Molecular phylogeny, species limits, and biogeography of the Brazilian endemic lizard genus Enyalius (Squamata: Leiosauridae): an example of the historical relationship between Atlantic Forests and Amazonia.

The endemic Brazilian Enyalius encompasses a diverse group of forest lizards with most species restricted to the Atlantic Forest (AF). Their taxonomy is problematic due to extensive variation in color pattern and external morphology. We present the first phylogenetic hypothesis for the genus based on 2102 bp of the mtDNA (cyt-b, ND4, and 16S) and nuclear (c-mos) regions, uncovering all previously admitted taxa (9 spp). Different methods of tree reconstruction were explored with Urostrophus vautieri, Anisolepis grilli and A. longicauda as outgroups. The monophyly of Enyalius and its split into two deeply divergent clades (late Oligocene and early Miocene) is strongly supported. Clade A assembles most lineages restricted to south and southeastern Brazil, and within it Enyalius brasiliensis is polyphyletic; herein full species status of E. brasiliensis and E. boulengeri is resurrected. Clade B unites the Amazonian E. leechii as sister-group to a major clade containing E. bilineatus as sister-group to all remaining species from northeastern Brazil. We detected unrecognized diversity in several populations suggesting putative species. Biogeographical analyses indicate that Enyalius keeps fidelity to shadowed forests, with few cases of dispersal into open regions. Ancient dispersal into the Amazon from an AF ancestor may have occurred through northeastern Brazil.

A new species of Leposoma (Squamata: Gymnophthalmidae) with four fingers from the Atlantic Forest central corridor in Bahia, Brazil.

Leposoma sinepollex sp. nov., a new species of the scincoides group, is described from a mountain region in the Atlantic Forest central corridor in state of Bahia, Brazil. The new species is characterized by elongate dorsal and lanceolate ventral scales arranged in diagonal rows, a single and smooth frontonasal, five supraoculars, absence of pollex, third toe as long as or longer than fourth, absence of striations in lower part of head, parietals longer than wide and as long as interparietal, 27-29 dorsals, 25-29 scales around body, 17-19 ventrals, 12-14 total pores in the male (absent in females), 9-10 and 9-11 subdigital lamellae respectively under IV Finger and IV Toe, and strong sexual color dichromatism with a black pigmentation in the ventral parts of males, creamy in females. The new species is morphologically similar to Leposoma nanodactylus, sharing with it among other features the synapomorphic division of the first supraocular. Phylogenetic analyses of 981 bp of combined sequences (cyt b+ ND4) recovered also a strongly supported (PP = 1,0; BP = 100) sister relationship between both species. The new species and Leposoma nanodactylus are placed sister to all the other Atlantic Forest species, with L. baturitensis being the first to diverge in this radiation. We discuss the distribution of the Atlantic Forest Leposoma, as well as possible scenarios for the origin of the new species.

Diversity of Mycobacteriaceae from aquatic environment at the São Paulo Zoological Park Foundation in Brazil.

We investigated the species diversity of Mycobacteriaceae in surface water samples from six environments at the zoological park in São Paulo, Brazil. Three hundred and eighty isolates were cultivated and identified by phenotypic characteristics (growth rate and pigmentation) and sequencing of hsp65, rpoB and 16S rRNA genes. The results revealed that almost 48% of the isolates could be identified at the species level; about 50% were classified at the genus level, and only less than 2% of the isolates showed an inconclusive identification. The isolates classified at the genus level and not identified were then evaluated by phylogenetic analyses using the same three concatenated target genes. The results allowed us to identify at the genus level some isolates that previously had inconclusive identification, and they also suggested the presence of putative candidate species within the sample, demonstrating that this zoological park is an important source of diversity.

On the distinctiveness of Amapasaurus, its relationship with Loxopholis Cope 1869, and description of a new genus for L. guianensis and L. hoogmoedi (Gymnophthalmoidea/Ecpleopodini: Squamata).

Amapasaurus is a monotypic genus of forest lizards never accessed molecularly and, based on morphological similarities, suggested to be closely related to species of the former Leposoma parietale group Ruibal 1952, currently in the genus Loxopholis Cope 1869. Two other species, formerly allocated in Arthrosaura (A. guianensis and A. hoogmoedi), were tentatively moved to Loxopholis in an extensive molecular revision of Gymnophthalmoidea. Here we add mitochondrial and nuclear sequence data (12S, cyt b, ND4, c-mos and NT3) of Amapasaurus to previously published and new data of all Ecpleopodini genera (except for Adercosaurus), in order to test: i) the close relationship between Amapasaurus and Loxopholis and ii) the position of Loxopholis guianensis and Loxopholis hoogmoedi with three different phylogenetic methods, expanding the knowledge on the current taxonomy of Ecpleopodini. Concatenated analyses of mitochondrial and nuclear data (2303 bp) under Bayesian Inference, Maximum Likelihood and Maximum Parsimony methods recovered a strongly supported sister relationship between Amapasaurus tetradactylus Cunha 1970 and species of Loxopholis. Genetic divergence between Amapasaurus and this assemblage of Loxopholis is high in both mitochondrial (~18% for cyt b) and nuclear (~12% for c-mos) regions, supporting its generic distinctiveness. Differing from the current taxonomy of the Ecpleopodini tribe, our analyses recovered Lo. guianensis and Lo. hoogmoedi as a distinct clade that is sister to all other Loxopholis plus Amapasaurus. Supplemented by external and hemipenial morphology data available from the literature along with DNA sequences, we restrict Loxopholis to the species of the former parietale group of Leposoma and describe a new genus to allocate Lo. guianensis and Lo. hoogmoedi.

Differential staining and microchromosomal variation in karyotypes of four Brazilian species of Tupinambinae lizards (Squamata: Teiidae).

Kayotypes of four neotropical teiid lizard species (Tupinambinae) were herein studied after conventional as well as silver staining and CBG-banding: Crocodilurus amazonicus (2n = 34), Tupinambis teguixin (2n = 36), Tupinambis merianae and Tupinambis quadrilineatus (2n = 38). The karyological data for T. quadrilineatus as well as those obtained using differential staining for all species were unknown until now. The karyotypes of all species presented 12 macrochromosomes identical in morphology, but differed in the number of microchromosomes: 22 in C. amazonicus, 24 in T. teguixin and 26 in T. quadrilineatus and T. merianae. The Ag-NOR located at the secondary constriction at the distal end of pair 2 is shared by all species, contrasting with the variability observed for this character in species of the related Teiinae. CBG-banding revealed a species-specific pattern in T. quadrilineatus with conspicuous interstitial C-blocks at the proximal region of the long arm of pair 4 and the whole heterochromatic short arm of pair 6. The karyological data reported here corroborates the relationship hypothesis obtained for Tupinambis based on molecular characters. T. teguixin presents the putative ancestral karyotype for the genus with 2n = 36 whereas T. merianae and T. quadrilineatus exhibit 2n = 38, due to an additional pair of microchromosomes.

Banding patterns and chromosomal evolution in five species of neotropical Teiinae lizards (Squamata: Teiidae).

Karyotypes of five species of South American teiid lizards from subfamily Teiinae: Ameiva ameiva, Kentropyx calcarata, K. paulensis, K. vanzoi (2n = 50, all acrocentric), and Cnemidophorus ocellifer (2n = 50, all biarmed), are herein described and compared on the basis of conventional and silver staining, and CBG and RBG banding patterns. Meiotic data are also included. Karyotypes of K. paulensis, K. vanzoi, and C. ocellifer are reported here for the first time. Inter-generic variability in Ag-NORs location was detected with NORs occurring at the end of long arm of pair 1 in K. calcarata, K. paulensis, and K. vanzoi; pair 5 in C. ocellifer and pair 7 in A. ameiva. The location of NORs, along with the karyological differences between A. ameiva and the Central American species (A. auberi), corroboretes the molecular-based hypothesis that the genus Ameiva is paraphyletic. Inter-populational heteromorphism in Ag-NORs size was detected between populations of C. ocellifer. RBG and CBG banding data demonstrated that the biarmed condition of the C. ocellifer chromosomes is due to multiple pericentric inversion events instead of addition of constitutive heterochromatin. Differential-staining techniques used here revealed valuable information about Teiinae karyotypic diversity and made it possible to compare these species, contributing to both the better comprehension of their chromosomal evolution and issues on taxa systematics.

Comparative cytogenetics and supernumerary chromosomes in the Brazilian lizard genus Enyalius (Squamata, Polychrotidae).

Cytogenetical analyses based on conventional and differential staining were performed for the first time on five species of the Brazilian lizard genus Enyalius: E. bibronii, E. bilineatus, E. iheringii, E. leechii, and E. perditus. The species share a similar 2n = 36 (12M + 24m) karyotype, comprised of 12 metacentric or submetacentric macrochromosomes, except for an acrocentric pair 6 that characterizes E. bibronii. The 24 microchromosomes were acrocentrics, but in E. perditus two meta/submetacentric microchromosome pairs were unambiguously identified. Karyotypes with 2n = 37 and 2n = 37/38 chromosomes were also observed in some specimens of E. bilineatus as a result of the presence of supernumerary chromosomes (Bs). Ag-NORs were always located at the distal region of the long arm of the submetacentric pair 2. The constitutive heterochromatin was mostly restricted to the pericentromeric regions of some macrochromosomes and microchromosomes. A XX:XY mechanism of sex determination with a dot-like Y microchromosome occurs in E. bilineatus, E. leechii, and E. perditus.

Chromosomal banding patterns in the eyelid-less microteiid lizard radiation: The X1X1X2X2:X1X2Y sex chromosome system in Calyptommatus and the karyotypes of Psilophthalmus and Tretioscincus (Squamata, Gymnophthalmidae)

Cytogenetic studies were performed on five closely related microteiid lizards (Gymnophthalmini), three Calyptommatus species and Psilophthalmus paeminosus from the sand dunes of the middle São Francisco river in the semiarid caatinga of the Brazilian state of Bahia and Tretioscincus oriximinensis from the Brazilian Amazon region. The diploid chromosome number in Calyptommatus species was 2n = 58 in females and 2n = 57 in males due to a multiple X1X1X2X2:X1X2Y sex chromosomes system, while P. paeminosus was 2n = 44 (20M+24m): where M = macrochromosomes and m = microchromosomes) and T. oriximinensis 2n = 42 (18M+24m). A single pair of silver staining nucleolar organizing regions (Ag-NORs) characterizes all five species. Incorporation of 5-BrdU (Bromodeoxyuridine) followed by replication R-banding (RBG) karyotyping allowed the identification of the larger pairs of chromosomes through longitudinal bands and the late replicating regions. Our data reinforce the remarkable chromosomal variability that has been found in the Gymnophthalmidae and the importance of using differential staining for comparative cytogenetics within this group of lizards. Chromosomal evolution in Gymnophthalmini seems to have included chromosomal fission and fusion, pericentric inversions and variation in the amount and localization of constitutive heterochromatin and the Ag-NOR pattern. Different mechanisms of sex determination also evolved independently in this radiation.

Cytogenetic studies in six species of tropiduridae (Sauria)

Cytogenetic studies were performed on six tropidurid lizards: Tropidurus mucujensis, T hispidus, T montanus, T semitaeniatus, T spinulosus and Uranoscodon superciliosus. Conventional and differential (C- and R-bands and Ag-NORs) staining techniques were employed for the characterization of the karyotypes. The analyses were carried out in order to establish the morphological characterization of macro and microchromosomes, the R- and C-banding patterns as well as the silver staining of nucleolus organizer regions (Ag-NORs). R-banding was obtained after in vitro incorporation of 5-bromodeoxyuridine (BrdU). For some species, the banding patterns are described for the first time. All species presented a diploid number of 2n=36, with a basic karyotype constituted by 12 metacentric or submetacentric macrochromosomes and 24 microchromosomes (12M+24m). Although these species present a conservative karyotype, most of them are distinguished by the morphology of the macro and microchromosomes, by location of Ag-NORs, and by amount and distribution of constitutive hetcrochromatin. Sex determination of the XX:XY type was found in T hispidus, T montanusand U. superciliosus. The presence of species specific karyotypes in T spinulosus and U. superciliosus is suggested.