2004

Asiatic Herpetological Research

Vol.10, pp. 230-234

An Investigation on the Blood Cells of the Leopard Gecko,
Eublepharis angramainyu (Reptilia: Sauria: Eublepharidae)
MURAT TOSUNOĞLU1, DINÇER AYAZ2, CEMAL VAROL TOK1, AND BA şARAN DÜLGER1
1Çanakkale Onsekiz Mart University, Faculty of Science-Literature, Department of Biology, Terzio lu Campus,ğÇanakkale, Turkey
2Ege University, Faculty of Science, Department of Biology, 35100 Bornova, zmir ,Turkey

Abstract. - In this study, blood cell counts and sizes in three adult Eublepharis angramainyu specimens (one male,two female) collected from SE Anatolia (Sanlurfa – Birecik). The number of erythrocytes in 1 mm3 ranged between
870,000 and 950,000 (average 910,000). The mean total length of erythrocytes was calculated as 20.35 µm, the width
as 10.59, the size as 169.68 µm2; the mean nucleus length as 7.50 µm, the width as 4.15 and the size as 24.47 µm2.
Small lymphocytes had a mean diameter of 9.78 µm, big lymphocytes 13.71 µm, monocytes 15.37 µm, neutrophiles
16.56 µm, eosinophiles 17.59 µm, and basophiles 12.87 µm. The mean length of thrombocytes was measured at 8.82
µm, and the width at 5.93 µm.
Key words. - Eublepharis angramainyu, Sauria, blood cell count, blood smears, erythrocytes, leucocytes, thrombocytes.

Introduction
Eublepharis angramainyu Anderson and Leviton, 1966,
also known as the “leopard gecko”, was first foundbetween Masjid Soleyman and Batsvand in the Khuzesdan province of Iran. The species was reported to range
in the western foothills of the Zagros Mountains andMesopotamia in Iraq and Iran, and NE Syria with a vertical distribution of 300 to 1000 meters (Anderson andLeviton, 1966; Leviton et al., 1992; Disi and Böhme,1996; Anderson, 1999). Studies conducted in recentyears (Göçmen et al., 2002) established that the species
also inhabited SE Anatolia (Sanl1urfa–Birecik).
Although there are a number of studies on the distribution, morphology and ecology of the species (Andersonand Leviton, 1966; Leviton et al., 1992; Disi andBöhme, 1996; Anderson, 1999; Göçmen et al., 2002), aliterature review has not revealed any detailed haematological studies.
Most studies on the haematology of different species are related to blood cell counts (Alder and Huber,1923; Hutchison and Szarski, 1965; Duguy, 1970;
Ar1kan, 1989) and blood cell sizes (Szarski and Czopek,1966; Hartman and Lessler, 1964; Atatür et al., 1998,1999). The number haematological studies related toamphibian and reptile species living in Anatolia has
been increasing in recent years (Arkan, 1989; Atatür etal., 1998, 1999, 2001; Sevinç et al., 2000). In this study,
the number and sizes of blood cells of Eublepharis© 2004 by Asiatic Herpetological Researchangramainyu were determined and photographs of theirblood cells presented.

Materials and Methods
Three adult specimens (one male, two female) examinedin this study were collected near Çiçekalan Village
between 2200-2400 hours at an altitude of 400 m during
the species breeding season (01 July, 2002). Blood samples were taken within the first three days after the specimens were collected live in the wild and brought to thelaboratory.
Blood cell counts were carried out by means of
Neubauer hemocytometer. Hayem solution was used todilute the erythrocytes. Wright-Stained blood smears
were made use of in the measurement (erythrocytes, leucocytes and thrombocytes) and computation of bloodcells. The necessary blood samples were obtained bycardiac (ventriculus) puncture, via heparinized hematocrit capillaries. Blood cell measurements were taken bymeans of a MOB-1-15x ocular micrometer. On eachblood smear, measurements related to 40 randomly chosen erythrocytes (total erythrocyte length, total erythrocyte width, nucleus length and nucleus width) were
made, and the nucleus size was calculated) according to
the formula EL.EW./4 and the nucleus size according to
the formula NL.NW./4 (Duguy, 1970; Atatür et al.,2001). Moreover, micrometric measurements weremade on leucoytes and thrombocytes. Photographs of

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Asiatic Herpetological ResearchA

BCDEFG

HFigure 1: The blood cells of Eublepharis angramainyu. Aerythrocytes, B- small lymphocyte, C- large lymphocyte,
D- monocyte, E- neutrophil, F- eosinophil, G- basophil,H- a cluster of thrombocytes.

blood cells were taken using a Carl Zeiss Jena microscope at 40x magnification.

Results
Male and female specimens were assessed together asthere were no significant differences between them withrespect to the number and size of blood cells. As in otherlizrds, the erythrocytes belonging to Eublepharis angramainyu are also ellipsoidal cells with nuclei. The nuclei
are also ellipsoidal, somewhat regular and centrally

2004

located (Figure 1A). The mean total length of the erythrocytes (L) was calculated as 20.35 µm the width (W) as
10.59 µm, the size (S) as 169.68 µm2; the mean nucleus
length (NL) as 7.50 µm, the width (NW) as 4.15 µm,and the size (NL) as 24.47 µm2. The number of erythrocytes in 1 mm3 of blood ranged between 870,000 and
950,000 (average 910,000) (Table 1).
Lymphocytes have a spherical shape. Both small
and large lymphocytes were examined in the bloodsmears prepared (Figure 1B, C). Large lymphocytes
were 13.71 µm in diameter and had a large cytoplasmiczone and a centrally-located, large, round nucleus. Thecytoplasm is stained pale blue and the nucleus purplish
blue using the Wright Stain. No granule formation wasobserved in the cytoplasm. Small lymphocytes had amean diameter of 9.78 µm (Table 1). The large nucleus
covers the majority of the cell’s area; the cytoplasm is in
the shape of a thin ring. Lymphocytes are the most commonly seen leucocytes in the preparates.
Although resembling the large lymphocytes in size,
monocytes are easily distinguished from the shape ofthe nucleus (Figure 1D). They have a mean diameter of
15.37 µm Granule formation was observed in the cytoplasm (Table 1). The nucleus is not oval, but depressedon one side and occupies at least half of the cell. Thecytoplasm is stained light purple and the nucleus dark
blue. They are the second most common leucocytes
after lymphocytes and neutrophiles.
Neutrophiles are spherical cells with a mean diameter of 16.56 µm (Table 1). Using the Wright Stain, thecytoplasm is stained light blue and the nucleus dark
blue. There are very fine granules in the cytoplasm (Figure 1E). The nucleus is a structure with lobes and segments. They are the most common leucocytes second tolymphocytes.
Eosinophiles have a diameter of 17.59 µm (Table1).
The cytoplasm is stained light blue and the nucleus darkblue. Large, round, bright red granules within the cytoplasm strike eye as the most distinctive characteristic ofthese cells (Figure 1F). The nucleus was seen to have
two lobes. These cells take the fourth place in the preparation of smears after lymphocytes, monocytes and neutrophiles.
Basophiles are oval-shaped with a mean diameterof 12.87 µm (Figure 1G and Table 1). When stained bymeans of the Wright Stain, dark bluish purple granules
within the light blue cytoplasm are in a position marking
the dark blue nucleus. These cells are rarely seen in thepreparates.
Thrombocytes are spindle-shaped cells with a mean
length of 8.82 µm, a width of 5.93 µm (Figure 1H andTable 1). In the Wright-Stained preparates, dark stained
cells with large oval nuclei and small irregular cytoplasic zones form groups of two or more.

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Asiatic Herpetological Research

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Table 1: The established counts, measurements and sizes concerning the blood cells of Eublepharis angramainyu (in m and m2). N: Number of specimens; n: Number of measurements/computings in each specimen; Ext: extreme values; SD and SE: standard deviations and the standard errors of the means,respectively.
Blod Cells

NnExtMeanSDSE

Number of Erythrocytes

23

870,000-950,000

910,000--

Total Erythrocyte Length240

15.00-22.50

20.35

2.15

0.24

Total Erythrocyte Width

240

7.50-12.50

10.59

1.00

0.11

Total Erythrocyte Size

240

91.26-220.78

169.68

26.87

3.00

Nucleus Length240

6.25-8.75

7.50

0.41

0.05

Nucleus Width240

3.25-5.25

4.15

0.52

0.06

Nucleus Size

240

17.86-31.40

24.47

3.44

0.38

Lymphocyte (big) Diameter

220

12.50-16.25

13.71

1.21

0.19

Lymphocyte (small) Diameter

220

7.50-11.25

9.78

1.11

0.17

Monocyte Diameter

240

11.25-20.00

15.37

2.00

0.31

Neutrophile Diameter

240

15.00-20.00

16.56

1.61

0.36

Eosinophile Diameter

220

15.00-20.00

17.59

1.56

0.24

Basophile Diameter25

10.00-15.20

12.87

1.81

0.40

Thrombocyte Length240

7.25-11.25

8.82

1.160.18

Thrombocyte Width240

5.00-8.25

5.93

0.77

0.12

Table 2: The number of erythrocytes in 1 mm3 of blood in different lizard species.
ResearchersSpecies

Number of Erythrocytes

Present study

Eublepharis angramainyu

Duguy (1970)

Hemidactylus turcicus

866,000

Chalcides ocellatus

806,000

Agama atra

1,250,000

Lacerta agilis

945,000-1.420.000

Lacerta viridis

840,000-1,600,000

Anguis fragilis

466,000-1,615,000

870,000-950,000

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Asiatic Herpetological Research2004

Table 3: Sizes of erythrocytes and nuclei in different lizard species according to various researchers (EL/EW: Erythrocyte Length/Erythrocyte Width Ratio, ES: Erythrocyte Size (µm2), NL/NW: Nucleus Length/
Nucleus Width Ratio, NS: Nucleus Size (µm2), N/C: Nuclear surface/Cell surface ratio).
Researchers

Species

EL/EW

ES

NL/NW

NS

N/C

Present study

Eublepharis angramainyu

1.92

169.68

1.80

24.47

0.14

Atatür, et al. (2001)

Ablepharus chernovi

1.87

84.12

2.45

12.01

0.14

Chalcides ocellatus

1.86

91.33

1.98

10.70

0.12

Mabuya aurata

1.90

84.88

2.01

10.02

0.12

Ophiomorus punctatissimus

1.96

92.08

2.30

12.70

0.14

Eumeces schneideri

1.97

92.31

2.81

14.20

0.15

Lacerta rudis

1.63

87.46

1.64

16.66

0.19

Lacerta viridis

1.86

125.00

1.94

14.6

0.11

Eumeces algeriensis

1.61

154.80

2.10

26.4

0.17

Anguis fragilis

1.88

143.90

1.62

22.5

0.14

Sevinç, et al. (2000)
Duguy (1970)

Discussion

Literature Cited

As stated in the ‘Results’ section, E. angramainyu specimens were collected during the mating season and did
not display sexual dimorphism with respect to the number of erythrocytes and sizes. It has been found that
there are significant differences among lizard families
with respect to the number and size of erythrocytes, andthe members of Gekkonidae have the highest number oferythrocytes among lizards (Alder and Huber, 1923;
Hutchison and Szarski, 1965; Duguy, 1970). Values
belonging to the number of blood cells determined indiffeent lizard species (Table 2), by various researchers
were compared with those we obtained for E. angramainyu in the present study, and it was established thatthe number of erythrocytes in 1 mm3 of blood was very
close to that of Hemidactylus turcicus (Gekkonidae)
species, but different from that of other lizard species.
Values we obtained in E. angramainyu with respect
to the sizes of erythrocytes and nuclei were comparedwith those determined for some lizard species (Table 3),and it was found that values concerning the sizes oferythrocytes and nuclei were very close to those of species belonging to Gekkonidae family. When comparedwith the other lizard species, E. angramainyu can besaid to have the largest erythrocytes with respect to thesizes of erythrocytes and nuclei.

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