Comp Clin Pathol (2008) 17:197–201
DOI 10.1007/s00580-008-0720-8
ORIGINAL ARTICLE
Haematology and serum biochemistry of golden eagle
(Aquila chrysaetos) in Iran
S. Nazifi & A. Nabinejad & M. Sepehrimanesh &
S. L. Poorbaghi & F. Farshneshani & M. Rahsepar
Received: 28 October 2007 / Accepted: 28 January 2008 / Published online: 5 March 2008
# Springer-Verlag London Limited 2008
Abstract Haematological and serum biochemical values
were estimated in blood samples collected from 21
apparently adult golden eagles (Aquila chrysaetos) of both
sexes. The mean values of red blood cells, packed cell
volume, haemoglobin, white blood cells, heterophils,
lymphocytes, monocytes and eosinophils were 1.63 ±
0.11×1012/l, 0.47±0.009 l/l, 91.73±1.52 g/l, 24.31±1.97×
109/l, 4.40±0.22×109/l, 16.81±0.65×109/l, 0.99±0.19×
109/l and 2.10±0.30×109/l, respectively. The leucocytes
had 69.14%, 4.09%, 18.12% and 8.65% lymphocytes,
monocytes, heterophils and eosinophils, respectively. The
results of serum biochemistry in the golden eagle indicated
that the concentrations of glucose, total protein, albumin,
total globulin, cholesterol, triglyceride, uric acid, blood urea
nitrogen, creatinine, calcium, phosphorous, aspartate aminotransferase, alanine aminotransferase, creatine kinase,
lactate dehydrogenase and alkaline phosphatase were
16.42 ± 0.73 mmol/l, 49.76 ± 1.35 g/l, 20.46 ± 0.79 g/l,
29.30±1.47 g/l, 2.14±0.09 mmol/l, 2.04±0.08 mmol/l,
457.67 ± 97.46 μmol/l, 2.74 ± 0.17 mmol/l, 53.27 ±
3.87 μmol/l, 2.37 ± 0.24 mmol/l, 1.73 ± 0.08 mmol/l,
293.24±18.96 IU/l, 28.21±2.36 IU/l, 411.29±58.37 IU/l,
1,209.89±21.73 IU/l and 67.31±5.29 IU/l, respectively.
There were no significant differences between haematological
S. Nazifi (*) : M. Sepehrimanesh : S. L. Poorbaghi :
F. Farshneshani : M. Rahsepar
Department of Clinical Studies, School of Veterinary Medicine,
Shiraz University,
P.O. Box 1731, Shiraz, 71345, Iran
e-mail: nazifi@shirazu.ac.ir
A. Nabinejad
Department of Avian Researches, Razi Institute, Isfahan Branch,
Isfahan, Iran
and serum biochemical parameters of male and female golden
eagles (P>0.05).
Keywords Haematology . Serum biochemical parameters .
Golden eagle (Aquila chrysaetos)
Introduction
The golden eagle is one of the world's largest living birds.
Traditionally, the health status of wild populations of birds
has been assessed by measurement of population size,
reproductive success or annual survival. These methods of
assessing population health are both practical and valuable,
but enable biologists to recognize problems only after a
large number of individuals are affected. An additional
method for determining animal health which has the
advantage of detecting problems prior to impact on colonies
or populations is blood sampling and analysis (Newman et
al. 1997). Haematological and serum biochemical values
can be obtained easily and are useful in determining the
health or general condition of birds. To rehabilitate
individual raptors or to breed and release endangered birds
of prey, knowledge of their normal concentrations of blood
constituents is of paramount importance. The golden eagle
is a vulnerable species that could become endangered if real
protection is not afforded by the Government Agency for
the Conservation of Nature. Published researches on
haematological and serum biochemical status of domestic
species of birds are numerous (Hodegs 1979; Puls 1988;
Campbell 1995; Nazifi and Vesal 2003; Thrall 2004).
However, more attention has been recently paid to
haematological and biochemical data of wild species of
birds (Hawkey et al. 1983; Sturkie 1986; Puerta et al. 1989;
Peinado et al. 1992; Bounous et al. 2000; Hauptmanova et
198
Comp Clin Pathol (2008) 17:197–201
Table 1 Mean ± SEM of haematological values in golden eagle
(Aquila chrysaetos; n=21)
Parameters
Value
RBC (1012/l)
Hb (g/l)
PCV (l/l)
WBC (109/l)
Heterophil (×109/l)
Lymphocyte (×109/l)
Monocyte (×109/l)
Eosinophil (×109/l)
1.63±0.11
91.73±1.52
0.47±0.009
24.31±1.97
4.40±0.22
16.81±0.65
0.99±0.19
2.10±0.30
RBC red blood cell, Hb haemoglobin, PCV packed cell volume, WBC
white blood cell
al. 2002; Padilla et al. 2003). There is a paucity of
information about the haematological and serum biochemical parameters of the eagle (Polo et al. 1992; Kaneko et al.
1997; Garcia-Montijano et al. 2002). We are aware that
there is no published paper on the haematological and
serum biochemical parameters of the golden eagle. This
paper describes the haematology and serum biochemical
parameters in clinically healthy golden eagles (Aquila
chrysaetos) in Iran.
Materials and methods
Twenty-one clinically healthy adult wild-caught golden
eagles (A. chrysaetos) from both sexes (nine females and 12
males) in Isfahan province of Iran were used in the present
study. Blood samples were collected from the brachial vein
of the birds using 23-gauge needle. For haematological
parameters, blood samples were taken into vacutainers
containing EDTA as an anticoagulant. For the serum
biochemical analysis, blood samples were collected into
vacutainers, and serum was separated by centrifugation at
750×g for 15 min and stored in deep freeze (−20°C) until
use. Total erythrocyte count, total leucocyte count, haemoglobin concentration and haematocrit value were determined using avian haematological techniques (Coles and
Campbell 1986). For differential leucocyte counts, the
blood smears were prepared and stained with Giemsa stain
(Jain 1986). Serum biochemical analysis including total
protein was done by the Biuret method, albumin by the
bromcresol green method, total globulin by the difference
of total protein and albumin, cholesterol by a modified
Abell-Kendall/Levey-Brodie (A-K) method, triglyceride by
the enzymatic procedure of McGowan et al. (1983),
creatinine by the Jaffe method, uric acid by the phosphotungstic acid method, calcium by the orthocresolphthalein
method, inorganic phosphorus by the ammonium molybdate method, glucose by the o-toluidine method, urea
nitrogen by the diacetyl monoxime method, creatine kinase
(CK) by the Sigma colorimetric (modified Hughes) method,
alkaline phosphatase (ALP) by the modified method of
Bowers and McComb, aspartate aminotransferase (AST)
and alanine aminotransferase (ALT) activities by the
colorimetric method of Reitman and Frankel and lactate
dehydrogenase (LDH) by the Sigma colorimetric (Cabaud–
Wroblewski) method. All the enzyme activities were
measured at 37°C and the results have been presented in
units per litre (Burtis and Ashwood 1994). Biochemical
parameters were measured using a standard autoanalyser
with veterinary software (Cobas-Mira, ABX-Diagnostics,
Japan). Data were analysed by independent t test, using
SPSS/PC software. All values were expressed as mean ±
standard error (SEM), and P<0.05 was determined as
statistically significant.
Results
The mean ± SEM of haematological and serum biochemical
values in golden eagles (A. chrysaetos) are presented in
Tables 1 and 2, respectively. There were no significant
differences between haematological and serum biochemical
parameters of male and female golden eagles (P>0.05).
The morphology of the cellular elements in the blood of
golden eagles was similar to those in other birds. Golden
eagle heterophils were large spherical cells with two or
three segments in the nucleus. Their cytoplasm contained
pinkish specific granules (Fig. 1). Eosinophils were also
spherical cells containing nuclei with two or three seg-
Table 2 Mean ± SEM of serum biochemical values in golden eagle
(Aquila chrysaetos; n=21)
Parameters
Value
Glucose (mmol/l)
AST (IU/l)
CK (IU/l)
ALP (IU/l)
Albumin (g/l)
Cholesterol (mmol/l)
Creatinine (mmol/l)
Calcium (mmol/l)
Uric acid (mmol/l)
ALT (IU/l)
LDH (IU/l)
Total protein (g/l)
Total globulin (g/l)
Triglyceride (mmol/l)
BUN (mmol/l)
Phosphorous (mmol/l)
16.42±0.73
293.24±18.96
411.29±58.37
67.31±5.29
20.46±0.79
2.14±0.09
53.27±3.87
2.37±0.24
457.67±97.46
28.21±2.36
1,209.89±21.73
49.76±1.35
29.30±1.47
2.04±0.08
2.74±0.17
1.73±0.08
AST aspartate aminotransferase, CK creatine kinase, ALP alkaline
phosphatase, ALT alanine aminotransferase, LDH lactate dehydrogenase, BUN blood urea nitrogen
Comp Clin Pathol (2008) 17:197–201
Fig. 1 Heterophils in the blood smear of golden eagle (Giemsa, ×930)
ments, and their cytoplasm was occupied by roundedspecific acidophilic granules. Basophils were spherical cells
with an eccentric round or oval-shaped nucleus. Their
cytoplasm contained highly basophilic specific granules.
The lymphocytes in golden eagle were spherical cells
which contained a large rounded nucleus with dense
chromatin. The nuclei may show a small depression and
fills nearly the entire cell, leaving only a narrow rim of
cytoplasm. They were classified as medium or small. The
monocytes were large spherical cells with an ovoid or beanshaped nucleus, located eccentrically (Fig. 2).
Discussion
Blood analysis in combination with other diagnostic
methods enables a veterinarian to assess the health of
individuals and identify organs affected by infectious
diseases, toxins and metabolic or nutritional diseases
(Newman et al. 1997). Data presented in this study were
obtained from free-living golden eagles in Isfahan province
of Iran. Due to the precarious status of this bird, raptor
recuperation centres and zoological institutions are trying to
save them from extinction. A haematological and clinical
chemical “screening” is recommended in veterinary diagnosis for all captive-breeding projects and, when possible,
for the populations of vulnerable species (Polo et al. 1992).
The findings of this study revealed that the shape and
characteristic features of erythrocytes and leucocytes in
golden eagles were similar to those in other birds,
especially chickens (Mukhergee et al. 1971; Maxwell and
Siller 1972; Maxwell 1973, 1974; Hodegs 1974, 1977;
King and McLelland 1984; Campbell 1988; Banks 1993;
Tadjalli et al. 1997; Garcia-Montijano et al. 2002).
In this study, haematocrit value in golden eagle was 0.47±
0.009 l/l which was similar to those found in carnivorous
birds (Balasch et al. 1974; Hawkey et al. 1983; Feldman et
al. 2000; Garcia-Montijano et al. 2002; Lanzarot et al.
199
2005). The mean value of total red blood cell count in
golden eagle was lower than the values reported in
carnivorous birds (Balasch et al. 1974; Hawkey et al.
1983; Feldman et al. 2000; Lanzarot et al. 2005; Mostaghni
et al. 2005). The haemoglobin value of golden eagle was
91.73± 1.52 g/l which was lower than that found in
carnivorous birds (Balasch et al. 1974; Hawkey et al.
1983; Feldman et al. 2000; Lanzarot et al. 2005; Mostaghni
et al. 2005). The total number of leucocytes in golden eagle
was 24.31±1.97 which was higher than those in carnivorous birds (Balasch et al. 1974; Hawkey et al. 1983;
Feldman et al. 2000; Lanzarot et al. 2005). In the golden
eagle, lymphocytes were the most abundant leucocyte
representing 69.14±2.67% of the leucocyte differential
count. The heterophil was the next most frequently
observed leucocyte found in the blood of golden eagle
representing an average of 18.12% of leucocytes (Table 1).
The percentage of heterophil in the golden eagle was
approximately lesser than those found in carnivorous birds
(Balasch et al. 1974; Hawkey et al. 1983; Feldman et al.
2000; Lanzarot et al. 2005). The percentages of lymphocytes, monocytes and eosinophils were approximately
similar to those found in carnivorous birds (Balasch et al.
1974; Hawkey et al. 1983; Feldman et al. 2000; GarciaMontijano et al. 2002; Lanzarot et al. 2005).
The level of total protein in birds is less than that of
mammalians with a normal range from 3 to 5 g/dl for most
birds which with the results of the present study is in
agreement (Coles and Campbell 1986; Coleman et al. 1988;
Kaneko et al. 1997; Khazraiinia et al. 2006). The concentration of serum total protein in golden eagle was in the upper
limit of the normal range of the Spanish imperial eagle and
hawk (Kaneko et al. 1997; Garcia-Montijano et al. 2002).
The following serum protein fractions were identified:
albumin and total globulin. These are the same as those
detected in four species of birds of prey (Polo et al. 1992).
The concentration of serum albumin and globulin in golden
eagle was similar to the normal value of the Spanish imperial
Fig. 2 Monocyte in the blood smear of golden eagle (Giemsa, ×930)
200
eagle (Garcia-Montijano et al. 2002) and most birds (Puerta
et al. 1989; Kaneko et al. 1997; Thrall 2004; Lanzarot et al.
2005; Mostaghni et al. 2005). Glucose concentration in
healthy birds is generally higher than that of mammalians at
around 200–500 mg/dl. This may be explained by the fact
that whereas insulin plays a key role in mammalian glucose
homoeostasis, glucagon plays the major role in the maintenance of normal avian blood glucose concentration. It is
known that wild birds tend to have lower blood glucose than
captive birds (Lewandowski et al. 1986). In our study,
glucose concentration was lower than that in most birds but
it is difficult to know if these variations are characteristic of
individual species or a consequence of a diet poor in glucose.
The concentration of serum glucose in the golden eagle was
similar to the normal value of the Spanish imperial eagle and
hawk (Kaneko et al. 1997; Garcia-Montijano et al. 2002).
Uric acid is the major end product of nitrogen metabolism in
birds which results in significantly higher uric acid levels in
birds than in mammals. In the present study, uric acid
concentrations were in the normal ranges of the Spanish
imperial eagle and hawk (Kaneko et al. 1997; GarciaMontijano et al. 2002) which is influenced by species, age
and diet (Thrall 2004). In our study, sex had no significant
effect on serum concentration of uric acid. Alanine aminotransferase and aspartate aminotransferase activities have a
wide range in many species of birds. Our results were in
reference range of most birds (Kaneko et al. 1997; Thrall
2004). It has been reported that there are some variations in
the cholesterol level among various avian species (Coles and
Campbell 1986; Coleman et al. 1988; Kaneko et al. 1997;
Khazraiinia et al. 2006). The mean concentration of
cholesterol in golden eagle was lower than that in most
birds (Coles and Campbell 1986; Coleman et al. 1988; Thrall
2004; Khazraiinia et al. 2006). There was no reference range
for the cholesterol concentration of eagle in the literature
review. It is known that serum cholesterol increases in birds
fed with a diet rich in fat and poor in protein; but it is
difficult to know if these variations are characteristic of
individual species or a consequence of a diet rich in fat.
There was no reference range for serum triglyceride of birds
including eagle in the literature review. Serum creatinine
levels in golden eagle were similar to previously reported
values in other birds (Coles and Campbell 1986; GarciaRodriguez et al. 1987; Coleman et al. 1988; Thrall 2004).
The concentration of serum creatinine in our study was lower
than the eagle but similar to hawk (Kaneko et al. 1997).
Serum calcium concentration in golden eagle was similar to
the normal values reported for eagle and hawk (Kaneko et al.
1997). There was no reference range for CK activity of eagle
in the literature review. The activity of CK in golden eagle
was higher than that in other birds (Coles and Campbell
1986; Garcia-Rodriguez et al. 1987; Coleman et al. 1988;
Thrall 2004). A variety of insults (pathologic and iatrogenic)
Comp Clin Pathol (2008) 17:197–201
may damage muscle fibers and cause the release of CK from
muscle fibers. Injury to organs and tissues containing smooth
muscle may cause increased serum CK activity but to a
lesser extent than with striated muscle damage (Stockham
and Scott 2002), but it is difficult to understand that the
increase was due to muscle damage or other factors. There
was no reference range for ALP activity of eagle in the
literature review. Serum alkaline phosphatase of the golden
eagle was higher than that reported for other species of birds
(Coles and Campbell 1986; Garcia-Rodriguez et al. 1987;
Coleman et al. 1988; Thrall 2004). Serum ALP activity in
birds primarily results from osteoblastic activity. Therefore,
ALP activity is not useful in the detection of hepatobiliary
disease (Thrall 2004). Serum phosphorous concentration of
the golden eagle was in normal range of most birds (Coles
and Campbell 1986; Garcia-Rodriguez et al. 1987; Coleman
et al. 1988; Kaneko et al. 1997; Thrall 2004) which may be
dependent on age (Stockham and Scott 2002). In the present
study, serum LDH activity was similar to the Spanish
imperial eagle (Garcia-Montijano et al. 2002) but higher
than the hawk (Kaneko et al. 1997). Serum LDH activity is
influenced by species (Thrall 2004). Urea generally is
considered to have limited diagnostic value in the detection
of renal disease in birds compared to uric acid. Blood urea
concentration of normal noncarnivorous birds ranges between 0 and 10 mg/dl (Kaneko et al. 1997; Thrall 2004).
Plasma urea concentration increases in birds after ingestion
of high protein (Thrall 2004). There was no reference range
for blood urea nitrogen of eagle in the literature review.
Serum urea nitrogen in golden eagle was higher than that in
other birds (Coles and Campbell 1986; Garcia-Rodriguez et
al. 1987; Coleman et al. 1988; Kaneko et al. 1997; Thrall
2004; Khazraiinia et al. 2006).
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