A simplified method for sample collection and DNA isolation for polymerase chain reaction detection of Trypanosoma rangeli and Trypanosoma cruzi in triatomine vectors

Due to the overlapping distribution of Trypanosoma rangeli and T. cruzi in Central and South America, sharing several reservoirs and triatomine vectors, we herein describe a simple method to collect triatomine feces and hemolymph in filter paper for further detection and specific characterization of these two trypanosomes. Experimentally infected triatomines feces and hemolymph were collected in filter paper and specific detection of T. rangeli or T. cruzi DNA by polymerase chain reaction was achieved. This simple DNA collection method allows sample collection in the field and further specific trypanosome detection and characterization in the laboratory.

Sylvatic population of Triatoma infestans from the Bolivian Chaco: from field collection to characterization

A sylvatic Triatoma infestans DM (dark morph) population detected in the Bolivian Chaco was characterized and compared with various domestic ones. The degree of differentiation of DM was clearly within the T. infestans intra-specific level. Nevertheless marked chromatic and morphometric differences as well as differences in antennal pattern, chromosome banding and randomly amplified polymorphic DNA support the hypothesis of a distinct population. Continuous exchange of insects between wild and domestic habitats seems unlikely in the Chaco.

Identification of a genetic marker associated with the resistance to Schistosoma mansoni infection using random amplified polymorphic DNA analysis

In schistosomiasis, the host/parasite interaction remains not completely understood. Many questions related to the susceptibility of snails to infection by respective trematode still remain unanswered. The control of schistosomiasis requires a good understanding of the host/parasite association. In this work, the susceptibility/resistance to Schistosoma mansoni infection within Biomphalaria alexandrina snails were studied starting one month post infection and continuing thereafter weekly up to 10 weeks after miracidia exposure. Genetic variations between susceptible and resistant strains to Schistosoma infection within B. alexandrina snails using random amplified polymorphic DNA analysis technique were also carried out. The results showed that 39.8% of the examined field snails were resistant, while 60.2% of these snails showed high infection rates.In the resistant genotype snails, OPA-02 primer produced a major low molecular weight marker 430 bp. Among the two snail strains there were interpopulational variations, while the individual specimens from the same snail strain, either susceptible or resistant, record semi-identical genetic bands. Also, the resistant character was ascendant in contrast to a decline in the susceptibility of snails from one generation to the next.

Quantification of the genetic change in the transition of Rhodnius pallescens Barber, 1932 (Hemiptera: Reduviidae) from field to laboratory

Previous studies have reported genetic differences between wild-caught sylvatic, domestic and laboratory pop-ulations of several Triatominae species. The differences between sylvatic and laboratory colonies parallel are similar to the differences observed between sylvatic and domestic populations. Laboratory colonies are frequently used as references for field populations, but the consequences of founder events on the genetic makeup of laboratory or domestic populations are rarely quantified. Our goal was to quantify the genetic change in Rhodnius pallescens populations artificially submitted to founder effects via laboratory colonization. We compared the genetic makeup of two sylvatic populations and their laboratory descendants using a panel of 10 microsatellite markers. Both sylvatic populations were initially collected from palm trees, but the colonies differed in the number of founder insects and amount of time kept in the laboratory. We evaluated allelic polymorphism, differences between expected and observed heterozygosity, estimates of population differentiation (Fst) and inbreeding (Fis, Fit) and cluster analyses based on Nei's distances. We found a unique genetic structure for each sample population, with significant differentiation between the field insects and each of the laboratory generations. These analyses showed strong founder effects and showed that genetic drift had led to a genetic equilibrium over several generations of isolation. Our results suggest that laboratory colonies of R. pallescens have a different genetic structure than their wild relatives and similar processes likely affect other Triatominae laboratory stocks.

Genetic variability in G2 and F2 region between biological clones of human respiratory syncytial virus with or without host immune selection pressure

Human respiratory syncytial virus (HRSV) is an important respiratory pathogens among children between zero-five years old. Host immunity and viral genetic variability are important factors that can make vaccine production difficult. In this work, differences between biological clones of HRSV were detected in clinical samples in the absence and presence of serum collected from children in the convalescent phase of the illness and from their biological mothers. Viral clones were selected by plaque assay in the absence and presence of serum and nucleotide sequences of the G2 and F2 genes of HRSV biological clones were compared. One non-synonymous mutation was found in the F gene (Ile5Asn) in one clone of an HRSV-B sample and one non-synonymous mutation was found in the G gene (Ser291Pro) in four clones of the same HRSV-B sample. Only one of these clones was obtained after treatment with the child's serum. In addition, some synonymous mutations were determined in two clones of the HRSV-A samples. In conclusion, it is possible that minor sequences could be selected by host antibodies contributing to the HRSV evolutionary process, hampering the development of an effective vaccine, since we verify the same codon alteration in absence and presence of human sera in individual clones of BR-85 sample.

Mapping the nursing care with the NIC for patients in risk for pressure ulcer

Objective:To identify the nursing care prescribed for patients in risk for pressure ulcer (PU) and to compare those with the Nursing Interventions Classification (NIC) interventions. Method: Cross mapping study conducted in a university hospital. The sample was composed of 219 adult patients hospitalized in clinical and surgical units. The inclusion criteria were: score ≤ 13 in the Braden Scale and one of the nursing diagnoses, Self-Care deficit syndrome, Impaired physical mobility, Impaired tissue integrity, Impaired skin integrity, Risk for impaired skin integrity. The data were collected retrospectively in a nursing prescription system and statistically analyzed by crossed mapping. Result: It was identified 32 different nursing cares to prevent PU, mapped in 17 different NIC interventions, within them: Skin surveillance, Pressure ulcer prevention and Positioning. Conclusion: The cross mapping showed similarities between the prescribed nursing care and the NIC interventions.

Effectiveness of Enteral Nutritional Therapy in the Healing Process of Pressure Ulcers: A Systematic Review

OBJECTIVE To evaluate the effectiveness of enteral nutritional therapy (ENT) in the healing process of pressure ulcers (PU) in adults and the elderly. METHOD A systematic review whose studies were identified through the databases of Cochrane, MEDLINE/PubMed, SciELO, LILACS, EMBASE, CINAHL, Web of Science, and manual searches. It included randomized clinical trials (RCTs) without delimiting the period or language of publication, which addressed adults and elderly patients with pressure ulcers in a comparative treatment of enteral nutritional therapy and placebo or between enteral nutritional therapy with different compositions and dosages. RESULTS We included ten studies that considered different interventions. It resulted in more pressure ulcers healed in the groups that received the intervention. The included studies were heterogeneous with regard to patients, the type of intervention, the sample and the follow-up period, all of which made meta-analysis impossible. CONCLUSION Although the enteral nutritional therapy demonstrates a promotion of pressure ulcer healing, sufficient evidence to confirm the hypothesis was not found.

Scaling of semivariograms and the kriging estimation of field-measured properties

Two methods were evaluated for scaling a set of semivariograms into a unified function for kriging estimation of field-measured properties. Scaling is performed using sample variances and sills of individual semivariograms as scale factors. Theoretical developments show that kriging weights are independent of the scaling factor which appears simply as a constant multiplying both sides of the kriging equations. The scaling techniques were applied to four sets of semivariograms representing spatial scales of 30 x 30 m to 600 x 900 km. Experimental semivariograms in each set successfully coalesced into a single curve by variances and sills of individual semivariograms. To evaluate the scaling techniques, kriged estimates derived from scaled semivariogram models were compared with those derived from unscaled models. Differences in kriged estimates of the order of 5% were found for the cases in which the scaling technique was not successful in coalescing the individual semivariograms, which also means that the spatial variability of these properties is different. The proposed scaling techniques enhance interpretation of semivariograms when a variety of measurements are made at the same location. They also reduce computational times for kriging estimations because kriging weights only need to be calculated for one variable. Weights remain unchanged for all other variables in the data set whose semivariograms are scaled.

An improved method for measuring soil microbial activity by gas phase flow injection analysis

The rate of carbon dioxide production is commonly used as a measure of microbial activity in the soil. The traditional method of CO2 determination involves trapping CO2 in an alkali solution and then determining CO2 concentration indirectly by titration of the remaining alkali in the solution. This method is still commonly employed in laboratories throughout the world due to its relative simplicity and the fact that it does not require expensive, specific equipment. However, there are several drawbacks: the method is time-consuming, requires large amounts of chemicals and the consistency of results depends on the operator's skills. With this in mind, an improved method was developed to analyze CO2 captured in alkali traps, which is cheap and relatively simple, with a substantially shorter sample handling time and reproducibility equivalent to the traditional titration method. A comparison of the concentration values determined by gas phase flow injection analysis (GPFIA) and titration showed no significant difference (p > 0.05), but GPFIA has the advantage that only a tenth of the sample volume of the titration method is required. The GPFIA system does not require the purchase of new, costly equipment but the device was constructed from items commonly found in laboratories, with suggestions for alternative configurations for other detection units. Furthermore, GPFIA for CO2 analysis can be equally applied to samples obtained from either the headspace of microcosms or from a sampling chamber that allows CO2 to be released from alkali trapping solutions. The optimised GPFIA method was applied to analyse CO2 released from degrading hydrocarbons from a site contaminated by diesel spillage.

The efficiency of different estimation methods of hydro-physical limits

The soil water available to crops is defined by specific values of water potential limits. Underlying the estimation of hydro-physical limits, identified as permanent wilting point (PWP) and field capacity (FC), is the selection of a suitable method based on a multi-criteria analysis that is not always clear and defined. In this kind of analysis, the time required for measurements must be taken into consideration as well as other external measurement factors, e.g., the reliability and suitability of the study area, measurement uncertainty, cost, effort and labour invested. In this paper, the efficiency of different methods for determining hydro-physical limits is evaluated by using indices that allow for the calculation of efficiency in terms of effort and cost. The analysis evaluates both direct determination methods (pressure plate - PP and water activity meter - WAM) and indirect estimation methods (pedotransfer functions - PTFs). The PTFs must be validated for the area of interest before use, but the time and cost associated with this validation are not included in the cost of analysis. Compared to the other methods, the combined use of PP and WAM to determine hydro-physical limits differs significantly in time and cost required and quality of information. For direct methods, increasing sample size significantly reduces cost and time. This paper assesses the effectiveness of combining a general analysis based on efficiency indices and more specific analyses based on the different influencing factors, which were considered separately so as not to mask potential benefits or drawbacks that are not evidenced in efficiency estimation.

USE OF SCALED SEMIVARIOGRAMS IN THE PLANNING SAMPLE OF SOIL CHEMICAL PROPERTIES IN SOUTHERN AMAZONAS, BRAZIL

The lack of information concerning the variability of soil properties has been a major concern of researchers in the Amazon region. Thus, the aim of this study was to evaluate the spatial variability of soil chemical properties and determine minimal sampling density to characterize the variability of these properties in five environments located in the south of the State of Amazonas, Brazil. The five environments were archaeological dark earth (ADE), forest, pasture land, agroforestry operation, and sugarcane crop. Regular 70 × 70 m mesh grids were set up in these areas, with 64 sample points spaced at 10 m distance. Soil samples were collected at the 0.0-0.1 m depth. The chemical properties of pH in water, OM, P, K, Ca, Mg, H+Al, SB, CEC, and V were determined at these points. Data were analyzed by descriptive and geostatistical analyses. A large part of the data analyzed showed spatial dependence. Chemical properties were best fitted to the spherical model in almost all the environments evaluated, except for the sugarcane field with a better fit to the exponential model. ADE and sugarcane areas had greater heterogeneity of soil chemical properties, showing a greater range and higher sampling density; however, forest and agroforestry areas had less variability of chemical properties.

Genetic diversity and virulence pattern in field populations of Pyricularia grisea from rice cultivar Metica-1

Rice blast is a major yield constraint of the irrigated rice in the State of Tocantins, Brazil. The objective of this investigation was to study the phenotypic and genetic diversity within the pathogen population of Pyricularia grisea in samples collected from four individual farms of rice cultivar Metica-1, under epidemic conditions of leaf blast. A set of 87 isolates was tested on 32 rice genotypes including eight international differentials. Considering 80% similarity in virulence, two groups comprising a total of 81 isolates were recognized, independently of the farms from which they were collected. Eighty percent of the isolates pertained to pathotype ID-14, indicating high cultivar specificity and narrow diversity of virulence in the sample population. The virulence in pathogen population on rice cultivars BR-IRGA 409 and Rio Formoso was low. Analysis of P. grisea isolates using rep-PCR with two primer sequences from Pot2 generated fingerprint profiles of one to nine bands. Cluster analysis revealed the occurrence of six fingerprint groups with similarities ranging from 0.09 to 1. There was no straight relationship between virulence of the isolates based on reaction pattern on 32 genotypes and grouping based on Pot2 rep-PCR analysis of P. grisea isolates collected from 'Metica-1'.

Solar radiation use efficiency by soybean under field conditions in the Amazon region

The objective of this work was to evaluate the efficiency of soybean (Glycine max) in intercepting and using solar radiation under natural field conditions, in the Amazon region, Brazil. The meteorological data and the values of soybean growth and leaf area were obtained from an agrometeorological experiment carried out in Paragominas, Pará state, during 2007 and 2008. The radiation use efficiency (RUE) was obtained from the ratio between the above-ground biomass production and the intercepted photosynthetically active radiation (PAR) accumulated to 99 and 95 days after sowing, in 2007 and 2008, respectively. Climatic conditions during the experiment were very distinct, with reduction in rainfall in 2007, which began during the soybean mid-cycle, due to the El Niño phenomenon. An important reduction in the leaf area index and biomass production was observed during 2007. Under natural field conditions in the Amazon region, the values of RUE were 1.46 and 1.99 g MJ-1 PAR in the 2007 and 2008 experiments, respectively. The probable reason for the differences found between these years might be associated to the water restriction in 2007 coupled with the higher air temperature and vapor pressure deficit, and also to the increase in the fraction of diffuse radiation that reached the land surface in 2008.

Mecanização no preparo de amostras por microondas: o estado da arte

This review presents the latest advances of the mechanisation in sample preparation using microwave oven. The fundamental principles of microwaves field interaction with matter are presented, followed by the basic principles of microwave equipment construction. Hyphenation of microwave oven with flow injection analysis or robotic are discussed as well as some metodological difficulties and solutions by its implementation.

Automated flow-injection method for cadmium determination with pre-concentration and reagent preparation on-line

The spectrophotometric determination of Cd(II) using a flow injection system provided with a solid-phase reactor for cadmium preconcentration and on-line reagent preparation, is described. It is based on the formation of a dithizone-Cd complex in basic medium. The calibration curve is linear between 6 and 300 µg L-1 Cd(II), with a detection limit of 5.4 µg L-1, an RSD of 3.7% (10 replicates in duplicate) and a sample frequency of 11.4 h-1. The proposed method was satisfactorily applied to the determination of Cd(II) in surface, well and drinking waters.