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Background: Visceral leishmaniasis (VL) in Brazil is a neglected, vector-borne, tropical parasitic disease that is responsible for several thousand human deaths every year. The transmission route involves sand flies becoming infected after feeding on infected reservoir host, mainly dogs, and then transmitting the Leishmania infantum parasites while feeding on humans. A major component of the VL control effort is the identification and euthanasia of infected dogs to remove them as a source of infection. A rapid, non-invasive, point-of-care device able to differentiate between the odours of infected and uninfected dogs may contribute towards the accurate diagnosis of canine VL. Methodology/Principal findings: We analysed the headspace volatile chemicals from the hair of two groups of dogs collected in 2017 and 2018 using a bench-top eNose volatile organic chemical analyser. The dogs were categorised as infected or uninfected by PCR analysis of blood samples taken by venepuncture and the number of parasites per ml of blood was calculated for each dog by qPCR analysis. We demonstrated using a robust clustering analysis that the eNose data could be discriminated into infected and uninfected categories with specificity >94% and sensitivity >97%. The eNose device and data analysis were sufficiently sensitive to be able to identify infected dogs even when the Leishmania population in the circulating blood was very low. Conclusions/Significance: The study illustrates the potential of the eNose to rapidly and accurately identify dogs infected with Le. infantum. Future improvements to eNose analyser sensor sensitivity, sampling methodology and portability suggest that this approach could significantly improve the diagnosis of VL infected dogs in Brazil with additional potential for effective diagnosis of VL in humans as well as for the diagnosis of other parasitic diseases. [ABSTRACT FROM AUTHOR]

Background: Dengue is a public health problem, and noncompliance with World Health Organization (WHO) recommendations for blood transfusion components is frequently reported. Moreover, economic impact studies of the WHO recommendations on the use of blood transfusion are scarce. Methods: We compared the cost and hospitalization time in a prospective observational study, by following hospitalised patients and analysing their medical records from 2010 and March 2016 to December 2017. We divided the patients into two groups: transfused (with or without WHO criteria for transfusion) and not transfused (with or without WHO criteria for transfusion). Generalised linear modelling was performed to identify the variable that could increase the costs and hospital stay. Results: Among 323 patients, 52 were transfused, of whom 52% without criteria (n = 27), and 271 were not transfused, of which 4.4% (n = 12) with criteria. Hospitalisation costs were 41% higher in the transfused group without criteria than in those with criteria (median US$ 674.3 vs US$ 478 p = 0.293). Patients who were not transfused but met the WHO criteria for transfusion (n = 12) had longer mean hospitalisation time than did those who were not transfused (3.8±3.4 days versus 3.6±3.1 days; p = 0.022). The GLM analysis using hospital stay and costs as the dependent variable explained approximately 33.4% (R2 = 0.334) of the hospitalisation time and 79.3% (R2 = 0.793) of costs. Receiving a transfusion increased the hospitalization time by 1.29 days (p = 0.0007; IRR = 1.29), and the costs were 5.1 times higher than those without receiving blood components (IRR = 5.1; p< 0.001; median US$ 504.4 vs US$ 170.7). In contrast, patients who were transfused according to WHO criteria had a reduction in costs of approximately 96% (IRR = 0.044; p<0.001; β = -3.12) compared to that for those who were not transfused according to WHO criteria (without criteria). Conclusion: Transfusion without following WHO recommendations increased the time and cost of hospitalisation. [ABSTRACT FROM AUTHOR]

Background: Visceral Leishmaniasis in humans presents with fever, anemia, and splenomegaly and can be lethal if not treated. Nevertheless, the majority of Leishmania infantum-infected individuals does not manifest symptoms and remain so provided they are not immunosuppressed. In this work, the performance of different tests was evaluated to detect asymptomatic individuals who were living in Teresina, Piauí state, Brazil, an endemic area for VL. Methodology: L. infantum-specific antibodies were detected by ELISA and two different rapid immunochromatographic (IC) diagnostic tests, Kalazar Detect and OnSite, and parasitic loads were detected by real time PCR [qPCR]. Additionally, we measured levels of the biomarkers monokine induced by IFN-γ (MIG) and IFN-γ-induced protein 10 (IP-10) before and after stimulation of whole blood with soluble Leishmania antigen [SLA]. Principal findings: Kalazar Detect and OnSite detected, respectively, 76% and 64% of patients presenting with active Visceral Leishmaniasis; 50% and 57% of patients remained positive in these tests, respectively, after treatment. Of the healthy participants in the study who were living in the endemic area, only 1.7% were positive with both of the IC tests. On the other hand, reactivity in ELISA tests revealed that 13% of these individuals presented asymptomatic infections; among VL patients, 84% presenting with active disease were reactive in ELISA, and after treatment, 55.5% were seropositive. L. infantum DNA was present in the blood of 37.9% of infected individuals living in the endemic area, while IP-10 and MIG biomarkers were detected in 26.7% of them. The greatest concordance of positivity occurred between ELISA and qPCR. Conclusion: The association of different techniques can detect asymptomatic infections, however, more research is necessary to develop ideal biomarkers that are simple to use in the clinic and in field studies in areas endemic for Visceral Leishmaniasis. [ABSTRACT FROM AUTHOR]

Detection and sequencing of chikungunya virus (CHIKV) genome was performed using a combination of a modified reverse transcription loop-mediated isothermal amplification (RT-LAMP) method and a MinION sequencer. We developed the protocol for drying all the reagents for the RT-LAMP in a single reaction tube. Using this system, the CHIKV genome was effectively amplified under isothermal conditions, and used as a template for MinION sequencing with a laptop computer. Our in-house RT-LAMP method and MinION sequencing system were also validated with RNAs and serum samples from recent outbreaks of CHIKV patients in Brazil. The obtained sequence data confirmed the CHIKV outbreaks and identified the genotype. In summary, our established inexpensive on-site genome detection and sequencing system is applicable for both diagnosis of CHIKV infected patients and genotyping of the CHIKV virus in future outbreak in remote areas. [ABSTRACT FROM AUTHOR]

Background: Phlebotomine sand flies are vectors for several pathogens, with Leishmania being the most important. In Brazil, the main aetiological agent of American cutaneous leishmaniasis (ACL) is Leishmania (Viannia) braziliensis, and Nyssomyia neivai is one of its main vectors in São Paulo state and other areas of South America. Similar to other haematophagous insects, sand flies use volatile compounds called kairomones to locate their hosts for blood meals. A possible increase in the attractiveness of hosts infected with Leishmania infantum to their vectors has been demonstrated. In the present study, we aimed to investigate whether L. braziliensis-infected hosts present higher attractiveness to Ny. neivai and to identify differences in the volatile compounds released by infected and uninfected mice. Results: Behavioural experiments in which sand fly females directly fed on infected or uninfected mice showed no significant differences in the attractiveness of the mice or the blood volume ingested. Y-tube olfactometer bioassays also revealed no significant differences in the attractiveness of these hosts to Ny. neivai. No differences were observed in the profiles of the volatile compounds released by the two groups of mice. However, PCA and cluster analysis were able to classify the 31 identified compounds into three clusters according to their abundances. This classification showed a possible role for individual variation in the absence of differences in volatile profiles and attractiveness between infected and uninfected mice. Conclusion: In this first cross-sectional study with an aetiological agent of ACL, there were no statistically significant differences in the attractiveness of infected hosts to their vector. [ABSTRACT FROM AUTHOR]

The effectiveness of the MosqTent® trap was evaluated in endemic area to onchocerciasis in Brazil. This study seeks to provide subsidies for the monitoring of the onchocerciasis transmission in the country. The study was carried out at the Homoxi and Thirei villages, located in the Yanomami Indigenous Land, in the state of Roraima. This area presents hyperendemicity, high blackflies densities, large population migrations and mining activities. The Homoxi and Thirei villages are assisted by the Brazilian Ministry of Health. To conduct the present study, the village leader, health leaders and the Brazilian Ethics Committee were consulted. Blackflies captures were carried out simultaneously at the Homoxi and Thirei, using systematized methods to allow for comparisons between the traditional Human Landing Catch (HLC) and HLC protected by the MosqTent®. The female blackflies were captured at two equidistant capture stations per locality, by two collectors per station, for five consecutive days. Individuals captured by interval/station/day were counted, identified and maintained at -20°C. The underlying probability distributions and the differences between the methods for the independent sample data were verified in a comparative statistical analysis between the use of the MosqTent® and the HLC. A total of 10,855 antropophilic blackflies were captured by both methodologies. A total of 7,367 (67.87%) blackflies belonging to seven species were captured by MosqTent® —Simulium incrustatum s.l (99.06%); S. guianense s.l (0.74%), S. oyapockense s.l (0.01%), S. exiguum (0.10%), S. metallicum (0.05%), S. ochraceum (0.03%) and S. minusculum s.l (0.01%). Moreover, 3,488 (32.14%) blackflies belonging to four species were captured by HLC—S. incrustatum s.l (98.33%); S. guianense s.l (1.38%), S. oyapockense s.l (0.26%) and S. metallicum (0.03%). The MosqTent® was more effective and efficient when compared to HLC. When comparing total blackflies captured/day, the MosqTent® was more efficient than HLC (p = 0.031) with a means of 799.4 blackflies/day versus 217.6 blackflies/day by HLC. The results demonstrated improved performance and high reliability of the MosqTent® compared to the traditional HLC method. [ABSTRACT FROM AUTHOR]

More than 80% of available malaria rapid diagnostic tests (RDTs) are based on the detection of histidine-rich protein-2 (PfHRP2) for diagnosis of Plasmodium falciparum malaria. Recent studies have shown the genes that code for this protein and its paralog, histidine-rich protein-3 (PfHRP3), are absent in parasites from the Peruvian Amazon Basin. Lack of PfHRP2 protein through deletion of the pfhrp2 gene leads to false-negative RDT results for P. falciparum. We have evaluated the extent of pfhrp2 and pfhrp3 gene deletions in a convenience sample of 198 isolates from six sites in three states across the Brazilian Amazon Basin (Acre, Rondonia and Para) and 25 isolates from two sites in Bolivia collected at different times between 2010 and 2012. Pfhrp2 and pfhrp3 gene and their flanking genes on chromosomes 7 and 13, respectively, were amplified from 198 blood specimens collected in Brazil. In Brazil, the isolates collected in Acre state, located in the western part of the Brazilian Amazon, had the highest percentage of deletions for pfhrp2 25 (31.2%) of 79, while among those collected in Rondonia, the prevalence of pfhrp2 gene deletion was only 3.3% (2 out of 60 patients). In isolates from Para state, all parasites were pfhrp2-positive. In contrast, we detected high proportions of isolates from all 3 states that were pfhrp3-negative ranging from 18.3% (11 out of 60 samples) to 50.9% (30 out of 59 samples). In Bolivia, only one of 25 samples (4%) tested had deleted pfhrp2 gene, while 68% (17 out of 25 samples) were pfhrp3-negative. Among the isolates tested, P. falciparum pfhrp2 gene deletions were present mainly in those from Acre State in the Brazilian Amazon. These results indicate it is important to reconsider the use of PfHRP2-based RDTs in the western region of the Brazilian Amazon and to implement appropriate surveillance systems to monitor pfhrp2 gene deletions in this and other parts of the Amazon region. [ABSTRACT FROM AUTHOR]

Dengue is a major public health problem in tropical and subtropical areas worldwide. There is a lack of information on the risk factors for death due to severe dengue fever in developing countries, including Brazil where the state of Amazonas is located. This knowledge is important for decision making and the implementation of effective measures for patient care. This study aimed to identify factors associated with death among patients with severe dengue, in Amazonas from 2001 to 2013. We conducted a retrospective cohort study based on secondary data from the epidemiological surveillance of dengue provided by the Fundação de Vigilância em Saúde do Amazonas, FVS (Health Surveillance Foundation) of the Secretaria de Saúde do Amazonas, SUSAM (Health Secretariat of the State of Amazonas). Data on dengue cases were obtained from the SINAN (Notifiable Diseases Information System) and SIM (Mortality Information System) databases. We selected cases of severe dengue with laboratory confirmation, including dengue-related deaths of residents in the state of Amazonas from January 1, 2001, to December 31, 2013. The explanatory variables analyzed were sex, age, level of education, spontaneous hemorrhagic manifestations, plasma extravasation and platelet count. Patients who died due to severe dengue had more hematuria, gastrointestinal bleeding, and thrombocytopenia than the survivors. Considering the simultaneous effects of demographic and clinical characteristics with a multiple logistic regression model, it was observed that the factors associated with death were age >55 years (odds ratio [OR] 4.98), gastrointestinal bleeding (OR 10.26), hematuria (OR 5.07), and thrombocytopenia (OR 2.55). Gastrointestinal bleeding was the clinical sign most strongly associated with death, followed by hematuria and age >55 years. The study results showed that the best predictor of death from severe dengue is based on the characteristic of age >55 years, together with the clinical signs of gastrointestinal bleeding, hematuria, and low platelet count. [ABSTRACT FROM AUTHOR]

Background: Efforts have been made to establish sensitive diagnostic tools for malaria screening in blood banks in order to detect malaria asymptomatic carriers. Microscopy, the malaria reference test in Brazil, is time consuming and its sensitivity depends on microscopist experience. Although molecular tools are available, some aspects need to be considered for large-scale screening: accuracy and robustness for detecting low parasitemia, affordability for application to large number of samples and flexibility to perform on individual or pooled samples. Methodology: In this retrospective study, we evaluated four molecular assays for detection of malaria parasites in a set of 56 samples previously evaluated by expert microscopy. In addition, we evaluated the effect of pooling samples on the sensitivity and specificity of the molecular assays. A well-characterized cultured sample with 1 parasite/μL was included in all the tests evaluated. DNA was extracted with QIAamp DNA Blood Mini Kit and eluted in 50 μL to concentrate the DNA. Pools were assembled with 10 samples each. Molecular protocols targeting 18S rRNA, included one qPCR genus specific (Lima-genus), one duplex qPCR genus/Pf (PET-genus, PET-Pf) and one duplex qPCR specie-specific (Rougemont: Roug-Pf/Pv and Roug-Pm/Po). Additionally a nested PCR protocol specie-specific was used (Snou-Pf, Snou-Pv, Snou-Pm and Snou-Po). Results: The limit of detection was 3.5 p/μL and 0.35p/μl for the PET-genus and Lima-genus assays, respectively. Considering the positive (n = 13) and negative (n = 39) unpooled individual samples according to microscopy, the sensitivity of the two genus qPCR assays was 76.9% (Lima-genus) and 72.7% (PET-genus). The Lima-genus and PET-genus showed both sensitivity of 86.7% in the pooled samples. The genus protocols yielded similar results (Kappa value of 1.000) in both individual and pooled samples. Conclusions: Efforts should be made to improve performance of molecular tests to enable the detection of low-density parasitemia if these tests are to be utilized for blood transfusion screening. [ABSTRACT FROM AUTHOR]