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Effects of xylazine–ketamine–diazepam
anesthesia on blood cell counts and plasma
biochemical values in sheep and goats

ARTICLE in COMPARATIVE CLINICAL PATHOLOGY · JANUARY 2010

DOI: 10.1007/s00580-009-0923-7

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Zuhair Bani Ismail Khaleel Jawasreh

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Comp Clin Pathol (2010) 19:571–574
DOI 10.1007/s00580-009-0923-7
ORIGINAL ARTICLE
Effects of xylazine–ketamine–diazepam anesthesia on blood
cell counts and plasma biochemical values in sheep and goats
Zuhair Bani Ismail & Khaleel Jawasreh &
Ahmad Al-Majali
Received: 4 August 2009 / Accepted: 8 October 2009 / Published online: 21 October 2009
# Springer-Verlag London Limited 2009
Abstract To examine the effects of xylazine, ketamine,
diazepam anesthesia on clinical pathology parameters in
sheep and goats, seven adult, healthy, non-pregnant Awassi
sheep weighing 40–60 kg and seven adult, healthy non-
pregnant Damascus breed goats weighing 35 to 55 kg were
used. Anesthesia was induced using 0.1 mg/kg xylazine,
5 mg/kg ketamine, and 0.25 mg/kg diazepam as a single
intravenous injection. Blood cell counts and plasma biochem-
ical analysis were performed before anesthesia (T0), 2 h after
recovery, 24 h after recovery, and 5 days later. Hematological
analysis indicated that there was a significant increase in
neutrophil percentages (P≤0.05) in sheep and goats at 2 h
and 24 h after recovery. There was a mild lymphopenia and a
significant leukocytosis at 24 h after recovery in both species.
Packed cell volume was significantly increased at 24 h and at
2 h and 24 h after recovery in sheep and goats, respectively,
while total red blood cell count was significantly decreased at
24 h after recovery in sheep and goats. There were no
significant changes in hemoglobin, blood urea nitrogen,
creatinine, total protein, and albumin concentrations. Glucose
was significantly elevated at 2 h after recovery in goats only.
Aspartate aminotransferase, alkaline phosphatase, and
gamma-glutamyl transpeptidase were not significantly
changed at any sampling point. It is therefore concluded that
Z. Bani Ismail (*) : A. Al-Majali
Department of Veterinary Clinical Sciences, Faculty of Veterinary
Medicine, Jordan University of Science and Technology,
Irbid 22110, Jordan
e-mail: zuhair72@just.edu.jo
K. Jawasreh
Livestock and Range Land Research,
National Center for Agriculture Research and Extension,
Amman, Jordan
a combination of xylazine, ketamine, and diazepam can be
used to induce short term anesthesia in sheep and goats with
minimum effects on clinical laboratory parameters.
Keywords Injectable anesthesia . Xylazine . Ketamine .
Diazepam . Sheep . Goats . Clinical pathology
Introduction
Xylazine (alpha2 adrenergic agonists) is the most commonly
used anesthetic agent in ruminants. It usually induces a dose-
dependent sedative-hypnotic effect, and it can be combined
with other anesthetic agents to obtain surgical anesthesia.
The use of xylazine in ruminants is associated with moderate
to severe cardiopulmonary depression, prolonged induction
and recovery, ineffective in excited animals, and at higher
doses, rumen stasis, apnea and bradycardia (Ewing 1990;
Riebold 1995; Wallincford et al. 1996).
Ketamine is an N-methyl-D-aspartate receptor antagonist
(Ersek 2004). It induces a state referred to as dissociative
anesthesia. Ketamine has a wide range of effects in animals,
including analgesia, anesthesia, elevated blood pressure,
and bronchodilation. Ketamine is primarily used for the
induction and maintenance of general anesthesia, usually in
combination with other sedative drugs such as xylazine or
butorphanol (Lin 1996; Lin and Pugh 2002).
Diazepam is a benzodiazepine derivative drug. It
possesses anxiolytic, anticonvulsant, hypnotic, sedative,
skeletal muscle relaxant, and amnestic properties
(Mandrioli et al. 2008). A combination of diazepam and
ketamine with or without xylazine is frequently used to
induce general anesthesia in small ruminants. Cardiopul-
monary side effects of using various anesthetic reagents
have been studied in animals (Stegmann 1999; Taylor et al.
572 Comp Clin Pathol (2010) 19:571–574

Table 1 Hematological and

plasma biochemical values Parameter Time of sampling
(mean±SD) in Awassi sheep
following xylazine–ketamine– T0 2h 24 h 5 days
diazepam anesthesia (n=7)
RBC (×106) 9±3 8.9±1.5 8.2±2.1a 8.7±2.9
PCV (%) 28±2 28.4±1.3 33±5a 29±2.5
Hb (g/dl) 10±1.5 10.2±2 9.7±1.4 10.4±1.3
WBC (×103) 7±2.5 7.2±2 8.3±3.5a 7.5±2.8
Neutrophils (%) 40±12 55±13.5a 54±11a 42±12.5
Lymphocytes (%) 49±18 43±9 44±12 48±17
BUN (mg/dl) 15±4 14.2±2.8 14.9±3 15.2±1.8
Creatinine (mg/dl) 0.5±0.3 0.7±0.1 0.5±.2 0.5±0.4
Glucose (mg/dl) 68±9 75±12 70±9 72±13
Total protein (g/dl) 6.2±0.5 6.5±.1 6.3±.4 6±0.8
Albumin (g/dl) 2.3±0.5 2±0.5 2.1±0.9 2±0.2
AST (U/ml) 55±10 49±15 53±17 51±12
ALP (U/ml) 120±32 117±12 123±9 125±15
a
Indicate significant changes GGT (U/ml) 31±12 23±10 29±12 33±14
(P≤0.05)

1992; McEwen et al. 2000). However, little information is
available on the systemic effects of using xylazine–
ketamine–diazepam combination to induce and maintain
general anesthesia in small ruminants. Undesirable systemic
effects due to this combination in various organs can be
revealed by studying the changes in biochemical and/or
hematological parameters (Topal et al. 2003; Steffey et al.
1993). Thus, the aim of this study was to examine the
clinical pathology effects of xylazine–ketamine–diazepam
anesthesia in Awassi sheep and Demascus breed goats.
Materials and methods
Experimental design
Seven healthy, adult non-pregnant Awassi sheep weighing
40–60 kg and seven adult, healthy non-pregnant Dam-
ascus breed goats weighing 35 to 55 kg were used.
Animals were kept at the National Center for Agriculture
Research and Extension farm at Al-Khanasri region in
North Jordan. The study was conducted during April of

Table 2 Hematological and

plasma biochemical values Parameter Time of sampling
(mean±SD) in Damascus goat
following xylazine–ketamine– T0 2h 24 h 5 days
diazepam anesthesia (n=7)
RBC (×106) 11.3±2.5 10.5±3 9.3±2.2a 10.8±1.2
a
PCV (%) 27±2 29.7±1.6 28.5±2.7a 27±0.6
Hb (g/dl) 9±1.3 8.9±1.4 9.2±1.6 9±1.8
WBC (×103) 8±4 8.3±3.5 9.9±3a 8.2±2.9
Neutrophils (%) 37±9 49±12a 47±9a 39±7
Lymphocytes (%) 62±13 59±12 58±9.5 60±15
BUN (mg/dl) 25±6 24±3 25.5±5.6 23±6.5
Creatinine (mg/dl) 0.7±0.8 0.76±0.7 0.6±0.9 0.67±0.8
Glucose (mg/dl) 72±16 78±14a 69±23 71±13
Total protein (g/dl) 7.3±0.6 7.5±0.1 7.4±0.9 7.2±0.5
Albumin (g/dl) 3.0±0.7 2.9±0.3 3.0±0.4 2.8±0.8
AST (U/ml) 41±22 36±28 44±23 49±18
ALP (U/ml) 55±17 53±14 58±20 51±15
a
Indicate significant changes GGT (U/ml) 33±12 29±16 31±11 33±11
(P≤0.05)
Comp Clin Pathol (2010) 19:571–574
2008. Animals were fed an adequate diet of hay and grain
twice daily, in addition to a limited daily access to
pasture. Animals were routinely administered anthelmen-
tic prophylaxis four times per year. Animals were
thoroughly examined for abnormalities in different body
systems. Animals with abnormalities were excluded. In
addition, whole blood was collected for hematological
and biochemical analysis before the start of the study.
Only animals that showed normal blood parameters were
included in the study.
Before induction of anesthesia, animals were kept off
feed and water for 12 h and 2 h, respectively. Anesthesia
was induced using 0.1 mg/kg, 5 mg/kg, and 0.25 mg/kg
xylazine, ketamine, and diazepam, respectively, as a single
intravenous injection in the same syringe. Whole blood for
hematological and plasma biochemical analyses was with-
drawn from all animals from the jugular vein. Blood cell
counts and plasma biochemical analysis were performed
before anesthesia (T0) and at 2 h after recovery, 24 h after
recovery, and 5 days later. Hematological analysis was
performed using electronic cell counter (ABC Vet hematol-
ogy analyzer, ABX Diagnostics, France). Plasma was
collected by centrifugation of blood tubes at 5,000×g for
10 min at 4°C, and stored at −20°C until analyzed. Plasma
was analyzed for TP (Biuret method), albumin (BCG
method), blood urea nitrogen (Colorimetric method), creat-
inine (Kinetic method), glucose (GOD-PAP method), AST
(IFCC method), GGT (Kinetic method) and ALP (Kinetic
method) using commercially available kits and reagents
according to manufacturers' recommendations.
Analysis of data
Data are presented as mean±SD of means. Non-paired
Student's t test was used to examine the significance of
differences between parameters in sheep and goats. The
difference was considered statistically significant at P≤0.05.
Results and discussion
The average surgical anesthesia induced using a combina-
tion of xylazine, ketamine, and diazepam was 67 and
60 min in sheep and goats, respectively. Smooth induction
within approximately 3–5 min was achieved in both
species. Prolonged but smooth recovery however was
observed. In small ruminants, rapid smooth induction and
gradual recovery while producing sufficient depth of
anesthesia have been reported using this combination
(Carrol et al. 1997; Lin and Pugh 2002; Ceylan et al. 2007).
Changes in hematological and biochemical values
following xylazine-ketamine-diazepam anesthesia in
Awassi sheep and Damascus goats are summarized in
573
Tables 1 and 2, respectively. In the hematological analysis,
there were a significant (P≤0.05) increase in total white
blood cell count, neutrophil percentage, and PCV value at
24 h after recovery in sheep. Neutrophil percentage was
also significantly (P≤0.05) elevated at 2 h. In goats, total
white blood cell count, neutrophil percentage, and PCV
value were significantly (P≤0.05) elevated at 24 h after
recovery. In addition, neutrophil percentage and PCV value
were significantly (P≤0.05) elevated at 2 h after recovery.
In both species, there was a non-significant decrease in
lymphocyte percentage at 2 and 24 h after recovery. The
reported increase in total white blood cells and percentage
of neutrophils at 2 and 24 h after recovery is typical of
corticosteroid induced stress response (Carrol et al. 1997).
The increased in PCV value could be due to mild
dehydration.
In the plasma biochemical analysis, blood urea nitrogen,
and creatinine were not significantly elevated at any time
point. In goats, glucose value was significantly elevated at
2 h after recovery and not significantly elevated in sheep.
Mild hyperglycemia has been related to an increase in
adrenaline and/or corticosteroids secretion (Carrol et al.
1997). Total protein values were not significantly elevated
at 2 and 24 h after recovery which may also be explained
by a mild but not significant hemoconcentration.
Liver enzymes including AST, ALP and GGT were not
significantly affected in sheep and goats following xyla-
zine–ketamine–diazepam anesthesia. Relative increase in
AST, ALP and GGT has been reported in sheep, calves, and
horses following prolonged halothane anesthesia due to the
induction of hepatic microsomal enzymes by halothane
(Steffey et al. 1993; Rajaian et al. 2008). It seems that none
of the used drugs in this study induces such hepatic
microsomal changes.
Results of this study show that xylazine–ketamine–
diazepam anesthesia can be used safely in sheep and goats
after a single intravenous injection. However, studies on the
effect of repeated injections of this combination are needed
in the future.
Acknowledgements This project was partially sponsored by the
Deanship of Research at Jordan University of Science and Technol-
ogy. The authors would like to thank the National Center for
Agriculture Research and Extension, for providing the animals used
in this study and for their technical help.
References
Carrol GL, Hartsfield SM, Hambleton R (1997) Anesthetic effects of
tiletamine-zolazepam, alone or in combination with butorphanol,
in goats. JAVMA 211:593–597
Ceylan C, Aydilek N, Ipek H (2007) Effects of tiletamine-zolazepam
anesthesia on plasma antioxidative status and some hematolog-
ical parameters. Acta Vet Hung 55:191–197
574
Ersek R (2004) Dissociative anesthesia for safety's sake: ketamine and
diazepam-a 35-year personal experience. Plast Reconstr Surg
113:1955–1959
Ewing KK (1990) Anesthesia techniques in sheep and goats. Vet Clin
North Am Food Anim Pract 6:759–778
Lin HC (1996) Lumb and Jones' veterinary anesthesia. Williams and
Wilkins, Media, Pennsylvania
Lin HC, Pugh DG (2002) Sheep and goat medicine. Saunders, Pennsylvania
Mandrioli R, Mercolini L, Raggi MA (2008) Benzodiazepine
metabolism: an analytical perspective. Curr Drug Metab
9:827–844
McEwen MM, Gleed RD, Ludders JW, Stokol T, Del Piero F, Erb HN
(2000) Hepatic effects of halothane and isoflurane anesthesia in
goats. JAVMA 217:1697–1700
Rajaian H, Nazifi S, Nayeri K, Saberi FA (2008) Effects of 2 h anesthesia
with halothane on blood cell counts and serum biochemical values
in sheep and calves. Comp Clin Pathol 17:13–16
Comp Clin Pathol (2010) 19:571–574
Riebold TW (1995) Large animal anesthesia: principles and techni-
ques. Iowa State University Press, Ames, Iowa
Steffey EP, Giri SN, Dunlop CL, Cullen LK, Hodgson DS, Willits N
(1993) Biochemical and hematological changes following pro-
longed halothane anesthesia in horses. Res Vet Sci 55:338–345
Stegmann GF (1999) Observations on some cardiopulmonary effects
of midazolam, xylazine and a midazolam/ketamine combination
in the goat. JSAVA 70:122–126
Taylor JH, Botha CJ, Swan GE, Műlders MSG, Grobler MJ (1992)
Tiletamine hydrochloride in combination with zolazepam hydro-
chloride as an anaesthetic agent in sheep. JSAVA 63P:63–65
Topal A, Gul N, Llcol Y, Gorgul OS (2003) Hepatic effects of
halothane, isoflurane or sevoflurane anaesthesia in dogs. JVMA
50:530–533
Wallincford BD, Lancia RA, Soutiere EC (1996) Antagonism of
xylazine in white-tailed deer with intramuscular injection of
yohambine. JWD 32:399–402