Construction and characterisation of a complete reverse genetics system of dengue virus type 3

Dengue virulence and fitness are important factors that determine disease outcome. However, dengue virus (DENV) molecular biology and pathogenesis are not completely elucidated. New insights on those mechanisms have been facilitated by the development of reverse genetic systems in the past decades. Unfortunately, instability of flavivirus genomes cloned in Escherichia coli has been a major problem in these systems. Here, we describe the development of a complete reverse genetics system, based on the construction of an infectious clone and replicon for a low passage DENV-3 genotype III of a clinical isolate. Both constructs were assembled into a newly designed yeast- E. coli shuttle vector by homologous recombination technique and propagated in yeast to prevent any possible genome instability in E. coli . RNA transcripts derived from the infectious clone are infectious upon transfection into BHK-21 cells even after repeated passages of the plasmid in yeast. Transcript-derived DENV-3 exhibited growth kinetics, focus formation size comparable to original DENV-3 in mosquito C6/36 cell culture. In vitro characterisation of DENV-3 replicon confirmed its identity and ability to replicate transiently in BHK-21 cells. The reverse genetics system reported here is a valuable tool that will facilitate further molecular studies in DENV replication, virus attenuation and pathogenesis.

Spatial and temporal country-wide survey of temephos resistance in Brazilian populations of Aedes aegypti

The organophosphate temephos has been the main insecticide used against larvae of the dengue and yellow fever mosquito (Aedes aegypti) in Brazil since the mid-1980s. Reports of resistance date back to 1995; however, no systematic reports of widespread temephos resistance have occurred to date. As resistance investigation is paramount for strategic decision-making by health officials, our objective here was to investigate the spatial and temporal spread of temephos resistance in Ae. aegypti in Brazil for the last 12 years using discriminating temephos concentrations and the bioassay protocols of the World Health Organization. The mortality results obtained were subjected to spatial analysis for distance interpolation using semi-variance models to generate maps that depict the spread of temephos resistance in Brazil since 1999. The problem has been expanding. Since 2002-2003, approximately half the country has exhibited mosquito populations resistant to temephos. The frequency of temephos resistance and, likely, control failures, which start when the insecticide mortality level drops below 80%, has increased even further since 2004. Few parts of Brazil are able to achieve the target 80% efficacy threshold by 2010/2011, resulting in a significant risk of control failure by temephos in most of the country. The widespread resistance to temephos in Brazilian Ae. aegypti populations greatly compromise effective mosquito control efforts using this insecticide and indicates the urgent need to identify alternative insecticides aided by the preventive elimination of potential mosquito breeding sites.

Identification of care needs of patients with and without the use of a classification instrument

Objective: To analyze the agreement and disagreement between the assessments by applying or not a patient classification instrument, and to investigate the association between the agreement and personal and professional characteristics of the evaluators. Method: This is a descriptive exploratory study. 105 patients were hospitalized in a teaching hospital in the state of Sao Paulo, using the kappa statistic (weighted) and the Bootstrap method. Results: The agreement between the assessments were​​: kw 0.87 (instrument x internal evaluator), kw 0.78 (instrument x external evaluator) and kw 0.76 (between evaluators) and the influence of some personal and professional characteristics. The assessments conducted through the use of an instrument contemplated a greater number of areas of care in relation to when the instrument was not applied. Conclusion: The use of this instrument is recommended in order to more effectively identify care needs of patients.




Terms of international classification for Nursing Practice in motor and physical rehabilitation

OBJECTIVE To validate terms of nursing language especially for physical-motor rehabilitation and map them to the terms of ICNP® 2.0. METHOD A methodology research based on document analysis, with collection and analysis of terms from 1,425 records. RESULTS 825 terms were obtained after the methodological procedure, of which 226 had still not been included in the ICNP® 2.0. These terms were distributed as follows: 47 on the Focus axis; 15 on the Judgment axis; 31 on the Action axis; 25 on the Location axis; 102 on the Means axis; three on the Time axis; and three on the Client axis. All non-constant terms in ICNP® have been validated by experts, having reached an agreement index ≥0.80. CONCLUSION The ICNP® is applicable and used in nursing care for physical-motor rehabilitation.

New Brazilian species of Asphondyliini (Diptera, Cecidomyiidae)

Asphondylia canastrae sp. nov. (Minas Gerais, Delfinópolis), A. sanctipetri sp. nov. (São Paulo, Ribeirão Preto), and Schizomyia tuiuiu sp. nov. (Mato Grosso, Cuiabá) are described. A. canastrae sp. nov. is associated with Hyptis sp. (Lamiaceae), A. sanctipetri sp. nov. with Didymopanax morototoni (Araliaceae), and S. tuiuiu sp. nov. with Bauhinia rufa (Fabaceae). Illustrations of the new species and comments about their systematic position are presented.

Saving our science from ourselves: the plight of biological classification

Saving our science from ourselves: the plight of biological classification. Biological classification ( nomenclature, taxonomy, and systematics) is being sold short. The desire for new technologies, faster and cheaper taxonomic descriptions, identifications, and revisions is symptomatic of a lack of appreciation and understanding of classification. The problem of gadget-driven science, a lack of best practice and the inability to accept classification as a descriptive and empirical science are discussed. The worst cases scenario is a future in which classifications are purely artificial and uninformative.

A pictorial key and diagnosis of the Brazilian genera of Micropezidae (Diptera, Nerioidea)

A pictorial key and diagnosis of the Brazilian genera of Micropezidae (Diptera, Nerioidea). This paper provides the first pictorial key and diagnosis for the Brazilian genera of the Micropezidae, based on external morphological characters illustrated with photographs. The key includes 13 genera: Cardiacephala Macquart, Cliobata Enderlein, Grallipeza Rondani, Metasphen Frey, Micropeza Meigen, Parasphen Enderlein, Planipeza Marshall, Plocoscelus Enderlein, Poecilotylus Hennig, Ptilosphen Enderlein, Rainieria Rondani, Scipopus Enderlein and Taeniaptera Macquart. For each genus, the species known to occur in Brazil are listed and their distribution records, including new ones, are provided.

Assessment of soil properties by organic matter and EM-microorganism incorporation

Properties of a claim loam soil, collected in Aranjuez (Madrid) and enriched with organic matter and microorganisms, were evaluated under controlled temperature and moisture conditions, over a period of three months. The following treatments were carried out: soil (control); soil + 50 t ha-1 of animal manure (E50); soil + 50 t ha-1 of animal manure + 30 L ha-1 of effective microorganisms (E50EM); soil + 30 t ha-1 of the combination of various green crop residues and weeds (RC30) and soil + 30 t ha-1 of the combination of various green crop residues and weeds + 30 L ha-1 of effective microorganisms (RC30EM). Soil samples were taken before and after incubation and their physical, chemical, and microbiological parameters analyzed. Significant increase was observed in the production of exopolysaccharides and basic phosphatase and esterase enzyme activities in the treatments E50EM and RC30EM, in correlation with the humification of organic matter, water retention at field capacity, and the cationic exchange capacity (CEC) of the same treatments. The conclusion was drawn that the incorporation of a mixture of effective microorganisms (EM) intensified the biological soil activity and improved physical and chemical soil properties, contributing to a quick humification of fresh organic matter. These findings were illustrated by the microbiological activities of exopolysaccharides and by alkaline phosphatase and esterase enzymes, which can be used as early and integrated soil health indicators.

Comparison of estimation methods of soil strength in five soils

In agriculture, the soil strength is used to describe the susceptibility to deformation by pressure caused by agricultural machine. The purpose of this study was to compare different methods for estimating the inherent soil strength and to identify their suitability for the evaluation of load support capacity, compaction susceptibility and root growth. The physical, chemical, mineralogical and intrinsic strength properties of seven soil samples, collected from five sampling pits at different locations in Brazil, were measured. Four clay (CS) and three sandy clay loam (SCL) soils were used. The clay soils were collected on a farm in Santo Ângelo, RS (28 º 16 ' 16 '' S; 54 º 13 ' 11 '' W 290 m); A and B horizons at the Universidade Federal de Lavras, Lavras, MG (21 º 13 ' 47 '' S; 44 º 58 ' 6'' W; 918 m) and on the farm Sygenta, in Uberlandia, MG (18 º 58 ' 37 '' S; 48 º 12 ' 05 '' W 866 m). The sandy clay loam soils were collected in Aracruz, ES (19 º 47 ' 10 '' S; 40 º 16 ' 29 '' W 81 m), and on the farm Xavier, Lavras, MG (21 º 13 ' 24 '' S; 45 º 05 ' 00 '' W; 844 m). Soil strength was estimated based on measurements of: (a) a pneumatic consolidometer, (b) manual pocket (non-rotating) penetrometer; and (c) automatic (rotating) penetrometer. The results of soil strength properties were similar by the three methods. The soil structure had a significant influence on soil strength. Results of measurements with both the manual pocket and the electric penetrometer were similar, emphasizing the influence of soil texture. The data showed that, to enhance the reliability of predictions of preconsolidation pressure by penetrometers, it is better to separate the soils into the different classes, rather than analyze them jointly. It can be concluded that the consolidometer method, although expensive, is the best when evaluations of load support capacity and compaction susceptibility of soil samples are desired.

Spatial variability of soil chemical properties after coffee tree removal

Assessing the spatial variability of soil chemical properties has become an important aspect of soil management strategies with a view to higher crop yields with minimal environmental degradation. This study was carried out at the Centro Experimental of the Instituto Agronomico, in Campinas, São Paulo, Brazil. The aim was to characterize the spatial variability of chemical properties of a Rhodic Hapludox on a recently bulldozer-cleaned area after over 30 years of coffee cultivation. Soil samples were collected in a 20 x 20 m grid with 36 sampling points across a 1 ha area in the layers 0.0-0.2 and 0.2-0.4 m to measure the following chemical properties: pH, organic matter, K+, P, Ca2+, Mg2+, potential acidity, NH4-N, and NO3-N. Descriptive statistics were applied to assess the central tendency and dispersion moments. Geostatistical methods were applied to evaluate and to model the spatial variability of variables by calculating semivariograms and kriging interpolation. Spatial dependence patterns defined by spherical model adjusted semivariograms were made for all cited soil properties. Moderate to strong degrees of spatial dependence were found between 31 and 60 m. It was still possible to map soil spatial variability properties in the layers 0-20 cm and 20-40 cm after plant removal with bulldozers.

Alterations of soil chemical properties by eucalyptus cultivation in five regions in the Rio Doce Valley

Little is currently known about modifications in edaphic characteristics caused by short-rotation eucalyptus and the impacts of these alterations on the sustainability of eucalyptus wood production. This study was carried out to identify theses changes at five sites of eucalyptus plantation in the region of the Rio Doce Valley, state of Minas Gerais, Brazil. Areas with more than three previous eucalyptus cycles, adjacent to pasture land or native forest, were chosen. Soil samples were collected and soil fertility analyzed by routine methods and other fractionation methods in order to measure alterations in the K, Ca and Mg contents as a consequence of eucalyptus cultivation. In the eucalyptus areas, reductions in the exchangeable Ca2+, Mg2+ and K+ contents and pH were observed and increased Al3+ and H + Al contents. Of all nutrients, only P contents (Mehlich-1 P) increased in the eucalyptus areas. The reduction in exchangeable forms and in medium-term soil nutrient pools indicates the need for higher nutrient rates than the currently applied in order to prevent nutritional limitations and soil nutrient exhaustion. After several eucalyptus rotations there was a recovery in the SOM content in comparison to degraded pasture soils, although not to the level of the native forest soil. The positive correlation between effective CEC and medium-term non-exchangeable Ca, Mg and K with SOM emphasizes the need for adequate fertilizer and plant residue management to sustain or even increase forest productivity in future cycles.

Labile and stable fractions of soil organic matter under management systems and native cerrado

Soil organic matter can be analyzed on the basis of the different fractions. Changes in the levels of organic matter, caused by land use, can be better understood by alterations in the different compartments. The aim of this study was to evaluate the effect of different management systems on the labile and stable organic matter of a dystrophic Red Latosol (Oxisol). The following properties were determined: total organic C and total N (TOC and TN), particulate organic C and particulate N (POC and PN), organic C and N mineral-associated (MOC and NM) and particulate organic C associated with aggregate classes (POCA). Eight treatments were used: seven with soil management systems and one with native Cerrado as a reference. The experiment was designed to study the dynamics of systems of tillage and crop rotation, alternating in time and space. The experimental design was a randomized block design with three replications. The soil samples were collected from five depths: 0-5, 5-10, 10-20, 20-30 and 30-40 cm. Changes in organic C by land use occurred mainly in the fraction of particulate organic matter (> 53 mm). Proper management of grazing promoted increased levels of particulate organic matter by association with larger aggregates (2-8 mm), demonstrating the importance of the formation of this aggregate class for C protection in pasture.

Availability of soil water under tillage systems, mulch management and citrus rootstocks

The increased availability of soil water is important for the management of non-irrigated orange orchards. The objective of this study was to evaluate the availability of soil water in a Haplorthox (Rhodic Ferralsol) under different tillage systems used for orchard plantation, mulch management and rootstocks in a "Pêra" orange orchard in northwest Paraná, Brazil. An experiment in a split-split-plot design was established in 2002, in an area cultivated with Brachiaria brizantha grass in which three tillage systems (no tillage, conventional tillage and strip-tillage) were used for orchard plantation. This grass was mowed twice a year between the rows, representing two mulch managements in the split plots (no mulching and mulching in the plant rows). The split-split-plots were represented by two rootstocks ("Rangpur" lime and "Cleopatra" mandarin). The soil water content in the plant rows was evaluated in the 0-20 cm layer in 2007 and at 0-20 and 20-40 cm in 2008-2009. The effect of soil tillage systems prior to implantation of orange orchards on soil water availability was less pronounced than mulching and the rootstocks. The soil water availability was lower when "Pêra" orange trees were grafted on "Cleopatra" mandarin than on "Rangpur" lime rootstocks. Mulching had a positive influence on soil water availability in the sandy surface layer (0-20 cm) and sandy clay loam subsurface (20-40 cm) of the soil in the spring. The production of B. brizantha between the rows and residue disposal in the plant rows as mulch increased water availability to the "Pêra" orange trees.

Temporal variability of soil water content under different surface conditions in the semiarid region of the Pernambuco state

Rainfall in the semiarid region of Pernambuco is characterized by irregular distribution in time and space, which significantly hinders the rainfed agriculture in the region. This work aims to evaluate the temporal profile of soil moisture in the semiarid region of the Pernambuco State (Brazil) and the effect of different soil surface conditions on soil water content variation and the yield of rainfed beans. To monitor soil water content, five plots 4.5 m wide by 11 m long were installed in a Yellow Argisol (Ultisol). The following treatments were adopted in the experimental plots: natural vegetation, bean intercropped with cactus, beans planted down the slope, beans planted along contour lines with mulch and rock barriers, and bare soil. In each plot, eight PVC access tubes were installed for monitoring the soil water content profile at depths of 0.20 and 0.40 m using a neutron probe device. The surface condition significantly influenced the soil water content variation, both in the dry and rainy seasons. The use of mulch, associated with rock barriers, provided higher soil water content levels than the other treatments and increased the rainfed beans production.