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ABSTRACT In Fortaleza, the capital of Ceara State, Brazil, the detection rate of tuberculosis (TB) in 2018 was 65.5/100,000 inhabitants with a cure rate of 59.1%, which is higher than the country average. This study investigated the risk factors associated with drug-resistant tuberculosis (DR-TB) and identified the drug-resistance phenotype and resistance-conferring mutations. The geographic distribution of DR-TB in Fortaleza, Brazil, was also determined. From March 2017 to February 2018, 41 DR-TB isolates and 69 drug-susceptible pulmonary TB isolates were obtained from patients seen at a referral hospital in Fortaleza, Brazil. Samples were subjected to phenotypic and genetic analysis of resistance; the spatial distribution of the participants was also analyzed. Primary resistance was high (50.9%) among participants. The following risk factors for DR were identified: being female ( p = 0.03), having diabetes ( p < 0.01), history of previous TB disease ( p < 0.01), and the number of intra-domiciliary contacts ( p < 0.01). Analysis by multiplex allele-specific polymerase chain reaction detected mutations in the genes katG (65.8%) , rpoB (43.9%), inhA promoter (14.6%), and gyrA (9.8%). Sequencing identified mutations in the the genes katG (75.6%), inhA promoter (19.5%), rpoB (85.4%), and gyrA (100%). There was no mutation in the rrs gene. Spatial analysis showed DR-TB isolates distributed in areas of low socioeconomic status in the city of Fortaleza. Our results emphasized the importance of detecting resistance to TB drugs. The resistance found in the gene gyrA is of concern due to the high number of pre-extensive DR-TB cases in Fortaleza.

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ABSTRACT The aim of this study was to describe the microbiological characteristics and profile of genes encoding enterotoxins in 95 Staphylococcus aureus isolates obtained between April 2011 and December 2014 from foodstuffs, persons and surfaces of retail food stores. After microbiological identification and antimicrobial susceptibility testing, polymerase chain reactions (PCR) were performed, targeting sea, seb, sec, sed and see genes that code for classical enterotoxins (ET) A-E, and three additional genes: seg , seh and sei , coding for so-called “new enterotoxins” G, H and I. The isolates were characterized by Pulsed Field Gel Electrophoresis (PFGE), and five selected isolates were further analyzed through Multi Locus Sequence Typing (MLST). It is noteworthy that 54.7% of the examined isolates harbored one or more of the investigated ET gene types. Most positive isolates carried more than one ET gene up to five types; seg was the most frequent ET gene, followed by sei. Five enterotoxin-coding isolates also coded for some antimicrobial resistance genes. Two of them, and four additional non-enterotoxic isolates carried erm genes expressing inducible clindamycin resistance. PFGE-types were numerous and diverse, even among enterotoxin-coding strains, because most isolates did not belong to known foodborne outbreaks and the sampling period was long. MLST profiles were also varied, and a new ST 3840 was described within this species. ST 88 and ST 72 enterotoxin-coding isolates have been identified in other regions in association with foodborne outbreaks. This manuscript reports the first systematic investigation of enterotoxin genes in S. aureus isolates obtained from foodstuffs and infected people in Uruguay.

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ABSTRACT Most human epidemiological and clinical studies use visual inspection of the hair and scalp to diagnose Pediculus humanus capitis , however this method has low sensitivity to diagnose active infestations (presence of nymphs and adult lice). Vacuuming the hair and scalp has been used as a diagnostic method, but there are no previous data comparing its effectiveness with visual inspection. The aim of this study was to determine the prevalence of overall infestation (nits and trophic stages), of active infestation by Pediculus humanus capitis , and to evaluate the effectiveness of vacuuming in comparison with the visual inspection. Visual inspection was performed by three examiners and vacuuming of the scalp by one investigator, with an adapted vacuum cleaner. A total of 166 children aged 4 to 10 years old were randomly selected from public schools in Southern Brazil. Considering the positive results obtained by both methods, the prevalence of overall infestation was 63.3%, whereas active infestation was 18.7%. The visual inspection was more effective on diagnosing overall infestation, however, its effectiveness to detect active infestation was lower, ranging from 0.6% (RR=3%, p<0.001) to 6.6% (RR=35%, p=0.001), depending on the number of examiners. The effectiveness of vacuuming to diagnose active infestation was higher than the one of visual inspection, with a prevalence rate of 16.3% (RR=87%, p=0.332). As presented in our study, the vacuuming method was 2.74 to 7.87 times most likely to detect active infestation, thus it could be adopted as a more accurate method to diagnose active pediculosis.

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ABSTRACT Tuberculosis (TB) is still a leading cause of morbidity and mortality among people living with HIV (PLHIV). The diagnosis of latent TB is required for the implementation of prophylactic therapy with isoniazid (PTI). However, low access to diagnosis of latent TB and non-adherence to PTI may hinder potential benefits of this essential intervention. In this study, we addressed the access and adherence to PTI in a cohort of PLHIV with positive tuberculin skin test (TST) in a reference HIV clinic in Sao Paulo, Brazil. We have also analyzed the occurrence of active TB over a median of 131 months after a positive TST among study participants. Our findings revealed that 88.3% of the 238 TST-positive patients had access to PTI, and 196 (93.3%) of those with access adhered to PTI. Active tuberculosis was diagnosed in three of the 196 TST-positive patients who adhered to PTI (1.5%; 95% confidence interval [CI] 0.3-4.4%), whereas seven cases were detected among 42 patients without access or who did not adhere to PTI (16.6%; 95% CI 7.0-31.3%). The apparent beneficial effect of PTI in our cohort is consistent with previous studies including PLHIV, and highlights the importance of reliably delivering each of the steps between screening for latent TB and provision of PTI.

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ABSTRACT Aedes aegypti is associated with epidemic diseases in Brazil, such as urban yellow fever, dengue, and more recently, chikungunya and Zika viruses infections. More information about Ae. aegypti infestation is fundamental to virological surveillance in order to ensure the effectiveness of control measures in use. Thus, the present study aims to identify and compare infestation and infectivity of Ae. aegypti females in Macapa city, Amapa State (Amazon region), Brazil, between the epidemiological weeks 2017/02 and 2018/20. A total number of 303 Ae. aegypti females were collected at 21 fixed collection points, 171 at the 10 collection points in the Marabaixo neighborhood and 132 at the 11 collection points in the Central neighborhood. Among the collected samples, only two were positive for dengue virus, with a 2.08% (2/96 pools) infectivity rate for Marabaixo. The difference between the medians of Ae. aegypti females captured in Central and Marabaixo sites was not statistically significant. The findings indicate similar mosquito infestation levels between the neighborhoods, and a low-level of mosquito infectivity, although dengue virus was found only in Marabaixo. Virological surveillance of Ae. aegypti was important to identify sites of infection and determine possible routes of transmission to enable health surveillance teams to adopt preventive strategies where infected mosquitoes are present and act faster.

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ABSTRACT There are no studies in Mexico comparing Hepatitis C virus (HCV) epidemiology among Health Institutions. In this report, we described the deaths and hospitalizations due to HCV in the three main Health Institutions in Mexico: the Mexican Institute of Social Security, the Institute of Social Security for State Workers and the Ministry of Health, during the period 2004-2017. A secondary analysis was carried out across the country using hospital administrative death databases. Adult deaths and hospitalizations rates were calculated in reference to the total affiliated population and all-cause in-hospital mortality risk were also evaluated. There were 7,914 deaths and 9,002 hospitalizations due to HCV. Mortality and hospitalization rates of these three institutions together showed a continuous decrease over the analyzed time: the mortality rate dropped from 1.25 to 0.41 per 100,000 affiliates during 2004 and 2017, respectively (66.9% of change), and the hospitalization rate dropped from 2.19 to 0.39 per 100,000 affiliates (81.9% of change). All-cause in-hospital survival accounted for 89.6%. Older age groups and Ministry of Health hospitalizations were associated with higher all-cause in-hospital death rates. In conclusion, the mortality and hospitalizations rates found in this study reflect a decrease in the burden of HCV in Mexico.

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ABSTRACT Visceral leishmaniasis is a highly lethal zoonosis transmitted by a sandfly. It is caused by a Leishmania protozoan parasite and dogs are the main reservoir. Ceara State is endemic to visceral leishmaniasis and it is considered a high risk transmission area. Temporal and spatial epidemiological studies have been used as tools to analyze the distribution and frequency of human visceral leishmaniasis (HVL). This study aimed to characterize HVL in its epidemiological andtemporal aspects in Ceara State, from 2003 to 2017, as this is a neglected disease and a public health problem. This is an ecological study carried out with HVL confirmed cases in Ceara, using three blocks of years (2003 to 2007, 2008 to 2012 and 2013 to 2017). The disease presented an endemic behavior, affecting mainly male residents in the urban area, especially children under five and young adults between 30 and 49 years old. HVL is recorded in all the municipalities, for more than 10 years, with a growing trend and territorial expansion to the Central and Eastern regions of the State. The results of this study indicated the increase in the incidence and lethality, as well as the expansion of leishmaniasis in Ceara State.

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ABSTRACT For pregnant women with high viral load, antiviral therapy has been administered in addition to active and passive immune prophylaxis as a crucial adjunctive therapy to interrupt mother-to-child hepatitis B virus (HBV) transmission (MTCT). However, the time of antiviral therapy onset remains controversial. A systematic review and meta-analysis was conducted to compare the efficacy of antiviral therapy during the second or the third trimester for prevention of HBV vertical transmission. We searched nine databases for observational studies and randomized controlled trials that enrolled pregnant women with positive HBsAg treated with antivirals. The outcomes of interest were maternal HBV-DNA levels prior to delivery and the rates of HBV MTCT. We included nine studies that enrolled 1,502 pregnant women. The average HBV-DNA level before treatment was approximately 8 log10 copies/mL. Compared to the onset of antiviral intervention in the third trimester, the beginning of treatment in the second trimester distinctly reduced maternal predelivery HBV-DNA levels. However, no significant difference in HBV MTCT was found between the second and third trimester groups. Furthermore, the subgroup analysis showed that there were no significant differences between groups beginning treatment at different times (second or third trimester) with regard to HBV MTCT or other evaluated endpoints. For pregnant women with HBV-DNA levels less than or equal to 8 log10 copies/mL, the beginning of antiviral treatment can be delayed until the third trimester.

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ABSTRACT Proinflammatory and inflammatory mediators induced by Trypanosoma cruzi infection increase the oxidative stress, generating toxicity for cells targeting mitochondria of different tissues. We studied the activity of citrate synthase and complexes I-IV of respiratory chain in mitochondria of blood lymphomonocyte fraction, from albino Swiss mice infected with different isolates of T. cruzi , during Chagas disease evolution. Complexes I-IV were modified in infected groups (p<0.05) in all the stages, and an inflammatory process of different magnitudes was detected in the heart and skeletal muscle according to the isolate. The citrate synthase activity presented modifications in the SGO Z12 and the Tulahuen group (p<0.05). Hearts showed fiber fragmentation and fibrosis; skeletal muscle presented inflammatory infiltrates and in the Tulahuen infected group, there were also amastigote nests. The inflammatory processes produced an oxidative stress that induced different alterations of mitochondrial enzymes activities in the lymphomonocyte fraction that can be detected by a simple blood extraction, suggesting that they could be used as disease markers, especially in the indeterminate phase of Chagas disease.

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ABSTRACT Approximately 240 million people worldwide are infected by Schistosoma. In Brazil, one of the main intermediate hosts of this parasite is Biomphalaria glabrata snails. The early detection of larval stages in intermediate hosts is an important challenge to public health, but it also represents an opportunity as a new alternative to indicate earlier natural infections before cercariae differentiation and emergence. In this context, we demonstrated that PCR amplification of a 28S gene fragment from the parasite does demonstrate S. mansoni infection in snails 14 days post infection. This conventional polymerase chain reaction amplified clear bands and was able to detect parasitic infection in the intermediate host B. glabrata under experimental conditions. However, we reinforce that this approach requires deeper investigations and further comparisons to confirm its specificity and sensitivity in earlier time points after miracidia infection. This approach has relevant potential as an effective molecular-based strategy for the monitoring of schistosomiasis transmission.

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ABSTRACT Hepatitis B is a severe public health problem. The main world health centers have discussed it due to its pandemic proportion, high pathogenicity and infectivity. This study aimed to determine the immunization profile of dental surgeons - against hepatitis B virus (HBV) in the Public Health System (PHS) through the detection of anti-HBsAg antibodies by immunochromatography and associations with dental surgeons’ social and educational profile, factors related to professional knowledge and practices. This is a cross-sectional study on the prevalence of vaccination and immune status to HBV in dentists of Sao Paulo State PHS. Data collection occurred in three stages: questionnaire application, analysis of adhesion to the vaccination protocol and anti-HBsAg tests. Statistical analysis used the Bivariate Analysis and the Binary Logistic Regression. From the total of 219 interviewees, 74.9% reported having received three doses of the vaccine, but 35.6% were not immune to HBV. The dependent variable was associated with years in the public service (years) (OR [Odds Ratio]=1.04; 95% CI 1.00-1.08); knowledge on the subject (OR=6.93; 95% CI 1.39-34.40); incorrect answer concerning the etiological agent of the disease (OR=2.60; 95% CI 1.30-5.22); ignorance on the number of vaccine doses that have been administered (OR=3.43; 95% CI 1.14-10.30); and less than three doses of the vaccine in the immunization schedule (OR=8.76; 95% CI 3.50-21.91). A considerable portion of professionals non-immune to the HBV were found. We concluded that knowledge, years of practice and completion of the vaccination schedule (three doses) affected dental surgeons’ immune status to HBV.

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ABSTRACT The aim of study was to assess the prevalence of Treponema pallidum and HIV infection in Amerindian people (Mbya Guarani) over the age of 11 in Puerto Iguazu (Argentina) and to describe the contact tracking of cases. The method was a cross-sectional study in the Mbya Guarani people living in three villages of Puerto Iguazu (community A, pop. 1,146; community B, pop. 369; and community C, pop. 149). Participants were randomly invited to participate in the survey and in blood testing. Of the 551 participants, 48 were infected by T. pallidum (8.71%; 95% confidence interval [CI] 7.38-10.04). The infection prevalence decreased with age, standing at 9.66% in the 11-19 age group, 8.42% in 20-39 age group and 4.54% in people aged 40 and older. We tracked 130 contacts for the 48 T. pallidum cases; 39/40 (97.5%) sexual contacts tested positive for syphilis. Among the 90 children born to infected mothers, 76 aged 18 months or older tested negative, while 8/14 younger children were still at risk for congenital syphilis. There were four cases of HIV infection (0.72%, 95% CI 0.31-1.13). Prevalence of T. pallidum infection and HIV infection are relevant in this indigenous community of Argentina, representing a public health concern.

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ABSTRACT Giardia duodenalis is one of the most important and widespread gastrointestinal parasites in the world. Despite its relevance as a causative agent of diarrhea, asymptomatic giardiasis occurs frequently, especially in low resources settings in which children are exposed to many risk factors. Based on microscopic examination and the polymerase chain reaction (PCR) amplification and sequencing of beta-giardin (bg), triose phosphate isomerase (tpi) and glutamate dehydrogenase (gdh) genes, we assessed G. duodenalis occurrence and genetic diversity in isolates of children attending a daycare center and living in low income families, in an economically successful region. Considering both, microscopic examination and PCR/sequencing methods, the overall prevalence of Giardia infection was 51.4%, with the highest frequency in children aged 1-4 years old (p<0.05). Genotyping of 50 isolates revealed that the assemblage A was found in 60% of the samples (30/50), followed by the assemblage B in 38% (19/50) and 2% of mixed-assemblage infections (1/50). At the sub-assemblage level, isolates genotyped as A were AII and among isolates B, BIII and BIV were identified. Both assemblages A and B were detected in children of all age groups, however assemblage A was more prevalent. The detection of anthroponotic assemblages and sub-assemblages (AII, BIII and BIV) reinforces human-to-human transmission, mainly in children of all age groups when they have not yet received toilet training, making them more vulnerable to infection.

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ABSTRACT The aim of this study was to determine the spontaneous decolonization period and characteristics in a prospective cohort of newborns colonized by multidrug-resistant organisms, after their discharge from the neonatal intensive care unit. Multidrug resistance is defined as bacterial non-susceptibility to ≥ 1 agent of ≥ 3 antimicrobial categories. In total, 618 newborns were included in the study, of which 173 (28.0%) presented a positive culture for multidrug-resistant microorganisms, and of these, 52 (30.1%) were followed up in this study. The most frequent intrinsic factors were be born by cesarean section (86.5%), prematurity (84.6%), and very low birth weight (76.9%). The extrinsic factors were having remained hospitalized for an average of 27 days, during which 67.3% were submitted to invasive procedures and 88.5% received antimicrobials. The intrinsic and extrinsic factors of newborns were not associated to a decolonization period longer or shorter than 3 months, which was the average period of decolonization found in the present study. From the totality of colonization cultures sampled at hospital discharge, the Gram-negative Extended Spectrum β-lactamase producing bacteria were the most common, with 28.9% of babies colonized by Klebsiella spp. The median period of decolonization by multidrug-resistant microorganisms in the newborns population after hospital discharge was 3 months, but was highly dependent on the microbial species, and this period was not associated to any intrinsic and extrinsic factors of the newborn.

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ABSTRACT Candida is one of the most frequent pathogens of bloodstream infections, which is associated with high morbidity and mortality rates. Rapid immunological detection methods are essential in the early diagnosis of candidemia. Anti-mannan is one of host-derived biomarkers against cell wall components of Candida. We conducted this study to evaluate the diagnostic performance of two anti-mannan assays (IgM, IgG) for candidemia through the analysis of 40 candidemia patients, 48 participants with Candida colonization and 213 participants with neither Candida colonization nor Candida infections (13 patients with other bloodstream infections, 145 hospitalized patients and 55 healthy controls). The performance of the two assays were evaluated by calculating their sensitivity and specificity. The sensitivity ranged from 0.78 to 0.80 for the IgM assay and 0.68 to 0.75 for the IgG assay. The specificity ranged from 0.97 to 0.98 for the IgM assay and 0.91 to 0.94 for the IgG assay. The diagnostic performance of the anti-mannan IgM assay was better than that of IgG, with higher sensitivity and specificity. Combining the two assays (positive results of single or both assays are both considered as positive) could improve the sensitivity up to 0.93 (0.79-0.98) and only slightly reduce the specificity (0.93(0.89-0.95)). The anti-mannan IgM, IgG assays are rapid and cost-effective assays that may be probably useful in the diagnosis of candidemia.

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ABSTRACT Scorpionism has a high incidence rate in Brazil. It is considered a serious public health problem mainly in tropical and subtropical regions around the world. The number of scorpion accidents have increased over the years and the highest frequencies have been reported mainly in the Brazilian Northeast region. Therefore, in this study we report a retrospective clinical and epidemiological analysis of scorpion stings from 2007 to 2017 in Alagoas State, Northeast Brazil, at a referral hospital for assistance and treatment of accidents by venomous animals. During the analyzed period, the referral hospital treated 27,988 cases, and an increase in the number of cases has taken place over the years. The highest frequency of scorpion stings was observed in females, and the age range most affected was from 20 to 29 years old. The most stung body site was the foot, followed by finger, toe or hand. Regarding the severity, most severe cases were reported in children up to 4 years old (69.4%) and 50% of the total cases treated with serotherapy corresponded to patients in this age range. Interestingly, it was also found that the occurrence of systemic manifestations and the severity of the cases were significantly associated with pediatric patients. In this way, this study highlights the scorpionism as an environmental public health problem in Alagoas State, Northeast Brazil, as well as the need to intensify the epidemiological surveillance and educational campaigns to prevent and control scorpion accidents throughout the year.

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ABSTRACT Identifying the true prevalence of human T-cell lymphotropic virus, mostly type 1 (HTLV-1), and the number of patients with HTLV-1-associated diseases, in addition to introducing HTLV-1/2 serology during the prenatal of pregnant women and in individuals infected with other viruses that share transmission routes with HTLV-1, are actions that could help to recognize the importance of this virus by WHO and national health organizations, and to control its transmission/dissemination. As Brazil is endemic to HTLV and there is an increase in health care expenditure, but resources are limited, any strategy that could reduce the cost of HTLV screening is needed and welcomed. This study aimed to determine whether the strategy of pooling sera for HTLV antibody determination is feasible and reduces the costs. Two enzyme immunoassays (EIA Murex HTLV-I+II, Diasorin, UK, and Gold ELISA HTLV-1+2, REM Ind. Com. Ltda., SP, Brazil), and serum samples that resulted in different levels of HTLV-1/2 antibodies by EIA and of which a volume allowed assay validation were employed for analysis. The diagnostic sensitivity and specificity and Cohen’s Kappa value, as well as the accuracy and precision were analyzed. After validating the five-sample pool using the EIA Murex (Cohen’s Kappa = 1.0), the technique was employed for individual cost comparison in 2,625 serum samples from populations at risk of HTLV infections (HBV, HCV, and HIV-infected individuals). The results from individual and pooled samples confirmed the diagnostic sensitivity (100%) and specificity (100%) of the pooling and a cost minimization varying from 60.7% to 73.6%. In conclusion, the results of this study suggest the use of pooling sera in sero-epidemiological surveillance studies and possibly in prenatal care screening programs in Brazil.

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ABSTRACT Immunosuppression is an important risk factor for leishmaniasis. We assessed the clinical profile, geographic distribution and prevalence of leishmaniasis in patients undergoing immunosuppressive therapy for dermatological, rheumatological or gastroenterological autoimmune diseases. We identified relevant studies in PubMed, EMBASE, Scopus, Web of Science and LILACS on July 3rd, 2018. We included articles that reported at least one case of leishmaniasis in patients undergoing immunosuppressive treatment for dermatological, rheumatological or gastroenterological diseases. Our protocol was registered in PROSPERO (CRD42018103050). We assessed the quality of the included studies with the Joanna Briggs Institute Critical Appraisal Tool. After the removal of duplicates, 5,431 articles were collected and screened. We included 138 articles; the prevalence of leishmaniasis in six methodologically similar studies varied from three to 1,282 cases per 100,000 patients using anti-TNFα drugs, but the results were significantly heterogeneous . Leishmaniasis in patients treated with immunosuppressive drugs is a health problem mostly reported in European countries bordering the Mediterranean Sea; sporadic activities, such as travelling, seem not to be associated with a significant risk of leishmaniasis, although effective control measures must always be observed.

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ABSTRACT We conducted the genome sequencing and analysis of the first confirmed COVID-19 infections in Brazil. Rapid sequencing coupled with phylogenetic analyses in the context of travel history corroborate multiple independent importations from Italy and local spread during the initial stage of COVID-19 transmission in Brazil.

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ABSTRACT Oral problems are common in patients diagnosed with Eating Disorders (ED) and still require better elucidation. We aimed to analyze the prevalence of oral Candida spp in individuals with ED. The sample of the study was comprised of 30 women with purgative habits and 15 without purgative habits. Samples of the oral cavity were collected by sterile cotton swab rubbed on soft tissues and teeth. Yeasts were isolated on Sabouraud dextrose agar. Yeasts were isolated from the oral cavity of 53% of the patients yielding 75 yeast isolates; of these, 43 were identified by conventional mycological methods: C. parapsilosis (n=19), C. glabrata (n=16), Rhodotorula sp (n= 6), C. famata (n=2). The remaining 32 isolates were presumptively identified as C. albicans or C. dubliniensis and required mass spectrometry for the final differentiation: 28 isolates were confirmed as C. albicans and four as C. dubliniensis. Among the control group, only four subjects (26.7%) were found to harbor C. albicans. The four C. dubliniensis isolates were from two patients, one that was only colonized and the other, with severe ED, was diagnosed with an oral candidiasis as demonstrated by the presence of pseudohyphae on the direct mycological exam from different sites. The increased rate of isolation of non-albicans species, such as C. glabrata, C. parapsilosis, and C. dubliniensis in the oral cavity from ED patients with nutritional deficiency may suggest that purgative habits of these patients can lead to changes in normal flora and predispose to oral candidiasis.

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ABSTRACT Simple, low-cost and effective diagnostic tests for tuberculosis (TB) are needed especially in TB-high burden settings. The present study evaluated the performance of an in-house loop-mediated isothermal amplification (LAMP) for diagnosing TB by comparing it to Xpert MTB/RIF, microscopy and culture. In Thailand, a total of 204 excess sputum samples volume after the processing of cultures were used for Mycobacterium tuberculosis (MTB) detection by Xpert MTB/RIF and LAMP. Based on culture results as the gold standard, the overall sensitivity of LAMP and Xpert MTB/RIF were 82.1% (126/153; 95% confidential interval [CI]: 75.4-88.98%) and 86.9 % (133/153; 95% CI: 80.5-90.8%) respectively, and the specificity of both tests was 100% (51/51; 95% CI: 93.0-100.0%). In comparison with Xpert MTB/RIF, the sensitivity and specificity of LAMP were 94.7% (126/133; 95% CI: 89.5-97.9%), and 100.0% (73/73; 95% CI: 94.9-100.0%), respectively. The average threshold cycle (Ct) of Xpert MTB/RIF detection for positive and negative LAMP results was statistically different, of 18.4 and 27.0, respectively (p < 0.05). In comparison with the acid-fast staining technique, and analyzing LAMP and Xpert MTB/RIF in smear-negative/culture-positive specimens, there was an increase of the detection rate by 47.7% (21/44) and 54.6% (24/44). The diagnostic sensitivity and specificity of LAMP appeared to be comparable to those of Xpert MTB/RIF. We claim that this LAMP has potential to provide a sensitive diagnostic test for the rapid TB diagnosis. It allowed a fast detection of MTB before the cultures and it could be used in resource-limited laboratory settings.

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ABSTRACT Drug resistant tuberculosis (DR-TB) is challenging particularly in developing countries. As such, a previous investigation gave the first insight into the mutational status of the Rifampicin Resistance Determining Region (RRDR) of rpoB gene among a restricted number of MTB patients’ residents in the Northern Morocco. The purpose of this study was to investigate rpoB mutation types and frequencies associated with resistance to Rifampicin in a larger panel of MTB patients and to evaluate the usefulness of these mutations to improve the diagnosis of resistance to Rifampicin. A panel of 301 consecutive sputum samples belonging to patients suscpected of having TB from Northern Morocco was collected at the Pasteur Institute of Tangier between 2014-2017. Samples were subjected to conventionel microbiological tests. Evaluation of rpoB muational status was assessed by PCR amplification and sequencing of the RRDR of the rpoB gene. DST results showed that 26.4% of strains were MDR. Sequencing results reported single point mutations in 36 of 65 RIFR isolates of which two had two mutations. Aminoacid substitutions in the codon Ser531Leu occurred at the highest frequency (34.46%). Overall, 10 aminoacid substitutions have been registered, and the H526S substitution was reported for the first time. The present study highlighted that resistance to RIF is a reliable marker of MDR-TB, the common mutations successfully detected in the rpoB 531, rpoB526 and rpoB516 codons provide a foundation for the implementation of molecular approaches such as Hain and GeneXpert as a routine tests to detect DR-TB. However, considerable work is still necessary to identify extensive mutations associated with DR-TB.

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ABSTRACT In this study which is part of a research project on Chagas disease (CD) among Bolivian immigrants in Sao Paulo, we describe socioeconomic characteristics, knowledge of CD and implications for acess to health care. We applied a structured questionnaire to a sample of 472 Bolivian adults (> 18 years) living in Sao Paulo and enrolled at the Barra Funda School Health Center. Participants’ median age was 28.5 years, 75.0% were from the Bolivian department of La Paz, and >90% worked in the garment industry. Respondents had lived in Sao Paulo for a median of 5.8 years. Only 169 (35.8%) were familiar with CD, while roughly half (50.4%) had lived in natural materials houses in Bolivia, 225 (47.7%) indicated familiarity with the vector, 23.9% had seen the vector in their homes in Bolivia, and 6.4% reported having been bitten by a triatomine bug. Factors associated with awareness of CD were analyzed by chi square tests, and those with p values <0.25 were included in a multivariable logistic regression model. In the multivariable logistic regression analysis, having a relative with CD (OR=4.3, 95% CI=1.5-12.0), having lived in a house with mud or wood walls (OR=0.4, 95% CI=0.2-0.8), and having heard of the triatomine bug, or vinchuca, (OR=10.0, 95% CI=5.1-19.5) were significantly associated with awareness of CD. This study shows a low familiarity with CD among Bolivian migrants living in Sao Paulo, Brazil. Raising awareness of the disease through specific communication strategies should be an essential component of public health programs to reduce the burden of CD in this and other vulnerable populations.

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ABSTRACT The new coronavirus, COVID-19 was declared a pandemic by the World Health Organization on March 11, 2020. Risk factors associated with this disease are age, sex, and the presence of comorbidities, the most common being hypertension, diabetes, and heart disease. The aim of this meta-analysis was to calculate the prevalence and geographical distribution of comorbidities in all patients admitted to intensive care units (ICUs), and the mortality rate of COVID-19. We selected studies based upon epidemiological and clinical descriptions of the patients and mortality from the disease to determine the pooled prevalence of comorbidities in all patients and in mortality cases due to COVID-19. The pooled prevalence was estimated using the random effects model, and odds ratios were used to measure the probability of death for a patient with a comorbidity. The total prevalence of comorbidities in patients with COVID-19 was 42% (95% CI: 25-60), 61% (95% CI: 42-80) in those admitted to the ICU, and 77% (95% CI: 68-86) among death cases; males were the most affected. Hypertension was the most prevalent comorbidity in all three groups studied, accounting for 32%, 26%, and 35%, respectively. The odds ratio of death for a patient with a comorbidity compared to one with no comorbidity was 2.4 (P < 0.0001). The higher the prevalence of comorbidities the higher the odds that the COVID-19 patient will need intensive care or will die, especially if the pre-existing disease is hypertension, heart disease, or diabetes.

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ABSTRACT Infection by the protozoan Toxoplasma gondii during pregnancy demands greater attention from the health authorities due to the risk of placental transmission, which can have devastating consequences to the foetus and newborn. This study was conducted in a high-risk prenatal care outpatient clinic of a university teaching hospital. Pregnant women screened for specific IgM and IgG anti -T. gondii, attended from January 2009 to August 2018 were included. From 530 suspected patients, 218 were followed up and they presented positive IgM and IgG anti- T. gondii. From these patients, 83 (38.0%) had low IgG avidity, 39 (18%) seroconverted in the second or third trimester of pregnancy, 19 (8.7%) had no avidity test, 69 (31.6%) had high IgG avidity after 16 weeks of gestation, five had recurrent chorioretinitis (2.2%) and three (1.3%) were seropositive to HIV. Complementary diagnoses were made in 30/48 (62.5%) of the patients revealing the presence of specific IgA antibodies raised to T. gondii; 3/63 (4.8%) peripheral blood samples and 1/57 (1.8%) amniotic fluid sample. There were eight foetal deaths, one case of neonatal hepatomegaly and one case of T. gondii DNA detected in a peripheral blood sample. Of the 139 newborn deliveries at the teaching hospital, there was a 38% loss of follow-up. The prevalence of congenital toxoplasmosis was 1.2 cases/1,000 live births in this study area, according to the retrospective survey of cases. Prenatal treatment may have helped to reduce the risk of vertical transmission.

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ABSTRACT Efficient snail production is essential for the proper maintenance of the Schistosoma mansoni life cycle in the laboratory. In order to improve the breeding of Biomphalaria glabrata under laboratory conditions, this study aimed to demonstrate the effectiveness of a handmade shell-based substrate on the physiological performance of B. glabrata. The shells used to make the substrate were cleaned, sterilized and macerated until a powder was obtained (yield of 92.3%). B. glabrata specimens were randomly assigned to three treatment groups: negative control group (NCG) exposed to a clay-only substrate; a positive control group (PCG) containing clay, oyster flour and calcium carbonate; and the test group (TG) with the shell-based substrate and clay. B. glabrata bred in the test group showed improved growth, sexual maturity, fertility, mortality rate, and shell morphology when compared to the NCG, and similar to the PCG. Therefore, the shell-based substrate proved to be efficient and has a low cost for the breeding of B. glabrata.

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ABSTRACT This study analyzed possible associations between the trimester of maternal Zika virus infection (ZIKV) in pregnancy, severity of brain computed tomography (CT) findings and the presence of microcephaly at birth in children with Congenital Zika Syndrome (CZS). It was an analytical study in a cohort of children with CZS. Symptoms of maternal infection were dichotomized into the 1st trimester of pregnancy and other trimesters. Head circumference (HC) at birth was used to calculate the z-score. Mild microcephaly was defined as HC between 2 and ≥3 standard deviations (SD) below the mean for each gestational age and sex, and severe microcephaly when HC <3 SD below average. Brain CT images were evaluated by two radiologists and classified, according to the severity, into mild, moderate and severe. Fisher’s exact, Mann-Whitney and Kruskal-Wallis tests were used to verify the associations between variables. In 108 children, maternal infection in the 1st trimester of pregnancy was associated with more severe brain CT abnormalities (p=0.038), greater severity of microcephaly at birth (p=0.013) and lower HC z-scores at birth (p=0.021). The severity of brain CT lesions was also associated with lower HC z-scores at birth (p<0.001). Maternal ZIKV infection during the first trimester of pregnancy proved to be an important risk factor for a more severe spectrum of CZS, as it is associated with more severe brain CT abnormalities and, consequently, with lower HC z-scores at birth.

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ABSTRACT This study aimed to describe the spatial distribution and assess entomological indicators of synanthropic triatomines in Piaui State, Northeastern Brazil. We used surveillance data on the detection, identification and assessment of natural infection with trypanosomatids from triatomines in the State from 2014 to 2017. The State was divided into four macroregions. In relation to the dispersion rates of triatomines, they were much lower in the North, when compared to Southwest, Southeast and Central North macroregions. Infestation rates were higher in the Southwest and Southeast and intradomicile infestation rates varied during the study period, reaching high values in all regions. Insects belonging to the species Triatoma brasiliensis complex, Triatoma pseudomaculata, Triatoma sordida, and to the genus Panstrongylus spp. and Rhodnius spp. were collected during this period. T. brasiliensis was collected from all four regions of the State, but more frequently in those located in the Southeast. A similar pattern was observed for T. pseudomaculata. T. sordida was detected in the municipalities in the Southeast and Southwest regions, and less frequently in the Central North municipalities. Rhodnius spp. was detected in the Central North and North regions, and Panstrongylus spp. in the Central North and Southeast regions. The highest trypanosomatid-positivity rate of T. brasiliensis and Panstrongylus spp. was in the Southeast region. A significant proportion of the municipalities of Piaui State presents entomological parameters that indicate a risk of Chagas disease by vector transmission.

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ABSTRACT An outbreak of bloodstream infection (BSI) caused by members of the Burkholderia cepacia complex (Bcc) took place from March 2012 until April 2014 involving thirteen patients. Aim. To describe an outbreak investigation of BSI Bcc and showing how genetic sequencing tools contributed to confirm the hypothesis of extrinsic contamination proposed by an observational study. Methods. The Infection Control Department revised and reinforced good practices of infusion therapy and catheter care, visits to affected wards, a case control study, and environmental screening based on the case-control findings. Results. Data from the case-control study found an association of cases with central venous catheter (OR 1.36; CI 1.15-1.67) and intravenous cisatracurium use (OR 10.75; CI 1.67-68.89). Visits to the operatory block revealed problems related to the cold chain used for the preservation of thermolabile cisatracurium. We could not retrieve Bcc from environmental samples using classic microbiology. New samples from the same surfaces were obtained for genetic sequencing. Bcc was identified in the cooler box, refrigerator and reusable ice packages. Conclusion. Environmental screening using genetic sequencing proved to be a useful tool for confirming our hypothesis of extrinsic contamination raised by the case-control study.

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ABSTRACT Intestinal mucins are the first line of defense against microorganisms. Although knowledge about the mechanisms involved in the establishment of intestinal protozoa is limited, there is evidence that these parasites produce lectin-like molecules and glycosidases, that exert both, constitutive and secretory functions, promoting the establishment of these microorganisms. In the present review, we analyse the main interactions between mucins of the host intestine and the four main protozoan parasites in humans and their implications in intestinal colonization. There are lectin-like molecules that contain complex oligosaccharide structures and N-acetylglucosamine (GlcNAc), mannose and sialic acid as main components, which are excreted/secreted by Giardia intestinalis, and recognized by the host using mannose-binding lectins (MBL). Entamoeba histolytica and Cryptosporidium spp. express the lectin galactose/N-acetyl-D-galactosamine, which facilitates their adhesion to cells. In Cryptosporidium, the glycoproteins gp30, gp40/15 and gp900 and the glycoprotein lectin CpClec are involved in protozoan adhesion to intestinal cells, forming an adhesion-attack complex. G. intestinalis and E. histolytica can also produce glycosidases such as β-N-acetyl-D-glucosaminidase, α-d-glucosidase, β-d-galactosidase, β-l-fucosidase, α-N-acetyl-d-galactosaminidase and β-mannosidase. In Blastocystis, α-D-mannose, α-D-glucose, GlcNAc, α-D-fucose, chitin and sialic acid that have been identified on their surface. Fucosidases, hexosaminidases and polygalacturonases, which may be involved in the mucin degradation process, have also been described in the Blastocystis secretoma. Similarly, symbiotic coexistence with the intestinal microbiota promotes the survival of parasites facilitating cell invasion and nutrients obtention. Furthermore, it is necessary to identify and characterize more glycosidases, which have been only partially described by in silico analyses of the parasite genome.

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ABSTRACT The aim of the study was to assess the factors associated with mother-to-child transmission (MTCT) of HIV. The study design is a retrospective cohort. The population consisted of 323 HIV-positive mothers and their newborns, attended at the Perinatal Nucleus/HUPE-UERJ, municipality of Rio de Janeiro, in the period of 2007-2018. The average age of mothers was 27 years (14-44), with 12.7% (41) of adolescents. The majority (66.8%) knew they were infected during pregnancy: 39.4% in the current pregnancy and 27.4% in a previous pregnancy. The incidence of MTCT was 2.7% in 2007-2009, 1% in 2010-2015 and 0 in 2016-2018. The viral load in the 3rd trimester of pregnancy was > 1.000 copies/mL or unknown in all mothers with positive newborns and in 19% (42/221) of mothers with negative newborns (p=0.003). The duration of antiretroviral use was > 4 weeks in 92.3% (264/286) of mothers with HIV-negative newborns and in 2 in the HIV-positive group (p=0.004). One of the 4 infected newborns and 2 of the negative ones did not use oral zidovudine (p=0.04). There was no association between amniorrhexis and MTCT (p=0.99), with the Apgar score in the 5th minute of life (p=0.96), with marital status (p=0.54), ethnicity (p=0.65), adolescence (p=0.42), mode of delivery (p=0.99), beginning of prenatal care (p=0.44) or with maternal comorbidities (p=0.48). The conclusion of the study points out that the main factors associated with MTCT are the elevated maternal viral load in the 3rd trimester, the time of use of ART and the non-administration of zidovudine for the newborns.

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ABSTRACT Asymptomatic VL is a concern, considering the risk of transmission in highly endemic areas due to human-to-human transmission. The aim of this study was to report the sero-epidemiological prevalence in Bihar, India, a highly endemic area of VL, using the leishmanin skin test (LST) and the direct agglutination test (DAT). This was a cross-sectional study performed in Muzaffarpur, Bihar, India. Relatives of patients with VL were tested by LST and DAT. Other epidemiological data were evaluated and correlated with tests results. Forty individuals (either previous or current patients), and 109 household contacts were studied. There were 36% of male visceral leishmaniasis family members versus 17.57% of females visceral leishmaniasis family members, thus showing more males with symptomatic disease than females (p< 0.01). All visceral leishmaniasis cases had positive DAT tests, but only 37% of past cases were positive on the skin testing. Amongst healthy household contacts, 34% were DAT-positive, whilst 21% were LST-positive. The overall positivity for both assays combined was 44.8% and 23.8% were DAT-positive alone. The finding of high infection prevalence amongst asymptomatic individuals, and the estimation of those at greater risk for overt disease (DAT-positive alone) are important in the development of future disease control policies.

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ABSTRACT The Northeast of Brazil has experienced a triple epidemic, with the simultaneous circulation of dengue virus (DENV), chikungunya virus (CHIKV) and Zika virus (ZIKV), which may have contributed to the observed increase across this region of atypical forms of disease and deaths. In view of this fact, non-congenital neurological disorders related to arboviruses were compared with other etiologies, mortality and survival rates of patients admitted to referral neurology hospitals in Pernambuco State, Northeast Brazil, from 2015 to 2018. Blood and cerebrospinal fluid samples were collected and tested using molecular and serological assays. The arbovirus-exposed groups were compared with respect to epidemiological, clinical and neurologic characteristics by using the Pearson’s chi-square test. For the survival analysis, the Kaplan-Meier and Hazard Ratio (HR) tests were used, with a 95% confidence interval (CI). Encephalitis and encephalomyelitis were more frequent in arboviruses, while myelitis predominated in the neurological disorders of other etiologies. Guillain-Barré Syndrome (GBS) was similarly distributed amongst the groups. Exposure to one of the arboviruses caused a six-fold increase in the risk of death (HR: 6.37; CI: 2.91 - 13.9). Amongst the arbovirus-exposed groups, infection (DENV/CHIKV) increased nine times the risk of death (HR: 9.07; CI: 3.67 - 22.4). The survival curve indicates that have been exposed to some arbovirus decreased the likelihood of survival compared to those with other etiologies (Log-Rank: p<0.001). Within this scenario, neurologic manifestations of DENV, CHIKV and ZIKV have the potential to increase mortality and decrease survival, and concomitant infection (DENV/CHIKV) is an aggravating factor in reducing the likelihood of survival when compared to monoinfections.

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ABSTRACT The study aimed to analyze the impact of health interventions carried out in the city of Palmas, Brazil, on the epidemiological and operational indicators of leprosy between 2007 and 2017. The intervention consisted of training healthcare personnel on the diagnosis and follow-up of patients and organizing the referral of patients to health units. Estimates of the impact were calculated by taking the differences between indicators reported in two equal periods of 1.5 years pre- and post-intervention, with a transition period of six months. During the study period, the database contained 1,875 notifications, with 66% of cases diagnosed in the post-intervention period. There was a predominance of males (52%); aged 50 years or more (34.9%); with mixed ethnicity (63.5%). The low level of education was noticeable, with more than half of the cases (51.7%) reporting illiteracy or ≤ 7 years of education. The intervention resulted in an increase in both, epidemiological and operational indicators, suggesting a positive impact of the intervention on leprosy detection and treatment. Our results also emphasize the need for further studies addressing the impact of pragmatic health interventions aiming at controlling and eliminating the disease.

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ABSTRACT Poverty, malnutrition and neglected tropical diseases such as soil-transmitted helminthiases (STHs) interact in a multi-causal feedback network. This study aimed to assess the relationships between STHs, income and nutritional status of children in impoverished communities in the city of Caxias, Maranhao State, Northeastern Brazil. A cross-sectional survey (n=259 children) was carried out with the collection of fecal samples and assessment of sociodemographic, anthropometric, dietary and sanitation data. Hookworm infection and ascariasis presented prevalence rates of 14.3% and 9.3%, respectively. The logistic regression analysis showed that hookworm infection was more frequent in males (odds ratio [OR]=3.43; 95% confidence interval [CI]=1.45-8.08), children aged 11-15 years old (OR=3.72; 95% CI=1.19-11.62), children living in poor families (OR=2.44; 95% CI=1.04-5.68) and those living in rented houses (OR=5.74; 95%CI=1.91-17.25). Concerning ascariasis, living in the Caldeiroes community (OR=0.01; 95%CI=0-0.17) and belonging to the 11-15 years age group (OR=0.21; 95%CI=0.04-1.02) were protection factors. Poor children have a significantly lower frequency of consumption of meat, milk, vegetables, tubers and fruits than not poor children. The frequent consumption of meat, milk and tubers was associated with significant higher values in the parameter height-for-age, whereas the consumption of meat and milk positively influenced the weight-for-age. The frequencies of stunting, underweight and wasting were 8.1%, 4.9% and 2.9%, respectively. The multivariate model demonstrated that stunting was significantly associated with economic poverty (OR=2.82; 95%CI=1.03-7.70) and low weight was associated with male sex (OR=6.43; 95% CI=1.35-30.68). In conclusion, the study describes the interactions between the dimensions of development represented by income, STHs and nutritional status revealing the importance of raising income levels to improve the living conditions of families in impoverished communities in Northeastern Brazil.

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ABSTRACT Chagas disease is an important endemic morbidity in Latin America affecting millions of people in the American continent. It is caused by the protozoan Trypanosoma cruzi, and transmitted through the feces of the insect vector belonging to the subfamily Triatominae. The present conducted an entomological survey of triatomines and analyzed entomological indicators, such as the rate of infestation, colonization, triatomine density and natural infection in rural communities in the municipality of Campinas do Piaui, Piaui State, in the Northeast region of Brazil. Data on the search of triatomines performed in 167 domiciliary units (DUs), harvested during the period of February to July 2019, in 12 rural communities were analyzed. The capture of triatomines occurred in all studied communities, being 76 the number of positive DUs, of the 167 surveyed, presenting a global rate of infestation of 45.51%. Two triatomines species were collected: Triatoma brasiliensis (98.49%) and T. pseudomaculata (1.51%), the first was found in the domiciliary and peridomiciliary areas, while the second was captured only in peridomiciliary areas. The index of colonization was 17.1%. Natural infection was observed only in 5.44% of T. brasiliensis samples. The entomological survey was conducted in rural communities, showingthe risk of transmission of Chagas disease to the local population, requiring continuous entomological surveillance and vector control.

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ABSTRACT Meningoencephalitis is a syndrome of multiple etiologies associated with important morbidity and mortality. It may be caused by various infectious agents (viruses, bacteria, parasites and fungi). Establishing the etiology of meningoencephalitis is crucial for early and specific treatment. Molecular assays such as the multiplex polymerase chain reaction (PCR) offer an alternative in diagnosing central nervous system infections. This study aimed to describe the performance of an automated multiplex molecular test from patients with suspected meningitis and meningoencephalitis in a tertiary referral complex in Medellin, Colombia. Thus, a prospective study was performed in 638 cerebrospinal fluid samples from January 2017 to July 2019. Molecular detections were carried out by means of the FilmArray® Meningitis/Encephalitis (M/E) Panel from bioMérieux, France, and by conventional tests. Univariate analyses for microbiological and demographic characteristics were performed. Accuracy of the bacterial/fungal PCR assay compared to cultures was also performed. Among patients, 57.7% were male, the median age was 24 (IQR: 6 - 47) years old. The overall positivity was 15.2% (97 detections) and viruses were detected in 45.5% of the samples, bacteria in 43.5% and fungi in 10.8%. The most frequent etiological agents were: Streptococcus pneumoniae (16%), Cryptococcus neoformans/gatti (11.3%) and Herpes simplex virus (10.3%). Four double detections were found. Almost half of positive detections were in patients under 15 years old. This molecular approach is reliable and easily implantable into a laboratory routine, increasing the capacity of detection of bacterial and viral causative agents of meningitis, possibly playing a relevant role in the clinical context.

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ABSTRACT Leishmaniasis comprises a group of zoonotic diseases caused by protozoa belonging to the Leishmania genus, noting that the visceral form is the most severe and lethal, if untreated. Nowadays visceral leishmaniasis is widespread in Brazil and the Adamantina microregion, located in the west of Sao Paulo State, has been affected by Human American Visceral Leishmaniasis (HAVL) since 2004. We evaluated the epidemiological profile of HAVL in the Adamantina microregion through a Geographic Information Systems (GIS) and established its incidence rate by location and time. Notified cases were provided by the Sao Paulo State Epidemiological Surveillance Center. Home addresses of patients who tested positive to HAVL were converted into geographic coordinates through the Google Geocoding Application Programming Interface submitted to ArcMap 10.5 System for georeferencing. Kernel spatial analyses were performed to obtain the incidence distribution and the total area involvement rate. From 2004 to 2018, 325 cases of HAVL were diagnosed in 11 of the 12 municipalities belonging to the of Adamantina microregion. The disease has disseminated to the Northwest and East-Southeast directions, taking place along the Comandante Joao Ribeiro de Barros highway, with higher incidences rates in the municipalities where the highway passes. HAVL incidence was higher in children aged between 0 to 9 years and in the elderly; there was no difference in relation to sex and the majority of cases were located in urban areas. The determination of the epidemiological profile and the the spread of disease patterns can indicate possible areas of vulnerability, in order to contribute to the management and prevention of the disease through a strategic resources optimization.

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ABSTRACT The aim of this study was to evaluate the concordance between two versions of the scoring system (2011 and 2019), recommended by the Brazilian Ministry of Health, for the diagnosis of pulmonary tuberculosis (PTB) in children and adolescents. A retrospective descriptive study was performed to assess the medical records of children and adolescents with PTB, in TB units from Brazilian cities located in Rio de Janeiro, Minas Gerais, and Parana States, from January 1 st , 2004, to December 1 st , 2018. Patients aged 0 to 18 years old with a diagnosis of PTB were included. The comparison between the two scoring systems showed a moderate concordance according to the κ coefficient value = 0.625. Fourteen patients showed a reduction in the TB score, going from 30 points in the 2011, to 25 points or less in the 2019 one. Seventy one percent of these 14 patients had radiological changes suggestive of PTB and 86% had tuberculin skin tests greater than 10 mm. The study concluded that a moderate agreement was observed between the 2011 and 2019 scoring systems, with an increase in the number of patients scoring 25 points or less in 2019, which can eventually hinder the diagnosis of PTB.

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ABSTRACT Syphilis is a public health problem, especially in pregnant women, due to the risk of transmission to the fetus and the involvement of the central nervous system, causing neurosyphilis. A case-control study was carried out to analyze the variables associated with neurosyphilis in Brazilian newborns of pregnant women with syphilis admitted for childbirth. Newborns were submitted to treponemal and non-treponemal tests, cerebrospinal fluid analysis, and long bone radiography. Newborns diagnosed with neurosyphilis and congenital syphilis were defined as cases and controls, respectively. The length of hospitalization and mean cost of neurosyphilis treatment were also evaluated. Twenty-one cases of newborns with neurosyphilis and 42 controls with congenital syphilis were included in the study. Out of 63 pregnant women with syphilis, 95.2% (60/63) received prenatal care, 74.6% (47/63) were diagnosed with syphilis during this period, 31.9% (15/47) underwent treponemic tests, 80.8% (38/47) were treated with penicillin and only 46.8% (22/47) of the partners received the treatment. Clinical complications such as low birth weight were observed in 12.7% (8/63) of the newborns. About 50.8% (32/63) of the newborns were hospitalized due to syphilis complications and each case of neurosyphilis spent at least US$ 881.48 on treatment and hospitalization. The results showed that the prenatal coverage is not sufficient to prevent neurosyphilis. The late diagnosis of syphilis in pregnant women and inadequate follow-up of sexual partners may favor the vertical transmission of T. pallidum in pregnant Brazilian women. Thus, improving the quality of health services is important for a more effective control of neurosyphilis.

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ABSTRACT This study aimed to investigate a methodology for discriminating viable and non-viable T. gondii oocysts in water. Analyses included two steps: (i) microscopic investigation with vital dyes; (ii) molecular investigation, using a real time PCR (qPCR), after parasite treatment (or not) with propidium monoazide (PMA). The method was called qPCR-PMA. Oocyst aliquots were incubated (15 min) at 25 ºC or 100 ºC and analyzed by microscopy, after trypan blue and neutral red staining. Microscopic investigation determined viable and non-viable oocysts. For the molecular investigation, both aliquots of oocysts were treated with PMA. Non-viable oocysts, after PMA treatment, exhibited an inhibition of DNA amplification by qPCR. Although analyses were carried out with oocysts treated experimentally, these results suggest that qPCR-PMA can be a useful strategy to distinguish viable and non-viable T. gondii oocysts in water safety testing, showing if water is safe to drink.

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ABSTRACT Leptospirosis is an acute infection caused by pathogenic species of the genus Leptospira, which affects humans and animals in all world. In severe forms of the disease, kidneys, liver and lungs are the main affected organs, resulting in acute kidney injury, jaundice and pulmonary hemorrhage. Previous post-mortem studies have shown that lesions are not limited to these organs. Cardiac and striated muscle injuries have already been reported, but the pathophysiology of cardiac and skeletal lesions in leptospirosis is not fully understood. It has been suggested that the tissue damage observed in leptospirosis could be directly mediated by leptospires or by their toxic cellular components. LipL32 and Lp25 are leptospira membrane proteins with unknown functions, that are present only in pathogenic strains of Leptospira spp. Both proteins induce skeletal muscle lesions similar to those observed when normal guinea pigs are inoculated with leptospires. Through immunohistochemistry, this study showed the presence of LipL32 and Lp25 proteins on muscle cell membranes and in the underlying cytoplasm of skeletal muscles, as well as focal lesions in cardiac tissues of fatal cases of leptospirosis. Altogether, these results reinforce that both proteins can be important factors in the pathogenesis of leptospirosis.

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ABSTRACT Acinetobacter baumannii is one of the most frequent nosocomial pathogen capable of acquiring resistance to different antimicrobials. The aim of this study was to investigate the activity of tetracycline, doxycycline and minocycline, the prevalence of tet(A) and tet(B) determinants, and the role of efflux pump in tetracycline resistance among the A. baumannii clinical isolates. Susceptibility of 98 A. baumannii isolates to tetracyclines was evaluated by disk diffusion method. The presence of active efflux pump was investigated by determination of the minimum inhibitory concentration (MIC) of tetracycline using the carbonyl cyanide 3-chlorophenylhydrazone (CCCP). Polymerase chain reaction (PCR) was performed to investigate the presence of tet(A) and tet(B) determinants in tetracycline-resistant isolates. The rate of resistance to tetracycline, doxycycline and minocycline was 47.95%, 0%, and 30.61%, respectively. Among the 47 tetracycline-resistant isolates, 29.79% were originated from burned patients and showed MIC ranging from 128-256 μg/mL with both MIC 50 and MIC90 values of 256 μg/mL, while 70.21% were from ventilator-associated pneumonia (VAP) patients and had MIC values ranging from 32-1024 μg/mL, with MIC50 and MIC90 of 512 μg/mL and 1024 μg/mL, respectively. The tet(B) gene was found in 61.7% of tetracycline-resistant isolates, while none of the isolates carried the tet(A) gene. CCCP led to 2-128-fold reduction in tetracycline MIC of the tested isolates. The results showed that doxycycline and minocycline are promising agents for the treatment of A. baumannii infections. This study has also revealed the role of efflux activity in the resistance to tetracycline of A. baumannii isolates. The emergence of resistance to these agents is likely due to the spread of clones presenting with a higher prevalence of resistance determinants.

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ABSTRACT Coronavirus disease 19 (COVID-19) is caused by SARS-Cov-2 and the manifestations of this infection range from an absence of symptoms all the way up to severe disease leading to death. To estimate the prevalence of past infection in a population, the most readily available method is the detection of antibodies against the virus. This study has investigated the prevalence of anti-SARS-CoV-2 antibodies in outpatients of the Hospital das Clinicas, in Sao Paulo city (Brazil), which is a large university hospital belonging to the public health system that cares for patients with complex diseases who need tertiary or quaternary medical care. Our serological inquiry was carried out for 6 weeks, with once-a-week blood sampling and included 439 patients from several outpatient services. Overall, 61 patients tested positive for anti-SARS-CoV-2 IgG (13.9%); 56.1 % of the patients live in Sao Paulo city, with the remaining living in other towns of the metropolitan area; 32.8% of the patients testing positive for IgG antibodies to SARS-CoV-2 were asymptomatic, 55.7% developed mild or moderate disease and 11.5% had to be hospitalized. The prevalence of SARS-CoV-2 positive serology was lower among patients who had received the seasonal influenza vaccine compared to the ones who did not. These findings may indicate that those individuals care more about health issues, and/or that they have a better access to health care and/or a better quality of health care service. The large proportion of patients who were unaware of having had contact with SARS-CoV-2 deserves attention, reflecting the scarcity of tests performed in the population.

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ABSTRACT This study has estimated the risk of Leishmania transmission via blood transfusion in one of the largest blood banks in Northeastern Brazil, where visceral leishmaniasis is endemic. Five hundred blood samples from donors were tested for circulating Leishmania spp. DNA by real-time PCR. Positive samples were tested by a species-specific conventional PCR targeting Leishmania infantum . Overall, 6.2% (95% CI: 4.1–8.3%) of the samples carried Leishmania DNA and in one sample the species was confirmed as L. infantum . No statistically significant differences were found in relation to gender, sex, education level, incomeas well as the place of residence between positive and negative blood donors. Our results confirm the presence of asymptomatic Leishmania carriers among blood donors in a large blood bank in Northeastern Brazil. Considering the studied population, we estimate that for every 1,000 blood donors screened, 41 to 83 will be positive for Leishmania DNA. This finding reinforces the urgent need for elaborating specific Blood bank guidelines to allow the early detection of asymptomatic Leishmania carriers among blood donors before their blood products are transfused to uninfected individuals.

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ABSTRACT Leprosy is a public health problem due to the physical disabilities and deformities it causes. This study aimed to describe new leprosy cases using an operational classification and analyzing spatial patterns by means of epidemiological and quality indicators of health services in Pernambuco State, Brazil, between 2005 and 2014. This was an ecological study performed in 184 municipalities grouped into 12 health regions units for analysis. To analyze spatial patterns, the Bayesian local empirical method and Moran's spatial autocorrelation indicator were applied and box and Moran maps were used. Individuals aged ≥15 years old, grade zero physical disability and complete remission as the treatment outcome were predominant in both paucibacillary and multibacillary cases, the only difference was the predominance of females (n=9,286; 63.00%) and males (n=8,564; 60.70%), respectively. These variables were correlated (p<0.05) with the operational classification. The overall detection rate showed three high-priority areas; the indicator rate of grade 2 physical disability revealed clusters in regions IV, V, and VI; and the indicator rate of cases with some degree of disability showed precarious municipalities in seven health regions. Pernambuco maintains an active chain of transmission and ongoing endemicity of leprosy. Therefore, spatial analysis methods allow the identification of priority areas for intervention, thereby supporting the disease elimination strategy.

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ABSTRACT The circulation of arboviruses throughout the world and the maintenance of lymphatic filariasis endemicity in tropical countries, combined with the lack of vaccines and specific treatments, highlight the importance of reducing the populations of mosquitoes involved in the transmission of these pathogens, Aedes aegypti and Culex quinquefasciatus. To contribute to the development of new strategies for monitoring and controlling these culicids, we evaluated the performance of the Double BR-OVT trap individually and in pairs, in the field. After 18 months, the Double BR-OVT traps captured a mean of 3.5 ± 7.4 and 1.8 ± 3.2 of Culex and Aedes /residence/cycle, respectively, in addition to 410 ± 588.3 Aedes eggs/residence/cycle. When installed in pairs, the Double BR-OVT traps collected three times more adult mosquitoes of C. quinquefasciatus (9.4 ± 8.3 Culex /residence/bimester) and two times more Aedes spp. (3 ± 3.2 Aedes /residence/bimester) in comparison with the traps installed individually (2.6 ± 7.1 and 1.5 ± 3.2 Culex and Aedes /residence/bimester, respectively) (p < 0.05). The Double BR-OVT trap has an exceptional advantage: it aggregates different functionalities into a single instrument, as this type of trap can concomitantly collect eggs and adult mosquitoes of C. quinquefasciatus and A. aegypti , a feature that makes it a potentially useful tool among the strategies for monitoring and controlling these mosquitoes.

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ABSTRACT Stenotrophomonas maltophilia has emerged as an important opportunistic pathogen in the last decade. Increased resistance to sulfamethoxazole/trimethoprim (SMX/TMP) has been reported in S. maltophilia strains in the past few years, leading to few therapeutic options. We conducted a prospective multicenter study at two Brazilian teaching hospitals that identified S. maltophilia isolates and evaluated their antimicrobial susceptibility profile, SMX/TMP resistance genes and their clonality profile. A total of 106 non-repeated clinical samples of S. maltophilia were evaluated. Resistance to SMX/TMP was identified in 21.6% of the samples, and previous use of SMX/TMP occurred in 19 (82.6%). PCR detected the sul1 gene in 14 of 106 strains (13.2%). Of these isolates, nine displayed resistance to SMX/TMP. The resistant strains presented a polyclonal profile. This opportunistic pathogen has emerged in immunocompromised hosts, with few therapeutic options, which is aggravated by the description of emerging resistance mechanisms, although with a polyclonal distribution profile.

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ABSTRACT Surveillance of Rotavirus A (RVA) throughout the national territory is important to establish a more complete epidemiological-molecular scenario of this virus circulation in Brazil. The aim of the present study was to investigate the genetic diversity of RVA strains circulating in Tocantins State (Northern Brazil) during six years of post-vaccination follow-up (2010-2016). A total of 248 stool samples were screened by next generation sequencing and 107 (43.1%) nearly full length RVA genome sequences were obtained; one sample was co-infected with two RVA strains (G2/G8P[4]). Six G and P genotypes combinations were detected: G12P[8] strains (78.6%), as well as the G3P[8] (9.3%) and G1P[8] (0.9%) were associated with a Wa-like genogroup backbone. All G2P[4] (5.6%) and G8P[4] (2.8%) strains, including the mixed G2/G8P[4] infection (0.9%) showed the DS-1-like genetic background. The two G12P[4] strains (1.9%) were associated with distinct genetic backbones: Wa-like and DS-1-like. The phylogenetic analysis revealed the circulation of lineages G1-I, G2-IV, G3-III, G8-I and G12-III, and P[4]-V and P[8]-III of the VP7 and VP4 genes, respectively. Conserved clustering pattern and low genetic diversity were observed regarding VP1-VP3 and VP6, as well as NSP1-5 segments. We identified the same RVA circulation pattern reported in other Brazilian regions in the period of 2010-2016, suggesting that rural and low-income areas may not have a different RVA genotypic distribution compared to other parts of the country. The unique presentation of whole-genome data of RVA strains detected in the Tocantins State provides a baseline for monitoring variations in the genetic composition of RVA in this area.

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ABSTRACT Tropical diseases are mainly found in the tropical regions of Asia, Africa and Latin America. They are a major Public Health problem in these regions, most of them are considered neglected diseases and remain as important contributors to the development of AKI (Acute Kidney Injury), which is associated with increased patients’ morbidity and mortality. In most countries, kidney disease associated to tropical diseases is attended at health services with poor infrastructure and inadequate preventive measures. The long-term impacts of these infections on kidney tissue may be a main cause of future kidney disease in these patients. Therefore, the investigation of novel kidney injury biomarkers in these tropical diseases is of utmost importance to explain the mechanisms of kidney injury, to improve their diagnosis and prognosis, as well as the assessment to health systems by these patients. Since 2011, our group has been studying renal biomarkers in visceral and cutaneous leishmaniasis, schistosomiasis, leptospirosis and leprosy. This study has increased the knowledge on the pathophysiology of kidney disease in the presence of these infections and has contributed to the early diagnosis of kidney injury, pointing to glomerular, endothelial and inflammatory involvement as the main causes of the mechanisms leading to nephropathy and clinical complications. Future perspectives comprise establishing long-term cohort groups to assess the development of kidney disease and the patients’ survival, as well as the use of new biomarkers such as urinary exosomes to detect risk groups and to understand the progression of kidney injuries.

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ABSTRACT This review provides a general overview on the positivity and persistence of Zika virus (ZIKV) in female genital tract (FGT) of non-pregnant women and animals, as well as in cell cultures, and its influence on FGT health. We performed a systematic review based on the PRISMA statement to identify studies focused on “Zika virus” and “non-pregnant female” in PubMed, Embase, Scopus Scholar and Web of Knowledge databases of full-text papers and abstracts published in English, with no restrictions regarding the initial date of publication, up to August 2019. Our search terms yielded 625 records, that were 108 after removal of duplicates, leaving 517 items for title and abstract reviews. Of these, 475 did not meet the inclusion criteria, leaving 42 records for full-text review and resulting in the exclusion of 6 additional records. The remaining 36 met our inclusion criteria. Variations were observed regarding the presence and persistence of ZIKV in lower and upper genital samples. However, the FGT was the place in which ZIKV RNA has been detected, sometimes for relatively long periods, even after the clearance from blood and urine. In addition to the vagina and cervix, the endometrium, uterus and ovary (oocytes and follicles) could also be involved in persistent ZIKV infections. Further prospective studies are needed to assess the effect of ZIKV on FGT health.

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ABSTRACT Leptospirosis is a reemerging zoonosis caused by bacteria of the genus Leptospira sp. with global importance in the medical and veterinary fields, being responsible for about 59 thousand deaths each year in the world. The use of Geographic Information Systems (GIS) in the health sector is propitious and has been adopted by human and animal health professionals as an important tool in spatial analyses of health. The objective of this study was to conduct a systematic review on the geoprocessing and spatial analysis techniques adopted for mapping risk areas of human and animal leptospirosis. The articles were collected on scientific platforms by entering the following terms: SIG/GIS, leptospirose/leptospirosis, area de risco/risk area and distribuicao espacial/spatial distribution, and included in the study if they met the following criteria: a) publication in the period from 1998 to 2017; b) identification of risk areas and/or spatial distribution of leptospirosis as one of the research topics; and c) application of GIS in the methodology. As a result, we found 40 articles, published by 15 different countries, which adopted GIS for the spatial analysis and identification of risk areas of leptospirosis. Among these, only 45% (18) conducted an spatial statistical analysis. Brazil and USA had the highest numbers of publications, 16 and 7 articles, respectively. From 2007, the use of GIS and spatial analysis techniques, applied to the theme of this study, have been intensified and diversified, and 93% of the articles elected for this review were published from 2007 to 2017. The results point to a progressive interest of health professionals in applying these techniques for monitoring and conducting epidemiological analyses of leptospirosis, besides indicating a greater need for intersectoral integration between health professionals and others, in the use of spatial analysis and GIS techniques.

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ABSTRACT Coronavirus disease 2019 (COVID-19) is an infectious disease initially reported in China and currently worldwide dispersed caused by a new coronavirus (SARS-CoV-2 or 2019-nCoV) affecting more than seven million people around the world causing more than 400 thousand deaths (on June 8th, 2020). The diagnosis of COVID-19 is based on the clinical and epidemiological history of the patient. However, the gold standard for COVID-19 diagnosis is the viral detection through the amplification of nucleic acids. Although the quantitative Reverse-Transcription Polymerase Chain Reaction (RT-PCR) has been described as the gold standard for diagnosing COVID-19, there are several difficulties involving its use. Here we comment on RT-PCR and describe alternative tests developed for the diagnosis of COVID-19.

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ABSTRACT The present study aimed to review the existing literature and to evaluate the best dose regimen for benznidazole in adult patients with Chagas disease in the chronic phase. A systematic review was conducted followed by meta-analysis. Searches were performed in four databases, to include studies published until May 2019. The descriptors used were: “Chagas disease”, “benznidazole”, “Drug Therapy”, “Pharmacokinetics”, “Dose-response relationship, drug” and “Chronic disease”. The meta-analysis compared studies using the standard dose of 5 mg/kg/day for 30 or 60 days. A total of 608 articles were found, 23 of which were considered eligible for this review and nine were included in the meta-analysis. The studies selected and analyzed were published between 1996 and 2018, with various benznidazole dose regimens, ranging from 2.5 mg/kg/day to 10 mg/kg/day, for 30 to 80 days of treatment. The results pointed to a great diversity of dose regimens, thus there is no consensus on the optimal dose regimen for benznidazole in the chronic phase of Chagas disease.

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ABSTRACT The consequences of COVID-19 infecting pregnant women and the potential risks of vertical transmission have become a major issue. Since little is currently known about COVID-19 in pregnancy, the understanding of COVID-19 in this particular group will be updated in time, and a comprehensive review will be useful to evaluate the impact of COVID-19 in pregnancy. Based on recently published literature and official documents, this review provides an introduction to the pathogenesis, pathology, and clinical features of COVID-19 and has focused on the current researches on clinical features, pregnancy outcomes and placental histopathological analysis from pregnant women infected with SARS-CoV-2 in comparison with SARS-CoV and MERS-CoV. These viruses trigger a cytokine storm in the body, produce a series of immune responses, and cause changes in peripheral leukocytes and immune system cells leading to pregnancy complications that may be associated with viral infections. The expression of ACE2 receptors in the vascular endothelium may explain the histological changes of placentas from pregnant women infected by SARS-CoV-2. Pregnant women with COVID-19 pneumonia show similar clinical characteristics compared with non-pregnant counterparts. Although there is no unequivocal evidence to support the fetal infection by intrauterine vertical transmission of SARS, MERS and SARS-CoV-2 so far, more and more articles began to report maternal deaths due to COVID-19. In particular, from February 26, 2020 (date of the first COVID-19 case reported in Brazil) until June 18, 2020, Brazil reported 124 maternal deaths. Therefore, pregnant women and neonates require special attention regarding the prevention, diagnosis and management of COVID-19.

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ABSTRACT This narrative review summarizes the main aspects underlying the new coronavirus SARS-CoV-2, its epidemiology, pathophysiology, pointing to differences of SARS-CoV-2 main receptors ACE2, in terms of expression and the amount of soluble ACE2 in the circulation of children, men and women, and also in those with risk factors such as the smokers and pregnant women or presenting with comorbidities (diabetes, obesity, hypertension and other cardiovascular diseases, renal and CNS pre-existing diseases). Clinical manifestations in adults and children were also described, emphasizing the particularities already seen in children, regarding signs, symptoms, viral excretion time and the involvement of all organs and systems. The COVID-19 in the pediatric population was divided into two sections: one dedicated to previously healthy children and adolescents with COVID-19, and the other to those who live with comorbidities and acquired COVID-19. A few paragraphs were reserved to the recently described severe multisystemic inflammatory syndrome associated with COVID-19 (MIS-C) that shares certain characteristics with Kawasaki disease. Some studies on the infection in pregnant and postpartum women, as well as neonates were shown. This review has also covered the laboratory diagnosis of COVID-19, passing through the imaging diagnosis made by the chest tomography revealing ground glass patching opacities, and results of non-specific exams such as the total blood with lymphopenia, the coagulation tests with increased prothrombin times, as well as marked increments of the D-dimer, troponin and proinflammatory cytokines. In the section devoted to the specific laboratory diagnosis of COVID-19, the most used RT-PCR protocols were described and some studies on the serological diagnosis with IgA, IgM and IgG detection were detailed, including the use of rapid immunochromatographic assays and discussing the ideal period after the onset of symptoms to perform each type of test. In the end, the management of pediatric patients with COVID-19 based mainly on supportive measures has been briefly commented.

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ABSTRACT Rabies encephalitis is a fatal zoonotic viral disease transmitted to humans either by domestic animals like dogs and cats or by wild animals like bats, skunks and raccoons. We present the case of a 25-year-old woman admitted due to behavioral disorders, generalized paresthesiasand acute respiratory deterioration compatible with a respiratory distress syndrome (ARDS) requiring orotracheal intubation, mechanical ventilation and empirical initiation of antibiotic and antiviral therapy. Chest tomography showed pneumomediastinum and changes suggestive of pulmonary infection. In the presence of neurological symptoms, a central nervous system (CNS) infection was suspected and the cerebrospinal fluid showed no pleocytosis, hiperproteinorrachy without glucose consumption; cranial CT scan was normal. During hospitalization, the family reported that the patient was bitten by a cat 30-day prior to the onset of rabies-like symptoms, and the animal was sacrificed. The patient had an unfavorable clinical evolution, with electroencephalographic activity dissociation evidenced by video telemetry. The zoonotic exposure led to the suspected diagnosis of rabies infection. The patient died and the suspected diagnosis was confirmed by histopathology, with presence of Negri bodies on cerebellum Purkinje cells and a positive immunofluorescence test for rabies virus. Both, initial extra-neural manifestations and late reporting of rabies exposure led to delayed diagnosis.

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ABSTRACT A 37-year-old male patient, Fitzpatrick skin phototype IV, a student living in Belem, Amazon region, in 2015 had a confirmed diagnosis of acquired immunodeficiency virus (HIV) infection, but did not initiate antiretroviral treatment at his own option. Three years after the diagnosis, erythematous maculae appeared on the dorsum of the nose with rapid progression to the entire face, with posterior diffuse infiltration and appearance of nodules on the chin and shoulder. In December 2018, the patient presented with exacerbation of the condition with an increase in infiltrated violaceous plaques and disseminated violaceous nodules. A histopathological biopsy of the skin was performed, confirming the diagnosis of angiomatoid proliferation suggestive of Kaposi’s sarcoma (KS), with an important dissemination of this disease to the noble organs. In addition, it is important to note that he only started antiretroviral therapy (ART) after the exacerbation of Kaposi (December 2018). In such cases, chemotherapy associated with ART is crucial for the treatment and follow-up of the patient, since Kaposi’s sarcoma develops relatively low in patients who do not have immunodeficiency.

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ABSTRACT Cryptococcosis is an opportunistic fungal infection causes significant disease predominantly in immunocompromised patients. Here we present an excepcional case of disseminated cryptococcosis with pulmonary and cerebral involvement in an immunocompetent patient with no apparent predisposing factors at the time of hospital admission. We described a case of an apparently immunocompetent 66-years old man admitted to hospital with a one-month history of cough, fever and vertigo. During hospitalization, thorax imaging was suggestive of lung metastasis, therefore, he went through several investigations. During hospitalization, he developed neurological symptoms and subsequently underwent a lumbar puncture. Cerebrospinal fluid (CSF) culture was positive for Cryptococcus spp. isolated on Sabouraud’s dextrose agar and bird seed agar. In addition, the direct microscopy examination was positive for the India ink test, as well as with the latex agglutination test for cryptococcal polysaccharide antigen (CrAg) in CSF, while serum CrAg was negative. Despite the absence of classic immunocompromising features, he was treated with amphotericin B and fluconazole due to suspected disseminated cryptococcal infection. Later, he was diagnosed with prostatic adenocarcinoma. Upon successful completion of treatment for disseminated cryptococcosis, the patient underwent radical prostate ablation surgery as a treatment forprostatic adenocarcinoma. This exceptional case emphasizes the high degree of suspicion of atypical infections, and in these cases, it is particularly important to consider fungal infections in hitherto healthy patients with no apparent predisposing factors. Although Cryptococcus spp. is predominantly reported in patients with hematological malignancies, cryptococcosis investigation should also be considered as part of the initial workup of patients with a new diagnosis of a solid tumour prior to chemotherapy or radiotherapy.

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ABSTRACT In this report, we described a rare case of Mycobacterium abscessus subsp. bolletii keratitis in a young healthy male, in the absence of risk factors.

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ABSTRACT It is essential for health care providers to be familiared with the full spectrum of clinical presentations of syphilis. We present herein a case of syphilide psoriasiforme, an uncommon but well recognized clinical presentation of secondary syphilis. A 46-year-old HIV-infected female patient was referred to our attention with a presumptive diagnosis of palmoplantar psoriasis. On examination, there were exuberant pinkish-red papules and plaques covered with a thick silvery scale in the palms, flexor surfaces of the wrists, and the medial longitudinal arches of the feet. Serological and histopathological analyses uncovered the diagnosis of syphilis. Clinical remission was obtained after treatment. A detailed review of the literature on syphilide psoriasiforme, including descriptions from older syphilology textsis provided. The present case report emphasizes the need for clinicians to have a heightened awareness of the varied and unusual clinical phenotypes of syphilis.

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ABSTRACT Coccidioidomycosis is an endemic disease of arid regions in the Western hemisphere. Its clinical presentation varies from asymptomatic nodules on chest x-rays to disseminated disease. We present the case of a 48-year-old man with a hard and heterogeneous tumor in the posterior aspect of the right testis. Color flow doppler testicular ultrasonography was performed and two nodular masses in the tail of the right epididymis were identified. An epididymectomy was performed and histopathological examination revealed coccidioidomycosis. After diagnosis, the patient was successfully treated with fluconazol.

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ABSTRACT In Latin America, the causative agent of kala-azar is the intracellular protozoan Leishmania infantum. Most cases in South America are reported in Brazil. Worldwide, it mainly affects Bangladesh, Ethiopia, India, South Sudan and Sudan. Despite the high morbidity and lethality of kala-azar, most infections are asymptomatic. However, a small portion of patients evolves with recurrence of kala-azar becoming symptomatic even after all available drug treatments. Kala-azar is not a formal indication for splenectomy in adults. Splenectomy is recommended as a saving measure, when kala-azar is associated with symptomatic hypersplenism and for drug-resistant cases. In the study, we report two cases of kala-azar with splenomegaly that presented several hospitalizations due to the recurrence of the kala-azar, in addition to hospitalizations for normalizing the blood count. After splenectomy, kala-azar cases and the effects of hypersplenism are cured. Thus, splenectomy should be seen as a surgical treatment option with a curative purpose in patients with recurrent kala-azar, in whom the possibilities of drug therapy have been exhausted and even so they progressed with hypersplenism and clinical repercussions.

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ABSTRACT This is a fatal case of fungaemia due to Cryptococcus albidus in an elderly woman with type 2 diabetes mellitus who presented pleural effusion as the only clinical presentation. Not only infections of the pleura are uncommon presentations of pulmonary cryptococcosis, but these infections due to non-C. neoformans species are extremely rare. This report places C. albidus on the growing number of disseminated mycosis-causing agents in diabetic patients.

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ABSTRACT Chikungunya, a viral infection that presents with fever, rash and polyarthritis, is usually an acute febrile illness. Uncommon neurological manifestations include meningoencephalitis, encephalitis, myelitis, Guillain-Barre syndrome, myelopathy and neuropathy. During an outbreak of the disease in La Reunion Island, abnormalities were observed in the magnetic resonance imaging (MRI) of patients with encephalitis and acute disseminated encephalomyelitis, showing bilateral, frontoparietal, white matter lesions with restricted diffusion, similar to our case. We report a 57-year-old male patient with comorbidities, admitted with high fever, arthralgia, asthenia, vomiting, psychomotor agitation, behavioral changes and seizures. Cerebrospinal fluid (CSF) values revealed pleocytosis (98 cells/mm3 with 68% lymphocytes and 12% monocytes) and high levels of protein (161 mg%). Brain MRI showed hyperintense lesions in the temporal and frontal lobes and bilaterally in the posterior thalamus. CSF serology was positive for IgM antibodies to Chikungunya virus. Encephalitis due to an acute viral infection by Chikungunya was diagnosed. The patient’s clinical condition worsened and he died on the twenty-fourth day of admission to our hospital.

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ABSTRACT We report the case of a patient with cutaneous leishmaniasis who showed a rapidly progressing ulcerative lesion after traveling to multiple countries where different Leishmania species are endemic. Diagnosis of Leishmania tropica, an exotic species in Mexico was established by using serological and molecular tools.

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ABSTRACT Dermatophytes are known as a common cause of superficial mycosis, but atypical presentations in immunosuppressed patients make the diagnosis more challenging. Here, we report a case of a 39-year-old patient, a renal transplant recipient from a living donor, who presented with atypical cutaneous lesions of lower extremities caused by Arthroderma gypseum (Nannizzia gypsea), four months after receiving a renal transplant. It is important to highlight the importance of the early detection of fungal infections in immunosuppressed patients. Clinicians should have a high degree of suspicion for the early detection and treatment of the cases.

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ABSTRACT Myiasis represents a group of neglected tropical diseases caused by the infestation of vertebrate tissues by dipterous larvae. We herein report an imported case of foruncular myasis caused by Dermatobia hominis in Mexico City. The species was confirmed by DNA sequencing and phylogenetic reconstruction analysis.

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ABSTRACT Capnocytophaga is a group of facultative anaerobic gram-negative bacteria present in the oral cavity of humans, dogs and cats, as part of their normal oral flora. Here, we described two cases of bloodstream infections (BSI) caused by Capnocytophaga in neutropenic autologous hematopoietic stem cell transplantation (auto-HSCT) patients with mucositis (Grade I and Grade III) identified by Maldi-Tof. They were successfully treated with β-lactam (meropenem and piperacillin-tazobactam). The species C. sputigena was confirmed by 16S rRNA gene sequencing in one patient. The review of literature showed that C. ochraceae was the most frequent species causing BSI in auto-HSCT patients and that the patients usually presented mucositis and were neutropenic at the onset of the infection.

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ABSTRACT There are few data on the impact of COVID-19 in pregnancy, however, analyzing these data is important to guide the clinical practice, covering the early prevention, detection, patients’ isolation, epidemiological investigation, diagnosis and early treatment. This is a report of three cases of COVID-19 confirmed by real-time reverse transcription – polymerase chain reaction (RT-PCR) of nasopharyngeal secretions collected in swabs from pregnant women in the city of Vitoria, Espirito Santo State, Brazil. In the three cases, all the patients presented with fever, one had shortness of breath, one had diarrhea, two of them reported abdominal pain and two of them had cough. The three patients progressed with a severe clinical evolution of COVID-19. The permanence in the intensive care unit (ICU) was more than 10 days. Two of them recovered and one remained in the ICU with irreversible refractory shock, multiple organ failure and died. The mode of delivery was individualized and based on the obstetric indication and severity of the maternal infection, and the cesarean section was indicated in the two severe maternal COVID-19 cases that evolved favorably. These newborns were premature and tested negative for COVID-19 by RT-PCR.

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ABSTRACT Bothrops erythromelas are serpents that belong to the Viperidae family, which are the main species responsible for human snakebites in Ceara State, Northeast Brazil. Thrombotic microangiopathy (TMA) is an uncommon group of disorders characterized by microangiopathic hemolytic anemia (MAHA), thrombocytopenia and acute kidney injury (AKI), and occurrence after snakebites have been rarely reported. In this report, we described the case of a 57 year-old-man without comorbidities who was bitten by a Bothrops erythromelas on his right ankle. He presented with pain, edema and local bleeding. Symptomatology and laboratory tests were compatible with the diagnosis of TMA. He received specific antivenom and fluids replacement without any anaphylactic reaction. The conservative treatment was effective and there was no need for red blood cells transfusion or plasmapheresis. The aim of this report was to describe the first case of thrombotic microangiopathy following Bothrops erythromelas envenoming in the Northeast Brazil, providing insights about important mechanistic pathways of Bothrops snakebite-associated TMA and how to change the prognosis of the disease.

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ABSTRACT Sporotrichosis is an implantation mycosis highly prevalent in Brazil, associated with soil activities and contact with infected animals. It has numerous clinical manifestations and its disseminated cutaneous form is uncommon, found in approximately 4% of cases and usually related to immunosuppressive conditions. We report an alcoholic 56-year-old male patient with no other comorbidities, presenting with multiple cutaneous nodules and ulcers. The patient was diagnosed with disseminated cutaneous sporotrichosis based on the isolation and identification of Sporothrix spp. The patient was initially treated with amphotericin B with satisfactory results and then with itraconazole.

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ABSTRACT In late 2019, a novel coronavirus initially related to a cluster of severe pneumonia cases in China was identified. COVID-19 cases have rapidly spread to multiple countries worldwide. We present a typical laboratory confirmed case of COVID-19 pneumonia, that was hospitalized due to hypoxemia but did not require mechanical ventilation. Although initially the patient was evaluated with a favorable outcome, in the third week of the disease, the symptomatology deteriorated due to a massive hypertensive pneumothorax with no known previous risk factor. Since the first cases of COVID-19 have been described, pneumothorax was characterized as a potential, though uncommon, complication. It has been reported that diffuse alveolar injury caused by SARS-CoV-2 can cause alveolar rupture, produce air leakage and interstitial emphysema. Although uncommon, pneumothorax should be listed as a differential diagnosis for COVID-19 patients with sudden respiratory decompensation. As a life-threatening event, it requires prompt recognition and expeditious treatment.

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ABSTRACT In late 2019, the severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) emerged as the cause of a cluster of pneumonia cases in China, and the corresponding disease was designated as Coronavirus Disease 2019 (COVID-19), spreading quickly around the world resulting in a pandemic. COVID-19 is associated with a set of coagulation abnormalities that increase the risk of thromboembolic events, especially in patients with severe/critical disease. We describe a series of five cases of mild COVID-19, treated in an outpatient clinic, which, after an apparent clinical improvement, developed acute pulmonary embolism (APE) between the third and the fourth week after the onset of symptoms, when they are mostly related to acute illness disappearance. Thromboembolic events are also a potential complication of mild COVID-19 and can manifest later in the disease course. This finding raises discussion about the prevention of thromboembolic events in selected group of patients with mild COVID-19.

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ABSTRACT Tuberculosis of the skull or calvarial tuberculosis (CTB) is rare. The literature until 2019 reported less than 60 cases of CTB in childhood. The authors describe two patients with CTB associated with other manifestations of TB, such as: spine and rib injuries, peripheral adenopathy, hepatic and splenic involvement who improved with chemotherapy. The patients were a four-year-old and an eight-year-old child, whose diagnoses were confirmed by histopathological, bacteriological or molecular investigation. Both were not infected with the human immunodeficiency virus (HIV) and did not need orthopedic treatment.

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ABSTRACT Coronavirus Disease 2019 (COVID-19) has rapidly spread worldwide. Numerous studies have shown its typical and atypical CT findings. We report one COVID-19 patient who presented with a transient pneumothorax, spontaneous pneumomediastinum (SP), as well as subcutaneous emphysema during hospitalization. Chest CT andclinical findings were discussed, and a literature review is presented. The probable cause of SP in COVID-19 was alveolar damage. Once pneumothorax and SP were present, the patient should be carefully monitored to prevent respiratory deterioration, especially when lung lesions are severe.

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ABSTRACT Disseminated tuberculosis is a severe disease with high-mortality that requires early diagnosis and treatment. Intestinal tuberculosis accounts for only 2% of tuberculosis cases worldwide and is extremely rare in children. We report a case of a 4-year-old girl admitted due to disseminated tuberculosis with extensive intestinal involvement characterized by massive intestinal bleeding and hemorrhagic shock. The severity of the intestinal involvement precluded the exclusive use of oral anti-tuberculosis drugs and the patient was successfully treated with a combination of injectable and oral anti-tuberculosis agents. We discuss the importance of a regimen with injectable drugs for treating severe forms of tuberculosis in which the intestinal involvement impaired the use of oral drugs.

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ABSTRACT Lagochilascariasis is a neglected neotropical helminthiasis, responsible for human infections through the ingestion of raw or undercooked meat from wild animals infected by larvae encysted in their tissues. It is characterized by a chronic evolution with periods of remission and recurrences, affecting mainly the head and neck regions. It can progress to death by invasion of the central nervous system. It is caused by the Lagochilascaris minor nematode, first described in 1909 and mainly found in the Brazilian Amazon ecosystem, notably in Para State. In May 2010, a 15-year-old female, born in the city of Aveiro, Para State, was hospitalized in the city of Santarem, Para State, presenting with cervical and spinal abscesses and osteolytic lesions at T3-T5 level. During hospitalization, white larvae identified as L. minor were observed in a cervical subcutaneous fistula. After the surgical drainage of abscesses, albendazole 400 mg/day was started, with improvement of symptoms. The patient abandoned the follow-up and in January 2016, during the 20th week of pregnancy, she was readmitted to the Santarem Municipal Hospital with new cervical abscesses in the infra-mammary region, with numerous larvae exiting by subcutaneous left breast fistulas. The antiparasitic treatment was restarted due to possible reactivation of the infection in the spine and dissemination to the central nervous system. Ten weeks after hospital discharge, the patient had resolution of the fistulas and decreased abscesses, without gestational complications. This is the first case report of lagochilascariasis during pregnancy, as well as of involvement of mammary glands.

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ABSTRACT Paracoccidioidomycosis caused by Paracoccidioides lutzii is endemic in the Midwest of Brazil and its clinical spectrum is still little known due to the recent identification of this fungal species. A patient resident in Southeast Brazil, but who had lived for many years in the Midwest region, presented with skin injuries, chronic cough and bilateral adrenal involvement. Paracoccidioides spp. was isolated in culture from a skin lesion biopsy. This isolate was later identified as P. lutzii using gene sequencing. A favorable initial response to treatment with itraconazole was observed, but a few weeks later, the patient developed respiratory failure and worsening of lung lesions. Evaluation by computed tomography and echocardiography were suggestive of pulmonary arterial hypertension, and a bronchoscopic biopsy showed peribronchial remodeling. The patient completed the antifungal treatment but maintained the respiratory dysfunction. The reported case shows that P. lutzii can be isolated from patients in a geographic area far from the place of infection acquisition and that, as P. brasiliensis , it can cause adrenal injury and cardio-respiratory complications as a consequence of excessive necrosis and fibrosis.

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ABSTRACT Skins infections caused by Mycobacterium marinum occur only rarely. We report one case of chronic and extensive M. marinum cutaneous infection simulating chromoblastomycosis and review the pertinent literature. A 52-year-old farmer reported a 32-year chronic skin problem on his right lower limb, resulting from contact with cacti. It consisted of skin lesion presenting with dyschromic atrophic center plate and verrucous borders with hematic crusts, extending from the knee anteriorly to the inferior third of the right leg. Mycobacterium marinum infection was detected by histopathological examination of a skin fragment, culture for mycobacteria and genetic mapping of the culture material. The patient was successfully treated with Ethambutol, Rifampicin and Trimethoprim-Sulfamethoxazole. The clinical and histopathological findings of M. marinum infection is nonspecific showing clinical polymorphism and bacilli are rarely evident on histopathological examination. Given these difficulties, it is essential to perform tissue culture in a suspicious case and it is important keep this infection in mind in patients with long-lasting indolent verrucous lesions and a history of exposure to sea water, freshwater, aquaria or fish.

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ABSTRACT Hemophagocytic lymphohistiocytosis is a rare complication in Rickettsia typhi infections. We report the case of a 2-year-old boy with sudden night-onset fever, pallor, neck adenopathy and erythematous macular rash on the thorax, thighs and buttocks. During admission, he developed hyponatremia, hypoalbuminemia, anemia, thrombocytopenia, leukopenia, neutropenia, liver damage, hemorrhages and persistent fever. No hematological improvement was observed after the initial management, neoplastic diseases were discarded by bone marrow aspiration and lymph node biopsy; hemophagocytic lymphohistiocytosis was diagnosed. By immunohistochemistry and indirect immunofluorescence, murine typhus was also diagnosed and doxycycline was started with transitory recovery. Later, the child developed kidney failure and distributive shock that evolved to cardiac arrest and death. This is the first case report in Mexico on a fatal murine typhus associated with hemophagocytic lymphohistiocytosis in which the etiology was evidenced by histopathology.

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ABSTRACT Dengue is a viral disease, caused by an arbovirus of the genus Flavivirus. In Brazil, its incidence rate is high with a broad clinical spectrum. This report discusses a rare case of dengue associated with cutaneous leukocytoclastic vasculitis and pericardial effusion with eminence of cardiac tamponade in a previously healthy patient with no comorbidities. The serology for dengue was positive and the histopathological analysis of the cutaneous lesions confirmed the diagnosis of leukocytoclastic vasculitis. After receiving treatment, the patient’s condition greatly improved.

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ABSTRACT COVID-19 is a viral disease caused by SARS-CoV-2 that compromises the host immune response in severe cases, promoting a hyperinflammation that results in acute lung injury and multiple organs failure. In this context, patients presenting with immune-related diseases, such as Crohn’s disease, affected by COVID-19, may have an uncertain prognosis. We report on a case of a young female patient with a severe Crohn’s disease that presented with COVID-19 pneumonia and a favorable outcome even maintaining the use of adalimumab, TNF - alpha inhibitor and prednisone. This case raises the hypothesis that aside from prednisone, TNF-α inhibitors such as adalimumab could be used to stop the progression to COVID-19 complications by blocking the TNF-alpha-driven inflammatory process that occurs in severe COVID-19.

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ABSTRACT Candidemia is a significant cause of bloodstream infections (BSI) in nosocomial settings. The identification of species can potentially improve the quality of care and decrease human mortality. Quantitative PCR (qPCR) was evaluated for Candida albicans detection using culture suspensions containing C. albicans , spiked human blood, the cloned qPCR target fragment (ITS2 region) and the results of these assays were compared. The assays showed a good detection limit: C. albicans DNA extracted from yeast (sensitivity 0.2 CFU/µL), spiked human blood (sensitivity 10 CFU/mL), and cloned fragment of ITS2 region (sensitivity 20 target copies/μL). The efficiency of ITS2 fragment-qPCR ranged from 89.67 to 97.07, and the linearity (R2) of the standard curve ranged from 0.992 to 0.999. The results showed that this ITS2-qPCR has a great potential as a molecular prototype model for the development of a test to be applied in clinical practice, greatly reducing the time of candidemia diagnosis, which is extremely important in this clinical setting.

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ABSTRACT In experimental infection with Strongyloides venezuelensis, the acute and recovery phases can be distinguished, unlike human infections caused by Strongyloides stercoralis. The objective of this study was to evaluate the production of anti-Strongyloides IgG antibodies and the recognition of immunogenic protein bands during the acute and the recovery phases in rats experimentally infected with S. venezuelensis. Rats were infected subcutaneously with 400 or 4,000 S. venezuelensis infective larvae. The acute phase was characterized by elimination of a large number of eggs in the faeces on days 6-14 post infection; the recovery phase was characterized by the resolution of the infection between days 30 and 35 post infection. Differences in IgG levels were observed in the acute and the recovery phases. Different antigenic fractions were recognized in both phases of infection. It is concluded that proteins within the 30-40 kDa range are immunoreactive markers for both the acute and the recovery phases in rats experimentally infected with S. venezuelensis, particularly using membrane antigen.

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ABSTRACT Paracoccidioides species cause paracoccidioidomycosis (PCM), a systemic mycosis highly prevalent in Brazil. Therapy of PCM has some issues that make studies for new therapeutic and vaccine targets relevant, such as the P. brasiliensis 60-kDa-heat-shock protein (PbHsp60), an immunogenic antigen that induces protection in experimental mice infection. Here, we investigated the relative expression of mRNA for PbHsp60 in P. brasiliensis in the different morphotypes of P. brasiliensis and in morphological transition phases. In addition, antibodies to rPbHsp60 were produced and used to analyze the location of PbHsp60 in yeast and hyphae by electron microscopy. The analyses showed a substantial increase in the relative amounts of HSP60 mRNA in yeast when compared to mycelium and an intermediate expression in transitional forms. Regarding the cell location, immunoelectron microscopy analysis revealed that PbHsp60 is within the cell wall. These observations suggest that this protein may be involved in the maintenance of the cell wall integrity and the interaction with the host for colonization, infection and pathogenesis.

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ABSTRACT Eleven lactating women were inadvertently vaccinated with 17DD yellow fever vaccine in a small city of Sao Paulo State, Brazil. Their infants were being exclusively breast-fed and the breastfeeding was interrupted for 10 days. Serum and breastmilk were collected from the vaccinated mothers and tested for the presence of genomic RNA of the vaccine strain 8, 10 and 15 days after vaccination. Viral RNA was not detected in any of the serum and human milk samples tested and the infants remained asymptomatic. Our result strengthens the effectineness of stopping breastfeeding for 10 days after the inadvertent yellow fever vaccination of lactating women.

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ABSTRACT Although the chemical control against leishmaniasis began in 1953 in Brazil, little information is available on how this strategy has affected populations of phlebotomine sandflies in the field. The objective of this study was to analyze the susceptibility profile of four populations of phlebotomine sandflies to the insecticide alpha-cypermethrin. Sandflies collected in field in four Brazilian municipalities were evaluated using CDC bottles in different concentrations of alpha-cypermethrin. A total of 1,186 phlebotomine sandflies were used in the bioassays. The LD50 ranged from 1.48 to 2.57 ug/mL in the field populations. For a dose of 5 ug/mL of alpha-cypermethrin, the LT50 and LT95 ranged from 17.9 to 27.5 minutes, and LT95 from 39.7 to 61.5 minutes, respectively. All the populations studied were highly susceptible to the insecticide alpha-cypermethrin. Routine studies are needed to detect changes in sandflies susceptibility to insecticides.

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ABSTRACT Age, sex and presence of comorbidities are risk factors associated with COVID-19. Hypertension, diabetes and heart disease are the most common comorbidities in patients with COVID-19. The objective of this study was to estimate the prevalence of patients with comorbidities who died of COVID-19 in Brazil. Searches of data were carried out on the official pages of the 26 State health departments and the federal district. The random-effect method was used to calculate the prevalence of patients with comorbidities who died. From the beginning of the pandemic in Brazil until May 20, 2020, 276,703 cases of COVID-19 were notified in Brazil, 6.4% died, 58.6% of whom were male. The prevalence of comorbidities among deaths was 83% (95% CI: 79 - 87), with heart disease and diabetes being the most prevalent. To our knowledge, this study represents the first large analysis of cases of patients with confirmed COVID-19 in Brazil. There is a high prevalence of comorbidities (83%) among patients who died from COVID-19 in Brazil, with heart disease being the most prevalent. This is important considering the possible secondary effects produced by drugs such as hydroxychloroquine.

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ABSTRACT Severe neurological problems and other special manifestations such as high prevalence of structural cardiac changes has been described in infants vertically exposed to the Zika virus (ZIKV) and has been called congenital Zika virus syndrome (CZS). Previous studies have shown that the 24-hour Holter heart rate variability (HRV) analysis allows the prediction of worse outcomes in infants with neurological impairment and higher risk of sudden infant death syndrome (SIDS), hypertension, diabetes mellitus and other cardiovascular diseases. This study describes the 24-hour Holter findings of infants with confirmed vertical exposure to the ZIKV by positive polymerase chain reaction (PCR) assays in the mother's blood during pregnancy and/or in the urine or cerebrospinal fluid of the newborn. Data analysis was descriptive and included two subgroups according to the presence of fetal distress, positive PCR to ZIKV in the newborn, CZS and severe microcephaly. Heart rate, pauses, arrhythmias, ST segment and QT interval analyses and HRV evaluation through R-R, SDNN, pNN50 and rMMSD were described. The Mann-Whitney test was performed to assess differences between the two subgroups. The sample consisted of 15 infants with a mean age of 16 months, nine of whom were male. No arrhythmias or QT interval changes were observed. The comparison of HRV through the Mann-Whitney test showed a significant difference between patients with and without CZS, with and without severe microcephaly, with lower HRV in the groups with severe microcephaly and CZS. The study suggests that there is an increased risk of SIDS and cardiovascular diseases in this group of patients.

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ABSTRACT Blastocystis sp. is a protist commonly found in stool samples of humans and animals. Biological and genetic factors of this organism remain controversial. The present study aimed to develop and implement the Blastocystis in vitro culture of Brazilian human isolates for routine use. The fecal isolates (n = 20) were maintained in our laboratory by several passages in Pavlova's medium. Cultures were monitored every 72 h by light microscopy. Genomic DNA was extracted to identify the subtypes (STs). In most isolates, the vacuolar form was prevalent. The amoeboid, granular and cystic forms were observed during in vitro cultivation. STs 1, 2, 3, 4 and 7 were identified. Our preliminary results show the generation time and forms present in the in vitro culture of Blastocystis subtypes isolated from Brazilian human isolates. Therefore, we emphasize the use of in vitro culture as a tool in future studies for the better understanding of the biological aspects of Blastocystis sp.

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ABSTRACT Human cytomegalovirus (HCMV) infections remain a neglected public health issue. The aim of the present study was to evaluate the frequency of HCMV congenital infections in newborns up to 1 month in the Sao Paulo State, from 2010 to 2018. The molecular characterization of HCMV-positive samples was also undertaken. Urine samples from 275 potential congenital HCMV-infected patients were tested by real-time Polymerase Chain Reaction (qPCR). HCMV-positive samples were amplified by conventional PCR targeting the UL89 gene, sequenced and searched for mutations. A total of 32 (11.6%) positive-HCMV cases were detected (mean Ct 30.59); mean and median age of 10.3 and 6 days old, respectively. Children aged between 0-3 weeks had higher HCMV detection rates (84.4%; 27/32). UL89 gene was successfully sequenced in two samples, both classified as the human betaherpesvirus 5. No described resistance-associated mutations were identified. A routine screening in newborns coupled with the genetic characterization of key viral genes is vital to decrease sequels associated with congenital HCMV infections.

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ABSTRACT Lacaziosis is a cutaneous mycosis caused by the fungus Lacazia loboi, described in different countries of Latin America and prevalent in the Amazon region. The ineffective immune response against the agent seems to be related to a Th2 pattern of cytokines. There are few reports exploring elements of the humoral response in these lesions. Our aim was to investigate some elements focusing on B cells, plasma cells and local expression of IgG and IgM antibodies. Forty skin biopsies of lower limbs were selected. The diagnosis of lacaziosis was based on direct mycological examination and histological analysis. The visualization of fungal cells was improved by using Gridley’s staining. An immunohistochemical protocol was performed to detect the expression of B cells, plasma cells, IgG and IgM. A double staining was performed to explore the presence of yeasts in the cytoplasm of keratinocytes, using an anti-AE1 AE3 antibody over Gridley’s staining. The inflammatory infiltrate consisted of macrophages, multinucleated giant cells, lymphocytes, and fibrosis. Fungal cells were frequent in the stratum corneum and in both, the dermis and, in 50% of the specimens, also in the epidermis. Cells expressing IgG were more abundant when compared to cells expressing IgM. B cells and the presence of IgG might indicate that the humoral response promotes a Th2 immune response resulting in an anti-inflammatory phenotype. Our results lead us to suggest a possible role of B cells and immunoglobulins in the mechanisms of lacaziosis pathogenesis.

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ABSTRACT Bartonella spp. are re-emerging and neglected bacterial pathogens. The natural reservoirs for several species of this genus are domestic animals such as cats and dogs, the most common pets in the USA and Brazil. Some cat studies suggest that the infection is more prevalent in tropical and poverty-stricken areas. These bacteria were associated with a wide spectrum of clinical manifestations: fever of unknown origin, endocarditis, angiomatosis, chronic lymphadenopathy, hepatitis, fatigue, paresthesia and pain. Our group has already demonstrated that B. henselae -infected sickle cell disease mice present with hyperalgesia. We hypothesized that even immunocompetent mice infected by B. henselae would show an increased and persistent mechanical sensitivity. Five ten-week old male BALB/c mice were intraperitoneally inoculated with a 30 µL of suspension containing 10 4 CFU/mL of B. henselae, while five others were inoculated with an equal volume of saline solution. Four days after bacterial inoculation, the mechanical paw withdrawal threshold was measured using von Frey filaments in all animals, for five consecutive days. The infected animals showed hypersensitivity to mechanical stimuli for five consecutive days. The present study has demonstrated that B. henselae infection induces persistent mechanical hypersensitivity, a signal consistent with pain.

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ABSTRACT Chloroquine is the first-line therapy against the asexual stages of Plasmodium vivax . There is a high variation of chloroquine plasma levels after therapeutic doses, which can lead to inadequate exposure to the drug. The gender influence was low regarding the disposition of the drug, which is relevant as there are significant physiological variations between male and female patients. The objective of the study was to investigate whether gender modifies the pharmacokinetics parameters of chloroquine in patients with malaria vivax. A prospective study was performed in male and female adult patients using chloroquine (total dose of 25 mg/kg for three days) combined with primaquine. Serial blood samples were collected at admission and up to 672 h post-administration of the drugs. Chloroquine was measured in plasma samples by high-performance liquid chromatography with fluorescence detection. A non-compartmental analysis was used for modeling the data. A total of 26 male and 25 female patients were enrolled in the study. The pharmacokinetic parameters of chloroquine were similar between male and female patients: a half-life of 9.5 days and 10.2 days, maximum concentration (Cmax) of 1295 ng/ml and 1220 ng/ml, area-under-the-curve (AUC 0–28) of 241 µg/mL h and 237 µg/mL h, observed clearance (CL/f) of 5.8 and 5.5 L/h and the volume of distribution (V/f) of 1869 L and 1936 L. The study results suggest that a similar dose regimen of chloroquine combined with primaquine provides a comparable pattern of exposure in male and female patients.

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ABSTRACT The etiological agent of American trypanosomiasis is the protozoan Trypanosoma cruzi , typically transmitted by triatomines. The aim of this study was to investigate the triatomine fauna and trypanosomiasis infections in Acre State , Western Brazilian Amazon. Insect collection was performed by dissecting palm trees and installing traps. We found that T. cruzi infection rate was 24.5% and Rhodnius pictipes (57.1%) was the most abundant triatomine species. Health education as well as epidemiological and entomological surveillance are necessary to diagnose and prevent new cases of Chagas disease in the region.

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ABSTRACT Campylobacter is not routinely tested in foodborne disease investigations in Brazil. Here, we studied the occurrence of Campylobacter among other food-related bacteria commonly found in foodborne disease outbreaks reported in Rio Grande do Sul State, Southern Brazil. Seventy-two food samples were analyzed by using culture-based detection methods during the 18-month investigation of 36 foodborne disease outbreaks. The sampled foods from the foodborne disease outbreaks were all negative for Campylobacter . However, at least one of other routinely investigated foodborne-related bacteria was detected in 29.17% (21/72) of the samples. Taken together, these results suggest the need to monitor Campylobacter in foodborne diseases to detect sporadic cases caused by Campylobacter that might go unnoticed in Rio Grande do Sul.

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ABSTRACT Abietane diterpenoids are naturally occurring plant metabolites with a broad spectrum of biological effects including antibacterial, antileishmanial, antitumor, antioxidant, as well as antiinflammatory activities. Recently, we found that some analogues of natural ferruginol ( 2 ) actively inhibited dengue virus 2 (DENV-2) replication. Due to the similarity with DENV, we envisaged that abietane diterpenoids would also be active against Zika virus (ZIKV). Six selected semi-synthetic abietane derivatives of (+)-dehydroabietylamine ( 3 ) were tested. Cytotoxicity was determined by MTT assay in Vero cells. In vitro anti-ZIKV (clinical isolate, IMT17) activity was evaluated by plaque assay. Interestingly, these molecules showed potential as anti-ZIKV agents, with EC50 values ranging from 0.67 to 18.57 µM, and cytotoxicity (CC50 values) from 2.56 to 35.09 µM. The 18-Oxoferruginol (8) (EC50 = 2.60 µM, SI = 13.51) and 12-nitro-N-benzoyldehydroabietylamine (9) (EC50= 0.67 µM, SI = 3.82) were the most active compounds, followed by 12-hydroxy-N-tosyldehydroabietylamine ( 7 ) (EC50 = 3.58 µM, SI = 3.20) and 12-hydroxy-N,N-phthaloyldehydroabietylamine ( 5 ) (EC50 = 7.76 µM, SI = 1.23). To the best of our knowledge, this is the first report on anti-Zika virus properties of abietanes.

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ABSTRACT Malaria is an infectious vector-borne disease with other important routes of transmission, such as blood transfusion and organ/tissue transplantation, due to asymptomatic reservoirs of Plasmodium presenting with low parasitemia. Reports of transfusion-transmitted malaria have shown that in immunosuppressed recipients, infections can be fatal if they are not diagnosed and timely treated. All Plasmodium species can survive on blood components at temperatures from 2 to 6 °C for some days or even weeks. This report describes two candidates for blood donation harboring Plasmodium, infected in an area considered non-endemic. Blood samples were collected from donors who attended a blood bank in Sao Paulo and tested by microscopy, qPCR for Plasmodium genus-specific amplification, targeting the parasite 18S ribosomal subunit gene and a multiplex qPCR based on mtDNA of the five species. Under microscopy, only structures resembling Plasmodium were observed. The qPCR whose standard curve tested parasites varying from 2 to 0.1 parasites/ µL, showed the presence of Plasmodium DNA in the two blood donors, as did the multiplex qPCR that revealed the presence of P. malariae. The prevalence of positive donors varies according to the level of transmission, ranging from 0.7 to 55% in endemic areas. In non-endemic regions, prevalences are lower, however, transfusion malaria can evolve to severe cases, due to the lack of suspicion of this transmission route. Asymptomatic donors from low transmission regions pose a risk to blood banks, with particular emphasis on those located in areas with malaria elimination goals.