Abstract
During a scientific field expedition to the Alai-Pamir range five specimens of the genus Gloydius have been collected in the larger Alai. A morphological and genetical examination of the specimens has shown that they are part of the G. halys complex, but represent a new taxon which is characterized by the following unique character combination: It is a slender and moderately stout small snake, up to 479 mm total length. The head has nine symmetrical plates on the upper head, 7 supralabial and 8-9 infralabial scales. Body scales in 20-22 rows around midbody, 143-156 ventral and 35-45 usually paired subcaudal scales. The cloacal plate not divided. The general coloration consists of various different tones of olive, tan and brown, having a distinct head, but an indistinct body pattern with, excluding the tail, 26-29 transverse crossbands, which are not extending to the sides of the body. The haplotype network shows the new species within the G. halys complex and close related to both, G. h. halys and G. h. caraganus. So far the new described species is only known from the Alai range. However, various Gloydius specimens are found in Kyrgyzstan and because of the complicated taxonomy those specimens have to re-identified to clarify their status and the status of the new species.
Introduction
Snakes of the genus Gloydius Hoge and Romano-Hoge, 1981 are venomous pitvipers and part of the Asian radiation of the subfamily Crotalinae within the Viperidae. Recently, 13 species are known: Gloydius blomhoffii (Boie, 1826) from China, Korea, Japan and Russian Far East (see Orlov et al., 2014); Gloydius brevicaudus (Stejneger, 1907) from China, North and South Korea; Gloydius halys (Pallas, 1776) ranging from Azerbaijan and Iran through several countries of Middle Asia to eastern Siberia, Mongolia, and China; Gloydius himalayanus (Günther, 1864) from northeastern Pakistan along the southern slopes of the Himalaya to northern India and Nepal; Gloydius intermedius (Strauch, 1868) restricted to Russian Far East, north-eastern China and the Korean Peninsula according to Orlov and Barabanov (1999). However, according to Gloyd and Conant (1990) as well as to Orlov in Ananjeva et al. (1998) G. intermedius is distributed from south-eastern Azerbaijan, through northern Iran, southern Turkmenistan, and north-western Afghanistan, to north-western China and Mongolia, while G. saxatilis (recognized as synonym of G. intermedius by Orlov and Barabanov [1999]) from eastern Siberia through northeastern China to North and South Korea); Gloydius lijianlii Jiang and Zhao, 2009 endemic to Daheishan Island, China; Gloydius liupanensis Liu, Song and Lua, 1989 only known from its type locality in the Liupan Mountains of China; Gloydius monticola (Werner, 1922) only known from mountains in northern Yunnan, China; Gloydius shedaoensis (Zhao, 1979) endemic to Shedao Island, China; Gloydius strauchi (Bedriaga, 1912) known from the Tibetan Plateau of “Tsinghai” (= Quinghai) and western “Szechwan” (= Sichuan), China; Gloydius tsushimaensis (Isogawa, Moriya and Mitsui, 1994) endemic to Tsushima Island, Japan; and Gloydius ussuriensis (Emelianov, 1929) ranging from far east Russia through north-eastern China to North and South Korea, as well as on Quelpart Island.
Gloydius was the subject of considerable taxonomic instability. Gloyd and Conant (1990) published an intensive review of the genus Agkistrodon Palisot de Beauvois, 1799 (including Gloydius at this time) providing intensive morphological data and a new systematic approach for this group. A few years later, Bour (1993), based on his interpretation of the type locality of the nominate subspecies, synonymized G. h. caraganus with G. h. halys and introduced the name Agkistrodon (= Gloydius) halys mogoi for the populations of the former nominate taxon. However, Chernov (1934), Ananjeva et al. (1997) as well as Orlov and Barabanov (1999) did not accept this restriction. They revalidated G. h. caraganus, synonymized G. h. mogoi with G. h. halys and stabilized the latter with the designation of a neotype. Moreover, Orlov and Barabanov (1999) radically revised the classification made by Gloyd and Conant (1990) and did not accept G. halys and G. intermedius as two widely sympatric species across Central Asia as suggested by Gloyd and Conant (1990). Instead, they restricted G. intermedius to eastern Asia and suggested a single, widespread G. halys including several subspecies: (1) G. h. boehmei (Nilson, 1983); (2) G. h. caraganus (Eichwald, 1831); (3) G. h. caucasicus (Nikolsky, 1916); (4) G. h. cognatus (Gloyd, 1977); (5) G. h. halys (Pallas, 1776); and (6) G. h. stejnegeri (Rendahl, 1933). A taxonomic concept followed by Gumprecht et al. (2004). Additionally, the re-examination of the lectotype of G. intermedius by Orlov and Barabanov (1999) has shown that it is identical with Gloydius saxatilis. Consequently, they synonymized G. saxatilis with G. intermedius. Later, Xu et al. (2012), following the taxonomic concept of Gloyd and Conant (1990), published a phylogenetic study, focused on the Chinese species of the genus and showing G. intermedius basal to a lineage including G. saxatilis/G. shedaoensis and their sister species G. halys. Most recently, Hoser (2013) nomenclaturally revised the entire Crotalinae and introduced the generic name “Conantvipera” for some Gloydius species. However, this author is heavily criticized by the entire herpetological community (see e.g., Kaiser et al., 2013) for his nomenclatural acts which are published in his self-made journal, without any reliable research background and do not fulfill the criteria of the code of ethics. Therefore, the name introduced by Hoser (2013) is ignored in this publication (although LB consider them as formally available and valid according to the Code of Zoological Nomenclature until the International Commission on Zoological Nomenclature consider them as invalid).
This summary shows that Gloydius is a taxonomic complex group and, despite the intensive reviews by Gloyd and Conant (1990) and Orlov and Barabanov (1999) and the phylogenetic study by Xu et al. (2012), an integrative revision is in need to clarify the relationships between the species, their distributions and especially to clarify the status of the huge number of recognized subspecies. Herein, we follow the taxonomic concept of Orlov and Barabanov (1999), with some modification concerning the composition of the Gloydius halys complex and the usage of the genus name Gloydius.
Although many expeditions, started from Fedchenko, Severtsov and others, provided various herpetological collections and data, no pitvipers were recorded in the Alai region until Karpenko (1957). Later, these snakes were recorded in several localities of the Alai region (Yakovleva, 1964; Eremchenko, 2007). In 2013, an international research expedition was conducted to the Alai and Pamir region and was based for several days in a small valley leading into the larger of the Alai valley. Despite the fact, that this is a famous area and well-known for mammal and bird wildlife, the knowledge about the other vertebrate groups is limited. It is based on the results of William Frederik Reinig, the zoologist of the German-Soviet Alai-Pamir expedition which was organized by Willi Rickmer Rickmers in 1928 (see e.g., Reinig, 1928, 1932; Rickmer Rickmers, 1929, 1930). However, Reinig was mainly focused on arthropods and especially the herpetofauna of the Alai and Pamir is still only rudimentarily known. Therefore, collections of amphibians and reptiles from this area are important and done by the expedition.
The Pamir and Alai range is ecologically characterized by high altitude steppes with very pronounced temperature variation between summer and winter and low precipitation throughout the year. The Pamir has great altitudinal variation ranging from 1740 to 7495 m a.s.l., while the Alai range only up to 5544 m. However, both are characterized by high atmospheric aridity, intense insolation, great seasonal and large diurnal temperature fluctuations, low temperatures, scanty of no snow cover and cold storms (Agakhanyantz and Lopatin, 1978; Mani, 2007). The Pamir Mountains and the Alai Valley are among the priority areas for nature conservation. They are known as hotspot for mountain species of Central Asia and the diversity includes endemic plant, insect, amphibian, reptile and mammal species. Furthermore, a study of the Conservation International Foundation (2014) at the Pamir-Alai Transboundary Conservation Area estimated that up to 51% of the invertebrate fauna is endemic to the region, while Karamhudoeva (2008) recognized about 15% of the Lepidoptera species as endemic for the Pamir and Alai range. In Gloydius halys, the Alai is of special interest because according to Orlov and Barabanov (1999) it is an area where the ranges of the two subspecies G. h. halys and G. h. caraganus are overlapping. However, according to Gloyd and Conant (1990) only the latter is present in the Alai, but in its extreme southeastern most distribution.
Therefore, the specimens were examined in details and compared with other subspecies of G. halys and the results are presented in this publication.
Material and methods
The specimens were compared with the morphological data provided by Gloyd and Conant (1990) and Orlov and Barabanov (1999) and furthermore with Gloydius specimens housed in the ZFMK collection (see Appendix). The following institute acronyms are used in this publication: ANSP, Academy of Sciences in Philadelphia, PA, USA; BMNH, The Natural History Museum, London, England; ISEA, Institute of Systematics and Ecology of Animals, Siberian Branch of Russian Academy of Sciences, Novosibirsk, Russia; MNHN, Muséum National d’Histoire Naturelle, Paris, France; MNKNU, Museum of Nature at V. N. Karazin Kharkiv National University, Kharkiv, Ukraine; NHRM, Naturhistoriska Riksmuseet, Stockholm, Sweden; NMW, Naturhistorisches Museum, Wien, Austria; USNM, United States National Museum, Washington, DC, USA; UTA, University of Texas in Arlington, Merriam Museum, Arlington, TX, USA; ZMB, Museum für Naturkunde, Berlin, Germany; ZFMK, Zoologisches Forschungsmuseum Alexander Koenig, Bonn, Germany; ZISP, Zoological Institute, Russian Academy of Sciences, St. Petersburg, Russia.
Tissue samples used in this study with locality, voucher and GenBank accession numbers.
Haplotype network of selected members of Gloydius halys/intermedius complex based on ND4 sequences. Numbers below “l” corresponds to the number of mutated positions (“l” without number corresponds to single mutation).
Citation: Amphibia-Reptilia 37, 1 (2016) ; 10.1163/15685381-00003026
For the comparison the following characters were used and compared with the above mentioned sources: head length, head width, head height, snout-vent length, tail length (all taken with a digital caliper to the nearest of 0.1 mm); number of supralabial, infralabial, temporal, ventral and subcaudal scales; shape of head, body and tail scales; number of scale rows at fore body (one head length posterior to the head), midbody and hind body (equivalent of one head length anterior to the cloacal plate), all counted straight across the body; number of crossbands on the body excluding the tail, width of crossbands counted as scale rows, extension of crossbands referring to the midbody scale row counted from the ventral scales.
Digital X-ray images for skull illustration of the specimen ZMB 80360 were done using a Faxitron LX-60 Closed Cabinet Digital Specimen Radiography System (Faxitron Corp.) at the ZFMK. Figures used for X-ray comparisons are modified from Gloyd and Conant (1990): G. blomhoffii blomhoffii (KVK 832 without locality), G. intermedius (ANSP 30146 from Adsh Mountain Range, Mongolia [according to Orlov and Barabanov (1999) this specimen correspondents with G. h. halys]), G. halys caraganus (ZIK 292 from near Kulsari, Kazakhstan), and G. himalayanus (BMNH 1930.5.8.971 from Bakloh, India).
Relationships of the Alai specimens and other taxa of the G. halys-intermedius complex were inferred using partial nucleotide sequences of the mitochondrial gene NADH dehydrogenase subunit 4 (ND4). Total genomic DNA was isolated from the liver or muscle tissues of the ethanol stored museum specimens using standard proteinase K and phenol-chloroform protocols (Sambrook, et al., 1989; see table 1 for specimens and Genbank numbers). A target fragment was amplified using primers Nd4 and tRNA-leu (Arevalo, et al., 1994). PCR was performed in a 25 μl volume each containing 15-60 ng DNA, 0.2 mM of each dNTP, 2.5 μl of 10× amplification buffer (10 mM Tris-HCl pH 8.5, 50 mM KCl and 2.5 mM MgCl2), 1 U of Taq DNA polymerase, and 5 pmol of primers. Amplification was performed in a T100™ Thermal Cycler (Bio-Rad) with an initial denaturation step for 3 min at 94°C, followed by 35 cycles of 60 s at 94°C, annealing at 54°C for 30 s, 60 s extension at 72°C, and a final extension of 5 min at 72°C. Sequencing was carried out on an ABI 3730 automated capillary sequencer (Applied Biosystems) with the ABI Prism Big Dye Terminator Cycle Sequencing Ready Reaction Kit 3.1 using the same primers.
DNA sequences were aligned manually and checked for unexpected stop codons using BioEdit 7.0 (Hall, 1999). Aligned sequences were 641 bp long. MEGA 6 (Tamura, et al., 2013) was used to calculate percentage differences (p-distances) between obtained sequences. A haplotype network was generated using the median-joining algorithm (Bandelt et al., 1999) implemented in Network 4.6.1.2 (www.fluxus-engineering.com).
Results
The morphological comparison revealed differences in pholidosis to other Gloydius taxa. Following the key presented by Gloyd and Conant (1990) the Alai specimens would refer to G. monticola or G. h. cognatus. However, both taxa range far away from the Alai valley and are superficially similar but very distinct in detail. The specimens are distinct to most Gloydius taxa by their low number of scale rows around midbody, which is only similar in the combination of the missing apical pits to G. monticola. But, the Alai specimens have a higher number of scale rows around midbody and a very distinct coloration. Recognizing the low number as unusual but within the variation, in combination of the missing apical pits, this would refer, because of the low numbers of ventral scales and body crossbands to G. h. cognatus. However, from this taxon the Alai specimens are distinct in coloration having body crossbands not extending to the lateral sides of the body and in some aspects of scalation (see below).
Nucleotide sequences of the ND4 gene were obtained for two Alai specimens, G. h. halys, G. h. caraganus and G. intermedius (table 1). Both Alai specimens shared the same haplotype and were fairly distinct from G. halys and other taxa sequenced in this study (fig. 1). Consequently the specimens are described as a new species.
Account of related taxa
The species of the genus Gloydius are recognized here in the sense of Orlov and Barabanov (1999, see introduction for comments) and all of the following species are recognized herein as members of the G. halys complex, a subgroup mainly including Central and Eastern Asian taxa.
Gloydius halys halys (Pallas, 1776)
- 1776 Coluber halys Pallas, Reise durch verschiedene Provinzen des russischen Reichs. Kais. Akad. Wiss., St. Petersburg, Vol. 3: 703.
- 1820 Echidna aspis var. pallasii Merrem, Versuch eines Systems der Amphibien I (Tentamen Systematis Amphibiorum). J. C. Kriegeri, Marburg: 151.
- 1993 Gloydius halys mogoi Bour, Les voyages de Peter Simon Pallas et l’origine de Coluber halys (Serpentes Viperidae). Bull. mens. Soc. linn. Lyon 62: 395.
Neotype. ZISP 14784, an adult male from the Borgaiskaya steep, 84 km West from Kyakhta town, Burin-Khan mountain (designated by Orlov and Barabanov, 2000).
Diagnosis. A moderately stout viper up to 530 mm total length in males, to 590 mm in females according to Gloyd and Conant (1990), but up to 750 mm fide Orlov and Barabanov (1999). Snout seen in profile recurved; supralabial scales usually 7-9. Apical pits absent. Dorsal scales in 23 (very rarely in 21 or 25) rows around midbody; ventral scales between 164-178 (147-187 fide Orlov and Barabanov, 1999); subcaudal scales paired, between 42-49 (29-56 fide Orlov and Barabanov, 1999). Body with 33-47 dark transverse bands, each 3-5 scales wide and extending down to scale row 3 or 2; light areas between blotches relatively narrow. Light line above dark cheek stripe on 1-1.5 adjacent rows of scales.
Distribution. Gloyd and Conant (1990) and Orlov and Barabanov (1999) mentioned a distribution from eastern Kazakhstan through southern Siberia to Mongolia and a vertical distribution between 150 to at least 2300 m.
Gloydius halys boehmei (Nilson, 1983)
- 1983 Agkistrodon halys boehmei Nilson, A new subspecies of the Asiatic pit viper Agkistrodon halys Pallas, 1776 (Serpentes, Viperidae). Bonner zoologische Beiträge 34: 470.
Holotype. ZFMK 8648, from the Andarab Valley in the province Baghlan, Afghanistan.
Diagnosis. A small viper up to 487 mm total length. Apical pits absent. Two pre- and two postocular scales on each side. Seven supralabial scales and 11 sublabial scales. Dorsal scales in 23 rows around midbody; ventral scales 155; 35 subcaudal scales paired, cloacal plate not divided. Body with 41 dark transverse bands, each 3-4 scales wide and extending down to scale row 7 or 8.
Distribution. Only known from its type locality at 2500 m a.s.l.
Gloydius halys caraganus (Eichwald, 1831)
- 1831 Trigonocephalus caraganus Eichwald, Zoologia specialis, quam expositis animalibus tum vivis, tum fossilibus potissimuni rossiae in universum, et poloniae in specie, in usum lectionum publicarum in Universitate Caesarea Vilnensi. Zawadski, Vilnae: 170.
- 1931 Ancistrodon halys paramonovi Nikolsky, Eine neue Schlangenunterart aus Turkestan [Ukrainian with German title], Travaux Mus. Zool. (Kiev) 10: 115. Type locality: “in montibus Tschimganensis altitudine 1500-2500 m probe urbem Taschkent in Turkestano” (= mountains near Tashkent, Uzbekistan).
Neotype. ZISP 2200, adult male from the Mangyshlak Peninsula, eastern edge of the Caspian Sea, Kazakhstan (designated by Orlov and Barabanov, 1999).
Diagnosis. A relatively slender and moderately stout viper up to 735 mm total length in males (740 mm and more according to Orlov and Barabanov, 1999), to 530 mm in females. Snout seen in profile slightly recurved, supralabial scales usually 8 (71%) sometimes 7. Apical pits absent. Dorsal scales in 23 (rarely 21) rows around midbody; ventral scales between 149-167 (141-183 fide Orlov and Barabanov, 1999); subcaudal scales paired, between 33-47 (16-51 fide Orlov and Barabanov, 1999). General coloration is pale. Body with 36-50 dark transverse bands, not extending low on the sides and with relatively broad light areas between them.
Distribution. From northeastern shores of the Caspian Sea through Kazakhstan, Uzbekistan, northwestern Tadzhikistan and Kirgizia to Chinese border and probably extreme western China.
Remarks. Orlov and Barabanov (1999) as well as Gumprecht et al. (2004) mentioned G. h. halys and G. h. caraganus as sympatric in parts of their distribution range. Therefore this subspecies should be recognized on species rank, probably including other G. halys subspecies which needs to be clarified by further research.
Gloydius halys caucasicus (Nikolsky, 1916)
- 1916 Ancistrodon halys caucasicus Nikolsky, Ophidia. Fauna of Russia and adjacent countries. Vol 2. Reptiles: 267 [in Russian].
- 1933 Ancistrodon halys persicus Rendahl, Die Unterarten des Ancistrodon halys Pall. Nebst einigen Bemerkungen zur Herpetologie Zentralasiens. Arch. Zool. (Stockholm) 25A (8): 11.
Neotype. ZISP 19017.1, adult male from the vicinity of Kirovsk town, Lenkoran district, Azerbaijan (designated by Orlov and Barabanov, 1999).
Diagnosis. A moderately stout viper up to 660 mm total length. Supralabial scales 7-8, rarely 9. Dorsal scales in 23 (rarely 25) rows around midbody; ventral scales between 142-169; subcaudal scales paired, between 31-46. Body with 33-42 dark transverse bands, each 4-6 scales wide and extending down to scale row 3.
Distribution. The subspecies is ranging from southeastern Azerbaijan through northern Iran to southern Turkmenistan (Kopet Dag Mountains) and northwestern Afghanistan (see Orlov and Barabanov, 1999; Wagner et al., in rev.).
Gloydius halys cognatus (Gloyd, 1977)
- 1977 Agkistrodon halys cognatus Gloyd, Descriptions of new taxa of crotalid snakes from China and Ceylon (Sri Lanka). Proc. Biol. Soc. Washington 90: 1002.
Holotype. USNM 68586, from Choni on the Tao River, Gansu province, China.
Diagnosis. A small viper of the genus, up to 590 mm total length. Supralabial scales 7-8. Dorsal scales in 23 (rarely 21) rows around midbody; ventral scales between 153-165; subcaudal scales paired, between 36-54. Body with 29-43 dark transverse bands, each 4-5 scales wide and extending down to scale row 4 to 2.
Distribution. Recently endemic to northern Central China, but probably northwards into Inner Mongolia.
Gloydius halys stejnegeri (Rendahl, 1933)
- 1933 Ancistrodon halys stejnegeri Rendahl, Die Unterarten des Ancistrodon halys Pall. Nebst einigen Bemerkungen zur Herpetologie Zentralasiens. Arch. Zool. (Stockholm) 25A (8): 18.
Lectotype. NHRM 1923 809.2780, from China.
Diagnosis. A small viper of the genus, up to 625 mm total length. Supralabial scales 7-8. Dorsal scales in 23 rows around midbody; ventral scales between 147-165; subcaudal scales paired, between 39-46. Body with 28-38 dark transverse bands, each 3-6 scales wide and extending down to scale row 3 or 2.
Distribution. The taxon ranges from the edge of the Gobi in southeastern Inner Mongolia southwards to the Shanxi and Hebei provinces in China (Gloyd and Conant, 1990).
Gloydius intermedius (Strauch, 1868)
- 1868 Trigonocephalus intermedius Strauch, Concerning poisonous snakes distributed in Russia. Trudy Perv. Siezda Russ. Yestestv. Zool. 1: 294 [in Russian].
Lectotype. ZISP 2221, from “Prom. Tyr (Amur) [= Cape Tyr, Amur River, Amurskaya Region, Russian Far East]” (designated by Orlov and Barabanov, 2000).
Diagnosis. A large viper of the genus, up to at least 800 mm total length. Supralabial scales 7-8. Dorsal scales in 23, very rarely 21, rows around midbody; ventral scales between 148-175; subcaudal scales paired, between 34-52. Body with 28-45 dark transverse bands, each 3-6 scales wide and extending down to scale row 2 or 1.
Distribution. According to Orlov and Barabanov (1999) the species is restricted to an area from Russian Far East through northeastern China to the Korean Peninsula.
Gloydius rickmersi sp. n. Wagner, Tiutenko, Borkin and Simonov
Holotype. ZMB 80360 [field no. PW P069], adult female from Kul-Otek at the Sary-Buka Valley in Kyrgyzstan, about 25 km (by air) NE of Daroot-Korgon (Chong-Alai District) near the town Kyzyl-Eshme in direction to the Shuman-Bol pass and the Kichi-Alai River at an elevation of 3000 m a.s.l. [N39.622132, E72.284010], collected on 4.VIII.2013, at about 19.00 h and purchased by local boys to Philipp Wagner. The new name was registered at Zoobank under the LSID: 84644B3D-85BC-4BF6-8EDF-925D8E444470.
Paratypes. MHNG 2745.46 (field no. PW P070), two pieces of a subadult female from the same area as the holotype but about 200 m higher in elevation, collected as dead body on 7.VIII.2013 by Philipp Wagner. MHNG 2752.70, roadkilled at Kul-Otek near to one from Kyzyl-Eshme, collected as dead body by Glib Mazepa. MHNG 2752.69, roadkilled at the northern slope of the Alai ridge, Tengizbai River, Kichelay [N39.755, E72.202], 2798 m, collected by Spartak Litvinchuk. ZSM 211/2014, a juvenile male from the same area as the holotype but lower in elevation towards the entrance of the valley to the main Alai valley, collected as dead body on the road on 16.VIII.2013 by Arthur Tiutenko.
Diagnosis. A slender, moderately stout small snake of the genus Gloydius, with up to 479 mm total length. Head slender, slightly triangular, with nine symmetrical plates on the upper head, 7 supralabial and 8-9 infralabial scales. Body scales in 20-22 rows around midbody, 143-156 ventral and 35-45 usually paired subcaudal scales. Cloacal plate not divided. Body, excluding the tail, with 26-29 transverse crossbands, not extending to the sides of the body.
Description. A small snake of its genus, but only one adult specimen is recently known. Head slender, slightly triangular and with nine symmetrical plates on top. Snout slightly upturned. Parietal scales large, longer than wide. Two nasal scales present, with the nostril pierced in the center between them. Loreal scale about quadrangular. Six scales surrounding the orbit, excluding the postfoveal. Two preocular, two postocular and three temporal scales. Pit bordered anteriorly by a large prefoveal, dorsally and ventrally by a narrow prefoveal, and posteriorly by the angle of both of them. Seven sublabial scales on each side, the second is the smallest, the third is entering the orbit. Eight to nine infralabial scales. Body scales elongated and mainly keeled, the first two scale rows bordering the abdominal plates mooth at midbody. Apical pits absent. Body scales arranged in 22 [anterior]-20-22 [midbody]-15-17 [posterior] rows around the body. There are 143-156 ventral scales, excluding the terminal scale. Cloacal plate not divided. 35-45 subcaudal scales usually paired, but sometimes the first two thirds of the tail bear unpaired, followed by paired scales.
Differential diagnosis. The new species is distinct to all other species of the genus in the combination of the following characters: a slightly lower number of scale rows around midbody (20-22 versus mainly 23, sometimes 21); a low number of transverse body crossbands (26-29 versus above 30 in all taxa beside G. himalayanus and the G. blomhoffii complex), the mean number of ventral scales (150 which is higher than in most of the G. blomhoffi taxa, but distinctly lower than in the G. halys-intermedius complex); and its small size (479 mm total length versus more than 600 mm is most species, apart from G. b. dubitatus, G. b. ussuriensis, G. h. halys and G. h. caucasicus which are about the same maximum total length, and G. monticola and G. strauchi which are smaller). In details, the new species is distinct to taxa of the Gloydius blomhoffi complex generally by the lack of apical scale pits, but as well by having a higher number of ventral scales (mean 150 versus 140.7 in G. b. blomhoffii, 140.1 in G. b. dubitatus, 141.1 in G. b. brevicaudus, and 140.1 in G. b. siniticus; but apart from G. ussuriensis with 151.2); in having a lower number of infralabial scales (mean 8.5 versus 10.4 in G. b. blomhoffii, 10.0 in G. b. dubitatus, 10.0 in G. b. brevicaudus, 10.1 in G. b. siniticus, and 9.8 in G. ussuriensis); in having a higher or somewhat equal number of crossbands on the body (mean 27.5 versus 19.9 in G. b. blomhoffii, 29.9 in G. b. dubitatus, 29.9 in G. b. brevicaudus, 30.1 in G. b. siniticus, and 28.7 in G. ussuriensis); and by having a very indistinct coloration of crossbands versus very distinct crossbands in this complex. The new species is distinct to taxa of the Gloydius intermedius complex (according to the concept of Gloyd and Conant, 1990) by the absence of apical scale pits; in having a lower number of scale rows around midbody (mean 21 versus 22.8 in G. intermedius and G. saxatilis); in having a lower number of ventral scales (mean 150 versus 158.5 in G. intermedius and 156.8 in G. saxatilis); and a lower number of body crossbands (mean 27.5 versus 36.9 in G. intermedius and 38.7 in G. saxatilis). Furthermore, the new species is distinct to all taxa of the Gloydius halys complex in having a lower number of scale rows around midbody (20-22 [mean = 21] versus usually 23 rarely 21 or 25), and very narrow indistinct crossbars along the body not extending to the lateral sides (versus distinct crossbars and extending to the flanks in most taxa) (see table 2). The new species is distinct to G. h. halys by a lower number of ventral scales (143-156, mean 150 versus 164-178, mean 172); by a lower number of infralabial scales (mean 8.5 versus 10.7); a lower number of body crossbands (mean 27.5 versus 38.4); and a distinct coloration (body crossbands not extending to the flanks versus extending to the flanks). The new species is distinct to G. h. boehmei by its higher number of subcaudal scales (35-45 versus 35); its lower number of infralabial scales (mean 8.5 versus 11); and a lower number of body crossbands (mean 27.5 versus 41). Apart from the above mentioned characters, the new species is somewhat similar in coloration to G. h. caraganus, but distinct in its lower number of ventral scales (143-156, mean 150 versus 149-167, mean 158); by its lower number of infralabial scales (mean 8.5 versus 10.8); and by its lower number of body crossbands (mean 27.5 versus 44.4). The new species is distinct to G. h. caucasicus by its slightly lower number of ventral scales (mean 150 versus 157); the lower number of infralabial scales (mean 8.5 versus 11); a lower number of body crossbands (mean 27.5 versus 36.4); a distinct coloration (body crossbands not extending to the flanks versus extending to the flanks); and by the absence of apical scale pits. The new species is distinct to G. h. cognatus by its slightly lower number of ventral scales (mean 150 versus 159); the lower number of infralabial scales (mean 8.5 versus 10.2); a lower number of body crossbands (mean 27.5 versus 35.9); and a distinct coloration (body crossbands not extending to the flanks versus extending to the flanks). The new species is distinct to G. h. stejnegeri by slightly lower number of ventral scales (mean 150 versus 156); the lower number of infralabial scales (mean 8.5 versus 10.7); a lower number of body crossbands (mean 27.5 versus 31.6); a distinct coloration (body crossbands not extending to the flanks versus extending to the flanks); and by the absence of apical scale pits.
Variation in selected Gloydius species. Values in the new species refer to this study and include the range followed by the mean value in brackets. Values of other taxa refer above to Gloyd and Conant (1990), followed by their mean value in brackets, while values below refer to Orlov and Barabanov (1999). Own data in square brackets. AP: apical pits; CP: cloacal plate; IL: infralabial scales; o: or; r: rarely; SC: subcaudal scales; SM: scales around midbody; SL: sublabial scales; ToL: total length (in mm); TVB: transversal bands; V: ventral scales; ∗the holotype has paired/unpaired subcaudal scales.
Holotype (ZMB 80360) of Gloydius rickmersi sp. n. from Sary-Buka Valley, Kyrgyz Republic. This figure is published in colour in the online version.
Citation: Amphibia-Reptilia 37, 1 (2016) ; 10.1163/15685381-00003026
Detailed pholidosis of the holotype (ZMB 80360) of Gloydius rickmersi sp. n. from Sary-Buka Valley, Kyrgyz Republic.
Citation: Amphibia-Reptilia 37, 1 (2016) ; 10.1163/15685381-00003026
Description of the holotype. A small and slender adult male (figs 2, 3) with a total length of 479 mm (body length 414.5 mm, tail length 64.5 mm). Head slender, slightly triangular, 18 mm in length, 13.35 mm wide and 8.2 mm heigh. Crown with nine symmetrical plates. Internasal scales more than two times wider than long, their posterior margins curving slightly backwards. Internasal and prefrontal scales somewhat thickened, and with a slight depression of the suture between the frontal and prefrontal as well the prefrontal and internasal scales, giving the appearance of a slightly upturned snout. Parietal scales large, about 1.5 times longer than wide. Rostral slightly higher than wide, with an obtuse angle above. Two nasal scales present, with the nostril pierced in the center between them. Anterior scale larger than the posterior one. Loreal scale subquadrangular, slightly wider than high. Six scales surrounding the orbit, excluding the postfoveal. Two preocular scales, the lower one narrow and forming the posterior dorsal border of the pit. Two postocular scales, the lower one crescent-shapes and extending forward beneath the eye. Pit bordered anteriorly by a large prefoveal, dorsally and ventrally by a narrow prefoveal, and posteriorly by the angle of both of them. Seven sublabial scales on each side, the second is distinctly smaller than the others, the third is entering the orbit. Eight infralabial scales on the right, and nine on the left side. Temporal scales form a horizontal row and consist of three scales on each side, decreasing in size posteriorly. Medium-sized mental scale, triangular, sides about equal. First pair of infralabial scales in short contact behind the mental. One pair of chin shields, 1.5 times longer than wide. Gular scales along the median from chin shields to first ventral scale in five rows, from infralabial to infralabial in ten rows. Body scales elongated and mainly keeled. The keel is in the centre and nearly as long as the scale. Apical pits absent. The first two scale rows bordering the abdominal shields smooth, the second row becoming keeled at the last third of the body towards the tail. Body scales arranged in 22 rows interiorly (at ventral scale no. 15), in 21 rows at midbody (no. 79 = half of the SVL) and in 17 rows posteriorly (no. 143). There are 149 ventral scales, and additional two preventral scales. Cloacal plate not divided. First 18 subcaudal scales unpaired (totally two third of the tail length), followed by 24 paired scales, resulting in a total number of 42 subcaudal scales.
Coloration. The general coloration consists of various different tones of olive, tan and brown. The head pattern is distinct, while the body pattern is very vague. Pattern of the head with a dark brown, somewhat triangular spot situated medianly on the internasal and prefrontal scales. Followed by a pair of somewhat quadrangular spots, nearly in contact to each other, and one on each side of the head, occupying most of the supraocular scales posteriorly and parts of the frontal and parietal scales. A second pair of rhomboidal spots, not nearly in contact, is situated on the posterior part of the parietal scales. This pair is followed by a pair of elongated blotches on the back of the head, becoming confluent posteriorly and enclosing a sand-hour like light area in the median area of the hind head. A dark brown, somewhat white bordered, cheek stripe extends from the orbit to beyond the angle of the jaw. The upper white line occupies the lower edges of an adjacent row of horizontal temporal scales, while the lower extends from the lower postocular across the lower edge of the first temporal scale and the last three supralabial scales. Supralabial scales speckled with gray, infralabial scales dark brown with whitish spots. The body coloration consists of a series of very indistinct, irregular, often incomplete, and narrow pale olive dorsal crossbands which are incompletely and indistinctly dark brown dark bordered and do not extend to the lateral sides of the body. Crossbands longitudinal along the median line two, rarely three, scale rows long, and across the body seven scale rows wide. Totally in 29 crossbands along the body, and in 6 along the tail. The belly is dirty white with a gray speckling. A series of dark brown regular ventrolateral blotches is present on each side along the entire body.
Digital X-ray image (left: dorsal; right: lateral view) of the holotype of Gloydius rickmersi sp. n. (ZMB 80360), compared with figures modified from Gloyd and Conant (1990). For further details see Material and methods.
Citation: Amphibia-Reptilia 37, 1 (2016) ; 10.1163/15685381-00003026
Skull. The skull of Gloydius rickmersi sp. n. is overall similar to the skulls of G. h. halys and G. h. caraganus (fig. 4), and distinct to G. blomhoffii and G. himalayanus which have more elongated and flat skulls. It is similar to G. h. caraganus in the shape of the premaxilla bones and the elongated quadrate, but distinct in the dentition of the lower maxilla and the wider angle of the quadrate bones. In the latter characters it resembles G. h. halys from which it is distinct in the shape of the premaxilla. The dentition of the upper jaw is similar to taxa of the G. halys complex but distinct to G. blomhoffi which has much smaller teeth. The new species has more teeth in the lower jaw than G. h. caraganus but a similar number as compared with G. h. halys.
Variation. Both rather complete paratypes generally agree with the holotype. MHNG 2745.46 has a body length of 371 mm and a tail length of 56 mm. The head scalation is identical to the holotype, but the infralabial scales are not countable. There are two preventral, 154 ventral and 35 subcaudal scales which are entirely paired. Body scales around the body in 21 rows at the anterior, in 20 rows at the midbody and 15 rows at the posterior portion of the body. The 26 body and 4 tail crossbands are much more indistinct and narrow than in the holotype. ZSM 211/2014 has a body length of 312 mm and a tail length of 71 mm. The scalation is identical with the holotype but differs in the following aspects: There are 9 infralabial scales on both sides, two preventral, 141 ventral and 45 entirely paired subcaudal scales. Body scales around the body in 22 rows at the anterior, in 22 rows at the midbody and 17 rows at the posterior portion of the body. Body and tail coloration similar to the holotype, but the crossbands are too indistinct and not countable. The belly coloration is uniform cream, lacking the gray speckling and dark brown blotches as in the holotype.
Distribution of Gloydius rickmersi sp. n. in the Kyrgyz Republic. 1: Kul-Otek at the Sary-Buka Valley about 25 km (by air) NE of Daroot-Korgon (Chong-Alai District) near the town Kyzyl-Eshme in direction to the Shuman-Bol pass and the Kichi-Alai River. 2: Northern slope of the Alai range, Tengizbai River, Kichelay, 2798 m.
Citation: Amphibia-Reptilia 37, 1 (2016) ; 10.1163/15685381-00003026
Distribution. So far the new species is only known from its type locality and one locality at the northern slope of Alai range (Kichelay at the Tengizbay River, beyond the Tengizbay Pass, see fig. 5). However, within the small valley of the type locality it has a broader distribution and occurs from the entrance of the valley near the Alai lowlands, up to the end of the valley in higher elevation. A specimen recognized as G. halys from “Daraut River Canyon” by the PATCA Project most probably refers to the herein described new species. Other specimens recognized from eastern parts of the Alai and from the entire country have to be re-examined to clarify their identity. Orlov and Barabanov (1999) mentioned the altitudinal distribution of G. h. halys and G. h. caraganus as distinct. While the previous one is referred to elevations up to 3000 m, the latter is mentioned as inhabiting deserts and hilly habitats not higher than 1000 m a.s.l. However, Gloyd and Conant (1990) are providing different ranges with G. h. caraganus ranging up to at least 4000 m and G. h. halys between 150 and 2300 m. Gloydius rickmersi sp. n. was collected between 2800-3000 m a.s.l. and therefore within the range of G. h. caraganus but higher than of G. h. halys according to Gloyd and Conant (1990). However, the upper end of the range of G. h. halys is much higher than G. h. caraganus following Orlov and Barabanov (1999).
Relationships. The haplotype network (fig. 1) shows G. rickmersi sp. n. within the G. halys complex. It is close related to both, G. h. halys supported by the p-distances of 4.8-5.0%, and to G. h. caraganus showing a not considerably higher p-distance of 5.1%. In comparison, the p-distance between G. h. halys and G. intermedius is only 3.6%.
Habitat of Gloydius rickmersi sp. n. at the type locality in the Sary-Buka Valley. This figure is published in colour in the online version.
Citation: Amphibia-Reptilia 37, 1 (2016) ; 10.1163/15685381-00003026
Habitat and ecology. The holotype was collected at dusk on the road slightly above the base camp (fig. 6), while the topotypical paratype (MHNG 2745.46) was collected as dead body, probably killed by a predator next to a small stream about 200 m above the base camp. According to Yakovleva (1964) Gloydius h. caraganus is very variable in Kirgizia regarding the habitat preference and specimens were found on plains, in steppe foothills, meadows, open shrubby areas, the rocky banks of rivers, on mountain slopes and open forests. However, according to both Gloyd and Conant (1990) and Orlov and Barabanov (1999) not all Gloydius specimens in Kirgizia can be recognized as G. h. caraganus and therefore the mentioned habitats could refer to a variety of taxa. It can be assumed that the herein described species does not has a similar variability, and is restricted to higher altitudinal habitats but should inhabit both, the rocky and drier areas as well as the meadows in the valley. Two dead bodies were found directly next to small streams, which supports the wet meadows as habitat as well. Several old dead bodies were found on the road along the valley and it can be suggested that the road is frequently used for body-temperature regulation before the night. Especially as the holotype was collected at dusk and according to reports by local people, G. rickmersi sp. n. seems to be nocturnal.
At the type locality Asymblepharus alaicus yakovlevae Eremchenko, 1983 as well as Bufo pewzowi Bedriaga, 1898 were recognized. In the habitat of the specimen from the northern slope of the Alay Range Eremias nikolskii Nikolsky, 1905 and again Bufo pewzowi were found.
Etymology. Nearly 50 years after his death, this species is named in honor to Willi Rickmer Rickmers for his contributions to our knowledge about the Alai and Pamir regions and his outstanding work as organizer of the first German-Russian expedition to this area.
Discussion
Two studies are providing morphological data for this comparison: Gloyd and Conant (1990) and Orlov and Barabanov (1999). Both studies are strongly differing in their taxonomic concepts and therefore as well in the morphological characters provided for the different taxa. As the previous one provides more details of more taxa, the specimens from Alai were mainly compared with the data provided by Gloyd and Conant (1990), but also with the few characters given from Orlov and Barabanov (1999). The examination and comparison with both studies resulted in differences to all Gloydius taxa, but especially to both G. h. caraganus and G. h. halys. Consequently, the specimens from Alai were hence described as a new species. It was Orlov and Barabanov (1999) who already mentioned that G. h. halys and G. h. caraganus are sympatric in some areas of their distribution range. We agree with this, and think that the morphological differences are strong enough to recognize G. h. caraganus as full species. However, a further integrative revision is in need to clarify the status of the other involved taxa.
According to the unresolved systematics of the G. halys/intermedius complex the actual distribution of Gloydius in the Kyrgyz Republic is varying between the different authors and needs to be reviewed. Although several authors mention Gloydius halys as not rare in Kyrgyzstan (e.g., Yakovleva, 1964; Terent’ev and Chernov, 1965; Eremchenko et al., 1992). Moreover, Yakovleva (1964) mentioned that among the, at her time, four known subspecies, only G. h. caraganus occurs in Kirgizia and stated that the taxon has been found in almost all mountain foothills and also penetrated into mountain ranges. However, Gloyd and Conant (1990), differentiating both groups by the presence or absence of apical pits, mentioned that some of Yakovleva’s (1964) specimens could be misidentified G. intermedius and consequently recognized G. h. caraganus and G. intermedius from Kyrgyzstan. Later, Orlov and Barabanov (1999, map 1), not recognizing apical pits as a character to distinguish both groups, showed both G. h. halys and G. h. caraganus on a rough distribution map including Kyrgyzstan, but failed to present point localities or the respective material. These authors restricted G. intermedius to NE China, extreme SE Russia and the Korean Peninsula. Very recently, Sindaco et al. (2013) provided a rough scale distribution map with point localities, but only referring to G. halys as species and excluding G. intermedius. A summary of these localities referring to the different taxonomic concepts is given in fig. 5.
The different populations of the polytypic G. halys are distributed over a large area from western to eastern Central Asia. However, only three taxa, G. h. halys, G. h. caraganus and G. h. caucasicus are distributed over larger areas. Gloydius h. halys and G. h. caraganus seem to occur in Kirghizia, while other taxa are geographically restricted. Especially G. h. boehmei and G. h. caucasicus can be found in reasonable geographic distance to the Alai, and are separated from this valley by high mountain ranges like the Pamir and the Hindu Kush. In the Kyrgyz Republic Gloydius occurs within an a vertical range between 550 m in the “Chuiskaya Dolina [= valley]” (Yakovleva, 1964) and at least 4000 m on the southeastern slopes of the Ferganskiy Mountains (Chernov, 1959). Although, Gloydius rickmersi sp. n. seems to be a today Alai endemic and isolated high alpine relict population from a formerly wider distributed species. Although Gloydius taxa are very variable in their elevational distribution (e.g., G. h. halys: 150 to 2300 m; G. h. caucasicus: sea level to at least 3000 m; G. h. caraganus: below sea level to 4000 m fide Gloyd and Conant [1990]; to 1000 m fide Orlov and Barabanov [1999]) an endemic alpine species in the Alai is not surprising as it is known as an alpine biodiversity hotspot with even several endemic vertebrate taxa (e.g., Ellobius alaicus, Rodentia).
Acknowledgements
This research was financially supported by the National Geographic Society (grant no. GEFNE81-13). We are grateful to the National Academy of Sciences of Kyrgyz Republic and the Academy of Sciences of the Republic of Tajikistan for the support of the scientific expedition. We thank Spartak Litvinchuk for providing a paratype. We are grateful to Frank Tillack for his useful comments on the manuscript and to Roberto Sindaco for providing additional distribution data. PW, AT, GM and LB are grateful to all members of the Alai-Pamir expedition for the great time in the field and especially to Camilla Hansen for her help and her support of the expedition.
References
Agakhanyantz O.E., Lopatin I.K. (1978): Main characteristics of the ecosystems of the Pamirs, USSR. Arctic and Alpine Research 10: 397-407.
Ananjeva N.B., Munkhbayar K., Orlov N.L., Orlova V.F., Semenov D.V., Terbish K. (1997): Amphibians and reptiles of Mongolia. Reptiles. KMK Scientific Press Ltd., Moscow, Russia, 416 pp. [in Russian with English summary].
Ananjeva N.B., Borkin L.J., Darevsky I.S., Orlov N.L. (1998): Amphibians and reptiles. Encyclopedia of Russia’s Nature. ABF Publishing Company, Moscow, Russia, 524 pp. [in Russian].
Arevalo E., Davis S.K., Sites J.W. (1994): Mitochondrial DNA sequence divergence and phylogenetic relationships among eight chromosome races of the Sceloporus grammicus complex (Phrynosomatidae): in central Mexico. Systematic Biology 43: 387-418.
Bandelt H.-J., Forster P., Röhl A. (1999): Median-joining networks for inferring intraspecific phylogenies. Molecular Biology and Evolution 16: 37-48.
Bour R. (1993): Les voyages de Peter Simon Pallas et l’origine de Coluber halys (Serpentes, Viperidae). Bulletin Mensuel de la Société Linnéenne de Lyon 62: 328-340.
Chernov S.A. (1934): On the systematics and distribution of Agkistrodon (Ophidia) in the Soviet Union. Doklady Akademii Nauk SSSR, new series 1 (6): 350-356 [parallelly in Russian and English].
Chernov S.A. (1959): Fauna of Tadjik S.S.R. T.18. Reptiles. Academy of Sciences Tadjik S.S.R. Press, Stalinabad, 202 pp. [in Russian].
Conservation International Foundation (2014): Biodiversity hotsposts – mountains of Central Asia. Download from: www.conservation.org. Accessed on 19/05/2014.
Eremchenko V.K. (2007): Report. Bishkek. 18 pp.
Eremchenko V.K., Panfilov A.M., Tsarinenko E.I. (1992): Catalogue of the collection of amphibians and reptiles of the Zoological Museum of the Institute of Biology of the Academy of Sciences of the Republics of Kyrgyzstan. In: Konspect issledovanii po tsitogenetike i sistematike nekotorykh asiatskikh vidov Scincidae and Lacertidae [Research conspectus on cytogenetics and systematics of some Asian species of the Scincidae and Lacertidae], p. 91-176. Eremchenko V.K., Panfilov A.M., Tsarinenko E.I., Eds, Ilim, Bishkek [in Russian].
Gloyd H.K., Conant R. (1990): Snakes of the Agkistrodon Complex. A Monographic Review. Contributions to Herpetology, No. 6. Society for the Study of Amphibians and Reptiles, Oxford, OH, vi + 614 pp.; 52 pl.
Gumprecht A., Tillack F., Orlov N.L., Captain A., Ryabov S. (2004): Asian Pitvipers, 1st Edition. GeitjeBooks, Berlin, 368 pp.
Hall T.A. (1999): BioEdit: a user-friendly biological sequence alignment editor and analysis program for Windows 95/98/NT. Nucleic Acids Symposium Series 41: 95-98.
Hoser R. (2013): A review and rearrangement of pitviper genera (Serpentes: Viperidae: Crotalinae). Australasian Journal of Herpetology 19: 43-63.
Kaiser H., Crother B.I., Kelly C.M.R., Luiselli L., O’Shea M., Ota H., Passos P., Schleip W.D., Wüster W. (2013): Best practices: in the 21st century, taxonomic decisions in herpetology are acceptable only when supported by a body of evidence and published via peer-review. Herpetological Review 44: 8-23.
Karamhudoeva M. (2008): Survey of invertebrate animals of the Pamir-Alai’s Transboundary Conservation Area. Download from patca.zerofive.co.uk. Accessed on 20/12/2011.
Karpenko V.P. (1957): On distribution and ecology of the Pallas pitviper. In: Sbornik Nauchnykh Rabot (Scientific Collected Papers), Trudy Stalinabad Medical Institute, Vol. 23, p. 109-120. Koz Y.L., Ed., Stalinabad (now Dushanbe), (in Russian).
Mani M.S. (2007): Ecology and Biogeography of High Altitude Insects. Series Entomologica, Vol. 4. Springer, Dordrecht.
Orlov N.L., Barabanov A.V. (1999): Analysis of nomenclature, classification, and distribution of the Agkistrodon halys-Agkistrodon intermedius complexes: a critical review. Russian Journal of Herpetology 6: 167-192.
Orlov N.L., Barabanov A.V. (2000): About type localities for some species of the genus Gloydius Hoge et Romano-Hoge, 1981 (Crotalinae: Viperidae: Serpentes). Russian Journal of Herpetology 7: 159-160.
Orlov N.L., Sundukov Y.N., Kropachev I.I. (2014): Distribution of pitvipers of “Gloydius blomhoffii” complex in Russia with the first records of Gloydius blomhoffii blomhoffii at Kunashir Island (Kuril Archipelago, Russian Far East). Russian Journal of Herpetology 21: 169-178.
Reinig W.F. (1928): Wissenschaftliche Ergebnisse der Alai-Pamir Expedition. Beiträge zur Faunistik des Pamir-Gebietes. Berlin.
Reinig W.F. (1932): Wissenschaftliche Ergebnisse der Alai-Pamir Expedition, Teil III. Beiträge zur Faunistik und Ökologie des Pamir-Gebietes, Band I: Ökologie und Tiergeographie. Berlin.
Rickmer Rickmers W. (1929): Alai-(Pamir-)Expedition 1928 (Vorläufige Berichte der deutschen Teilnehmer): Aus der Arbeit der Notgemeinschaft der Deutschen Wissenschaft, Heft 10. K. Siegesmund Verlag, Berlin.
Rickmer Rickmers W. (1930): Alai! Alai! Arbeiten und Erlebnisse der Deutsch-Russischen Alai-Pamir-Expedition. Brockhaus-Verlag, Leipzig.
Said-Aliev S.A. (1979): Amphibians and Reptiles of Tajikistan. Donish, Dushabe [in Russian].
Sambrook J., Fritsch E.F., Maniatis T. (1989): Molecular Cloning: A Laboratory Manual. Cold Spring Harbor Laboratory Press, New York.
Sindaco R., Venchi A., Grieco C. (2013): The Reptiles of the Western Palearctic. 2. Annotated Checklist and Distributional Atlas of the Snakes of Europe, North Africa, Middle East and Central Asia, With an Update to the Vol. 1. Monografie della Societas Herpetologica Italica II. Edizioni Belvedere, Latina, Italy, 543 pp.
Tamura K., Stecher G., Peterson D., Filipski A., Kumar S. (2013): MEGA6: Molecular Evolutionary Genetics Analysis version 6.0. Molecular Biology and Evolution 30: 2725-2729.
Terent’ev P.V., Chernov S.A. (1965): Key to the Amphibians and Reptiles. Translated from Russian [1949]. Israel Program for Scientific Translations, 315 pp.
Wagner P., Bauer A.M., Wilms T.M., Leviton A., Tillack F., Böhme W. (in rev.): Exploring herpetodiversity using biodiversity archives – a checklist of the amphibians and reptiles of Afghanistan.
Xu Y., Liu Q., Myers E.A., Wang L., Huang S., He Y., Peng P., Guo P. (2012): Molecular phylogeny of the genus Gloydius (Serpentes: Crotalinae). Asian Herpetological Research 3: 127-132.
Yakovleva I.D. (1964): Reptiles of Kirgizia. Ylym, Frunze, Khirgiz, S.S.R., 272 pp. [in Russian].
Appendix. Material examined
G. h. boehmei. Afghanistan. ZFMK 8648, Andarab valley.
G. h. caraganus. Kazakhstan. ZFMK 16385, Tscheskaskan. Kirghizia. ZFMK 44983, Frunse.
G. h. caucasicus. Iran. ZFMK 89231-33, Laar valley.
G. h. halys. Kazakhstan. ZFMK 59366, without further locality. Mongolia. ZFMK 59365, Chuzhird.
“G. intermedius”. Mongolia. ZFMK 44228 Gobi, Dalan. Russia. ZFMK 51068-69, Buchta Mekovodnaja; ZFMK 44984-85, Tschingan; ZFMK 49054-56, Ussuri NP.
Footnotes
Associate Editor: Sylvain Ursenbacher.
