Asiatic Herpetological Research

2001

Vol. 9, pp. 122-129

The Morphology and Size of Blood Cells of Lacerta rudis bithynica

Murat Sevinc and Ismail H. UGurta§
1Uludagg University, Science and Art Faculty, Department of Biology, 16059 Bursa, Turkey.
correspondence should be addressed e-mail: smurat® uludag.edu.tr

To whom

Abstract.- In this study, the morphology of the blood cells of Lacerta rudis bithynica is described using Wright's

Thesizes of erythrocytes and their nuclei, leukocytes (monocytes, lymphocytes, basophils,andneutrophils,eosinophils), and thrombocytes of L. rudis bithynica were measured using an ocular micrometertechnique.

1600X with an oil immersion objective. The results of this study are compared withworkon
otherprevious
reptile species.
ata magnification ofKey

words.-

Lacertidae,

Lacertarudis

bithynica,

leukocyte,

erythrocyte,thrombocyte,measurement.

morphology.

Introduction
The first studies on the blood of reptiles described thecellular structures, often comparing them with thoseof other vertebrates. Literature on the haematology of
reptilian blood is based on few studies and is usuallyconcernedwithour aim is to describe and measure

In this study,

Europeanspecies(Saintblood cells (erythrocyte, leukocyte, thrombocyte) of
Lacerta rudis bithynica (Squamata: Lacertidae). Thisstudy is the first of its kind on a Turkish species.

Material and Methods

Girons,In1970).

this study,

31

individuals (17 male) of Lacerta

described different circulating blood cells of different

examThe study was carried out between June andAugust 1998. The specimens were collected from
Uludagg (Bursa) at an altitude of 1745 m. Blood wasreptile species (Taylor andKaplan, 1961; Heady andHartman
andLessler, 1964; Szarski
Rogers ,1962;and Czopek, 1966; Duguy, 1970; Saint Girons, 1970,

obtained by cardiac puncture of the lizards (CanfieldCannon et al., 1988, Canfield and Shea, 1996). Other

smears were prepared. Three to five blood smears
were prepared per individual. The smears were air-

Recent studies have concentrated on single species(Tiliqua sp., Cannon et al., 1988; Cyrtopodion sca-brum, Canfield and Shea, 1996). Various authors haveauthors have studied seasonal (Hutton, 1960; Cline

rudis bithynica (Squamata: Lacertidae) wereined.

and Shea, 1988). Immediatelyobtainedinand Waldman, 1962; Haggag and et. al, 1966) or sex-dried and stored untiland Thompson, 1962) variation in thenumber of blood cells of different reptile species. In(Hartmanual (Altlandaddition, authors have studied thenumber of bloodcells of different reptile species (Charipper and Davis,1932; Baker and Cline, 1932; Hutton, 1961; Altlandand Thompson, 1962; Hutchinson and Szarski, 1965;

anddropsofwere allowed to stand for ten minutes at room temperature andwere then washed with distilled water andnuclei, ten thrombocytes,

mals. However, there are no haematological studies
on the Turkish reptiles.

Twelverinsing with a phosphate buffer (pH=6.5). The slides

blood and hematopoiesis of different reptile species

ducted on humans and economically important ani-1964).

Wright's stain were dropped on the slides and allowed
to remain on the slide one and a half minutes before

allowed to dry.

In Turkey, haematological studies have been con-blood wasstained with Wright's stain

Lessler,Engbretson and Hutchinson, 1976). Finally, authors
have studied haemoglobin and hematocrit content of(Altland and Thompson, 1958; Hutton, 1961; Goinand Crawford, 1965; Engbretson and Hutchinson,1976; Newlin and Ballinger, 1976).

after theheparinized capillary tubes, the blood

On each slide fifty mature erythrocytes and theirand ten leukocytes (mono-lymphocyte, eosinophil, basophil and neutrowere measured by means of an ocularphil)micrometer at a magnification of 1600 x with an oilcyte,immersion objective. Cell sizes were calculated fromthe measurements.

Asiatic Herpetological Research

2001

Vol. 9,2014 41
15

•ss

10/""

Figure 1Erythrocytes, an infected erythrocyte and amitosis dvision.
.

ResultsI.

Erythrocytes

Erythrocytes are nucleated, oval cells. Their nuclei are
more or less regular and centrally locatedalso oval,

The cytoplasm of mature erythrocyte1).
appeared light and dark pink and homogeneous under
Wright's stain. The nuclei of mature erythrocytes are(Fig.

chromophilic. In some blood smears, immature erythrocytes are seen. They are characterised by a roundedform, blue cytoplasm and a large nucleus. Mitotic figures are also present and in some smears, intracorpuscular parasites are seen (Fig.

1).

Parasites alter the

shape and size of erythrocytes remarkably.

When

intracorpuscular parasites are seen, immature erythrocytes and mitotic figures are abundant (Fig1).

Intrac-shape and size ofinfected erythrocytes. The shape and size of otherorpuscularparasites

alter

theerythrocytes that are not infected by intracorpuscularparasites are normal.

io15 051123

Vol. 9, p. 124

Asiatic Herpetological Research

2001

Table 2. Erythrocyte dimensions of Lacerta rudis bithynica\oqe\!r\er with the standard deviations in July. EL: Erythrocyte length; EW: Erythrocyte width; ES: Erythrocyte size; NL: Nucleus length; NW: Nucleus width; NS: Nucleussize.

EL ( M m)

EW (pm)

EL/EW

ES (nm2 )NS/ESMaximum

17.08 ±0.799.15 ±0.492.16 ±0.12122.68 ±8.95

0.28 ± 0.02

Minimum

12.81 ±0.79

6.71 ± 0.49

1.53 ±0.1267.47 ± 8.95

0.13 ±0.02

Mean15.05 ±0.79

8.08 ± 0.49

1.86 ±0.12

95.62 ± 8.95

0.18 ±0.02

NL(um)NW (Mm)NL/NW

NS (Mm2Maximum

7.30 ± 0.70

4.27 ± 0.32

2.40 ±0.1724.53 ±2.38

Minimum

5.40 ± 0.70

3.05 ± 0.32

1.33 ±0.17

13.14 ±2.38

Mean6.30 ± 0.70

3.55 ± 0.32

1.79 ±0.1717.67 ±2.38

is stained light red. Eosinophils contain circular toelongate cytoplasmic granules stained brilliant red(Fig 4). Eosinophils are different from neutrophils in
that, eosinophils'

granules are stained bright red andneutrophils' granules were stained dim red.
In June, the mean diameter of eosinophils was
12.82 pm, (±1.71 standard deviations, with a range of

9.93-15.25 um). In July, the mean diameter was
13.29 um (±1.25, 10.98-15.25 um). In August, themean diameter was 13.80 pm (±1.37, 12.20-15.25um).

There were no significant differences in eosinophil
diameters

between females and males. Based on

Tables 4, 5, and 6 and Fig. 8, it appears that there waslittle

monthly variation in the diameter of eosinophils

during the three months.
2. Basophils. Basophils are easily

recognised. They

are small and circular cells. Nuclei stained blue by

Wright technique are commonly obscured by chromophilic circular granules. These cytoplasmic granules are largeand stained dark purple. In the blood<pErythrocytesj^k)2001

Asiatic Herpetological ResearchVol. 9, p. 125

Table 3. Erythrocyte dimensions of Lacerta rudis bithynica together with the standard deviations in August. EL:
Erythrocyte length; EW: Erythrocyte width; ES: Erythrocyte size; NL: Nucleus length; NW: Nucleus width; NS;

Nucleus size.

Vol. 9, p. 126

Asiatic Herpetological Research

2001

Table 6. Leukocyte and thrombocyte measurements of L rudis bithynicamVn the standard derivations in August.

Vol. 9. p. 127

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2001

JuneJuly
E3 Augustlymphocyte

monocyte

neutrohpil

basophil

Leukocytes
Figure 8. Leukocyte diameters of Lacerta rudis bithynica over three months.

Thrombocytes
Ery throe ytes

\Of*-rFigure 9. Erythrocytes and thrombocytes.

86 12ml-\eosinophil

Vol. 9, p. 128

Asiatic Herpetological Research

reported that the neutrophils were not observed, but
the other leukocytes were observed.

Another study on morphological observations on

2001

thrombocytes of blue tongue lizards (Lacertilia: Scincidae, Tiliqua). Anatomia, Histologia, Embryologia.

17:328-342.

the erythrocytes, leukocytes and thrombocytes of blue

Cannon, M. S., D. A. Freed and P. S. Freed. 1996. The

tongue lizards by Canfield and Shea (1988) reported
that all types of leukocytes were observed.

leukocytes of the roughtail gecko Cyrtopodion sca-

Saint Girons (1970) and Canfield and Shea (1988)

divided granulocytes into neutrophils, basophils andeosinophils on the basis of light microscopy. However
Cannon et al. (1996) divided granulocytes into baso-brum: a bright-field and phase-contrast study. Anatomia, Histologia, Embryologia. 25:1 1-14.
Charipper, H. A. and D. Davis. 1932. Studies on thearneth count. A study of the blood cells of Pseudemys

elegans with special reference to the polymorphonu-phils and eosinophils on the basis of bright-field andclear

phase-contrast microscopy.

21:371-382.

leukocytes.

Q.

J.

Experimental

Physiology.

Heady and Rogers (1962) divided leukocytes intoCline, M. J. and T. A. Waldmann. 1962. Effect of tem-neutrophils, small acidophils, eosinophils, lymphocytes and monocytes on the basis of light microscopy

the Society of Experimental Biology and Medicine.

in111:716-718.

perature on red cells in the alligator. Proceedings for

Pseudemys elegans. They, except for monocytes,
gave the sizes of leukocytes and reported that eosinophils and neutrophils were numerous than the otherDuguy, R. 1970. Numbers of blood cells and their

leukocytes. Taylor and Kaplan (1961) also divided

Reptilia, Vol. 3, Morphology C. Academic Press, New

leukocytes into neutrophils, basophils, eosinophils,
lymphocytes and monocytes on the basis of lightYork.

microscopy in turtles.
appears that on the basis of lightmicroscopic findings there are three main types of
granulocytes and two types of agranulocytes in L.
In this study,itrudis bithynica and also the size of all kinds of leukocytes are given in Table 4, 5 and 6.
In the present study, the descriptions

and sizes(Table 4, 5 and 6) of thrombocytes are comparable tootherdescriptions.

CanfieldsandShea(1988)

reported that thrombocyte morphology at the lightmicroscopic level is influenced by the degree ofaggregation and degranulation. Saint Girons (1970)
reported that thrombocytes are small, oval cells characterised by elongate, centrally located highly chro-mophilic nuclei. The cytoplasm is almost colourless(faintly acidophilic) and hence difficult to see in ablood smears. Taylor and Kaplan (1961) reported thesame findings in turtles.

variations. Pp. 93-104. In Gans (ed.),Biology of theVH. Hutchinson. 1976. ErythEngbretson. G. A. androcyte count, hematocrit and haemoglobin content in
the lizard Liolaemus multiformis. Copeia 1976:186.

Goin, C.

J.

and C. G. Jackson. 1965. Haemoglobinvalues of some amphibians and reptiles from Florida.

Herpetologica 21:1 45-146.

Haggag, G., and et al. 1966. Hibernation in reptiles II.
Changes in blood glucose, haemoglobin, red blood
cells count, protein and nonprotein nitrogen. Compar-

ative Biochemistry and Physiology. 17:335-339.

Hartman, F. A., and M. A. Lessler. 1964. Erythrocyte
measurements in fishes, amphibians and reptiles. Biological Bulletin. 126:83-88.

Heady, J. M. and T. E. Rogers. 1963. Turtle blood cellmorphology. Proceedings of the Iowa Academy ofSciences. 69:587-590.

Hutchinson,V H. and H. Szarski. 1965. Number oferythrocytes in some amphibians and reptiles. Copeia

1965:373-375.

Literature Cited
Altland, P. D. and E. C. Thompson. 1958. Some fac-Hutton, K. E. 1960. Seasonal physiological changes
in the red eared turtle Pseudemys scripta elegans.

tors affecting blood formation in turtles.

Copeia 1960:360-362.

for the Society of Experimental

Proceedings

Biology and Medi-Hutton, K. E. 1961. Blood volume, corpuscular con-cine. 99:456-459.

stants and shell weight in turtles. American Journal ofBaker, E. G. S. and L. E. Kline. 1932. Comparative
erythrocyte count of representative vertebrates. Pro-Physiology 200:1004-1006.

ceedings of the Indiana Academy of Sciences 41:417418.
Canfield, P. J. and G. M. Shea. 1988. Morphologicalobservations on the erythrocytes, leukocytes andNewlin, M. E. and R. E. Ballinger. 1976. Blood haemoglobin concentration in four species of lizards.

Copeia 1976:392-394.
Saint Girons, M.

C. 1970. Morphology of the circu-lating blood cells. Pp. 73-91. In Gans (ed.), Biology of2001

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Morphology C. Academic Press,New York.
Szarski, H. and G. Czopek. 1966. Erythrocyte diamein some amphibians and reptiles. Bulletin del'Academie Polonaise des Science. Classe 2. Serie dester

Sciences Biologiques. 14(6):437-443.
Taylor, K. and H. M. Kaplan. 1961. Light microscopyof the blood cells of Pseudemyd Turtles. Herpetologica 17:186-192.

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