Edmar Vaz de Andrade

Guarana (Paullinia cupana var. sorbilis), an anciently consumed stimulant from the Amazon rain forest: the seeded-fruit transcriptome

Guarana (Paullinia cupana var. sorbilis) is a plant native to the central Amazon basin. Roasted seed extracts have been used as medicinal beverages since pre-Colombian times, due to their reputation as stimulants, aphrodisiacs, tonics, as well as protectors of the gastrointestinal tract. Guarana plants are commercially cultivated exclusively in Brazil to supply the national carbonated soft-drink industry and natural product stores around the world. In this report, we describe and discuss the annotation of 15,387 ESTs from guarana seeded-fruits, highlighting sequences from the flavonoid and purine alkaloid pathways, and those related to biotic stress avoidance. This is the largest set of sequences registered for the Sapindaceae family.

Mansonella ozzardi in Amazonas, Brazil: prevalence and distribution in the municipality of Coari, in the middle Solimões River

This study investigated some epidemiological aspects of the Mansonella ozzardi in municipality of Coari, Amazonas. Clinical symptoms were correlated with the filarial infection and the parasitic infection rates (PIR) were estimated in simuliid vectors. The general M. ozzardi human prevalence rate was 13.3% (231/1733), of which 10.2% (109/1069) were from the urban area and 18.4% (122/664) from the rural area. The prevalence rates were higher in men (14.5% urban and 19.7% rural) than in women (6.7% urban and 17.2% rural) and occurred in most age groups. The indices of microfilaremics were higher in people > or = 51 years old (26.9% urban and 61.5% rural). High prevalence rates were observed in retired people (27.1% urban area), housewives and farmer (41.6% and 25%, respectively, in rural area). The main clinical symptoms were joint pains and sensation of leg coldness. Only Cerqueirellum argentiscutum (Simuliidae) transmits M. ozzardi in this municipality (PIR = 5.6% urban and 7.1% rural). M. ozzardi is a widely distributed parasitic disease in Coari. Thus, temporary residency in the region of people from other localities involved with the local gas exploitation might be a contributing factor in spreading the disease.

Endo and exoglucanases produced by Penicillium citrinum isolated from Amazon

Background Cellulolytic enzymes (glucohidrolases EC 3.2.1.-) are biocatalizators highly specific. They act in synergy to hydrolyze b-1,4 bonds between monosaccharide units of D-glucose in the cellulose chain releasing its constituents. Cellulases are categorized according to the place they act in the cellulosic fiber. Endoglucanases start hydrolysis, exoglucanases act in the reduced terminal produced by endoglucanases followed by b-glycosidase which act in the product of exoglucanases catalysis releasing glucose monomers. Fungi are considered the best cellulolytic enzyme producers due to its natural cellulases that complete saccharification of lignocellulose. Species of Penicillium have been reported as excellent producers of cellulolytic enzymes when compared to commercial species and strains [1]. Aiming to contribute to biocatalytic processes and obtention of new sources for cellulolytic enzyme, this work has as objective the production of endoglucanases and exoglucanases from Penicillium citrinum isolated from an agro industrial residue in Amazon.

Sensitivity of diagnostic methods for Mansonella ozzardi microfilariae detection in the Brazilian Amazon Region.

BACKGROUND The human filarial worm Mansonella ozzardi is highly endemic in the large tributaries of the Amazon River. This infection is still highly neglected and can be falsely negative when microfilariae levels are low. OBJECTIVES This study investigated the frequency of individuals with M. ozzardi in riverine communities in Coari municipality, Brazilian Amazon. METHODS Different diagnostic methods including polymerase chain reaction (PCR), blood polycarbonate membrane filtration (PCMF), Knotts method (Knott), digital thick blood smears (DTBS) and venous thick blood smears (VTBS) were used to compare sensitivity and specificity among the methods. Data were analysed using PCMF and Bayesian latent class models (BLCM) as the gold standard. We used BLCM to calculate the prevalence of mansonelliasis based on the results of five diagnostic methods. FINDINGS The prevalence of mansonelliasis was 35.4% by PCMF and 30.1% by BLCM. PCR and Knott methods both possessed high sensitivity. Sensitivity relative to PCMF was 98.5% [95% confidence interval (CI): 92.0 - 99.7] for PCR and 83.5% (95% CI: 72.9 - 90.5) for Knott. Sensitivity derived by BLCM was 100% (95% CI 93.7 - 100) for PCMF, 100% (95% CI: 93.7 - 100) for PCR and 98.3% (95% CI: 90.6 - 99.9) for Knott. The odds ratio of being diagnosed as microfilaremic increased with age but did not differ between genders. Microfilariae loads were higher in subjects aged 30 - 45 and 45 - 60 years. MAIN CONCLUSIONS PCMF and PCR were the best methods to assess the prevalence of mansonelliasis in our samples. As such, using these methods could lead to higher prevalence of mansonelliasis in this region than the most commonly used method (i.e., thick blood smears).

Cloning andexpression of cDNA encoding growth hormone tambaqui (Colossoma macropomum) in the yeast Pichia pastoris

Genetic studies involving the search, cloning and expression of genes encoding proteins involved in important physiological processes and advantageous features of the economic standpoint have become the biotechnology research increasingly promising. Due to its zootechnical importance, the encoding gene of growth hormone (GH) of various fish species have been isolated, cloned and expressed in heterologous expression systems. The tambaqui (Colossoma macropomum), Amazonian fish species considered promising for fish farming, has been the subject of extensive genetic research with biotechnology focus. This study aimed to express the cDNA encoding GH tambaqui (tGH) in the methylotrophic yeast Pichia pastoris. The nucleotide sequence of cDNA tGH, previously isolated by Sousa (2009) [1], was optimized for expression in P. pastoris and obtained by chemical synthesis. It was then cloned into the cloning vector pUC19 followed by subcloning into expression and secretion vector pPIC9 with the endonucleases Eco RI and NotI. The recombinant plasmid named pPIC-tGH was linearized and inserted into host by electroporation. Recombinant clones were selected in medium auxotrophic for histidine. The expression of recombinant protein was induced by methanol addition during 96 hours occurred in shake flasks. The culture supernatant was analyzed on SDS-PAGE gel and expression of tGH was confirmed by Western blotting. The analysis of the supernatant revealed in both phenotype Mut+ and Muts the presence of a protein band of approximately 23 kDa, which was confirmed to be the tGH recombinant by Western blotting using monoclonal antibody against histidine tail C- terminal. The expression of tGH started already first 24 hours and was sustained throughout the induction period, lasted up to 120 h. These results are similar to of Li et al. [2], who expressed GH carp (Cyprinus carpio) in P. pastoris and observed that from the first 24 hours of induction the recombinant protein was already present in the medium, and time 72 and 96 hours corresponded to the peak of expression. These results demonstrate that GH tambaqui can be successfully expressed in P. pastoris. The tambaqui, for their economic importance, so far was the first Amazonian fish species to have GH expressed in heterologous system, the first step in the production of GHs of other important species for aquaculture regional and national. In the future, tambaqui recombinant GH should be analyzed for their efficiency in accelerating the growth of tambaqui towards the possible use in aquaculture production, which represent progress and innovation in technologies of cultivation of fish Amazonian.

Identification and proteomic analysis of petroleum degrading bacteria

Background The natural environment has suffered high accumulations of pollutants which affect not only the soil, groundwater, rivers, lakes, rainwater routes and air, but also human health due to the formation of unhealthy conditions. Excessive use of pesticides, crude oil spills, discharges of industrial waste, among other xenobiotic compounds accumulate in the environment and become worrisome for its persistence and its harmful effects to life. Therefore, bioremediation becomes an option for solving this problem, which is a tool that uses microorganisms to biochemical pollutants degradation compounds, transforming them into less or non-toxic substances. In this context, this work presents the results of identification of bacteria that degrade crude oil, as well as some results of a proteomic analysis of isolates grown in the presence of that pollutant to evaluate the profile of protein expression.

Purification of endoglucanase produced by Penicillium citrinum isolated from Amazon

Background The cellulolytic enzyme complex has many important biotechnological applications such as beverage, textile, food, paper and cellulose industries, as well as in degradation of lignocellulosic for ethanol biofuel production [1]. According to the enzymatic activity, cellulolytic complex is subdivided in three classes: endoglucanases, exoglucanases and b-D-glycosidases. Endoglucanases (endo-b-1,4-glucanase, EC 3.2.1.4) are responsible to initiate cleavage, hydrolyzing randomly internal regions from the amorphous cellulose fiber structures, releasing oligosaccharides from different grades of polymerization. These will be hydrolyzed by exoglucanases releasing cellobiose followed by b-D-glycosidades being hydrolyzed to glucose [1]. Penicillium citrinum has worldwide occurrence. Commonly in the soil, this specie is described as good xylanase and cellulase producer [2,3]. This work describes liquid chromatography to separate endoglucanases of Penicillium citrinum supernatant from submerse fermentation process.