Comparative serological and hematological studies of three anatolian mountain frogs

Mountain frogs belonging to the subgenus Rana are distributed at the mountainous regions ofAnatolia, Caucasus, Northern Iran and probably of Northern Iraq. Works on the taxonomical states andthe geographical ranges of the Anatolian mountain frogs go back to the year 1885. The first describedspecies was Rana mocrocnemis from Uludağ by Boulunger (1885). An other species, R. camerani,was first described from Tabizhuri Lake and Achalki in Caucasus by Boulenger (1886), later was alsoreported from Erciyes Mt. İn Anatolia by Werner (1902). A third species, R. holtzi, was described fromthe Maden Lake in the Taurus Range, again by Werner (1898).Following the initial descriptions, the taxonomical states of the three Anatolian mountain frogswere evaluated variously by different authors. Boulunger (1898) was of the opinion that R.holtzi was asynonym of R.camerani, a view initially accepted by Werner (1902), but later, the latter author revertedto his former belief that R. holtzi was a distinct species. Lantz and Cyren (1913) and Bodenheimer(1944) asserted that R.camerani was identical with R. macrocnemis: According to Delwig (1928),these were the geographical races of the same species; while Terent?ev and Chernov (1949)maintained that R. macrocnemis and R.camerani were separate and distinct species.Baran (1969), who worked on the morphological characteristics of large samples of Anatolianmountain frogs, and according to Özeti (1970) and Kaya (1996), who examined the same materialosteologically, three distinct species live in Turkey. Baran and Atatür (1986) described a new racefrom Akdağ-Tavas as R.m. tavasensis. More recently Tarkhnishvili and Gokhelashvili (1999) andTarkhnishvili et al., (1999) pointed out the presence of two geographical sobspecies within theR.macrocnemis group: R.m. macrocnemis and R. m. camerani. Arıkan et al., (2001) stressed that nosignificant morhological differences were present among the populations from Çamlıyayla, Bolkar Mts.and Aladağ Mt. at the middle Taurus Range only some pattern and coloration differences wereevident.The samples used in this study were collected from various localities of Anatolia, between theyears 2002-2004.Freshly caught specimens were analiyzed according to their pattern and colorationcharacteristics, color dias were also photographed. Measurements and ratios were taken according toTerent?ev and Chernov (!949) and Baran ( 1969).This study is on the establishment of erythrocyte and nuclei sizes in three mountain frogs fromAnatolian, by means of blood smears stained with Wright?s stain. Largest erythrocytes were observedin the smears of R.macarocnemis (Sultan Mts.), while the smallest in R.macrocnemis (Kaçkar Mts.)Among the studied species, the nuclei and erythrocyte sizes were found significantly correlated.Body proportions of the seven populations were compared using statistical analysis.Sexually mature specimens were used in this study. Among the investigated characteristics, nosignificant differences were observed between the sexes, so, data obtained from the males and thefemales were pooled, than evaluated.Blood samples were taken with heparinized hematocritcapillaries, utilizing cardiac (ventricle) puncture, centrifuged 600 g?de for five minutes and the obtainedblood- serum were kept at -20 oC until electrophoretic separation. Polyacrilamide disc electrophoreseswere conducted within a Canalco Model 1200 electrophoresis apparatus, according to the method of1Davis (1964), slightly modified by Arıkan (1983). A 5 microlitre blood serum sample was used for eachelectrophoretic separation. The separating gels were than stained with 0.5 % Amidoblack in 7% aceticacid. Passive destaining was done within 7% aceticacid baths.While three species of mountain frogs (R. macrocnemis, R. camerani and R. holtzi) have beenreported from Anatolia in the relevant literature, a consensus has not been reached yet between theauthors. Especially the taxonomical states of R. macrocnemis and R.camerani are not clear. Lantz andCyren (1913) and Bodenheimer (1944) accepted these species as identical to each other; according tothe Delwig (1928) they are races the same species, while Terent?ev and Chernov (1965), Baran(1969), Özeti (1970), Kaya (1996) and Başoğlu et al. (1994) were of the were of the opinion that theseare distinct species. Baran and Atatür (1986) named the Akdağ population as a new subspecies,R.m.tavasensis. Ischenko (1978, 1987), who works on Caucasian brown frogs, claimed that R.camerani is neither a distinct species nor a subspecies, but in the related region only R.macrocnemiswas present as a greatly variable monotypical species.Tarknishviliv and Gokhelashvili (1999)mentioned the presence of two geographical races as R.m.macrocnemis and R.m.camerani in theCaucasus region, and between these two races reported some differences concerning the skinstructure, condition of the vertebral stripe and the size of the metatarsel tubercle; i.e. in R.m.cameranithe skin was rough dorsal maculations were distinct, a vertebral stripe was present and the metarseltubercle was bigger while in R.m.macrocnemis the skin was smooth, dorsal maculations were faded,vertebral stripe was absent and the metarsaltubercle was smaller. According to Tarknishvili etal.(1999) two geographical taxa were present in the Caucasus, R.m.macrocnemis and R.m.camerani,which were discernible with eight morphometrical characters.Our findings indicate that the specimens from Karagöl population (Bolkar Mts.) had hadsmooth skins, their variously sized and shaped dorsal maculations were encircled with lighter coloredrims. None of the investigated specimens had avertebral stripe. These findings are completely inaccord with those of Werner (1898), the original describer of R.holtzi. this is quite natural, since thispopulation is composed of topotypical specimens. On the other hand, the general description ofTarkhnishvili and Gokhelashvili (1999) on R.m.macrocnemis from Caucasus includes some similaritiesto the description of our R. holtzi material. We believe, that material from Caucasus should bereconsidered taxonomically, their characteristics seem to be closer to R.holtzi than those of thenominate race. No significant differences were observed between the population from Uludağ, Akdağ,Bolkar Mts, Erciyes Mt., Sultan Mts., Kaçkar Mts, Karçal Mt; from the viewpoints of morphometricalmeasurements and ratios, skin structure, pattern and coloration characteristics of the dorsum,presence of vertebral stripe and blood serum proteins.The results of our morphometrical and serological analyses indicate that only two distinctmountain frog species, R.holtzi and R.macrocnemis inhabit the three known localities of Anatolia; i.epreviously reported R.camerani (of Mt Erciyes) could be accepted as a synonym of R.macrocnemis.2