Electrospray Ionization Mass Spectrometry Fingerprinting of Extracts of the Leaves of Arrabidaea chica

Arrabidaea chica (crajiru) is an important Amazonian plant. Its extracts are used as red pigments, antimicrobial agents and astringents. Three different varieties of this species are cultivated in the Amazon region. In this work, direct infusions of A. chica extracts from these three varieties were analyzed via electrospray ionization mass spectrometry (ESI(+)-MS) fingerprinting. Derived data from the spectra were classified by using a multivariate method (PLS-DA, partial least squares-discriminant analysis). The direct method that is herein presented relies on extraction of dry, powdered leaves with acidified methanol/water solution with no further sample preparation. The resulting supernatants were analyzed by direct infusion ESI(+)-MS, which provides characteristic fingerprints of the sample composition. 3-Deoxyanthocyanidins are important substances in A. chica, their ions were used as markers in the PLS-DA data treatment. PLS-DA was able to differentiate the three varieties. ESI(+)-MS fingerprinting works as a simple and fast method to differentiate varieties of A. chica.

A trophic study of the sympatric Amazonian freshwater turtles Podocnemis unifilis and Podocnemis expansa (Testudines, Podocnemidae) using carbon and nitrogen stable isotope analyses

The Yellow-spotted River Turtle (Podocnemis unifilis Troschel, 1848) and the South American River Turtle (Podocnemis expansa (Schweigger, 1812)) are two turtles species that are widely distributed and have ecological, economic, and cultural importance in the Amazon basin. Although sympatric regarding most of their distribution, few studies have addressed the coexistence of these two species. To examine this, we analyzed the trophic level and the primary carbon source from the diets of both species in Baixo Araguaia, Tocantins, Brazil, using stable isotope analyses of carbon (delta C-13) and nitrogen (delta N-15). We also verified possible intraspecific variations (related to sex and body mass) in the trophic levels and primary carbon sources of their diets. Podocnemis unifilis had higher values of delta N-15 than P. expansa, averaging 7.59 parts per thousand and 5.06 parts per thousand, respectively, a difference which may indicate a possible trophic change owing to exploiting different food resources. No differences were found between the two species in relation to delta C-13 (mean values of -26.2 parts per thousand and -26.1 parts per thousand, respectively). The similarity between delta C-13 values suggests that the sources of their basal feeding are the same, consisting mainly of C-3 plants. There was no intraspecific variation in the values of delta C-13 and delta N-15.

Natural infection of triatomines (Hemiptera: Reduviidae) by trypanosomatids in two different environments in the municipality of Ouro Preto do Oeste, State of Rondonia, Brazil

Introduction: This study analyzed the occurrence and the contamination of triatomines by trypanosomatids in Orbignya speciosa (babassu) specimens in the State of Rondonia, Brazil, in two different environments (pasture and woods). Methods: Capture of triatomines on babassus and microscopic search for trypanosomatids in their digestive tube were carried out. Results: Four hundred ninety-four (494) specimens were captured (Rhodnius prolixus and R.robustus), of which 35.6% of the triatomines were positive for trypanosomatids. Conclusions: The high index of natural infection along with the abundance of triatomines points out to the necessity to create an epidemiological surveillance system to monitor vector-borne transmission and deepen the studies on the ecology of such vectors in the Amazon.

Sedimentological and provenance response to Cambrian closure of the Clymene ocean: The upper Alto Paraguai Group, Paraguay belt, Brazil

Final Gondwana amalgamation was marked by the closure of the Neoproterozoic Clymene ocean between the Amazonia craton and central Gondwana. The events which occurred in the last stage of this closure were recorded in the upper Alto Paraguai Group in the foreland of the Paraguay orogen. Outcrop-based fades analysis of the siliciclastic rocks of upper Alto Paraguai Group, composed of the Sepotuba and Diamantino Formations, was carried out in the Diamantino region, within the eastern part of the Barra dos Bugres basin, Mato Grosso state, central-western Brazil. The Sepotuba Formation is composed of sandy shales with planar to wave lamination interbedded with fine-grained sandstone with climbing ripple cross-lamination, planar lamination, swaley cross-stratification and tangential to sigmoidal cross-bedding with mud drapes, related to marine offshore deposits. The lower Diamantino Formation is composed of a monotonous, laterally continuous for hundreds of metres, interbedded siltstone and fine-grained sandstone succession with regular parallel lamination, climbing ripple cross-lamination and ripple-bedding interpreted as distal turbidites. The upper part of this formation consists of fine to medium-grained sandstones with sigmoidal cross-bedding, planar lamination, climbing ripple cross-lamination, symmetrical to asymmetrical and linguoid ripple marks arranged in lobate sand bodies. These fades are interbedded with thick siltstone in coarsening upward large-scale cycles related to a delta system. The Sepotuba Formation characterises the last transgressive deposits of the Paraguay basin representing the final stage of a marine incursion of the Clymene ocean. The progression of orogenesis in the hinterland resulted in the confinement of the Sepotuba sea as a foredeep sub-basin against the edge of the Amazon craton. Turbidites were generated during the deepening of the basin. The successive filling of the basin was associated with progradation of deltaic lobes from the southeast, in a wide lake or a restricted sea that formed after 541 +/- 7 Ma. Southeastern to east dominant Neoproterozoic source regions were confirmed by zircon grains that yielded ages around 600 to 540 Ma, that are interpreted to be from granites in the Paraguay orogen. This overall regressive succession recorded in the Alto Paraguai Group represents the filling up of a foredeep basin after the final amalgamation of westem Gondwana in the earliest Phanerozoic. (C) 2011 International Association for Gondwana Research. Published by Elsevier B.V. All rights reserved.

EVALUATION OF GLUTATHIONE S-TRANSFERASE GSTM1 AND GSTT1 POLYMORPHISMS AND METHYLMERCURY METABOLISM IN AN EXPOSED AMAZON POPULATION

Over the last decades, the presence of methylmercury (MeHg) in the Amazon region of Brazil and its adverse human health effects have given rise to much concern. The biotransformation of MeHg occurs mainly through glutathione (GSH) in the bile mediated by conjugation with glutathione S-transferases (GST). Epidemiological evidence has shown that genetic polymorphisms may affect the metabolism of MeHg. The aim of this study was to evaluate the association between GST polymorphisms, GSH, and Hg levels in blood (B-Hg) and in hair (H-Hg) of an Amazon population chronically exposed to the metal through fish consumption. Blood and hair samples were collected from 144 volunteers (71 men, 73 women). B-Hg and H-Hg levels were determined by inductively coupled plasma-mass spectrometry, and GSH levels were evaluated by a spectrophotometric method. GSTM1 and T1 genotyping evaluation were carried out by multiplex polymerase chain reaction (PCR). Mean levels of B-Hg and H-Hg were 37.7 +/- 24.5 mu g/L and 10.4 +/- 7.4 mu g/g, respectively; GSH concentrations ranged from 0.52 to 2.89 mu M/ml of total blood. Distributions for GSTM1/T1, GSTM1/GSTT1*0, GSTM1*0/T1, and GSTM1*0/GSTT1*0 genotypes were 35.4, 22.2, 25.0, and 17.4%, respectively. GSTT1 genotype carriers presented lower levels of B-Hg and H-Hg when compared to other genotypes carriers. In addition, GSTM1*0/GSTT1*0 individuals presented higher Hg levels in blood and hair than subjects presenting any other genotypes. There appeared to be no evidence of an effect of polymorphisms on GSH levels. Therefore, our data suggest that GST polymorphisms may be associated with MeHg detoxification.

A New Species of Cascudinho of the Genus Hisonotus (Siluriformes: Loricariidae: Hypoptopomatinae) from the Upper Rio Tapajos Basin, Brazil

Hisonotus bockmanni, new species, is described based on specimens collected in a sandbank in the Rio Cururu, a tributary to the Rio Teles Pires, one of the rivers forming the Rio Tapajos in the Amazon Basin. The new taxon is distinguished from its congeners by a unique color pattern, whose most striking features are: two elliptical white spots, anterior to nostrils; predorsal region darkly pigmented with five unpigmented spots arranged as anteriorly pointed chevron; and a rostrocaudally elongate cross along most of the caudal peduncle. The placement of the new species in Hisonotus as well as its possible affinities within that genus are discussed in light of the current knowledge of the phylogenetic relationships among the Hypoptopomatinae.

Effect of artesunate-mefloquine fixed-dose combination in malaria transmission in amazon basin communities

Background: Studies in South-East Asia have suggested that early diagnosis and treatment with artesunate (AS) and mefloquine (MQ) combination therapy may reduce the transmission of Plasmodium falciparum malaria and the progression of MQ resistance. Methods: The effectiveness of a fixed-dose combination of AS and MQ (ASMQ) in reducing malaria transmission was tested in isolated communities of the Jurua valley in the Amazon region. Priority municipalities within the Brazilian Legal Amazon area were selected according to pre-specified criteria. Routine national malaria control programmatic procedures were followed. Existing health structures were reinforced and health care workers were trained to treat with ASMQ all confirmed falciparum malaria cases that match inclusion criteria. A local pharmacovigilance structure was implemented. Incidence of malaria and hospitalizations were recorded two years before, during, and after the fixed-dose ASMQ intervention. In total, between July 2006 and December 2008, 23,845 patients received ASMQ. Two statistical modelling approaches were applied to monthly time series of P. falciparum malaria incidence rates, P. falciparum/Plasmodium vivax infection ratio, and malaria hospital admissions rates. All the time series ranged from January 2004 to December 2008, whilst the intervention period span from July 2006 to December 2008. Results: The ASMQ intervention had a highly significant impact on the mean level of each time series, adjusted for trend and season, of 0.34 (95% CI 0.20 - 0.58) for the P. falciparum malaria incidence rates, 0.67 (95% CI 0.50 - 0.89) for the P. falciparum/P. vivax infection ratio, and 0.53 (95% CI 0.41 - 0.69) for the hospital admission rates. There was also a significant change in the seasonal (or monthly) pattern of the time series before and after intervention, with the elimination of the malaria seasonal peak in the rainy months of the years following the introduction of ASMQ. No serious adverse events relating to the use of fixed-dose ASMQ were reported. Conclusions: In the remote region of the Jurua valley, the early detection of malaria by health care workers and treatment with fixed-dose ASMQ was feasible and efficacious, and significantly reduced the incidence and morbidity of P. falciparum malaria.

The genetic diversity of Plasmodium malariae and Plasmodium brasilianum from human, simian and mosquito hosts in Brazil

Plasmodium malariae is a protozoan parasite that causes malaria in humans and is genetically indistinguishable from Plasmodium brasilianum, a parasite infecting New World monkeys in Central and South America. P. malariae has a wide and patchy global distribution in tropical and subtropical regions, being found in South America, Asia, and Africa. However, little is known regarding the genetics of these parasites and the similarity between them could be because until now there are only a very few genomic sequences available from simian Plasmodium species. This study presents the first molecular epidemiological data for P. malariae and P. brasilianum from Brazil obtained from different hosts and uses them to explore the genetic diversity in relation to geographical origin and hosts. By using microsatellite genotyping, we discovered that of the 14 human samples obtained from areas of the Atlantic forest, 5 different multilocus genotypes were recorded, while in a sample from an infected mosquito from the same region a different haplotype was found. We also analyzed the longitudinal change of circulating plasmodial genetic profile in two untreated non-symptomatic patients during a 12-months interval. The circulating genotypes in the two samples from the same patient presented nearly identical multilocus haplotypes (differing by a single locus). The more frequent haplotype persisted for almost 3 years in the human population. The allele Pm09-299 described previously as a genetic marker for South American P. malariae was not found in our samples. Of the 3 non-human primate samples from the Amazon Region, 3 different multilocus genotypes were recorded indicating a greater diversity among isolates of P. brasilianum compared to P. malariae and thus, P. malariae might in fact derive from P. brasilianum as has been proposed in recent studies. Taken together, our data show that based on the microsatellite data there is a relatively restricted polymorphism of P. malariae parasites as opposed to other geographic locations. (c) 2012 Elsevier B.V. All rights reserved.

Evaluation of the acquired immune responses to Plasmodium vivax VIR variant antigens in individuals living in malaria-endemic areas of Brazil

Abstract Background The naturally-acquired immune response to Plasmodium vivax variant antigens (VIR) was evaluated in individuals exposed to malaria and living in different endemic areas for malaria in the north of Brazil. Methods Seven recombinant proteins representing four vir subfamilies (A, B, C, and E) obtained from a single patient from the Amazon Region were expressed in Escherichia coli as soluble glutathione S-transferase fusion proteins. The different recombinant proteins were compared by ELISA with regard to the recognition by IgM, IgG, and IgG subclass of antibodies from 200 individuals with patent infection. Results The frequency of individuals that presented antibodies anti-VIR (IgM plus IgG) during the infection was 49%. The frequencies of individuals that presented IgM or IgG antibodies anti-VIR were 29.6% or 26.0%, respectively. The prevalence of IgG antibodies against recombinant VIR proteins was significantly lower than the prevalence of antibodies against the recombinant proteins representing two surface antigens of merozoites of P. vivax: AMA-1 and MSP119 (57.0% and 90.5%, respectively). The cellular immune response to VIR antigens was evaluated by in vitro proliferative assays in mononuclear cells of the individuals recently exposed to P. vivax. No significant proliferative response to these antigens was observed when comparing malaria-exposed to non-exposed individuals. Conclusion This study provides evidence that there is a low frequency of individuals responding to each VIR antigens in endemic areas of Brazil. This fact may explain the host susceptibility to new episodes of the disease.

Detection of DENV-4 genotype I from mosquitoes collected in the city of Manaus, Brazil

Background Dengue epidemics have been reported in Brazil since 1981. In Manaus, a large city in the Amazon region, dengue is endemic with all four-virus serotypes (DENV-1, -2, -3, and -4) simultaneously causing human disease. In 2008, during a surveillance of dengue virus in mosquitoes in the district of Tancredo Neves in Manaus, 260 mosquitoes of Aedes genus were captured, identified and grouped into pools of 10 mosquitoes. Findings RNA extracts of mosquito pools were tested by a RT-Hemi-Nested-PCR for detection of flaviviruses. One amplicon of 222 bp, compatible with dengue virus serotype 4, was obtained from a pool of Aedes aegypti. The nucleotide sequence of the amplicon indicated that the mosquitoes were infected with DENV-4 of genotype I. This virus of Asian origin has been described in Manaus in 2008 infecting acute febrile illness patients. Conclusion This is the first report of dengue virus serotype 4 genotype I infecting Aedes aegypti in the Americas.

Phylogeography of the neotropical Anopheles triannulatus complex (Diptera: Culicidae) supports deep structure and complex patterns

Abstract Background The molecular phylogenetic relationships and population structure of the species of the Anopheles triannulatus complex: Anopheles triannulatus s.s., Anopheles halophylus and the putative species Anopheles triannulatus C were investigated. Methods The mitochondrial COI gene, the nuclear white gene and rDNA ITS2 of samples that include the known geographic distribution of these taxa were analyzed. Phylogenetic analyses were performed using Bayesian inference, Maximum parsimony and Maximum likelihood approaches. Results Each data set analyzed septely yielded a different topology but none provided evidence for the seption of An. halophylus and An. triannulatus C, consistent with the hypothesis that the two are undergoing incipient speciation. The phylogenetic analyses of the white gene found three main clades, whereas the statistical parsimony network detected only a single metapopulation of Anopheles triannulatus s.l. Seven COI lineages were detected by phylogenetic and network analysis. In contrast, the network, but not the phylogenetic analyses, strongly supported three ITS2 groups. Combined data analyses provided the best resolution of the trees, with two major clades, Amazonian (clade I) and trans-Andean + Amazon Delta (clade II). Clade I consists of multiple subclades: An. halophylus + An. triannulatus C; trans-Andean Venezuela; central Amazonia + central Bolivia; Atlantic coastal lowland; and Amazon delta. Clade II includes three subclades: Panama; cis-Andean Colombia; and cis-Venezuela. The Amazon delta specimens are in both clades, likely indicating local sympatry. Spatial and molecular variance analyses detected nine groups, corroborating some of subclades obtained in the combined data analysis. Conclusion Combination of the three molecular markers provided the best resolution for differentiation within An. triannulatus s.s. and An. halophylus and C. The latest two species seem to be very closely related and the analyses performed were not conclusive regarding species differentiation. Further studies including new molecular markers would be desirable to solve this species status question. Besides, results of the study indicate a trans-Andean origin for An. triannulatus s.l. The potential implications for malaria epidemiology remain to be investigated.

Natural infection of triatomines (Hemiptera: Reduviidae) by trypanosomatids in two different environments in the municipality of Ouro Preto do Oeste, State of Rondônia, Brazil

INTRODUCTION: This study analyzed the occurrence and the contamination of triatomines by trypanosomatids in Orbignya speciosa (babassu) specimens in the State of Rondônia, Brazil, in two different environments (pasture and woods). METHODS: Capture of triatomines on babassus and microscopic search for trypanosomatids in their digestive tube were carried out. RESULTS: Four hundred ninety-four (494) specimens were captured (Rhodnius prolixus and R.robustus), of which 35.6% of the triatomines were positive for trypanosomatids. CONCLUSIONS: The high index of natural infection along with the abundance of triatomines points out to the necessity to create an epidemiological surveillance system to monitor vector-borne transmission and deepen the studies on the ecology of such vectors in the Amazon.

Landscape and soil regionalization in southern Brazilian Amazon and contiguous areas: methodology and relevance for ecological studies

Soils of a large tropical area with differentiated landscapes cannot be treated uniformly for ecological applications. We intend to develop a framework based on physiography that can be used in regional applications. The study region occupies more than 1.1 million km² and is located at the junction of the savanna region of Central Brazil and the Amazon forest. It includes a portion of the high sedimentary Central Brazil plateau and large areas of mostly peneplained crystalline shield on the border of the wide inner-Amazon low sedimentary plain. A first broad subdivision was made into landscape regions followed by a more detailed subdivision into soil regions. Mapping information was extracted from soil survey maps at scales of 1:250000-1:500000. Soil units were integrated within a homogenized legend using a set of selected attributes such as taxonomic term, the texture of the B horizon and the associated vegetation. For each region, a detailed inventory of the soil units with their area distribution was elaborated. Ten landscape regions and twenty-four soil regions were recognized and delineated. Soil cover of a region is normally characterized by a cluster composed of many soil units. Soil diversity is comparable in the landscape and the soil regions. Composition of the soil cover is quantitatively expressed in terms of area extension of the soil units. Such geographic divisions characterized by grouping soil units and their spatial estimates must be used for regional ecological applications.

Three new species of Lebiasina (Characiformes: Lebiasinidae) from the Brazilian shield border at Serra do Cachimbo, Pará, Brazil

Lebiasina marilynae n. sp., L. melanoguttata n. sp., and L. minuta n. sp. are described from the headwaters of the rio Curuá, in Serra do Cachimbo, Pará, Brazil, and represent the only members of the Lebiasininae in the Brazilian Shied, so far. A close relationship among these species is proposed based on: 1) the presence of a pair of foramina through which the ramus palatinus of the facial nerve passes, a modification unique in Lebiasinidae and apparently in the Characiformes, 2) the enlargement of the extrascapular bone, 3) the absence of the secondary stripe, and 4) the nearly equal length of caudal-fin lobes. Lebiasina marilynae additionally differs from all congeners in having the primary stripe extending from the tip of the snout to the distal border of the caudal-fin peduncle, the possession of two series of dark blotches parallel to the primary stripe, and a rounded dorsal surface of the mesethmoid. Lebiasina melanoguttata and Lebiasina minuta additionally differ from all congeners in the absence of the primary stripe and the caudal blotch, and the presence of three longitudinal series of dark blotches at the base of the scales of series 3-5. Lebiasina melanoguttata differs from Lebiasina minuta in the absence of a dark blotch at the base of the median rays of the dorsal fin, second infrapharyngobranchial bearing conical teeth, the reddish overall coloration of the eye and fins, and the dark blotches never coalescing (vs. dark dorsal-fin blotch present; the second infrapharyngobranchial being edentulous; dark, olive green eyes, and the yellowish overall color of body and fins; and the dark blotches of longitudinal series 3 and 4 coalescing where scales of adjacent longitudinal series overlap). The occurrence of species of the Lebiasininae on the Brazilian Shield is discussed, and the distribution pattern of the species described herein is compared to that of other endemic species of the Serra do Cachimbo, a highly biodiverse area isolated from the rest of the Amazon basin.

The dynamics of a frictionally-dominated Amazonian estuary

The hydrodynamics, morphology and sedimentology of the Taperaçu estuary were investigated. This is one of several estuaries located within the largest mangrove fringe in the world, bordering the Amazon region, subject to a macrotidal regime and regionally atypical negligible fresh water supply. The results reveal widespread sand banks that occupy the central portion of the estuarine cross-section. Well-sorted very fine sandy sediments of marine origin prevail. Shorter flood phases, with substantially higher current velocities, were observed in the upper sector of Taperaçu, as expected for a shallow, friction-dominated estuary. However, ebb domination can be expected for estuaries with large associated mangrove areas and substantial estuarine infilling, both of which situations occur on the Taperaçu. The tidal asymmetry favoring flood currents could be the result of the absence of an effective fluvial discharge. Furthermore, it was observed that the Taperaçu is connected by tidal creeks to the neighboring Caeté estuary, allowing a stronger flux during the flood and intensifying the higher flood currents. As a whole, the results have shown a complex interaction of morphological aspects (friction, fluvial drainage, connections with neighbor estuaries, infilling and large storage area) in determining hydrodynamic patterns, thus improving the understanding of Amazon estuaries.