HIGH SENSITIVE PCR METHOD FOR DETECTION OF PATHOGENIC Leptospira spp. IN PARAFFIN-EMBEDDED TISSUES

This study describes the development and application of a new PCR assay for the specific detection of pathogenic leptospires and its comparison with a previously reported PCR protocol. New primers were designed for PCR optimization and evaluation in artificially-infected paraffin-embedded tissues. PCR was then applied to post-mortem, paraffin-embedded samples, followed by amplicon sequencing. The PCR was more efficient than the reported protocol, allowing the amplification of expected DNA fragment from the artificially infected samples and from 44% of the post-mortem samples. The sequences of PCR amplicons from different patients showed >99% homology with pathogenic leptospires DNA sequences. The applicability of a highly sensitive and specific tool to screen histological specimens for the detection of pathogenic Leptospira spp. would facilitate a better assessment of the prevalence and epidemiology of leptospirosis, which constitutes a health problem in many countries.

Detection of Leishmania braziliensis in human paraffin-embedded tissues from Tucumán, Argentina by polymerase chain reaction

American cutaneous leishmaniasis (ACL) is an endemic disease in Northern Argentina. We applied the polymerase chain reaction (PCR) followed by a hybridization labelled probe to 21 paraffin embedded human skin biopsies, already analyzed histologically, from leishmaniasis endemic areas in the province of Tucumán, Argentina. We used primers previously designed to detect a Leishmania-specific 120-base-pair fragment of kinetoplast DNA minicircle, other two primer pairs that amplify kDNA minicircles belonging to the L. braziliensis and L. mexicana complexes respectively, and specific oligonucleotide primers to detect L. (V.) braziliensis which amplify the sequence of the ribosomal protein L-14 of this species. The PCR-hybridization showed a sensitivity of 90.5% when compared to the histopathology test which was 61.9%. Five of the total samples analyzed were positive for the L. braziliensis complex whilst none was positive for the L. mexicana complex. The specific primers for L. (V.) braziliensis detected the parasite in four samples. These results are consistent with those reported for close endemic areas and demonstrate that the causative agent of human leishmaniasis in the analyzed cases was L. (V.) braziliensis. PCR should be used as a diagnostic tool for tegumentary leishmaniasis, especially in the mucosal form, and as a valuable technique for the identification of the Leishmania species that causes the disease in certain areas.

Demographic determinants of survival of people living with HIV attending an outpatient reference unit in the city of Três Lagoas, state of Mato Grosso do Sul, Brazil, in the period 1984-2009

INTRODUCTION: While the incidence of HIV infection and AIDS is increasing in small Brazilian cities, epidemiological studies are often conducted in large urban centers. METHODS: Our group conducted a retrospective analysis of survival determinants among 358 patients who attended a reference unit in a small city. RESULTS: Death risk was lower among men that had sex with men, patients with an HIV-seropositive partner, and those admitted after highly active antiretroviral therapy (HAART) was available. CONCLUSIONS: The study documents the striking beneficial effect of HAART. The finding of other groups with improved survival may aid in the development of programmatic strategies.

Female gender but not season of birth is associated with mood seasonality in a near-equatorial Brazilian city

OBJECTIVE: Studies conducted mainly in countries located in the Northern Hemisphere have shown that season of birth influences mood seasonality. Greater mood seasonality has been observed for individuals born during spring/summer months than those born during autumn/winter months. Expanding past research to the Southern Hemisphere, in this study we examine the influence of season of birth on mood seasonality in a sample of 1,247 healthy young Brazilians. METHOD: The Seasonal Pattern Assessment Questionnaire was used to compute a global seasonality score as a measure of mood seasonality in a cross-sectional study. RESULTS: Analysis of covariance was conducted to examine the effects of month of birth and gender on mood seasonality, with age entered as a covariate. A main effect of gender was observed, F (1, 1197) = 17.86, p < .01; partial Eta-squared = .02, with mood seasonality being higher for females (M = 8) than for males (M = 7). Contradicting previous findings, no significant main effect for month of birth was observed, F (1, 1197) = 0.65, p > .05. CONCLUSION: The unexpected finding is tentatively explained by differences in geographic location and weather fluctuations between the sampling location in Brazil and other countries where season of birth has been found to influence mood seasonality. Additional studies with larger samples from the Southern Hemisphere are necessary to shed additional light on the possible significant influence of season of birth on mood.

Three new species of Mesabolivar (Aranea, Pholcidae) from leaf litter in urban environments in the city of São Paulo, São Paulo, Brazil

In this study we describe three new litter inhabiting species of Mesabolivar González-Sponga, 1998 from nine urban forest remnants in the metropolitan region of the city of São Paulo, Brazil: M. forceps, M. mairyara and M. cavicelatus. In three of these remnants, we conduced a three year sampling using pitfall traps. Mesabolivar forceps sp. nov. was the most abundant pholcid (n=273 adults), always present in the samples, but with highest numbers in spring and summer. Mesabolivar mairyara sp. nov. was the second most abundant species (n=32), but the majority of individuals were collected in March 2001. Only three individuals of M. cavicelatus sp. nov. were collected.

Diversity of Campylobacter Isolates from Three Activated Sludge Systems

Thermophilic campylobacters were isolated from three sewage plants in Rio de Janeiro, RJ, Brazil and identified. Laboratory analysis of 390 sewage samples showed the presence of 169 thermophilic strains. The results demonstrated that human and animal pathogenic biotypes could be isolated from activated sludge during the initial processing steps. The aeration tank could be considered a barrier to Campylobacter survival. C. jejuni was the prevalent species isolated (40.8%).The most common biotypes were C. jejuni biotype I (21.3%), C. coli biotype I (16%) and C. jejuni biotype II ( 14.8%).

Clinical and epidemiological aspects of human parvovirus B19 infection in an urban area in Brazil (Niterói city area, State of Rio de Janeiro, Brazil)

This study was designed to analyse the clinical and epidemiological data from human parvovirus B19 cases in a six-year study of rash diseases conduct in an urban area in Brazil (Niterói city area, State of Rio de Janeiro). A total of 673 patients with acute rash diseases were seen at two primary health care units and at a general hospital. A clotted blood sample was collected from all subjects at the time of consultation. Forty-nine per cent (330 cases) of the patients were negative for dengue, rubella and measles IgM or for low avidity IgG to HHV-6. Of these 330, 105 (31.8%) were identified as IgM positive to parvovirus B19 by using an antibody capture EIA. During the study period, three distinct peaks of parvovirus infection were detected, suggesting that the disease appears to cycle in approximately 4-5 years. B19 infection was characterized by variable combinations of fever, flu-like symptoms, arthropathy, and gastrointestinal symptoms. Frequency of fever and arthropathy was substantially higher in adults, 75% [chi2 (1 D.F.) = 11.39, p = 0.0007] and 62.5% [chi2 (1 D.F.) = 29.89, p = 0.0000], respectively. "Slapped-cheek" appearance and reticular or lace-like rash were seen in only 30.1% of the children. No adult presented this typical rash. The lack of the typical rash pattern in a large proportion of parvovirus B19 and the similarity of clinical manifestations to other rash diseases, specially to rubella, highlight the difficulty of diagnosing B19 infection on clinical grounds alone.

SPATIAL CORRELATION BETWEEN PHYSICAL PROPERTIES OF SOIL AND WEEDS IN TWO MANAGEMENT SYSTEMS

The spatial correlation between soil properties and weeds is relevant in agronomic and environmental terms. The analysis of this correlation is crucial for the interpretation of its meaning, for influencing factors such as dispersal mechanisms, seed production and survival, and the range of influence of soil management techniques. This study aimed to evaluate the spatial correlation between the physical properties of soil and weeds in no-tillage (NT) and conventional tillage (CT) systems. The following physical properties of soil and weeds were analyzed: soil bulk density, macroporosity, microporosity, total porosity, aeration capacity of soil matrix, soil water content at field capacity, weed shoot biomass, weed density, Commelina benghalensis density, and Bidens pilosa density. Generally, the ranges of the spatial correlations were higher in NT than in CT. The cross-variograms showed that many variables have a structure of combined spatial variation and can therefore be mapped from one another by co-kriging. This combined variation also allows inferences about the physical and biological meanings of the study variables. Results also showed that soil management systems influence the spatial dependence structure significantly.

Yield and fruit quality of apple from conventional and organic production systems

The objective of this study was to assess the yield and fruit quality of apple produced with a conventional and an organic production systems in Southern Brazil. The orchards consisted of alternate rows from 10 to 12-year old 'Royal Gala' and 'Fuji' apple trees on M.7 rootstocks, grown as slender spindles, on 4x6 m spacing. Eighteen apple trees of each cultivar and management system were randomly selected and assessed for nutrition, flowering, fruit set, yield, and fruit quality during two growing seasons (2002/2003 and 2003/2004). The organic management system resulted in lower concentrations of K, Mg, and N in leaves and fruits, and in smaller fruits for both cultivars, and lower fruit yield for 'Fuji' than from the conventional production system. For both cultivars, fruits from the organic orchard harvested at commercial maturity had a more yellowish skin background color, higher percentage of blush in the fruit skin, higher soluble solids content, higher density, higher flesh firmness, and higher severity of russet than fruits from the conventional orchard. Fruit from the organic orchard had lower titratable acidity in 'Royal Gala', and higher incidence of moldy core and lower incidence of watercore in 'Fuji', than fruit from the conventional orchard. A non-trained sensory panel detected no significant differences for fruit attributes of taste, flavor and texture between fruit from the production systems for either cultivar.

Bioindicators of soil quality in coffee organic cultivation systems

The objective of this work was to assess the effect of different coffee organic cultivation systems on chemical and biological soil characteristics, in different seasons of the year. The following systems were evaluated: coffee intercropped with one (CJ1), two (CJ2) or three (CJ3) pigeon pea (Cajanus cajan) alleys; coffee planted under full sun (CS); area planted with sweet pepper and snap bean in a conventional tillage system (AC); and secondary forest area (FFR). Row spacing in CJ1, CJ2, CJ3 and CS was 2.0x1.0, 2.8x1.0, 3.6x1.0, and 2.8x1.0 m, respectively. Soil samples were collected at 10-cm depth, during the four seasons of the year. The results were subjected to analysis of variance, principal component analysis, and redundancy analysis. There was an increase in edaphic macrofauna, soil basal respiration, and microbial quotient in the summer. Total macrofauna density was greater in CJ2 followed by CJ3, CS, CJ1, AC and FFR; Coleoptera, Formicidae, and Isoptera were the most abundant groups. There are no significant differences among the areas for soil basal respiration, and the metabolic quotient is higher in CJ1, CJ3, and FFR. Microbial biomass carbon and the contents of K, pH, Ca+Mg, and P show greater values in AC.

Spatial variability of soil physical properties in two management systems in sugarcane crop

The aim of this study was to characterize the spatial variability of soil bulk density (Bd), soil moisture content (θ) and total porosity (Tp) in two management systems of sugarcane harvesting, with or without burning, in a Haplustox soil, in the 0-0.20 m layer. The study area is located in Rio Brilhante, state of Mato Grosso do Sul, Brazil, in Eldorado Sugar Mill. The plots have presented 180 m length, and 145.6 m width, totaling 90 points distributed in the form of a grid of nine rows by ten columns, with points spaced 20 m from its neighbor. Soil samples were collected at 0-0.20 m layer in 2007/2008 and 2008/2009 crops. The harvest with burning system had a higher density compared to mechanized harvest, in the two study periods. The moisture content as well as the porosity increased proportionally with the decrease of the density of the harvest burning system compared to the mechanized.

Economic viability of retrofiting emitters in center pivot irrigation systems

Although several studies have been conducted to evaluate the uniformity of water application under center pivot irrigation systems, there are few studies concerning the economic perspective of such coefficient. The aim of this study is to present a methodology to accomplish an economic analysis as support for the decision-making to retrofit emitters in center pivot irrigation systems, and to attribute an economic meaning to the uniformity coefficient of water application taking into account the response function productivity to the amount of water applied and the sale price of the crops. In the hypothetic calculation example considering the variation of revenue of potato crop under center pivot irrigation system, it was verified that the area with uniformity coefficient of water application of 90% brought an income increase of BR$ 1,992.00, considering an area about 1,0 ha. Thus, it can be concluded that the methodology presented has met the objectives proposed in the study and made it possible to attribute an economical meaning to the coefficient of water uniformity application.

Use of Medicinal Plants with Teratogenic and Abortive Effects by PregnantWomen in a City in Northeastern Brazil

Purpose The purpose of this study is to verify the use ofmedicinal plants by pregnant women treated at four Basic Health Units and at a public maternity facility in Brazil s northeast. Methods This is a cross-sectional, quantitative study, performed between February and April 2014. The subjects were 178 pregnant women, aged 18 to 42 years. To collect data, a structured questionnaire with dichotomous and multiple choice questions was used. To verify the correlation between the variables, Pearson s chi-square test was used. Results The study showed that 30.9% of the pregnant women used medicinal plants, and boldo was the most cited (35.4%). All the plants utilized, except lemongrass, have toxic effects in pregnancy, according to Resolution SES/RJ N° 1757. There was no statistically significant correlation between social class and use of medicinal plants. Conclusion The health of the study participants and their unborn children is at risk due to the inappropriate use of medicinal plants.

Occurrence of weed species in Jatropha curcas intercropping systems

This study aimed to evaluate the level of infestation by weed species in a consolidated Jatropha plantation, as a function of the plant species grown in interrows. The experiment was installed in 2006 at the district of Itahum, city of Dourados, state of Mato Grosso do Sul, Brazil, made possible through a partnership between Embrapa Western Agriculture and Paraiso Farm. Treatments consisted of (1) Jatropha in monocrop, with no plant at the interrrows; or the following plants cultivaded at the interrows of Jatropha: (2) Stylosanthes spp.; (3) Brachiaria ruziziensis; (4) Brachiaria ruziziensis + Stylosanthess pp.; (5) Brachiaria humidicola; (6) Panicum maximum cv. massai; (7) Cajanus cajan cv. anão; (8) Crotalaria spectabilis; (9) Crop rotation system 1 - (maize second crop -Crambe abyssinica - soybean - peanut); and (10) Crop rotation system 2 - (cowpea - radish - maize - cowpea) conducted for two years. Phytosociological characterization of weed species was accomplished in 2011 based on the Ecological Approach. Estimations of relative abundance, frequency, dominance and Importance Value Index were obtained. Areas were also characterized by the diversity coefficients of Simpson and modified Shannon-Weiner, and then grouped by cluster analysis. Areas with low soil coverage resulted in higher infestation levels; crop rotation in the interrows of Jatropha produced a significant reduction in weed infestation, but the lowest infestation levels were observed when grasses were grown intercropped with Jatropha. Lower diversity coefficients were associated with occurrence of the most troublesome weed species. In the first years after planting Jatropha, species of Brachiaria or a crop rotation involving species with high mulching ability and whose biomass exhibit a high C:N ratio, should be established in the interrows to avoid problems with weeds infestation.

SELECTION OF HERBICIDES TARGETING THE USE IN CROP SYSTEMS CULTIVATED WITH SHOWY CROTALARIA

ABSTRACT The increase in the area planted with Crotalaria spectabilishas occurred by several factors, highlighting the potential to reduce the nematodes, nitrogen fixation and the high production of biomass. By becoming a species sown as a crop, it is necessary to control the weeds that coexist with showy crotalaria. This change in the use of this crop creates the possibility of this specie becoming a weed. The aim of this study was to assess the potential use of herbicides applied in preemergence and postemergence of C.spectabilisfor different purposes (control of volunteer and selectivity plants). Three experiments were installed in a greenhouse (two with herbicides applied in preemergence - in soils with distinct textural categories; and one experiment with herbicides applied in postemergence). The results of the experiments with herbicides applied in preemergence showed that: amicarbazone, atrazine, diuron, metribuzin, prometryn, fomesafen and sulfentrazone showed effectiveness for control of C.spectabilis in clayey soil. Besides these, flumioxazin and isoxaflutole also showed potential to be used in the control of showy crotalaria in soils with loam texture. In relation to the postemergence herbicides, atrazine, diuron, prometryn, flumioxazin, fomesafen, lactofen, saflufenacil, amonio-glufosinate and glyphosate can be used aiming the chemical control of C.spectabilis. Herbicides chlorimuron-ethyl, diclosulan, imazethapyr, pyrithiobac-sodium, trifloxysulfuron-sodium, clomazone, pendimethalin, S-metolachlor and trifluralin applied in preemergence, and imazethapyr, pyrithiobac-sodium, flumiclorac, bentazon and clethodim applied in postemergence caused low levels of injury to C.spectabilis plants, making necessary the development of new searches to ensure the selectivity of these products.