Veterinary Anaesthesia and Analgesia, 2016 doi:10.1111/vaa.

12339

RESEARCH PAPER

Behavioral and cardiopulmonary effects of

dexmedetomidine alone and in combination with
butorphanol, methadone, morphine or tramadol in
conscious sheep

Luisa PB Borges*, Lilian T Nishimura*, Leonardo L Carvalho*, Sofia A Cerejo†, Adam Auckburally‡ &
Ewaldo Mattos-Junior*
*School of Veterinary Medicine, University of Franca, Franca, Brazil
†School of Veterinary Medicine and Animal Science, UNESP-Botucatu, Botucatu, Brazil
‡School of Veterinary Medicine, College of Medical Veterinary & Life Sciences, University of Glasgow, Glasgow, Scotland

Correspondence: Ewaldo de Mattos Junior, Faculdade de Medicina Veterin

aria, Universidade de Franca, Avenida Dr Armando Salles de
Oliveira, 201, Parque Universit
ario, Franca, SP, Brazil, 14404-600. E-mail: ewaldomattos@hotmail.com

Results HR decreased in all treatments and fR

Abstract
Objective To compare cardiopulmonary and seda-
tive effects following administration of dexmedeto-
midine alone or with butorphanol, methadone,
morphine or tramadol in healthy sheep.
Study design Randomized crossover study.
Animals Six Santa In^es sheep, five females, one
male, aged 12–28 months and weighing
40.1  6.2 kg.
decreased in DM at T30 and DMO at T30 and T45.
PaCO2 was increased in D, DB and DM compared
with baseline, and PaO2 decreased in D at T15 and
T45; in DB at T15 to T75; in DM at T15 to T60; in
DMO at T15; and in DT at T15, T30 and T75. There
was a decrease in temperature in D, DB and DM. An
increased pH was measured in D at all time points
and in DT at T30–T120. HCO3 and base excess were
increased in all treatments compared with baseline.
There were no statistical differences in sedation
scores.

Methods Sheep were assigned treatments of
dexmedetomidine (0.005 mg kg 1; D); D and
butorphanol (0.15 mg kg 1; DB); D and metha-
done (0.5 mg kg 1; DM); D and morphine (0.5 mg
kg 1; DMO); or D and tramadol (5.0 mg kg 1; DT).
All drugs were administered intravenously with at
least 7 days between each treatment. Rectal
temperature, heart rate (HR), respiratory rate (fR),
invasive arterial pressure, blood gases and elec-
trolytes were measured prior to administration of
drugs (baseline, T0) and every 15 minutes follow-
ing drug administration for 120 minutes (T15–
T120). Sedation was scored by three observers
blinded to treatment.
Conclusions and clinical relevance The combina-
tion of dexmedetomidine with butorphanol, metha-
done, morphine or tramadol resulted in similar
changes in cardiopulmonary function and did not
improve sedation when compared with dexmedeto-
midine alone.
Keywords a2-agonists, cardiopulmonary, opioids,
ovine.
Introduction
Alpha2-adrenergic agonists are used for sedation
and premedication prior to general anesthesia in

1
Dexmedetomidine–opioids in sheep LPB Borges et al.
several species. Racemic medetomidine has a bind-
ing ratio of 1620:1 (a2:a1) (Virtanen et al. 1988)
and its d-enantiomer, dexmedetomidine, is even
more selective (Murrell & Hellebrekers 2005).
Advantages of a2-agonists include potent, pre-
dictable sedation (Cardoso et al. 2014), analgesia,
reduced anesthetic requirement, and reversibility
(Murrell & Hellebrekers 2005).
In sheep, a2-agonists are widely used for provi-
sion of analgesia and sedation (K€ astner 2006).
However, arterial hypoxemia and pulmonary
edema have been reported in certain breeds of
sheep following the administration of all a2-
agonists, including dexmedetomidine (Celly et al.
1997; K€astner et al. 2001b; K€astner 2006).
Congestion and redistribution of blood flow have
been suggested as the cause of impaired oxygena-
tion following the administration of dexmedeto-
midine to healthy anesthetized sheep. The
hypoxemia is made worse by alveolar edema as a
result of hydrostatic stress (K€ astner et al. 2007).
Dexmedetomidine has been compared with medeto-
midine in sheep, and has similar cardiopulmonary
and sedative effects (K€astner et al. 2001a), but the
effects of combinations of dexmedetomidine and
opioids have not yet been described.
The administration of dexmedetomidine with
opioids to dogs (Cardoso et al. 2014), and of xylazine
with opioids to sheep (de Carvalho et al. 2015),
improves sedation when compared with administra-
tion of the a2-agonist alone. Combining dexmedeto-
midine with opioids in conscious sheep may facilitate
certain procedures, and lower doses might reduce
the incidence and severity of side effects.
The aim of this study was to compare the
cardiopulmonary and sedative effects of dexmedeto-
midine alone or in combination with butorphanol,
methadone, morphine or tramadol in sheep. Our
hypothesis was that these combinations may
improve sedation without inducing significant car-
diopulmonary depression when compared with the
administration of dexmedetomidine alone.
Materials and methods
This research was conducted following approval
from the Animal Ethics Committee of the University
of Franca, SP, Brazil (038/12). The research facility
is located 1040 m above sea level. Detailed infor-
mation regarding the management and assessment
of animals prior to experimentation, and also for
further details of measurement methods, can be
2 © 2016 Association of Veterinary Anaesthetists and the American College of Veterinary Anesthesia and Analgesia
found in a previous associated study (de Carvalho
et al. 2015).
Animals
Six Santa In^es sheep, five females and one male, aged
12–28 months and weighing 40.1  6.2 kg, were
used. Catheters were inserted aseptically into a
jugular vein (18 gauge, 2.5 cm) and an auricular
artery (20 gauge, 2.5 cm) with the sheep standing.
Variables were measured prior to the administration
of drugs (baseline, T0) and then every 15 minutes
following the administration of drugs for 120 min-
utes (T15–T120).
Experimental design
Sheep were administered treatments in random order
(by drawing numbered papers from a box) in a
crossover design with a washout period of 7 days
between treatments. The treatments were as
follows: D, dexmedetomidine (0.005 mg kg 1; Dex-
domitor, 0.5 mg mL 1; Pfizer, UK); DB, dexmedeto-
midine (0.005 mg kg 1) and butorphanol
(0.1 mg kg 1; Torbugesic, 10 mg mL 1; Fort
Dodge, IA, USA); DM, dexmedetomidine
(0.005 mg kg 1) and methadone (0.5 mg kg 1;
Mytadon, 10 mg mL 1; Crist
alia Produtos Qu
ımicos
e Farmac^euticos Ltda, SP, Brazil); DMO, dexmedeto-
midine (0.005 mg kg 1) and morphine
(0.5 mg kg 1; Dimorf, 10 mg mL 1; Crist
alia Pro-
dutos Qu
ımicos e Farmac^euticos Ltda); or DT,
dexmedetomidine (0.005 mg kg 1) and tramadol
(5.0 mg kg 1; Tramadon, 50 mg mL 1; Crist
alia
Produtos Qu
ımicos e Farmac^euticos Ltda). After
instrumentation, a 15 minute period of stabilization
was allowed prior to data collection. All drugs
administered were mixed in the same syringe with
the final volume adjusted to 10 mL with 0.9% sodium
chloride to facilitate blinding and given intravenously
(IV) over 30 seconds into the jugular catheter.
Degree of sedation
The degree of sedation was assessed using a numer-
ical rating scale of 0–10, as follows: 0, no sedation;
1, standing, alert, reduced head and ear movements;
2, standing, slight head drop; 3, standing, moderate
head drop; 4, standing, severe head drop, ataxia; 5,
standing, severe head drop, severe ataxia; 6, sternal
recumbency, head up; 7, sternal recumbency, head
down; 8, lateral recumbency, occasional attempts to

attain sternal recumbency; 9, lateral recumbency,
uncoordinated movements; and 10, lateral recum-
bency, no movements (K€ astner et al. 2003; de
Carvalho et al. 2015).
Cardiopulmonary variables and rectal temperature
Heart rate (HR) was counted by thoracic ausculta-
tion with a stethoscope and respiratory rate (fR) by
observation of thoracic excursions, each over
1 minute. Mean arterial pressure (MAP) was mea-
sured from an auricular artery catheter connected to
an aneroid manometer (Ind
ustria Bic de Aperelhos
M
edicos Ltda, SP, Brazil), which was calibrated
against a mercury column prior to use, and tubing
filled with 0.1% heparin solution (50 IU mL 1) and
the air–saline junction was aligned with the point of
the shoulder in standing and sternally recumbent
animals and the xiphoid process in laterally recum-
bent animals (de Carvalho et al. 2015). Hypotension
was defined as MAP <60 mmHg. Rectal temperature
(RT) was measured with a mercury-in-glass ther-
mometer (Thermometer BD; Becton Dickinson
Ind
ustrias Cirurgicas SA, MG, Brazil).
Blood gases and electrolytes
Arterial blood samples were collected for determi-
nation of pH, partial pressure of carbon dioxide
(PaCO2), partial pressure of oxygen (PaO2), base
excess (BE), arterial hemoglobin oxygen saturation
(SaO2), bicarbonate (HCO3 ), sodium (Na+), potas-
sium (K+) and chloride (Cl ) concentrations. After
first withdrawing 2 mL of blood from the arterial
catheter, 0.5 mL of blood for analysis was drawn
into a disposable heparinized syringe and sealed
with a rubber stopper. Blood samples were analyzed
immediately [Cobas b 121; Roche Diagnostics
(Schweiz) AG, Switzerland]. Hypoxemia was defined
as PaO2 <60 mmHg.
Statistical analysis
The results were analyzed using a statistical analysis
software program GraphPad PRISM Version 5.0
(GraphPad Software, Inc., CA, USA). Normality was
assessed using the Shapiro–Wilk test. Normally
distributed data were analyzed using analysis of
variance (ANOVA) for repeated measures. Post hoc
analysis within the same treatment was performed
using Dunnett’s test and between treatments using
Bonferroni correction. Nonparametric data were
© 2016 Association of Veterinary Anaesthetists and the American College of Veterinary Anesthesia and Analgesia
Dexmedetomidine–opioids in sheep LPB Borges et al.
analyzed using the Friedman test followed by post
hoc Dunn’s test. For all data p < 0.05 was considered
to be significant.
Results
All animals completed the 120 minutes of evalua-
tion. Behavioral effects other than sedation included
salivation, mydriasis, bruxism (teeth grinding),
vocalization and facial muscle tremors (Table 1).
The sheep recovered from sedation without further
complications.
Sedative effects
Sedation scores were significantly higher compared
with baseline at T15–T60 in D and DT; at T15–T75
in DB and DM; and at T15–T90 in DMO (Fig. 1).
There was no significant difference in the compar-
ative analysis among treatments. Sternal or lateral
recumbency (scores 6–10) occurred in D at six time
points (T15–T90), DB and DM at four time points
(T15–T60) and DMO at five time points (T15–T75).
Recumbency did not occur in any animal in DT
(Fig. 1).
Cardiopulmonary variables and RT
There was a significant reduction in HR at all time
points compared with baseline in D, DB and DT; in
DM at T45, T75 and T105; and in DMO at T15–T60
(Table 2). There were no significant differences
among treatments. With the exception of T105 in
DT, MAP did not change significantly from baseline
in any treatment, and there were no significant
differences among the treatments. Significant
decreases were measured in fR compared with
baseline in DM at T30 and in DMO at T30–T60.
There were no significant differences among treat-
ments (Table 2). Temperature decreased signifi-
cantly from baseline in D at T60 and T75, in DB at
T45–T120, and in DM at T45, T75 and T90
(Table 2). There were no significant differences in
RT among the treatments.
Blood gas and electrolyte analysis
Mean pH values were higher compared with baseline
in D at all time points, in DB at T90–T120, and in DT
at T60–T120 (Table 3). There was no significant
difference in pH among treatments. There was a
significant increase in PaCO2 compared with base-
3
 4
Table 1 Adverse effects observed in six sheep following intravenous administration of dexmedetomidine (0.005 mg kg 1) (D), dexmedetomidine and butorphanol (0.15 mg kg 1) (DB),
dexmedetomidine and methadone (0.5 mg kg 1) (DM), dexmedetomidine and morphine (0.5 mg kg 1) (DMO), or dexmedetomidine and tramadol (5 mg kg 1) (DT) observed over
120 minutes after drug administration

Time points (minutes)

Treatment T15 T30 T45 T60 T75 T90 T105 T120

D Mydriasis, urination Mydriasis, drooling Mydriasis, urination, Drooling, urination, None None None None
vocalization vocalization,
Dexmedetomidine–opioids in sheep LPB Borges et al.

bruxism
DB Recumbency with Drooling, urination, Drooling, urination, Drooling, urination, Drooling, Drooling, vocalization Drooling, vocalization None
paddling, urination mydriasis mydriasis, mydriasis, vocalization,
nystagmus mydriasis,
muscular tremors
DM Drooling, bruxism, Bruxism, drooling, Bruxism, mydriasis, Bruxism, mydriasis, Bruxism, urination Bruxism, vocalization Bruxism, vocalization Bruxism,
facial tremors vocalization, drooling, vocalization, vocalization
urination vocalization, drooling
muscular tremors
DMO Bruxism, facial Drooling, bruxism, Drooling, urination, Vocalization, Vocalization, Mydriasis None None
tremors, urination, mydriasis, bruxism muscular tremors, muscular tremors,
vocalization mydriasis, nystagmus, mydriasis
mydriasis
DT Urination, Mydriasis, drooling, Mydriasis, urination, Mydriasis, urination, Mydriasis, urination, Mydriasis, Mydriasis, Mydriasis
vocalization, muscular tremors drooling, drooling drooling vocalization vocalization
drooling, severe vocalization
facial tremors

© 2016 Association of Veterinary Anaesthetists and the American College of Veterinary Anesthesia and Analgesia
 Dexmedetomidine–opioids in sheep LPB Borges et al.

Figure 1 Sedation scores, median (square symbol) and range (circles), in six sheep before (time 0) and after intravenous
injection of: (a) dexmedetomidine (0.005 mg kg 1) (D); (b) dexmedetomidine (0.005 mg kg 1) and butorphanol
(0.15 mg kg 1) (DB); (c) dexmedetomidine (0.005 mg kg 1) and methadone (0.5 mg kg 1) (DM); (d) dexmedetomidine
(0.005 mg kg 1) and morphine (0.5 mg kg 1) (DMO); and (e) dexmedetomidine (0.005 mg kg 1) and tramadol
(5 mg kg 1) (DT). Scores below the dotted line indicate the animal was standing, scores between the dotted and solid
lines indicate that animals were in sternal recumbency and those above the solid line indicate that animals were in lateral
recumbency. *Significantly different from baseline within the same treatment (p < 0.05).

line at all time points in D and DB, and in DM at
T15–T90, with no difference among treatments
(Table 3).
There was a significant increase in [HCO3 ] com-
pared with baseline in D, DB and DM at T15–T120;
in DMO at T15–T105; and in DT at T30–T120 min-
utes. Base excess was significantly increased com-
pared with baseline in D at T45–T120 minutes, in
DB at all time points, in DM at T30–T90, in DMO at
T45 and T60 and in DT at T30–T120 (Table 3).
There were no significant differences among treat-
ments in BE and [HCO3 ].
There was a significant decrease in PaO2 com-
pared with baseline in D at T15 and T45, in DB at

© 2016 Association of Veterinary Anaesthetists and the American College of Veterinary Anesthesia and Analgesia 5
 6
Table 2 Heart rate (HR), mean arterial pressure (MAP), respiration rate (fR) and rectal temperature (RT) in six sheep before (T0) and after intravenous injection of dexmedetomidine (D),
dexmedetomidine–butorphanol (DB), dexmedetomidine–methadone (DM), dexmedetomidine–morphine (DMO) or dexmedetomidine–tramadol (DT)

Time points (minutes)

Variable Treatment T0 T15 T30 T45 T60 T75 T90 T105 T120

HR (beats minute 1) D 101  13 71  16‡ 72  18‡ 72  17‡ 65  13‡ 67  7‡ 75  20‡ 73  13‡ 74  8‡

DB 105  28 75  18‡ 74  20‡ 70  15‡ 59  9‡ 63  5‡ 67  11‡ 66  12‡ 64  8‡
DM 99  11 79  13 80  24 67  18* 70  15 66  15* 71  15 68  15* 68  11
DMO 99 20 67 12‡ 65 16‡ 69 22‡ 68 19‡ 83 17 87 21 85 20 89 15
Dexmedetomidine–opioids in sheep LPB Borges et al.

        
DT 80  13 61  8‡ 59  8‡ 59  5‡ 57  3‡ 59  3‡ 63  8‡ 66  9‡ 64  4‡
MAP (mmHg) D 101  14 96  8 95  9 90  8 92  9 90  5 100  14 99  10 96  11
DB 105  6 96  21 92  20 98  23 95  10 96  8 90  7 92  7 92  8
DM 108  10 104  9 99  12 98  10 97  13 94  14 103  19 101  17 97  13
DMO 104  9 92  8 96  13 99  10 95  9 101  13 101  13 99  11 101  10
DT 111  8 101  17 105  9 104  15 108  15 99  6 95  13 93  7* 98  8
fR (breaths minute 1) D 43  11 36  14 35  15 37  27 34  19 35  22 37  22 35  21 33  20
DB 28  10 34  18 23  9 20  4 27  9 23  4 17  5 19  4 18  3
DM 27  12 20  5 17  4* 21  7 20  5 21  5 21  7 21  7 23  10
DMO 35  11 27  11 26  11* 27  15* 27  9* 29  13 31  17 34  18 28  15
DT 30  10 34  14 34  12 26  8 30  21 23  6 25  12 25  9 24  6
RT (°C) D 39.3  0.3 39.3  0.4 39.0  0.4 39.0  0.3 38.8  0.3* 38.9  0.4* 39.0  0.3 39.0  0.4 39.1  0.3
DB 39.0  0.5 38.7  0.6 38.6  1.0 38.5  1.1* 38.2  0.7‡ 38.2  1.0‡ 38.3  0.9‡ 38.2  1.1‡ 38.4  1.0‡
DM 39.1  0.6 38.8  0.6 38.6  0.6 38.4  0.7* 38.5  0.9 38.4  1.0* 38.4  0.9* 38.5  1.0 38.5  1.0
DMO 39.4  0.7 39.4  0.5 39.3  0.6 39.2  0.8 39.0  0.7 39.0  1.0 39.1  0.7 39.2  0.4 39.2  0.5
DT 38.9  0.7 38.8  0.7 38.9  0.5 38.7  0.6 38.6  0.7 38.7  0.8 38.8  0.7 38.7  0.8 38.8  0.8

Data are means  standard deviation. *(p < 0.05), ‡(p < 0.001), significantly different from T0 within the same treatment.

© 2016 Association of Veterinary Anaesthetists and the American College of Veterinary Anesthesia and Analgesia
 Table 3 Arterial pH, partial pressure of carbon dioxide (PaCO2), partial pressure of oxygen (PaO2), bicarbonate (HCO3 ), base excess (BE) and arterial hemoglobin oxygen saturation
(SaO2) in six sheep before (T0) and after intravenous injection of dexmedetomidine (D), dexmedetomidine–butorphanol (DB), dexmedetomidine–methadone (DM), dexmedetomidine–
morphine (DMO) or dexmedetomidine–tramadol (DT)

Time points (minutes)

Variable/treatment T0 T15 T30 T45 T60 T75 T90 T105 T120

pH
D 7.47  0.04 7.49  0.03† 7.51  0.01† 7.51  0.02† 7.52  0.01† 7.52  0.02† 7.52  0.04† 7.52  0.02† 7.52  0.02†
DB 7.45  0.10 7.45  0.08 7.47  0.06 7.48  0.06 7.48  0.09 7.51  0.05 7.51  0.05* 7.51  0.05* 7.52  0.04*
DM 7.51  0.02 7.46  0.03 7.46  0.03 7.48  0.02 7.51  0.03 7.51  0.02 7.42  0.19 7.51  0.06 7.50  0.05
DMO 7.47  0.07 7.48  0.06 7.50  0.04 7.51  0.03 7.51  0.04 7.52  0.03 7.51  0.04 7.51  0.03 7.52  0.04
DT 7.48  0.02 7.49  0.02 7.50  0.02 7.50  0.02 7.51  0.01* 7.52  0.01† 7.53  0.01‡ 7.54  0.03‡ 7.52  0.03‡
PaCO2 (mmHg)
D 27  4 33  2‡ 33  3‡ 33  3‡ 34  2‡ 34  2‡ 33  4‡ 32  2† 33  2‡
DB 28  5 35  6‡ 36  5‡ 37  6‡ 37  4‡ 35  5‡ 35  5‡ 35  6‡ 35  6‡
DM 30  3 39  6‡ 39  6‡ 38  7‡ 36  6† 36  4† 35  4* 33  4 34  3
DMO 30  2 35  3 36  3 36  3 38  6* 32  6 32  6 33  3 32  3
DT 30  2 34  2 34  2 35  2 34  2 34  4 34  2 31  8 34  3
PaCO2 (kPa)
D 3.5  0.5 4.3  0.3‡ 4.3  0.4‡ 4.3  0.4‡ 4.4  0.3‡ 4.4  0.3‡ 4.3  0.5‡ 4.1  0.3† 4.3  0.3‡
DB 3.6  0.6 4.6  0.8‡ 4.7  0.6‡ 4.8  0.8‡ 4.8  0.5‡ 4.5  0.6‡ 4.5  0.6‡ 4.5  0.8‡ 4.5  0.8‡
DM 4.0  0.4 5.0  0.8‡ 5.0  0.8‡ 4.9  0.9‡ 4.7  0.8‡ 4.7  0.5† 4.5  0.5* 4.3  0.5 4.4  0.4
DMO 4.0  0.3 4.5  0.4 4.7  0.4 4.7  0.4 4.9  0.8* 4.1  0.8 4.1  0.8 4.3  0.4 4.1  0.4
DT 4.0  0.3 4.4  0.3 4.4  0.3 4.5  0.3 4.4  0.8 4.4  0.5 4.4  0.3 4.0  1.0 4.4  0.4
PaO2 (mmHg)
D 80  7 68  10‡ 73  12 70  8* 74  6 73  6 72  10 76  5 78  5
DB 85  9 64  15‡ 67  13† 69  8† 70  6‡ 73  3* 77  7 76  5 73  4
DM 83  5 60  15‡ 67  13† 70  13* 69  10* 77  5 77  6 77  6 79  6
DMO 83  4 68  6* 70  6 74  6 72  4 82  15 82  15 81  17 79  5
DT 83  7 73  3* 74  3* 76  6 75  3 74  6 76  4 75  5 77  9

© 2016 Association of Veterinary Anaesthetists and the American College of Veterinary Anesthesia and Analgesia
PaO2 (kPa)
D 10.4  0.9 8.8  1.3‡ 9.5  1.6 9.1  1.0* 9.6  0.8 9.5  0.8 9.3  1.3 9.8  0.6 10.1  0.6
DB 11.0  1.0 8.3  1.9‡ 8.7  1.7† 9.0  1.0† 9.1  0.8‡ 9.5  0.4* 10.0  0.9 9.8  0.6 9.5  0.5
DM 10.8  0.6 7.8  1.9‡ 8.7  1.7† 9.1  1.7* 9.0  1.3* 10.0  0.6 10.0  0.8 10.0  0.8 10.2  0.8
DMO 10.8  0.5 8.8  0.8* 9.1  0.8 9.6  0.8 9.3  0.5 10.6  1.9 10.6  1.9 10.5  2.2 10.2  0.6
DT 10.8  0.9 9.4  0.4* 9.6  0.4* 9.9  0.8 9.7  0.4 9.6  0.8 9.9  0.5 9.7  0.6 10.0  1.1

7
Dexmedetomidine–opioids in sheep LPB Borges et al.

(continued)
 8
Table 3 (continued)

Time points (minutes)

Variable/treatment T0 T15 T30 T45 T60 T75 T90 T105 T120

HCO3 (mmol L 1)
D 20  4 24  2† 27  2‡ 26  2‡ 27  1‡ 28  2‡ 24  7‡ 26  2‡ 26  2‡
DB 20  7 25  7‡ 26  7‡ 27  7‡ 27  6‡ 28  5‡ 28  5‡ 28  6‡ 28  6‡
DM 23  3 27  3‡ 27  4‡ 28  4‡ 28  3‡ 28  3‡ 27  5‡ 26  4* 26  4†
Dexmedetomidine–opioids in sheep LPB Borges et al.

DMO 23  2 26  3† 27  2† 28  3† 29  2† 25  5† 25  5† 25  4* 24  2
DT 22  2 25  2 26  2* 27  1‡ 27  2‡ 27  3‡ 27  2‡ 26  6‡ 27  3‡
BE (mmol L 1)
D –1  5 1  2 3  2 3  2† 4  1† 4  2† 3  3* 3  2* 4  2*
DB 4  0 4  3* 5  4‡ 5  5‡ 6  4‡ 5  5‡ 5  5‡ 5  6‡ 5  6‡
DM 1  3 3  3 3  3* 4  3† 5  3‡ 5  2‡ 4  5† 3  4 3  4
DMO 1  2 3  3 4  2 5  2* 5  3* 2  5 1  6 1  5 2  2
DT 0  2 2  1 3  2* 4  1† 4  2‡ 5  2‡ 5  2‡ 4  4‡ 4  3‡
SaO2 (%)
D 96  1 94  2* 96  2 96  1 96  1 96  1 96  1 97  1 97  0
DB 97  1 91  6* 94  3 95  1 96  2 96  1 97  1 96  1 96  1
DM 97  0 89  8*§ 93  5 95  2 95  2 97  1 97  1 97  1 97  0
DMO 97  1 95  2* 95  1* 95  1 96  2 96  1 97  1 97  1 97  1
DT 97  1 96  1 96  1 96  1 96  1 96  1 97  1 97  1 97  1

Data are means  standard deviation. *(p < 0.05), †(p < 0.01), ‡(p < 0.001), significantly different from T0 within the same treatment. §Significantly different from other treatments at the same time point
(p < 0.05).

© 2016 Association of Veterinary Anaesthetists and the American College of Veterinary Anesthesia and Analgesia
 Dexmedetomidine–opioids in sheep LPB Borges et al.

T15–T75, in DM at T15–T60, in DMO at T15 and in
DT at T15 and T30. There were no significant
differences among treatments. Arterial oxygen sat-
uration was significantly lower at T15 compared
with baseline in D, DB, DM and in DMO at time
points T15 and T30. SaO2 was significantly lower in
DM at T15 compared with other treatments.
Plasma sodium concentration was significantly
increased compared with baseline in DMO at T105
and in DT at T90–T120 (Table 4). There was no
significant difference among treatments. [K+] was
significantly lower compared with baseline in DMO
and DT at T90–T120, and significantly higher in DB
compared with other treatments at T120 minutes.
Chloride concentration was significantly lower com-
pared with baseline in DB at T15 and T30 (Table 4).
There was no significant difference among treat-
ments.
Discussion
Dexmedetomidine has been used in sheep as premed-
ication prior to general anesthesia (K€astner et al.
2001 a, b, 2007; K€ astner 2006; Granados et al.
2012; Funes et al. 2014). Doses administered ranged
from 0.0025 to 0.015 mg kg 1 in these studies.
Concurrent administration of dexmedetomidine and
an opioid in dogs results in significantly enhanced
sedation without additional cardiopulmonary side
effects (Cardoso et al. 2014). A relatively low dose of
dexmedetomidine (0.005 mg kg 1) was chosen for
this study in sheep, as it was to be combined with a
variety of opioids. A possible explanation for findings
that differed in comparison with an associated
experiment studying combinations of xylazine with
opioids (de Carvalho et al. 2015) could be that the
characteristics of the two drugs are different, in that
dexmedetomidine induces more sedative effect than
xylazine; consequently, the effects of the drug-opioid
combinations were compared with different degrees
of sedation at baseline. The sheep administered
dexmedetomidine alone in this present study
achieved higher mean sedation scores with recum-
bency than sheep administered xylazine alone, so
that an additional sedative effect of the opioid might
not have been as obvious.
Equipotent doses of opioids in sheep are not
reported and, therefore, the dose rates chosen for
this study were based on studies performed in dogs
(Mastrocinque & Fantoni 2003; Maiante et al.

Table 4 Plasma sodium (Na+), potassium (K+) and chloride (Cl ) in six sheep before (T0) and after intravenous injection of
dexmedetomidine (D), dexmedetomidine–butorphanol (DB), dexmedetomidine–methadone (DM), dexmedetomidine–mor-
phine (DMO) or dexmedetomidine–tramadol (DT)

Time points (minutes)

Variable/
treatment T0 T15 T30 T45 T60 T75 T90 T105 T120

Na+ (mmol L 1)
D 148  2 147  3 147  2 147  2 147  2 147  2 147  2 150  5 147  1
DB 147  3 145  3 145  2 145  1 146  1 146  2 147  1 147  2 146  1
DM 145  4 146  2 145  1 144  2 145  3 146  2 145  2 147  3 146  2
DMO 143  2 145  2 145  1 145  1 145  3 147  6 147  5 148  3* 146  2
DT 145  2 144  2 146  3 146  2 146  2 147  3 148  3* 150  4* 149  4*
K+ (mmol L 1)
D 3.1  0.4 3.0  0.3 3.1  0.1 3.0  0.2 3.0  0.3 3.1  0.3 3.2  0.4 3.0  0.2 3.1  0.2
DB 3.3  0.2 3.3  0.3 3.4  0.4 3.4  0.5 3.2  0.4 3.3  0.6 3.3  0.4 3.7  0.8 4.5  2.7§
DM 3.1  0.4 3.0  0.3 3.1  0.3 3.1  0.3 3.0  0.5 3.0  0.4 3.1  0.4 2.9  0.4 3.0  0.3
DMO 3.5  0.1 3.3  0.2 3.3  0.2 3.3  0.3 3.3  0.3 3.2  0.4 3.2  0.4* 3.1  0.4* 3.1  0.3*
DT 3.6  0.4 3.5  0.4 3.5  0.4 3.4  0.3 3.4  0.3 3.3  0.2 3.2  0.3* 3  0.5* 3.1  0.3*
Cl (mmol L 1)
D 110  2 111  12 108  4 106  2 106  3 105  1 107  1 108  4 106  2
DB 109  5 104  5* 104  4* 105  4 106  2 105  2 106  2 105  3 106  3
DM 105  2 105  2 103  2 103  2 104  2 103  1 103  1 105  4 103  2
DMO 105  3 107  3 106  3 104  3 105  3 108  7 108  6 107  3 106  4
DT 106  2 104  1 106  4 105  2 105  2 105  2 107  2 109  7 106  5

Data are means  standard deviation. *Significantly different from T0 within the same treatment (p < 0.05). §Significantly different from
other treatments at the same time point (p < 0.05).

© 2016 Association of Veterinary Anaesthetists and the American College of Veterinary Anesthesia and Analgesia 9
Dexmedetomidine–opioids in sheep LPB Borges et al.
2009). The opioid dose rates chosen were the same
as used in a previous study in sheep (de Carvalho
et al. 2015). Superior sedation was expected in
sheep administered dexmedetomidine with an opioid
compared with dexmedetomidine alone. However,
although sedation was prolonged, methadone, mor-
phine and butorphanol did not increase the sedation
score. The addition of tramadol to dexmedetomidine
had no impact on the degree or duration of sedation.
This is in contrast to the previous study in which
sedation was enhanced when an opioid was com-
bined with xylazine. The duration for collection of
data was based on the reported duration of sedative
effects of morphine, methadone and tramadol in
combination with dexmedetomidine in dogs (Car-
doso et al. 2014), and the monitoring period of
120 minutes exceeded the duration of sedation of
the drug combinations studied and was sufficient to
effectively compare the treatments.
The central nervous system (CNS) excitatory
effects of opioids administered alone or in combina-
tion with a2-agonists in ruminants have been
described (Waterman et al. 1990, 1991; Levine
et al. 1992; Lin & Riddell 2003; Edmondson et al.
2012; Verbeek et al. 2012; de Carvalho et al.
2015). Lin & Riddell (2003) reported that the IV
injection of butorphanol alone to cattle induced
agitation, vocalization, distress and violent kicking
for 2–3 minutes after injection, and that the addi-
tion of detomidine appeared to suppress this excita-
tory effect. The administration of tramadol IV to
alpacas resulted in severe CNS excitation: hyperes-
thesia, tremors, and ataxia (Edmondson et al.
2012). The behavior of sheep after IV morphine
includes an increase of locomotor activity, vocaliza-
tion and escape behavior (Verbeek et al. 2012).
Signs of CNS excitation were observed in the sheep in
the study presented here following the administra-
tion of opioids, similar to those reported after
administration of xylazine and opioids (de Carvalho
et al. 2015). Not only may excitation influence the
degree of sedation, but the behavior may also mimic
pain-related behavior and obscure pain assessment.
Heart rate in all treatments was significantly
reduced at almost all time points when compared
with baseline. This was expected due to the cardio-
vascular effects of a2-agonists and in agreement with
findings in other species (Murrell & Hellebrekers
2005; Cardoso et al. 2014). Initially after adminis-
tration of dexmedetomidine, hypertension occurs
due to peripheral vasoconstriction, followed by an
increase in vagal tone and a fall in HR. Blockade of
10 © 2016 Association of Veterinary Anaesthetists and the American College of Veterinary Anesthesia and Analgesia
sympathetic outflow from the CNS leads to a longer
period of bradycardia (Murrell & Hellebrekers 2005).
Opioids may potentiate a reduction in HR by
vagomimetic effects (Benyamin et al. 2008). How-
ever, in conscious goats, methadone administration
alone (0.2 mg kg 1 IV or 0.6 mg kg 1 subcuta-
neously) did not reduce HR (Ols
en et al. 2013).
Similarly, butorphanol (0.5 mg kg 1) IV adminis-
tered alone to conscious sheep did not affect HR
(O’Hair et al. 1988). In the present study, when
opioids were combined with dexmedetomidine there
was no significant difference among treatments and
the majority of the fall in HR can be attributed to
dexmedetomidine alone. Hypotension following the
administration of xylazine to sheep has been
reported (Aziz & Carlyle 1978), but was not
measured in other studies (Grant & Upton 2001;
de Carvalho et al. 2015). A delayed decrease in
blood pressure was measured after IV administration
of medetomidine in sheep, but the reduction in MAP
did not appear to be clinically significant (Bryant
et al. 1998). Dexmedetomidine administered intra-
muscularly (IM) to conscious sheep did not signifi-
cantly affect blood pressure (K€astner et al. 2001a),
and hypotension did not occur after IV administra-
tion in the sheep in the present study. The changes
in HR and MAP reported here are similar to the
changes observed after administration of xylazine
and the same opioids (de Carvalho et al. 2015).
The respiratory depressant effects of dexmedeto-
midine have been reported in humans (Belleville et al.
1992) and horses, although this is mild hypercapnia
(Bettschart-Wolfensberger et al. 2005). In humans,
opioids exhibit a dose-dependent effect on the
respiratory system (Gutstein & Akil 2006), but in
animals this is less apparent (Dugdale 2010).
Evidence in ruminants is relatively sparse. Butor-
phanol (0.2 mg kg 1) IV had no effect on blood
gases in healthy sheep, but fentanyl induced a
short duration of respiratory depression (Waterman
et al. 1990, 1991).
In the present study, PaCO2 increased after
administration of dexmedetomidine alone or in
combination with butorphanol or methadone,
although the degree of hypoventilation was mild
and unlikely to be of clinical significance. These
findings are similar to results from sheep adminis-
tered xylazine with methadone or morphine (de
Carvalho et al. 2015). No significant changes in fR
were recorded in sheep after IM administration of
dexmedetomidine (K€ astner et al. 2001a). In con-
trast, methadone administered IV to pygmy goats

resulted in a decrease in PaCO2 and an increase in fR
associated with mild CNS excitatory effects (Neal &
Olsen 1980).
Hypoxemia is often observed in sheep after admin-
istration of dexmedetomidine, even with low doses,
although there may be variation among individuals
(K€astner 2006; K€astner et al. 2007). Several mech-
anisms have been proposed for a2-agonist-induced
hypoxemia in sheep, including intense venous spasm
mediated via adrenoreceptor agonism, pulmonary
congestion and alveolar capillary rupture, culminat-
ing in an inflammatory response (Bacon et al. 1998;
K€astner et al. 2007). In the present study, there were
significant reductions in PaO2, but the magnitude of
the changes differed between animals. Recumbency
following drug administration occured in all treat-
ments except DT, and therefore a positional influence
on gas exchange may have occurred. Lateral recum-
bency induces a fall in arterial oxygenation when
compared with standing sheep (Mitchell & Williams
1977). In the present study, clinically relevant
reductions in PaO2 values were observed in individ-
ual animals, and therefore oxygen supplementation
might be required in some sheep, especially when
working at high altitude, as in this study.
Temporal increases in pH, [HCO3 ] and BE were
observed in the sheep in this study. Significant
increases in pH were longest-lasting in sheep treated
with dexmedetomidine alone. Epidural administra-
tion of xylazine in sheep has been associated with
increases in pH and bicarbonate, indicative of a
metabolic alkalosis; the authors did not speculate as
to why this may have occurred (Aminkov &
Hubenov 1995). Horses administered a 3 hour
infusion of xylazine or romifidine had documented
increases in pH, bicarbonate and BE, probably from a
urinary loss of chloride (Ringer et al. 2013). In our
study there were no significant chloride changes and
we cannot corroborate this hypothesis in sheep and
the cause remains unclear. Increased pH may
explain the rise in PaCO2 observed in some sheep
in this study – if hydrogen ion content falls,
compensation occurs by hypoventilation and an
increase in carbon dioxide attenuating the alkalosis.
However, it is likely that sheep had a mixed acid–
base disturbance with concurrent metabolic alkalo-
sis and respiratory acidosis.
In conclusion, the degree of sedation resulting
from combinations of IV dexmedetomidine
(0.005 mg kg 1) and either butorphanol (0.1 mg
kg 1), methadone (0.5 mg kg 1), morphine
(0.5 mg kg 1) or tramadol (5.0 mg kg 1) was sim-
© 2016 Association of Veterinary Anaesthetists and the American College of Veterinary Anesthesia and Analgesia
Dexmedetomidine–opioids in sheep LPB Borges et al.
ilar to that from the administration of dexmedeto-
midine alone. Changes in HR, MAP, fR, PaCO2 and
BE were not clinically significant. However,
decreases in PaO2 and SaO2 were measured at
15 minutes after treatment administration, which
suggests that oxygenation or pulse oximetry should
be monitored and oxygen supplementation provided
if necessary. As the number of animals and drugs
doses used in this study were limited, further
investigations of different dose rates may identify a
more effective combination for clinical use.
Acknowledgements
The authors would like to thank the Fundacßa ~o de
Amparo a Pesquisa do Estado de S~ao Paulo (FAPESP;
2012/23664-9) for financial support, Ana M
arcia
Zago and the Design Department of the University of
Franca for drafting the figure and Professor Daniel
Kan Honsho for granting the experimental area.
Authors’ contributions
LPBB, LTN, LLC, SAC: conception and development
of the study design, data collection; AA, EM-J:
analysis and interpretation of the data; SAC, AA,
EM-J: drafting the manuscript; AA, EM-J: critical
revisions of the manuscript.
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Received 29 July 2015; accepted 18 November 2015.