Evidence for Reductive Genome Evolution and Lateral Acquisition of Virulence Functions in Two Corynebacterium pseudotuberculosis Strains

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

The genome sequence of the gram-positive sugarcane pathogen Leifsonia xyli subsp. xyli

The genome sequence of Leifsonia xyli subsp. xyli, which causes ratoon stunting disease and affects sugarcane worldwide, was determined. The single circular chromosome of Leifsonia xyli subsp. xyli CTCB07 was 2.6 Mb in length with a GC content of 68% and 2,044 predicted open reading frames. The analysis also revealed 307 predicted pseudogenes, which is more than any bacterial plant pathogen sequenced to date. Many of these pseudogenes, if functional, would likely be involved in the degradation of plant heteropolysaccharides, uptake of free sugars, and synthesis of amino acids. Although L. xyli subsp. xyli has only been identified colonizing the xylem vessels of sugarcane, the numbers of predicted regulatory genes and sugar transporters are similar to those in free-living organisms. Some of the predicted pathogenicity genes appear to have been acquired by lateral transfer and include genes for cellulase, pectinase, wilt-inducing protein, lysozyme, and desaturase. The presence of the latter may contribute to stunting, since it is likely involved in the synthesis of abscisic acid, a hormone that arrests growth. Our findings are consistent with the nutritionally fastidious behavior exhibited by L. xyli subsp. xyli and suggest an ongoing adaptation to the restricted ecological niche it inhabits.

Taxonomic and phylogenetic relationships between neotropical species of ticks from genus Amblyomma (Acari: Ixodidae) inferred from second internal transcribed spacer sequences of rDNA

The accurate specific identification of ticks is essential for the study, control and prevention of tick-borne diseases. Herein, we determined ribosomal nucleotide sequences of the second internal transcribed spacer (ITS2) of 15 Neotropical hard tick species of the genus Amblyomma Koch found in Brazil. Most of the studied ticks accidentally parasite humans and potentially act as vectors of zoonoses. Lengths of the ITS2 sequences ranged from 956 to 1,207 bp, whereas GC content varied from 62.4 to 66.9%. A matrix of ITS2 divergence was calculated with the ITS2 sequence data obtained showing divergence levels varying from 1.5 to 28.8%. The analysis indicated that this molecular marker can be useful for Amblyomma-specific identification. Phylogenetic inferences based on the ITS2 sequences were used to assess some issues in subgenus taxonomy. © 2007 Entomological Society of America.

Characterization and genome organization of a repetitive element associated with the nucleolus organizer region in leporinuselongatus (anostomidae: Characiformes)

Chromosome mapping and studies of the genomic organization of repetitive DNA sequences provide valuable insights that enhance our evolutionary and structural understanding of these sequences, as well as identifying chromosomal rearrangements and sex determination. This study investigated the occurrence and organization of repetitive DNA sequences in Leporinus elongatus using restriction enzyme digestion and the mapping of sequences by chromosomal fluorescence in situ hybridization (FISH). A 378-bp fragment with a 54.2% GC content was isolated after digestion with the SmaI restriction enzyme. BLASTN search found no similarity with previously described sequences, so this repetitive sequence was named LeSmaI. FISH experiments were conducted using L. elongatus and other Anostomidae species, i.e. L. macrocephalus,L. obtusidens, L. striatus, L. lacustris, L. friderici, Schizodon borellii, S. isognathus, and Abramites hypselonotus which detected signals that were unique to male and female L. elongatus individuals. Double-FISH using LeSmaI and 18S rDNA showed that LeSmaI was located in a nucleolus organizer region (NOR) in the male and female metaphases of L. elongatus. This report also discusses the role of repetitive DNA associated with NORs in the diversification of Anostomidae species karyotypes. Copyright © 2012 S. Karger AG, Basel.

Triplet entropy analysis of hemagglutinin and neuraminidase sequences measures influenza virus phylodynamics

The influenza virus has been a challenge to science due to its ability to withstand new environmental conditions. Taking into account the development of virus sequence databases, computational approaches can be helpful to understand virus behavior over time. Furthermore, they can suggest new directions to deal with influenza. This work presents triplet entropy analysis as a potential phylodynamic tool to quantify nucleotide organization of viral sequences. The application of this measure to segments of hemagglutinin (HA) and neuraminidase (NA) of H1N1 and H3N2 virus subtypes has shown some variability effects along timeline, inferring about virus evolution. Sequences were divided by year and compared for virus subtype (H1N1 and H3N2). The nonparametric Mann-Whitney test was used for comparison between groups. Results show that differentiation in entropy precedes differentiation in GC content for both groups. Considering the HA fragment, both triplet entropy as well as GC concentration show intersection in 2009, year of the recent pandemic. Some conclusions about possible flu evolutionary lines were drawn. © 2013 Elsevier B.V.

Caracterização molecular e bioquímica de uma esterase macho-específica em Zaprionus indianus

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Ferramenta computacional para identificação de micro-organismos com base em assinaturas genômicas

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

A GC-FID Method to Determine Styrene in Polystyrene Glasses

The styrene levels of polystyrene (PS) glasses from the most consumed brands of disposable glasses intended for water and coffee in Brazil were determined. A GC-FID method was developed and validated, showing good precision and accuracy. The method was successfully used to determine styrene in 11 PS glass brands. The styrene levels ranged from 1.68 to 43.69 mg/100 g glass, depending on the kind of polymer, thickness, and glass brand. It could be used to control the content of styrene in the polymer. The migration of styrene from the glasses into water and 20 % ethanol was not detected.

Influence of different extraction methods on the yield and linalool content of the extracts of Eugenia uniflora L.

This work has been developed using a sylvestral fruit tree, native to the Brazilian forest, the Eugenia uniflora L., one of the Mirtaceae family. The main goal of the analytical study was focused on extraction methods themselves. The method development pointed to the Clevenger extraction as the best yield in relation to SFE and Soxhlet. The SFE method presented a good yield but showed a big amount of components in the final extract, demonstrating low selectivity. The essential oil extracted was analyzed by GC/FID showing a large range of polarity and boiling point compounds, where linalool, a widely used compound, was identified. Furthermore, an analytical solid phase extraction method was used to clean it up and obtain separated classes of compounds that were fractionated and studied by GC/FlD and GUMS. (c) 2006 Elsevier B.V. All rights reserved.

Influence of different drying temperatures on the amount of essential oils and citral content in Cymbopogon citratus (DC) Stapf. - Poaceae

Studies were conducted to show the effect of different temperatures in the drying process on the amount and quality of essential oils of Cymbopogon citratus (DC) Stapf. Leaves were harvested in the experimental field of the Agronomical Sciences College, UNESP, Botucatu, SP, Brazil in September, 1996. Blades of the leaves were cut in small parts (about 1-1,5 cm length), dried for several days at 30°, 50°, 70° and 90°C, until establishment of the weights. In the following process a hydrodistillation, during 2.5 hours, by Clevenger apparatus, was subsidized to extract the essential oils. A higher amount of oil could clearly be collected with the lower drying temperatures, except at 30°C, affected by fungus growing. Aspergillus sp., Penicillium sp., Rhyzopus sp., Cladosporium sp., Trichoderma sp. and Alternaria sp. were observed in the leaves. The analysis of the oil by GC-MS showed the variation of citral concentration of the treatments (86,1 to 95,2%). The results proved it is worthwhile to spend more time and effort in the production process using longer times of careful drying.

Drying temperature effects in peppermint essential oil content and composition

Studies were conducted to show the effect of different temperatures in the drying process on the amount and quality of essential oils of peppermint (Mentha piperita L.) The leaves were harvested in the Demeter Farmer, Botucatu, SP, Brazil in december, 1997. The leaves were dried at 40°C, 60°C and 80°C, until establishment of the weights. The essential oil was extracted by destilation in Clevenger apparatus and analysed by GC-MS. Higher drying temperature sharply decreased the essential oil content (% v/w) from 1.0% (40°C) to 0.14% (60°C) and 0.12% (80°C). Higher drying temperatures also affected the composition, decreasing the contents of 1,8 cineol and citronelal until 80°C, and increasing the contents of menthol and neomenthol until 60°C.

Effect of growth regulators in citral content in lemongrass in different seasons (in Portuguese)

The purpose of the present research was to evaluate the effect of plant growth regulators in yield citral content in lemongrass (Cymbopogon citratus (D.C.) Stapf), in different seasons. The field experiment was conducted on Sao Manuel Experimental Farm, Agronomical Science College, Paulista Estatal University, Botucatu, Brazil, which soil was classified in alic yellon red latosol (Haplorthox). Plants were randommly assigned blocks to treatments with three repetitions. The experimental groups consisted of GA3 (50 and 100 mg.l-1);Ethrel (100 and 200 mgl-1); CCC (500 and 1000 mg.l-1); Alar 85 (1000 and 2000 mg.l-1); Accel (20 and 40 mg.l-1) and control group. Five applications of plant growth regulators were realized every other three months. After 40 days of each foliar spray, the plants were cut and determined the citral yield. The essential oils were extracted from leaf tissue by hydrodistillation in a Clevenger apparatus. Samples of 250g of leaf and put in Clevenger apparatus with deionized water for 3h to determine the yield of essential oil. The citral content was analyzed by GC. In the present study the concentrations of plant growth regulators used did not increase citral content.

Determination of saturates, aromatics and ethanol in Brazilian commercial gasoline with low olefin content using 1 H NMR spectroscopy

In this work, a Hydrogen Nuclear Magnetic Resonance (1H NMR) method has been developed to determine aromatics and ethanol in Brazilian commercial gasoline with low olefin content. The proposed method involves subdividing an 1H NMR spectrum into regions, each of which is assumed to be associated with a specific type of structural group (OH, CH, CH2 and CH3). The method is based on the assignment of overlapping regions of 1H NMR spectra due to the signals of naphthene (N), iso and normal paraffins (P) and ethanol (E). Each 1H NMR spectrum was divided into 8 regions and the integration was correlated to the percentage of the substances to be determined. The results of the analysis by 1H NMR were compared with analysis of GC-FID obtained with the PONA system. The proposed technique of 1H NMR was shown to be an appropriate method for this sample type.