Zootaxa 3731 (3): 345–370 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.3731.3.5 http://zoobank.org/urn:lsid:zoobank.org:pub:6D082EF3-DA05-41FE-8D7D-662F800907C8

Rediscovery of the Earless Microteiid Lizard Anotosaura collaris Amaral, 1933 (Squamata: Gymnophthalmidae): A redescription complemented by osteological, hemipenial, molecular, karyological, physiological and ecological data

MIGUEL TREFAUT RODRIGUES1,3, MAURO TEIXEIRA JR1, FRANCISCO DAL VECHIO1, RENATA CECÍLIA AMARO1, CAROLINA NISA1, AGUSTÍN CAMACHO GUERRERO1, ROBERTA DAMASCENO2, JULIANA GUSSON ROSCITO1, PEDRO M. SALES NUNES1, & RENATO SOUSA RECODER1 1Departamento de Zoologia, Instituto de Biociências, Universidade de São Paulo, Caixa Postal 11.461, CEP 05508-090, São Paulo, SP, Brazil 2University of California, Berkeley, Museum of Vertebrate Zoology and Department of Integrative Biology, 3101 Valley Life Sciences Building, Berkeley, CA 94720–3140, USA 3Corresponding author. E-mail: [email protected]

Abstract

More than a century after its discovery by Ernest Garbe, and almost 80 years after its original description, we obtained a series of specimens of the earless gymnophthalmid Anotosaura collaris, the type species of the genus, up to now known only by a single specimen. On the basis of the material obtained at and close to the type locality we redescribe the species, adding information about the external and hemipenial morphology, osteology and karytoype. Molecular data confirm its sister relationship with Anotosaura vanzolinia as well as the close relationship of Anotosaura with the Ecpleopodini Colo- bosauroides and Dryadosaura. We supplement this information with thermophysiological, ecogeographical, karyotypic and ecological data.

Key words: Anotosaura collaris, Gymnophthalmidae, Brazil, earless lizard, Caatinga

Introduction

In the thirties of the last century, after a relatively long period of stasis in the taxonomic knowledge of Brazilian lizards, Afrânio do Amaral, the Director of Instituto Butantan, turned his attention to the biological collections of the Museu Paulista, latter dismembered into several institutions, one of which came to be the present Museu de Zoologia, Universidade de São Paulo (MZUSP). Although his main research focus was on uncovering undetected lizard parasites, he found several apparently undescribed lizards collected by the Museum’s naturalists in different parts of Brazil. Ernest Garbe was one of those. He was hired in 1901 and since then was actively engaged in collecting specimens from unexplored parts of Brazil and sending the samples to the Museum. Amaral eventually published an important paper on this material, in which he describes 17 new lizard species, some of which as new genera (Amaral 1933). Although several were later considered synonyms of previously known species, others remain valid and are still extremely rare. Examples are the microteiid Anotosaura collaris Amaral, 1933, only known by its type specimen, Anolis nasofrontalis Amaral, 1933 and A. pseudotigrinus Amaral, 1933, known from only a few individuals (Williams & Vanzolini 1980), all collected by Garbe. In 1908, in one of the first systematic explorations of the Brazilian Caatingas, Garbe was sent to the interior of state of Bahia, from where he returned in 1909 with an unprecedented amount of new material (Ihering & Ihering 1911). At Villa Nova da Rainha, presently municipality of Senhor do Bonfim, Garbe obtained the first, and up to now the only known, specimen of the earless microteiid lizard described by Amaral as a new genus and species, Anotosaura collaris.

Accepted by S. Carranza: 29 Sept. 2013; published: 31 Oct. 2013 345 Besides references in checklists (Amaral 1937, 1938; Ruschi 1966; Peters & Donoso-Barros 1970), Anotosaura collaris is rarely cited in the literature. In his revision of the genus Bachia, Dixon considered Anotosaura as a valid genus and suggested a close relationship with Heterodactylus and Bachia (Dixon 1973). Later on, based on new material, he reviewed Anotosaura and described a new subspecies, Anotosaura collaris vanzolinia Dixon, 1974, from Agrestina, in the state of Pernambuco (wrongly spelled Argestina), and the new species Anotosaura brachylepis Dixon, 1974, from Serra do Cipó in the State of Minas Gerais (Dixon 1974). That was a time when subspecies were widely used in Herpetology Systematics. In view of the minor differences between Amaral’s type and the specimens from Agrestina (presence versus absence of prefrontals), Dixon suggested that hybridization was possible between the two subspecies (Dixon 1974). Contrarily, the distinctiveness of A. brachylepis from its congeners was so striking that he initially hesitated in attributing it to the new genus, but ended up keeping the new species in Anotosaura (Dixon 1974). Two years later, Vanzolini (1976) reexamined the question and kept A. brachylepis in the genus, but raised A. c. vanzolinia to specific rank, a position reinforced later when discussing their distribution (Vanzolini & Ramos 1977). Since then, these three species have been associated with Anotosaura until Pellegrino et al. (2001) removed A. brachylepis from Anotosaura and proposed the new genus Rhachisaurus to accommodate it, based on molecular data. Specimens from the Atlantic Forest of northeastern Brazil that were referred to as Anotosaura sp. n. in the literature (Rodrigues 1990; Pellegrino et al. 2001) were later reallocated into the new genus Dryadosaura (Rodrigues et al. 2005). Of the two species presently recognized in Anotosaura restricted to the semiarid Brazilian Caatingas, our knowledge rely solely on Anotosaura vanzolinia, which occurs only in mesic habitats in this domain (Rodrigues 1986, 1990, 2003; Delfim & Freire 2007; Freire et al. 2009; Oliveira 2011; Gonçalves et al. 2012). In recent analyses based on both molecular and morphological data, Anotosaura vanzolinia has been systematically recovered in close association with Dryadosaura and Colobosauroides, as part of the Ecpleopodini radiation of microteiids (Pellegrino et al. 2001; Castoe et al. 2004; Rodrigues et al. 2005; Peloso et al. 2011). However the relationships as well as ecogeographical differences among these three genera remain unclear. Recently, more than a century after its discovery by Ernest Garbe, and almost 80 years after its original description by Amaral, we obtained new specimens of Anotosaura collaris from the type locality and nearby areas. This new material allowed us to redescribe this earless elongated fossorial species, detailing its external and hemipenial morphology and osteology. Additionally, in order to test the generic allocation of A. vanzolinia and the relationship of A. collaris with Dryadosaura and Colobosauroides we obtained a phylogeny based on mitochondrial and nuclear genes. Finally, we complemented our analysis with a set of ecological, thermophysiological, ecogeographical and karyotype data.

Material and methods

Field work. We carried out a herpetofaunal survey at the northern portions of the relatively isolated Jacobina mountain range at Chapada Diamantina, a subsection of the Espinhaço range, in northern Bahia. We visited the localities: Senhor do Bonfim (Serra da Maravilha, Coqueiral, and Alto da Rainha mountains, and Missão do Sahy village), Campo Formoso (Serra do Cruzeiro mountain), Antônio Gonçalves (Serra da Gameleira and Serra do Sobradinho mountains), Pindobaçu (Serra da Paciência mountain) and Jaguarari (Beringela and Catuni villages). Specimens were collected using pitfall traps and through active search. The pitfall traps were installed at forest and savanna sites close to Campo Formoso and Antonio Gonçalves towns, covering altitudes between 790 and 980 m a.s.l. Each trap consisted of four 30 L buckets buried on the ground, one central and three radials, connected to each other with a 4 m long and 50 cm high plastic fence. Twenty-five traps were installed and remained opened from 9th December 2012 to 3rd January 2013, totalizing an effort equivalent to 2,500 buckets\day. Active search was performed at the other localities, mainly in Caatinga and campos rupestres (rocky meadows) habitats. Nearly every day, active search sessions were typically performed by 6–7 persons for about 2h. The sessions took place during the morning, early evening, and at the beginning of the night, and involved searching through a diversity of microhabitats at each locality. Thermal physiology and environmental data. We estimated the critical thermal maximum (CTmax) and minimum (CTmin) of six individuals from Senhor do Bonfim and Campo Formoso, right after capture. CTmax and CTmin are indices that represent the temperatures at which locomotory response is compromised yet the lizards

346 · Zootaxa 3731 (3) © 2013 Magnolia Press RODRIGUES ET AL. will recover (Cowles & Bogert 1944). They were measured by heating up or cooling down the lizards until righting response was lost (Brooks & Sassman 1965). For each experiment, a single animal was placed in a deli cup where a thermocouple (type-T) was attached so that its tip was upright inside the cup. The deli cup was placed close to a heating pad (for CTmax) or inside a mini freezer (for CTmin) and righting response was tested by gently displacing the lizard from its normal resting position, flapping the deli cup so the animal would rest in its back and touch the thermocouple. Rate of temperature change was kept around 0.5–0.75 oC/minute, during experiments. After reaching a given CT, the animal was brought back to a comfortable temperature. There was an interval of at least 18 hours in between experiments with the same individual. To measure variation in environmental temperatures during the warmest part of the year, we deployed 4 dataloggers (each equipped with two silvered cylindrical aluminum probes of 3x0.3 cm) that collected temperature every 10 minutes for 18 hours (11th–12th December 2012), in a mountain close to our field base, within the altitude range where Anotosaura collaris specimens were found. Two dataloggers were placed in a forested area (10°33'45.51"S, 40°24'36.45"W, 979 m a.s.l.), one measuring temperatures on the surface of the leaf litter and the other under the litter. Two other dataloggers were deployed in a nearby rocky open habitat (10°35'26.76"S, 40°23'24.87"W, 773 m a.s.l.), in which we measured the temperature both on the surface and underneath the rocks. We explored ecogeographical traits that could differentiate Anotosaura collaris from the close related Ecpleopodini Colobosauroides cearensis Cunha, Lima-Verde & Lima, 1991, Dryadosaura nordestina Rodrigues, Freire, Pellegrino & Sites, 2005, and A. vanzolinia. We thus gathered locality records for all four species from the literature (Amaral 1933; Dixon 1973, 1974; Vanzolini 1976; Soares & Caramaschi 1998; Borges-Nojosa & Caramaschi 2003; Rodrigues et al. 2005; Delfim & Freire 2007; Falcão & Hernández 2007; Camacho & Rodrigues 2006; Santana et al. 2008; Freitas & Moisés 2009; Gogliath et al. 2010; Noronha-Oliveira et al. 2010; Moura et al. 2011; Delfino & Soeiro 2012; Gonçalves et al. 2012), the records from the Museu de Zoologia da Universidade de São Paulo (MZUSP), and our field work. Then, using global climatic (Worldclim v. 1.4) (Hijmans et al. 2005) and soil (HWSD v. 1.2) (Nachtergaele et al. 2012) databases (at the highest resolution available for soil layers—arc of 5 min), we extracted information from the several variables. Soil texture as measured as the proportion of clay, silt and sand, was coded according to the United States Department of Agriculture texture classes (Nachtergaele et al. 2012), with values greater than 10 indicating sandy soils and those below 5, indicating clayish soils. We also extracted the following bioclim variables using the R packages “raster” (Hijmans & van Etten 2013) and “MASS” (Venables & Ripley 2002): annual precipitation (BIO12), precipitation seasonality (BIO15), maximum temperature of the warmest month (BIO5), minimum temperature of the coldest month (BIO6), daily temperature range (BIO2), and temperature seasonality (BIO4). Morphometry. Twelve measurements were taken with a digital caliper (to nearest 0.01 mm) from preserved specimens: snout-vent length (SVL), from border of cloaca to the tip of snout; length between limbs (LBL), between the anterior margin of hindlimb to the posterior margin of forelimb; body width (BW), anteriorly to hindlimb insertion; head height (HH), at highest point in the longitudinal axis; head width (HW), at its widest point; head length (HL), from the posterior margin of tympanic aperture (or post-temporal collar groove in earless species) to the tip of snout; femur length (FEM), from the knee joint to margin of outer scale of anal plate; tibia length (TIB), from the knee joint to margin of sole; hind foot length (FTL), from margin of sole to tip of fourth toe, excluding claw; humeral length (HUM), from axilla to elbow joint; forearm length (FAL), from elbow joint to tip of fourth finger, excluding claw; forearm width (FAW), at its widest point. Limb measurements were taken on the right side. Additionally, tail length (TAL) was measured in individuals that had intact tails (regenerated tails were distinguished by the presence of darker and narrower scales).

Measurements were log10 transformed prior to analysis and were tested for normality with Shapiro Wilk’s Test and equivalence of variances with Levene’s Test (Zar 2010). Two-way analysis of variance (ANOVA) was used to test for differences between sexes of Anotosaura and between A. collaris and A. vanzolinia. When normality or equivalence of variances was not met, differences were tested with the non-parametric Kruskal-Wallis Test. To assess the pattern of morphological differentiation among Anotosaura spp. and the related Colobosauroides cearensis and Dryadosaura nordestina (Rodrigues et al. 2005, see molecular results) we performed a discriminant analysis using log-transformed morphometric data (excluding TAL because of missing data). Only males were used in this analysis to avoid effects of sexual dimorphism and sampling bias in among-group variation. All statistical analysis were performed using IBM SPSS Statistics (version 20.0, SPSS Inc., 2011). The individuals examined are housed at the herpetological collection of MZUSP and the Laboratório de

REDISCOVERY OF ANOTOSAURA COLLARIS Zootaxa 3731 (3) © 2013 Magnolia Press · 347 Herpetologia, Instituto de Biociências da Universidade de São Paulo (MTR) (Appendix I). Additional information on morphological characters for comparisons was obtained from the literature (Dixon 1974; Cunha et al. 1991; Soares & Caramaschi 1998; Rodrigues et al. 2005). Hemipenial morphology. The hemipenis of one individual of Anotosaura collaris (MZUSP 103845) and one of A. vanzolinia (MZUSP 95328) 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), in an adaptation proposed by Nunes et al. (2012) 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 Donnely (2001, 2008). Osteology. One individual of Anotosaura collaris (MZUSP 103837) was scanned with a SkyScan digital microtomography (www.skyscan.be), with a resolution of 9 µm and no filters added. The controlling software was the NRecon Server v. 1.6. The images were processed with CT Analyzer v. 1.11 software, setting an automatic threshold point, at the skull level, for filtering low-density matter (soft tissue). The resulting 3D model of the skeleton was then used to recover the osteological data, visualized on CT Volume v. 2.2. The anatomical terminology follows Romer (1956), Hoffstetter and Gasc (1969), Bell et al. (2003), Evans (2008), Russel and Bauer (2008), and Jerez and Tarazona (2009). Cytogenetics. Cytogenetic analyses were carried out on four specimens (MZUSP 103840, MZUSP 104210, MZUSP 103832, MZUSP 103833). Mitotic metaphases were obtained either from suspension of liver cells or intestine after in vivo colchicine treatment, according to conventional protocols (Yonenaga 1972; Bogart 1973). Chromosome studies were performed after Giemsa staining. Molecular phylogeny. We used a molecular phylogenetic approach to infer the taxonomic status of Anotosaura collaris and its relationships with the closely related Ecpleopodini genera Dryadosaura, Colobosauroides, Arthrosaura, Marinussaurus, Leposoma and Ecpleopus (Pellegrino et al. 2001; Rodrigues et al. 2005; Peloso et al. 2011). Three individuals of A. collaris, one A. vanzolinia, and one of Dryadosaura nordestina were sequenced for this study (Appendix II). Bachia dorbignyi (Duméril & Bibron, 1839), Cercosaura ocellata ocellata Wagler, 1830, and Potamites ecpleopus (Cope, 1875) were used as outgroups (Appendix II). One sequence (18S) available on GenBank, from Marinussaurus (Peloso et al. 2011) was very divergent, probably due to contamination, and was therefore not used in this study. The DNA was extracted from tissue samples (liver or tail muscle) preserved in 100% ethanol (Fetzner 1999). For the molecular analyses, we used three mitochondrial markers, ribosomal RNA 12S and 16S (Palumbi, 1996) and the protein-coding ND4 (Arévalo et al. 1994); one nuclear protein-coding gene, C-mos (Godinho et al. 2005) and one nuclear ribosomal RNA 18S (Pellegrino et al. 2001). All genes fragments were amplified using standard PCR protocols, with annealing temperatures of 49°C for ND4; 51°C for 16S; 56°C for 12S; 52°C for C-mos; and 54°C for 18S. PCR products were sequenced at Instituto de Química da Universidade de São Paulo and Instituto de Ciências Biomédicas da Universidade de São Paulo. Resulting sequences were manually edited in CodonCode Aligner v. 3.7.1.1. (http://www.codoncode.com), and aligned using MUSCLE program (Edgar 2004), embedded in MEGA v. 5 software, in its default settings (Tamura et al. 2011). The protein coding genes ND4 and C-mos were translated into amino acids to check the alignments. The best-fit model of evolution for each alignment was tested using jModelTest v. 2.1.3 software (Posada 2008) and Akaike Information Criterion (AIC) (locus 12S—model GTR+G+I; locus 16S—model GTR+G; locus ND4—model HKY+G+I; locus C-mos—model HKY+G; and locus 18S—model JC). The concatenated alignment consisted of a molecular dataset of 2,524 bp: 12S = 439 bp (171 variable sites); 16S = 530 bp (149 variable sites); ND4 = 736 bp (449 variable sites); c-mos = 406 bp (163 variable sites); 18S = 413 bp (7 variable sites). We inferred phylogenetic relationships using Bayesian and Maximum Likelihood methods on a concatenated dataset. The Bayesian analysis (BA) was performed with MrBayes v. 3.2 (Ronquist et al. 2012), available on Cipres Science Gateway (Miller et al. 2010) including five partitions according to the evolution model selected. Two independent Bayesian runs were performed with four Markov chains, starting with a random tree. Each run consisted for 10,000,000 generations with trees being sampled each 1,000 generations. We discarded 25% of the initial trees as burnin, by inspecting log-likelihood traces using Tracer v. 1.5 (Rambaut & Drummond 2009). The convergence for independent runs and ESSs were checked with Tracer v. 1.5 (Rambaut & Drummond 2009). Maximum likelihood (ML) bootstrapping (1,000 replicates) was performed in RAxML 7.4.4. (Stamatakis 2006) also available on Cipres

348 · Zootaxa 3731 (3) © 2013 Magnolia Press RODRIGUES ET AL. Science Gateway (Miller et al. 2010). The majority 50% consensus trees saved with posterior probabilities and bootstrap values on the nodes were visualized using FigTree 1.3.1 (http://tree.bio.ed.ac.uk/). Uncorrected genetic distances (p-distances) were calculated using PAUP* v 4.0b10 (Swofford 2001).

Results

Taxon redescription

Anotosaura collaris Amaral 1933 (Figs. 1–3A,B, 4A, 5)

Anotosaura collaris Amaral 1933: 69; Anotosaura collaris—Dixon 1973: 8; Anotosaura collaris collaris— Dixon 1974: 14; Anotosaura collaris— Vanzolini 1976: 120.

Holotype. MZUSP 788. Type Locality: “Villa Nova, Bahia, Brasil” Additional material. 16 specimens (8 females, 8 males): MZUSP 103834 (field number MTR 24561), MZUSP 103836–103840 (MTR 24563–24567), MZUSP 104210, (MTR 24568), MZUSP 103841–103844 (MTR 24805–24808), MZUSP 103845 (MTR 25010), MZUSP 103846 (MTR 25022): Alto da Rainha: Senhor do Bonfim, Bahia (10°26'24.56"S, 40°10'32.19"W, 715m a.s.l.), 13–31th December 2012. MZUSP 103835 (MTR 24562): Missão do Sahy: Antonio Gonçalves, Bahia (10°31'3.66"S, 40°14'54.48"W, 895m a.s.l.), 13th December 2012. MZUSP 103832, 103833 (MTR 24620, 24621): Morro do Cruzeiro: Campo Formoso: Bahia (10°30'56.12"S, 40°18'31.51"W, 779m a.s.l.), 15th December 2012. All specimens collected by the authors. External morphology. Rostral broad, wider than high, contacting first supralabial, nasal and frontonasal. Frontonasal heptagonal, slightly wider than long, contacting rostral, nasal, loreal and prefrontals. Prefrontals hexagonal, as long as wide, in broad contact at midline. Frontal heptagonal, twice longer than wide; anterior margin angulose, indenting prefrontals, lateral margins almost parallel, slightly divergent posteriorly, contacting broadly the second and less the third supraocular; posterior margins diagonally contacting parietals and in straight contact with interparietal. Frontoparietals absent. Interparietal rectangular, longer than wide, as long as and narrower than frontal, narrower than parietals. Lateral margins of interparietal slightly concave. A pair of very large irregularly hexagonal parietals in straight lateral contact with interparietal, anteriorly contacting frontal and laterally in broad contact with third supraocular. External part of parietals slightly rounded, contacting two large temporal and two smaller occipital scales; in some specimens parietal contact sixth supralabial. Three supraoculars, first the smallest, second the largest, as large as prefrontal; first supraocular contacting only prefrontal, loreal, and second supraocular, separated by the frontal by the contact between second supraocular and frontal; third supraocular in broad contact with parietal and an enlarged postocular. Nasal above first and second supralabials, and also contacting loreal, frontonasal and rostral; large, longer than high, with nostril centrally placed in the lower part of scale. Loreal posterior to nasal, narrower and diagonally oriented; contacting frontonasal, prefrontal, first supraocular, first supraciliar, the frenocular, and second supralabial. Frenocular small, quadrangular, followed posteriorly by two to four suboculars, the second being the largest and located under the eye. Six supralabials, second and third subequal and smallest, fourth under the eye, sixth the highest and the longest, fifth supralabial separated from parietal by an enlarged postocular. Sixth supralabial followed posteriorly by a minute and elongate granule which contacts granules in the ear depression. Two to four superciliaries (usually two), the first being slightly longer, their suture indented by second supraocular. First superciliar contacting loreal, first and second supraoculars, frenocular, second superciliary, and an elongated, almost granular, preocular. Central part of eyelid with a single semitransparent disc surrounded by small, slightly pigmented, and irregularly shaped smooth granules. Lower eyelid with seven to nine strongly pigmented palpebrals. Temporal region with four large, smooth and juxtaposed scales between parietal, sixth supralabial, and the ear; two of them diagonally contacting the sixth labial. First temporal contacting parietal, second and third temporals, fifth and sixth supralabials and postocular. External auditory meatus absent, tympanum indistinct. Region of ear opening situated in the middle of a vertical incomplete fold covered by a series of small and juxtaposed granules extending to the gular region. Granules corresponding to the ear area contacting anteriorly two large temporals, and two much smaller, elongated, and

REDISCOVERY OF ANOTOSAURA COLLARIS Zootaxa 3731 (3) © 2013 Magnolia Press · 349 juxtaposed flat granules. Lateral surface of neck with smooth scales and occasionally granules, irregular in size and shape, varying from juxtaposed to slightly imbricate, and arranged in regular transverse series between ear and shoulder. All head scales smooth and juxtaposed.

FIGURE 1. Lateral (A), ventral (B) and dorsal (C) views of the head, and (D) of the entire body, in ventral (above) and dorsal (below) views of the holotype of Anotosaura collaris (MZUSP 788). Scale bar = 1mm.

350 · Zootaxa 3731 (3) © 2013 Magnolia Press RODRIGUES ET AL. FIGURE 2. Lateral (A), ventral (B) and dorsal (C) views of the head, ventral views of right hand (D) and foot (E), and the cloacal region (F) of Anotosaura collaris (MZUSP 103832). Scale bars = 1 mm.

Mental broad, wider than high. Postmental heptagonal, wider than long. Three pairs of genials, all contacting infralabials; first the smallest, third the largest, first and second in broad contact at midline; third separated at midline by a small granule and deeply indented by a pair of flat, enlarged, and diagonally disposed scutes. Five infralabials, first the smallest, all others subequal. Gulars smooth, imbricate, quadrangular, laterally juxtaposed, irregular in size, in five regular transverse rows; third and fifth rows with the longest scales. Gulars followed by a distinct interbrachial region with eight larger and elongate scales. A distinct collar fold characterized by some granules and reduced scales in the second row of gulars preceding the interbrachial row, another one between interbrachials and the last row of gulars. Dorsal scales disposed in regular transversal rows, anteriorly smooth, imbricate, rounded in the occipital region, becoming progressively narrower, more elongate and rectangular towards the arm level, being progressively hexagonal with lateral sides almost juxtaposed posteriorly. Twenty-eight or 29 transverse rows of dorsals between interparietal and the posterior level of the hindlimbs. Lateral scales about the same size as dorsals, rectangular, imbricate laterally, not acuminate and more diagonally oriented than dorsals; those closer to ventrals, the larger. An irregular series of transversally arranged scattered granules in the skin separates transverse series of lateral scales. A distinct area with granular scales surrounds the area of arm insertion. Twenty three to 25 scales

REDISCOVERY OF ANOTOSAURA COLLARIS Zootaxa 3731 (3) © 2013 Magnolia Press · 351 around midbody. Ventral scales smooth, laterally juxtaposed, slightly imbricate antero-posteriorly, rectangular, and rounded posteriorly; first rows almost squared becoming gradually longer, about twice as long as wide; 17–19 transverse rows from interbrachials (excluded) to preanals. Five scales in precloacal region, the central one being the smallest. Males usually show four small inconspicuous precloacal pores and two small femoral pores. Pores absent in females.

FIGURE 3. Individuals of (A) Anotosaura collaris adult, and (B) juvenile, and its congener Anotosaura vanzolinia (C), in life.

Tail scales are shorter and more imbricate antero-posteriorly than midbody dorsals, otherwise identical to them, disposed in regular transverse circles, lanceolate; those from ventral part of tail base wider, becoming gradually identical in size around tail. Tail regenerated with rectangular, juxtaposed and smooth scales. Forelimbs extremely robust with large, smooth and imbricate scales, larger and flat dorsally; those from ventral part of brachium smaller. Forearm as long as thick. Anterior and ventral parts of hind limbs with irregularly large, smooth and imbricate scales, largest scales ventrally. Posterior part of hindlimbs with granular and juxtaposed scales. Carpal and tarsal scales large, imbricate; supradigital lamellae smooth, imbricate. Palmar and plantar surfaces with smooth, small granules; infradigital lamellae mostly divided, 6–8 on Finger IV and 12–16 on Toe IV. Fingers and toes clawed, respectively with the following relative sizes: 1 < 2 = 5 < 4 < 3 and, 1< 5 < 2 < 3 < 4. Dorsal surfaces of body and tail light brown with an irregularly distributed dark brown reticulate pattern. Lateral parts of body and tail dark brown with an irregular light brown to cream reticulum. An irregular light line about one scale width extends dorsolaterally from posterior part of head to the middle of tail and becomes inconspicuous toward the tip. Ventral parts of body and tail cream but mottled by a scattered irregular dark brown pattern; tail become gradually darker distally. Dorsal parts of head with an irregular reticulate dark brown pattern.

352 · Zootaxa 3731 (3) © 2013 Magnolia Press RODRIGUES ET AL. Lateral parts of head predominantly dark brown with scattered cream spots concentrated in the suture of labials; ventral parts of head lighter, cream, strongly mottled by an irregular dark brown pigmentation, especially in the external parts. Limbs dark brown, with scattered light brown marks, cream ventrally. Palmar and plantar surfaces light brown. Maximum SVL: 44 mm (females); 41 mm (males). Tail length about 1.5 times SVL in adult males and females. Hemipenial description. The left hemipenis of MZUSP 103845 (Fig. 4A) is evidently bilobed, relatively small, extending along approximately five rows of subcaudals (approx. 3.5 mm). The hemipenial body is roughly Y-shape. The lobes are detached from the body by a marked basal constriction and are completely nude, presenting a round tip (one of the lobes is partially everted getting and artifactual sharp tip).

FIGURE 4. Sulcate, lateral and asulcate faces of the left hemipenis of (A) Anotosaura collaris (MZUSP 103845) and (B) A. vanzolinia (MZUSP 95328). Scale bars = 1mm.

REDISCOVERY OF ANOTOSAURA COLLARIS Zootaxa 3731 (3) © 2013 Magnolia Press · 353 FIGURE 5. Skeleton of Anotosaura collaris (MZUSP 103837), snout-vent length 42.8 mm. (A) Dorsal, (B) lateral, and (C) ventral views of the skull. (D) Ventral view of the pectoral girdle and forelimbs. (E) Ventral view of the pelvic girdle and hindlimbs. Abbreviations: bo, basioccipital; cl, clavicle; cm, columella; cn, coronoid; d, dentary; ept, epipterygoid; exc, exoccipital; f, frontal; fe, femur; fi, fibula; icl, interclavicle; is, ischium; j, jugal; m, maxilla; n, nasal; obp, orbitosphenoid; or, occipital recess; p, parietal; pa.pr, alar process of the prootic; pb, pubis; pd.p, descending process of the parietal; pf, prefrontal pl, palatine; pm, premaxilla; pof, postorbitofrontal; pt, pterygoid; q, quadrate; ra, radius; s.v, sacral vertebra; scc, scapulocoracoid; so, supraoccipital; spc, suprascapula; sq, squamosal; sra, surangular; st, sternum; st.r, sternal rib; ti, tibia; ul, ulna; v, vomer.

The sulcus spermaticus is a relatively broad channel originating at central region of the base of the organ and emarginated by distinctive thickened lips in the basal region. From the base the sulcus spermaticus proceeds in a straight line towards the lobular crotch. At the lobular crotch the sulcus is divided in two shallow branches that run through the internal faces of the lobes and end slightly before the lobular tips. The hemipenial body is ornamented by 14 transversal nude flounces that lack any vestige of mineralized spines or spinules, even after their immersion on Alizarin Red solution for 24 hours. The five more basal flounces are continuous and occupy all the central region of the asulcate face, whereas the remaining and more apical flounces are shorter and interrupted by a nude region in the sagittal region of asulcate face. In both lateral faces of the organ only the four more basal flounces are continuous, whereas the ten remaining are separated by a nude area that gets broader in direction of the distal region. There is a puncture in the asulcate face of the organ, in the region between the first and fifth transversal flounces, probably made accidentally during the hemipenial eversion or filling. The hemipenis of Anotosaura collaris is similar with the organ of A. vanzolinia described and illustrated by

354 · Zootaxa 3731 (3) © 2013 Magnolia Press RODRIGUES ET AL. Nunes (2011). However, the hemipenis of A. collaris lacks the evident calcified and hook-shaped spines present in the 10 more apical flounces of the sulcate face and the vestiges of calcified structures present in some flounces of the asulcate face of the hemipenis of A. vanzolinia (Fig. 4B). Osteology. (Fig. 5) The premaxilla is short, approximately as long as wide (Fig. 5A). Its alveolar plate bears 10 unicuspid pleurodont teeth and contacts laterally the anteromedial process of the maxilla (Fig. 5B). The short nasal process extends dorsally and overlaps the anterior end of the nasals. The palatal shelf of the maxilla bears 10 teeth and is overlapped posteriorly by the maxillary process of the palatine. Its orbital process supports the maxillary process of the jugal. The facial process extends dorsally to meet the nasal, frontal, and prefrontal (Fig. 5A). The vomer (Fig. 5C) has a well-developed dorsal crest that holds, together with the premaxilla and maxilla, the Jacobson's organ. It extends in an anterior process that rests on a pit in the palatal plate of the premaxilla. Posteriorly, it contacts the ventral surface of the palatine. The dome-shaped septomaxilla, located between the vomer and the nasal process of the premaxilla, covers the Jacobson's organ dorsally and laterally. A lateral maxillary process contacts the dorsal surface of the palatal shelf of the maxilla. The nasal rests on the dorsal surface of the septomaxilla. The palatine has a wider anterior portion composed of a medial vomerian process meeting the vomer and a lateral maxillary process contacting the palatal shelf of the maxilla; a pronounced concavity between these processes forms the choanal canal (Fig. 5C). The palatine ends in a pterygoid process. The large prefrontal (Fig. 5A) forms the anterior margin of the orbit. It is overlapped by the facial process of the maxilla anteriorly, and extends posteriorly in a well-developed posterodorsal process, delimiting part of the dorsal margin of the orbit. A large lacrimal foramen and a short lacrimal flange are present. The lacrimal was not observed. The anterior margin of the frontal overlaps the posterior end of the nasals. A distinct anterolateral process is observed laterally to the nasal. The posterior margin of the frontal bears two fronto-parietal tabs that overlap corresponding surfaces of the parietal (Fig. 5A). A posterolateral process contacts the postorbitofrontal. A stout postorbitofrontal is present (Figs. 5A, B), probably corresponding to the fused postorbital and postfrontal. This element forms part of the posterodorsal margin of the orbit and, together with the squamosal, forms the lower margin of the upper temporal fenestra (Fig. 5A). The posterior end of the squamosal fits into a notch in the tympanic crest of the quadrate. The parietal meets the postorbitofrontal anterolaterally. Posteriorly, the parietal forms the anterior margin of the posttemporal fenestra, the posterior margin of which is delimited by the supraoccipital. The postparietal process, which extends posteriorly along the transverse ridge of the supraoccipital, meets the squamosal and supratemporal. A long and slender descending process of the parietal approaches the epipterygoid (Fig. 5B). The jugal forms the lower and part of the posterior margins of the orbit, contacting the maxilla and postorbitofrontal (Fig. 5B). A large maxillary foramen is seen in its medial portion. The jugal bears a medially directed ectopterygoid process that contacts the ectopterygoid. The large Y-shaped pterygoid (Fig. 5C) contacts, at its anterior end, the palatine and the stout ectopterygoid. Medially it contacts the basipterygoid process from the parabasisphenoid and posteriorly, the quadrate. The rod-like epipterygoid has an expanded dorsal end, and its ventral end rests on the dorsal surface of the pterygoid (Fig. 5B). The quadrate (Fig. 5B) has a cephalic condyle that contacts the supratemporal, squamosal, and the paroccipital process, a lateral conch with a slight tympanic crest, and a mandibular condyle that articulates with the mandible. The suborbital fenestra is delimited by the maxilla, vomer, palatine, pterygoid, and ectopterygoid. The upper temporal fenestra is delimited by the postorbital, postfrontal, parietal, and squamosal. The chondrocranium is composed of the supraoccipital, basioccipital, exoccipital, opisthotic, otooccipital, and parabasisphenoid. The supraoccipital closes the roof of the braincase and forms the dorsal margin of the foramen magnum. Laterally, it contacts the prootic-otoccipital complex. The posterior and horizontal semicircular canals are observed within this bone. The prootic-otooccipital complex is located laterally, forming the otic capsule. The prootic is the anterior element and the otooccipital is the posterior one, and both delimit the fenestra ovalis, to which the columella is inserted (Fig. 5C). The anterior semicircular canal is seen in the alar process (Fig. 5B) of the anterior margin of the prootic. The incisura prootica, below the alar process, forms a C-shaped exit for the trigeminal nerve. The facial foramen is located posteriorly to the incisura prootica. The crista prootica extends ventrally from this latter foramen to the parasphenoid. The vestibular and lagenar cavities of the prootic are distinct in the inner surface of the prootic.

REDISCOVERY OF ANOTOSAURA COLLARIS Zootaxa 3731 (3) © 2013 Magnolia Press · 355 The otooccipital is composed of the fused opisthotic and exoccipital; the occipital recess and vagus foramen are delimited anteriorly by the opisthotic and posteriorly by the exoccipital. The anterodorsal margin of the opisthotic forms the well-developed paroccipital process. The exoccipital contributes to the occipital condyle. Three foramina for the hypoglossal nerve are present. The ventral basioccipital, which forms most of the occipital condyle, is fused to the parabasisphenoid, located anteriorly (Fig. 5C), and contacts the prootic and otooccipital laterally. The tetraradiate orbitosphenoid is connected to the interorbital cartilages. The mandible (Figs. 5B, C) is formed by the dentary, splenial, angular, surangular, coronoid, and prearticular- articular complex. The dentary bears 15 teeth. Its labial surface (Fig. 5B) contacts the splenial and angular ventrally, the surangular posteriorly, and overlaps the labial process of the coronoid. The lingual surface of the dentary is overlapped by the splenial and by the anteromedial process of the coronoid. The coronoid bears a high dorsal process (Fig. 5B), and a posteromedial process that overlaps the prearticular, forming the anterior margin of the adductor fossa. The surangular is located posteriorly on the labial surface of the mandible, forming the external margin of the adductor fossa. The prearticular-articular complex forms most of the posterior end of the mandible and the floor and posterior margin of the adductor fossa. The hyoid apparatus (Fig. 5C) is composed of a short basihyal, a glossohyal extending anteriorly, and three pairs of visceral arches: the hyoid cornu and the associated epihyal originating from its mid portion, and the first and second ceratobranchials. The specimen analyzed has 28 presacral vertebrae. Nine intercentra are present, associated with the vertebral centrum of the most anterior vertebrae. Ribs are present from the 4th vertebra on, and the last presacral vertebra does not bear ribs. The scapular girdle (Fig. 5D) is composed of a stout clavicle that bears a large foramen on its medial end, a rod-like interclavicle, the scapulocoracoid with four foramina, and the suprascapula. A sternum with a large central foramen, and a rod-like xiphisternum are present. The sternum is associated with two pairs of sternal ribs and the xiphisternum is associated with a single pair of xiphisternal ribs. The forelimb (Fig. 5D) is composed of a humerus with well-developed proximal and distal heads, a stout ulna and a slender radius, and a series of carpals and phalanges. The carpal region is composed of ulnare, radiale, pisiform, palmar, central, and four distal carpals (from II to V). The phalangeal formula of the forelimb is 2:3:4:4:3. The pelvic girdle (Fig. 5E) is formed by the pubis, ischium, and ilium. The pubis has a large obturator foramen and a well-developed pectinal process. Epipubis, epiischium and hypoischium are present. The hindlimb (Fig. 5E) is formed by the femur, tibia, and fibula. A large astragalus-calcaneum and smaller distal tarsals III and IV are present in the tarsal region. Two ventral sesamoids are also present. The phalangeal formula of the hindlimb is 2:3:4:5:3. Morphological variation and morphometry. Table 1 presents comparative data on external morphology of Anotosaura collaris, A. vanzolinia, Colobosauroides cearensis, C. carvalhoi Soares & Caramaschi, 1998, and Dryadosaura nordestina. Table 2 shows morphometric variation in A. collaris and comparisons among males and females. Sexes were not significantly different in SVL (P = 0.394) but sexual dimorphism was detected in relative size, with females presenting larger LBL (P < 0.01) and males presenting larger HW (P < 0.01) (Table 2). Sexual dimorphism did not interacted with species differences (P > 0.05 for all characters) thus sexes were pooled for intrageneric comparisons. Although similar in body size, A. vanzolinia had shorter limbs, with significantly smaller FEM, HUM and FAL (P < 0.01), a narrower head with smaller HW (P < 0.01) and it was slightly but significantly more elongated than A. collaris, presenting proportionally larger LBL (ANCOVA with SVL as covariate, P < 0.01). A discriminant analysis including only males of the two species of Anotosaura plus C. cearensis and D. nordestina shows that the first discriminant function explains 93.5% of variation among species, with high and positive loadings for limb length (FAL, HUM, TIB, and FTL) (Table 3). In the morphological space defined by this axis, both Anotosaura species are well differentiated from Colobosauroides cearensis and Dryadosaura nordestina (Fig. 6), presenting relatively shorter forelimbs and hindlimbs. The four species are correctly classified by the discriminant function with 100% of correct classification for Anotosaura collaris and C. cearensis, 91.7% for A. vanzolinia (one individual was classified as A. collaris) and 87.5% for D. nordestina (one individual was classified as C. cearensis). The second discriminant function explains only 4.9% of variation and present positive loadings for all variables (Table 3).

356 · Zootaxa 3731 (3) © 2013 Magnolia Press RODRIGUES ET AL. TABLE 1. Comparative table of external morphological characters in species of Anotosaura, Dryadosaura and Colobosauroides. Meristic traits are shown as ranges. SVL, TAL, and TAL/SVL are shown as mean ± standard deviation. A. collaris A. vanzolinia C. cearensis C. carvalhoi D. nordestina Dorsals 26–29 28–31 23–24 24–25 23–25 Scales around midbody 23–25 24–26 29–31 33–34 16–33 Ventrals 18–20 19–23 14–16 15–16 15–17 Gulars 5 5 4–5 6 5 Pre-anal pores (males / females) 4/0 4/0 4/0 4/0 4/2–4 Femoral pores (males) 2–4 3–6 3–4 4 5–8 Lamellas under fourth finger 6–8 5–8 8–10 - 6–9 Lamellas under fourth toe 12–16 12–16 14–17 15–17 13–16 Supralabials 6 6 6 6 6 Infralabials 5–6 6 6–7 7 5 Supraoculars 3 3 3 3 3 Superciliars 2–4 2–3 4–5 3 4 Suboculars 2–4 1–3 4 4 2 SVL 39.4 ± 3.7 40.3 ± 5.5 38.0 ± 4.5 35.9 ± 4.0 48.8 ± 4.7 TAL (TAL/SVL) 58.7 ± 3.1 65.3 ± 9.1 58.8 ± 5.5 51.5 ± 5.5 78.2 ± 9.1 (1.50 ± 0.03) (1.53 ± 0.04) (1.54 ± 0.10) (1.48 ± 0.09) (1.53 ± 0.12)

TABLE 2. Morphometric measurements of females and males of Anotosaura collaris (average ± standard deviation) and results of ANOVA/Kruskal-Wallis test for sexual dimorphism in each character. SVL=snout-vent length, LBM=length between limbs, BW=body width, HH=head height, HW=head width, HL=head length, FEM=femur length, TIB=tibia length, FTL=foot length, HUM=humeral length, FAL=forearm length, FAW=forearm width.

Average ± Standard Deviation ANOVA Kruskal-Wallis Females (N = 6) Males (N = 6) FP P SVL 41.3 ± 2.4 39.4 ± 0.5 - - 0.394 LBM 26.5 ± 1.4 24.4 ± 0.4 - - 0.002 BW 3.5 ± 0.2 3.4 ± 0.1 0.08 0.778 - HH 2.9 ± 0.2 3.1 ± 0.1 3.26 0.101 - HW 4.4 ± 0.1 4.7 ± 0.1 15.06 0.003 - HL 6.4 ± 0.3 6.6 ± 0.2 2.31 0.160 - FEM 3.2 ± 0.2 3.3 ± 0.1 1.53 0.244 - TIB 3.0 ± 0.2 3.1 ± 0.1 1.28 0.285 - FTL 4.3 ± 0.2 4.5 ± 0.2 1.97 0.191 - HUM 1.9 ± 0.1 1.9 ± 0.04 1.51 0.248 - FAL 3.9 ± 0.1 3.9 ± 0.1 0.21 0.654 - FAW 1.1 ± 0.1 1.1 ± 0.02 - - 0.485

Karyotype. The karyotype of Anotosaura collaris is composed by 44 biarmed chromosomes, 20 macrochromosomes and 24 microchromosomes (Fig. 7). Pairs 1, 4, 5, 6, 8 and 10 are metacentric and pairs 2, 3, 7, and 9 are submetacentric. Molecular Phylogeny. Bayesian and Maximum Likelihood analysis recovered the same topology; Ecpleopus gaudichaudii Duméril & Bibron, 1839 as an early-diverging lineage within Ecpleopodini, sister to two distinct clades. One clade with (Colobosauroides cearensis + (Anotosaura + Dryadosaura), and the other grouping Marinussaurus, Leposoma and Arthrosaura, with unresolved relationships. The analysis recovered Anotosaura as monophyletic (PP=1, ML bootstrap=98) (Fig. 8).

REDISCOVERY OF ANOTOSAURA COLLARIS Zootaxa 3731 (3) © 2013 Magnolia Press · 357 FIGURE 6. Results of a discriminant analysis on morphometric measurements of male individuals of Anotosaura collaris (blue circles), A. vanzolinia (green circles), Colobosauroides cearensis (orange diamonds) and Dryadosaura nordestina (red squares). Group centroids are represented by a black dot. In parenthesis is the amount of original variation explained by each axis.

FIGURE 7. Karyotype of Anotosaura collaris after conventional staining, showing 2n=44 (20M + 22m).

358 · Zootaxa 3731 (3) © 2013 Magnolia Press RODRIGUES ET AL. TABLE 3. Results of a discriminant function analysis on morphometric data of males of Anotosaura collaris, A. vanzolinia, Colobosauroides cearensis and Dryadosaura. nordestina. SVL=snout-vent length, LBL=length between limbs, BW=body width, HH=head height, HW=head width, HL=head length, FEM=femur length, TIB=tibia length, FTL=foot length, HUM=humeral length, FAL=forearm length, FAW=forearm width. Variables DF 1 DF 2 FAL 0.475 0.463 HUM 0.329 0.252 TIB 0.319 0.403 FTL 0.314 0.378 FEM 0.296 0.306 FAW 0.264 0.211 HW 0.260 0.372 HL 0.252 0.202 HH 0.187 0.331 BW 0.154 0.190 SVL 0.074 0.405 LBL -0.012 0.422 Eingenvalue 77.58 4.03 % of Variance 93.5 4.9 Cumulative % 93.5 98.4

Uncorrected p-distances varied from 2.2 to 19.3% for 16S, 3.7 to 23.8% for 12S, 8.8 to 28.9% for ND4 and 4.0 to 27.8% for C-mos among Gymnophthalmidae sampled. Distances between Anotosaura collaris and A. vanzolinia varied from 2.2 to 2.8% for 16S, 3.7 to 4% for 12S and 8.8 to 9.4% for ND4 and while distances among A. collaris specimens were nearly zero (Table 4). Distribution and natural history. Our sampling efforts in the present and as well as previous field trips covered a reasonably wide range of habitats and areas in the Caatinga. However, Anotosaura collaris was only found in three localities: Senhor do Bonfim, Missão do Sahy, and Campo Formoso (Fig. 9). Maximum straight-line distance between them is about 15km. All sites are situated in isolated relictual mountains, emerging from the general Caatinga area, in the northern end of Serra de Jacobina, a small branch of Chapada Diamantina. Other lizards obtained at the same sites were the gymnophthalmids Acratosaura mentalis and Psilophthalmus sp., the phyllodactylid Gymnodactylus geckoides, the gekkonid Hemidactylus brasilianus, the teiid Ameivula ocellifera, and the tropidurids Tropidurus hispidus and T. semitaeniatus. This region has a basement formed by Paleoproterozoic and Neoarchean (2800–1600 MYbp) sedimentary metamorphic rocks (Nápravník 2011) and rises up to 1200 m a.s.l. The soils occupied by A. collaris are clayish, different from those occupied by all other species analyzed here, which tend to be more sandy (Table 5). These mountains are a barrier for the wet winds coming from the coast, which is congruent with the low precipitation seasonality at these sites (Table 5), and the presence of cloud forests at higher elevations. At the top, those mountains are often covered with campos rupestres, bearing extensive rock outcrops, whereas the lower flatlands are covered with typical thorny-shrubby Caatinga vegetation. In Senhor do Bonfim, specimens of Anotosaura collaris were found at Alto da Rainha, an isolated mountain at about 715 m a.s.l. elevation. In Missão do Sahy, an old indian Mission, specimens were found on the top of a mountain at 895 m a.s.l. and in Campo Formoso, at Morro do Cruzeiro (780 m a.s.l.). Two A. collaris’ shed skins were also found in Coqueiral (10°33'9.06"S, 40° 8'42.26"W), another isolated mountain at about 593 m a.s.l.. All specimens were collected by active search, using rakes. Anotosaura collaris shows semi-fossorial behavior. Individuals were found only in the sub-superficial stratum, under leaf litter or small rocks. The maximum lizard density found was a group of two adults and two juveniles collected under a bush covering an area of approximately 32 m2.

REDISCOVERY OF ANOTOSAURA COLLARIS Zootaxa 3731 (3) © 2013 Magnolia Press · 359 360 · Zootaxa 3731 (3) © 2013 Magnolia Press RODRIGUES ET AL. Averaged thermal limits for Anotosaura collaris were: critical thermal maximum (CTmax) = 38.78 oC (standard deviation = 0.6, N=7) and critical thermal minimum (CTmin) = 14.81 oC (sd=2.26, N=7) (Table 5). During the period sampled, environmental temperatures over and under the leaf litter never reached A. collaris’ CTmin or CTmax. However, on rock surfaces or under rocks, temperatures exceeded CTmax (Fig. 10), thus this microhabitat is probably avoided during the day; although we found some individuals under rocks but always in shaded areas. TABLE 5. Comparison of ecogeographical traits among five related Ecpleopodini species from northeastern Brazil. N represents the number of available geographic records at which climatic, topographic and soil traits were extracted from climatic and soil databases. Climatic means are calculated from measurements made during at least 10 years of climatic data. A. collaris A.vanzolinia C. cearensis C. carvalhoi D. nordestina N=5 N=15 N=9 N=1 N=17 Soil texture class 3.00±00 11.34±1.40 11.00±0.50 13.00 10.50±1.17 Altitude 565.28±78.69 490.69±185.83 471.69±288.56 546.30 162.86±179.95 Annual mean temperature 23.12±0.55 23.08±1.24 23.57±1.92 24.34 24.26±1.38 Diurnal temperature range 10.57±0.06 9.30±0.50 8.81±1.49 14.18 7.78±0.85 Maximum temperature 30.64±0.63 30.07±1.20 29.44±2.26 33.81 29.88±1.33 of the warmest month Minimum temperature 16.02±0.65 16.67±1.26 17.97±1.92 13.9 18.66±1.31 of the coldest month Mean annual precipitation 763.85±46.67 624.68±111.97 1397.45±118.3 1083.76 1496.85±377.43 Mean precipitation seasonality 36.50±1.38 65.79±19.99 87.15±6.48 86. 015 58.61±13.10

FIGURE 8. Phylogenetic relationships recovered through a Bayesian (BA) and Maximum Likelihood (ML) analysis of Anotosaura collaris based on mitochondrial (12S, 16S and ND4) and nuclear genes (C-mos and 18S). The value for posterior probabilities (BA), and bootstrap (ML) are show on branches, respectively.

REDISCOVERY OF ANOTOSAURA COLLARIS Zootaxa 3731 (3) © 2013 Magnolia Press · 361 FIGURE 9. Currently known distribution of Anotosaura collaris (left) and of Anotosaura, Colobosauroides and Dryadosaura (right). On the right, colors represent different biomes.

FIGURE 10. Comparison of environmental temperatures at microhabitats used by Anotosaura collaris with its critical thermal limits. CTmax (red) and CTmin (blue) are species means. Dots around the boxplots represent outliers. Whiskers end at the 5th (below) and and the 95th (above) percentiles. Horizontal lines within the box plot represent the 25th, 50th and 75 quartiles. Temperatures measured in December 2012.

362 · Zootaxa 3731 (3) © 2013 Magnolia Press RODRIGUES ET AL. Discussion

Over one hundred years have passed since the collection of the holotype of Anotosaura collaris by Ernest Garbe, and until now no other individual had ever been found. This fact reinforces the idea that despite the increasing effort and methodological improvements on herpetofaunal surveys at the Caatinga domain, they are still far from being comprehensive (Rodrigues 2003). Nonetheless, our directional efforts on the surroundings of the type locality allowed us to find 16 specimens, two shed skins, and a broken tail, probably from an additional lizard lost during the searching process. Our first challenge was to locate the type locality, described as “Villa Nova da Rainha” (currently municipality of Senhor do Bonfim) by Ernest Garbe (Garbe 1908; Ihering & Ihering 1911; Amaral 1933). The letters Garbe exchanged with Hermann von Ihering, the director of the Museum Paulista at that time, indicate that his first field site was a ranch situated about 6 km from Senhor do Bonfim, where the dominant vegetation was typical dry Caatinga. He later moved to another site, about 10 km from the first one, dominated by forest (Garbe 1908). Unfortunately, a better association of Garbe’s collecting sites to any present location is not possible based on the geographical references available. His first site might have been in flatland areas or even localities westwards from the city of Senhor do Bonfim, encompassing part of the Jacobina range, with mountains similar to those where Anotosaura collaris occurs. Nonetheless, it is not unlikely that E. Garbe visited Alto da Rainha, as the mountain sticks out from the general landscape, calling the attention of any one who passes by (Fig. 11). Besides the uncertainty regarding the exact location of the type locality, there is also some controversy regarding some scale characters observed in the type specimen. Amaral (1933) reports three supraoculars, two suboculars, 25 dorsal and 18 ventral rows. However, Dixon (1974), who also examined the holotype, reports three supraoculars, one subocular, 27 dorsal and 19 ventral rows. The latter author included the interbrachial row in his count of the ventral scales and states that Amaral must have found only two supraoculars instead of the three reported. However, the holotype in Amaral’s description clearly shows three supraoculars. To clear out these misunderstandings, we reexamined the poorly preserved holotype, and confirm the presence of three supraoculars, two supraciliars, two suboculars, 26 dorsal rows and 18 ventrals. The discrepancies regarding the ventrals could be due to the inclusion, by Dixon, of the pectoral shield in his counts. Differences in the number of dorsal rows are possibly due to an asymmetry in the arrangement of the second row of these scales: the first transverse dorsal row following the parietal/interparietal edge is regularly organized but the second one has an enlarged paravertebral scale at its left side, which corresponds to two scales on the right, thus dorsal counts are distinct if taken on the left or the right sides. The new specimens obtained bring additional information regarding intraspecific variability, showing that the number of dorsals varies from 26 to 29 scales. Scale counts, along with morphometric data, showed that morphology is highly conserved among Anotosaura, Colobosauroides, and Dryadosaura (Fig. 6; Table1). This is consistent with the recovery of a well-supported clade including these species by both molecular and morphological data (Pellegrino et al. 2001; Castoe et al. 2004; Rodrigues et al. 2005; Peloso et al. 2011). However, although the monophyly of Ecpleopodini has been recovered in previous molecular phylogenies (Pellegrino et al. 2001; Castoe et al. 2004; Rodrigues et al. 2005; Peloso et al. 2011), the relationships among the three genera are not resolved. Pellegrino et al. (2001) recovered Anotosaura as sister to Dryadosaura, although with low support, and Colobosauroides as sister to both in a concatenated maximum parsimony (MP) analysis. In their maximum likelihood analysis (Pellegrino et al. 2001), Anotosaura was recovered, also with low support, as sister to Colobosauroides, and Dryadosaura as sister to both taxa. Using a Bayesian analysis, Castoe et al. (2004) recovered Dryadosaura sister to Colobosauroides + Anotosaura and, once more, the monophyly of the clade formed by the three species was highly supported but the relationship between the two sister taxa was not. Rodrigues et al. (2005) and Peloso et al. (2011) recovered Anotosaura + Dryadosaura sister to Colobosauroides, both in morphological and in combined morphological and molecular MP trees. Our Bayesian analysis recovered Colobosauroides as sister to Anotosaura + Dryadosaura. The monophyly of Anotosaura, which had never been tested before, was also corroborated by our results. This was expected given the high morphological similarity between A. collaris and A. vanzolinia, especially in conspicuous features such as the absence of ear opening, only shared by Anotosaura species among Ecpleopodini. The Gymnophthalmidae are known for their extensive karyotypic variability in diploid number, varying from 2n = 32 in Bachia dorbignyi to 2n = 62-64 in Nothobachia ablephara Rodrigues, 1984 (Pellegrino et al. 1999b, 2001). Chromosome size and shape varies, with some species showing marked differences between macro- and

REDISCOVERY OF ANOTOSAURA COLLARIS Zootaxa 3731 (3) © 2013 Magnolia Press · 363 FIGURE 11. Habitat of Anotosaura collaris: (A) general view of Morro do Cruzeiro, Campo Formoso municipality; (B) landscape at Senhor do Bonfim, showing the city at the right and Alto da Rainha hill (arrow) at the center; (C) mountain range at Missão do Sahy and (D) detail of the vegetation at the top of Alto da Rainha hill, Senhor do Bonfim, Bahia. microchromosomes, while other show gradual variation in size. There is also variation in the presence of supernumerary chromosomes, as well as in ploidy level, with diploid and triploid species in the family. Gymnophthamidae are also diverse in terms of systems of chromosomal sex determination (Cole et al. 1990, 1993; Yonenaga-Yassuda & Rodrigues 1999; Yonenaga-Yassuda et al. 1995, 1996, 2005; Pellegrino et al. 1999a, b, 2003; Laguna et al. 2010a, b). The karyotype of Anotosaura collaris is composed by 2n=44 (20M +24m), similar to other Gymnophthalmini, Ecpleopodini and Cercosaurini (Sherbrooke & Cole 1972; Cole et al. 1990, 1993; Yonenaga-Yassuda et al. 1995, 2005; Pellegrino et al. 1999, 2001; Laguna et al. 2010a, b). However, despite their similar macrochromosome morphology, chromosome arm lengths differ among species, suggesting that events of inversion are involved and could have been responsible for their differentiation. Among Ecpleopodini, three other diploid numbers were

364 · Zootaxa 3731 (3) © 2013 Magnolia Press RODRIGUES ET AL. described: 2n=46 (22M + 24m) in Anotosaura vanzolinia; 2n=48 (20M +28m) in Dryadosaura nordestina; and 2n=52 in Leposoma scincoides Spix, 1825 (Pellegrino et al. 1999, 2001; Laguna et al. 2010a, b). The striking chromosomal differences between A. collaris and A. vanzolinia indicates the occurrence of Robertsonian rearrangements. Likewise, the differences in diploid numbers in Ecpleopodini are probably due to fusion/fission mechanisms. Karyotypes of Ecpleopus, Arthrosaura and Marinussaurus are unknown, precluding additional considerations on chromosomal evolution in this tribe. As suggested by morphological evidences, (absence of an external ear opening, elongated body, reduced eye size, and short and robust forelimbs), soil type, and microhabitat use, Anotosaura collaris is fossorial or semifossorial, living in relatively open habitats, in the Caatinga Domain. The coupled evolution of body elongation and limb reduction is a major morphological trend in Gymnophthalmidae (Rodrigues et al. 2013; Wiens et al. 2006; Grizante et al. 2012). The intersexual pattern of variation observed in A. collaris is recurrent in the family, with female presenting larger trunks, possibly related to the storage of eggs, and males with larger heads, that might confer advantage in male-male combats for mates (Pianka & Vitt 2003; Balestrin et al. 2010). Our records indicate that Anotosaura collaris is predominantly associated with habitats in the highest elevations of its known distributional range, surrounded by lowlands with typical Caatinga vegetation. At the mountaintops temperatures are cooler than those prevalent at lower regions Caatingas due to the adiabatic gradient. Anotosaura collaris’ distribution might be wider than reported here, given the presence of several mountains in this region harboring similar habitats than those found in Senhor do Bonfim, Missão do Sahy, and Campo Formoso. Curiously, despite the apparent current isolation of the populations in mountains by unsuitable microclimates in the intervening Caatinga flatlands, genetic divergence among A. collaris’ localities are low, suggesting recent connectivity among populations. One possible explanation is that current geographic isolation among mountaintop localities has not yet been reflected on genetic distances, because milder climate during the Last Glacial Maximum (21 kyBP) might have allowed A. collaris to occupy lower as well as high altitude areas. Currently the lowlands are occupied only by A. vanzolinia. Another potential explanation is that most suitable microhabitats for this species at the lowlands have been recently devastated by human’s action. We uncovered many facts that highlight the potential vulnerability of Anotosaura collaris to extinction in the near future. Anotosaura collaris shows a restricted distribution range, relative low density, and uses microhabitats that show patchy distribution restricted to mountain tops. This is even more concerning given that we observed intense anthropogenic habitat alteration at, and around, these mountaintops during the period we were collecting in the area (i.e. expansion of human settlements, mining, and frequent fires). Based on these facts, we strongly recommend further extinction risk assessments for this species, urging for more surveys covering broader areas. Further understanding of its ecological and physiological requirements and its ecogeographical traits will certainly aid the delineation of conservation policies for this species. A few apparently still preserved patches at Serra da Jacobina, might constitute the last big tract of habitat for A. collaris.

Acknowledgements

The authors were funded by Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP), Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq) and PROEX Program of Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES). Roberta Damasceno acknowledges funding from CAPES-Fulbright (BEX2740/06-0). We thank Hussam Zaher (MZUSP), Carolina Castro-Mello (MZUSP), and Paulo Passos (MNRJ) for access to specimens, Federico Arias, Tais Machado and Rangel Batista de Carvalho for help in the field, and Enio Mattos, Phillip Lenktaitis, Sabrina Baroni and Maysa Miceno for logistic support.

Literature cited

Amaral, A. (1933) Estudos Sobre Lacertilios Neotropicos I. Novos generos e especies de largartos do Brasil. Memorias do Instituto Butantan, 7 (1932), 51–74. Amaral, A. (1937) Check-list of the Lacertilia from Brasil. Comptes Rendus XII Congrés International de Zoologie (Lisbonne 1935), 1733–17343. Amaral, A. (1938) Estudos sobre lacertílios neotropicos. 4. Lista remissiva dos lacertílios do Brasil. Memórias do Instituto Butantan, 11 (1937), 167–204.

REDISCOVERY OF ANOTOSAURA COLLARIS Zootaxa 3731 (3) © 2013 Magnolia Press · 365 Arévalo, E.S., Davis, S.K. & Sites, J.W. Jr. (1994) Mitochondrial DNA sequence divergence and phylogenetic relationships among eight chromosome races of the Sceloporusgrammicus complex (Phrynosomatidae) in central Mexico. Systematic Biology, 43, 387–418. Balestrin, R.L., Cappellari, L.H. & Outeiral, A.B. (2010) Biologia reprodutiva de Cercosaura schreibersii (Squamata, Gymnophthalmidae) e Cnemidophorus lacertoides (Squamata, Teiidae) no Escudo Sul-Riograndense, Brasil. Biota Neotropica, 10, 131–139. http://dx.doi.org/10.1590/s1676-06032010000100013 Bell, C.J., Evans, S.E. & Maisano, J.A. (2003) The skull of the gymnophthalmid lizard Neusticurus ecpleopus (Reptilia: Squamata). Zoological Journal of the Linnean Society, 139, 283–304. http://dx.doi.org/10.1046/j.1096-3642.2003.00077.x Bogart, J.P. (1973) Method for obtaining chromosomes. Caldasia, 11, 29–40. Borges-Nojosa, D.M. & Caramaschi, U. (2003) Composição e Análise Comparativa da Diversidade e das Afinidades Biogeográficas dos Lagartos e Anfisbenídeos (Squamata) dos Brejos Nordestinos. In: Leal, I., Silva, L.M.C. & Tabarelli, M. (Eds.), Ecologia e Conservação da Caatinga. UFPE, Recife, pp. 463–512. Brooks, G.R. & Sassman, J.F. (1965) Critical thermal maxima of larval and adult Eurycea bislineata. Copeia, 1965, 251–251. Camacho, A.C. & Rodrigues, M.T. (2006) Dryadosaura nordestina (Briba Cabeçuda). Geographic Distribution. Herpetological Review, 38, 218. Castoe, T.A., Doan, T.M. & Parkinson, C.L. (2004) Data partitions and complex models in Bayesian analysis: The phylogeny of gymnophthalmid lizards. Systematic Biology, 53, 448–469. Cole, C.J; Dessauer, H.C.; Townsend, C.R. & Arnold, M.G. (1990) Unisexual lizards of the genus Gymnophthalmus (Reptilia, Teiidae) in the neotropics: Genetics, origin, and systematics. American Museum Novitates, 2994, 1–29. Cole, C.J., Dessauer, H.C. & Markezich, A.L. (1993) Missing link found: the second ancestor of Gymnophthalmus underwoodi (Squamata: Teiidae), a South American unisexual lizard of hybrid origin. American Museum Novitates, 3055, 1–13. Cowles, R.B. & Bogert, C.M. (1944) A preliminary study of the thermal requirements of desert reptiles. Bulletin of the American Museum of Natural History, 83, 261–296. http://dx.doi.org/10.1086/394795 Cunha, O.R., Lima-Verde, J.S. & Lima, A.C.M. (1991) Novo gênero e espécie de lagarto do Estado do Ceará (Lacertilia: Teiidae). Boletim do Museu Paraense Emilio Goeldi Serie Zoologia, 7, 163–176. Delfim, F.R. & Freire, E.M.X. (2007) The gymnophtalmid lizards (Squamata: Gymnophthalmidae) of the Cariri region of Paraiba and Seridó of Rio Grande do Norte, northeastern Brazil: considerations about geographic distribution and ecology. Oecologia Brasiliensis, 11, 365–382. http://dx.doi.org/10.4257/oeco.2007.1103.07 Delfino, M.S.C. & Soeiro, M.S. (2012) Dryadosaura nordestina (Briba Cabeçuda). Herpetological Review, 43, 102. Dixon, J. (1973) A systematic review of the teiid lizards, genus Bachia with remarks on Heterodactylus and Anotosaura. Miscellaneous Publications of the Museum of Natural History of the University of Kansas, 57, 1–47. Dixon, J. (1974) Systematic review of the lizard genus Anotosaura (Teiidae). Herpetologica, 30, 13–18. Dowling, H.G. & Savage, J.M. (1960) A guide to the snake hemipenis: a survey of basic structure and systematic characteristics. Zoologica, 45, 17–28. Edgar, R.C. (2004) MUSCLE: multiple sequence alignment with high accuracy and high throughput. Nucleic Acids Research, 32, 1792–1797. Evans, S. (2008) The skull of lizards and tuatara. In: Gans, C., Gaunt, A.S. & Adler, K. (Eds.), Biology of the Reptilia, Vol. 20. Morphology H: The Skull of Lepidosauria. Society for the Study of Amphibians and Reptiles, New York, pp. 1–344. Falcão, A.C.G.P. & Hernández, M.I.M. (2007) Lagartos de restinga no Nordeste brasileiro: análise da fauna de áreas conservadas e reflorestadas. Anais do VIII Congresso de Ecologia do Brasil, 23 a 28 de Setembro de 2007, Caxambu, MG, 1–3. Fetzner, J.W. (1999) Extracting high-quality DNA from shed reptile skins: A simplified method. Biotechniques, 26, 1052– 1054. Freire, E.M.X., Sugliano, G.O.S., Kolodiuk, M.F., Ribeiro, L.B., Maggi, B.S., Rodrigues, L.S., Vieira, W.L.S. & Falcão, A.C.G.P. (2009) Répteis Squamata das Caatingas do seridó do Rio Grande do Norte e do cariri da Paraíba: síntese do conhecimento atual e perspectivas. In: Freire, E.M.X. (Org.), Recursos Naturais das Caatingas: uma visão multidisciplinar. EDUFRN, Natal, pp. 51–84. Freitas, M.A. & Moisés, L.E. (2009) Anotosaura vanzolinia. Geographic Distribution. Herpetological Review, 40 (2), 237. Garbe, E. (1908) letters to H. von Ihering. Fundo Museu Paulista Série - Correspondências 1908. Godinho, R., Crespo, E.G., Ferrand, N. & Harris, D.J. (2005) Phylogeny and evolution of the green lizards, Lacerta spp. (Squamata: Lacertidae) based on mitochondrial and nuclear DNA sequences. Amphibia-Reptilia, 26, 271–285. http://dx.doi.org/10.1163/156853805774408667 Gogliath, M., Ribeiro, L.B. & Freire, E.M.X. (2010) Anotosaura vanzolinia. Geographic Distribution. Herpetological Review, 41, 243. Gonçalves, U., Brito, P.S., Galdino, J.Y. & Torquato, S. (2012) Squamata, Gymnophthalmidae, Anotosaura vanzolinia Dixon, 1974: New records and geographic distribution map. Check List, 8, 632–633. Grizante M.B., Brandt, R. & Kohlsdorf, T. (2012) Evolution of Body Elongation in Gymnophthalmid Lizards: Relationships with Climate. PLoS ONE, 7, 1–7.

366 · Zootaxa 3731 (3) © 2013 Magnolia Press RODRIGUES ET AL. Harvey, M.B. & Embert, D. (2008) Review of Bolivian Dipsas (Serpentes: Colubridae), with comments on other South American species. Herpetological Monographs, 22, 54–105. http://dx.doi.org/10.1655/07-023.1 Hijmans, R.J., Cameron, S.E., Parra, J.L., Jones, P.G. & Jarvis, A. (2005) Very high resolution interpolated climate surfaces for global land areas. International Journal of Climatology, 25, 1965–1978. http://dx.doi.org/10.1002/joc.1276 Hijmans, R.J. & van Etten, J. (2013) Raster: Geographic data analysis and modeling. R package version 2.1-16. Available from: http://CRAN.R-project.org/package=raster (Accessed 2 October 2013) Hoffstetter, R. & Gasc, J.P. (1969) Vertebrae and ribs of modern reptiles. In: Gans, C., Parsons, T.S. & Bellairs, A. d’A. (Eds.), Biology of the Reptilia, Vol. 1. Academic Press, London, pp. 201–310. Ihering, H. & Ihering, R. (1911) O Museu Paulista nos anos de 1906 a 1909. Revista do Museu Paulista, 8, 1–22. http://dx.doi.org/10.1590/s0101-47142008000100006 Jerez, A. & Tarazona, O.A. (2009) Appendicular skeleton in Bachia bicolor (Squamata: Gymnophthalmidae): osteology, limb reduction and postnatal skeletal ontogeny. Acta Zoologica, 90, 42–50. http://dx.doi.org/10.1111/j.1463-6395.2008.00331.x Laguna, M.M., Rodrigues, M.T., Santos, R.M.L., Yonenaga-Yassuda, Y., Ávila-Pires, T.C.S., Hoogmoed, M.S. & Pellegrino, K.C.M. (2010a) Karyotypes of a cryptic diploid form of the unisexual Leposoma percarinatum (Squamata, Gymnophthalmidae) and the bisexual Leposoma ferreirai from the lower Rio Negro, Amazonian Brazil. Journal of Herpetology, 44, 153–157. http://dx.doi.org/10.1670/08-250.1 Laguna, M.M., Garcia, C., Amaro, R.C., Rodrigues, M.T. & Yonenaga-Yassuda, Y. (2010b) Relações evolutivas entre os lagartos da Tribo Ecpleopodini (Gymnophthalmidae, Squamata): uma abordagem cromossômica e molecular. XVIII Encontro de Genética do Nordeste, 2010, Jequié - BA. XVIII ENGENE. Manzani, P.R. & Abe, A.S. (1988) Sobre dois novos métodos de preparo do hemipênis de serpentes. Memórias do Instituto Butantan, 50, 15–20. Miller, M.A., Pfeiffer, W. & Schwartz, T. (2010) Creating the CIPRES Science Gateway for inference of large phylogenetic trees. In: Proceedings of the Gateway Computing Environments Workshop (GCE), 14 November 2010. New Orleans, LA, pp. 1–8. Moura, G.J.B., Santos, E.M., Oliveira, M.A.B. & Cabral, M.C.C. (2011) Herpetofauna no estado de Pernambuco. IBAMA, Brasília, 440 pp. Myers, C.W. & Donnelly, M.A. (2001) Herpetofauna of the Yutaje-Corocoro massif, Venezuela: Second report from The Robert G. Goelet American Museum-terramar expedition to the northwestern tepuis. Bulletin of the American Museum of Natural History, 1–85. http://dx.doi.org/10.1206/0003-0090(2001)261<0001:hotycm>2.0.co;2 Myers, C.W. & Donnelly, M.A. (2008) The summit herpetofauna of Auyantepui, Venezuela: report from the Robert G. Goelet American Museum - TERRAMAR Expedition. Bulletin of the American Museum of Natural History, 308, 1–147. http://dx.doi.org/10.1206/308.1 Nachtergaele, F., van Velthuizen, H., Verelst, L. & Wiberg, D. (2012) Harmonized World Soil Database - Version 1.2. FAO, Rome, Italy and IIASA, Laxenburg, Austria, 42 pp. Available from: http://www.iiasa.ac.at/ (Accessed 2 October 2013) Nápravník, L.A.P. (2011) Características geoquimicas, petrográficas e metalogenéticas dos flogopitititos de Carnaíba e Socotó, Bahia. Monograph. Instituto de Geociências, Universidade Federal da Bahia, Salvador, 134 pp. Noronha-Oliveira, M.V., Brito-de-Carvalho, C. & Faria, R.G. (2010) Dryadosaura nordestina (NCN). Herpetological Review, 41, 512. Nunes, P.M.S. (2011) Morfologia hemipeniana dos lagartos microteídeos e suas implicações nas relações filogeneticas da família Gymnophthalmidae (Teioidea: Squamata). PhD Thesis, Departamento de Zoologia, Universidade de São Paulo, São Paulo, 137 pp. Nunes, P.M.S., Fouquet, A., Curcio, F.F., Kok, P.J.R. & Rodrigues, M.T. (2012) Cryptic species in Iphisa elegans Gray, 1851 (Squamata: Gymnophitalmidae) revealed by hemipenial morphology and molecular data. Zoological Journal of the Linnean Society, 166, 361–376. http://dx.doi.org/10.1111/j.1096-3642.2012.00846.x Oliveira, B.H.S. (2011) Resultados preliminares sobre o uso do habitat por Anotosaura vanzolinia (Squamata: Gymnophthalmidae) em uma área de Caatinga, Campina Grande, PB. X Congresso de Ecologia do Brasil, 2011, São Lourenço, MG. Palumbi, S.R. (1996) Nucleic acids. II. The polymerase chain reaction. In: Hillis, D.M., Moritz, C. & Mable, B.K. (Eds.), Molecular Systematics. Sinauer & Associates, Sunderland, MA, pp. 205–247. Pellegrino, K.C.M., Rodrigues, M.T. & Yonenaga-Yassuda, Y. (1999a) Chromosomal evolution in the Brazilian lizards of genus Leposoma (Squamata, Gymnophthalmidae) from Amazon and Atlantic rain forests: banding patterns and FISH of telomeric sequences. Hereditas, 131 (1), 15–21. http://dx.doi.org/10.1111/j.1601-5223.1999.00015.x

REDISCOVERY OF ANOTOSAURA COLLARIS Zootaxa 3731 (3) © 2013 Magnolia Press · 367 Pellegrino, K.C.M.;, Rodrigues, M.T. & Yonenaga-Yassuda, Y. (1999b) Chromosomal polymorphisms due to supernumerary chromosomes and pericentric inversions in the eyelidless microteiid lizard Nothobachia ablephara (Squamata, Gymnophthalmidae). Chromosome Research, 7 (4), 247–254. Pellegrino, K.C.M., Rodrigues, M.T., Yonenaga-Yassuda, Y. & Sites, J.W. (2001) A molecular perspective on the evolution of microteiid lizards (Squamata, Gymnophthalmidae), and a new classification for the family. Biological Journal of the Linnean Society, 74, 315–338. http://dx.doi.org/10.1111/j.1095-8312.2001.tb01395.x Pellegrino, K.C.M., Rodrigues, M.T. & Yonenaga-Yassuda, Y. (2003) A triploid karyotype for Leposoma percarinatum (Squamata, Gymnophthalmidae). Journal of Herpetology, 37, 197–199. http://dx.doi.org/10.1670/0022-1511(2003)037[0197:tkolps]2.0.co;2 Peloso, P.L.V., Pellegrino, K.C.M., Rodrigues, M.T. & Ávila-Pires, T.C.S. (2011) Description and phylogenetic relationships of a new genus and species of lizard (Squamata, Gymnophthalmidae) from the Amazonian rainforest of Northern Brazil.American Museum Novitates, 3713, 1–24. http://dx.doi.org/10.1206/3713.2 Pesantes, O.S. (1994) A Method for preparing the hemipenis of preserved snakes. Journal of Herpetology, 28, 93–95. http://dx.doi.org/10.2307/1564686 Peters, J.A. & Donoso-Barros, R. (1970) Catalogue of the Neotropical Squamata: Part II. Lizards and amphisbaenians. United States National Museum Bulletin, 297, 1–293. http://dx.doi.org/10.5479/si.03629236.297.1 Pianka, E. & Vitt, L. (2003) Lizards: Windows to the Evolution of Diversity. University of California Press, Berkeley, 347pp. Posada, D. (2008) jModelTest: Phylogenetic Model Averaging. Molecular Biology and Evolution, 25, 1253–1256. http://dx.doi.org/10.1093/molbev/msn083 Rambaut, A. & Drummond, A.J. (2009) MCMC Trace Analysis Tool Version v1.5.0. Institute of Evolutionary Biology, University of Edinburgh and Department of Computer Science, University of Auckland. Available from: http:// beast.bio.ed.ac.uk/ (accessed 02 October 2013) Rodrigues, M.T. (1986) Uma nova espécie do gênero Phyllopezus de Cabaceiras, Paraiba, Brazil, com comentários sobre a fauna de lagartos da área (Sauria, Gekkonidae). Papéis Avulsos de Zoologia, 36, 237–250. Rodrigues, M.T. (1990) Os Lagartos da Floresta Atlântica Brasileira: Distribuição Atual e Pretérita e suas Implicações para Estudos Futuros. In: Watanabe, S. (Ed.) II Simpósio sobre ecossistemas da costa sul e sudeste brasileira. Academia de Ciências do Estado de São Paulo, São Paulo, pp. 404–410. Rodrigues, M.T. (2003) Herpetofauna da Caatinga. In: Leal, I.R., Tabarelli, M. & Silva, J.M.C. (Eds.), Ecologia e Conservaçao da Caatinga. Universidade Federal de Pernambuco, Recife, pp. 181–236. Rodrigues, M.T., Freire, E.M.X., Pellegrino, K.C.M. & Sites, J.W. (2005) Phylogenetic relationships of a new genus and species of microteiid lizard from the Atlantic forest of north-eastern Brazil (Squamata, Gymnophthalmidae). Zoological Journal of the Linnean Society, 144, 543–557. http://dx.doi.org/10.1111/j.1096-3642.2005.00177.x Rodrigues, M.T., Teixeira Jr., M., Recoder, R.S., Dal Vechio, F., Damasceno, R. & Pellegrino, K.C.M. (2013) A new species of Leposoma (Squamata: Gymnophthalmidae) with four fingers from the Atlantic Forest central corridor in Bahia, Brazil. Zootaxa, 3635, 459–475. Romer, A.S. (1956) Osteology of the Reptiles. The University of Chicago Press, Chicago, 772 pp. Ronquist, F., Teslenko, M., van der Mark, P., Ayres, D.L., Darling, A., Höhna, S., Larget, B., Liu, L., Suchard, M.A. & Huelsenbeck, J.P. (2012) MrBayes 3.2: Efficient Bayesian Phylogenetic Inference and Model Choice across a Large Model Space. Systematic Biology, 31, 1–4. Ruschi, A. (1966) Lista dos répteis do Estado do Espírito Santo. Boletim do Museu de Biologia “Prof. Mello Leitão” (Zoologia), 26, 1–6. Russel, A.P. & Bauer, A.M. (2008) The appendicular locomotor apparatus of Sphenodon and normal-limbed Squamates. In: Gans, C., Gaunt, A.S. & Adler, K. (Eds.), Biology of the Reptilia, Vol. 21. Morphology I. Society for the Study of Amphibians and Reptiles. Academic Press, London, pp. 1–46. Santana, G.G., Vieira, W.L.S., Pereira-Filho, G.A., Delfim, F.R., Lima, Y.C.C. & Vieira, K.S. (2008) Herpetofauna em um fragmento de Floresta Atlântica no Estado da Paraíba, Região Nordeste do Brasil. Biotemas, 21, 75–84. http://dx.doi.org/10.5007/2175-7925.2008v21n1p75 Savage, J.M. (1997) On terminology for the description of the hemipenis of squamate reptiles. Herpetological Journal, 7, 23–25. Sherbrooke, W.C. & Cole, C.J. (1972) Chromosomes of the South American teiid lizard Neusticurus ecpleopus Cope and Neusticurus strangulatus trachodus Uzzell. Copeia, 1972, 886–889. Soares, M. & Caramaschi, U. (1998) A new species of Colobosauroides Cunha, Lima-Verde & Lima, 1991 from the state of Bahia, Brazil (Squamata, Sauria, Gymnophthalmidae). Boletim do Museu Nacional, Nova Série - Zoologia, 388, 1–8. Stamatakis, A. (2006) RAxML-VI-HPC: maximum likelihood-based phylogenetic analyses with thousands of taxa and mixed models. Bioinformatics, 22, 2688–2690. http://dx.doi.org/10.1093/bioinformatics/btl446 Swofford, D.L. (2001) PAUP*. Phylogenetic Analysis Using Parsimony (*and Other Methods) version 4.0 beta 10 for 32-bit Microsoft Windows. Smithsonian Institution: Washington, DC.

368 · Zootaxa 3731 (3) © 2013 Magnolia Press RODRIGUES ET AL. Tamura, K., Peterson, D., Peterson, N., Stecher, G., Nei, M. & Kumar, S. (2011) MEGA5: Molecular Evolutionary Genetics Analysis Using Maximum Likelihood, Evolutionary Distance, and Maximum Parsimony Methods. Molecular Biology and Evolution, 28, 2731–2739. http://dx.doi.org/10.1093/molbev/msr121 Uzzell, T. (1973) A revision of lizards of the genus Prionodactylus, with a new genus for P. leucostictus and notes on the genus Euspondylus (Sauria, Teiidae). Postilla, 159, 1–67. Vanzolini, P. (1976) Two notes on Anotosaura (Sauria, Teiidae). Papéis Avulsos de Zoologia, 30, 119–122. Vanzolini, P.E. & Ramos, A.M.M. (1977) A new species of Colobodactylus, with notes on the distribution of a group of stranded microteiid lizards (Sauria, Gymnophthalmidae). Papéis Avulsos de Zoologia, 31, 19–47. Venables, W.N. & Ripley, B.D. (2002) Modern Applied Statistics with S. Fourth Edition.Springer, New York, 508 pp. Williams, E.E. & Vanzolini, P. (1980) Notes and biogeographic comments on anoles from Brasil. Papéis Avulsos de Zoologia, 34, 99–108. Wiens, J.J., Brandley, M.C. & Reeder, T.W. (2006) Why does a trait evolve multiple times within a clade? Repeated evolution of snakelike body form in squamate reptiles. Evolution, 60, 123–141. http://dx.doi.org/10.1111/j.0014-3820.2006.tb01088.x Yonenaga, Y. (1972) Polimorfismos cromossômicos em roedores brasileiros. PhD Thesis, Instituto de Biociências, Universidade de São Paulo, 214 pp. Yonenaga-Yassuda, Y. & Rodrigues, M.T. (1999) Supernumerary chromosome variation, heteromorphic sex chromosomes and banding patterns in microteiid lizards of the genus Micrablepharus (Squamata, Gymnophthalmidae). Chromosome Research, 7, 21–29. Yonenaga-Yassuda,Y., Vanzolini, P.E., Rodrigues, M.T. & Carvalho, C.M. (1995) Chromosome banding patterns in the unisexual microteiid Gymnophthalmus underwoodi in two related sibling species (Gymnophthalmidae, Sauria). Cytogenetics and Cell Genetics, 70, 29–34. http://dx.doi.org/10.1159/000133985 Yonenaga-Yassuda, Y., Mori, L., Chu, T.H. & Rodrigues, M.T. (1996) Chromosomal banding patterns in the eyelid-less microteiid radiation: Procellosaurinus and Vanzosaura (Squamata, Gymnophthalmidae). Cytogenetics and Cell Genetics, 74, 203–210. http://dx.doi.org/10.1159/000134415 Yonenaga-Yassuda, Y., Rodrigues, M.T. & Pellegrino, K.C.M. (2005) Chromosomal banding patterns in the eyelid-less

microteiid radiation: the X1X2Y:X 1X1X2X2 sex chromosome system in Calyptommatus, and the karyotypes of Psilophthalmus and Tretioscincus (Squamata, Gymnophthalmidae). Genetics and Molecular Biology, 28, 700–709. http://dx.doi.org/10.1590/s1415-47572005000500009 Zaher, H. (1999) Hemipenial morphology of the South American xenodontine snakes, with a proposal for a monophyletic Xenodontinae and a reappraisal of colubroid hemipenes. Bulletin of the American Museum of Natural History, 240, 1–168. Zar, J.H. (2010) Biostatistical Analysis. 5 ed. Prentice Hall, New Jersey, 960 pp.

APPENDIX I. Additional examined material.

Anotosaura vanzolinia—BRAZIL: ALAGOAS: UHE Xingó: MZUSP 80151. BAHIA: Paulo Afonso: RC 004. PERNAMBUCO: Buique: ACG 95, 122, 130; Cabaceiras: MZUSP 45754, 60773, 60775-60777, 60780, 60782-60784, 95304, 95306, 95335. Colobosauroides cearensis—BRAZIL: CEARÁ: Mulungu: MZUSP 79213, 87580-87583; São Gonçalo do Amarante: UFC 3508, 3549, 3556, 3575, 3705. Dryadosaura nordestina—BRAZIL: ALAGOAS: Murici: MZUSP 93219-93222; Passo do Camaragibe: MZUSP 99004- 99006. BAHIA: EEE Wenceslau Guimarães: PEU 041, 081; MTR 22089. PERNAMBUCO: Recife: MNRJ 9931. SERGIPE: PARNA Itabaiana: MTR 16742.

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