ZOBODAT - www.zobodat.at
Zoologisch-Botanische Datenbank/Zoological-Botanical Database
Digitale Literatur/Digital Literature
Zeitschrift/Journal: Herpetozoa
Jahr/Year: 2017
Band/Volume: 30_1_2
Autor(en)/Author(s): Uca Onur, Arikan Hüseyin, Cicek Kerim
Artikel/Article: Blood cell morphology of Turkish gekkonid lizards (Squamata: Sauria: Gekkonidae, Phyllodactylidae) 29-37 uca_etal_BloodCellmorphology_turkish_geckos:herpetoZoA.qxd 07.08.2017 18:12 seite 1
herpetoZoA 30 (1/2): 29 - 37 29 Wien, 30. Juli 2017
Blood cell morphology of turkish gekkonid lizards (squamata: sauria: Gekkonidae, phyllodactylidae)
Blutzellmorphologie türkischer Geckos (squamata: sauria: Gekkonidae, phyllodactylidae)
onur uCA & h üseyin ArikAn & k erim ÇiÇek
kurZFAssunG Die Autoren erhoben Daten zur Größe peripherer Blutzellen (erythrozyten, Leucozyten, thrombozyten) aus mit Wright-Färbung behandelten Abstrichen für sechs Arten türkische Geckos: Asaccus barani torki , AhmADZADeh , i LGAZ , A vCi & k umLutAs , 2011 , Mediodactylus heterocercus (B LAnForD , 1874) , Mediodactylus kotschyi (s teinDAChner , 1870) , Cyrtopodion scabrum (h eyDen , 1827) , Hemidactylus turcicus (L innAeus , 1758) und Stenodactylus grandiceps hAAs , 1952 . Die längsten erythrozyten fanden sich bei S. grandiceps , die breitesten und größten bei A. barani , die kür - zesten bei C. scabrum , die schmalsten bei M. kotschyi und die kleinsten bei C. scabrum . Die längsten und größ - ten erythrozytenkerne wurden bei M. heterocercus , die kürzesten, schmalsten und kleinsten bei M. kotschyi und die größten bei A. barani gemessen . Bezüglich des verhältnisses von Länge zu Breite, kamen die länglichsten erythrozyten bei S. grandiceps , die rundlichsten bei M. kotschyi vor. Die kern-plasma-relation zeigte bei C. sca - brum und M. heterocercus den größten, bei S. grandiceps den geringsten Wert. Die kern-plasma-relation türki - scher Geckos war ähnlich der bei türkischen Lacertiden. Zwischen den untersuchten Geckoarten bestanden beträchtliche unterschiede in der Gestalt der erythrozyten und ihrer kerne, während das für die Leukozytenmorphologie nicht zutraf. Lymphozyten waren die häufigsten Leukozyten bei allen Arten. Die kerne der eosinophilen und basophilen Granulozyten waren durch die starke körnung des Zytoplasmas undeutlich. Die thrombozyten waren bei allen Arten flach ellipsoidisch. ABstrACt size parameters of peripheral blood cells (erythrocytes, leucocytes, thrombocytes) obtained from blood smears stained with Wright’s stain are given for six species of turkish gekkonid lizards: Asaccus barani torki , AhmADZADeh , i LGAZ , A vCi & k umLutAs , 2011 , Mediodactylus heterocercus (B LAnForD , 1874) , Mediodactylus kotschyi (s teinDAChner , 1870) , Cyrtopodion scabrum (h eyDen , 1827) , Hemidactylus turcicus (L innAeus , 1758) and Stenodactylus grandiceps hAAs , 1952 . the longest erythrocytes were found in S. grandiceps , the widest and largest in A. barani , the shortest in C. scabrum, the narrowest in M. kotschyi and the smallest in C. scabrum . the longest and biggest erythrocyte nuclei were measured in M. heterocercus , the shortest, narrowest and smallest in M. kotschyi and the largest in A. barani. regarding their length-to-width ratio, the most oblong erythrocytes were found in S. grandiceps , the most roundish in M. kotschyi . the value of the nucleocytoplasmic ratio was highest in C. scabrum and M. heterocercus , and low - est in S. grandiceps . the nucleocytoplasmic ratio of the gekkonid erythrocytes was similar as in lacertids. Among the species studied, erythrocyte and nucleus morphology showed considerable variation, whereas, leukocyte morphology did not. Lymphocytes outnumbered the other leucocytes in all species. the nuclei of eosinophils and basophils were not distinct because of intense granulation of the cytoplasm. the thrombocytes were the shape of a flattened ellipsoid in all species. key WorDs reptilia: squamata: sauria: phyllodactylidae, Asaccus barani ; Gekkonidae, Cyrtopodion scabrum , Hemidactylus turcicus , Mediodactylus heterocercus , Mediodactylus kotschyi , Stenodactylus grandiceps , blood smears, blood cell morphology, physiology, turkey
introDuCtion
Blood analyses are widely used tools help to indicate ecosystem health ( Deem et in the diagnosis and monitoring of animal al. 2006). the cellular metabolism, its homeo- health and disease and the differentiation of static control mechanisms and cellular con - physiological processes ( Christopher et al. figuration are optimized for specific require - 1999). these techniques are applied with ments and temperature ranges. As a result, several wildlife species, especially for the composition of blood varies consider ably threatened or endangered populations, and among reptile groups ( DessAurer 1970). uca_etal_BloodCellmorphology_turkish_geckos:herpetoZoA.qxd 07.08.2017 18:12 seite 2
30 o. u CA & h. A rikAn & k. Ç iÇek
the studies on the comparative mor - (sevinÇ et al. 2000; sevinÇ & u ğurtAŞ 2001; phology of peripheral blood cells (erythro - AtAtür et al. 2001; ArikAn et al. 2009b; Ari- cytes, leucocytes, thrombocytes) in reptiles kAn & Ç iÇek 2010), however, knowledge mainly concentrated on the seasonal and on gekkonids is still limited. in turkey, these sexual variation of counts (e.g., sZArski saurians are, according to current knowledge 1968; D essAurer 1970; D uGuy 1970) and (BArAn & A tAtür 1998; u etZ & h ošek size ( WintroBe 1933; h ArtmAn & L essLer 2016), represented by two families (phyl- 1964; s Aint Girons & s Aint Girons 1969; lodactylidae and Gekkoni dae), five genera sAint Girons 1970; s evinÇ et al. 2000; and six species [ Asaccus barani torki , A h- sevinÇ & u ğurtAŞ 2001; A tAtür et al. mADZADeh , i LGAZ , A vCi & k um LutAŞ , 2011, 2001; ArikAn et al. 2009; ArikAn & Ç iÇek Mediodactylus heterocercus (BLAnForD , 2010; A rikAn & Ç iÇek 2014 ), and the diag - 1874), Mediodactylus kotschyi (steinDACh- nosis of blood parasites ( roCA & G ALD όn ner , 1870), Cyrtopodion scabrum (heyDen , 2010). nevertheless, only a few studies 1827), Hemidactylus turcicus (LinnAeus , described the morphology and size of circu - 1758), Stenodactylus grandiceps hAAs , lating blood cells of one or more species 1952]. the objective of the pre sent study (e.g., hArtmAn & L essLer 1964; s ZArski was to obtain and discuss comparative in- & C Zopek 1966; k notková et al. 2002). formation on morphology and size of peri - there are various studies on the com - pheral blood cell types in these six gekkonid parative morphology of the peripheral blood species from turkey. cells in turkish lacertids and scincids
mAteriALs AnD methoDs
the study sample comprises a total of lengths (eL), widths (eW) nuclear lengths sixteen specimens collected in April and (nL) and nuclear widths (nW) measured. may 2000 to 2010 at various localities in the volumes of erythrocytes (ev) and their turkey (table 1). sexes were not distin - nuclei (nv) were calculated according to guished. the blood samples were obtained the formulae ev = (eL*eW 2)*( π/ 6) [µm 3] according to mAC LeAn et al. (1973), from and nv = (nL*nW 2)*( π/ 6) [µm 3]. the cel - postorbital sinuses using heparinized glass lular and nuclear shapes were compared capillaries. three smears per sample were using the eL/eW and nL/nW ratios while measured and evaluated individually. the comparison of the nucleocytoplasmic the blood smears prepared were ratios (nr) were calculated according to the stained with Wright’s stain. the cells were formula nr = nv/(ev-nv). Besides, mea - measured under a light microscope, using a surements of the largely spherical leuko - moB-1-15x Lomo eyepiece ocular micro- cytes (lymphocytes, monocytes, hetero - meter. From each blood smear, 40 erythro - phils, eosinophils, ba sophils) and thrombo - cytes were randomly chosen and their cytes (length and width; tL, tW) were made.
table 1: Collecting localities of the turkish gekkonid specimens used in the present comparative hemato - logical study. n – number of individuals. tab. 1: Fundorte der türkischen Geckos, die zur vorliegenden hämatologischen untersuchung verwendet wurden. n – Anzahl der individuen.
species / Art n Locality / Fundort Latitude / geogr. Breite Longitude / geogr. Länge Asaccus barani 2 Birecik-urfa 37.025002 37.976955 Mediodactylus heterocercus 2 mardin 37.301906 40.730414 Mediodactylus kotschyi 4 halfeti-Şanlıurfa 37.247000 37.866667 Cyrtopodion scabrum 2 Şanlıurfa 37.120305 38.784801 Hemidactylus turcicus 4 mut-mersin 36.644337 33.435555 Stenodactylus grandiceps 2 kilis 36.718399 37.121220 uca_etal_BloodCellmorphology_turkish_geckos:herpetoZoA.qxd 07.08.2017 18:12 seite 3
Blood cell morphology of turkish gekkonid lizards 31
the photographs of the blood cells were al blood cell morphology among species. taken with an olympus CX21-Altra 20 soft the alpha level was set at 0.05, and the imaging so lutions system. mean values were provided with their stan - since the measured data was normally dard deviations. All statistical analyses distributed (kolmogorov-smirnov D test, P were performed using pAst statistical ≥ 0.05), parametric one-way AnovA was package ( hAmmer et al. 2001). used to determine the variation of peripher -
resuLts
the typical shape of erythrocytes in the ratio nL/nW was considered, the most gekkonid lizards is oval, as is the case with elongate nuclei were found in S. grandiceps fish, amphibians and other reptile spe cies. and the most rounded ones in A. barani their nuclei are also oval and almost locat - (FnL/nW = 57.48, p < 0.01). the nucleocy - ed at the centre of the cell. in Wright- toplasmic ratio was highest in C. scabrum stained preparations, cytoplasm looks light and M. heterocercus , and lowest in S. yellowish pink and chromophilic nuclei are grandiceps (Fnr = 99.75, p < 0.01; table 2) . dark purplish blue (Figs. 1A, 1B, 1C). regarding leucocytes, both small and in the blood preparations considerable large lymphocytes were observed as the variation regarding length, width and volume dominant cells in the blood smears of all of the erythrocytes was detected among spe- gekkonid species. Agranulocytes (lympho - cies and even conspecifics . potential differ - cytes and monocytes) constituted 80 % of ences in the physiological status, age or sex the leucocytes of the examined species. the may have caused the observed high degree average diameter of small lymphocytes was of variation among conspecifics. For the ery - biggest (10.34 μm) in S. grandiceps and throcyte measurements, see table 2. smallest (7.11 μm) in C. scabrum (table 3). the averages of the individual species’ in small lymphocytes, the spherical nuclei erythrocyte length, width, volume, and eL/ were more chromophilic and localized in a eW ratio were ranged from 14.83 - 18.62 certain cell zone than in large lymphocytes μm, 8.34 - 9.94 μm, 541.83 - 924.37 μm3, in which the cytoplasm covered a larger and 1.72 - 2.00, respectively. the longest area and was stained a pale blue, whereas, erythrocytes were found in S. grandiceps nuclei were stained a purplish blue with (Fig. 1C, one-way AnovA, FeL = 72.20, p < Wright’s stain (Figs. 1D, 1e). the average 0.01); the widest and largest in A. barani diameter of large lymphocytes was big gest (FeW = 28.59, p < 0.01; Fes = 58.54, p < (15.18 μm) in S. grandiceps , and small est 0.01), the shortest in C. scabrum , the narrow - (10.71 μm) in C. scabrum (table 3). est in M. kotschyi and the smallest in C. monocytes resembled large lympho - scabrum. in terms of the eL/eW ratio, the cytes from which they were easily differen - most elongate erythrocytes were found in S. tiated by their kidney-shaped nuclei. Cyto- grandiceps , the most rounded ones in M. plasm was stained a light gray, and the kotschyi (table 2, FeL/eW = 16.35, p < 0.01). nuclei a dark purplish blue with Wright’s the averages of the individual species’ stain (Fig. 1F). the average monocyte dia- nuclear length, width, and volume were meter was biggest in M. heterocercus (14.58 ranged from 6.66 - 7.57 μm, 3.69 - 4.59 μm µm), and smallest (10.50 µm) in S. grandi - and 52.78 - 83.48 μm3, respectively, the mean cep s and M. kotschyi (table 3). length-to-width ratios (nL/nW) from 1.52 - Among the granulocytes, heterophils 2.05. the species’ mean nucleocytoplasmic had light blue cytoplasm and numerous gran- ratios rang ed between 0.07 - 0.15. the lon- ules of ellipsoidal shape . their bilobate gest and biggest nuclei were measured in M. nuclei were stained a dark purplish blue heterocercus (FnL = 17.93, p < 0.01; Fnv = color with Wright’s stain (Fig. 1G, 1h). the 41.33, p < 0.01 ); the shortest, narrowest mean dia meter of heterophil granulocytes (FnW = 58.81, p < 0.01) and smallest in M. was biggest (12.44 µm) in S. grandiceps and kotschyi and the largest in A. barani. When smallest (11.00 µm) in M. kotschyi (table 3). uca_etal_BloodCellmorphology_turkish_geckos:herpetoZoA.qxd 07.08.2017 18:12 seite 4
32 o. u CA & h. A rikAn & k. Ç iÇek
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Blood cell morphology of turkish gekkonid lizards 33
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34 o. u CA & h. A rikAn & k. Ç iÇek
Fig. 1: For legend see opposite page. / Abb. 1: Legende siehe gegenüberliegende seite. uca_etal_BloodCellmorphology_turkish_geckos:herpetoZoA.qxd 07.08.2017 18:12 seite 7
Blood cell morphology of turkish gekkonid lizards 35
the cytoplasm of eosinophil granulo - which masked the nucleus just like in the cytes was stained a light yellowish color eosinophils (Fig. 1J). the mean diameter of with Wright’s stain. since the nucleus was basophils, which was smaller than of other masked by large and bright red granules in granulocytes, was biggest (10.50 µm) in H. the cytoplasm, its shape couldn’t be distin - turcicus and smallest (10.00 µm) in C. guished clearly (Fig. 1i). the mean diame - scabrum (table 4). ter of eosinophils was biggest (10.75 µm) in thrombocytes were of flattened ellipsoi- H. turcicus and smallest (8.96 µm) in M. dal shape, their nuclei filling nearly the whole kotschyi (table 3). cell (Figs. 1k, 1L). the longest and largest the cytoplasm of the basophil granu - thrombocytes were observed in A. barani , locytes was filled with black granules, the shortest and narrowest in H. turcicus .
DisCussion
reptilian erythrocytes are similar in elegans ménétries , 1832 . in terms of the function and appearance to the birds’ red eL/eW ratio, the most elongate cells were blood cells but differ in size and number. determined in Lacerta pamphylica sChmiDt - Among reptiles, the largest erythrocytes are Ler , 1975, and the most spherical in Anato- observed in Sphenodon punctatus GrAy , lolacerta danfordi (G ünther , 1876) . 1831, turtles and crocodiles, the smallest in As regards reptiles from turkey, the lacertid lizards ( hArtmAn & L essLer 1964; erythrocyte and nucleus volumes observed sAint Girons & s Aint Girons 1969; s Aint by ArikAn & Ç iÇek (2010, 2014) were Girons 1970; s evinÇ et al. 2000). this het - 366.88 - 675.01 µm 3 and 46.15 - 68.50 µm 3 erogeneity in blood cell morphology is not for lacertids, and 421.52 - 497.98 µm 3 and only considerable between orders but even 16.15 - 26.72 µm 3 for scincids; the nucleo - within families (e.g., hArtmAn & L essLer cytoplasmic ratios ranged between 0.04-0.05 1964; s ZArski & C Zopek 1966; s Aint Gi- in scincids and 0.10-0.13 in lacertids. rons & s Aint Girons 1969; s Aint Girons in the present study, the largest ery - 1970; A rikAn et al. 2009a, 2009b, 2010). throcytes were seen in Asaccus barani , the ArikAn & Ç iÇek (2010) studied the smallest in Cyrtopodium scabrum , the most morphology of peripheral blood cells of var - elongate in Stenodactylus grandiceps , the ious turkish reptilian species, noted that most spherical in Mediodactylus kotschyi . there is considerable variation in the size of the mean values of the nucleo cyto - erythrocytes among or within lizard families plasmic ratios were between 0.07 and 0.15 and hypothesized that erythrocyte size might for gekkonids. Concerning the nucleocyto - be correlated with differences in the species- plasmic ratio, the gekkonids studied resem - specific activity levels. these au thors found bled the lacertids. the largest erythrocytes in Varanus griseus Lymhocytes were the dominant cells (D AuDin , 1803) , the smallest in Ophisops in the blood smears of many reptile species
______Fig. 1 (opposite page) / Abb. 1 (gegenüberliegende seite): photomicrographs of erythrocytes, leukocytes and thrombocytes of some turkish geckos . erythrocytes: A – A. barani , B – C. scabrum , C – S. grandiceps . Leukocytes: D – small lymphocyte (M. heterocercus ), e – large lymphocyte ( C. scabrum ), F – monocyte ( H. turcicus ). Granulocytes: G – hetero- phil ( C. scabrum ), h – heterophil ( C. scabrum ), i – eosinophil ( C.scabrum ), J – basophil ( C.scabrum ). thrombocyte: k – ( M. kotschyi ), L – an aggregate of thrombocytes ( S. grandiceps ). horizontal bar: 20 μm. mikroskopische Aufnahmen von erythrozyten, Leukozyten und thrombozyten einiger türkischer Geckoarten. erythrozyten: A – A. barani , B – C. scabrum , C – S. grandiceps . Leukozyten: D – kleiner Lymphozyt ( M. heterocercus ), e – großer Lymphozyt ( C. scabrum ), F – monozyt ( H. turcicus ). Granulozyten: G – heterophiler ( C.scabrum ), h – heterophiler ( C. scabrum ), i – eosinophiler ( C. scabrum ), J – basophiler ( C. scabrum ). thrombozyt: k – M. kotschyi , L – ein Aggregat von thrombozyten (S. grandiceps ). Balkenlänge entspricht 20 μm. uca_etal_BloodCellmorphology_turkish_geckos:herpetoZoA.qxd 07.08.2017 18:12 seite 8
36 o. u CA & h. A rikAn & k. Ç iÇek
(e.g., ArikAn et al. 2004, 2009a, 2010), and ArikAn et al. 2004, 2009a, 2010), which is the nuclei of eosinophils and basophils were in full agreement with the observations not easily distinguished as they were mas- made here for six gecko species. the lar- ked by dense granulation in the cytoplasm. gest thrombocytes were observed in A. ba- identical results were obtained for six geck - rani , the smallest in H. turcicus . onid species in the present study. in conclusion, the present data details reptilian thrombocytes appear as flat - the considerable morphological variation of tened ellipsoidal cells with pale cytoplasm peripheral blood cells of six gekkonid spe - and centrally localized, extremely chromo - cies occurring in turkey. Considering the philic nucleus which largely fills the cell nucleocytoplasmic ratio, the gekkonids (sAint Girons 1970; C AnFieLD & s heA 1988; were more similar to lacertids than scincids.
ACknoWLeDGments
the manuscript of this paper is based partly on inhabiting turkey. ege university, Graduate school of the msc thesis of the first author, onur uCA (u CA , o. natural and Applied sciences, izmir, turkey, pp. 16 (in 2015: Blood cell morphology of Gekkonidae species turkish).
reFerenCes
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DAte oF suBmission: march 14, 2016 Corresponding editor: heinz Grillitsch
Authors: onur uCA , hüseyin ArikAn & kerim ÇiÇek (Corresponding author < kerim.cicek@hotmail. com > or < [email protected] >) ‒ ege university, Faculty of science, Biology Department, Zoology section, tr-35100, Bornova-izmir, turkey.