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). References Banks WJ (1993) Applied veterinary histology, 3rd edn. 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