Repositório RCAAP

In the last two years, a substantial increase in the number of malaria vivax cases has occurred in the Brazilian Amazon basin. The adequate exposure of hypnozoites to primaquine is a matter of interest as these dormant forms are responsible for the maintenance or even the increase of malaria burden in endemic areas. The aim of this study was to estimate the levels of primaquine and carboxyprimaquine in whole blood samples of patients with P. vivax treated with chloroquine and an abbreviated regimen of primaquine (0.5 mg/kg/d for 7 days), with adequate clinical and parasitological outcomes after 180 days of follow-up . A total of 40 male patients met the criteria for inclusion in the study. Primaquine and carboxyprimaquine were measured by high-performance liquid chromatography. The levels of primaquine in whole blood samples ranged from 40-238 ng/mL, 42-196 ng/mL and 42-150 ng/mL on days 1, 3 and 7. The levels of carboxyprimaquine in whole blood samples ranged from 87-234 ng/mL, 96-252 ng/mL and 74-448 ng/mL on days 1, 3 and 7. These data provide a reliable estimation of exposure of the infecting parasite to primaquine. Based on the regional pattern of relapse, the estimated blood levels of primaquine can be considered effective against hypnozoites of the local circulating strains of P. vivax.

Many snail species act as intermediate hosts of helminths that transmit diseases to humans and animals, such as schistosomiasis and angiostrongyliasis. São Gonçalo, a mostly urban municipality in Rio de Janeiro State, Brazil, has undergone fundamental environmental impacts, which favor the establishment of a range of diseases, for which snails act as the intermediate hosts of the etiological agents. In the present study, freshwater and terrestrial snail populations were surveyed in different environments within five city districts, and the presence of helminths was determined in the collected specimens. A total of 287 individuals were collected, six species from freshwater environment, Pomacea sp. (Ampullariidae), Melanoides tuberculata (Thiaridae), Biomphalaria tenagophila (Planorbidae), Dysopeas muibum (Subulinidae), Physa marmorata, and Physa acuta (Physidae), and two from terrestrial environment, Achatina fulica (Achatinidae) and Bradybaena similaris (Bradybaenidae). Snails were found in only two districts, Centro, an urban area, and Ipiiba, a rural area. Thirteen percent of the specimens of A. fulica eliminated larvae of the nematode Angiostrongylus cantonensis. None of the analyzed freshwater snails contained helminths.The most abundant and frequent snails were B. tenagophila, M. tuberculataand A. fulica, and the latter two species are exotic. The disturbance and degradation of natural areas adjacent to residential zones favor the proliferation of helminths, jeopardizing the local residents health. The abundance of A. fulica and B. tenagophila in the study area reinforces the need for a continuous and systematic monitoring of the snail fauna in this region.

Gut microbiota has been the subject of various molecular studies mainly due to its importance and wide-ranging relationships with human hosts. However, the storage of fecal samples prior to DNA extraction is critical when characterizing the composition of intestinal microbiota. Therefore, we aimed to understand the effects of different fecal storage methods to characterize intestinal microbiota using Next Generation Sequencing (NGS) as well as to establish an alternative conservation method of bacterial genetic material in these samples using guanidine. Stool samples from 10 healthy volunteers were collected. Each sample was divided into five aliquots: one aliquot was extracted immediately after collection (fresh) and two aliquots were subjected to freezing at -20 °C or -80 °C and extracted after 48 h. The other two aliquots were stored in guanidine at room temperature or 4 °C and extracted after 48 h. The V4 hypervariable regions of the bacterial and archeal 16S rRNA gene were amplified by PCR and sequenced using an Ion Torrent PGM platform for NGS. The data were analyzed using QIIME software. Statistical significance was determined using a nonparametric Kruskal-Wallis test. A total of 11,494,688 reads with acceptable quality were obtained. Unweighted principal coordinate analysis (PCoA) revealed that the samples were clustered based on the host rather than by the storage group. At the phylum and genus levels, we observed statistically significant differences between two genera, Proteobacteria (p=0.013) and Suterella (p=0.004), comparing frozen samples with guanidine-stored samples. Our data suggest that the use of guanidine can preserve bacterial genetic materials as well as freezing, providing additional conveniences

Background: The genetic variability of 610 S. aureus isolates from the hands of professional dentists (A), dental clinic environment air (B), bovine milk from cows with and without mastitis (C), an insufflator for milking equipment (D) and milking environment air (E) was studied by isoenzyme genotyping and genetic and cluster analysis.Results: Monoclonal and polyclonal patterns of S. aureus were detected in every bacterial population; however, isolates belonging to the same strain were not found among the populations, suggesting the genetic heterogeneity and the intrapopulation spread of strains. Genetic relationship analysis revealed the co-existence of highly related strains at low frequency among populations. Conclusion: The data suggest that some strains can adapt and colonize new epidemiologically unrelated habitats. Consequently, the occurrence of an epidemiological genotypic identity can assume a dynamic character (spread to new habitats), however infrequently. A tendency of microevolutionary and genetic divergences among populations of S. aureus from human sources (AB) and bovine milk (DE), and especially the mammary quarter (C), is also suggested. This research can contribute to the knowledge on the distribution and dissemination of strains and the implementation of control measures and eradication of S. aureus in important dental clinic environments, as well as animal environments and dairy production.

The objective of this study was to evaluate the presence of Lutzomyia longipalpis in the Argentine city of Concordia, in the province of Entre Rios, without record of this species despite previous samplings, but with canine cases of visceral leishmaniasis and Lu. longipalpis reports both, from Northern Argentine localities and from the city of Salto, Uruguay, located just across the river and the international border. This study confirms the presence of Lu. longipalpis and Nyssomyia whitmani, related to the risk of visceral and tegumentary leishmaniasis, respectively, in Concordia-Entre Rios. The presence of Lu. longipalpis confirms the ongoing dispersal along the Uruguay river basin. The presence of these species in the study area alerts about the risk of transmission of Leishmania spp.

Interferon-gamma (IFN-γ) release assays have improved latent tuberculosis (TB) detection and have been considered promising for the diagnosis of TB disease. However, diagnosis efficacy data is limited in high burden countries. The aim of this study was to determine the diagnostic potential of the QuantiFERON-TB Gold In-Tube (QFT-GIT) test for the diagnosis of active TB in an endemic setting for TB. A cross-sectional study was conducted in a group of 102 Thai patients with clinical symptoms and chest x-ray findings suggesting of active pulmonary TB and a group of 112 healthy adults. Testing was carried out using sputum microscopy, mycobacterial culture and QFT-GIT test. Of these patients, QFT-GIT was positive in 73 (71.57%), negative in 27 (26.47%), and undetermined in 2 (1.96%) cases. Among healthy controls, QFT-GIT was positive in 18 (16.07%), negative in 93 (83.04%), and undetermined in 1 (0.89%) person. Based on TB culture results, the sensitivity of QFTGIT for diagnosing active TB was 84.21% (95% confidence interval (CI); 72.13-92.52). The positive and negative predictive values were 65.75% (95% CI; 59.26-71.70) and 66.67% (95% CI; 49.94-80.04), respectively. The median IFN-γ level in culture-confirmed TB patients was 3.91 compared to 0.03 IU/mL of the healthy group. QFT-GIT appears to be a useful indirect test for TB diagnosis in Thailand and its use is recommended in association with clinical and radiological assessments for identifying active or latent TB.

The vasoactive intestinal peptide (VIP) expression is lower in cardiac chagasic patients and is related to worse cardiac function. The reduction of VIP in patients with Chagas disease may be a result of its enhanced degradation. To test this hypothesis, the tryptase and chymase expression was evaluated. We also related VIP levels with interleukin-17 (IL17) expression since VIP may modulate IL-17 production. Plasma levels of chymase were higher in chagasic patients. Conversely, VIP/chymase and VIP/tryptase ratios were lower in chagasic patients when compared to non-infected individuals. Besides, the VIP/chymase ratio was lower in chagasic cardiac patients in comparison with the indeterminate group. A positive correlation between tryptase and chymase levels was observed in chagasic cardiac patients. In relation to IL-17, we observed a higher expression of this cytokine in the cardiac form of the disease than in the indeterminate form. IL-17/VIP ratio was higher in the cardiac form in comparison with non-infected or indeterminate form. These results suggest that the low levels of VIP observed in chagasic patients could be due to an increased production of chymase and/or to the additive effect of the interaction between chymase and tryptase in the cardiac form. Moreover, the decreased VIP expression may contribute to the increase of IL-17 in chagasic cardiac patients.

Staphylococcus aureus (SA) is a commensal habitant of nasal cavities and skin. Colonization by community-acquired methicillin-resistant SA (CA-MRSA) is associated with infections in patients who have not been recently hospitalized. The aim of this study is to determine the prevalence of MRSA colonization in an outpatient population, currently unknown in Brazil. Three-hundred patients or caregivers from two teaching hospitals were included. A questionnaire was applied and nasal swabs were obtained from patients. Swabs were inoculated in brain heart infusion (BHI) with 2.5% NaCl and seeded in mannitol. Suspicious colonies were subjected to MALDI-TOF MS Microflex™ identification. Antimicrobial susceptibility test for oxacillin was performed for SA-positive samples by microdilution. Polymerase chain-reactions for detection of mecA and coA genes were performed for resistant samples. Data about MRSA carriers were compared with non-carriers. There were 127 S. aureus isolates, confirmed by MALDI-TOF. Only seven (2.3%) were MRSA and positive for mecA and coA genes. Factors associated with MRSA carriage were African ethnicity, skin diseases or antibiotic use. The majority of them were from Dermatology clinics. Prevalence of MRSA colonization in individuals from the community was low in our study (2.3%). This finding raises the hypothesis of inter-household transmission of SA, although we did not find any association between MRSA-colonization and the shared use of personal objects. Given the low prevalence of MRSA carriers observed, empirical antimicrobial coverage for MRSA in community-acquired infections should be not necessary.

Introduction: Candida parapsilosis is one of the main species that is able to adhere to forming biofilms on inert materials. Adhesion is the first step towards the colonization and invasion of host cells during the infectious process. Among the infections, vulvovaginal candidiasis is increasingly common. The objective was to evaluate the profile of adherence and biofilm formation of eight isolates of C. parapsilosis on the metal used in intrauterine devices (IUDs). Methods: Eight strains of C. parapsilosis presenting strong adhesion and biofilm formation properties were isolated from vaginal secretions in a previous study. To assay the adhesion and biofilm formation, copper fragments were made and cultivated in tubes containing 3 mL of phosphate-buffered saline and incubated for 6 and 24 h at 37 °C to evaluate biofilm formation. After incubation, the intensity of adherence and of biofilm formation on copper fragments were determined by performing a colony count. Results: All isolates were able to form biofilms and the isolate Cp62 showed many cells joined in a planktonic mode forming biofilms. The use of an IUD is one of the main factors that favors vulvovaginal candidiasis, and the presence of copper in this device increases the chance of recurrent vulvovaginal candidiasis (CVVR) due to the ease with which species of the genus Candida can adhere to inert surfaces. Conclusion: This research showed that the clinical isolates studied adhered to IUD copper fragments and formed biofilms, further increasing their virulence.

This study evaluated the antifungal activity and cytotoxicity profile of the ellagitannin punicalagin, a compound extracted from the L. pacari A.St.-Hil (Lythraceae) leaf, against Cryptococcus neoformans species complex. Minimum inhibitory concentrations (MIC) were checked using the broth microdilution method. Minimum fungicidal concentrations (MFC) and time of death were used to confirm the antifungal activity of the compound. The in vitro cytotoxicity of punicalagin was tested in BALB/c3T3 fibroblasts and A549 human lung cancer cell line, while the hemolytic potential was tested on sheep erythrocytes. The morphological changes induced in yeast strains by the presence of punicalagin were also analyzed. Tested on eight isolates of the C. neoformans complex punicalagin showed MIC of 0.5 to 4.0 μg/mL and MFC> 256 μg/mL. Punicalagin also demonstrated a good growth inhibitory activity in time-kill curves, but it was not able to achieve a statistically significant reduction of fungal growth suggesting a fungistatic effect of the compound. In vitro cytotoxicity studies using the two cell lines showed that punicalagin has low activity on these cells and no activity on sheep erythrocytes. Morphological changes were seen in the yeasts strains studied when treated with punicalagin. Therefore, punicalagin is a potential antifungal for important pathogenic yeasts and presents a low cytotoxicity profile associated with no hemolytic effects.

The Mitsuda test is a skin test based on the individual’s immune response through late and highly specific hypersensitivity reaction to the Mycobacterium leprae bacillus. A negative reaction identifies individuals who present a higher risk of becoming ill if exposed to M. leprae and, if they become ill, to develop the virchowian form of disease. The Mitsuda test reading is performed by means of a millimeter ruler. The dermatoscopy is a method that has not been used in the evaluation of cutaneous tests, although its use has increased in several areas. The study aimed to compare the results between the standardized reading and the total area of the Mitsuda test obtained by a computerized method which was structured by the association of digital dermatoscopy, the Dermatology Web system and the Image Tool 3.0 software. Data collection was performed at the Dermatology Outpatient Clinic of the Eduardo de Menezes Hospital, in Belo Horizonte, from November 2015 to August 2016. The sample consisted of 100 leprosy domiciliary contacts. There was an excellent agreement between the Mitsuda test (diameter and area), with a coefficient greater than 80%, and an excellent correlation with the Spearman’s correlation coefficient (0.936). The intraclass correlation coefficient indicated a low (0.219) but significant agreement between the two measurements. In conclusion, there is a significant correlation between the standardized reading and the total area of the Mitsuda test. Digital dermoscopy can be an alternative instrument of evaluation, allowing the computerization and recording of the Mitsuda test.

Chlamydia trachomatis (CT) infection is often silent and can lead to long-term reproductive complications in women. In this study, we determined the prevalence of CT infection and possible associations between the presence of the infection and clinicalepidemiological variables in pregnant women attended at the Basic Health Units of the Coari city, Amazonas, Brazil. From July 2016 to March 2017, 164 pregnant women undergoing prenatal care were recruited. One hundred of these women were tested for CT infection using two types of samples: cervico-vaginal and urine. The diagnosis was confirmed by PCR with primers specific for the omp1 gene of CT chromosomal DNA. Of the 100 pregnant women, 18 (18%) had CT infection, 8 (8%) of which were positive in both samples, 7 (7%) only in the urine sample and 3 (3%) only in cervical-vaginal sample. There was moderate agreement (Kappa=0.55) and no statistically significant difference between sample types (p = 0.400). The mean age of infected women was 21.1 years (SD = 4.6). Of the clinical-epidemiological variables analyzed, “more than 2 partners in the last 12 months” (p = 0.022) and gynecological complaint of “pain after intercourse” (p = 0.020) were associated with CT infection. This study showed a high prevalence (18%) of CT infection among pregnant women in Coari / Amazonas. Urine sampales were as good as cervical-vaginal ones for the screening of CT infection during the prenatal period.

Trypanosoma cruzi and Helicobacter pylori (HP) are pathogens that cause chronic diseases and have been associated with hypergastrinemia. The aim of this study was to evaluate the fasting gastrin levels in patients with different clinical forms of Chagas disease (CD), coinfected or not by HP. The enrolled individuals were outpatients attending at the university hospital. HP infection was assessed by serology and 13C-urea breath test. Fasting serum gastrin concentration was measured by chemiluminescence assay. Gastric endoscopic and histological features were also evaluated. Associations between CD and serum gastrin level were evaluated in a logistical model, adjusting for age, gender and HP status. A total of 113 patients were evaluated (45 with Chagas disease and 68 controls). In the multivariate analysis, increasing serum gastrin levels (OR= 1.02; 95% CI= 1.01-1.12), increasing age (OR= 1.05; 95% CI= 1.02 - 1.09) and HP-positive status (OR = 2.88; 95% CI = 1.10 - 7.51) remained independently associated with CD. The serum gastrin levels were significantly higher in the group of patients with the cardiodigestive form (P = 0.03) as well as with digestive form (P = <0.001) of Chagas disease than in the controls. In conclusion, patients with cardiodigestive and digestive clinical forms of CD have increased basal serum gastrin levels in comparison with controls. Moreover, we also demonstrated that H. pylori coinfection contributes to the hypergastrinemia shown in CD.

Mixed infection by Histoplasma capsulatum isolates with different mating types, in AIDS‑patients are described in this study. Morphological, mating type-specific PCR assay and multilocus sequencing type analysis of H. capsulatum isolates recovered from two Brazilian AIDS‑patients were performed. Five H. capsulatum isolates were recovered at different times from the two patients. Three isolates were obtained from bone marrow (day 1 – CE0411) and buffy coat cultures (day 1 – CE0311; day 2 – CE0511) of patient 1, and two isolates were isolated from buffy coat cultures (day 3 – CE2813; day 12 – CE2513) of patient 2. The mycelial colonies depicted different textures and pigmentation features. Dimorphic conversion to the yeast-phase in ML-Gema medium was achieved in all isolates. MAT1-1 idiomorph was identified in CE0311, CE0411 and CE2813 isolates; MAT1-2 idiomorph was found in CE0511 and CE2513 isolates. These H. capsulatum isolates were grouped within LAm A clade, highlighting that CE0311 and CE0411 isolates formed a subgroup supported by a high bootstrap value. The CE0511, CE2513, and CE2813 isolates clustered together with a Brazilian H151 isolate. This research reports mixed infections caused by H. capsulatum isolates with different mating types in Brazilian AIDS‑patients for the first time in the literature.

This study was designed to assess whether churches in endemic dengue districts in Merida, Mexico provide suitable breeding habitats for mosquitoes and are potential sites for dengue virus (DENV) transmission. Churches were inspected for immature and adult mosquitoes once every week from November 2015 to October 2016. A total of 10,997 immatures of five species were collected. The most abundant species were Aedes aegypti (6,051) and Culex quinquefasciatus (3,018). The most common source of immature Ae. aegypti were buckets followed by disposable containers. Adult collections yielded 21,226 mosquitoes of nine species. The most common species were Cx. quinquefasciatus (15,215) and Ae. aegypti (3,902). Aedes aegypti were found all year long. Female Ae. aegypti (1,380) were sorted into pools (166) and assayed for flavivirus RNA by RT-PCR and Sanger sequencing. Two pools were positive for DENV (DENV-1 and 2). In conclusion, we demonstrated that some churches in Merida are infested with mosquitoes all year long and they potentially serve as sites for DENV transmission and should therefore be considered for inclusion in mosquito and arboviruses control and surveillance efforts.

   

The diagnosis of Chagas disease is based on the detection of Trypanosoma cruzi (T. cruzi)-specific antibodies. Nonetheless, there is concern about the sensitivity of current serological assays due to reports of T. cruzi PCR positivity among seronegative individuals. The aim of this study was to evaluate if T. cruzi seronegative infections occur in endemic areas. We recruited 2,157 individuals that were identified as having Chagas disease in a public health system database of an endemic region in Brazil. All participants were interviewed and 2,091 had a sample collected for serological and PCR testing. From these, 149 (7.1%) had negative serological results. PCR was positive in 610 samples (31.4%) of the 1,942 seropositive samples but in none of the 149 samples from seronegative participants. True T. cruzi seronegative infections seem to be rare (95% CI 0-3.7) and should not be a concern for blood supply, which relies on antibody screening.

Chronic Hepatitis C relapse after liver transplantation can lead to graft failure within a short time period. The high efficacy and good safety profile of direct-acting antivirals has led to consensual recommendations for using interferon-free treatment after liver transplantation. However, pegylated interferon may still be required for genotype 3 non-responders. We treated a liver graft recipient with grade 1 fibrosis in the biopsy with daclatasvir and sofosbuvir for 12 weeks. He did not respond and progressed to grade 3 fibrosis. Lacking other options, we obtained a sustained virological response with pegylated interferon, ribavirin and sofosbuvir for 12 weeks. The combination of pegylated interferon, ribavirin and sofosbuvir is a viable option after the failure of direct acting antivirals in economically disadvantaged countries.

The present study sought to investigate the risk factors for physical disability upon release from multidrug therapy (MDT) in new cases of leprosy, registered at a referral center in Brazil. This is a longitudinal and retrospective study that evaluated 260 patients. Multivariate analyses, using both the ordinal logistic regression, as well as the classification and regression tree (CART) algorithm were performed to determine the factors associated with physical disability upon release from treatment. The prevalence of disability did not differ significantly between diagnosis and release from treatment. Number of affected nerves and sensory impairment upon diagnosis were risk factors for disability at the end of MDT. The analysis using the CART algorithm resulted in the development of a clinical score to predict the risk of disability upon release from MDT. The decision tree may have a direct applicability in clinical practice for professionals dealing with leprosy, as it allows them to identify patients with a higher risk of physical disability through the use of simple and widely available clinical tests. This study also shows that the disability grade upon admission is the main risk factor for disability upon release from MDT. This result draws attention to the importance of early diagnosis in disability prevention.

Consumption of ready-to-eat (RTE) vegetables is quick, easy and healthy, especially when eaten without cooking. However, they might be a source of foodborne pathogenic microorganisms. The objective of this study was to evaluate the microbiological and parasitological contamination of fresh RTE vegetables produced in agroindustries in the Federal District of Brazil (FD), and to correlate contamination with good manufacturing practices (GMP). One hundred and three samples of RTE vegetables were collected from six agroindustries for microbiology and parasitology analyses and correlate with GMP; 54 samples were collected from three hypermarkets for parasitological evaluation. None of the samples analyzed were positive for Salmonella sp. and for thermotolerant coliforms, but they were contaminated with total coliforms. All analyzed samples were contaminated with at least one species of enteroparasistes or commensals, which were identified as Ascaris sp., Balantidium coli, Entamoeba histolytica/dispar, Hookworm, Strongyloides sp., Trichuris sp., Entamoeba sp., eggs and larvae of Nematoda, insects and fungi. Agroindustries that adopted GMP showed less contamination with helminths. RTE vegetables sold in hypermarkets of the FD are unfit for human consumption. It is important to guide farmers in the FD on the need to adopt good practices in the production and processing of vegetables to reduce the microbial contamination.