Development and validation of PCR-based assays for diagnosis of American cutaneous leishmaniasis and identificatio nof the parasite species

In this study, PCR assays targeting different Leishmania heat-shock protein 70 gene (hsp70) regions, producing fragments ranging in size from 230-390 bp were developed and evaluated to determine their potential as a tool for the specific molecular diagnosis of cutaneous leishmaniasis (CL). A total of 70 Leishmania strains were analysed, including seven reference strains (RS) and 63 previously typed strains. Analysis of the RS indicated a specific region of 234 bp in the hsp70 gene as a valid target that was highly sensitive for detection of Leishmania species DNA with capacity of distinguishing all analyzed species, after polymerase chain reaction-restriction fragment length polymorfism (PCR-RFLP). This PCR assay was compared with other PCR targets used for the molecular diagnosis of leishmaniasis: hsp70 (1400-bp region), internal transcribed spacer (ITS)1 and glucose-6-phosphate dehydrogenase (G6pd). A good agreement among the methods was observed concerning the Leishmania species identification. Moreover, to evaluate the potential for molecular diagnosis, we compared the PCR targets hsp70-234 bp, ITS1, G6pd and mkDNA using a panel of 99 DNA samples from tissue fragments collected from patients with confirmed CL. Both PCR-hsp70-234 bp and PCR-ITS1 detected Leishmania DNA in more than 70% of the samples. However, using hsp70-234 bp PCR-RFLP, identification of all of the Leishmania species associated with CL in Brazil can be achieved employing a simpler and cheaper electrophoresis protocol.

Diversity and distribution of sandflies (Diptera: Psychodidae: Phlebotominae) in a military area in the state of Amazonas, Brazil

This study reports the distribution, ecotopes and fauna diversity of sandflies captured in five training bases on a military reserve in Manaus, state of Amazonas (AM). A total of 10,762 specimens were collected, which were distributed among 58 species, with the highest number recorded at Base Instruction 1 (BI1). A higher rate of species richness was found at the Base Instruction Boina Rajada and low levels of diversity associated with a high abundance index with the clear dominance of Lutzomyia umbratilis, Lutzomyia ruii and Lutzomyia anduzei were found at BI1. The abundance of Lu. umbratilis raises the possibility of outbreaks of American cutaneous leishmaniasis by the main vector of the disease in AM.

Severity of chronic experimental Chagas' heart disease parallels tumour necrosis factor and nitric oxide levels in the serum: models of mild and severe disease

Heart tissue inflammation, progressive fibrosis and electrocardiographic alterations occur in approximately 30% of patients infected by Trypanosoma cruzi, 10-30 years after infection. Further, plasma levels of tumour necrosis factor (TNF) and nitric oxide (NO) are associated with the degree of heart dysfunction in chronic chagasic cardiomyopathy (CCC). Thus, our aim was to establish experimental models that mimic a range of parasitological, pathological and cardiac alterations described in patients with chronic Chagas’ heart disease and evaluate whether heart disease severity was associated with increased TNF and NO levels in the serum. Our results show that C3H/He mice chronically infected with the Colombian T. cruzi strain have more severe cardiac parasitism and inflammation than C57BL/6 mice. In addition, connexin 43 disorganisation and fibronectin deposition in the heart tissue, increased levels of creatine kinase cardiac MB isoenzyme activity in the serum and more severe electrical abnormalities were observed in T. cruzi-infected C3H/He mice compared to C57BL/6 mice. Therefore, T. cruzi-infected C3H/He and C57BL/6 mice represent severe and mild models of CCC, respectively. Moreover, the CCC severity paralleled the TNF and NO levels in the serum. Therefore, these models are appropriate for studying the pathophysiology and biomarkers of CCC progression, as well as for testing therapeutic agents for patients with Chagas’ heart disease.

Development and validation of the Family Vulnerability Index to Disability and Dependence (FVI-DD)

This exploratory, descriptive, cross-sectional, and quantitative study aimed to develop and validate an index of family vulnerability to disability and dependence (FVI-DD). This study was adapted from the Family Development Index, with the addition of social and health indicators of disability and dependence. The instrument was applied to 248 families in the city of Sao Paulo, followed by exploratory factor analysis. Factor validation was performed using the concurrent and discriminant validity of the Lawton scale and Katz Index. The descriptive level adopted for the study was p < 0.05. The final vulnerability index comprised 50 questions classified into seven factors contemplating social and health dimensions, and this index exhibited good internal consistency (Cronbach’s alpha = 0.82). FVI-DD was validated using both the Lawton scale and Katz Index. We conclude that FVI-DD can accurately and reliably assess family vulnerability to disability and dependence.

Development and validation of an instrument for evaluating the ludicity of games in health education

Abstract OBJECTIVE Developing and validating an instrument to evaluate the playfulness of games in health education contexts. METHODOLOGY A methodological, exploratory and descriptive research, developed in two stages: 1. Application of an open questionnaire to 50 graduate students, with content analysis of the answers and calculation of Kappa coefficient for defining items; 2. Procedures for construction of scales, with content validation by judges and analysis of the consensus estimate byContent Validity Index(CVI). RESULTS 53 items regarding the restless character of the games in the dimensions of playfulness, the formative components of learning and the profiles of the players. CONCLUSION Ludicity can be assessed by validated items related to the degree of involvement, immersion and reinvention of the subjects in the game along with the dynamics and playability of the game.

Density and distribution of nests of Mycetophylax simplex (Emery) (Hymenoptera, Formicidae) in areas with mobile dunes on the northern coast of Rio Grande do Sul, Brazil

Studies on lower attines are scarce, especially on nesting and foraging ecology and behavior. This study aimed to contribute to the knowledge of an Attini in dunes ecosystems through the description of density and spatial distribution of Mycetophylax simplex (Emery, 1887) nests in a strip of mobile dunes in the Praia Grande beach, Torres, northern coast of Rio Grande do Sul, Brazil. The density and spatial distribution of nests were estimated in four plots of 2,500 m² each, in which were found 20, 209, 284 and 324 nests, with average densities of 0.01 nests/m², 0.09, 0.11 and 0.13 nests/m², respectively. The nests were found near to the vegetation and showed clumped distribution. The density and distribution pattern of the nests seem to be related to the availability of nesting places and foraging resources.

Systematics and distribution of the genus Johnsonius Marsh (Hymenoptera, Braconidae, Doryctinae) with description of two new species from Brazil

The genus Johnsonius Marsh includes five species, most of them known from Costa Rica and one known from Venezuela, Peru and Costa Rica. Two of these species and two new species were recently collected in Atlantic rain forest. Description of the new species, illustrations of some morphological characters, a key for identification of all known species and the new geographic records are presented.

The three phases of the ensemble forecasting of niche models: geographic range and shifts in climatically suitable areas of Utetheisa ornatrix (Lepidoptera, Arctiidae)

Species' geographic ranges are usually considered as basic units in macroecology and biogeography, yet it is still difficult to measure them accurately for many reasons. About 20 years ago, researchers started using local data on species' occurrences to estimate broad scale ranges, thereby establishing the niche modeling approach. However, there are still many problems in model evaluation and application, and one of the solutions is to find a consensus solution among models derived from different mathematical and statistical models for niche modeling, climatic projections and variable combination, all of which are sources of uncertainty during niche modeling. In this paper, we discuss this approach of ensemble forecasting and propose that it can be divided into three phases with increasing levels of complexity. Phase I is the simple combination of maps to achieve a consensual and hopefully conservative solution. In Phase II, differences among the maps used are described by multivariate analyses, and Phase III consists of the quantitative evaluation of the relative magnitude of uncertainties from different sources and their mapping. To illustrate these developments, we analyzed the occurrence data of the tiger moth, Utetheisa ornatrix (Lepidoptera, Arctiidae), a Neotropical moth species, and modeled its geographic range in current and future climates.

Observations on the biology and distribution of Uresiphita reversalis (Lepidoptera, Crambidae), a defoliator of the native tree Calia secundiflora in México

Observations on the biology and distribution of Uresiphita reversalis (Lepidoptera, Crambidae), a defoliator of the native tree Calia secundiflora in México. Uresiphita reversalis (Guenée, 1854) feeding on Calia secundiflora (Ortega) Yakovlev is recorded for the first time in the state of Hidalgo, Mexico. New aspects regarding the life cycle, feeding behaviour, geographical distribution and host plant damage by U. reversalis on C. secundiflora are here presented and discussed.

Interaction between Tephritidae (Insecta, Diptera) and plants of the family Asteraceae: new host and distribution records for the state of Rio Grande do Sul, Brazil

Twenty species of Tephritidae (Diptera) are recorded in association with capitula of plants in the family Asteraceae. The Tephritidae genus Tetreuarestais registered for Rio Grande do Sul for the first time. Five species of Tephritidae are newly recorded for Rio Grande do Sul, and new hosts are recorded for the following fly species: Dioxyna chilensis (Macquart), Plaumannimyia dolores (Hering), Plaumannimyia imitatrix (Hering), Plaumannimyia miseta(Hering), Plaumannimyia pallensHering, Tomoplagia incompleta (Williston), Tomoplagia matzenbacheriPrado, Norrbom & Lewinsohn, Tomoplagia reimoseriHendel, Xanthaciura biocellata(Thomson) and Xanthaciura chrysura(Thomson).

Calibration of the century, apsim and ndicea models of decomposition and n mineralization of plant residues in the humid tropics

The aim of this study was to calibrate the CENTURY, APSIM and NDICEA simulation models for estimating decomposition and N mineralization rates of plant organic materials (Arachis pintoi, Calopogonium mucunoides, Stizolobium aterrimum, Stylosanthes guyanensis) for 360 days in the Atlantic rainforest bioma of Brazil. The models´ default settings overestimated the decomposition and N-mineralization of plant residues, underlining the fact that the models must be calibrated for use under tropical conditions. For example, the APSIM model simulated the decomposition of the Stizolobium aterrimum and Calopogonium mucunoides residues with an error rate of 37.62 and 48.23 %, respectively, by comparison with the observed data, and was the least accurate model in the absence of calibration. At the default settings, the NDICEA model produced an error rate of 10.46 and 14.46 % and the CENTURY model, 21.42 and 31.84 %, respectively, for Stizolobium aterrimum and Calopogonium mucunoides residue decomposition. After calibration, the models showed a high level of accuracy in estimating decomposition and N- mineralization, with an error rate of less than 20 %. The calibrated NDICEA model showed the highest level of accuracy, followed by the APSIM and CENTURY. All models performed poorly in the first few months of decomposition and N-mineralization, indicating the need of an additional parameter for initial microorganism growth on the residues that would take the effect of leaching due to rainfall into account.

Evaluation of statistical and geostatistical models of digital soil properties mapping in tropical mountain regions

Soil properties have an enormous impact on economic and environmental aspects of agricultural production. Quantitative relationships between soil properties and the factors that influence their variability are the basis of digital soil mapping. The predictive models of soil properties evaluated in this work are statistical (multiple linear regression-MLR) and geostatistical (ordinary kriging and co-kriging). The study was conducted in the municipality of Bom Jardim, RJ, using a soil database with 208 sampling points. Predictive models were evaluated for sand, silt and clay fractions, pH in water and organic carbon at six depths according to the specifications of the consortium of digital soil mapping at the global level (GlobalSoilMap). Continuous covariates and categorical predictors were used and their contributions to the model assessed. Only the environmental covariates elevation, aspect, stream power index (SPI), soil wetness index (SWI), normalized difference vegetation index (NDVI), and b3/b2 band ratio were significantly correlated with soil properties. The predictive models had a mean coefficient of determination of 0.21. Best results were obtained with the geostatistical predictive models, where the highest coefficient of determination 0.43 was associated with sand properties between 60 to 100 cm deep. The use of a sparse data set of soil properties for digital mapping can explain only part of the spatial variation of these properties. The results may be related to the sampling density and the quantity and quality of the environmental covariates and predictive models used.

Development and fruit yield of strawberry plants as affected by crown diameter and plantlet growing period

The objective of this work was to evaluate the influence of crown diameter and growing period of runner tips in 128 cell-trays on growth and yield of strawberry in the field. Treatments consisted of three classes of runner tip crown diameters, between 2.0 to 3.9 mm; 4.0 to 5.5 mm and 5.6 to 7.0 mm, respectively, and four growing periods in trays, 24; 39; 54 and 69 days, respectively. Higher shoot dry mass of transplants at planting and earlier yield of plants in the field were obtained in transplants grown for 69 days in trays. Larger runner tips lead to more vigorous transplants at planting and plants with higher vegetative growth in the field, with minor impact on yield. Increasing the growing time of runner tips in trays improved early fruit yield and decreased plant vegetative growth in the field.

Leaf development and growth of selected sugarcane clones in a subtropical environment

The objective of this work was to compare phyllochron and leaf area on individual stalks of selected sugarcane (Saccharum officinarum) clones grown in a subtropical environment. A two-year field experiment was carried out in Santa Maria, RS, Brazil, during the 2007/2008 and 2008/2009 growing seasons. Three sugarcane clones were used: IAC 822045 (early), SP 711406 (median) and CB 4176 (late), in a complete randomized block design, with four replications. Expanded leaf and total leaf (tips) number were determined, and expanded leaf area was measured. The phyllochron (ºC day leaf-1) based on expanded and tip leaf number was estimated, using 10ºC as base temperature. Allometric relationships between individual area and number of expanded leaves were fitted. Phyllochron was analysed as a four-factor experiment: clone, growing season, phyllochron phase, and phyllochron type. The early clone had the greatest potential for growing in a subtropical climate because of higher leaf output and leaf area.

Gene expression profile during coffee fruit development and identification of candidate markers for phenological stages

The objective of this work was to identify genes that could be used as suitable markers for molecular recognition of phenological stages during coffee (Coffea arabica) fruit development. Four cultivars were evaluated as to their differential expression of genes associated to fruit development and maturation processes. Gene expression was characterized by both semi-quantitative and quantitative RT-PCR, in fruit harvested at seven different developmental stages, during three different seasons. No size polymorphisms or differential expression were observed among the cultivars for the evaluated genes; however, distinct expression profiles along fruit development were determined for each gene. Four out of the 28 evaluated genes exhibited a regular expression profile in all cultivars and harvest seasons, and, therefore, they were validated as candidate phenological markers of coffee fruit. The gene α-galactosidase can be used as a marker of green stage, caffeine synthase as a marker of transition to green and yellowish-green stages, and isocitrate lyase and ethylene receptor 3 as markers of late maturation.