Hüseyin Arikan

Determination of in vivo toxicity and in vitro cytotoxicity of venom from the Cypriot blunt-nosed viper Macrovipera lebetina lebetina and antivenom production

The venomous Levantine viper, Macrovipera lebetina lebetina is endemic to Cyprus. The objective of this study was to investigate in vitro cytotoxicity, in vivo lethality, and antivenom production followed by a re-immunization schedule in mice against Macrovipera lebetina lebetina venom. The LD50 value was estimated as 7.58 mg/kg within 24 hours by different venom doses administrated intraperitoneally in mice. Freunds complete and incomplete adjuvants were used for first and second immunization of mice in antivenom production. A cell-based assay was performed to determine the effects of Macrovipera lebetina lebetina venom and antivenom neutralizing potency on L929 cell viability. The snake venom toxicity and cytotoxicity were examined and comparison of results showed good correlation, the LD50 value was tenfold higher than the IC50 value. The IC50 value was 0.62 ± 0.18 mL after 48 hours treatment while the calculated value was 1.62 ± 0.25 mL for the culture media totally refreshed after two hours treatment with venom. The in vitro efficacy of antivenom against Macrovipera lebetina lebetina venom was found to be low. This is the first report that describes the in vivo and in vitro toxic effects of Macrovipera lebetina lebetina venom and antivenom production against this species.

Histochemical and Biometric Study of the Gastrointestinal System of Hyla Orientalis (Bedriaga, 1890) (Anura, Hylidae)

This study was carried out to assess the localization of hyaluronic acid (HA) and the distribution of glycoproteins in the gastrointestinal system of adult Hyla orientalis. Histochemical analysis of the gastrointestinal system in H. orientalis showed that mucous content included glycogene and/or oxidable dioles [periodic acid/Schiff (PAS)+], neutral or acid-rich (PAS/AB pH 2.5+), sialic acid residues (KOH/PAS+) and acid sulphate [Aldehyde fuchsin (AF)+] glycoproteins. However the mucus content was not the same in stomach, small and large intestine. The mucus content of stomach included only glycogene and/or oxidable dioles and sialic acid residues. Besides these histochemical methods, the localization of HA was detected using biotinylated hyaluronic acid binding protein labeled with streptavidin-fluorescein isothiocyanate (FITC). In the extracellular matrix of the submucosa, the reaction for HA was evident. Since HA was located in submucosa beneath the epithelial layer of gastrointestinal system, it has a significant role in hydric balance, and essential to provide the gastrointestinal system integrity and functionality. According to biometric results, there were statistical differences between small and large intestine in terms of the amount of material stained positive with PAS/AB, PAS, KOH/PAS and AF/AB. Additionally, number of goblet cells in the small and large intestine was significantly different.

Comparative morphological and serological studies of three Anatolian Mountain frogs, Rana macrocnemis , R. camerani and R. holtzi (Anura, Ranidae)

Three currently recognized species of Anatolian mountain frogs were analyzed for species boundaries. Meristic and proportional measurement data of three populations, including two topotypical ones, were evaluated using both univariate and discriminant analyses, and blood-serum samples were analyzed electrophoretically with polyacrylamide-disc electrophoresis. The results indicate that only two distinct mountain frog species inhabit Anatolia. We recommend that R. camerani from Mt Erciyes should be recognized as a synonym of R. macrocnemis.

A study on the blood cells of the Caucasus Frog, Pelodytes caucasicus

Abstract This study gives detailed information on the size and number of the various blood cells from a sample of the Caucasus Frog, Pelodytes caucasicus, based on 10 ♂ and 10 ♀ from Uzungöl (Trabzon province). P. caucasicus does not show sexual dimorphism in the number and size of blood cells; the erythrocytes of P. caucasicus are smaller than those of the other Turkish anurans, but their number is higher.

A study on the blood cells of the fire-bellied toad, Bombina bombina L. (Anura: Bombinatoridae)

The aim of this study was to determine the blood cells parameters of Anatolian and Thracian Bombina bombina populations. The size and counts of various blood cells were analyzed in 29 mature Bombina bombina specimens belonging to two populations (Durusu-Istanbul and Doseme Batakligi–Adapazari) in northwest Anatolia. The mean erythrocyte count was determined as 251.429/mm3, that of leucocytes was 2.114/mm3, and that of thrombocytes was 12.857/mm3. There was no significant sexual variation in blood cell counts of our B. bombina sample which was collected during their breeding season. The leucocyte differential counts and staining properties of blood cells were also determined. Interspecific comparisons indicate that northwest Anatolian B. bombina samples share general features with other anurans but differ from other populations of the same species in blood cell parameters. The lower number of erythrocyte and their relatively large surface area of our samples suggest a lower level of metabolism and relatively higher oxygen uptake. We suggest that this structure is probably not only due to adaptations to different geographic regions and altitude but also a combination of recent habitat reductions due to human actions and agricultural contaminants may account for the blood cell parameters as well as other genetic and morphologic patterns of these isolated and small populations of this species.

Electrophoretic Comparison of Blood-serum Proteins of Apathya cappadocica (Sauria, Lacertidae) Subspecies from Anatolia

Blood-serum proteins of the known subspecies of Apathya cappadocica (Werner, 1902) were studied comparatively by polyacrylamide disc gel electrophoresis. In order to obtain useful biochemical data for classification, differences between the electrophoreograms of the samples included in the morphologically different subspe - cies were distinguished qualitatively and quantitatively. These comparisons indicated that electrophoretic results supported morphological discrimination of the known subspecies of A. cappadocica.

Investigation of dorsal/ventral skin and the parotoid region of Lyciasalamandra billae and Lyciasalamandra luschani basoglui (Urodela: Salamandridae)

In the present study, the parotoid region, dorsal and ventral integuments of Lyciasalamandra billae and Lyciasalamandra luschani basoglui were investigated in terms of localization of hyaluronic acid (HA) and histochemical characteristics. HA immunoreactivity was carried out using biotinylated hyaluronic acid binding protein (B-HABP) labelled with streptavidin-fluorescein isothiocyanate (FITC). HA was mainly localized in the stratum spongiosum of L. billae and L. luschani basoglui for water homeostasis and skin functionality. Light microscopic observations revealed that the dorsal and ventral integuments of L. billae and L. luschani basoglui exhibited basic morphological characteristics of other amphibians: the epidermis was composed of a stratified squamous epithelium and the dermis subdivided into stratum spongiosum and stratum compactum. Two different types of dermal glands (mucous and granular glands) were identified in the spongious dermis of the dorsal and ventral integuments whereas in the parotoid region, three different types of glands (mucous, granular and parotoid glands) were examined.

Morphological and serological comparison of Mediterranean Spur-Thighed Tortoises, Testudo graeca, from the Aegean region and southeastern Turkey

Abstract We examined a total of 29 specimens of the Mediterranean Spur-thighed Tortoise, Testudo graeca, from two localities in the Aegean region and southeastern Turkey: 7♂ and 9♀ from İzmir and 4♂ and 9♀ from Adıyaman. An analysis of variance based on 31 morphometric ratios revealed significant differences (p<0.05) in the ratios of straight carapace length/femoral suture length, straight carapace length/plastron width 2, plastron length/plastron width 2, carapace width/carapace height, carapace width/gular suture length, carapace width/femoral suture length, nuchal length/nuchal width, and carapace width/plastron width 2. Both populations can be characterized by those parameters. The colour and pattern were inconsistent in both populations, but black is generally dominant in the Aegean population, whereas the prevailing colour in the southeastern population is yellow. A quantitative and qualitative comparison of the electrophoretic pattern (electropherograms) of the blood-serum proteins of both populations revealed no differences.

Genetic Differentiation Among Peripheral Populations of Bombina bombina from Thrace and Anatolia: An Allozyme Analysis

Genetic structures of Bombina bombina populations, located as peripheral isolates in Turkish Thrace and northwestern Anatolia, were analyzed by polyacrylamide gel electrophoresis using 20 allozyme loci, to investigate the populations’ current genetic variation and possible colonization history. Significant genetic variability was detected in most of the loci and all populations. Allozyme pairwise FST matrices and distribution of allele frequencies indicate their very close genetic relationships and relatively recent formation. Mean genetic distance values between Thracian and Anatolian populations indicate a Middle or Upper Pleistocene lineage separation before the formation of the Bosporus as an isolating geographic barrier. All the samples show substantial heterozygosity excess, and there was statistically significant evidence of recent bottlenecks. The extent and patterns of genetic divergence indicate that the Anatolian and Thracian populations have probably experienced bottlenecks, and incipient speciation may have occurred in Anatolian populations of B. bombina.

Serological characterization and confirmation of the taxonomic status of Montivipera albizona (Serpentes, Viperidae) with an additional new locality record and some phylogenetical comments

Th is is the fi rst record of the presence of Montivipera albizona (Nilson, Andren & Flardh 1990), in Kahramanmaras province, Mediterranean Region of Turkey. Here, one young male specimen was collected and is described. Th e present record of M. albizona extends its known distribution (Kulmac Mountain Range, Sivas) some 250 km to the south-west, where the Anatolian Diagonal exhibits a bifurcation. Our data based on the electropheoretic analysis of blood-sera, indicate that the M. xanthina populations from the western Anatolia and M. albizona distributed along the Anatolian Diagonal show signifi cant diff erences, qualitatively and quantitatively. Accordingly, it is concluded that M. albizona should not be included within the polymorphic species M. xanthina which lives in western Anatolia and therefore, it should be accepted as a valid species. Moreover, the actual distributions of the related taxa in Anatolia and the geographic structure of the Anatolian Diagonal were discussed.