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): Vergilov Vladislav S., Zlatkov Boyan, Tzankov Nikolay D. Artikel/Article: Hemipenial differentiation in the closely related congeners Ablepharus kitaibelii (BIBRON & BORY DE SAINT-VINCENT, 1833), and Ablepharus budaki GÖCMEN, KUMLUTAS & TOSUNOGLU, 1996 (Squamata: Sauria: Scincidae) 39-48 Vergilov_et_al_ablepharus_hemipenis:herPeToZoa.qxd 07.08.2017 18:15 Seite 1 herPeToZoa 30 (1/2): 39 - 48 39 Wien, 30. Juli 2017 hemipenial differentiation in the closely related congeners Ablepharus kitaibelii (B iBroN & B ory De SaiNT -V iNceNT , 1833), and Ablepharus budaki göçmeN , k umluTaş & T oSuNoğlu , 1996 (Squamata: Sauria: Scincidae) unterschiede in der hemipenismorphologie der beiden nahe verwandten arten Ablepharus kitaibelii (B iBroN & B ory De ST-V iNceNT , 1833) und A. budaki göçmeN , k umluTaş & T oSuNoğlu , 1996 (Squamata: Sauria: Scincidae) VlaDiSlaV S. V ergiloV & B oyaN P. Z laTkoV & N ikolay D. T ZaNkoV †) kurZFaSSuNg Die hemipenes zweier nahe verwandter Ablepharus arten, A. kitaibelii (BiBroN & B ory De SaiNT -V iNceNT , 1833 ) und A. budaki göçmeN , k umluTa ş & T oSuNoglu , 1996 werden erstmalig beschrieben. Diese arten haben ähnliches aussehen und wurden nur durch wenige externe und sehr variable merkmale unterschieden. in Über ein - stimmung zu einer früheren genetischen Studie zeigten die männlichen geschlechtsorgane jedoch deutliche morpho - logische unterschiede. einige merkmalsausprägungen der hemipenes weisen Ähnlichkeiten mit denen der Skink- Sub familie lygosominae auf. Darüberhinaus wird eine bisher unbeschriebene hemipenisstruktur als ‘asulcal pro - trusion’ beschrieben. es wurden ausschließlich epithelial verhornte, unverkalkte mikrostrukturen beob achtet. eine neuartige methode des ausstülpens kleiner, fragiler hemipenes von eidechsen wird vorgeschlagen. aBSTracT The hemipenes of two closely related Ablepharus species, A. kitaibelii (BiBroN & B ory De SaiNT -V iNceNT , 1833), and A. budaki göçmeN , k umluTaş & T oSuNoglu , 1996, are described for the first time. These species are of similar general appearance and differ by few external and variable characters only. The genital morphology, however, revealed profound differences, which is in concert with a previous genetic study. Some hemipenial char - acters resemble those found in lygosomine skinks. moreover, a previously undescribed hemipenial structure was found: the asulcal protrusion. only epithelial non-calcified keratinous microstructures were observed. an inno - vative technique for eversion of small and fragile lizards’ hemipenes is proposed. key WorDS reptilia: Squamata: Sauria: Scincidae; Ablepharus kitaibelii , Ablepharus budaki , hemipenis, bulbous lobes, asulcal protrusion, morphology, microstructures, eversion iNTroDucTioN The genus Ablepharus FiTZiNger in lindbergi WeTTSTeiN , 1960; A. pannonicus eVerSmaNN , 1823, currently comprises ten (FiTZiNger , 1824), and A. rueppellii (gray , species [ Ablepharus bivittatus méNéTrieS , 1839)] confined mainly to southwestern and 1832; A. budaki göçmeN , k umluTaş & southern asia ( SiNDaco & J eremčeNko ToSuNoğlu , 1996; A. chernovi DareVSky , 2008). a single species, A. kitaibelii , is 1953; A. darvazi JeriomčeNko & P aNFiloV , present in europe. Taxonomic revisions 1990; A. deserti STrauch , 1868; A. gray - (STePáNek 1937; m erTeNS 1952; F uhN anus (STolicZka , 1872), A. kitaibelii (BiB- 1970; e remčeNko & S čerBak 1986; g öç- roN & B ory De SaiNT -V iNceNT , 1833), A. meN et al. 1996), resulted in descriptions of ______________________________________ †) The authors Vladislav S. VergiloV and Boyan P. ZlaTkoV dedicate this paper to their good friend and esteemed colleague Nikolay D. TZaNkoV , who recently passed away in a tragic car accident. Vergilov_et_al_ablepharus_hemipenis:herPeToZoa.qxd 07.08.2017 18:15 Seite 2 40 V. S. V ergiloV & B. P. Z laTkoV & N. D. T ZaNkoV several subspecies: A. k. fabichi STèPàNek , Ver & B öhme 1986), lacertidae ( Böhme 1937; A. k. fitzingeri merTeNS , 1952; A. k. 1971; a rNolD 1983, 1986a, 1986b), gymn - stepaneki FuhN , 1970, and A. k. budaki . ophthalmidae ( uZZel 1969, 1973; m yerS et Shortly after its description, the status of the al. 2009; NuNeS et al. 2012, 2014), Dac - latter taxon was raised to full species by tyloidae ( köhler 2010; k öhler & k reuTZ SchmiDTler (1997). he also introduced a 1999; k öhler & S uNyer 2008; k öhler & new subspecies A. b. anatolicus from the VeSely 2010; k öhler et al. 2007, 2010, anatolian mainland. Based on molecular 2012). generally, the members of the Scin - data, PoulakakiS et al. (2013) demonstrat - cidae family have been studied sporadically, ed that A. budaki belongs to a well-support - with few works concerning this topic ( Böh- ed clade that is further subdivided into three me 1988; B öhme et al. 2000; greer & B iS- monophyletic lineages. Diagnostic features WaS 2004; Z iegler & B öhme 2004; h orNer and arguments for raising A. budaki to 2007). in Ablepharus this feature received species level included external morphologi - very little attention. up to this date, a single cal traits ( göçmeN et al. 1996; SchmiDTler study dealing with this matter schematically 1997) and substantial genetic divergence presents the hemipenial morphology of A. from A. kitaibelii (Pou lakakiS et al. 2005, darvazi , A. deserti and A. pannonicus 2013). (eremčeNko & P aNFiloV 1990). The hemipenial morphology has The purpose of this study is to proven its significance as an important trait describe the hemipenial morphology of two for interspecific taxonomic differentiation closely related representatives of the genus of congeners. This method has been applied Ablepharus and to propose an alternative for other small representatives of various technique for hemipenial preparation appli - lizard families, e.g., chamaeleonidae ( kla- cable to small lizard species. maTerialS aND meThoDS materials.– Ablepharus kitaibelii , The specimens were dissected and three adult males from Bulgaria: Silistra both hemipenes together with the retractor region, near v. Sratsimir, No. NmNhS194; muscles and connective tissue were re - Southern Pirin mts., near v. Belyovo, No. moved carefully with small scissors, forceps NmNhS45; Strandzha mts., near v. Bele - and scalpel, and then submerged into 5 % vren, No. NmNhS170. Ablepharus budaki , ethanol solution. This ethanol concentra - two adult males from Turkey, hatay Pro - tion decreases the surface tension without vince, near v. ylanlı, NmNhS400-401. all hardening the soft tissues. Then the con - specimens are preserved in ethanol in the nective tissue and most of the retractor mus - collection of the National museum of Nat - cle were extirpated. after that, the hemi - ural history, Bulgarian academy of Scien - penes were everted using fine forceps and ces (NmNhS) in Sofia, Bulgaria. syringe with a thin metal needle. complete methods.– early proposed tech - eversion was achieved after injecting 5 % niques for hemipenial preparation (e.g., or- ethanol into partially everted hemipenes. TeNBurger 1923; B öhme 1971, 1988; Processing with potassium hydroxide maN ZaNi & a Be 1988; P eSaNTeS 1994 etc.) (koh) solution used for various larger rep - appeared to be inapplicable to the studied tiles ( PeSaNTeS 1994; Z iegler & B öhme objects because of their small size and 1997; Z aher , 1999; h arVey et al. 2012) fragility. as small skinks possess delicate and some smaller species ( NuNeS et al. hemipenes the authors used techniques 2014) proved to be inapplicable, because developed for everting the endophallus of the tissues became too soft and fragile, and large moths ( harDWick 1950; D uFay 1978; consequent eversion was impossible. The FiBiger et al. 1984) with some modifica - everted hemipenes were stained in 0,5 % tions. The method is based on manual ever - alizarin red S solution for about 10-15 sec - sion and dehydration of the soft tissue by onds. This stain is used for contrasting of injection with absolute ethanol. hemipenial carbonate structures of lizards Vergilov_et_al_ablepharus_hemipenis:herPeToZoa.qxd 07.08.2017 18:15 Seite 3 hemipenial differentiation in Ablepharus kitaibelii and A. budaki 41 (uZZell 1969, 1973). Both alcohol (70 %) and two of A. budaki were made. For the and water solution of alizarin produced purpose, the hemipenes were slit laterally to equal results. The staining did not provide produce sulcal and asulcal halves, then the expected result because the stain pene - compressed between two slides and dehy - trated in all tissues, hence they were equally drated with absolute ethanol for five min, stained and no contrasted structures were after that removed from the slides and observed under a stereomicroscope. how - cleared with euparal essence for five min ever, this procedure proved to be useful for and mounted permanently on a slide with subsequent microscopic observation. after euparal embedding agent. To test the pres - staining the hemipenes were inflated with ence of carbonate structures, two hemipenes 100 % ethyl alcohol using a syringe with a of A. kitaibelii and one of A. budaki were blunt needle. a continuous flow of alcohol treated with 10 % hNo 3 for a minute (under was maintained for a few seconds to harden direct microscope observation on temporary the hemipenes. To avoid ejecting by pres - slide mounts), then washed with distilled sure, the hemipenes were tied at the