Taxonomy of the Threadsnakes of the tribe Epictini (Squamata: Serpentes: Leptotyphlopidae) in Colombia

Threadsnakes of the tribe Epictini are endemic to the New World, occurring from the United States to Argentina, mostly in the Neotropical region. Currently, the taxonomic status of most species is unclear and there has been no previous attempt of a comprehensive taxonomic revision of Neotropical taxa. Taxonomy of the group is a difficult task due to the paucity of geographic samples, general homogeneous morphology and brevity of species descriptions. Therefore, the only way to address the taxonomic status of existing names is through detailed characterization of the types and the search for additional material of the poorly known species. In this study, we evaluated the taxonomic status of the Colombian threadsnakes and report on geographical variation of meristic, morphometric, colour pattern, and hemipenis characters. On the basis of available samples we recognize the following species in Colombia: Epictia goudotii, E. magnamaculata, E. signata, Rena nicefori, Tricheilostoma brevissimum, T. dugandi, T. joshuai and T. macrolepis. We discuss the systematic position of Rena nicefori and propose its allocation in the genus Tricheilostoma based on a unique combination of morphological characters. Furthermore, we provide a key to the representatives of the tribe Epictini in Colombia.

Second and Third Order Ionospheric Effects on GNSS Positioning: A Case Study in Brazil

The Global Positioning System (GPS) transmits signals in two frequencies. It allows the correction of the first order ionospheric effect by using the ionosphere free combination. However, the second and third order ionospheric effects, which combined may cause errors of the order of centimeters in the GPS measurements, still remain. In this paper the second and third order ionospheric effects, which were taken into account in the GPS data processing in the Brazilian region, were investigated. The corrected and not corrected GPS data from these effects were processed in the relative and precise point positioning (PPP) approaches, respectively, using Bernese V5.0 software and the PPP software (GPSPPP) from NRCAN (Natural Resources Canada). The second and third order corrections were applied in the GPS data using an in-house software that is capable of reading a RINEX file and applying the corrections to the GPS observables, creating a corrected RINEX file. For the relative processing case, a Brazilian network with long baselines was processed in a daily solution considering a period of approximately one year. For the PPP case, the processing was accomplished using data collected by the IGS FORT station considering the period from 2001 to 2006 and a seasonal analysis was carried out, showing a semi-annual and an annual variation in the vertical component. In addition, a geographical variation analysis in the PPP for the Brazilian region has confirmed that the equatorial regions are more affected by the second and third order ionospheric effects than other regions.

Vertebrate dispersal syndromes along the Atlantic forest: broad-scale patterns and macroecological correlates

Aim To assess the geographical variation in the relative importance of vertebrates, and more specifically of birds and mammals, as seed dispersal agents in forest communities, and to evaluate the influence of geographical and climatic factors on the observed trends.Location One hundred and thirty-five forest communities in the Brazilian Atlantic forest.Methods We collected data on dispersal modes for 2292 woody species. By combining species x site with species x trait matrices, we obtained the percentages of endozoochory, ornithochory, mastozoochory and the mean fruit diameter for the local forest communities. We used Spearman's correlation to assess bivariate relationships between variables. Subsequently, we performed paired t-tests to verify if variations in frequency of dispersal modes and mean fruit diameter were influenced by altitude or temperature. Then, we applied multiple linear regressions to evaluate the effect of geographical and climatic variables on variation in the relative frequency of dispersal modes and mean fruit diameter across communities.Results We found no consistent latitudinal or longitudinal trend in the percentage of vertebrate-dispersed species, neither bird- nor mammal-dispersed species along the Atlantic forest. Endozoochory was affected chiefly by annual mean rainfall, increasing towards moister sites. Forest communities located at higher altitudes had a higher percentage of bird-dispersed species. Even when sites with identical values of annual mean temperature were compared, altitude had a positive effect on ornithochory. Conversely, we found a higher percentage of mammal-dispersed species in warmer forests, even when locations at the same altitudinal belts were contrasted. Fruit diameter was clearly related to altitude, decreasing towards higher elevations.Main conclusions This is the first analysis of a large data set on dispersal syndromes in tropical forest communities. Our findings support the hypotheses that: (1) geographical variation in the relative number of fleshy fruit species is mainly driven by moisture conditions and is relatively independent of geographical location, and (2) broad-scale trends in fruit size correspond to geographical variation in the relative importance of mammals and birds as seed dispersal agents at the community level.

Genetic diversity in natural populations of Piper cernuum

Piper cernuum is a native plant of the Brazilian Atlantic rain forest. This work studies the distribution of allozyme diversity in P. cernuum natural populations in order to establish a strategy for sustainable management and conservation. Leaf samples were collected in two Brazilian states. High divergences among populations (F-SR = 0.380) and low divergences among regions (F-RT = -0.069) and among gaps of the same population (F-GT = 0.062) were found. No association between the geographical variation and the genetic distance was detected. An excess of heterozygotes was detected in the populations (F-IS = -0.170), suggesting selection in favor of heterozygotes. The results, and the fact that the species depends on constant gap formation for maintenance of its dynamism, suggest that the founder effect is largely responsible for the structuring of populations. For sustainable management, the maintenance of plants/reproductive branches in the gaps is of major importance. The genotypes produced in these gaps are responsible for the establishment of new gaps and are the foundation for new populations, maintaining the dynamics of allele movement.

A Simple Method Based on ICP-MS for Estimation of Background Levels of Arsenic, Cadmium, Copper, Manganese, Nickel, Lead, and Selenium in Blood of the Brazilian Population

Throughout the world, biomonitoring has become the standard for assessing exposure of individuals to toxic elements as well as for responding to serious environmental public health problems. However, extensive biomonitoring surveys require rapid and simple analytical methods. Thus, a simple and high-throughput method is proposed for the determination of arsenic (As), cadmium (Cd), copper (Cu), manganese (Mn), nickel (Ni), lead (Pb), and selenium (Se) in blood samples by using inductively coupled plasma-mass spectrometry (ICP-MS). Prior to analysis, 200 l of blood samples was mixed with 500 l of 10% v/v tetramethylammonium hydroxide (TMAH) solution, incubated for 10 min, and subsequently diluted to 10 ml with a solution containing 0.05% w/v ethylenediamine tetraacetic acid (EDTA) + 0.005% v/v Triton X-100. After that, samples were directly analyzed by ICP-MS (ELAN DRC II). Rhodium was selected as an internal standard with matrix-matching calibration. Method detection limits were 0.08, 0.04, 0.5, 0.09, 0.12, 0.04, and 0.1 g//L for As, Cd, Cu, Mn, Ni, Pb, and Se, respectively. Validation data are provided based on the analysis of blood samples from the trace elements inter-\comparison program operated by the Institut National de Sante Publique du Quebec, Canada. Additional validation was provided by the analysis of human blood samples by the proposed method and by using electrothermal atomic absorption spectrometry (ETAAS). The method was subsequently applied for the estimation of background metal blood values in the Brazilian population. In general, the mean concentrations of As, Cd, Cu, Mn, Ni, Pb, and Se in blood were 1.1, 0.4, 890, 9.6, 2.1, 65.4, and 89.3 g/L, respectively, and are in agreement with other global populations. Influences of age, gender, smoking habits, alcohol consumption, and geographical variation on the values were also considered. Smoking habits influenced the levels of Cd in blood. The levels of Cu, Mn, and Pb were significantly correlated with gender, whereas Cu and Pb were significantly correlated with age. There were also interesting differences in Mn and Se levels in the population living in the north of Brazil compared to the south.

Antiviral activity of plants occurring in the state of minas gerais (Brazil): Part III

A total of 24 extracts from 14 plant species collected at the state of Minas Gerais, Brazil, and belonging to five botanical families (Annonaceae, Apocynaceae, Ochnaceae, Polygonaceae and Vitaceae) was screened for cytotoxicity in cultured Vero cells and for antiviral activity against human herpes virus type 1 (HSV-1), vaccinia virus (VACV) and murine encephalomyocarditis virus (EMCV). The highest cytotoxicity (CC 50 < 10 μg/mL) was observed for the ethanol extracts from Annona coriacea fruits and seeds. Extracts from Hancornia speciosa, Ouratea castaneafolia and O. semisrrata were the only ones that have shown activity against all the three viruses assayed. Extracts from Polygonum spectabile, Hancornia speciosa, Himatanthus phagedaenica, Ouratea spectabilis and O. semiserrata were the most active against HSV-1 (EC 50 < 50 mg/mL), with favorable SI values (8.0 to 10.0). Hancornia speciosa and Anaxagorea dolichocarpa were the most active against EMCV (EC 50 50 - 100 μg/mL), with reasonable SI values (5.2 to 6.1), while moderate to low activity (EC 50 > 100 μg/mL) was observed for Ouratea spectabilis and O. semiserrata. A total of 7 plant species, Ouratea semiserrata, O. spectabilis, O. castanaeafolia, Rollinia laurifolia, Cissus erosa, Polygonum spectabile, and Hancornia speciosa, were active against VACV, disclosing EC50 < 50 μg/mL and SI values ranging from 6.6 to 67.3. In total, 10 out of the 14 species were selected from a literature survey on plants used to treat viral diseases in Brazil; these species were responsible for 70% of the positive results.

Mudanças ontogenéticas na dieta de Oreochromis niloticus and Tilapia rendalli da represa de Barra Bonita (rio Tietê, Estado de São Paulo, Brasil)

The Nile Tilapia, Oreochromis niloticus, and the Congo Tilapia, Tilapia rendalli, are important members of the African cichlids, and have been introduced to many Brazilian lakes and reservoirs. These species exhibit a large feeding flexibility and may modify their habits during their growth. In the Barra Bonita reservoir, these species are well adapted, representing more than 80% of fish. This study aimed to analyze ontogenetic variation with regard to the diet of these species in this important reservoir. Samples were taken monthly, from March 2007 to February 2008, in Anhembi, São Paulo State. Both species were analyzed by grouping individuals according to size classes. The coexistence of these species was observed in this environment, to which fish were introduced, as well as discreet differences in diet, being that Oreochromis niloticus was considered as an detritivorous, since the detritus was constant in the diet of almost all size classes, and presents some changes in its diet according to the different size classes. While T. rendalli may was defined as herbivorous, and the contribution of food resources to the diet of T. rendalli seems to be different from that of O. niloticus along the size classes.

Development of microsatellite markers for the Neotropical endemic Brazilian Guanabara frog, Euparkerella brasiliensis, through 454 shotgun pyrosequencing

The new-generation 454 GS-FLX Titanium pyrosequencing was used to isolate microsatellite markers for the Brazilian Guanabara frog, Euparkerella brasiliensis, an Atlantic forest endemic species. Three multiplex polymerase chain reaction sets were optimized for genotyping of 11 polymorphic (di- and tetranucleotide) microsatellite markers. Genetic diversity was assessed in 21 individuals from a population (Reserva Ecológica de Guapiaçu, REGUA) locatedin the central region of the Rio de Janeiro State, in Brazil. The mean number of alleles per locus ranged from 3 to 12. Observed and expected heterozygosities ranged from 0.095 to 0.905 and from 0.094 to 0.904, respectively. After using the Bonferroni correction for multiple tests, there was no evidence of linkage disequilibrium between pairs of loci but deviations for Hardy-Weinberg equilibrium were found in 4 loci. We found no evidence for allele dropouts or stuttering, but we detected the presence of null alleles at loci Eb10 and Eb36. These markers will be useful for analyses of fine-scale population structure and determination of relative effects of habitat loss and fragmentation on population genetic variability within species. © FUNPEC-RP.

Circulation of Different Lineages of Dengue Virus 2, Genotype American/Asian in Brazil: Dynamics and Molecular and Phylogenetic Characterization

The American/Asian genotype of Dengue virus type 2 (DENV-2) was introduced into the Americas in the 80′s. Although there is no data showing when this genotype was first introduced into Brazil, it was first detected in Brazil in 1990. After which the virus spread throughout the country and major epidemics occurred in 1998, 2007/08 and 2010. In this study we sequenced 12 DENV-2 genomes obtained from serum samples of patients with dengue fever residing in São José do Rio Preto, São Paulo (SJRP/SP), Brazil, in 2008. The whole open reading frame or envelope sequences were used to perform phylogenetic, phylogeographic and evolutionary analyses. Isolates from SJRP/SP were grouped within one lineage (BR3) close to isolates from Rio de Janeiro, Brazil. Isolates from SJRP were probably introduced there at least in 2007, prior to its detection in the 2008 outbreak. DENV-2 circulation in Brazil is characterized by the introduction, displacement and circulation of three well-defined lineages in different times, most probably from the Caribbean. Thirty-seven unique amino acid substitutions were observed among the lineages, including seven amino acid differences in domains I to III of the envelope protein. Moreover, we dated here, for the first time, the introduction of American/Asian genotype into Brazil (lineage BR1) to 1988/89, followed by the introduction of lineages BR2 (1998-2000) and BR3 (2003-05). Our results show a delay between the introduction and detection of DENV-2 lineages in Brazil, reinforcing the importance and need for surveillance programs to detect and trace the evolution of these viruses. Additionally, Brazilian DENV-2 differed in genetic diversity, date of introduction and geographic origin and distribution in Brazil, and these are important factors for the evolution, dynamics and control of dengue. © 2013 Drumond et al.

Comparative cytogenetic analysis of some species of the Dendropsophus microcephalus group (Anura, Hylidae) in the light of phylogenetic inferences

Background: Dendropsophus is a monophyletic anuran genus with a diploid number of 30 chromosomes as an important synapomorphy. However, the internal phylogenetic relationships of this genus are poorly understood. Interestingly, an intriguing interspecific variation in the telocentric chromosome number has been useful in species identification. To address certain uncertainties related to one of the species groups of Dendropsophus, the D. microcephalus group, we carried out a cytogenetic analysis combined with phylogenetic inferences based on mitochondrial sequences, which aimed to aid in the analysis of chromosomal characters. Populations of Dendropsophus nanus, Dendropsophus walfordi, Dendropsophus sanborni, Dendropsophus jimi and Dendropsophus elianeae, ranging from the extreme south to the north of Brazil, were cytogenetically compared. A mitochondrial region of the ribosomal 12S gene from these populations, as well as from 30 other species of Dendropsophus, was used for the phylogenetic inferences. Phylogenetic relationships were inferred using maximum parsimony and Bayesian analyses.Results: The species D. nanus and D. walfordi exhibited identical karyotypes (2n = 30; FN = 52), with four pairs of telocentric chromosomes and a NOR located on metacentric chromosome pair 13. In all of the phylogenetic hypotheses, the paraphyly of D. nanus and D. walfordi was inferred. D. sanborni from Botucatu-SP and Torres-RS showed the same karyotype as D. jimi, with 5 pairs of telocentric chromosomes (2n = 30; FN = 50) and a terminal NOR in the long arm of the telocentric chromosome pair 12. Despite their karyotypic similarity, these species were not found to compose a monophyletic group. Finally, the phylogenetic and cytogenetic analyses did not cluster the specimens of D. elianeae according to their geographical occurrence or recognized morphotypes.Conclusions: We suggest that a taxonomic revision of the taxa D. nanus and D. walfordi is quite necessary. We also observe that the number of telocentric chromosomes is useful to distinguish among valid species in some cases, although it is unchanged in species that are not necessarily closely related phylogenetically. Therefore, inferences based on this chromosomal character must be made with caution; a proper evolutionary analysis of the karyotypic variation in Dendropsophus depends on further characterization of the telocentric chromosomes found in this group. © 2013 Medeiros et al.; licensee BioMed Central Ltd.

Drivers of clutch-size in Fork-tailed Flycatchers (Tyrannus savana) at temperate and tropical latitudes in South America

Many species of birds exhibit a latitudinal gradient in annual reproductive investment, laying more eggs and producing more nestlings at higher latitudes. However, few studies have evaluated the mechanisms that underlie such patterns and such differences in grassland birds specifically. We monitored nests of Fork-tailed Flycatchers (Tyrannus savana) over two breeding seasons at a tropical site in Bolivia (in 2010-11 and 2011-12) and three breeding seasons at a southern temperate site in Argentina (2010-11, 2011-12 and 2012-13), testing two hypotheses explaining variation in clutch-size among populations: the food-limitation hypothesis and the nest-predation hypothesis. Mean clutch-size and mean brood-size were significantly larger at the temperate study site than at the tropical site. Availability of arthropod food per individual bird was significantly higher at the temperate site. There was no relationship, positive or negative, between rates of nest predation and either clutch- or brood-size, and thus no support for the nest-predation hypothesis. We conclude that food availability explains much of the latitudinal variation in clutch-size in this species. We discuss avenues for future research on the mechanisms underlying geographical variation in the life histories of Neotropical birds.

Two periods of total testicular regression are peculiar events of the annual reproductive cycle of the black Myotis bat, Myotis nigricans (Chiroptera: Vespertilionidae)

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

A simple method based on ICP-MS for estimation of background levels of arsenic, cadmium, copper, manganese, nickel, lead, and selenium in blood of the Brazillian population

Throughout the world, biomonitoring has become the standard for assessing exposure of individuals to toxic elements as well as for responding to serious environmental public health problems. However, extensive biomonitoring surveys require rapid and simple analytical methods. Thus, a simple and high-throughput method is proposed for the determination of arsenic (As), cadmium (Cd), copper (Cu), manganese (Mn), nickel (Ni), lead (Pb), and selenium (Se) in blood samples by using inductively coupled plasma–mass spectrometry (ICPMS). Prior to analysis, 200 ml of blood samples was mixed with 500 ml of 10% v/v tetramethylammonium hydroxide (TMAH) solution, incubated for 10 min, and subsequently diluted to 10 ml with a solution containing 0.05% w/v ethylenediamine tetraacetic acid (EDTA) + 0.005% v/v Triton X-100. After that, samples were directly analyzed by ICP-MS (ELAN DRC II). Rhodium was selected as an internal standard with matrix-matching calibration. Method detection limits were 0.08, 0.04, 0.5, 0.09, 0.12, 0.04, and 0.1 mg//L for As, Cd, Cu, Mn, Ni, Pb, and Se, respectively. Validation data are provided based on the analysis of blood samples from the trace elements inter-\comparison program operated by the Institut National de Santé Publique du Quebec, Canada. Additional validation was provided by the analysis of human blood samples by the proposed method and by using electrothermal atomic absorption spectrometry (ETAAS). The method was subsequently applied for the estimation of background metal blood values in the Brazilian population. In general, the mean concentrations of As, Cd, Cu, Mn, Ni, Pb, and Se in blood were 1.1, 0.4, 890, 9.6, 2.1, 65.4, and 89.3 mg/L, respectively, and are in agreement with other global populations. Influences of age, gender, smoking habits, alcohol consumption, and geographical variation on the values were also considered. Smoking habits influenced the levels of Cd in blood. The levels of Cu, Mn, and Pb were significantly correlated with gender, whereas Cu and Pb were significantly correlated with age. There were also interesting differences in Mn and Se levels in the population living in the north of Brazil compared to the south.

Variação bioacústica das vocalizações do complexo Taraba major (Vieillot, 1816) (Aves: Passeriformes: Thamnophilidae)

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

Biotic versus abiotic pollination in Oeceoclades maculata (Lindl.) Lindl. (Orchidaceae)

Geographical variation in the reproductive biology of widespread species often occurs at their distributional boundaries. We sought to determine whether such variation has occurred in an invasive orchid, Oeceoclades maculata, across its naturalized range. We compared its reproductive biology in a Brazilian population with that published for a population on the Caribbean island of Puerto Rico. In the state of Sao Paulo, O. maculata flowers between December and February, at the height of the rainy season. Similar fruit sets were observed in manual self (76%) and cross (70.4%) pollination treatments. The fruit set of plants protected from both pollinators and rainfall was 6.1%, whereas plants exposed only to rainfall had a fruit set of 41.4%, slightly less than the controls (48.3%). Like the Puerto Rico population, reproduction is primarily through rain-assisted autogamy, but unlike observations made on the island, outcrossing can eventually occur. We observed two butterfly species (Heliconius ethilla narcaea and Heliconius erato phyllis) pollinating O. maculata. Secretory epidermal cells and trichomes of the spur lumen produced 0.7 mu L of 25% (sucrose equivalents) nectar per flower each morning, which was stored in a dilated basal portion of the spur and reabsorbed by the afternoon. Thus, geographical variation in reproductive biology exists across the broad invasive range of O. maculata.