Zootaxa 3636 (3): 401–420 ISSN 1175-5326 (print edition) www.mapress.com/zootaxa/ Article ZOOTAXA Copyright © 2013 Magnolia Press ISSN 1175-5334 (online edition) http://dx.doi.org/10.11646/zootaxa.3636.3.1 http://zoobank.org/urn:lsid:zoobank.org:pub:13E09793-A6C9-4CF2-9ED8-55A2A9F04541 A new species of Bachia Gray, 1845 (Squamata: Gymnophthalmidae) from the western Brazilian Amazonia MAURO TEIXEIRA JR1,2, FRANCISCO DAL VECHIO1, PEDRO M. SALES NUNES1, ANTONIO MOLLO NETO3, LUCIANA MOREIRA LOBO4, LUIS FERNANDO STORTI5, RENATO AUGUSTO JUNQUEIRA GAIGA6, PEDRO HENRIQUE FREIRE DIAS6, MIGUEL TREFAUT RODRIGUES1 1Laboratório de Herpetologia, Departamento de Zoologia, Instituto de Biociências, Universidade de São Paulo, CEP 05508-090, São Paulo, SP, Brazil. 2E-mail: [email protected] 3Universidade Federal do ABC, Centro de Ciências Naturais e Humanas, Laboratório de Vertebrados. Av. dos Estados 5001, Bloco A, 6° andar DI624. CEP 09210-971. Santo André, SP, Brazil. 4Alimini Consultoria Cientifica LTDA, R. Manoel Vieira Sarmento 03, Chácara Santana, CEP 05831-150, São Paulo, SP, Brazil. 5Departamento de Biologia Animal e Vegetal, Museu de Zoologia, Universidade Estadual de Londrina, CEP 86051-990, Londrina, PR, Brazil. 6Biotropica Consultoria Ambiental LTDA, Av. Santo Antônio 571, Jd. Cascatinha, CEP 37701-036, Poços de Caldas, MG, Brazil. Abstract A new species of Bachia of the B. dorbignyi group, Bachia scaea sp. nov., is described from the left bank of the upper Madeira River, at Rondônia state, at the western Brazilian Amazonia. The new species resembles morphologically B. dor- bignyi and B. peruana, and seems to be related with the former species based on molecular data (16S and c-mos sequenc- es). Nonetheless the presence of a first temporal separating parietal and supralabial scales and the absence of clawed fingers in the new species, can promptly distinguish it from their close relatives. This description ends with several-de- cades of stasis in the taxonomy of the Bachia dorbignyi group from Amazonian lowlands, and also presents new evidence that supports the Madeira River as a vicariant barrier. Key words: Bachia scaea sp. nov., fossorial habits, Amazon Forest Introduction Bachia species of the B. dorbignyi group are widely distributed over the western Amazon Forest and the Andean slopes (Dixon 1973). The first described species was the in this group originally placed in the genus Chalcides by Duméril and Bibron (1839) (C. dorbignyi, from Santa Cruz, Bolivia). Later, Gray (1845) recognized Duméril and Bibron’s species as belonging to a distinct genus, and described Bachia to accommodate it. By the end of the XIX century, Cope (1868; 1896) described Heteroclonium bicolor and Ophiognomon trisanale, both now in the genus Bachia (Dixon 1973). During the first decades of the following century four more species were described, by Werner (1901), in the genus Cophias (C. peruanus), and also by Noble (1920) (B. intermedia), Ruthven (1925) (B. talpa), and Burt and Burt (1931) (B. barbouri). Finally, Dixon (1973) in his revision of the genus Bachia, described the last species, B. huallagana, and the B. dorbignyi group achieved its current content. Although other Bachia groups, such as the B. bresslaui one, have experienced a high number of descriptions in recent years (Castrillon & Strussmann 1998; Kizirian & McDiarmid 1998; Rodrigues et al. 2007, 2008; Freitas et al. 2011), the B. dorbignyi group has witnessed a long taxonomic stasis. This traditional arrangement of Bachia in species groups based on morphological features, as defined by Dixon (1973), has been recently challenged, as molecular phylogenetic approaches are showing that they may not represent natural arrangements, as they are not monophyletic (Kohlsdorf & Wagner 2006; Galis et al. 2010; Kohlsdorf et al. 2010). The species from B. dorbignyi group appear in distinct clades along the topology in different molecular studies, however as for the other groups, the monophyly is never recovered (Kohlsdorf & Accepted by S. Carranza: 4 Mar. 2013; published: 5 Apr. 2013 401 Wagner 2006; Galis et al. 2010; Kohlsdorf et al. 2010). The topologies themselves are also not in agreement, and further studies need to be done to solve this question. Herein based on specimens from the left bank of upper Madeira River we recognize and describe a new species of Bachia, which can be assigned morphologically to B. dorbignyi group, along with information on its natural history, and a hypothesis of its phylogenetic position. Material and methods Sampling methods The field samplings were carried out during a three years period (2010–2012), with four surveys of 15 days every year (January, April, July and October). Three main areas (general coordinates: [1] 9°26' S, 64°49' W; [2] 9°35' S, 65° 3' W, and [3] 9°36' S, 65°24' W) were inventoried, at both banks of Madeira River, along Porto Velho municipality, Rondônia state, Brazil. Active samplings were performed by 12 persons each sampling period, and were complemented with pitfall traps; each trap consisting of seven aligned 100L buckets buried at ground level, connected with a 10m long and 50 cm high plastic vertical fence. The total effort was of 9,213 hours/man in the active search and 5,380 buckets/day. Additionally, a few more specimens found at the vicinities of Porto Velho city town were also included in our type series. Morphology All morphological analyses were made after fixation. Length measurements were taken to the nearest 0.1mm with a Mitutoyo digital caliper; scale counts and the analysis of other morphological characters were performed with a stereomicroscope Zeiss STEMI SV6. Scale nomenclature follows Dixon (1973). Although the grouping arrangement proposed by Dixon (1973) may not represent monophyletic groups (Kohlsdorf & Wagner 2006; Galis et al. 2010; Kohlsdorf et al. 2010), the new species shows morphological features that fit the characterization of what Dixon (1973) defined as the B. dorbignyi group; thus for comparative purposes we compared the new species with the other groups as a whole, and with those within the B. dorbignyi group in more detail. Comparisons were made with specimens housed in the herpetological collection of the Museu de Zoologia da Universidade de São Paulo (MZUSP), Universidade Federal do Acre (UFAC) and Museu de Zoologia da Universidade Estadual de Campinas (ZUEC) (Appendix I), and with data from the literature for both external (Vanzolini 1961a,b; Dixon 1973; Avila-Pires 1995; Castrillon & Strussmann 1998) and hemipenial morphology (Cope 1896; Presch 1978; Nunes 2011). Meristic characters were: (SC) superciliaries; (TS) temporal scales; (FP) femoral pores; (PP) preanal pores; (PS) preanal shields; (SAB) scales around midbody; (DO) dorsals scales counted from parietals to the row over cloacal region; (VE) ventrals scales counted between interbrachials and preanal shields; (GU) gular scales, between mental plates and interbrachials ; (CA) caudal scales. The snout-vent length (SVL) and tail length (TL) were also measured, the last one only on intact tails. A hemipenis of one individual of the new species, and one of B. dorbignyi were prepared following the procedures described by Manzani and Abe (1988), modified by Pesantes (1994) and Zaher (1999). The retractor muscle was manually separated and the everted organ filled with stained petroleum jelly. The organ was immersed in an alcoholic solution of Alizarin Red for 24 hours in order to stain eventual calcified structures (e.g. spines or spicules), an adaptation of the procedures described by Uzzell (1973) and Harvey and Embert (2008). The terminology of hemipenial structures follows Dowling and Savage (1960), Savage (1997) and Myers and Donnelly (2001, 2008). Distribution Distributional records of species of the Bachia dorbignyi group were compiled from the literature (Burt & Burt 1931; Vanzolini 1961a,b; Donosos-Barros 1968; Dixon 1973; Avila-Pires 1995; Castrillon & Strussmann 1998; Dirksen & De la Riva 1999; Silva 2005; Jerez & Tarazona 2009; Bernarde et al. 2011), online databases (GBIF 2012; SpeciesLink 2012, HerpNET [www.herpnet.org]) and from examined specimens (Appendix I). Molecular phylogeny To infer the relationship of the new species with the remaining Bachia species, we used a molecular phylogenetic 402 · Zootaxa 3636 (3) © 2013 Magnolia Press TEIXEIRA JR. ET AL. approach. The DNA was extracted from tissue samples (from liver or tail muscle) preserved in 100% ethanol (Fetzner 1999). Two mitochondrial (16S and 12S) (Palumbi 1996) and one nuclear (c-mos) (Godinho et al. 2005) fragments were amplified using standard PCR protocols. The PCR cycle protocol consisted of one initial cycle of 94°C for 5 min followed by 35 cycles of 94°C for 40 sec, 51°C for 16S and 56°C for 12S and 52°C for C-mos for 40 sec, 72°C for 40 sec and 72°C elongation for 7 min. PCR products were directly purified with Exonuclease I and Shrimp Alkaline Phosphatase (USB or Fermentas). Automated sequencing was carried out using BigDye Terminator v3.1 Cycle Sequencing kit (Applied Biosystems), followed by analysis on ABI Prism 310, 3700 or 3170 Genetic Analyzer Sequencers (Applied Biosystems) at the Instituto de Química da Universidade de São Paulo (IQUSP, São Paulo, Brazil) and Instituto de Ciências Biomédicas da Universidade de São Paulo (ICB, São Paulo, Brazil). Resulting sequences were manually edited in CodonCode Aligner v.3.7.1.1. (http://www.codoncode.com). Three individuals of the new species were included along with 15 other Bachia taxa as ingroups, and Potamites ecpleopus (Cope, 1875), Placosoma glabellum (Peters, 1870), Cercosaura eigenmanni (Griffin, 1917) and C. oshaughnessyi (Boulenger, 1885), as outgroups (Appendix II). Although we sequenced the 12S gene, we noticed that the 12S sequences available at GeneBank (DQ383203–DQ383212), are identical among several Bachia species [B. barbouri, B. heteropa alleni (Barbour, 1914), B. huallagana, B. intermedia, B. m. monodactylus (Daudin, 1802), B. m. parkeri Ruthven, 1925, B. panoplia Thomas, 1965, B. peruana, B. scolecoides Vanzolini, 196l, B. trisanale], therefore we exclude them from our analysis. Each gene was aligned separately using ClustalX v.2.