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In this work, the susceptibility to benznidazole of two parental Trypanosoma cruzi strains, Colombian and Berenice-78, was compared to isolates obtained from dogs infected with these strains for several years. In order to evaluate the susceptibility to benznidazole two groups of mice were infected with one of five distinct populations isolated from dogs as well as the two parental strains of T. cruzi. The first group was treated with benznidazole during the acute phase and the second remained untreated controls. The animals were considered cured when parasitological and serological tests remained persistently negative. Mice infected with the Colombian strain and its isolates Colombian (A and B) did not cure after treatment. On the other hand, all animals infected with Berenice-78 were cured by benznidazole treatment. However, 100%, 50% and 70% of cure rates were observed in animals infected with the isolates Berenice-78 B, C and D, respectively. No significant differences were observed in serological profile of infected control groups, with all animals presenting high antibody levels. However, the ELISA test showed differences in serological patterns between mice inoculated with the different T. cruzi isolates and treated with benznidazole. This variability was dependent on the T. cruzi population used and seemed to be associated with the level of resistance to benznidazole.

Introduction: The consumption of red wine has historically been associated with a reduction in cardiovascular risk, with sometimes controversial scientific evidence., Method: A survey was carried out via whatsapp dated 09/01/22 to a cohort of doctors from the province of Malaga, asking about possible healthy red wine consumption habits, differentiating: never, 3-4 glasses per week, 5 -6 weekly drinks and one daily drink., Results: A total of 184 physicians answered, with a mean age of 35 years ± 11.1, of which 84 (45.6%) were women, distributed in different specialties, the majority being internal medicine with 52 (28.2%). The most frequently chosen option was D (59.2%), followed by A (21.2%), C (14.7%) and B (5%)., Conclusions: More than half of the doctors surveyed recommended zero consumption, and only 20% indicated that a daily drink could be healthy in non-drinkers., (Copyright © 2023 Sociedad Española de Arteriosclerosis. Publicado por Elsevier España, S.L.U. All rights reserved.)

Background: Activity assays for lipoprotein lipase (LPL) are not standardised for use in clinical settings., Objective: This study sought to define and validate a cut-off points based on a ROC curve for the diagnosis of patients with familial chylomicronemia syndrome (FCS). We also evaluated the role of LPL activity in a comprehensive FCS diagnostic workflow., Methods: A derivation cohort (including an FCS group (n = 9), a multifactorial chylomicronemia syndrome (MCS) group (n = 11)), and an external validation cohort (including an FCS group (n = 5), a MCS group (n = 23) and a normo-triglyceridemic (NTG) group (n = 14)), were studied. FCS patients were previously diagnosed by the presence of biallelic pathogenic genetic variants in the LPL and GPIHBP1 genes. LPL activity was also measured. Clinical and anthropometric data were recorded, and serum lipids and lipoproteins were measured. Sensitivity, specificity and cut-offs for LPL activity were obtained from a ROC curve and externally validated., Results: All post-heparin plasma LPL activity in the FCS patients were below 25.1 mU/mL, that was cut-off with best performance. There was no overlap in the LPL activity distributions between the FCS and MCS groups, conversely to the FCS and NTG groups., Conclusion: We conclude that, in addition to genetic testing, LPL activity in subjects with severe hypertriglyceridemia is a reliable criterium in the diagnosis of FCS when using a cut-off of 25.1 mU/mL (25% of the mean LPL activity in the validation MCS group). We do not recommend the NTG patient based cut-off values due to low sensitivity., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2023 National Lipid Association. Published by Elsevier Inc. All rights reserved.)

Repeated assessments in everyday life enables collecting ecologically valid data on dynamic, within-persons processes. These methods have widespread utility and application and have been extensively used for the study of stressors and stress responses. Enhanced conceptual sophistication of characterizing intraindividual stress responses in everyday life would help advance the field. This article provides a pragmatic overview of approaches, opportunities, and challenges when intensive ambulatory methods are applied to study everyday stress responses in "real time." We distinguish between three stress-response components (i.e., reactivity, recovery, and pileup) and focus on several fundamental questions: (a) What is the appropriate stress-free resting state (or "baseline") for an individual in everyday life? (b) How does one index the magnitude of the initial response to a stressor (reactivity)? (c) Following a stressor, how can recovery be identified (e.g., when the stress response has completed)? and (d) Because stressors may not occur in isolation, how can one capture the temporal clustering of stressors and/or stress responses (pileup)? We also present initial ideas on applying this approach to intervention research. Although we focus on stress responses, these issues may inform many other dynamic intraindividual constructs and behaviors (e.g., physical activity, physiological processes, other subjective states) captured in ambulatory assessment.

Purpose: Our goal was to analyze the outcome of infection and response to benznidazole (BZ) treatment in mice intragastrically inoculated with trypomastigotes forms of Trypanosoma cruzi from different origins., Methods: Twenty-four Swiss mice were divided in two groups and inoculated, by gavage, with 1 × 10 4 blood trypomastigotes (BT) or insect-derived metacyclic trypomastigotes (IT) of AM14 strain (T. cruzi IV). Half of the animals of each group were treated with BZ (TBZ), from 10 to 30th days after the inoculation, and the other constituted the untreated control groups (NT). After the etiological treatment, all mice were immunosuppressed with cyclophosphamide for three weeks. Parasitological and molecular parameters, infectivity, cumulative mortality, and reactivation post-immunosuppression rates were obtained., Results: Animals inoculated with BT showed lower pre-patent period and early day of the maximum parasitemia, as well as a higher maximum peak of parasitemia than the IT animals. However, both, BT and IT animals, did not respond to BZ treatment (0.0% of cure)., Conclusion: We conclude that the infective form influences in the outcome of infection, but not the response to the etiological treatment in mice intragastrically infected with the T. cruzi IV strain studied., (© 2021. Witold Stefański Institute of Parasitology, Polish Academy of Sciences.)

Background: Trypanosoma cruzi is the etiological agent of Chagas disease (CD) or American trypanosomiasis, an important public health problem in Latin America. Benznidazole (BZ), a drug available for its treatment, has limited efficacy and significant side effects. Essential oils (EOs) have demonstrated trypanocidal activity and may constitute a therapeutic alternative. Our aim was to evaluate the efficacy of the EOs of clove (CEO - Syzygium aromaticum) and ginger (GEO - Zingiber officinale), administered alone and in combination with BZ, in Swiss mice infected with T. cruzi., Methods: The animals were inoculated with 10,000 blood trypomastigotes of the Y strain of T. cruzi II by gavage and divided into four groups (n = 12 to 15): 1) untreated control (NT); 2) treated with BZ; 3) treated with CEO or GEO; and 4) treated with BZ + CEO or GEO. The treatments consisted of oral administration of 100 mg/kg/day, from the 5th day after parasite inoculation, for 20 consecutive days. All groups were submitted to fresh blood examination (FBE), blood culture (BC), conventional PCR (cPCR) and real-time PCR (qPCR), before and after immunosuppression with cyclophosphamide., Results: Clove and ginger EOs, administered alone and in combination with BZ, promoted suppression of parasitemia (p < 0.0001), except for the animals treated with CEO alone, which presented a parasitemia curve similar to NT animals. However, there was a decrease in the BC positivity rate (p < 0.05) and parasite load (< 0.0001) in this group. Treatment with GEO alone, on the other hand, besides promoting a decrease in the BC positivity rate (p < 0.05) and parasite load (p < 0.01), this EO also resulted in a decrease in mortality rate (p < 0.05) of treated mice., Conclusions: Decreased parasite load, as detected by qPCR, was observed in all treatment groups (BZ, CEO, GEO and BZ + EOs), demonstrating benefits even in the absence of parasitological cure, thus opening perspectives for further studies.

The whole-body insulin sensitivity index (WBISI) is a widely used surrogate of insulin sensitivity estimated from glucose and insulin concentrations during an oral glucose tolerance test (OGTT). The original WBISI uses five timepoints but reduced sampling models have been proposed. These reduced models have not been applied to the paediatric population. The purpose of this study is to compare cross-sectional agreement and changes in response to lifestyle intervention between the original WBISI and the WBISI 120 using fasting and 2-hour glucose and insulin concentrations from OGTT among Latino adolescents with obesity. We also examined the cost-differential between the two measures. Secondary analyses were conducted with data from Latino adolescents (14-16 years) with obesity (BMI ≥ 95th percentile) who were recruited for a randomized controlled lifestyle intervention trial. Baseline data in youth (n = 148) who underwent a 2-hour OGTT was used to analyse agreement between WBISI and WBISI 120 . Data from 65 participants who completed a 12-week lifestyle intervention were used to assess changes and the rate of change between WBISI and WBISI 120 . Research costs to determine both measures were compared. WBISI 120 showed good cross-sectional agreement (ICC agreement = 0.88) with the full WBISI. Following intervention, WBISI 120 increased 62.5% (M ± SD, 1.6 ± 1.2 to 2.6 ± 1.7, P < .001) while WBISI increased by 25.0% (1.6 ± 1.0 to 2.0 ± 1.0, P < .001) but the rate of change for WBISI and WBISI 120 was not significantly different (P = .11). WBISI 120 costs ~70% less than WBISI. WBISI 120 may offer a cost-effective surrogate estimate of insulin sensitivity in Latino youth with obesity., (© 2020 World Obesity Federation.)

The objective of this investigation was to evaluate the activity of the suprahyoid musculature during swallowing and to correlate the findings with the degree of megaesophagus, oral and pharyngeal videofluoroscopy and esophageal manometry in patients with achalasia caused by Chagas' disease. Twenty-nine patients with positive serology for Trypanosoma cruzi and dysphagia (Chagas' disease group) and 29 individuals matched by sex and age (control group) participated in the study. Surface electromyography of the suprahyoid musculature and videofluoroscopy during swallowing of paste and liquid consistencies were performed. Canonical correlation analysis of the MANOVA test results showed that the Chagas' disease group had lower electromyographic activity when compared with controls. Overlapping circles of radiological findings were found for megaesophagus. The Spearman test showed a positive correlation between the electromyographic activity in the maximum voluntary isometric contraction and the time of pharyngeal transit for both liquid (p = 0.014) and paste (p = 0.047). The logistic regression test showed no association between electromyographic activity of the suprahyoid muscles and esophageal manometry results (p > 0.05). In conclusion, individuals with chagasic megaesophagus have reduced electromyographic activity of the suprahyoid muscles during swallowing, in addition to a greater recruitment of the suprahyoid musculature with increased pharyngeal transit time.

Background: Trypanosoma cruzi, a hemoflagellate protozoan parasite and the etiological agent of Chagas disease (CD), exhibits great genetic and biological diversity. Infected individuals may present clinical manifestations with different levels of severity. Several hypotheses have been proposed to attempt to correlate the diversity of clinical signs and symptoms to the genetic variability of T. cruzi. This work aimed to investigate the differential expression of proteins from two distinct genetic groups of T. cruzi (discrete typing units TcI and TcII), isolated from chronically infected individuals displaying the cardiac form of CD. For this purpose, epimastigote forms of the two isolates were cultured in vitro and the cells recovered for protein extraction. Comparative two-dimensional (2D) gel electrophoreses were performed and differentially expressed spots selected for identification by mass spectrometry, followed by database searching and protein categorization., Results: The 2D electrophoretic profiles revealed the complex composition of the T. cruzi extracted proteome. Protein spots were distributed along the entire pH and molecular mass ranges attesting for the integrity of the protein preparations. In total, 46 differentially expressed proteins were identified present in 40 distinct spots found in the comparative gel analyses. Of these, 16 displayed upregulation in the gel from TcI-typed parasites and 24 appeared overexpressed in the gel from TcII-typed parasites. Functional characterization of differentially expressed proteins revealed major alterations associated with stress response, lipid and amino acid metabolism in parasites of the TcII isolate, whilst those proteins upregulated in the TcI sample were primarily linked to central metabolic pathways., Conclusions: The comparative 2D-gel electrophoresis allowed detection of major differences in protein expression between two T. cruzi isolates, belonging to the TcI and TcII genotypes. Our findings suggest that patients displaying the cardiac form of the disease harbor parasites capable of exhibiting distinct proteomic profiles. This should be of relevance to disease prognosis and treatment.

Objective: Sit-stand workstations may result in significant reductions in workplace sitting. However, few studies have examined long-term maintenance under real-world conditions. The purpose of this study was to evaluate workplace sitting time, cardio-metabolic biomarkers, and work productivity during a workplace re-design which included the installation of sit-stand workstations., Design: Natural experiment with appropriately matched comparison., Methods: Office workers from distinct worksites in the same unit were recruited (Intervention, n=24; Comparison, n=12). Intervention arm participants received a sit-stand workstation and 4 months of sitting-specific motivational support. The comparison arm received 4 months of ergonomic focused motivational support. Time spent in sitting, standing, and other physical activity were measured by activPAL3c for a week. Cardio-metabolic biomarkers and work productivity were also measured. Assessments occurred at baseline, 4 months, and 18 months., Results: At 4 months, work sitting time was reduced by 56.7±89.1min/8h workday (d=-0.64), relative to comparison. Standing time (37.4±69.2min/8h workday; d=0.54) and sit-to-stand transitions (3.3±0.4min/8h workday, d=0.44) were also improved relative to comparison. At 18 months, work sitting time reductions (52.6±68.3min/8h workday; d=-0.77) and standing time improvements (17.7±54.8min/8h workday, d=0.32) were maintained in the intervention group relative to comparison. Cardio-metabolic and work productivity changes were mixed; however, strongest effects favoring the intervention group were observed at 18 months., Conclusions: Sit-stand workstations, accompanied with behavioral support, were effective in reducing workplace and overall daily sitting and increasing standing time in a real-world setting. The effect appears to have been sustained for 18 months, with mixed results in cardio-metabolic and productivity outcomes., (Copyright © 2017 Sports Medicine Australia. Published by Elsevier Ltd. All rights reserved.)

Background: Although current technological advancements have allowed for objective measurements of sedentary behavior via accelerometers, these devices do not provide the contextual information needed to identify targets for behavioral interventions and generate public health guidelines to reduce sedentary behavior. Thus, self-reports still remain an important method of measurement for physical activity and sedentary behaviors., Objective: This study evaluated the reliability, validity, and sensitivity to change of a smartphone app in assessing sitting, light-intensity physical activity (LPA), and moderate-vigorous physical activity (MVPA)., Methods: Adults (N=28; 49.0 years old, standard deviation [SD] 8.9; 85% men; 73% Caucasian; body mass index=35.0, SD 8.3 kg/m2) reported their sitting, LPA, and MVPA over an 11-week behavioral intervention. During three separate 7-day periods, participants wore the activPAL3c accelerometer/inclinometer as a criterion measure. Intraclass correlation (ICC; 95% CI) and bias estimates (mean difference [δ] and root of mean square error [RMSE]) were used to compare app-based reported behaviors to measured sitting time (lying/seated position), LPA (standing or stepping at <100 steps/minute), and MVPA (stepping at >100 steps/minute)., Results: Test-retest results suggested moderate agreement with the criterion for sedentary time, LPA, and MVPA (ICC=0.65 [0.43-0.82], 0.67 [0.44-0.83] and 0.69 [0.48-0.84], respectively). The agreement between the two measures was poor (ICC=0.05-0.40). The app underestimated sedentary time (δ=-45.9 [-67.6, -24.2] minutes/day, RMSE=201.6) and overestimated LPA and MVPA (δ=18.8 [-1.30 to 38.9] minutes/day, RMSE=183; and δ=29.3 [25.3 to 33.2] minutes/day, RMSE=71.6, respectively). The app underestimated change in time spent during LPA and MVPA but overestimated change in sedentary time. Both measures showed similar directions in changed scores on sedentary time and LPA., Conclusions: Despite its inaccuracy, the app may be useful as a self-monitoring tool in the context of a behavioral intervention. Future research may help to clarify reasons for under- or over-reporting of behaviors., (©Meynard John Toledo, Eric Hekler, Kevin Hollingshead, Dana Epstein, Matthew Buman. Originally published in JMIR Mhealth and Uhealth (http://mhealth.jmir.org), 09.08.2017.)

Trypanosoma cruzi is the etiologic agent of American trypanosomiasis has broad biological and genetic diversity. Remaining to be studied are polymorphisms of the blood forms and metacyclogenesis of different T. cruzi discrete typing units (DTUs). Our goal was to evaluate the relationship between T. cruzi DTUs, the morphology of blood trypomastigotes, and in vitro metacyclogenesis. T. cruzi strains that pertained to DTUs TcI, TcII, and TcIV from different Brazilian states were used. Parameters that were related to the morphology of eight strains were assessed in thin blood smears that were obtained from mice that were inoculated with blood or culture forms, depending on strain. The metacyclogenesis of 12 strains was measured using smears with Liver Infusion Tryptose culture medium and M16 culture medium (which is poor in nutrients and has a low pH) at the exponential phase of growth, both stained with Giemsa. The morphological pattern of TcII strains was consistent with broad forms of the parasite. In TcIV strains, slender forms predominated. The Y strain (TcII) was morphologically more similar to TcIV. Significant differences in polymorphisms were observed between DTUs. Metacyclogenesis parameters, although displaying large standard deviations, differed between the DTUs, with the following descending rank order: TcII > TcI > TcIV. The mean numbers of metacyclic trypomastigotes for TcII were significantly higher than the other DTUs. Although the DTUs presented overlapping characteristics, the general pattern was that different DTUs exhibited significantly different morphologies and metacyclogenesis, suggesting that the genetic diversity of T. cruzi could be related to parameters that are associated with the evolution of infection in mammalian hosts and its ability to disperse in nature., (Copyright © 2017 Elsevier Inc. All rights reserved.)

A new epidemiological view of American trypanosomiasis or Chagas disease has been formulated in recent decades. Oral transmission of the etiological agent of Chagas disease, Trypanosoma cruzi, has been the most common form of transmission. The T. cruzi discrete typing units TcI and TcIV have been involved in tens outbreaks of acute cases of Chagas disease in the Brazilian Amazon region. We investigated the intensity of infection in mice that were orally inoculated (OR group) with four strains of TcIV that were isolated from two outbreaks of acute Chagas disease that was orally acquired in the state of Amazonas, Brazil. We compared the OR group with mice that were intraperitoneally inoculated (IP group). Blood samples were analyzed by fresh blood examination, hemoculture, and conventional and qualitative real-time polymerase chain reaction (PCR). Samples of different tissues were analyzed by quantitative real-time PCR. The OR group exhibited a higher maximum peak of parasitemia, greater rates of positivity, and higher parasite loads in different tissues during acute infection compared with the IP group, indicating a greater intensity of orally acquired infection. Mice that were orally inoculated with TcIV strains that were obtained from two outbreaks of orally acquired Chagas disease in Amazonas, Brazil, exhibited a more intense course of infection compared with intraperitoneally inoculated mice, reflected by higher levels of parasitemia and parasite loads., (Copyright © 2016. Published by Elsevier B.V.)

Background: American workers spend 70-80% of their time at work being sedentary. Traditional approaches to increase moderate-vigorous physical activity (MVPA) may be perceived to be harmful to productivity. Approaches that target reductions in sedentary behavior and/or increases in standing or light-intensity physical activity [LPA] may not interfere with productivity and may be more feasible to achieve through small changes accumulated throughout the workday METHODS/DESIGN: This group randomized trial (i.e., cluster randomized trial) will test the relative efficacy of two sedentary behavior focused interventions in 24 worksites across two states (N=720 workers). The MOVE+ intervention is a multilevel individual, social, environmental, and organizational intervention targeting increases in light-intensity physical activity in the workplace. The STAND+ intervention is the MOVE+ intervention with the addition of the installation and use of sit-stand workstations to reduce sedentary behavior and enhance light-intensity physical activity opportunities. Our primary outcome will be objectively-measured changes in sedentary behavior and light-intensity physical activity over 12months, with additional process measures at 3months and longer-term sustainability outcomes at 24months. Our secondary outcomes will be a clustered cardiometabolic risk score (comprised of fasting glucose, insulin, triglycerides, HDL-cholesterol, and blood pressure), workplace productivity, and job satisfaction DISCUSSION: This study will determine the efficacy of a multi-level workplace intervention (including the use of a sit-stand workstation) to reduce sedentary behavior and increase LPA and concomitant impact on cardiometabolic health, workplace productivity, and satisfaction., Trial Registration: ClinicalTrials.gov Identifier: NCT02566317 (date of registration: 10/1/2015)., (Copyright © 2016 Elsevier Inc. All rights reserved.)

The spatial distribution of enteroparasitosis in an indigenous village from Paraná was evaluated to identify areas of risk for these infections. A cross-sectional study (from November 2010 to June 2011) was performed using Three Faecal Test(®) and Kato & Katz method and a questionnaire on housing and hygiene conditions was administered. Local geostatistical analyses were performed to determine the spatial distribution of intestinal parasitic infections. The overall prevalence of enteroparasites was 67.2 % (457/680), and the most prevalent taxa were Ascaris lumbricoides (48.8 %) and Trichuris trichiura (44.7 %). The prevalence of heavy infection by soil-transmitted helminths was 3.6 % and the families lived in houses with an average of 5.1 residents and < 2 bedrooms per household. The average number of species per individual present spatial heterogeneity with the highest values (≥0.8) in areas with high clustering of residences. The visualization of the spatial distribution of intestinal parasites in this indigenous village is an important contribution to determining health risk areas and planning decisions and services.

Energy expenditure (EE) estimation from accelerometer-based wearable sensors is important to generate accurate assessment of physical activity (PA) in individuals. Approaches hitherto have mainly focused on using accelerometer data and features extracted from these data to learn a regression model to predict EE directly. In this paper, we propose a novel framework for EE estimation based on statistical estimation theory. Given a test sequence of accelerometer data, the probability distribution on the PA classes is estimated by a classifier and these predictions are used to estimate EE. Experimental evaluation, performed on a large dataset of 152 subjects and 12 activity classes, demonstrates that EE can be estimated accurately using our framework.

Background: The study evaluated qualitative PCR, primers 121-122 as a tool to follow up evolution parasite load of Trypanosoma cruzi ., Methods: The study was conducted at the State University of Maringa, in 2015. Step 1, dilutions 1/10 were performed from T. cruzi -Y strain to obtain preparations of 50,000-0.05 parasites/mL from which DNA were extracted, quantified, and amplified. Step 2, the extracted DNA in the dilutions 5-0.05 parasites/mL was re-diluted 1/10, 1/100, 1/1000, quantified, and amplified. Polyacrylamide gels were photographed and thicknesses of the 330 bp kDNA fragments were measured., Results: Step 1, in the dilutions 50,000-50 parasites/mL kDNA fragments had same thickness and, dilutions 5-0.05 parasites/mL showed progressive decrease in thicknesses and staining intensity of the 330 bp fragments. Step 2, demonstrated that dilutions of five (re-dilutions 1/10 and 1/100) and 0.5 (1/10) parasites/mL produced similar thicknesses of the 330 bp fragments obtained in Step 1. However, very dilute DNA samples make difficult to reproduce the fragments thicknesses., Conclusion: PCR, despite its limitations, was able to detect progressive decrease in thicknesses/staining intensity of kDNA fragments in the dilutions 5-0.05 parasites/mL. Hence, has the potential to be used to follow-up evolution of parasite load, not by quantifying the number of parasites, but by dynamic evolution of the fragments thicknesses during etiological treatment.

The prevalence and distribution of soil and water samples contaminated with enteroparasites of humans and animals with zoonotic potential (EHAZP) in Apucaraninha Indigenous Land (AIL), southern Brazil, was evaluated. An environmental survey was conducted to evaluate the presence of parasitic forms in peridomiciliary soil and associated variables. Soil samples were collected from 40/293 domiciles (10 domiciles per season), from November 2010 to June 2011, and evaluated by modified methods of Faust et al. and Lutz. Analyses of water from seven consumption sites were also performed. The overall prevalence of soil samples contaminated by EHAZP was 23.8 %. The most prevalent parasitic forms were cyst of Entamoeba spp. and eggs of Ascaris spp. The highest prevalence of contaminated soil samples was observed in winter (31 %). The probability map obtained with geostatistical analyses showed an average of 47 % soil contamination at a distance of approximately 140 m. The parasitological analysis of water did not detect Giardia spp. or Cryptosporidium spp. and showed that all collection points were within the standards of the Brazilian law. However, the microbiological analysis showed the presence of Escherichia coli in 6/7 sampled points. Despite the low level of contamination by EHAZP in peridomiciliar soil and the absence of pathogenic protozoa in water, the AIL soil and water (due to the presence of fecal coliforms) are potential sources of infection for the population, indicating the need for improvements in sanitation and water treatment, in addition periodic treatment of the population with antiparasitic.

The specific detection and genetic typing of trypanosomes that infect humans, mammalian reservoirs, and vectors is crucial for diagnosis and epidemiology. We utilized a PCR-RFLP assay that targeted subunit II of cytochrome oxidase and 24Sα-rDNA to simultaneously detect and discriminate six Trypanosoma cruzi discrete typing units (DTUs) and two genetic groups of Trypanosoma rangeli (KP1+/KP1-) in intestinal contents of experimentally infected Rhodnius prolixus. The PCR assays showed that in 23 of 29 (79.4%) mixed infections with the six T. cruzi DTUs and mixed infections with individual DTUs and/or groups KP1+ and KP1-, both parasites were successfully detected. In six mixed infections that involved TcIII, the TcI, TcII, TcV, and TcVI DTUs predominated to the detriment of TcIII, indicating the selection of genetic groups. Interactions between different genetic groups and vectors may lead to genetic selection over TcIII. The elimination of this DTU by the immune system of the vector appears unlikely because TcIII was present in other mixed infections (TcIII/TcIV and TcIII/KP1+). Both molecular markers used in this study were sensitive and specific, demonstrating their usefulness in a wide geographical area where distinct genotypes of these two species are sympatric. Although the cellular and molecular mechanisms that are involved in parasite-vector interactions are still poorly understood, our results indicate a dynamic selection toward specific T. cruzi DTUs in R. prolixus during mixed genotype infections., (Copyright © 2016 Elsevier B.V. All rights reserved.)

American trypanosomiasis is an emerging zoonosis in the Brazilian Amazon. Studies on benznidazole (BZ) chemotherapy with Trypanosoma cruzi from this region have great relevance, given the different discrete typing units (DTUs) that infect humans in the Amazon and other regions of Brazil. We performed a parasitological, histopathological, and molecular analysis of mice inoculated with strains of T. cruzi I, II, and IV that were BZ-treated during the acute phase of infection. Groups of Swiss mice were inoculated; 13 received oral BZ, whereas the other 13 comprised the untreated controls. Unlike parasitemia, the infectivity and mortality did not vary among the DTUs. Trypanosoma cruzi DNA was detected in all tissues analyzed and the proportion of organs parasitized varied with the parasite DTU. The BZ treatment reduced the most parasitological parameters, tissue parasitism and the inflammatory processes at all infection stages and for all DTUs. However, the number of significant reductions varied according to the DTU and infection phase., (© The American Society of Tropical Medicine and Hygiene.)

Unlabelled: This study evaluates the effect of Trypanosoma cruzi biotherapy 17dH (BIOT) on mice of different ages, infected with the protozoa concerned., Method: Performing a blind, controlled, randomized by drawing experiment, 110 animals four or eight-week-old, Swiss, male mice were divided into infected control treated hydroalcoholic 7% (CI-4 = 34 or CI-8 = 21 animals) and infected control treated with biotherapy 17dH-0.2 mL/animal/20 consecutive days/oral regimen (BIOT-4 = 33 or BIOT-8 = 21 animals). Animals were inoculated intraperitoneally with 1400 trypomastigote, T. cruzi Y-strain. Parasitological, immunological and histopathologic parameters were evaluated statistically, using Statistica-8.0 and R 3.0.2 program to analysis of survival. The study was approved by the Ethics Committee for Animal Experimentation/UEM., Results: Four-week-old mice showed no statistical difference in parasitemia (P = 0.5718) between the treated and control group. Eight-week-old mice from the treated group had a higher parasite peak (P = 0.0424) and higher parasitemia (P < 0.005) than the control. To both groups of 4 and 8 weeks of age, treated or untreated, survival of mice was higher in the treated group than in the control, although it was not statistically significant (p-value = 0.32, 0.55 respectively). Four-week-old mice displayed a spleen section with a number of amastigote nests significantly higher in BIOT-4 than CI-4 (P = 0.01). In eight-week-old mice the number of amastigote nests (P < 0.001) and inflammatory foci (P < 0.06-10% significance) in the liver section were smaller in BIOT-8 than CI-8. Spleen giant cells were significantly higher in CI-8 than in BIOT-8 (P < 0.01). Eight-week-old animals treated with biotherapy showed higher parasitemia and lower tissue parasitism. Opposite pattern was observed in four-week-old animals., Conclusion: There is a difference of high diluted medication effect in four and eight-week-old mice. In the group of animals 8 weeks the immunomodulatory effect seems to have been higher. Hence, treatment with the medicine produced from T. cruzi modulates the inflammatory response with increased apoptosis and decreased serum levels of TGF-β., (Copyright © 2014 The Faculty of Homeopathy. Published by Elsevier Ltd. All rights reserved.)

The geographical heterogeneity of Chagas disease (ChD) is mainly caused by genetic variability of the etiological agent Trypanosoma cruzi. Our hypothesis was that the pathogenicity for mice may vary with the genetic lineage (or Discrete Typing Unit - DTU) of the parasite. To test this hypothesis, parasitological and histopathological evaluations were performed in mice inoculated with strains belonging to the DTU T. cruzi IV (TcIV) from the State of Amazonas (northern Brazil), or the DTU T. cruzi II (TcII) from the State of Paraná (southern Brazil). Groups of 10 Swiss mice were inoculated with eight strains of TcIV obtained from acute cases (7) from two outbreaks of orally acquired ChD, and from the triatomine Rhodnius robustus (1) from Amazonas; and three strains of TcII obtained from chronic patients in Paraná. We evaluated the pre-patent period, patent period, maximum peak of parasitemia, day of maximum peak of parasitemia, area under the parasitemia curve, inflammatory process, and tissue parasitism in the acute phase. TcIV was less virulent than TcII, and showed significantly (p < 0.005) lower parasitemia levels. Although the levels of tissue parasitism did not differ statistically, mice infected with TcIV displayed significantly (p < 0.001) fewer inflammatory processes than mice infected with TcII. This supported the working hypothesis, since TcIV from Amazonas was less pathogenic than TcII from Paraná; and agreed with the lower severity of human cases of ChD in the Amazon region., (Copyright © 2014 Elsevier Inc. All rights reserved.)

Background: The parasites Trypanosoma rangeli and Trypanosoma cruzi share vectors and hosts over a wide geographical area in Latin America. In this study, we propose a single molecular approach for simultaneous detection and typing of T. rangeli and T. cruzi., Methods: A restriction fragment length polymorphism analysis of the mitochondrial cytochrome oxidase II gene (COII-RFLP) using enzyme AluI and different amounts of DNA from the major genetic groups of T. rangeli and T. cruzi (KP1+/KP1- and DTU-I/DTU-II) was carried out. The same marker was tested on the other T. cruzi DTUs (DTU-III to DTU-VI) and on DNA extracted from gut contents of experimentally infected triatomines., Results: The COII PCR generates a ~400 bp fragment, which after digestion with AluI (COII-RFLP) can be used to distinguish T. rangeli from T. cruzi and simultaneously differentiate the major genetic groups of T. rangeli (KP1+ and KP1-) and T. cruzi (DTU-I and DTU-II). The COII-RFLP generated bands of ~120 bp and ~280 bp for KP1+, whereas for KP1- no amplicon cleavage was observed. For T. cruzi, digestion of COII revealed a ~300 bp band for DTU-I and a ~250 bp band for DTU-II. For DTU-III to DTU-VI, COII-RFLP generated bands ranging from ~310 to ~330 bp, but the differentiation of these DTUs was not as clear as the separation between DTU-I and DTU-II. After AluI digestion, a species-specific fragment of ~80 bp was observed for all DTUs of T. cruzi. No cross-amplification was observed for Leishmania spp., T. vivax or T. evansi., Conclusions: The COII-RFLP allowed simultaneous detection and typing of T. rangeli and T. cruzi strains according to their major genetic groups (KP1+/KP1- and DTU-I/DTU-II) in vitro and in vivo, providing a reliable and sensitive tool for epidemiological studies in areas where T. rangeli and T. cruzi coexist.

This study evaluated the influence of moderate physical exercise on the myenteric neurons in the colonic intestinal wall of mice that had been infected with Trypanosoma cruzi. Parasitology and immunological aspects of the mice were considered. Forty-day-old male Swiss mice were divided into four groups: Trained Infected (TI), Sedentary Infected (SI), Trained Control (TC), and Sedentary Control (SC). The TC and TI were subjected to a moderate physical exercise program on a treadmill for 8 weeks. Three days after finishing exercise, the TI and SI groups were inoculated with 1,300 blood trypomastigotes of the Y strain-T. cruzi. After 75 days of infection results were obtained. Kruskal-Wallis or Analyze of variance (Tukey post hoc test) at 5% level of significance was performed. Moderate physical exercise reduced both the parasite peak (day 8 of infection) and total parasitemia compared with the sedentary groups (P < 0.05). This activity also contributed to neuronal survival (P < 0.05). Exercise caused neuronal hypertrophy (P < 0.05) and an increase in the total thickness of the intestinal wall (P < 0.05). The TI group exhibited an increase in the number of intraepithelial lymphocytes (P > 0.05). In trained animals, the number of goblet cells was reduced compared with sedentary animals (P < 0.05). Physical exercise prevented the formation of inflammatory foci in the TI group (P < 0.05) and increased the synthesis of TNF-α (P < 0.05) and TGF-β (P > 0.05). The present results demonstrated the benefits of moderate physical exercise, and reaffirmed the possibility of that it may contribute to improving clinical treatment in Chagas' disease patients., (© 2013 The Authors. International Journal of Experimental Pathology © 2013 International Journal of Experimental Pathology.)

Oral infection has become the most important transmission mechanism of Chagas disease in Brazil. For this study, the development of Trypanosoma cruzi infection in mice, induced by the oral and intraperitoneal (IP) routes, was compared. Four groups of Swiss mice were used to evaluate the influence of parasite genetics, number of parasites, inoculation volume and developmental stages on the development of the orally induced infection: 1 - blood trypomastigotes (BT) via oral; 2 - BT via IP; 3 - culture metacyclic trypomastigotes (MT) via oral; and 4 - culture MT via IP. Animals inoculated orally showed levels of parasitemia, as well as infectivity and mortality rates, lower than animals inoculated via IP, regardless of DTU (discrete typing unit) and inoculum. Animals infected with TcII showed higher levels of these parameters than did animals infected with TcI. The larger volume of inoculum showed a greater capacity to cause an infection when administered via the oral route. BT infection was more virulent than culture MT infection for both routes (oral and IP). However, mice inoculated orally with BT showed lower levels than via IP, while mice inoculated orally with culture MT showed similar levels of infection to those inoculated via IP. Mice inoculated with culture MT showed more histopathological changes than those inoculated with BT, regardless of the inoculation route. These results indicate that this alternative experimental model is useful for evaluating infection by T. cruzi isolates with subpatent parasitemia and low virulence, such as those belonging to the TcI and TcIV DTUs, which are prevalent in outbreaks of orally transmitted Chagas disease., (Copyright © 2013 Elsevier Inc. All rights reserved.)

Background: In the Brazilian Amazon, clinical and epidemiological frameworks of Chagas disease are very dissimilar in relation to the endemic classical areas of transmission, possibly due to genetic and biological characteristics of the circulating Trypanosoma cruzi stocks. Twenty six T. cruzi stocks from Western Amazon Region attributed to the TcI and TcIV DTUs were comparatively studied in Swiss mice to test the hypothesis that T. cruzi clonal structure has a major impact on its biological and medical properties., Methodology/principal Findings: Seventeen parameters were assayed in mice infected with 14 T. cruzi strains belonging to DTU TcI and 11 strains typed as TcIV. In comparison with TcI, TcIV stocks promoted a significantly shorter pre-patent period (p<0.001), a longer patent period (p<0.001), higher values of mean daily parasitemia (p = 0.009) and maximum of parasitemia (p = 0.015), earlier days of maximum parasitemia (p<0.001) and mortality (p = 0.018), higher mortality rates in the acute phase (p = 0.047), higher infectivity rates (p = 0.002), higher positivity in the fresh blood examination (p<0.001), higher positivity in the ELISA at the early chronic phase (p = 0.022), and a higher positivity in the ELISA at the late chronic phase (p = 0.003). On the other hand TcI showed higher values of mortality rates in the early chronic phase (p = 0.014), higher frequency of mice with inflammatory process in any organ (p = 0.005), higher frequency of mice with tissue parasitism in any organ (p = 0.027) and a higher susceptibility to benznidazole (p = 0.002) than TcIV. Survival analysis showing the time elapsed from the day of inoculation to the beginning of the patent period was significantly shorter for TcIV strains and the death episodes triggered following the infection with TcI occurred significantly later in relation to TcIV. The notable exceptions come from positivity in the hemocultures and PCR, for which the results were similar., Conclusion/significance: T. cruzi stocks belonging to TcI and TcIV DTUs from Brazilian Amazon are divergent in terms of biological and medical properties in mice.

Objective: To assess the susceptibility of Trypanosoma cruzi strains from Amazon to benznidazole., Methods: We studied 23 strains of T. cruzi obtained from humans in the acute phase of Chagas disease, triatomines and marsupials in the state of Amazonas and from chronic patients and triatomines in the state of Paraná, Brazil. The strains were classified as TcI (6), TcII (4) and TcIV (13). For each strain, 20 Swiss mice were inoculated: 10 were treated orally with benznidazole 100 mg/kg/day (TBZ group) for 20 consecutive days and 10 comprised the untreated control group (NT). Fresh blood examination, haemoculture (HC), PCR, and ELISA were used to monitor the cure., Results: The overall cure rate was 60.5% (109/180 mice) and varied widely among strains. The strains were classified as resistant, partially resistant or susceptible to benznidazole, irrespective of discrete typing units (DTUs), geographical origin or host. However, the TcI strains from Amazonas were significantly (P = 0.028) more sensitive to benznidazole than the TcI strains from Paraná. The number of parasitological, molecular and serological parameters that were significantly reduced by benznidazole treatment also varied among the DTUs; the TBZ group of mice inoculated with TcIV strains showed more reductions (8/9) than those with TcI and TcII strains., Conclusions: Benznidazole resistance was observed among natural populations of the parasite in the Amazon, even in those never exposed to the drug., (© 2012 Blackwell Publishing Ltd.)

The biological behaviour of 23 Trypanosoma cruzi isolates in Swiss mice was compared. Nineteen isolates were obtained from patients in the acute phase of Chagas disease (13), sylvatic reservoir hosts (Didelphis marsupialis) (3), and triatomine bugs (Rhodnius robustus) (3) from four regions of the State of Amazonas (AM). Four isolates were obtained from chronic chagasic patients in the State of Paraná (PR): three autochthones, and one allochthone from the State of Minas Gerais. Only one isolate was unable to infect the mice. The AM and PR isolates showed the largest number of significant differences from each other. The former had lower mean values in the pre-patent (5.4 days) and patent (4.6 days) periods (PP), with the parasitaemia (Pmax) reaching a peak of 9.9×10(4) blood trypomastigotes (BT)/mL of blood by the 7th day following inoculation. The AM isolates also had higher positivity to fresh-blood examination (FBE) (84.1%) compared to haemoculture (HC) (58.7%) and polymerase chain reaction (PCR) (33.3%), in addition to higher mortality (2.9%). The PR isolates had higher values for PP (18.5 days) and Pmax (99.9×10(4)BT/mL) as well as higher positivity to FBE (87.2%), HC (100%), and PCR (83.3%). The correlations between the biological behaviour of the T. cruzi isolates and the clinical and epidemiological characteristics of Chagas disease are discussed., (Copyright © 2012 Elsevier Inc. All rights reserved.)

Introduction: The biological diversity of circulating Trypanosoma cruzi stocks in the Amazon region most likely plays an important role in the peculiar clinic-epidemiological features of Chagas disease in this area., Methods: Seven stocks of T. cruzi were recently isolated in the State of Amazonas, Brazil, from humans, wild mammals, and triatomines. They belonged to the TcI and Z3 genotypes and were biologically characterized in Swiss mice. Parasitological and histopathological parameters were determined., Results: Four stocks did not promote patent parasitemia in mice. Three stocks produced low parasitemia, long pre-patent periods, and a patent period of 1 day or oscillating parasitemia. Maximum parasitemia ranged from 1,400 to 2,800 trypomastigotes/0.1 mL blood. Mice inoculated with the T. cruzi stocks studied showed low positivity during fresh blood examinations, ranging from 0% to 28.6%. In hemoculture, positivity ranged from 0% to 100%. Heart tissue parasitism was observed in mice inoculated with stocks AM49 and AM61. Stock AM49 triggered a moderate inflammatory process in heart tissue. A mild inflammatory process was observed in heart tissue for stocks AM28, AM38, AM61, and AM69. An inflammatory process was frequently observed in skeletal muscle. Examinations of brain tissue revealed inflammatory foci and gliosis in mice inoculated with stock AM49., Conclusions: Biological and histopathological characterization allowed us to demonstrate the low infectivity and virulence of T. cruzi stocks isolated from the State of Amazonas.

Background: Chagas disease is an emergent tropical disease in the Brazilian Amazon Region, with an increasing number of cases in recent decades. In this region, the sylvatic cycle of Trypanosoma cruzi transmission, which constitutes a reservoir of parasites that might be associated with specific molecular, epidemiological and clinical traits, has been little explored. The objective of this work is to genetically characterize stocks of T. cruzi from human cases, triatomines and reservoir mammals in the State of Amazonas, in the Western Brazilian Amazon., Methodology/principal Findings: We analyzed 96 T. cruzi samples from four municipalities in distant locations of the State of Amazonas. Molecular characterization of isolated parasites from cultures in LIT medium or directly from vectors or whole human blood was performed by PCR of the non-transcribed spacer of the mini-exon and of the 24 S alfa ribosomal RNA gene, RFLP and sequencing of the mitochondrial cytochrome c oxidase subunit II (COII) gene, and by sequencing of the glucose-phosphate isomerase gene. The T. cruzi parasites from two outbreaks of acute disease were all typed as TcIV. One of the outbreaks was triggered by several haplotypes of the same DTU. TcIV also occurred in isolated cases and in Rhodnius robustus. Incongruence between mitochondrial and nuclear phylogenies is likely to be indicative of historical genetic exchange events resulting in mitochondrial introgression between TcIII and TcIV DTUs from Western Brazilian Amazon. TcI predominated among triatomines and was the unique DTU infecting marsupials., Conclusion/significance: DTU TcIV, rarely associated with human Chagas disease in other areas of the Amazon basin, is the major strain responsible for the human infections in the Western Brazilian Amazon, occurring in outbreaks as single or mixed infections by different haplotypes.

The aim of this study was to characterise Discrete Typing Units (DTUs) of 28 isolates of Trypanosoma cruzi from humans (15), triatomines (9), and opossums (4) in the state of Paraná, southern Brazil. For this purpose, we analysed the size polymorphism at the 3' end of the 24Sα ribosomal RNA gene (rRNA) and the restriction fragment length polymorphism (RFLP) of the partial 5' sequence of the mitochondrial Cytochrome Oxidase subunit II gene (COII). Band patterns of the isolates were compared with reference samples of T. cruzi I (Silvio X10 and Col 17G2), T. cruzi II (Esmeraldo and JG), T. cruzi III (222 and 231), T. cruzi IV (CAN III), T. cruzi V (SO3 cl5), and T. cruzi VI (CL Brener). Our results confirmed that rRNA analysis is of limited use for assessing T. cruzi DTUs. COII RFLP analysis was suitable for screening, but for one isolate it was necessary to determine the COII partial sequence to identify the DTU. Only one of the isolates from humans belonged to T. cruzi I; 13 isolates belonged to T. cruzi II and one to T. cruzi III. The four isolates from opossums and five isolates from triatomines were identified as T. cruzi I. Four isolates from triatomines showed patterns of both T. cruzi I and II, indicating mixed infections. This study contributes to the characterisation of the dynamics of T. cruzi populations in southern Brazil., (Copyright © 2011 Elsevier B.V. All rights reserved.)

Twelve strains of Trypanosoma cruzi isolated from wild reservoirs, triatomines, and chronic chagasic patients in the state of Paraná, southern Brazil, and classified as T. cruzi I and II, were used to test the correlation between genetic and biological diversity. The Phagocytic Index (PI) and nitric-oxide (NO) production in vitro were used as biological parameters. The PI of the T. cruzi I and II strains did not differ significantly, nor did the PI of the T. cruzi strains isolated from humans, triatomines, or wild reservoirs. There was a statistical difference in the inhibition of NO production between T. cruzi I and II and between parasites isolated from humans and the strains isolated from triatomines and wild reservoirs, but there was no correlation between genetics and biology when the strains were analyzed independently of the lineages or hosts from which the strains were isolated. There were significant correlations for Randomly Amplified Polymorphic Deoxyribonucleic acid (RAPD) and biological parameters for T. cruzi I and II, and for humans or wild reservoirs when the lineages or hosts were considered individually.

Define an experimental model by evaluating quantitative and morphometric changes in myenteric neurons of the colon of mice infected with Trypanosoma cruzi. Twenty-eight Swiss male mice were distributed into groups: control (CG, n=9) and inoculated with 100 (IG(100), n=9) and 1000 (IG(1000), n=10) blood trypomastigotes, Y strain-T. cruzi II. Parasitemia was evaluated from 3-25 days post inoculation (dpi) with parasites peak of 7.7 × 10(6) and 8.4 × 10(6) trypomastigotes/mL at 8(th) dpi (p>0.05) in IG(100) and IG(1000), respectively. Chronic phase of the infection was obtained with two doses of 100mg/Kg/weight and one dose of 250mg/Kg/weight of Benznidazole on 11, 16 and 18 dpi. Three animals from each group were euthanized at 18, 30 and 75 dpi. The colon was stained with Giemsa. The quantitative and morphometric analysis of neurons revealed that the infection caused a decrease of neuronal density on 30(th) dpi (p<0.05) and 75 dpi (p<0.05) in IG(100) and IG(1000). Infection caused death and neuronal hypertrophy in the 75(th) dpi in IG(100) and IG(1000) (p<0.05, p<0.01). The changes observed in myenteric neurons were directly related to the inoculate and the time of infection.

The goals of this study were to evaluate the effect of the Canova medication, a homeopathic immune-system modulator, on the evolution of infection induced by the Trypanosoma cruzi Y strain in mice. The animals were divided into five groups: (i) untreated infected controls (I), (ii) infected animals treated with benznidazole (Bz), (iii) infected animals treated with the Canova medication (CM), (iv) infected animals treated with benznidazole and the Canova medication (Bz+CM), and (v) uninfected controls that received only the vehicle (grain alcohol) (C). The parameters evaluated were: parasitemia, mortality, control of cure, and tissue parasitism analysis. Our results showed that the evolution of the experimental infection was modified by treatment with CM, and that daily and consecutive doses were harmful to the animals, causing death in 100% of the infected animals in a brief period. The analysis of parasitism performed on the organs on the 12th day postinfection showed that in infected animals treated with CM, the number of amastigote/nests in the spleen was significantly reduced, while in cardiac tissue, intestine, and liver the number was significantly increased compared with infected control animals. These results indicate that CM has a negative influence on the host-parasite relationship, modifying the tropism of the parasite for tissues, and increasing the parasitemia peak in this experimental model.

Objectives: To determine 1) the degree of soil contamination by parasites; 2) the prevalence of contaminated samples; 3) the overall number of parasitic forms, by distance between the collection site and the domicile; 4) the mean number of parasite forms per peri-household area; and 5) the variables associated with the total number of parasite forms., Methods: Soil samples were collected in the peridomicile of 18 (out of a total sample of 63) and 22 (out of 190) residences of the Faxinal and Ivaí indigenous territories (ITs), respectively, from March 2005 through October 2006, and evaluated by the methods of Baermann, modified Faust et al., and Lutz., Results: The overall prevalence of samples contaminated by enteroparasites of humans or of animals with zoonotic potential was 75.7% (84 out of 111) in Faxinal and 96.2% (127 out of 132) in Ivaí. The most prevalent parasites were Ascaris spp., Isospora spp., and Toxocara spp. Positive statistical associations were observed between the number of parasitic forms per domicile and the following variables: time of year (season), the presence of bathrooms in the domicile, and the presence of excrement on peridomiciliar soil., Conclusions: The high level of soil contamination in Faxinal and Ivaí 1) constitutes a potential source of parasitic infection and 2) suggests that the treatment of human and animal excreta continues to be inadequate.

Introduction: Chagas disease is an emerging and neglected problem in the Brazilian Amazon region., Methods: This study describes a series of acute autochthonous cases of Chagas disease that were attended at the Tropical Medicine Foundation of Amazonas, Manaus, between 1980 and 2006., Results: Twenty-nine cases were recorded: 19 (65.5%) were male and 10 (34.5%) cases were female. Fifteen (51.7%) were isolated cases and 14 (48.3%) were from outbreaks. The commonest signs and symptoms were fever, fatigue, headache, myalgia, chills, pallor, dyspnea and edema of the face and lower limbs. No deaths were recorded., Conclusions: The disease occurred frequently among young people. The parasitological methods showed high sensitivity.

Objective: To analyze the parasitological state of families in an indigenous community after institution of enteroparasite control measures., Methods: A longitudinal study was conducted between 2004 and 2006 among 447 individuals of the Kaingáng ethnic group, in the municipality of Cândido de Abreu, Southern Brazil. The enteroparasite control measures were: sanitation improvements in 2003, antiparasite treatments that were administered during the study period, and health education activities that were started in 2005. Parasitological health indicators were obtained through three coproparasitological surveys (in 2004, 2005 and 2006), in which 250, 147 and 126 stool samples, respectively, were collected. These were evaluated using the spontaneous sedimentation, centrifugation-flotation and Kato-Katz methods. Housing and hygiene conditions were assessed by means of a questionnaire applied to 69 (2004), 57 (2005) and 38 (2006) of the 90 families., Results: The overall prevalences of enteroparasites were 91.6% (2004), 94.6% (2005) and 87.3% (2006) and did not show any significant reduction. The prevalence of some species decreased, while the prevalence of others increased significantly. High-intensity infections due to geohelminths presented rates of less than 2% over the study period. The proportion of the interviewees who reported using a toilet increased from 38.8% to 71.1% (p< 0.005) and the proportion taking antiparasite agents increased from 70.2% to 100% (p= 0.001)., Conclusions: There were significant improvements in the parasitological health indicators, such as reductions in the prevalence of some species of enteroparasites and maintenance of a low parasite load, thus showing the importance of combining antiparasite treatment with sanitation improvements.

The objective of this study was to investigate the occurrence of cysts of Giardia spp. and oocysts of Cryptosporidium spp. in waters of the Ivaí Indigenous Land, Brazil. Samples of river and spring water and of treated water were filtered and analyzed by direct immunofluorescence (Merifluor kit, Meridian Bioscience, Cincinnati, Ohio). Of 21 samples, 7 from each locality, 3 (3/7, 42.8%) from a river were positive for Giardia (mean concentration 2.57 cysts/L), and 1 (1/7, 14.3%) was positive for Cryptosporidium (6 oocysts/L). From springs, 1 sample (1/7, 14.3%) was positive for Cryptosporidium (6 oocysts/L). One sample (1/7, 14.3%) from treated water was positive for both, with 4 oocysts/L and 2 cysts/L. Giardia was the more frequent protozoan present.

We report a case of accidental infection by Trypanosoma cruzi in a 42-year-old female patient who presented an inoculation chagoma. Laboratory confirmation was based on examination of fresh blood, Giemsa-stained blood smear, immunoenzyme test (EIA-IgG), indirect immunofluorescence (IIF-IgM, IgG) and polymerase chain reaction (PCR). Only the PCR gave a positive result, and the EIA test was inconclusive. Two treatments with benznidazole were necessary. PCR was the only technique that continued to give positive results for approximately two months (65 days, or 2.2 months) following the second treatment and negative results from 96 days (3.2 months) to 850 days (28.3 months). We concluded that the presence of an inoculation chagoma and use of PCR were important and decisive for diagnosis and follow-up of the case.

Exercise performed before infections has been linked to improvement of the immune response against infections. The purpose of this study was to evaluate the influence of preinfection moderate-intensity treadmill training on acute Trypanosoma cruzi infection in mice. Ninety-nine female BALB/c mice were divided into 4 groups, as follows: training + infection (T+I) (n = 41); no training + infection (NT+I) (n = 38); training + no infection (T+NI) (n = 10); and no training + no infection (NT+NI) (n = 10). The exercise program for trained groups was carried out on a motorized treadmill for 8 weeks. Infected groups were inoculated with the Y strain of T. cruzi. Infectivity, prepatent period, patent period, parasitemia peak, mortality, survival time, weight, food intake, tumor necrosis factor-alpha serum levels, and peritoneal macrophage hydrogen peroxide production were evaluated. We found that preinfection training induced statistically significant reductions in parasitemia peak (p < 0.03) and weight loss (p < 0.04). However, no statistically significant differences were found for the other parameters evaluated when trained and nontrained infected groups were compared. We conclude that preinfection aerobic training induces some improvement in the immune response to T. cruzi infection in female BALB/c mice.

This study analyzes the health of Kaingáng women in the Faxinal indigenous territory in Paraná State, Brazil, with regard to risk factors for breast cancer. A descriptive and exploratory investigation was undertaken that included 95.4% of females 20 years or over. Data showed that 92.3% of Kaingáng women reported breastfeeding for more than one year; 41.4% had more than 5 children; mean age at menarche was 13.4 years; 61.5% had never heard of breast cancer; breast self-examination was only performed by 2.9%; 81.7% reported no clinical breast examination by a physician; 99% had never had a mammogram. No cases of breast or non-breast cancer were reported. The study highlights protective factors against carcinogenesis and the absence of key risk factors like hormone replacement therapy for more than five years and alcohol consumption in most Kaingáng women.

We present the case of a young woman who went to the Emergency Departent with macroscopic and anaemic haematuria. We also confirm that, when faced with the possibility of finding an arteriovenous fistula, arteriography is the best choice for diagnosis, and the best option for treatment is selective embolization.

The capacity of the polymerase chain reaction (PCR) to detect the DNA of Trypanosoma cruzi was evaluated in 90 blood samples from BALB/c mice infected with T. cruzi cloned stocks of genotypes 19 and 20 (T. cruzi I) and 39 and 32 (T. cruzi II), and treated with benznidazole. The results from the fresh blood examination, hemoculture, and ELISA allowed to group the treated animals into: cured (TC), dissociated (DIS) and non-cured (NC). The PCR detected T. cruzi DNA in 50.9%, 58.3% and 100.0% of the samples from TC, DIS and NC mice, respectively. These DNA possibly derives from live T. cruzi or from recently lysed parasites, suggests that these animals are in fact not cured. The difference between the PCR results and results obtained using other techniques was statistically significant and independent of the parasite genotype. The PCR described has therefore potential to be used in cure control of treated patients.

Chagas disease, caused by the protozoan Trypanosoma cruzi, involves immunomediated processes. Canova (CA) is a homeopathic treatment indicated in the diseases in which the immune system is depressed. This study evaluated the Random Amplification of Polymorphic DNA (RAPD) profile of T. cruzi under the influence of CA and Benznidazole (BZ). Mice infected with the genetic lineage of T. cruzi II (Y strain) were divided into 4 groups: Infected animals treated with saline solution (control group); treated with CA; treated with BZ; treated with CA and BZ combined. Treatment was given at the 5th-25th days of infection (D5-25). The parasites were isolated by haemoculture in Liver Infusion Tryptose (LIT) medium: at D5 (before treatment), D13, 15 and 25 (during treatment) and D55 and 295 (after treatment). DNA was extracted from the mass of parasites. RAPD was done with the primers lambdagt11-F, M13F-40 and L15996, the amplified products were eletrophoresed through a 4% polyacrylamide gel. Data were analyzed by the coefficient of similarity using the DNA-POP program. 163 markers were identified, 5 of them monomorphic. CA did not act against the parasites when used alone. The RAPD profiles of parasites treated with BZ and CA+BZ were different from those in the control group and in the group treated with CA. The actions of the CA and BZ were different and the action of BZ was different from the action of CA+BZ. These data suggest that CA may interact with BZ. The differences in the RAPD profile of the Y strain of T. cruzi produced by BZ, CA+BZ and the natural course of the infection suggest selection/suppression of populations.

The aim of this work was to investigate the impact of dual infections with stocks of Trypanosoma cruzi major genotypes on benznidazole (BZ) treatment efficacy. For this purpose, T. cruzi stocks representative of the genetic T. cruzi lineages, displaying different susceptibilities to BZ, belonging to the major T. cruzi genotypes broadly dispersed in North and South America and important in Chagas' disease epidemiology were used. Therapeutic efficacy was observed in 27.8% of the animals treated. Following BZ susceptibility classification, significant differences were observed in dual infections on the major genotype level, demonstrating that combinations of genotypes 19+39 and genotypes 19+32 led to a shift in the expected BZ susceptibility profile toward the resistance pattern. Analysis on the T. cruzi stock level demonstrated that 9 out of 24 dual infections shifted the expected BZ susceptibility profile compared with the respective single infections, including shifts toward lower and higher BZ susceptibilities. Microsatellite identification was able to identify a mixture of T. cruzi stocks in 7.7% of the T. cruzi isolates from infected and untreated mice (6.9%) and infected and treated but not cured mice (9.0%), revealing in some mixtures of BZ-susceptible and -resistant stocks that the T. cruzi stock identified after BZ treatment was previously susceptible in single infections. Considering the clonal structure and evolution of T. cruzi, an unexpected result was the identification of parasite subpopulations with distinct microsatellite alleles in relation to the original stocks observed in 12.2% of the isolates. Taken together, the data suggest that mixed infections, already verified in nature, may have an important impact on the efficacy of chemotherapy.

In this paper, the infectivity for mice of Trypanosoma cruzi I and II strains isolated from sylvatic animals, triatomines, and humans is determined using fresh blood examination, hemoculture, culture of macerated organs, and polymerase chain reaction (PCR). Six strains were considered to have low infectivity (9.1-18.2%), five medium (27.3-45.4%), and one high (100.0%). Infectivity of T. cruzi strains isolated from sylvatic animals was significantly higher than that of strains isolated from humans and triatomines (p=0.0141). No significant difference was observed between the infectivity of T. cruzi I and II strains. The parasite was detected by fresh blood examination in one strain, by hemoculture and culture of macerated organs in four strains, and by PCR in all strains. We conclude that the infectivity is related to the host from which the strains were isolated, but the infectivity is not related to the genetic group of the parasite. We also conclude that the majority of the strains studied have low and medium infectivity for mice, and that PCR is an important tool to detect T. cruzi in strains with this biological characteristic.

Herein, we have analyzed major biological properties following dual-clone Trypanosoma cruzi infections in BALB/c mice. Eight T. cruzi clonal stocks, two of each principal genotype, including genotype 19 and 20 (T. cruzi I), hybrid genotype 39 (T. cruzi) and 32 (T. cruzi II) were combined into 24 different dual-clone infections. Special attention was given to characterize biological parameters assayed including: prepatent period, patent period, maximum of parasitemia, day of maximum parasitemia, area under the parasitemia curve, infectivity, mortality, and hemoculture positivity. Our findings clearly demonstrated that features resultant of dual-clone infections of T. cruzi clonal stocks did not display either the characteristics of the corresponding monoclonal infections or the theoretical mixture based on the respective monoclonal infections. Significant changes in the expected values were observed in 4.2-79.2% of the mixtures considering the eight biological parameters studied. A lower frequency of significant differences was found for mixtures composed by phylogenetically distant clonal stocks. Altogether, our data support our hypothesis that mixed T. cruzi infections have a great impact on the biological properties of the parasite in the host and re-emphasizes the importance of considering the possible occurrence of natural mixed infections in humans and their consequences on the biological aspects of ongoing Chagas' disease.

The polymerase chain reaction showed high sensitivity for detecting Trypanosoma cruzi in the blood of mice, independent of clonal genotype (19, 20-T. cruzi I; 32, 39-T. cruzi II) or phase of the infection (acute or chronic).

This paper presents the genetic characterization of Trypanosoma cruzi strains isolated from chronic chagasic patients, triatomines, and sylvatic reservoirs from Paraná state, Southern Brazil, using the RAPD and SSR-PCR techniques. It has shown the presence of both phylogenetic groups of T. cruzi (I and II), describing for the first time the existence of T. cruzi II in Paraná state.