Partial molecular characterization of the Nile tilapia (Oreochromis niloticus) alpha-cardiac muscle actin gene and its relationship to actin isoforms of other fish species

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

SEX-CHROMOSOME POLYMORPHISM AND HETEROGAMETIC MALES REVEALED BY 2 CLONED DNA PROBES IN THE ZW ZZ FISH LEPORINUS-ELONGATUS

In order to study the divergence of teleost sex chromosomes, subtractive cloning was carried out between genomic DNA of males and females of the rainbow trout (XX/XY) and of Leporinus elongatus (ZW/ZZ). Inserts cloned in a plasmid vector were individually tested on Southern blots of DNA of males and females for sex specificity. No sex-specific insert was obtained from trout, but two out of ten inserts cloned from L. elongatus showed sex-specific patterns in this species: one corresponds to a sequence present on both Z and W chromosomes, while the other is W specific. Sequences of these two inserts show neither clear homology with other known sequences, nor an open reading frame. They cross-hybridize with the genomic DNA of Leporinus friderici, but without sex-specific patterns. Twenty-four L. elongatus adults were sexed by gonadal observation, chromosomed examination and Southern hybridization with one or the other insert. Ten males and 11 females had chromosomes and hybridization patterns typical of their sex. One ZW female was recognized as a male with the W-specific probe. This was also the case for two unusual ZW males, one having a male hybridization pattern with the other probe. These three atypical individuals may result from single genetic exchanges between four regions of the Z and the W, giving rise to three atypical W chromosomes. Finding males with such atypical heterochromosomes in a female heterogametic species may indicate that a gradual transition occurs between the heterogametic systems.

The copia retrotransposon and horizontal transfer in Drosophila willistoni

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

Northwestern overthrusting and related lateral escape during the Brasiliano orogeny north of the Patos lineament, Borborema province, northeast Brazil

The Borborema province (BP) of northeastern Brazil, located between the São Luís and São Francisco cratons, represents a branching system of Precambrian orogens of the South American platform. It is composed of segments of Archean and Proterozoic crust that were deformed by the convergence of the West African and São Francisco-Congo cratons during assembly of the Brasiliano collage (650 to 500 Ma), a period of intense orogenic activity considered to be the strongest and most pervasive tectonic event that affected the Precambrian of the South American platform. The tectonic and kinematic history of the Brasiliano/Panafrican orogeny is fundamental for reconstructing South American and African Precambrian geology. The correlation between Neoproterozoic tectonic processes occurring in both continents should use structural elements, of regional or local character, with identical kinematic and metamorphic conditions manifested in both basement and supracrustal units. North of the Patos shear zone, subhorizontal Brasiliano thrusts (0.65 to 0.58 Ga) affected the basement and the supracrustal Seridó belt with such related regional D1/D2 structures as foliation, lineation, isoclinal folds, and related metamorphism. Overprinting the previous structures, regional folding with a vertical S3 foliation and an associated strike-slip shear zone were developed (0.58 to 0.52 Ga). The metamorphism is similar for all deformation phases, ranging from upper-greenschist to amphibolite facies with mineral assemblages including biolite and garnet throughout the Seridó fold belt. We propose, on the basis of deformational and kinematic reconstructions, that the structural evolution of the Seridó fold belt was characterized by transition from a syn-collisional to a strike-slip regime. The transition between regimes occurred, progressively or instantaneously, by the switching of the maximum and intermediary strain axes of the strain ellipsoid. The entire tectonic history can be related to a frontal or oblique collision and lateral escape tectonics, with local, syn-collisional transpression and transtension. The Patos shear zone represents a final vertical shearing, juxtaposing different terranes of the northern and southern Borborema province.

Strong selection against hybrids at a hybrid zone in the Ensatina ring species complex and its evolutionary implications

The analysis of interactions between lineages at varying levels of genetic divergence can provide insights into the process of speciation through the accumulation of incompatible mutations. Ring species, and especially the Ensatina eschscholtzii system exemplify this approach. The plethodontid salamanders E. eschscholtzii xanthoptica and E. eschscholtzii platensis hybridize in the central Sierran foothills of California. We compared the genetic structure across two transects (southern and northern Calaveras Co.), one of which was resampled over 20 years, and examined diagnostic molecular markers (eight allozyme loci and mitochondrial DNA) and a diagnostic quantitative trait (color pattern). Key results across all studies were: (1) cline centers for all markers were coincident and the zones were narrow, with width estimates of 730 m to 2000 m; (2) cline centers at the northern Calaveras transect were coincident between 1981 and 2001, demonstrating repeatability over five generations; (3) there were very few if any putative F1s, but a relatively high number of backcrossed individuals in the central portion of transects: and (4) we found substantial linkage disequilibrium in all three studies and strong heterozygote deficit both in northern Calaveras, in 2001, and southern Calaveras. Both linkage disequilibrium and heterozygote deficit showed maximum values near the center of the zones. Using estimates of cline width and dispersal, we infer strong selection against hybrids. This is sufficient to promote accumulation of differences at loci that are neutral or under divergent selection, but would still allow for introgression of adaptive alleles. The evidence for strong but incomplete isolation across this centrally located contact is consistent with theory suggesting a gradual increase in postzygotic incompatibility between allopatric populations subject to divergent selection and reinforces the value of Ensatina as a system for the study of divergence and speciation at multiple stages. © 2005 The Society for the Study of Evolution. All rights reserved.

A comparison of mycolic acid analysis for nontuberculous mycobacteria identification by thin-layer chromatography and molecular methods

The development of fast, inexpensive, and reliable tests to identify nontuberculous mycobacteria (NTM) is needed. Studies have indicated that the conventional identification procedures, including biochemical assays, are imprecise. This study evaluated a proposed alternative identification method in which 83 NTM isolates, previously identified by conventional biochemical testing and in-house M. avium IS1245-PCR amplification, were submitted to the following tests: thin-layer chromatography (TLC) of mycolic acids and PCR-restriction enzyme analysis of hsp65 (PRA). High-performance liquid chromatography (HPLC) analysis of mycolic acids and Southern blot analysis for M. avium IS1245 were performed on the strains that evidenced discrepancies on either of the above tests. Sixty-eight out of 83 (82%) isolates were concordantly identified by the presence of IS1245 and PRA and by TLC mycolic acid analysis. Discrepant results were found between the phenotypic and molecular tests in 12/83 (14.4%) isolates. Most of these strains were isolated from non-sterile body sites and were most probably colonizing in the host tissue. While TLC patterns suggested the presence of polymycobacterial infection in 3/83 (3.6%) cultures, this was the case in only one HPLC-tested culture and in none of those tested by PRA. The results of this study indicated that, as a phenotypic identification procedure, TLC mycolic acid determination could be considered a relatively simple and cost-effective method for routine screening of NTM isolates in mycobacteriology laboratory practice with a potential for use in developing countries. Further positive evidence was that this method demonstrated general agreement on MAC and M. simiae identification, including in the mixed cultures that predominated in the isolates of the disseminated infections in the AIDS patients under study. In view of the fact that the same treatment regimen is recommended for infections caused by these two species, TLC mycolic acid analysis may be a useful identification tool wherever molecular methods are unaffordable.

Visual and acoustic signaling in three species of Brazilian nocturnal tree frogs (anura, hylidae)

Visual communication seems to be widespread among nocturnal anurans, however, reports of these behaviors in many Neotropical species are lacking. Therefore, we gathered information collected during several sporadic field expeditions in central and southern Brazil with three nocturnal tree frogs: Aplastodiscus perviridis, Hypsiboas albopunctatus and H. bischoffi. These species displayed various aggressive behaviors, both visual and acoustic, towards other males. For A. perviridis we described arm lifting and leg kicking; for H. albopunctatus we described the advertisement and territorial calls, visual signalizations, including a previously unreported behavior (short leg kicking), and male-male combat; and for H. bischoffi we described the advertisement and fighting calls, toes and fingers trembling, leg lifting, and leg kicking. We speculate about the evolution of some behaviors and concluded that the use of visual signals among Neotropical anurans may be much more common than suggested by the current knowledge. © 2007 Departamento de Ciências Biológicas.

Determination of calcium and magnesium in hydroethanolic extracts of propolis by atomic absorption flame spectrophotometry

Propolis is a natural product collected by honeybees and has a large range of pharmacological activity, including antimicrobial, antitumoral, antioxidant and anti-inflammatory. Its use as a popular medicine is increasing all over the world, creating a need for quality control of the commercial products. In this study the levels of calcium and magnesium in commercial hydroalcoholic propolis extracts from varies states of Brazil were determined by atomic absorption flame spectrophotometry and different values were obtained for northern and southern states. This study can be extended to the analysis of metals that are harmful to health. The results showed that the calibration curves were linear over a wide concentration range (0.5-4.0 μg.mL -1 for calcium and 0.05-0.4 μg.mL -1 for magnesium) with good correlation coefficients (0.999 and 0.988, respectively). Good analytical recovery (94%) was obtained. The proposed method showed adequate precision and relative standard deviation lower than 2 %. The method is accurate and precise as well as having advantages such as simplicity and speed.

Molecular phylogenetic relationships and phenotypic diversity in miniaturized toadlets, genus Brachycephalus (Amphibia: Anura: Brachycephalidae)

Toadlets of the genus Brachycephalus are endemic to the Atlantic rainforests of southeastern and southern Brazil. The 14 species currently described have snout-vent lengths less than 18. mm and are thought to have evolved through miniaturization: an evolutionary process leading to an extremely small adult body size. Here, we present the first comprehensive phylogenetic analysis for Brachycephalus, using a multilocus approach based on two nuclear (Rag-1 and Tyr) and three mitochondrial (Cyt b, 12S, and 16S rRNA) gene regions. Phylogenetic relationships were inferred using a partitioned Bayesian analysis of concatenated sequences and the hierarchical Bayesian method (BEST) that estimates species trees based on the multispecies coalescent model. Individual gene trees showed conflict and also varied in resolution. With the exception of the mitochondrial gene tree, no gene tree was completely resolved. The concatenated gene tree was completely resolved and is identical in topology and degree of statistical support to the individual mtDNA gene tree. On the other hand, the BEST species tree showed reduced significant node support relative to the concatenate tree and recovered a basal trichotomy, although some bipartitions were significantly supported at the tips of the species tree. Comparison of the log likelihoods for the concatenated and BEST trees suggests that the method implemented in BEST explains the multilocus data for Brachycephalus better than the Bayesian analysis of concatenated data. Landmark-based geometric morphometrics revealed marked variation in cranial shape between the species of Brachycephalus. In addition, a statistically significant association was demonstrated between variation in cranial shape and genetic distances estimated from the mtDNA and nuclear loci. Notably, B. ephippium and B. garbeana that are predicted to be sister-species in the individual and concatenated gene trees and the BEST species tree share an evolutionary novelty, the hyperossified dorsal plate. © 2011 Elsevier Inc.

Morphological description of the nymphal stage of Amblyomma geayi and new nymphal records of Amblyomma parkeri

The external morphology of the nymph of Amblyomma geayi Neumann is described by optical and scanning electron microscopy. Unfed nymphs were obtained from an engorged A. geayi female, which had been collected on a sloth (Bradypus variegatus) from Belém municipality, State of Pará, northern Brazil, and was kept under laboratory conditions. With the present description, we propose a modification of a taxonomic key published in 2010 for the Amblyomma nymphs that occur in Brazil, through the inclusion of A. geayi. The nymph of A. geayi is morphologically very similar to the nymph of Amblyomma parkeri Fonseca and Aragão, with only slight morphological differences related to scutal surface and punctuations (more shagreened and less punctuated in A. geayi). These 2 nymphs differ from all other known Amblyomma nymphs from Brazil by the combination of auriculae present as small posterolateral rounded projections, eyes located at the level of the scutal midlength, and a rounded hypostome. These nymphal similarities as well the morphology of the adult stage corroborate previous studies that showed that A. geayi and A. parkeri are genetically closely related. Unpublished host records of the nymphs of both A. geayi and A. parkeri are provided. Established populations of A. geayi and A. parkeri seem to be geographically separated, since all confirmed records of A. geayi are from the northern half of South America (mainly the Amazonian region) and Central America, whereas all known records of A. parkeri are from the Atlantic rainforest biome in northeastern, southeastern, and southern Brazil. © 2013 Elsevier GmbH.

Revisiting AFLP fingerprinting for an unbiased assessment of genetic structure and differentiation of taurine and zebu cattle

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

From west to east: Environmental influences on the rate and pathways of Polynesian colonization

The prime movers behind the prehistoric colonization of Remote Oceania, and in particular the large c. 2000-year temporal gap (i.e. long pause') seen between West and East Polynesia, has long been major point of interest in the Pacific. To address these events and the processes that may have led to the known chronological disparity of these diasporas, we present results from two different, but equally powerful, analytical tools which are used to examine Polynesian seafaring capabilities and trajectories. The first is a statistical model known as Seascape, which simulates voyages, while the second uses ease of eastward travel estimates based on land distribution and wind pattern analysis. These analyses were done with the goal of determining the potential role of environmental factors in the colonization process, particularly as they relate to the long pause. We show that the eastern boundary of West Polynesia, the limit of the initial colonization pulse, is marked by a discontinuity in land distribution, where the distances travelers would have to cross in order to reach islands further to the east become significantly larger. At the same time, in West Polynesia, the frequency and intensity of winds favorable to eastward displacement decrease continuously from west to east. As far as winds are concerned, eastward travel in West Polynesia is favored in the northern and southern areas and much more difficult across the central portion. Favorable winds have a clear seasonality, and eastward displacement along the northern area is much easier under El Nino conditions. Voyaging simulations show that intentional eastward voyages departing from Tonga and Samoa, when undertaken with vessels capable of sailing efficiently against the wind, afford a viable route toward several island groups in East Polynesia, with trips starting in Samoa having a higher probability of success.

Population Structure and Pathotype Diversity of the Wheat Blast Pathogen Magnaporthe oryzae 25 Years After Its Emergence in Brazil

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

Trypanosoma cruzi strains from triatomine collected in Bahia and Rio Grande do Sul, Brazil

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Cytotoxic and genotoxic effects of ethanolic extract of Euphorbia hyssoptfolia L. on HepG2 cells

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)