Phylogenetic systematics of dart-poison frogs and their relatives (Amphibia: Athesphatanura: Dendrobatidae)

The known diversity of dart-poison frog species has grown from 70 in the 1960s to 247 at present, with no sign that the discovery of new species will wane in the foreseeable future. Although this growth in knowledge of the diversity of this group has been accompanied by detailed investigations of many aspects of the biology of dendrobatids, their phylogenetic relationships remain poorly understood. This study was designed to test hypotheses of dendrobatid diversification by combining new and prior genotypic and phenotypic evidence in a total evidence analysis. DNA sequences were sampled for five mitochondrial and six nuclear loci (approximately 6,100 base pairs [bp]; x=3,740 bp per terminal; total dataset composed of approximately 1.55 million bp), and 174 phenotypic characters were scored from adult and larval morphology, alkaloid profiles, and behavior. These data were combined with relevant published DNA sequences. Ingroup sampling targeted several previously unsampled species, including Aromobates nocturnus, which was hypothesized previously to be the sister of all other dendrobatids. Undescribed and problematic species were sampled from multiple localities when possible. The final dataset consisted of 414 terminals: 367 ingroup terminals of 156 species and 47 outgroup terminals of 46 species. Direct optimization parsimony analysis of the equally weighted evidence resulted in 25,872 optimal trees. Forty nodes collapse in the strict consensus, with all conflict restricted to conspecific terminals. Dendrobatids were recovered as monophyletic, and their sister group consisted of Crossodactylus, Hylodes, and Megaelosia, recognized herein as Hylodidae. Among outgroup taxa, Centrolenidae was found to be the sister group of all athesphatanurans except Hylidae, Leptodactyidae was polyphyletic, Thoropa was nested within Cycloramphidae, and Ceratophryinae was paraphyletic with respect to Telmatobiinae. Among dendrobatids, the monophyly and content of Mannophryne and Phyllobates were corroborated. Aromobates nocturnus and Colostethus saltuensis were found to be nested within Nephelobates, and Minyobates was paraphyletic and nested within Dendrobates. Colostethus was shown to be rampantly nonmonophyletic, with most species falling into two unrelated cis- and trans-Andean clades. A morphologically and behaviorally diverse clade of median lingual process-possessing species was discovered. In light of these findings and the growth in knowledge of the diversity of this large clade over the past 40 years, we propose a new, monophyletic taxonomy for dendrobatids, recognizing the inclusive clade as a superfamily (Dendrobatoidea) composed of two families (one of which is new), six subfamilies (three new), and 16 genera (four new). Although poisonous frogs did not form a monophyletic group, the three poisonous lineages are all confined to the revised family Dendrobatidae, in keeping with the traditional application of this name. We also propose changes to achieve a monophyletic higher-level taxonomy for the athesphatanuran outgroup taxa. Analysis of character evolution revealed multiple origins of phytotelm-breeding, parental provisioning of nutritive oocytes for larval consumption (larval oophagy), and endotrophy. Available evidence indicates that transport of tadpoles on the dorsum of parent nurse frogs-a dendrobatid synapomorphy-is carried out primitively by male nurse frogs, with three independent origins of female transport and five independent origins of biparental transport. Reproductive amplexus is optimally explained as having been lost in the most recent common ancestor of Dendrobatoidea, with cephalic amplexus arising independently three times. © American Museum of Natural History 2006.

Bathymetric distribution and recruitment of the spider crab Libinia spinosa H. Milne Edwards 1834 in the Ubatuba and Caraguatatuba regions, northern coast of São Paulo, Brazil (Crustacea, Brachyura, Majoidea, Pisidae)

The objective of this study was to investigate the spatial and temporal distribution of Libinia spinosa H. MILNE EDWARDS 1834 on unconsolidated sublittoral bottoms in two regions off the northern coast of the state of São Paulo, and to analyze the influence of environmental factors on the occurrence of this species and observe the recruitment pattern of its young. Crabs were collected monthly (July 2001 through June 2003) at depths of 5, 10, 15, 20, 25, 30 and 35 In, from a fishing boat equipped with two double-rig nets. Samples of water and sediment were collected for analysis of environmental factors. A total of 2112 spider crabs was obtained (701 juveniles and 1411 adults). The highest abundance was observed at depths of 20 and 25 m, in both regions. These localities were characterized by substrate composed of very fine sand and silt-clay. In regard to the temporal distribution, juveniles and adults predominated in the summer and winter months respectively. From these results, one can infer that the distribution of L. spinosa is related to environmental factors favorable for its life cycle; sediment type is the factor which most strongly determines its presence. © E. Schweizerbart'sche Verlagsbuchhandlung (Nägele u. Obermiller), 2007.

Biotic interaction between spionid polychaetes and bouchardiid brachiopods: Paleoecological, taphonomic and evolutionary implications

Shells of Bouchardia rosea (Brachiopoda, Rhynchonelliformea) are abundant in Late Holocene death assemblages of the Ubatuba Bight, Brazil, SW Atlantic. This genus is also known from multiple localities in the Cenozoic fossil record of South America. A total of 1211 valves of B. rosea, 2086 shells of sympatric bivalve mollusks (14 nearshore localities ranging in depth from 0 to 30 m), 80 shells of Bouchardia zitteli, San Julián Formation, Paleogene, Argentina, and 135 shells of Bouchardia transplatina, Camacho Formation, Neogene, Uruguay were examined for bioerosion traces. All examined bouchardiid shells represent shallow-water, subtropical marine settings. Out of 1211 brachiopod shells of B. rosea, 1201 represent dead individuals. A total of 149 dead specimens displayed polychaete traces (Caulostrepsis). Live polychaetes were found inside Caulostrepsis borings in 10 life-collected brachiopods, indicating a syn-vivo interaction (Caulostrepsis traces in dead shells of B. rosea were always empty). The long and coiled peristomial palps, large chaetae on both sides of the 5th segment, and flanged pygidium found in the polychaetes are characteristic of the polychaete genus Polydora (Spionidae). The fact that 100% of the Caulostrepsis found in living brachiopods were still inhabited by the trace-making spionids, whereas none was found in dead hosts, implies active biotic interaction between the two living organisms rather than colonization of dead brachiopod shells. The absence of blisters, the lack of valve/site stereotypy, and the fact that tubes open only externally are all suggestive of a commensal relationship. These data document a new host group (bouchardiid rhynchonelliform brachiopods) with which spionids can interact (interestingly, spionid-infested sympatric bivalves have not been found in the study area despite extensive sampling). The syn-vivo interaction indicates that substantial bioerosion may occur when the host is alive. Thus, the presence of such bioerosion traces on fossil shells need not imply a prolonged post-mortem exposure of shells on the sea floor. Also, none of the Paleogene and Neogene Bouchardia species included any ichnological evidence for spionid infestation. This indicates that the Spionidae/ Bouchardia association may be geologically young, although the lack of older records may also reflect limited sampling and/or taphonomic biases.

Polimorfismo nos genes metilenotetrahidrofolato redutase e cistationina-beta-sintase e a sua relação com eventos vaso-oclusivos na doença falciforme

Sickle cell disease is an inflammatory condition with a pathophysiology that involves vaso-occlusive episodes. Mutations of the methylenetetrahydrofolate reductase (MTHFR) and cystathionine beta-synthase (CBS) genes are risk factors for vascular disease. Due to the importance of identifying risk factors for vaso-occlusive events in sickle cell patients, we investigated the frequencies of the C677T and 844ins68 mutations of the MTHFR and CBS genes, respectively. Three hundred patients with Hb SS, HB SC and HbS/Beta thalassemia, from Brasília, Goiânia, Rio de Janeiro, São Jose do Rio Preto and São Paulo were evaluated. Samples of 5 mL of venous blood were collected in EDTA after informed consent was received from patients. Classical diagnostic methods were used to confirm the hemoglobin phenotypes. The hemoglobin genotypes and polymorphisms studied were evaluated by Restriction Fragment Length Polymorphism and Allele Specific amplification. The results showed that 93 patients (31.00%) were heterozygous and 13 (4.33%) homozygous for the C677T mutation and 90 were heterozygotes (30.00%) and 8 homozygous (2.66%) for the 844ins68 mutation, both with significant differences for genotype frequency between the localities. The allelic frequencies are in Hardy-Weinberg equilibrium for both polymorphisms. The frequency of mutations was significant and the presence of related vaso-occlusive events was more common in patients with Hb SS (p = 0007). The 844ins68 mutation was approximately three times more frequent in patients with vaso-occlusive complications (p = 0011). The C677T mutation did not prove to be associated with risk of vaso-occlusive events (p = 0.193). A C677T-844ins68 interaction occurred in 12.08% of the patients, doubling the risk of vaso-occlusive manifestations. The frequencies of the polymorphisms are consistent with those expected in the Brazilian population. The presence of the 844ins68 mutation of the CBS gene proved to be a potential risk factor for vaso-occlusive events in sickle cell patients.

Proposição e validações de equações para estimativa da erosividade de dois municípios do estado de São Paulo

The equations and extrapolation use to localities whose characteristics of soil and climate, even if partial, distinguish the town to which they were generated, still permeate in studies to estimate the rainfall erosivity (EI 30). This work has objective to propose and validate mathematical equations to estimate the rainfall erosivity of two cities of Sao Paulo State's. The adjusted to estimate obtaining and validate data of equations of erosivity (EI 30) according to values of coefficient of rain (Rc) were obtained from pluviographic and pluviometric rainfall data, respectively, using of distinct historical rainfall series. Mutiple comparisions test and confidence intervals were performed to compare absolute average of EI 30, pluviometric data (Pp), and Rc. The correlation between EI 30 and Rc was verified by of Pearson correlation coefficient. Test of the hypothesis of equality between population variance was used to compare the equations. Pluviometrics data of historical series rainfall data different than those that the models were generated were used to validate and to assess the performance of the equations, proposed of this study and compare them with another equation already consolidated in literature. The results show that for the conditions under which the study was conducted, the simple linear equations, shown to be the most appropriate to estimate the rainfall erosivity these two cities. According to the test of the hypothesis of equality variances between populations, the equations adjusted for each city differ statistically so that the rainfall erosivity of each city must be estimated by their respective equation.

Climatic variables and altitude as predictors of anuran species richness and number of reproductive modes in Brazil

Because anuran species are highly dependent on environmental variables, we hypothesized that anuran species richness and the number of reproductive modes from different Brazilian localities vary according to climatic and altitudinal variables. Published data were compiled from 36 Brazilian localities and climatic and altitudinal data were extracted from an available database. A partial redundancy analysis (pRDA) showed that 23.5% of the data set's variation was explained by climatic and altitudinal data, while the remaining 76.5% remained unexplained. This analysis suggests that other factors not analysed herein may also be important for predicting anuran species richness and the number of reproductive modes in Brazil. Altitude and total annual rainfall were positively correlated with anuran species richness and the number of reproductive modes, and total annual rainfall was strongly associated with these two biotic variables in the triplot of pRDA. The positive association of total annual rainfall and the negative association of the concentration of annual rainfall were already expected based on physiological and reproductive requirements of anurans. On the other hand, temperature was not associated with richness or the number of reproductive modes. Copyright © 2010 Cambridge University Press.

Amphibia, Anura, Centrolenidae, Hyalinobatrachium carlesvilai Castroviejo-Fisher, Padial, Chaparro, Aguayo and de la Riva, 2009: First country record, Brazil

We present the first Brazilian record of the recently-described glassfrog Hyalinobatrachium carlesvilai Castroviejo-Fisher, Padial, Chaparro, Aguayo and de la Riva, 2009. This species, previously known in Peru and Bolivia, was collected at two localities at the municipality of Aripuanã, northern state of Mato Grosso, Brazil. © 2010 Check List and Authors.

The lauraceae collected in brazil by ludwig riedel - I

The species of Lauraceae collected by Ludwig Riedel, excluding Ocotea, are commented and contex-tualized according to the different itineraries he travelled in Brazil, from 1821 to 1836, indicating the localities and dates of collections, whenever possible. To complete this work, two diaries of Riedel have been consulted and the information from them cross-checked with the annotations found on specimen labels. An annotated list of 48 entries provides information on the status of species names and their types, currently accepted names, and collections found in different herbaria. Lectotypes for five names, Aydendron verrucosum var. attenuata, Aydendron parviflorum, Mespilodaphne parviflora, Nectandra spicata, and Oreodaphne fasciculata, are designated. The species of Ocotea collected by Riedel will be treated in part II of this series. © President and Fellows of Harvard College, 2012.

The lauraceae collected in brazil by ludwig riedel - II. Ocotea

The species of Ocotea (Lauraceae) collected by Ludwig Riedel are commented and contextualized according to the different itineraries he travelled in Brazil, from 1821 to 1836, indicating the localities and dates of collections, whenever possible. To complete this work, two diaries of Riedel have been consulted and the information from them crossed-checked with the annotations found on specimen labels. An annotated list of 44 entries provides information on the status of species names and their types, currently accepted names, and collections found in different herbaria. Lectotypes for 19 names are designated, two new synonyms are proposed, as well as the new combination Ocotea estrellensis (Meisn.) P.L.R. Moraes, based on Oreodaphne estrellensis Meisn. © President and Fellows of Harvard College, 2012.

From widespread to microendemic: Molecular and acoustic analyses show that Ischnocnema guentheri (Amphibia: Brachycephalidae) is endemic to Rio de Janeiro, Brazil

Many species of tropical amphibians are restricted to very small ranges, and this microendemism coupled with ongoing habitat loss and susceptibility to emerging pathogens imperils the long-term persistence of these species. Incomplete taxonomic and distributional knowledge may obscure conservation assessment, particularly in putatively widespread species that are typically considered to be of Least Concern in Red List assessments, but that in fact may constitute complexes of partly microendemic species. Such is the case in the Steindachner's Robber Frog, Ischnocnema guentheri which, together with the recently recognized Ischnocnema henselii, is thought to occupy most of the Brazilian Atlantic Forest. To test whether these taxa may constitute a species complex of range-restricted and thus potentially threatened species, we analyzed 160 samples of I. guentheri and/or I. henselii for two molecular markers, 16S rRNA (16S) and recombination activation gene 1 (RAG1). To verify the monophyly of the complex, closely related species were also included in the 16S analysis. Congruent evidence from the molecular data and from analyses of advertisement calls support the existence of six distinct species within the complex: I. guentheri and I. henselii as well as four candidate new species. The lineages are distributed as a mosaic in the Atlantic Forest and are sympatric at some localities without indication of admixture. Their phylogeographical pattern partially agrees with paleo-models for the Atlantic Forest, but also suggests the existence of micro-refugia in less stable areas. I. guentheri, previously considered to be widespread, was found only in its type locality, a reserve within the urban area of Rio de Janeiro city. Although none of the species studied appears highly threatened with extinction, we recommend their IUCN threat status to be re-evaluated carefully for the next comprehensive update of the Red List of Brazil's amphibians. © 2013 Springer Science+Business Media Dordrecht.

Comparison of the population biology of Epialtus bituberculatus from two rocky shores with distinct hydrodynamic patterns

The population biology of Epialtus bituberculatus was compared between two different intertidal localities with differing levels of wave exposure. Sampling was conducted monthly between January and December, 2001 on seaweed banks of Sargassum cymosum in the intertidal zone of the rocky shores Grande (GR) (23°23′S-45°03′W) and Domingas Dias (DD) (23°29′S-45°08′W). Four hundred and twenty-eight crabs were captured at the GR site: 111 juvenile males, 106 adult males, 57 juvenile females, 17 adult females and 137 ovigerous females; while 455 specimens were obtained at the DD site: 76 juvenile males, 113 adult males, 37 juvenile females, 40 adult females and 189 ovigerous females. The population from GR showed a non-normal distribution and from DD a normal distribution. The sex-ratio (female/male) was 1:0.97 at GR (χ2 = 0.77, P = 0.084), whereas it was 1:1.41 at DD (χ2 = 13.03, P < 0.001). The largest individuals occurred at DD (U = 78249.0, P < 0.001). The estimated size at sexual maturity was 6.3 and 5.0 mm carapace width (CW) for males, and 5.4 and 5.2 mm CW for females, from GR and DD, respectively. The observed differences in E. bituberculatus between the studied localities might be explained by the different degrees of wave exposure between sites. However, other factors that might also explain the observed differences (e.g. temperature, salinity and food availability) cannot be discarded as relevant in influencing the population structure between sites herein studied. © 2012 Marine Biological Association of the United Kingdom.

Systematics of spiny-backed treefrogs (Hylidae: Osteocephalus): An Amazonian puzzle

Spiny-backed tree frogs of the genus Osteocephalus are conspicuous components of the tropical wet forests of the Amazon and the Guiana Shield. Here, we revise the phylogenetic relationships of Osteocephalus and its sister group Tepuihyla, using up to 6134 bp of DNA sequences of nine mitochondrial and one nuclear gene for 338 specimens from eight countries and 218 localities, representing 89% of the 28 currently recognized nominal species. Our phylogenetic analyses reveal (i) the paraphyly of Osteocephalus with respect to Tepuihyla, (ii) the placement of 'Hyla' warreni as sister to Tepuihyla, (iii) the non-monophyly of several currently recognized species within Osteocephalus and (iv) the presence of low (<1%) and overlapping genetic distances among phenotypically well-characterized nominal species (e.g. O. taurinus and O. oophagus) for the 16S gene fragment used in amphibian DNA barcoding. We propose a new taxonomy, securing the monophyly of Osteocephalus and Tepuihyla by rearranging and redefining the content of both genera and also erect a new genus for the sister group of Osteocephalus. The colouration of newly metamorphosed individuals is proposed as a morphological synapomorphy for Osteocephalus. We recognize and define five monophyletic species groups within Osteocephalus, synonymize three species of Osteocephalus (O. germani, O. phasmatus and O. vilmae) and three species of Tepuihyla (T. celsae, T. galani and T. talbergae) and reallocate three species (Hyla helenae to Osteocephalus, O. exophthalmus to Tepuihyla and O. pearsoni to Dryaderces gen. n.). Furthermore, we flag nine putative new species (an increase to 138% of the current diversity). We conclude that species numbers are largely underestimated, with most hidden diversity centred on widespread and polymorphic nominal species. The evolutionary origin of breeding strategies within Osteocephalus is discussed in the light of this new phylogenetic hypothesis, and a novel type of amplexus (gular amplexus) is described. © 2013 The Norwegian Academy of Science and Letters.

Rediscovery and re-description of Ischnocnema nigriventris (Lutz, 1925) (Anura: Terrarana: Brachycephalidae)

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

Cytogenetic analysis of Phyllomedusa distincta Lutz, 1950 (2n = 2x = 26), P. tetraploidea Pombal and Haddad, 1992 (2n = 4x = 52), and their natural triploid hybrids (2n = 3x = 39) (Anura, Hylidae, Phyllomedusinae)

Background: Natural polyploidy has played an important role during the speciation and evolution of vertebrates, including anurans, with more than 55 described cases. The species of the Phyllomedusa burmeisteri group are mostly characterized by having 26 chromosomes, but a karyotype with 52 chromosomes was described in P. tetraploidea. This species was found in sintopy with P. distincta in two localities of São Paulo State (Brazil), where triploid animals also occur, as consequence of natural hybridisation. We analyse the chromosomes of P. distincta, P. tetraploidea, and their triploid hybrids, to enlighten the origin of polyploidy and to obtain some evidence on diploidisation of tetraploid karyotype.Results: Phyllomedusa distincta was 2n = 2x = 26, whereas P. tetraploidea was 2n = 4x = 52, and the hybrid individuals was 2n = 3x = 39. In meiotic phases, bivalents were observed in the diploid males, whereas both bivalents and tetravalents were observed in the tetraploid males. Univalents, bivalents or trivalents; metaphase II cells carrying variable number of chromosomes; and spermatids were detected in the testis preparations of the triploid males, indicating that the triploids were not completely sterile. In natural and experimental conditions, the triploids cross with the parental species, producing abnormal egg clutches and tadpoles with malformations. The embryos and tadpoles exhibited intraindividual karyotype variability and all of the metaphases contained abnormal constitutions. Multiple NORs, detected by Ag-impregnation and FISH with an rDNA probe, were observed on chromosome 1 in the three karyotypic forms; and, additionally, on chromosome 9 in the diploids, mostly on chromosome 8 in the tetraploids, and on both chromosome 8 and 9 in the triploids. Nevertheless, NOR-bearing chromosome 9 was detected in the tetraploids, and chromosome 9 carried active or inactive NORs in the triploids. C-banding, base-specific fluorochrome stainings with CMA3 and DAPI, FISH with a telomeric probe, and BrdU incorporation in DNA showed nearly equivalent patterns in the karyotypes of P. distincta, P. tetraploidea, and the triploid hybrids.Conclusions: All the used cytogenetic techniques have provided strong evidence that the process of diploidisation, an essential step for stabilising the selective advantages produced by polyploidisation, is under way in distinct quartets of the tetraploid karyotype. © 2013 Gruber et al.; licensee BioMed Central Ltd.

Genetic Structure of Lutzomyia longipalpis Populations in Mato Grosso Do Sul, Brazil, Based on Microsatellite Markers

Background:Lutzomyia longipalpis (Diptera: Psychodidae) is the major vector of Leishmania (Leishmania) infantum and thus plays a crucial role in the epidemiology of American visceral leishmaniasis (AVL). This vector is the best studied species of sand fly in the Neotropical region. Many studies claim that this vector is in fact a species complex; however there is still no consensus regarding the number of species that belong into this complex or the geographical distribution of sibling species. The aim of the present study was to analyze the genetic relationships within Lu. longipalpis populations in the state of Mato Grosso do Sul (MS), Brazil.Methodology/Principal Findings:We collected 30 Lu. longipalpis (15 females and 15 males) from five localities (Campo Grande, Três Lagoas, Aquidauana, Miranda and Bonito) and 30 Lu. Cruzi from Corumbá, totaling 180 sandflies from MS, and 30 Lu. longipalpis from Estrela de Alagoas, state of Alagoas (AL), Northeast Brazil. We show that eight previously described microsatellite loci were sufficient in distinguishing Lu. longipalpis from Lu. Cruzi, which is a closely related species, and in differentiating between Lu. longipalpis collected in MS versus Estrela de Alagoas. Analyses of the genotypes revealed introgression between sympatric Lu. longipalpis and Lu. Cruzi.Conclusions/Significance:Our findings support the hypothesis of cryptic species within the Lu. longipalpis complex. Furthermore, our data revealed introgression between Lu. longipalpis and Lu. cruzi. This phenomenon should be further investigated to determine the level and incidence of hybridization between these two species. We also demonstrated that microsatellite markers are a powerful tool for differentiating sand fly populations and species. The present study has elucidated the population structure of Lu. longipalpis in MS and, by extension, the Neotropical Lu. longipalpis complex itself. © 2013 Santos et al.