Copyright: © 2013 Rastegar-Pouyani et al. This is an open-access article distributed under the terms of the Creative Commons Attribution–NonCommercial–NoDerivs 3.0 Unported License, which permits unrestricted use for non-commercial and education purposes only provided the original author and source are credited. The oficial publication credit source: Amphibian & Reptile Conservation at: amphibian-reptile-conservation.org Amphibian & Reptile Conservation 6(2): 35–39. New country record and range extension of Eremias suphani Başoğlu & Hellmich, 1968 from Iran 1,5 Eskandar Rastegar-Pouyani, 2Aziz Avci, 3Yusuf Kumlutaş, 3Çetin Ilgaz, and 4Seyyed Saeed Hosseinian Yousefkhani 1 Department of Biology, Faculty of science, Hakim Sabzevari University, Sabzevar, IRAN 2Adnan Menderes University Faculty of Science and Arts, Department of Biology, Aydın-TURKEY 3Dokuz Eylül University, Faculty of Science, Department of Biology, Buca, İzmir-TURKEY 4Young Researchers and Elite Club, Shirvan Branch, Islamic Azad University, Shirvan, IRAN Abstract.—Iran is located in an area that is bordered by several mountains and contains herpetofaunal constituents from adjacent countries such as Pakistan, Iraq, Afghanistan, and Turkey. We surveyed several border provinces of Iran to more completely understand the Iranian herpetofauna. During one survey in the northwestern part of Iran we found a species of the genus Eremias that can be added to Iran’s herpetofauna. Previously, the distribution of Eremias suphani was limited to the Van Lake in Turkey (type locality). We compared Iranian samples with E. suphani from Turkey using a molecular marker (Cytochrome b) and conirmed that this species is also present in Iran. The new record of this species is located on the road from Firoragh to Chaldoran in the west of the province of Azarbaijan, Iran. Key words. Eremias suphani, Iran, Turkey, new record, molecular marker Citation: Rastegar-Pouyani E, Avci A, Kumlutaş Y, Ilgaz C, Hosseinian Yousefkhani SS. 2013. New country record and range extension of Eremias suphani Başoğlu & Hellmich, 1968 from Iran. Amphibian & Reptile Conservation 6(2): 35–39 (e73). Introduction E. suphani), and the presence of a ventrolateral row of well-deined, large, round, white spots, that may be fused to form a longitudinal band (diffuse small longitudinal spots or a diffuse thin line in E. strauchi). The distribution of these two sister species meet in the Dogubayazit Depression in NE Turkey (Bischof and Böhme 1980). In Iran, E. suphani is found on compacted, loamy soil at the Iran-Turkey border. In Turkey, E. suphani also lives on pebbly/sand substrates with sparse vegetation (Franzen and Hecks 1999; Baran et al. 2012). Amphibian and reptile species that have a sympatric distribution with E. suphani include: Bufotes variabilis, Pelophylax ridibundus, Rana macrocnemis, Testudo graeca, Trapelus lessonae, Ophisops elegans, and Lacerta media. Eremias suphani is listed as Least Concern because, although its extent of occurrence is less than 20,000 km2, it is common with large population sizes, no major threats, and it is unlikely to be declining fast enough to qualify for listing in a more threatened category (Kaska et al. 2013). Herein we report a new Iranian country record for E. suphani, conirmed using a molecular marker (Cytochrome b). The herpetofauna of Iran remains poorly documented and in some isolated areas completely unknown (Anderson 1999). Historically, the Iranian herpetofauna has been studied by several foreign herpetologists (Mertens 1957; Anderson 1966, 1999; Leviton et al. 1992; Tuck 1971, 1974) who visited Iran, as well as by Iranian herpetologists (Latii 1991; Balouch and Kami 1995; Kami and Vakilipoure 1996a, 1996b; Firouz 2000; RastegarPouyani et al. 2007). Nonetheless, herein we report the presence of a previously undocumented species from the country. According to the recent published data, 15 species of the genus Eremias exist in Iran (Hosseinian Yousefkhani et al. 2013), to which we add another. Eremias suphani (Suphan Racerunner) was considered to be conined to Turkey, where it was described from Van Lake (Başoğlu and Hellmich 1968), with some additional populations of this species having been recorded from western Turkey (Bischof and Böhme 1980). The species is morphologically similar to Eremias strauchi, its putative sister species, but there are differences in scalation (Bischof and Böhme 1980). Adults of Eremias suphani can be distinguished from E. strauchi by the presence of a double or single row of gular scales separating the scales of the third pair of chin shields (third pair of chin shields in contact in E. strauchi), the presence of enlarged gular scales bordering the third pair of chin shields (no enlarged gulars in Materials and Methods The Iranian samples were collected during ield trips to the northwest of the country in June 2005 on the road Correspondence. 5rastegarpouyani45@gmail.com Amphib. Reptile Conserv. | amphibian-reptile-conservation.org 35 December 2013 | Volume 6 | Number 2 | e73 Rastegar-Pouyani et al. Figure 1. Habitat of Eremias suphani in NW Iran on the road from Firoragh to Chaldoran in Ali Sheykh village (Photo by Eskandar Rastegar-Pouyani, 2005). three specimens of E. strauchi and ive specimens of E. suphani from Turkey where compared with the Iranian samples using MEGA 5.0 software (Table 1). The sequences have been deposited in the GeneBank with accession numbers from KF797802 to KF797813. from Firoragh to Chaldoran, at the village of Alishekh (Fig. 1) (E 44° 34ʹ 78.4ʺ, N 38° 49ʹ 22.1ʺ, elevation: 1934 m). Four specimens were collected (SUHC 310313) (Fig. 2) and deposited in the Sabzevar University Herpetological Collection (SUHC), Iran. The sites of the new records near the border of Iran-Turkey are provided in Figure 3. Turkish specimens were collected from three different localities in 2010 and 2012 (Aydınlar Village, Adilcevaz, Bitlis—between Tatvan and Bitlis km 4, Bitlis—Hoşap, Güzelsu, Van). They were deposited in the Biology Laboratories of Dokuz Eylül University, İzmir, Turkey. DNA was obtained from tissue samples (muscle or liver) that were preserved in 96% ethanol (RastegarPouyani et al. 2010). DNA was extracted using a high salt method (Kabir et al. 2006). Fragments of Cytochrome b were ampliied (PCR) using the primers Mtanew (5ʹ-CTC CCA GCC CCA TCC AAC ATC TCA GGA TGA TGA AAC-3ʹ) and Mtfsh (5ʹ-TAG TTG GCC AAT GAT GAT GAA TGG GTG TTC TAC TGG-3ʹ). Sequences from Results Genetic distance (Table 2) indicates that the Iranian samples are very close to Eremias suphani of Turkey with only about 2% of genetic distance. Distances between Eremias strauchi population and both Eremias suphani and Eremias sp. samples from Iran, are 16%. Eremias sp. samples from Iran now conirmed as Eremias suphani. In addition to previous studies on the Iranian herpetofauna, our initial research in the NW corner of Iran revealed the presence of Eremias suphani in Iran. We focused our search in NW Iran to the border of Turkey for other localities, but in the other locations Eremias strauchi has been recorded. Table 1. Examined samples in this study along with their locality and coordinates. Code Species name N E Elevation Str-1 Eremias strauchi 39° 51′ 57.1′′ 42° 29′ 03.3′′ 864 m Locality Between Aralık and Gündoğdu, Iğdır Str-2 Eremias strauchi 39° 51′ 57.1′′ 42° 29′ 03.3′′ 864 m Between Aralık and Gündoğdu, Iğdır Str-3 Eremias strauchi 39° 51′ 57.1′′ 42° 29′ 03.3′′ 864 m SÜ2-A Eremias suphani 38° 18′ 18.2′′ 43° 48′ 46.4′′ 1,997 m Between Aralık and Gündoğdu, Iğdır Hoşap, Güzelsu, Van SÜ2-B Eremias suphani 38° 18′ 18.2′′ 43° 48′ 46.4′′ 1,997 m Hoşap, Güzelsu, Van SÜ2-C Eremias suphani 38° 18′ 18.2′′ 43° 48′ 46.4′′ 1,997 m Hoşap, Güzelsu, Van SÜ1-B Eremias suphani 38° 53′ 55.2′′ 42° 55′ 38.8′′ 2,000 m Aydınlar village, Adilcevaz, Bitlis SÜ1-C Eremias suphani 38° 53′ 55.2′′ 42° 55′ 38.8′′ 2,000 m Aydınlar village, Adilcevaz, Bitlis SUHC 724 Eremias sp. 38° 49′ 22.1′′ 44° 34′ 78.4′′ 1,934 m on the road from Firoragh to Chaldoran SUHC 725 Eremias sp. 38° 49′ 22.1′′ 44° 34′ 78.4′′ 1,934 m on the road from Firoragh to Chaldoran SUHC 726 Eremias sp. 38° 49′ 22.1′′ 44° 34′ 78.4′′ 1,934 m on the road from Firoragh to Chaldoran Amphib. Reptile Conserv. | amphibian-reptile-conservation.org 36 December 2013 | Volume 6 | Number 2 | e73 New country record and range extension of Eremias suphani Figure 2. General view of Eremias suphani from Iran. The color pattern of this species is different from Eremias strauchi strauchi (Photo by EskandarRastegar-Pouyani, 2005). genus Eremias were collected that were not compatible with diagnostic key of the Iranian species of Eremias, but were comparable to E. strauchi. Considering these observations we assumed that these specimens represented a new species of Eremias, and decided to compare the specimens with individuals of other species of the genus that were described and previously recorded from the region, speciically Eremias strauchi and Eremias suphani. Genetic analysis revealed that the Iranian samples clustered (with 2% distance) with E. suphani and not with E. strauchi (16% distance). This new country record demonstrates how the border regions of Iran have not been carefully investigated, and that additional effort must be made to document reptile diversity in these areas. These results strongly suggest that the Iranian samples are conspeciic with Eremias suphani. Consequently, we conclude that Eremias suphani is recorded for the irst time in Iran near the border with Turkey on the road from Firoragh to Chaldoran in Ali Sheykh village. Discussion Specimens of Eremias collected from Ahlat, Bitlis, in eastern Turkey were considered to be Eremias velox ssp. by Başoğlu and Hellmich (1959). In 1968, these authors examined specimens from Ahlat, Süphan Dağı, Mukus, and Van and described a new subspecies, Eremias velox suphani. Peters (1964) considered E. velox strauchi to be a full species (E. strauchi), whereas specimens collected from Doğubayazıt, Iğdır, and Kağızman were considered as E. velox by Clark and Clark (1973). Başoğlu and Baran (1977) stated that E. suphani and E. strauchi are subspecies of E. velox whereas Bischoff (1978) suggested that E. suphani is a subspecies of E. strauchi. Finally, E. velox suphani was regarded as a distinct species by Bishoff and Böhme (1980). Eremias suphani is considered to be endemic to eastern Turkey, where it is known from the vicinity of Lake Van (Mulder 1995; Franzen and Heckes 1999; Baran et al. 2012). According to the previous studies on the Iranian herpetofauna (Leviton et al. 1992; Anderson 1999; Rastegar-Pouyani et al. 2007, 2008), E. suphani has not been recorded from Iran. During ield trips to northwest Iran in 2005, four specimens from a population belonging to the Acknowledgments.—We thank Hamzeh Oraei and Azar Khosravani for their efforts in providing the molecular data and Dr. Jim Bogart for his generous help in improving the English of an earlier version of this manuscript. Table 2. Estimates of evolutionary divergence (p–distance) over sequence pairs between samples of Eremias in this study. Eremias suphani Turkey Eremias sp. Iran Firoragh Eremias suphani Turkey — 0.022 0.193 Eremias sp. Iran Firoragh 0.022 — 0.166 Eremias strauchi strauchi 0.193 0.166 — Amphib. Reptile Conserv. | amphibian-reptile-conservation.org 37 Eremias strauchi strauchi December 2013 | Volume 6 | Number 2 | e73 Rastegar-Pouyani et al. Figure 3. Iran-Turkey map and localities of new records in Iran and the type locality of Eremias suphani around the Van Lake in Turkey. Red circles relate to new country records in Iran and the blue circles are type localities of the Turkish samples. Fakültesi Kitaplar Series No. 76: 1–272. Bornova. Baran İ, Ilgaz Ç, Avcı A, Kumlutaş Y, Olgun K. 2012. Türkiye Amibi ve Sürüngenleri. TÜBİTAK, Ankara. 204 p. (ISBN: 978-975-403-703-6). Bischoff W. 1978. Beiträge zur Kenntnis der Echsen des Kaukasus. Salamandra 14: 178–202. Bischoff W, Böhme W. 1980. 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Amphibians and reptiles from Iran in the United State National Museum Collection. Bulletin of the Maryland Herpetological Society 7: 48–36. Tuck RG. 1974. Some amphibians and reptiles from Iran. Bulletin of the Maryland Herpetological Society 10: 59–65. Received: 03 November 2013 Accepted: 08 November 2013 Published: 14 December 2013 Appendix I. List of the material examined in this study and their accession numbers. Species name Country Accession Number Eremias suphani Turkey KF797802 Eremias suphani Turkey KF797803 Eremias suphani Turkey KF797804 Eremias suphani Turkey KF797805 Eremias suphani Iran KF797806 Eremias suphani Iran KF797807 Eremias suphani Iran KF797808 Eremias suphani Iran KF797809 Eremias strauchi strauchi Iran KF797810 Eremias strauchi strauchi Iran KF797811 Eremias strauchi strauchi Iran KF797812 Eremias strauchi strauchi Iran KF797813 Amphib. Reptile Conserv. | amphibian-reptile-conservation.org 39 December 2013 | Volume 6 | Number 2 | e73