re reported in Tle 1. Some of the iexes presented pnes up to 30 s. The hert rte remined stle nd the temperture decresed over time in oth groups (Fig. 1). The hert rte stilized erlier in the group nesthetized with MK (40 min) thn in the Hert rte (ets/min) Temperture ( o C) 70 60 50 40 30 39.5 39.0 38.5 38.0 37.5 37.0 36.5 36.0 25-45 min 5 10 15 20 25 30 35 40 45 50 55 Time (minutes) 5 10 15 20 25 30 35 40 45 50 55 Time (minutes) Fig. 1 Hert rte nd temperture (men vlues) in Ierin iex immoilized with xylzine ketmine or medetomidine ketmine. Verticl rrows indicte stility times; minutes in ox indicte significnt (p<0.05) differences etween nesthetic comintions
8 Eur J Wildl Res (2011) 57:887 893 Tle 2 Minimum nd mximum vlues for the hert rte nd the rectl temperture 26.0 Hert rte Temperture 25.5 Xylzine+ketmine 39.6 104.8 33.0 40.9 Medetomidine+ketmine 40.4 114.0 35.1 42.8 XK group (45 min), nd decresed nd remined lower for the lst 20 min of the study in nimls immoilized with MK. The temperture stilized erlier in the group nesthetized with XK (25 min) thn in the MK group (40 min) (Fig. 1). Minimum nd mximum vlues for the hert rte nd the rectl temperture re shown in Tle 2. Tle 2 nd chnges in respirtory rte nd hemogloin oxigen sturtion in Figure 2. The MCV decresed significntly over time in the MK group, nd the leukocyte counts ws lower thn in the XK group throughout the study. The neutrophil counts incresed significntly over time in the group nesthetized with XK (Fig. 3). Triglycerides decresed, nd cretine kinse (CK) nd sprtte minotrnsferse (AST) incresed over time in the XK group. Sodium concentrtion ws higher in the group nesthetized with MK t the end of the study s compred with the XK group (Fig. 4). 130 120 110 100 70 60 100 95 85 75 70 Respirtory rte (reths per minute) 20 min 30 min 40 min 50 min Hemogloin oxygen sturtion (%) 20 min 30 min 40 min 50 min Fig. 2 Respirtory rte nd hemogloin oxygen sturtion in Ierin iex immoilized with xylzine ketmine or medetomidine ketmine MCV (fl) WBC (x10 9 /l) c Neutrophils (x10 9 /l) 25.0 24.5 24.0 23.5 23.0 16 15 14 13 12 11 10 9 8 7.0 6.5 6.0 5.5 5.0 4.5 4.0 Discussion Anesthetic gents reported for use in Ierin iex include etorphine (Pérez et l. 1999), tiletmine zolzepm (Peindo et l. 1993, Sntigo-Moreno et l. 2011), detomidine ketmine (Sntigo-Moreno et l. 2011) nd the XK comintion (Peindo et l. 1991). For similr species, such s Alpine iex (Cpr iex), crfentnil nd xylzine * * Fig. 3 Men corpusculr volume (MCV), white lood cells count (WBC) nd c neutrophils during nesthesi of Ierin iex., mens with different superscript sttisticlly different (p<0.05) from ech other. Asterisks indicte sttisticlly significnt differences (p< 0.05) etween nesthetic comintions
Eur J Wildl Res (2011) 57:887 893 891 (Kreeger nd Arnemo 2002) nd the Hellrunner mix (125 mg xylzin+100 mg ketmin/ml) (Aderhlden et l. 1998) hve lso een used, ut cre must e tken in some species of wild ungultes tht hve specil sensitivity (Wiesner nd von Hegel 1985). Hemtologicl nd serum iochemicl vriles in studies with Ierin iex were similr to those otined in our study, ut few differences should e mentioned. In the study of XK-immoilized iexes, only RBC nd lymphocytes were higher, nd HCM ws lower. The gretest differences re found with iexes immoilized with tiletmine zolzepm (Peindo et l. 1993), which hd lower MCHC, neutrophil count, glucose concentrtion, ure, CK, AST nd LDH, nd higher count of lymphocytes, monocytes nd eosinophils. These differences my e due to pre-cpture hndling (oxtrps previous to nesthesi) nd drugs dministered. Sntigo-Moreno et l. (2011) used comintion of nother lph-2-drenergic gonist (detomidine) nd ketmine, nd otined lower induction nd respirtory rte, lower concentrtion of glucose, cretinine, AST nd CK, nd stle temperture. However, the iexes used in this study were conditioned in snd-floor stle for period of 12 months, nd hnd injection ws fesile. The nimls in our study displyed higher respirtory rte (107±36 reths per minute) thn in the Alpine iex (C. iex, 83±15 reths per minute) immoilized with xylzine (Percino nd Bssno 1993). Also, s in the previous report, the Ierin iex in our study displyed severl pnes lsting up to 30 s. Hypoxemi nd compenstory tchypne to lnce the lck of oxygen could ccount for this difference. However, in ruminnts, in ddition to hypoventiltion during nesthesi, intrpulmonry cuses (such s ventiltion-perfusion mismtch, shunt nd diffusion impirment) nd recumency cn cuse hypoxemi, which my effectively e treted with supplementl oxygen (Admson nd Mills 1996). Both xylzine nd medetomidine cn cuse hypotension nd rdycrdi, ut medetomidine hs ten times more specificity for lph-2 receptors versus lph-1 receptors when compred to xylzine (Gross nd Booth 1995; Plum CK (IU/l) 0 750 700 650 600 550 500 450 400 350 300 AST (UI/l) 160 150 140 130 120 110 100 c Triglycerides (mmol/l) 0.12 0.11 0.10 0.09 0.08 0.07 0.06 0.05 0.04 d Sodium (mmol/l) 154 152 150 148 146 144 142 140 * Fig. 4 Serum cretine kinse (CK), sprtte minotrnsferse (AST), c triglycerides nd d sodium during nesthesi of Ierin iex., mens with different superscript sttisticlly different (p< 0.05) from ech other. Asterisks indicte sttisticlly significnt differences (p<0.05) etween nesthetic comintions
892 Eur J Wildl Res (2011) 57:887 893 2008). In fct, Celly et l. (1997) found tht xylzine did not ffect the hert rte in sheep. In ddition, Jlnk nd Roeken (19) found tht medetomidine produces hypothermi in non-domestic mmmls. There re no reference vlues for sl (resting) clinicl prmeters in the Ierin iex. But, if we compre our results with the reference vlues given for domestic gots (Jckson nd Cockcroft 2002), the hert rte nd the ody temperture of the Ierin iex nesthetized with MK were lower, which could indicte rdycrdi nd hypothermi. Sntigo-Moreno et l. (2011) found lso rdycrdi in iexes treted with detomidine. Chnges found in hemtologicl nd iochemicl vriles in the Ierin iex of our study suggest no mjor differences etween the two drug comintions used. The decrese in the MCV in the group of MK nd the increse in neutrophils nd higher WBC in the group of XK is unlikely due to stress within treted groups ecuse other chnges in hemtologicl vriles such s RBC, PCV, concentrtion of hemogloin nd lymphocytes should hve een expected. The increse in CK nd AST in the XK group could e due to muscle necrosis cused y musculr injection or musculr contrctions (Gertner et l. 1987, Bouts et l. 2010). Also, the decrese of triglycerides nd the lower concentrtion of sodium in the XKimmoilized iex re difficult to relte with different response to cpture stress. These vriles my not e useful s stress indictors, since some uthors hve found chnges wheres others hve not (Kocn et l. 1981; Broom nd Johnson 1993; Peindo et l. 1993; Montné et l. 2003). Proly, other fctors not considered in our study, such s seson, nutrition nd environmentl temperture, my ffect some of these vriles, nd it would e needed to e further nlyzed. 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