© Firenze University Press www.fupress.com/ah Acta Herpetologica 5(2): 179-198, 2010 Morphology of peripheral blood cells from various species of Turkish Herpetofauna Hüseyin Arıkan, Kerim Çiçek* Ege University, Faculty of Science, Biology Department, Zoology Section, 35100, Bornova, Izmir-Tur- key. *Corresponding author. E-mail: kerim.cicek@ege.edu.tr Submitted on: 2010, 15th June; revised on: 2010, 6th October; accepted on: 2010, 27th October. Abstract. In this study, measurements of morphological and size parameters of peripheral blood cells (erythrocyte, leucocyte, thrombocyte) on blood smear prepara- tion devices stained with Wright’s stain were given for 87 species from Turkish her- petofauna (19 amphibian species including 7 urodeles and 12 anurans as well as 68 reptile species including 4 turtles, 30 lizards and 34 snakes). It was determined that erythrocyte and nucleus sizes showed great variations among the species of herpetofauna and even among the preparations of the same species; the largest blood cells (erythrocyte, leucocyte, thrombocyte) were found in urodeles; aquatic and semiaquatic species had larger erythrocytes than terrestrials, and the lar- gest erythrocytes were in turtles among the reptile species examined. Lymphocytes were determined as the predominant cells among the blood leucocytes in blood smears of all the examined species. Keywords. Amphibians, reptiles, blood smears, blood cell morphology. INTRODUCTION Blood analyses are useful, widely used tools that aid in the diagnosis and monitoring of animal health and disease and in the differentiation of physiologic processes (Christopher et al., 1999). These techniques are used with several wildlife species, especially for threat- ened or endangered populations, and help to indicate ecosystem health (Deem et al., 2006). However, much of our knowledge regarding vertebrate blood and blood cells is based on mammalian references (Claver and Quaglia, 2009). The studies of nonmammalian verte- brate blood is relatively new (e.g., Canfield, 1998; Mader, 2000; Campbell, 2004; Allander and Fry, 2008), and blood cell morphology of reptiles still completely unknown (e.g., Frye, 1991; Mader, 2000; Campbell, 2004; Strik et al., 2007, Sykes and Klaphake, 2008). The studies on the comparative morphologies of the peripheral blood cells in different amphibians and reptiles mainly concentrate on seasonal and sexual variations of counts (Vernberg, 1955; Altman and Dittmer, 1961; Foxon, 1964; Hutchison and Szaski, 1965; 180 H. Arıkan and K. Çiçek Dessaurer, 1970; Duguy, 1970; Jerrett and Mays, 1973; Alleman et al., 1999; Wojtaszek and Adamowicz, 2003; Solís et al., 2007) and sizes of blood cells (erythrocyte, leucocyte, thrombocyte) (Gulliver, 1875; Wintrobe, 1933; Hartman and Lessler, 1964; Szarski, 1968; Saint Girons and Saint Girons, 1969; Saint Girons, 1970; Wojtaszek et al., 1997; Harr et al., 2001; Knotková et al., 2002; Salakij et al., 2002), and blood parasites (Espinosa-Avilés et al., 2008; Roca and Galdón, 2010). There are several review on morphology of blood cells in amphibians and reptiles (e.g., Hutchison and Szaski, 1965; Hartman and Lessler, 1964; Szarski and Czopek, 1966; Canfield, 1998; Mader, 2000; Campbell, 2004; Allander and Fry, 2008, Sykes and Klaphake, 2008). Since 1989, there have been several studies on morphology of peripheral blood cells in Turkish amphibians (e.g., Arıkan, 1989; Atatür et al., 1998, 1999; Arıkan et al., 2001, 2003a, 2003b, 2010; Gül and Tok, 2009) and reptiles (e.g., Arıkan et al., 2004, 2009a, 2009b; Atatür et al., 2001; Sevinç et al., 2000; Uğurtaş et al., 2003). The objective of the present study is to obtain detailed information on morphology and size of peripheral blood cell in 87 amphibian and reptile species in Turkey comparatively, and the results were discussed with literature. MATERIAL AND METHODS Individuals of 87 species belonging to sexually-mature amphibians and reptiles were collected from Anatolian and Thracian parts of Turkey (Table 1). The field studies were carried out in April- May for amphibians and in April-June for reptiles. The individuals were primarily collected on sev- eral herpetofaunal trips of previous studies or projects performed between 1989 and 2009. Blood samples were obtained from heart ventriculus of amphibians via heparinized glass capillaries, from caudal vein of turtles via heparinized injector (Hutchison and Szarski, 1965; Szarski and Czopek, 1966), and from postorbital sinuses of lizard and snake individual via heparinized glass capillaries according to MacLean et al. (1973). After obtaining blood samples in reptiles, they were released to their natural environments. For each individual, approximately 4-5 blood smears were prepared and stained with Wright’s stain for the measurements of morphology and size parameters of blood cells. Blood cells were Table 1. Collecting localities of 87 species from Turkish Herpetofauna [n: number of individuals]. Species n Locality Latitude Longitude Urodela Lissotriton vulgaris 5 Bornova – Izmir 38.472031 27.262555 Lyciasalamandra atifi 4 Alanya – Antalya 36.594658 32.123258 Mertensiella caucasica 3 Akçaabat – Trabzon 41.000657 39.569261 Neurergus strauchii 10 Yam village – Bitlis 38.375950 42.091056 Ommatotriton vittatus 5 Mezitli – Mersin 36.857369 34.397636 Salamandra infraimmaculata 4 Harbiye – Antakya (Hatay) 36.138361 36.143428 Triturus karelinii 4 Kalecik - Ankara 40.097222 33.408333 (continued). 181Blood cell morphology of Turkish herpetofauna Species n Locality Latitude Longitude Anura Pelophylax bedriagae 5 Bornova-Izmir 38.472031 27.262555 Pelophylax caralitanus 10 Beyşehir – Konya 37.676389 31.726111 Rana dalmatina 4 Belgrad forest - Istanbul 41.194309 28.951383 Rana holtzi 5 Mountain Bolkar – Niğde 37.438813 34.604279 Rana macrocnemis 6 Mountain Uludağ – Bursa 40.072066 29.216721 Bufo bufo 7 Marmaris – Muğla 36.863411 28.275040 Pseudepidalea variabilis 10 Sülüklüpınar – Adana 37.040136 37.040136 Pelobates syriacus 8 Seydişehir – Konya 37.423871 31.850475 Pelodytes caucasicus 10 Uzungöl – Trabzon 40.622109 40.285267 Bombina bombina 5 Büyükdöllük – Edirne 41.760133 26.603753 Hyla arborea 5 Fethiye – Muğla 36.651389 29.123056 Hyla savignyi 6 Alanyalı village – Mersin 37.094734 34.501284 Testudines Emys orbicularis 10 Lake Yayla – Denizli 38.059675 28.778826 Mauremys caspica 8 Nusaybin – Mardin 37.066667 41.216667 Mauremys rivulata 3 Northern Cyprus 35.237616 33.471477 Testudo graeca 8 Izmir 38.418850 27.128720 Lacertilia Ablepharus chernovi 4 Çamardı – Niğde 37.832029 34.986486 Chalcides ocellatus 6 Finike – Antalya 36.300827 30.144497 Eumeces schneideri 5 Meke saltern – Konya 37.682887 33.636460 Ophiomorus punctatissimus 4 Kaş – Antalya 36.204441 29.638982 Trachylepis aurata 4 Karapınar – Konya 37.714821 33.552237 Trachylepis vittata 4 Kırobası – Mersin 36.722014 33.909358 Acanthodactylus boskianus 4 Adana 36.999996 35.321314 Acanthodactylus harranensis 7 Şanlıurfa 37.149994 38.799857 Anatololacerta danfordi 4 Çamlıyayla - Mersin 37.170139 34.608260 Apathya cappadocica 8 Ulukışla – Niğde 38.055150 34.310216 Darevskia praticola 4 Kırklareli 41.733333 27.216667 Darevskia uzzelli 5 Kars 40.592680 43.077692 Darevskia valentini 12 Ardahan 41.110477 42.702174 Lacerta pamphylica 3 Mut – Mersin 36.644337 33.435555 Lacerta trilineata 7 Çamlıyayla – Mersin 37.170139 34.608260 Lacerta viridis 4 Hendek – Adapazarı (Sakarya) 40.805100 30.749291 Mesalina brevirostris 2 Akçakale – Şanlıurfa 36.711006 38.947988 Ophisops elegans 6 Mut – Mersin 36.644337 33.435555 Parvilacerta parva 2 Çamardı – Niğde 37.832029 34.986486 Podarcis muralis 2 Kırklareli 41.733333 27.216667 Podarcis siculus 2 Istanbul 41.005270 28.976960 Timon princes 2 Eruh – Siirt 37.750000 42.183333 Eublepharis angramainyu 3 Birecik – Şanlıurfa 37.025002 37.976955 Table 1. (continued). (continued). 182 H. Arıkan and K. Çiçek Species n Locality Latitude Longitude Cyrtopodion heterocercum 2 Mardin 37.301906 40.730414 Cyrtopodion scabrum 2 Şanlıurfa 37.120305 38.784801 Hemidactylus turcicus 3 Northern Cyprus 35.237616 33.471477 Laudakia stellio 2 Mut – Mersin 36.644337 33.435555 Trapelus lessonae 2 Birecik – Şanlıurfa 37.025002 37.976955 Chamaeleo chamaeleon 2 Northern Cyprus 35.237616 33.471477 Varanus griseus 1 Viranşehir – Şanlıurfa 37.178374 39.761510 Serpentes Leptotyphlops macrorhynchus 2 Birecik – Şanlıurfa 37.025002 37.976955 Typhlops vermicularis 5 Mut – Mersin 36.644337 33.435555 Eryx jaculus 2 Mut – Mersin 36.644337 33.435555 Dolichophis caspius 2 Ulukışla – Niğde 38.055150 34.310216 Dolichophis jugularis 2 Mut – Mersin 36.644337 33.435555 Dolichophis schmidti 2 Suruç – Şanlıurfa 36.974652 38.424516 Eirenis barani 1 Kahramanmaraş 37.583309 36.933403 Eirenis coronella 2 Birecik – Şanlıurfa 37.025002 37.976955 Eirenis decemlineatus 1 Diyarbakır 37.914409 40.230624 Eirenis eiselti 2 Diyarbakır 37.914409 40.230624 Eirenis levantinus 1 Samandağ – Antakya (Hatay) 36.082392 35.999324 Eirenis modestus 2 Çamlıyayla - Mersin 37.170139 34.608260 Eirenis punctatolineatus 2 Eruh – Siirt 37.750000 42.183333 Eirenis rothii 2 Küplüce – Kilis 36.757230 37.237016 Hemorrhois nummifer 2 Mut – Mersin 36.644337 33.435555 Hemorrhois ravergieri 1 Sakçagözü - Gaziantep 36.715370 37.117360 Malpolon monspessulanus 2 Çiğli – İzmir 38.499432 27.038216 Natrix natrix 4 Mut – Mersin 36.644337 33.435555 Natrix tessellata 4 Beyşehir – Konya 37.676389 31.726111 Platyceps collaris 1 Midyat – Mardin 37.416667 41.369719 Platyceps najadum 1 Mut – Mersin 36.644337 33.435555 Platyceps ventromaculatus 1 Harran – Şanlıurfa 36.866667 39.033331 Rhynchocalamus melanocephalus 1 Antakya (Hatay) 36.401829 36.349788 Spalerosophis diadema 2 Birecik – Şanlıurfa 37.025002 37.976955 Telescopus fallax 2 Northern Cyprus 35.237616 33.471477 Telescopus nigriceps 2 Kilis 36.718399 37.121220 Zamenis hohenackeri 2 Antakya (Hatay) 36.401829 36.349788 Zamenis longissimus 1 Zonguldak 41.456406 31.798752 Macrovipera lebetina 2 Dikmen – Northern Cyprus 35.268159 33.324760 Montivipera albizona 1 Mountain Balık – Kahramanmaraş 37.516405 36.449976 Montivipera wagneri 2 Karakurt – Kars 40.169027 42.605943 Montivipera xanthina 2 Gümüldür – Izmir 38.076415 27.022031 Vipera eriwanensis 2 Ardahan 41.110477 42.702174 Walterinnesia morgani 1 Tek Tek Mountains – Şanlıurfa 36.812953 39.252838 Table 1. (continued). 183Blood cell morphology of Turkish herpetofauna measured using a MOB-1-15× micrometrical ocular. Lengths (L) and widths (W) of 40 random- ly chosen erythrocytes as well as nuclear lengths (NL) and nuclear widths (NW) were measured for each blood smear. Erythrocyte sizes (ES) and their nuclei sizes (NS) were computed from ES= LWπ/4 and NS= NLNWπ/4. Cells and nuclear shapes were compared with L/W and NL/NW ratios, and nucleus/cytoplasm with NS/ES ratio. In addition, from the blood smears of each species, meas- urements of leucocytes (lymphocytes, monocytes, heterophils, eosinophils, basophils) and thrombo- cytes (TL, TW) were also taken to determine their sizes. The photomicrographs of the blood cells were taken with Olympus BX51-Altra 20 Soft Imaging System. Correlation between body size and erythrocyte size were analyzed by non-parametric kendall τ test. RESULTS Characteristic erythrocyte shape of amphibians and reptiles we analyses is oval, simi- lar to that of vertebrate fish and birds. Except for Montivipera xanthina, the erythrocytes of the examined species have a somewhat ellipsoidal nucleus, uniformly located in the centre of the cell (Fig. 1A). However, in M. xanthina, the irregularly shaped nuclei were determined in erythrocytes (Fig. 1L). On smears stained with Wright’s stain, the cyto- plasms were light yellowish pink and the chromophilic nuclei were dark purplish blue. The blood smears of the examined species demonstrated interspecific and even intraspecific variations in terms of the lengths, widths and sizes of the erythrocytes and nuclei. The erythrocyte measurements (lengths and widths), sizes, L/W ratios, nuclear measurements and nucleocytoplasmic ratios are given in Table 2. Among the amphibian and reptile species of Turkish herpetofauna, the largest eryth- rocyte was observed in urodele species (Fig. 1A). Mean length, width and size of erythro- cytes in urodeles ranged respectively between 28.06 μm-33.28 μm, 16.63 μm-20.13 μm and 367.05 μm2-523.44 μm2. In addition, L/W ratio, mean lucleus length, mean nucleus width, mean nucleus size, NL/NW ratio and nucleocytopasmic ratio (NS/ES) were found to change between 1.63-1.80, 13.86 μm-16.86 μm, 8.53 μm-10.46 μm, 92.85 μm2-138.51 μm2,1.56-1.69 and 0.22-0.34, respectively. The biggest erythrocytes and nuclei were observed in Salaman- dra infraimmaculata and the smallest in Ommatotriton vittatus. Similarly; the most strong- ly ellipsoidal erythrocytes and nuclei were observed in Mertensiella caucasica and the least ellipsoidal in Triturus karelinii. And, the shortest nucleus was observed in Lissotriton vul- garis, and the least ellipsoidal nuclei in Neurergus strauchii (Table 2). Anurans were determined to have smaller erythrocytes and nuclei than urodeles (Fig. 1B, C). Mean erythrocyte length, ranged between 15.29 μm-24.36 μm, erythrocyte width 9.68 μm-15.05 μm, erythrocyte size 116.42 μm2-276.62 μm2, L/W ratio 1.63-2.35, mean nucleus length 6.21 μm-9.59 μm, nucleus width 3.47 μm-5.03 μm, nucleus size 18.13 μm2- 36.66 μm2, NL/NW ratio 1.61-2.35 and nucleocytoplasmic ratio 0.10-0.14. Among the anuran species examined, the largest and the most strongly ellipsoidal erythrocytes were observed in aquatic Pelophylax caralitanus (Fig. 1C) and the smallest erythrocytes in ter- restrial Pelodytes caucasicus (Fig. 1C). The least ellipsoidal cells were found in Pseudep- idalea varibilis and the largest nuclei in Bombina bombina; in addition, the shortest nuclei in Pelobates syriacus, the most strongly ellipsoidal nuclei in Hyla arborea, and the least ellipsoidal nuclei in Rana dalmatina (Table 2). 184 H. Arıkan and K. Çiçek Of the reptiles, the largest erythrocytes were observed in turtles. And among the tur- tles, the largest erythrocytes were observed in aquatic species (e.g. 200.67 μm2 in Emys orbicularis), and the smallest erythrocytes in a terrestrial species Testudo greaca as 163.81 μm2 (Fig. 1D, E). In turtles; mean erythrocyte length ranged between 17.35 μm-19.99 μm, erythrocyte width 11.90 μm-12.76 μm, erythrocyte size 163.81 μm2-200.67 μm2, L/W ratio 1.47-1.61, mean nucleus length 6.09 μm-7.15 μm, nucleus width 4.91 μm-6.31 μm, nucleus size 23.60 μm2-35.64 μm2, NL/NW ratio 1.14-1.25 and nucleocytoplasmic ratio 0.15-0.20 (Table 3). In this regard; the longest, widest and largest erythrocytes were observed in E. orbicu- laris; the most strongly ellipsoidal erythrocytes and the least ellipsoidal nuclei in Maure- mys caspica. In addition, the smallest and the least ellipsoidal cells were found in T. graeca and the longest, widest and largest nuclei in M. caspica. However, the shortest, narrowest and the most strongly ellipsoidal nuclei were determined in T. graeca; the biggest nucle- Fig. 1. Photomicrographs of erythrocytes of some species belong to Turkish Herpetofauna. A: O. vittatus, B: P. caralitanus, C: P. caucasicus, D: E. orbicularis, E: T. graeca, F: O. elegans, G: M. brevirostris, H: A. danfordi, I: L. trilineata, J: L. macrorhynchus, K: H. ravergieri, L: M. xanthina. Horizontal bar: 20 μm. 185Blood cell morphology of Turkish herpetofauna Ta bl e 2. Th e er yt hr oc yt e an d th ei r nu cl ei m ea su re m en ts ( ± w ith t he ir s ta nd ar d er ro rs ) es ta bl is he d in t he p er ip he ra l b lo od s of 1 9 am ph ib ia n sp ec ie s be lo ng t o 7 fa m ili es f ro m T ur ke y [L : E ry th ro cy te le ng th , W : E ry th ro cy te w id th , E S: E ry th ro cy te s iz e, N L: N uc le us le ng th , N W : N uc le us w id th , N S: N uc le us s iz e; N S/ ES : N uc le oc yt op la sm ic r at io ]. Sp ec ie s Er yt hr oc yt es N uc le i L (μ m ) W ( μm ) L/ W ES ( μm 2 ) N L (μ m ) N W ( μm ) N L/ N W N S (μ m 2 ) N S/ ES U ro de la Sa la m an dr id ae Li ss ot ri to n vu lg ar is 30 .0 2± 0. 16 17 .8 1± 0. 08 1. 69 ±0 .0 1 41 9. 44 ±3 .1 1 13 .8 6± 0. 13 8. 53 ±0 .0 7 1. 62 ±0 .0 2 92 .8 5± 0. 68 0. 22 ±0 .0 02 Ly ci as al am an dr a at ifi 33 .2 8± 0. 17 19 .4 4± 0. 09 1. 73 ±0 .0 2 50 7. 54 ±3 .3 5 14 .9 9± 0. 11 9. 44 ±0 .0 9 1. 59 ±0 .0 2 11 1. 14 ±0 .6 4 0. 22 ±0 .0 02 M er te si el la c au ca si ca 31 .6 9± 0. 29 17 .6 9± 0. 29 1. 80 ±0 .0 2 44 0. 44 ±5 .7 9 16 .6 4± 0. 16 9. 84 ±0 .0 9 1. 69 ±0 .0 2 12 8. 60 ±0 .9 2 0. 29 ±0 .0 02 N eu re rg us s tr au ch ii 31 .2 0± 0. 21 18 .9 3± 0. 08 1. 65 ±0 .0 1 46 3. 82 ±4 .3 5 15 .4 5± 0. 07 9. 88 ±0 .0 3 1. 56 ±0 .0 3 12 0. 10 ±0 .7 7 0. 26 ±0 .0 02 O m m at ot ri to n vi tt at us 28 .0 6± 0. 16 16 .6 3± 0. 12 1. 70 ±0 .0 1 36 7. 05 ±3 .8 2 16 .0 3± 0. 14 9. 86 ±0 .0 9 1. 63 ±0 .0 2 12 4. 14 ±0 .7 0 0. 34 ±0 .0 02 Sa la m an dr a in fr ai m m ac ul at a 33 .1 0± 0. 20 20 .1 3± 0. 13 1. 65 ±0 .0 2 52 3. 44 ±4 .7 9 16 .8 6± 0. 19 10 .4 6± 0. 11 1. 61 ±0 .0 2 13 8. 51 ±0 .8 4 0. 27 ±0 .0 02 Tr itu ru s ka re lin ii 29 .5 0± 0. 16 18 .1 4± 0. 09 1. 63 ±0 .0 1 42 0. 37 ±2 .9 6 14 .9 8± 0. 11 9. 44 ±0 .0 7 1. 59 ±0 .0 3 11 1. 06 ±0 .7 4 0. 26 ±0 .0 02 A nu ra R an id ae Pe lo ph yl ax b ed ri ag ae 23 .1 4± 0. 22 13 .1 0± 0. 09 1. 61 ±0 .0 3 26 5. 63 ±2 .5 3 8. 51 ±0 .1 1 5. 01 ±0 .0 1 1. 58 ±0 .0 3 38 .3 4± 0. 74 0. 14 ±0 .0 03 Pe lo ph yl ax c ar al ita nu s 24 .3 6± 0. 23 14 .4 6± 0. 11 1. 69 ±0 .0 1 27 6. 62 ±3 .8 6 8. 13 ±0 .1 3 5. 03 ±0 .0 3 1. 61 ±0 .0 2 32 .1 5± 0. 62 0. 12 ±0 .0 01 R an a da lm at in a 19 .9 9± 0. 24 12 .1 1± 0. 11 1. 65 ±0 .0 2 19 0. 47 ±3 .5 4 8. 78 ±0 .0 9 5. 59 ±0 .0 2 1. 57 ±0 .0 2 38 .4 8± 0. 42 0. 20 ±0 .0 01 R an a ho ltz i 19 .1 0± 0. 12 12 .8 0± 0. 06 1. 64 ±0 .1 0 19 2. 81 ±1 .8 3 7. 84 ±0 .0 4 4. 13 ±0 .0 4 1. 93 ±0 .0 2 25 .4 6± 0. 27 0. 13 ±0 .0 02 R an a m ac ro cn em is 20 .5 5± 0. 12 13 .4 6± 0. 05 1. 54 ±0 .0 1 21 7. 68 ±1 .7 6 8. 66 ±0 .0 6 4. 14 ±0 .0 4 2. 14 ±0 .0 3 28 .0 3± 0. 28 0. 13 ±0 .0 02 Bu fo ni da e Bu fo b uf o 20 .8 5± 0. 10 13 .4 5± 0. 07 1. 55 ±0 .0 1 22 1. 22 ±1 .9 0 7. 81 ±0 .1 0 4. 34 ±0 .1 0 1. 86 ±0 .0 5 26 .6 0± 0. 63 0. 12 ±0 .0 02 Ps eu de pi da le a vi ri di s 17 .8 6± 0. 07 12 .7 1± 0. 04 1. 38 ±0 .0 1 17 9. 18 ±0 .9 6 6. 25 ±0 .1 3 3. 72 ±0 .0 4 1. 96 ±0 .0 5 18 .1 3± 0. 56 0. 11 ±0 .0 02 Pe lo ba tid ae Pe lo ba te s sy ri ac us 17 .5 6± 0. 08 11 .7 0± 0. 07 1. 50 ±0 .0 1 16 1. 85 ±1 .3 1 6. 63 ±0 .0 9 3. 47 ±0 .0 4 1. 96 ±0 .0 5 18 .1 3± 0. 56 0. 11 ±0 .0 02 Pe lo di tid ae Pe lo dy te s ca uc as ic us 17 .5 6± 0. 08 9. 68 ±0 .0 9 1. 58 ±0 .0 1 11 6. 42 ±2 .0 7 6. 21 ±0 .0 7 3. 81 ±0 .0 5 1. 63 ±0 .0 2 18 .7 1± 0. 31 0. 16 ±0 .0 02 B om bi na to ri da e Bo m bi na b om bi na 21 .8 0± 0. 12 15 .0 5± 0. 08 1. 45 ±0 .0 2 25 8. 14 ±2 .3 6 9. 59 ±0 .1 6 4. 88 ±0 .0 6 1. 98 ±0 .0 4 36 .6 6± 0. 82 0. 14 ±0 .0 02 H yl id ae H yl a ar bo re a 19 .8 0± 0. 10 12 .8 9± 0. 06 1. 54 ±0 .0 1 20 0. 33 ±1 .6 6 7. 94 ±0 .1 0 3. 50 ±0 .0 8 2. 35 ±0 .0 7 21 .8 8± 0. 61 0. 11 ±0 .0 01 H yl a sa vi gn yi 18 .6 3± 0. 18 12 .4 1± 0. 08 1. 50 ±0 .0 2 18 1. 44 ±1 .9 8 7. 09 ±0 .0 9 3. 97 ±0 .0 8 1. 82 ±0 .0 4 22 .3 0± 0. 55 0. 12 ±0 .0 02 186 H. Arıkan and K. Çiçek Ta bl e 3. Th e er yt hr oc yt e an d th ei r nu cl ei m ea su re m en ts ( ± w ith t he ir s ta nd ar d er ro rs ) es ta bl is he d in t he p er ip he ra l bl oo ds o f 4 tu rt le s sp ec ie s be lo ng t o 3 fa m ili es , 3 0 liz ar d sp ec ie s be lo ng to 7 fa m ili es , a nd 3 4 sn ak e sp ec ie s be lo ng to 6 fa m ili es fr om T ur ke y. Sp ec ie s Er yt hr oc yt es N uc le i L (μ m ) W ( μm ) L/ W ES ( μm 2 ) N L (μ m ) N W ( μm ) N L/ N W N S (μ m 2 ) N S/ ES Te st ud in es Em yd id ae Em ys o rb ic ul ar is 19 .9 9± 0. 11 12 .7 6± 0. 09 1. 58 ±0 .0 1 20 0. 67 ±1 .8 8 7. 15 ±0 .0 5 6. 26 ±0 .1 9 1. 19 ±0 .0 1 35 .3 7± 1. 10 0. 18 ±0 .0 1 G eo em yd id ae M au re m ys c as pi ca 18 .9 9± 0. 09 11 .9 0± 0. 07 1. 61 ±0 .0 1 17 7. 64 ±1 .4 2 7. 15 ±0 .0 5 6. 31 ±0 .0 4 1. 14 ±0 .0 1 35 .6 4± 0. 41 0. 20 ±0 .0 1 M au re m ys r iv ul at a 19 .0 2± 0. 12 12 .1 9± 0. 08 1. 57 ±0 .0 1 18 2. 74 ±1 .9 7 6. 72 ±0 .0 4 5. 89 ±0 .0 4 1. 15 ±0 .0 1 31 .2 2± 0. 35 0. 18 ±0 .0 1 Te st ud in id ae Te st ud o gr ae ca 17 .3 5± 0. 14 11 .9 6± 0. 11 1. 47 ±0 .0 1 16 3. 81 ±2 .3 4 6. 09 ±0 .0 5 4. 91 ±0 .0 4 1. 25 ±0 .0 1 23 .6 0± 0. 34 0. 15 ±0 .0 1 Sq ua m at a, S au ri a Sc in ci da e A bl ep ha ru s ch er no vi 14 .1 3± 0. 07 7. 58 ±0 .0 3 1. 87 ±0 .0 1 84 .1 2± 0. 57 6. 12 ±0 .0 5 2. 50 ±0 .0 0 2. 45 ±0 .0 2 12 .0 1± 0. 10 0. 14 ±0 .0 01 C ha lc id es o ce lla tu s 14 .6 8± 0. 06 7. 92 ±0 .0 4 1. 86 ±0 .0 1 91 .3 3± 0. 61 5. 15 ±0 .0 3 2. 64 ±0 .0 3 1. 98 ±0 .0 2 10 .7 0± 0. 13 0. 12 ±0 .0 01 Eu m ec es s ch ne id er i 15 .1 7± 0. 06 7. 74 ±0 .0 4 1. 97 ±0 .0 1 92 .3 1± 0. 56 7. 11 ±0 .0 4 2. 54 ±0 .0 2 2. 81 ±0 .0 2 14 .2 0± 0. 13 0. 15 ±0 .0 01 O ph io m or us p un ct at is si m us 15 .1 4± 0. 06 7. 73 ±0 .0 4 1. 96 ±0 .0 1 92 .0 8± 0. 64 6. 05 ±0 .0 5 2. 68 ±0 .0 4 2. 30 ±0 .0 3 12 .7 0± 0. 21 0. 14 ±0 .0 02 Tr ac hy le pi s au ra ta 14 .2 7± 0. 08 7. 56 ±0 .0 2 1. 90 ±0 .0 1 84 .8 8± 0. 54 5. 06 ±0 .0 2 2. 52 ±0 .0 1 2. 01 ±0 .0 1 10 .0 2± 0. 07 0. 12 ±0 .0 01 Tr ac hy le pi s vi tt at a 14 .1 4± 0. 08 7. 55 ±0 .0 2 1. 87 ±0 .0 1 83 .7 7± 0. 53 6. 14 ±0 .0 5 2. 50 ±0 .0 0 2. 46 ±0 .0 2 12 .0 6± 0. 10 0. 14 ±0 .0 01 La ce rt id ae A ca nt ho da ct yl us b os ki an us 14 .2 2± 0. 98 7. 92 ±0 .4 1 1. 80 ±0 .1 4 88 .4 5± 8. 37 6. 24 ±0 .5 5 4. 02 ±0 .1 2 1. 56 ±0 .1 5 19 .6 9± 1. 70 0. 23 ±0 .0 2 A ca nt ho da ct yl us h ar ra ne ns is 15 .4 6± 1. 24 8. 56 ±0 .5 9 1. 81 ±0 .1 3 10 4. 22 ±1 3. 53 6. 59 ±0 .5 0 4. 07 ±0 .1 3 1. 62 ±0 .1 5 21 .0 2± 1. 49 0. 21 ±0 .0 2 A na to lo la ce rt a da nf or di 14 .1 4± 1. 17 9. 09 ±0 .5 1 1. 56 ±0 .1 1 10 1. 13 ±1 2. 10 6. 73 ±0 .6 1 4. 41 ±0 .1 3 1. 53 ±0 .1 4 23 .3 2± 2. 30 0. 23 ±0 .0 3 A pa th ya c ap pa do ci ca 13 .4 2± 0. 90 7. 94 ±0 .4 7 1. 69 ±0 .1 4 83 .7 3± 8. 13 6. 33 ±0 .4 6 4. 11 ±0 .1 3 1. 54 ±0 .1 3 20 .4 2± 1. 58 0. 25 ±0 .0 2 (c on tin ue d) . 187Blood cell morphology of Turkish herpetofauna Sp ec ie s Er yt hr oc yt es N uc le i L (μ m ) W ( μm ) L/ W ES ( μm 2 ) N L (μ m ) N W ( μm ) N L/ N W N S (μ m 2 ) N S/ ES D ar ev sk ia p ra tic ol a 13 .0 8± 0. 97 8. 01 ±0 .3 8 1. 64 ±0 .1 3 82 .3 4± 7. 78 6. 19 ±0 .4 3 4. 31 ±0 .1 2 1. 44 ±0 .0 2 20 .9 3± 1. 46 0. 26 ±0 .0 2 D ar ev sk ia u zz el li 13 .6 5± 0. 96 7. 84 ±0 .4 8 1. 74 ±0 .1 2 84 .2 2± 9. 33 5. 98 ±0 .3 6 4. 34 ±0 .1 4 1. 38 ±0 .1 1 20 .3 6± 1. 22 0. 25 ±0 .0 3 D ar ev sk ia v al en tin i 13 .3 2± 0. 93 7. 73 ±0 .5 7 1. 73 ±0 .1 4 80 .9 7± 9. 47 6. 13 ±0 .4 8 4. 28 ±0 .1 6 1. 43 ±0 .1 0 20 .6 3± 2. 03 0. 26 ±0 .0 3 La ce rt a pa m ph yl ic a 15 .6 1± 1. 00 7. 89 ±0 .5 2 1. 99 ±0 .1 6 96 .7 7± 9. 91 6. 33 ±0 .4 1 4. 23 ±0 .1 1 1. 49 ±0 .0 9 21 .0 1± 1. 61 0. 22 ±0 .0 2 La ce rt a tr ili ne at a 14 .3 9± 1. 01 7. 63 ±0 .4 9 1. 89 ±0 .1 2 86 .3 1± 10 .2 2 6. 93 ±0 .5 2 3. 93 ±0 .1 1 1. 77 ±0 .1 6 21 .3 8± 1. 56 0. 25 ±0 .0 2 La ce rt a vi ri di s 14 .9 4± 1. 04 8. 16 ±0 .5 6 1. 83 ±0 .1 0 96 .0 3± 11 .7 8 6. 64 ±0 .5 2 4. 36 ±0 .1 3 1. 53 ±0 .1 4 22 .6 8± 1. 73 0. 24 ±0 .0 2 M es al in a br ev ir os tr is 14 .0 6± 0. 91 8. 07 ±0 .4 2 1. 75 ±0 .1 4 89 .0 9± 7. 52 6. 46 ±0 .4 9 3. 84 ±0 .1 5 1. 69 ±0 .1 7 19 .4 8± 1. 34 0. 22 ±0 .0 2 O ph is op s el eg an s 12 .4 3± 0. 65 7. 51 ±0 .2 5 1. 66 ±0 .0 9 73 .2 7± 4. 88 6. 51 ±0 .3 4 3. 84 ±0 .1 5 1. 70 ±0 .1 1 19 .6 3± 1. 22 0. 27 ±0 .0 2 Pa rv ila ce rt a pa rv a 13 .6 3± 0. 86 8. 01 ±0 .4 4 1. 70 ±0 .1 2 85 .8 0± 8. 39 6. 12 ±0 .4 8 3. 98 ±0 .1 0 1. 54 ±0 .1 2 19 .1 5± 1. 67 0. 22 ±0 .0 2 Po da rc is m ur al is 13 .9 3± 0. 95 8. 43 ±0 .5 9 1. 66 ±0 .1 1 92 .4 6± 11 .3 2 6. 36 ±0 .5 4 4. 35 ±0 .1 2 1. 46 ±0 .1 2 21 .7 4± 2. 06 0. 24 ±0 .0 2 Po da rc is s ic ul us 13 .8 9± 0. 94 8. 10 ±0 .3 5 1. 74 ±0 .1 2 87 .4 1± 7. 85 6. 59 ±0 .5 1 4. 19 ±0 .1 2 1. 57 ±0 .1 3 21 .6 9± 1. 82 0. 25 ±0 .0 2 Ti m on p ri nc ep s 14 .9 8± 1. 14 8. 43 ±0 .4 7 1. 78 ±0 .1 4 99 .2 7± 10 .8 3 6. 04 ±0 .5 3 3. 99 ±0 .1 1 1. 52 ±0 .1 5 18 .8 9± 1. 67 0. 19 ±0 .0 2 Eu pl eb ha ri da e Eu bl ep ha ri s an gr am ai ny u 16 .5 7± 0. 17 8. 93 ±0 .0 8 1. 86 ±0 .0 2 11 6. 29 ±1 .9 5 7. 38 ±0 .0 7 4. 38 ±0 .0 2 1. 69 ±0 .0 2 25 .3 5± 0. 24 0. 22 ±0 .0 1 G ek ko ni da e C yr to po di on h et er oc er cu m 16 .1 7± 0. 20 8. 81 ±0 .0 6 1. 84 ±0 .0 3 11 1. 77 ±1 .5 7 7. 57 ±0 .0 8 4. 59 ±0 .0 2 1. 65 ±0 .0 2 27 .2 7± 0. 33 0. 24 ±0 .0 1 C yr to po di on s ca br um 14 .8 3± 0. 10 8. 34 ±0 .0 7 1. 78 ±0 .0 1 97 .1 3± 1. 26 7. 08 ±0 .0 6 4. 38 ±0 .0 2 1. 62 ±0 .0 1 24 .3 4± 0. 24 0. 25 ±0 .0 1 H em id ac ty lu s tu rc ic us 16 .5 6± 0. 21 8. 91 ±0 .0 6 1. 86 ±0 .0 2 11 5. 89 ±1 .9 3 7. 44 ±0 .0 9 4. 40 ±0 .0 2 1. 69 ±0 .0 2 25 .7 1± 0. 32 0. 22 ±0 .0 1 A ga m id ae La ud ak ia s te lli o 16 .8 5± 0. 18 9. 12 ±0 .0 6 1. 85 ±0 .0 2 12 0. 71 ±1 .7 1 7. 84 ±0 .0 8 4. 40 ±0 .0 2 1. 79 ±0 .0 2 27 .0 8± 0. 29 0. 23 ±0 .0 1 Tr ap el us le ss on ae 14 .7 5± 0. 16 8. 69 ±0 .0 8 1. 70 ±0 .0 2 10 0. 78 ±1 .6 7 6. 91 ±0 .0 6 4. 58 ±0 .0 2 1. 51 ±0 .0 1 24 .8 3± 0. 25 0. 25 ±0 .0 1 Ta bl e 3. ( co nt in ue d) . (c on tin ue d) . 188 H. Arıkan and K. Çiçek Sp ec ie s Er yt hr oc yt es N uc le i L (μ m ) W ( μm ) L/ W ES ( μm 2 ) N L (μ m ) N W ( μm ) N L/ N W N S (μ m 2 ) N S/ ES C ha m ae le on id ae C ha m ae le o ch am ae le on 15 .9 7± 0. 16 9. 75 ±0 .0 8 1. 64 ±0 .0 2 12 2. 34 ±1 .8 1 7. 72 ±0 .0 9 4. 85 ±0 .0 3 1. 59 ±0 .0 2 29 .3 7± 0. 35 0. 24 ±0 .0 1 V ar an id ae Va ra nu s gr is eu s 16 .2 4± 0. 13 10 .2 1± 0. 09 1. 59 ±0 .0 1 13 0. 33 ±1 .9 9 7. 09 ±0 .0 7 4. 69 ±0 .0 3 1. 51 ±0 .0 2 26 .1 2± 0. 34 0. 20 ±0 .0 1 Sq ua m at a, O ph id ia Le pt ot yp hl op id ae Le pt ot yp hl op s m ac ro rh yn ch us 15 .8 6± 0. 11 9. 29 ±0 .0 8 1. 71 ±0 .0 2 11 5. 75 ±1 .4 5 7. 33 ±0 .0 8 4. 45 ±0 .0 2 1. 65 ±0 .0 2 25 .5 8± 0. 31 0. 22 ±0 .0 1 Ty ph lo pi da e Ty ph lo ps v er m ic ul ar is 16 .5 7± 0. 17 9. 13 ±0 .0 6 1. 82 ±0 .0 2 11 8. 76 ±1 .6 0 7. 27 ±0 .0 8 4. 54 ±0 .0 2 1. 60 ±0 .0 2 25 .9 3± 0. 29 0. 22 ±0 .0 1 B oi da e Er yx ja cu lu s 16 .3 6± 0. 19 8. 77 ±0 .0 7 1. 87 ±0 .0 2 11 2. 83 ±2 .0 1 7. 16 ±0 .0 9 4. 39 ±0 .0 2 1. 63 ±0 .0 2 24 .6 7± 0. 33 0. 22 ±0 .0 2 C ol ub ri da e D ol ic ho ph is c as pi us 14 .9 1± 0. 16 7. 64 ±0 .1 2 1. 96 ±0 .0 2 89 .8 8± 2. 21 10 .0 1± 0. 08 4. 84 ±0 .0 4 2. 07 ±0 .0 2 38 .0 8± 0. 52 0. 43 ±0 .0 1 D ol ic ho ph is ju gu la ri s 16 .2 9± 0. 18 7. 48 ±0 .0 7 2. 18 ±0 .0 2 95 .8 1± 1. 69 10 .5 7± 0. 06 4. 98 ±0 .0 4 2. 13 ±0 .0 2 41 .2 7± 0. 37 0. 44 ±0 .0 1 D ol ic ho ph is s ch m id ti 16 .2 1± 0. 14 9. 88 ±0 .0 7 1. 64 ±0 .0 1 12 5. 82 ±1 .7 1 7. 67 ±0 .0 8 4. 27 ±0 .0 2 1. 80 ±0 .0 2 25 .7 0± 0. 31 0. 20 ±0 .0 1 Ei re ni s ba ra ni 16 .1 8± 0. 12 9. 68 ±0 .0 5 1. 67 ±0 .0 1 12 2. 98 ±1 .2 7 7. 94 ±0 .0 8 4. 54 ±0 .0 2 1. 75 ±0 .0 2 28 .3 2± 0. 29 0. 23 ±0 .0 1 Ei re ni s co ro ne lla 16 .5 9± 0. 23 10 .2 2± 0. 11 1. 63 ±0 .0 2 13 3. 52 ±2 .8 1 7. 43 ±0 .0 9 4. 58 ±0 .0 2 1. 62 ±0 .0 2 26 .6 6± 0. 34 0. 20 ±0 .0 1 Ei re ni s de ce m lin ea tu s 14 .7 5± 0. 14 10 .0 3± 0. 07 1. 47 ±0 .0 1 11 6. 25 ±1 .5 5 7. 68 ±0 .1 0 4. 50 ±0 .0 3 1. 71 ±0 .0 3 27 .1 2± 0. 37 0. 23 ±0 .0 1 Ei re ni s ei se lti 14 .1 3± 0. 13 9. 62 ±0 .0 7 1. 47 ±0 .0 1 10 6. 84 ±1 .6 5 7. 29 ±0 .0 5 4. 54 ±0 .0 2 1. 60 ±0 .0 1 26 .0 0± 0. 22 0. 25 ±0 .0 1 Ei re ni s le va nt in us 16 .6 0± 0. 15 10 .0 4± 0. 09 1. 66 ±0 .0 2 13 0. 84 ±1 .7 7 8. 12 ±0 .0 8 4. 47 ±0 .0 2 1. 82 ±0 .0 2 28 .4 7± 0. 29 0. 22 ±0 .0 1 Ei re ni s m od es tu s 14 .4 7± 0. 15 7. 45 ±0 .0 8 1. 95 ±0 .0 2 84 .7 8± 1. 53 10 .0 5± 0. 06 4. 92 ±0 .0 7 2. 06 ±0 .0 2 38 .8 0± 0. 55 0. 46 ±0 .0 1 Ei re ni s pu nc ta to lin ea tu s 16 .2 2 ±0 .1 5 9. 58 ±0 .0 7 1. 70 ±0 .0 1 12 2. 07 ±1 .7 8 7. 63 ±0 .0 8 4. 52 ±0 .0 3 1. 69 ±0 .0 2 27 .0 6± 0. 31 0. 22 ±0 .0 1 Ta bl e 3. ( co nt in ue d) . (c on tin ue d) . 189Blood cell morphology of Turkish herpetofauna Sp ec ie s Er yt hr oc yt es N uc le i L (μ m ) W ( μm ) L/ W ES ( μm 2 ) N L (μ m ) N W ( μm ) N L/ N W N S (μ m 2 ) N S/ ES Ei re ni s ro th ii 14 .7 7± 0. 15 8. 73 ±0 .0 6 1. 69 ±0 .0 2 10 1. 24 ±1 .4 4 7. 84 ±0 .1 2 4. 14 ±0 .0 2 1. 90 ±0 .0 3 25 .4 5± 0. 38 0. 25 ±0 .0 1 H em or rh oi s nu m m ife r 15 .6 1± 0. 10 9. 33 ±0 .0 4 1. 68 ±0 .0 1 11 4. 30 ±0 .9 0 6. 92 ±0 .0 8 4. 52 ±0 .0 1 1. 53 ±0 .0 2 24 .5 3± 0. 24 0. 21 ±0 .0 1 C ol ub ri da e H em or rh oi s ra ve rg ie ri 14 .7 6± 0. 16 9. 95 ±0 .1 1 1. 49 ±0 .0 2 11 5. 38 ±1 .9 1 7. 49 ±0 .0 9 4. 91 ±0 .0 4 1. 53 ±0 .0 2 28 .9 0± 0. 50 0. 25 ±0 .0 1 M al po lo n m on sp es su la nu s 15 .2 4± 0. 13 11 .1 6± 0. 09 1. 37 ±0 .0 1 13 3. 60 ±1 .7 7 7. 42 ±0 .0 9 4. 84 ±0 .0 3 1. 54 ±0 .0 2 28 .1 7± 0. 38 0. 21 ±0 .0 2 N at ri x na tr ix 16 .8 7± 0. 18 10 .1 5± 0. 08 1. 67 ±0 .0 2 13 4. 46 ±1 .6 6 7. 95 ±0 .0 7 4. 56 ±0 .0 2 1. 74 ±0 .0 2 28 .4 9± 0. 30 0. 21 ±0 .0 1 N at ri x te ss el la ta 15 .9 8± 0. 21 7. 92 ±0 .0 9 2. 02 ±0 .0 3 99 .6 1± 2. 13 10 .2 1± 0. 09 5. 04 ±0 .0 4 2. 03 ±0 .0 2 40 .4 6± 0. 61 0. 41 ±0 .0 1 Pl at yc ep s co lla ri s 14 .4 0± 0. 12 10 .0 4± 0. 08 1. 44 ±0 .0 1 11 3. 63 ±1 .5 4 7. 29 ±0 .0 7 4. 62 ±0 .0 2 1. 58 ±0 .0 2 26 .4 2± 0. 26 0. 23 ±0 .0 2 Pl at yc ep s na ja du m 15 .4 7± 0. 14 10 .2 3± 0. 13 1. 52 ±0 .0 1 12 4. 50 ±2 .4 1 8. 44 ±0 .2 3 5. 03 ±0 .0 5 1. 69 ±0 .0 5 33 .3 2± 0. 97 0. 27 ±0 .0 1 Pl at yc ep s ve nt ro m ac ul at us 15 .9 4± 0. 17 10 .6 7± 0. 11 1. 50 ±0 .0 2 13 3. 60 ±2 .1 4 6. 91 ±0 .0 6 4. 49 ±0 .0 3 1. 54 ±0 .0 2 24 .3 3± 0. 22 0. 18 ±0 .0 1 R hy nc ho ca la m us m el an oc ep ha lu s 17 .9 6± 0. 20 9. 85 ±0 .0 7 1. 83 ±0 .0 2 13 8. 88 ±1 .9 0 7. 95 ±0 .0 8 4. 47 ±0 .0 2 1. 78 ±0 .0 2 27 .8 8± 0. 28 0. 20 ±0 .0 1 Sp al er os op hi s di ad em a 15 .7 4± 0. 18 9. 52 ±0 .1 3 1. 66 ±0 .0 2 11 8. 10 ±2 .6 9 6. 81 ±0 .1 0 4. 67 ±0 .0 2 1. 46 ±0 .0 2 24 .9 8± 0. 42 0. 21 ±0 .0 2 Te le sc op us fa lla x 18 .3 3± 0. 23 10 .3 3± 0. 10 1. 78 ±0 .0 2 14 8. 80 ±2 .5 7 7. 53 ±0 .0 6 5. 06 ±0 .0 4 1. 49 ±0 .0 2 29 .8 7± 0. 34 0. 20 ±0 .0 2 Te le sc op us n ig ri ce ps 18 .5 5± 0. 20 10 .4 3± 0. 11 1. 78 ±0 .0 2 15 2. 14 ±2 .7 9 7. 96 ±0 .1 0 4. 60 ±0 .0 2 1. 73 ±0 .0 2 28 .7 3± 0. 37 0. 19 ±0 .0 2 Z am en is h oh en ac ke ri 17 .6 6± 0. 24 9. 91 ±0 .0 9 1. 79 ±0 .0 3 13 7. 55 ±2 .5 1 8. 49 ±0 .1 3 4. 44 ±0 .0 2 1. 92 ±0 .0 3 29 .5 3± 0. 41 0. 22 ±0 .0 1 Z am en is lo ng is si m us 12 .7 1± 0. 15 7. 38 ±0 .0 7 1. 72 ±0 .0 2 73 .8 3± 1. 38 6. 61 ±0 .0 8 4. 56 ±0 .0 3 1. 45 ±0 .0 2 23 .6 3± 0. 31 0. 32 ±0 .0 1 V ip er id ae M ac ro vi pe ra le be tin a 17 .2 1± 0. 25 9. 83 ±0 .1 0 1. 75 ±0 .0 2 13 3. 11 ±2 .6 1 6. 68 ±0 .1 2 4. 74 ±0 .0 5 1. 41 ±0 .0 3 24 .8 7± 0. 51 0. 19 ±0 .0 3 M on tiv ip er a al bi zo na 17 .1 6± 0. 26 9. 67 ±0 .1 3 1. 78 ±0 .0 3 13 0. 72 ±3 .3 1 7. 39 ±0 .1 4 4. 36 ±0 .0 5 1. 70 ±0 .0 1 25 .3 2± 0. 62 0. 20 ±0 .0 1 M on tiv ip er a w ag ne ri 17 .6 3± 0. 20 7. 62 ±0 .1 0 2. 32 ±0 .0 3 10 5. 71 ±2 .2 2 10 .6 1± 0. 07 4. 70 ±0 .0 5 2. 27 ±0 .0 2 39 .1 9± 0. 55 0. 38 ±0 .0 1 M on tiv ip er a xa nt hi na 17 .0 8± 0. 16 7. 20 ±0 .1 0 2. 38 ±0 .0 3 96 .7 8± 2. 08 - - - - - V ip er a er iw an en si s 16 .9 8± 0. 17 7. 58 ±0 .0 8 2. 25 ±0 .0 3 10 1. 16 ±1 .6 5 10 .5 8± 0. 06 4. 91 ±0 .0 4 2. 16 ±0 .0 2 40 .7 7± 0. 45 0. 41 ±0 .0 1 El ap id ae W al te ri nn es ia m or ga ni 16 .2 0± 0. 15 10 .1 4± 0. 10 1. 60 ±0 .0 2 12 9. 12 ±2 .0 5 7. 53 ±0 .0 8 4. 82 ±0 .0 4 1. 57 ±0 .0 2 28 .5 2± 0. 41 0. 22 ±0 .0 1 Ta bl e 3. ( co nt in ue d) . 190 H. Arıkan and K. Çiçek ocytoplasmic ratio in M. caspica and the smallest in T. graeca. Nuclei were found more spherical in turtles than amphibians. In the lizard species examined; mean length, width and size of erythrocytes ranged respectively between 12.43 μm-16.85 μm, 7.51 μm-10.21 μm and 73.27 μm2-130.33 μm2; on the other hand, L/W ratio between 1.56-1.99 (Fig. 1F, G, H, I). In this regard, the long- est erythrocytes were observed in Laudakia stellio; the widest and largest in Varanus gri- seus; the shortest, narrowest and smallest in Ophisops elegans (Fig. 1F). And in terms of L/W ratios, the most strongly ellipsoidal cells were determined in Lacerta pamhylica and the least ellipsoidal cells in Anatololacerta danfordi (Table 3). The longest nuclei were found in L. stellio, the widest and largest in Chamaeleo chamaeleon, the shortest and smallest in Trachlepis aurata; the narrowest in Ablepharus chernovi and Trachlepis vittata. Consider- ing NL/NW ratios, the most strongly ellipsoidal nuclei were found in Eumeces schneideri, and the least ellipsoidal in Darevskia uzzelli. The highest nucleocytoplasmic ratio was deter- mined in O. elegans, and the smallest in T. aurata and Chalcides ocellatus (Table 3). In the snake species examined; mean length, width and size of erythrocytes ranged respectively between 14.13 μm-18.55 μm, 7.20 μm-11.16 μm and 84.78 μm2-152.14 μm2; and L/W ratio between 1.37-2.38 (Fig. 1J, K, L). In this regard, the longest and largest erythrocytes were observed in Telescopus nigriceps; the widest in Malpolon monspessula- nus; the shortest in Eirenis eiselti; the narrowest in M. xanthina and the smallest in Eirenis modestus. In terms of L/W ratio; the most strongly ellipsoidal cells were found in M. xan- thina, and the least ellipsoidal in M. monspessulanus (Table 3). Because of the irregular nuclei shapes of erythrocytes in M. xanthina, measurements of nuclei were not given in Table 3. Among the examined species, the longest nuclei were observed in Montivipera wagneri; the widest in Telescopus fallax; the largest in Dolichopis jugularis; the shortest in Macrovipera lebetina; the narrowest in Eirenis rothi, and the smallest in Platyceps ventro- maculatus. In terms of NL/NW ratio; the most strongly ellipsoidal nuclei were determined in M. wagneri; the least ellipsoidal in M. lebetina; the highest nucleocytoplasmic ratio in Vipera eriwanensis, and the smallest in Zamenis longissimus (Table 3). According to the data obtained in the study, there was no correlation between body size and their erythro- cytes size (kendall τ test, r = 0.024, P≤ 0.845). Regarding leucocytes, both small and large lymphocytes were observed as the domi- nant cells in blood smears of all species in herpetofauna. Lymphocytes and monocytes were formed by 80% in leucocytes of the examined species. In small lymphocytes, chromophilic nuclei almost filled the whole cell. Cytoplasm was pushed to a small zone (Fig. 2A). The biggest mean diameter of small lymphocytes was observed in urodeles (14.92 µm), and the smallest (7.79 µm) in lizards (Table 4). Spherical nuclei were more chromophilic in large lymphocytes, and localized in a certain cell zone. Cytoplasm covered larger area than small lymphocytes and was stained a pale blue, and nuclei was stained a purplish blue with Wright’s stain (Fig. 2B). The biggest mean diameter in large lymphocytes was observed in urodeles (20.73 µm), and the smallest (11.63 µm) in lizards (Table 4). Monocytes were similar to large lymphocytes; however, could easily be differentiated by kidney shaped nuclei. Cytoplasm was stained a light gray, and the nuclei was stained a dark purplish blue with Wright’s stain (Fig. 2C, D). The biggest mean diameter in mono- cytes was observed in urodeles (21.00 µm), and the smallest (12.20 µm) in turtles (Table 4). No monocyte was observed in M. lebetina (a snake species). 191Blood cell morphology of Turkish herpetofauna The biggest mean diameter in heterophils of granulocytes, spherical cells, was observed in urodeles ((22.78 µm), and the smallest (11.49 µm) in turtles (Table 4). Their cytoplasms were stained a light blue, and the nuclei, consisting of 2-3 lobes, was stained a red to brown with Wright’s stain (Fig. 2C, E, F). The granules are eosinophilic, elongated, or spindle shaped and could be numerous. Cytoplasms of eosinophils were stained a light yellowish color with Wright’s stain. Since nucleus was masked by the large and bright red granules in cytoplasm, its shape couldn’t be fully distinguished (Fig. 2G, H). The biggest mean diameter in eosinophils was observed in urodeles (21.13 µm), and the smallest in lizards (Table 4). No eosinophil was observed in W. morgani. Fig. 2. Photomicrographs of leucocytes and thrombocytes of some species belong to Turkish Her- petofauna. A: Small lymphocyte (L. trilineata); B: Large lymphocyte (Z. hohenackeri); C: Monocyte and Heterophile (O. elegans); D: Monocyte (P. najadum), E: Heterophile (N. strauchi), F: Heterophile (E. modestus), G: Eosinophile (P. najadum), H: Eosinophile (P. caralitanus), I: Basophile (A. cappa- docica), J: Basophile (S. diadema), K: A group of thrombocytes (O. elegans), L: A group of thrombo- cytes (P. najadum). Horizontal bar: 20 μm. 192 H. Arıkan and K. Çiçek The biggest mean diameter determined in basophils which was smaller than other granulocytic cells was observed in urodeles (19.41 µm), and the smallest (9.99 µm) in lizards (Table 4). Their cytoplasms were filled with black granules, and the nucleus was masked by granules just like in the eosinophils (Fig. 2I, J). No basophile was observed in W. morgani. Thrombocytes were observed as spindle shaped in some species (Fig. 2L), and as nearly spheroidal in others (Fig. 2K). Chromophilic nuclei were found to fill nearly the whole cell. The longest and largest thrombocytes were observed in urodeles (TL = 24.13 µm, TW = 12.88 µm), and the shortest and narrowest in lizards (TL = 7.13 µm, TW = 5.02 µm, Table 4). DISCUSSION AND CONCLUSIONS As stated in literature (Wintrobe, 1933; Foxon, 1964; Hartman and Lessler, 1964; Kuramoto, 1981; Claver and Quaglia, 2009), findings of the study clearly demonstrated that urodeles had the biggest blood cells (erythrocyte, leucocyte, thrombocyte) among the amphibians and reptiles of herpetofauna, and blood smears displayed considerable inter- specific and even intraspecific variations in terms of cell sizes (Fig. 1, 2; Table 2, 3). No important difference was observed between both the erythrocyte and nucleus sizes of urodele and anuran species of the examined amphibians. However, it is impossible to attribute these differences to the correlation regarding body weight and size, defined by Vernberg (1955). More probably, these differences were derived from various environmen- tal conditions (e.g. temperature, air pressure) (Ruiz et al. 1983, 1989) and/or various activ- ity levels (e.g. healthy, breeding, hibernating, foraging, and daily activity) (e.g. Wojtaszek et al., 1997; Campbell, 2004; Allander and Fry, 2008, Sykes and Klaphake, 2008), for erythrocytes were found larger in aquatic species than terrestrials, and smaller in more active species. This view is compatible with the conclusions of Haden (1940), Altman and Dittmer (1961), Harris (1963), Atatür et al. (1998, 1999) and Gül and Tok (2009). Table 4. Leucocytes and thrombocytes size (± with their standard errors) in the peripheral bloods of Turkish amphibians and reptiles [TL: Thrombocyte length, TW: Thrombocyte width]. Lymphocyte (small) (μm) Lymphocyte (Large) (μm) Monocyte (μm) Heterophile (μm) Eosinophil (μm) Basophil (μm) Thrombocytes TL (μm) TW (μm) Amphibia Urodela 14.92±0.20 20.73±0.48 21.0±0.18 22.78±0.14 21.13±0.18 19.41±0.67 24.13±0.64 12.88±0.23 Anura 9.68±0.14 12.46±0.23 14.75±0.34 13.63±0.19 11.61±0.16 11.77±0.20 8.82±0.18 5.93±0.12 Reptilia Testudines 8.52±0.16 11.91±0.18 12.20±0.14 11.49±0.71 11.80±0.32 10.73±0.10 13.68±0.35 6.27±0.25 Squamata Sauria 7.79±0.13 11.63±0.25 13.52±0.28 12.39±0.21 10.47±0.14 9.99±0.10 7.13±0.16 5.02±0.06 Ophidia 7.99±0.11 12.48±0.20 12.85±0.12 11.87±0.31 11.00±0.12 10.52±0.08 10.17±0.42 5.95±0.13 193Blood cell morphology of Turkish herpetofauna L/W ratio ranged between 1.63-1.80 in urodeles, and 1.38-1.69 in anurans (Table 2); consequently, erythrocyte shape was more ellipsoidal in urodeles than anurans. NL/NW ratio ranged between 1.55-1.69 in urodeles, and 1.57-2.35 in anurans (Table 2); that is, contrary to the situation in L/W, anurans were found to have more ellipsoidal nucleus than urodeles, which was compatible with the findings of Kuramoto (1981). Nucleocyto- plasmic ratio ranged between 0.22-0.34 in urodeles, and 0.10-0.16 in anurans (Table 2); that is, anurans had wider cytoplasmic surface area than urodeles in terms of the nuclear surface area in erythrocytes. Therefore, erythrocytes in anurans were more convenient for gas exchange than urodeles. Wintrobe (1933) stated that erythrocyte size reflected the place of a species in the evolutionary scale where the lower vertebrate and the species which were unsuccessful from evolutionary aspect had larger and nucleated erythrocytes; on the other hand, the higher vertebrates had small and enucleated erythrocytes. From this respect, reptiles are regarded as intermediate between amphibians and birds (Szarski and Czopek, 1966; Szar- ski, 1968). Results of the study indicate that erythrocyte size reflected the place of a spe- cies in the evolutionary scale in higher taxa. Different researchers (Hartman and Lessler, 1964; Szarski and Czopek, 1966; Saint Girons and Saint Girons, 1969; Saint Girons, 1970; Arıkan et al., 2004; Frye, 1991; Mader, 2000; Campbell, 2004; Strik et al., 2007; Sykes and Klaphake, 2008; Arıkan et al., 2009a, b; Claver and Quaglia, 2009) reported that reptiles constitute a heterogeneous group among vertebrates in terms of their blood cell morphology, and demonstrated considerable varia- tions among orders, even within the same family members. Among reptiles, the largest erythrocytes were observed in Sphenodon punctatus, in turtles and crocodiles; and the smallest in lacertid lizards (Hartman and Lessler, 1964; Saint Girons and Saint Girons, 1969; Saint Girons, 1970; Sevinç et al., 2000). Among the turtle species examined, aquatic ones had larger erythrocytes and nuclei than terrestrial T. graeca (Table 3). Aquatic species had more ellipsoidal erythrocytes than T. graeca regarding L/W ratio; however, T. graeca had more ellipsoidal nuclei than aquat- ic species regarding NL/NW ratio (Table 3). This confirms the findings of Uğurtaş et al. (2003). Nucleocytoplasmic ratio was found smaller in T. graeca than aquatic species (Table 3). Consequently, it can be concluded that T. graeca had more convenient erythrocytes for gas exchange than aquatic ones. Among the 30 lizard species examined, the largest erythrocytes were observed in V. griseus and the smallest in O. elegans. Erythrocyte size demonstrated great variations among the families, and in some cases even within the species of the same family, which we believe were caused by different activity levels (e.g., healthy, breeding, hibernating, for- aging, and other daily activities). Regarding L/W ratio, the most ellipsoidal erythrocytes were observed in L. pamhylica, and the least or nearly spheroidal ones in A. danfordi. Regarding NL/NW ratio, scincid lizards had more ellipsoidal nucleus than others (Table 3). Generally, there was a positive correlation between erythrocyte and nucleus sizes in lizards. Nucleocytoplasmic ratio ranged between 0.12-0.15 in Scincidae family, and 0.19- 0.27 in others (Table 3). In this regard, we could deduce that Scincidae had more conven- ient erythrocytes for gas exchange than other lizards. Saint Girons and Saint Girons (1969) reported that except for Typhlops vermicularis with their relatively small erythrocytes and large nuclei, the snakes formed a homoge- 194 H. Arıkan and K. Çiçek nous group regarding their erythrocyte sizes. However, in this study, great variations were found among families, and even within the same family members regarding their eryth- rocyte sizes. Among the 34 species examined, the largest erythrocytes were observed in T. nigriceps, and the smallest in Z. longissimus. Regarding L/W ratio, the most ellipsoidal erythrocytes were in M. xanthina, and the least ellipsoidal or nearly spheroidal ones in M. monspessulanus. Results of the study indicated that small sized species (e.g., T. vermicula- ris’s mean size is 25 cm) don’t have smaller erythrocytes than the biger ones (e.g., Z. long- issimus’s mean size is 150 cm). Some researchers (Gulliver, 1875; Saint Girons and Saint Girons, 1969; Arıkan et al., 2004) reported the presence of somewhat irregular nuclei in the erythrocytes of viperid and elapid species. Similar results were found especially in M. xanthina. Regarding NL/ NW ratio, the most ellipsoidal nuclei were observed in M. wagneri, and the least ellipsoi- dal in M. lebetina. Contrary to lizards, there was no correlation between the erythrocyte and nuclei sizes in snakes. Regarding nucleocytoplasmic ratio, snakes formed a heteroge- neous group, and this ratio ranged between 0.19 and 0.46. Lymphocytes are generally dominant leucocytes in amphibians and reptiles (Frye, 1991; Mader, 2000; Campbell, 2004; Strik et al., 2007; Allander and Fry, 2008; Sykes and Klapha- ke, 2008). Saint Girons (1970) and Arıkan et al. (2004, 2009a) reported that small and large lymphocytes were the dominant cells in blood smears of different reptile species, and the nuclei were not easily be distinguished, for they were masked by dense granulations in the cytoplasms of both eosinophils and basophils. In this study investigating both amphibian and reptile species of herpetofauna; the largest leucocytes were found in urodeles (Table 4); small and large lymphocytes were the dominant cells in the blood smears; and the shapes of the nuclei were not distinguished because of the dense granulations in the cytoplasms of both the eosinophils and basophils, which were all compatible with the literature (e.g., Claver and Quaglia, 2009). Though monocytes, heterophils, and eosinophils were present in amphibians and reptiles (Allander and Fry, 2008; Sykes and Klaphake, 2008), Cannon et al. (1996) reported that the heterophils were not observed in Cyrtopodion scabrum. Besides, eosinophils were observed in Crocodilia and Chelonia, but their existence in Squamata is controversial (Claver and Quaglia 2009). Even inside a genus of snakes, eosinophils were found in some species and not found in some others (Alleman et al., 1992; Troiano et al., 1997). Number and kind of leucocytes could be change environmental and physiological activity (Allander and Fry, 2008; Sykes and Klaphake, 2008). However, blood cells of reptiles still completely unknown (e.g., Frye, 1991; Mader, 2000; Campbell, 2004; Strik et al., 2007), five types of leucocytes are observed (lymphocyte, monocyte, heterophile [neutrophile], eosinophile and basophile) (Sykes and Klaphake, 2008). Thrombocytes were defined by some researchers as spindle shaped cells with central- ly localized extremely chromophilic nuclei (Saint Girons, 1970; Canfield and Shea, 1988; Arıkan et al., 2004, 2009a; Allander and Fry, 2008; Sykes and Klaphake, 2008). Spindle- shaped thrombocytes were observed in some species of herpetofauna, and nearly spheroi- dal ones were found in some other species. In this study, the largest thrombocytes were observed in amphibians and the smallest in lizards (Table 4). 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