Doutorado em Doenças Infecciosas

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HIV-AIDS e co-morbidades não infecciosas : alterações ósseas, renais metabólicas em pacientes infectados pelo HIV-1 atendidos em serviço de assistência especializada em Vitória, ES
(Universidade Federal do Espírito Santo, 2012-09-14) Pinto Neto, Lauro Ferreira da Silva; Rodrigues, Rodrigo Ribeiro; Miranda, Angélica Espinosa Barbosa; Hadad, Davi Jamil; Dietze, Reynaldo; Tupinambas, Unai; Greco, Dirceu Bartolomeu
The increase in life expectancy achieved following modern antiretroviral exposes the HIV-infected population to non-infectious co-morbidities that worsen with ageing. The aim of this work was to study prevalence for renal, bone and metabolic disturbances and associated factors among HIV infected patients in an AIDS outpatient clinic. Urine samples exams, creatinine, glucose and lipids measurements, assessment of cardiovascular risk, HIV viral load, TCD4/TCD8 cell count and were done in patients attended by specialized assistance service at Santa Casa de Vitoria. Bone mineral density by dual-energy x-ray absorptiometry was performed at CEDOES (Center for Diagnostic and Research on Osteoporosis of Espirito Santo). Four papers were written based in project’s results: a case report of spontaneous supracondylar femur fracture in a patient with well controlled HIV viral load; an original report of low bone mineral density among 54,7% out of 300 patients, associated with menopause, male gender and body mass index bellow 25; another report of 40,6% of urinary abnormalities and 9,8% of reduced glomerular filtration out of 254 patients, the later one associated with black race, arterial hypertension, age above 50 years, proteinuria and hematuria; and a report of diabetes, impaired glucose tolerance and cholesterol disturbances among 5,8%, 28,5% and 22,3%, respectively, out of 498 HIV treated patients, the later one associated with lopinavir/r therapy. We conclude that clinical management of those age related diseases is essential for appropriate treatment of HIVinfected patients.
Mortalidade por cirrose hepática e etiologia da cirrose e do carcinoma hepatocelular no Espírito Santo : participação dos vírus B e C das hepatites e do alcoolismo crônico
(Universidade Federal do Espírito Santo, 2013-03-01) Gonçalves, Patrícia Lofêgo; Pereira, Fausto Edmundo Lima; Carrilho, Flair José; Coelho, Henrique Sergio Moraes; Dietze, Reinaldo; Miranda, Angélica Espinosa Barbosa
abstract
Estudo molecular de vírus associados à diarreia em crianças quilombolas do norte do Estado do Espírito Santo
(Universidade Federal do Espírito Santo, 2013-08-29) Vicentini, Fernando; Leite, José Paulo Gagliardi; Spano, Liliana Cruz; Mascarenhas, Joana Darc Pereira; Palaci, Moises; Miranda, Angélica Espinosa Barbosa; Fumian, Túlio Machado
Diarrhea is a major public health problem and its consequences are aggravated by lack of medical care, diagnosis and treatment support, greatly exposing children to death. This study aimed to conduct a surveillance between 2007 and 2010 followed by detection and molecular phylogenetic analysis of viral agents found in the feces of children with (symptomatic) and without diarrhea (asymptomatic), under 12 years, and the susceptibility to norovirus infection according to blood group antigen (HBGA). A total of 574 fecal samples were obtained and searched for: (i) rotavirus (RV), by polyacrylamide gel electrophoresis and enzyme immunoassay (ii) norovirus (NoV), by RT-PCR of the B viral genome region and genotypes and genogroups determined by PCR amplification and sequencing, respectively, for the regions C and D; (iii) adenovirus (HAdV), by PCR and nested-PCR for the hexon gene, species and genotype analysed by restriction enzyme SalI and sequenced and; (iv) astrovirus (AstV), by real-time PCR for ORF2. HBGA phenotypes were determined by gel-spinning and FUT2 and FUT3 genes, analyzed by PCR and sequencing from blood and saliva, respectively, obtained from children infected with NoV and adults.
Estudo sobre características genéticas de Mycobacterium tuberculosis isolados de pacientes com e sem lesões cavitárias
(Universidade Federal do Espírito Santo, 2013-08-30) Vinhas, Solange Alves; Suffys, Philip Noel; Palaci, Moisés; Nunes, Ana Paula Ferreira; Spano, Liliana Cruz; Gomes, Harrison Magdinier; Kritski, Afrânio Lineu
Background: Based on the hypothesis that genetic variability of Mycobacterium tuberculosis (MTB) could influence virulence and immunopathology we analyzed genetic profiles of different MTB strains in order to detect relatedness between genetic diversity and presence of cavity (disease severity). Methods: We conducted a retrospective molecular study in Vitória ES, based on TB strains (2003 to 2006, n = 214) from patients with pulmonary cavitary and non-cavitary TB using IS6110-RFLP, Spoligotyping and MIRU-VNTR methodologies. RESULTS: Initially, we compared the association of the demographic and clinical characteristics of patients with the presence of cavities. After logistic regression the variables that most contributed to explain the model of the disease were smear positive (ORajust = 5.96; IC= 2.58-13.73) and sputum production (ORajust = 4.55; IC= 1.28-16.12), there was no statistically significant association with the remaining variables. The LAM family was the most frequent within the samples of the two groups analyzed, representing 65 (62%) of the isolates in the cavitary group and 40 isolates (38%) of the non-cavitary. After comparing the proportions of LAM and other spoligotyping families there was no statistically significant difference between the groups (p=0.17). In relation to deletions RDRio (p=0.65) and RD174 (p=0.65) there were no statistically significant difference between the groups. Amongst the 205 isolates analyzed, 25 (12%) belonging to the non-cavitary group and 43 (21%) belonging to the cavitary group, were grouped in clusters. The statistical analysis of the association of the occurence of clusters with the presence of cavity showed no statistically significant difference between the quantity of clusters and the groups that were analyzed, (p= 0.4). Conclusion: The genotipic profile for the isolates from patients with cavitary and non-cavitary disease was determined. Our data showed that LAM9 was the most frequent among the strains between cavitary and noncavitary groups, corroborating findings that this family is the most frequent in Brasil. There were no statistical differences that could show association among the variables analyzed related to presence of cavity or disease severity
Caracterização molecular de papilomavírus humano (HPV) e vírus adeno-associado (AAV) em lesões intraepiteliais de colo uterino : um estudo de seguimento
(Universidade Federal do Espírito Santo, 2014-06-04) Freitas, Luciana Bueno de; Miranda, Angélica Espinosa Barbosa; Spano, Liliana Cruz; Louro, Iuri Drumond; Musso, Carlos; Rodrigues, Rodrigo Ribeiro; Palaci, Moisés
The cervical cancer (CC) is one of the most frequent types of cancer in women worldwide, not only in incidence but also in mortality, with HPV as the etiological agent. Some HPV genotypes, termed high-risk (HR-HPV) and its gene variants are more associated with the induction of malignancy in the cervix, being HPV16 and 18 the most frequent types. Some infections of the genital tract may act as cofactor of carcinogenic progression of CC, but adeno-associated virus (AAV) infection seems to be inversely related, which may reflect a protective role in the development of HPV-induced cervical cancer. Therefore, this study aimed to investigate the role of AAV-HPV mixed infection and oncogenic variants of HPV in the progression of cervical intraepithelial and to follow the viral persistence / clearance and the relation to the progression / regression of cervical lesions. Cytological examinations were performed on cervical samples, collected in two moments, of women referred to the University Hospital Cassiano Antonio Moraes – HUCAM, and all of them received treatment as recommended. DNA was extracted using QIAamp® DNA Mini Kit commercial kit, following manufacturer's instructions. AAV DNA was investigated by PCR and nPCR and HPV by PCR and Hybrid Capture® (CH). AAV and HPV genotyping were performed by RFLP and RLB, respectively. From all the cases referred to the colposcopy clinic, 57.3% had normal cytology, 23.1% low grade intraepithelial lesions and 19.6% high-grade intraepithelial lesions. Out of the normal groups, 78% remained normal, while 22% progressed to lesion; from the cases with low-grade lesion, 74% regressed to normal cytology, while 78.6% of the cases with high-grade lesion showed low-grade lesion or normal cytology in the second collection. Fifty six percent and 36.5% of the samples were positive for HPV in the first and second samples, respectively. A good correlation (kappa = 0.66) between CH and PCR tests for HPV detection was observed. HR-HPV was detected in about 90% of the samples from both collections, the most common types were HPV16, 58, 51, 52 and 53. Non-European variants were associated with the development of high-grade cervical lesions, while AAV presence was inversely related to the progression of HPV-induced cervical lesions.
Análise de transmissão e da dinâmica de modificação de genótipos de Mycobacterium turbeculosis na Região Metropolitana de Vitória-ES em um intervalo de 10 anos
(Universidade Federal do Espírito Santo, 2015-02-25) Nóbrega, Renata Lyrio Peres; Palaci, Moisés; Maciel, Ethel Leonor Noia; Oliveira, Martha Maria de; Nunes, Ana Paula Ferreira; Miranda, Angélica Espinosa Barbosa; Suffys, Philip Noel
Introduction: Molecular epidemiology have contributed to the understanding of the dynamic of transmission of tuberculosis (TB). Although many papers have been published on this subject, few studies, however, were performed taking into account the dynamic of modification of genotypic profiles of M. tuberculosis (Mtb) in a long time frame. Objective: To identify genotypes and factors associated with cluster size of Mtb in the population of the Metropolitan Area of Vitória – ES (RMV) and to analyze the dynamic of modification of Mtb genotypes in the RMV in a 10 years frame. Methods: This study had two parts.
Análise espacial da transmissão de genotipos de Mycobacterium tuberculosis em Vitoria, ES-Brasil
(Universidade Federal do Espírito Santo, 2015-02-26) Ribeiro, Fabíola Karla Corrêa; Maciel, Ethel Leonor Nóia; Palaci, Moisés; Suffys, Philip Noel; Oliveira, Martha Maria de; Falqueto, Aloísio; Spano, Liliana Cruz
Background. Genotyping Mycobacterium tuberculosis isolates allows study of dynamics of tuberculosis (TB) transmission, while geoprocessing allows concomitant spatial analysis of clinical and epidemiological data. In the present study, genotyping data and spatial analysis were combined to characterize TB transmission in VitóriaES-Brazil to identify distinct neighborhoods and risk factors associated with recent TB transmission. Methods. From 2003 to 2007, 503 isolates were genotyped by IS6110 restriction fragment length polymorphism (RFLP) and spoligotyping. The spatial analysis included Kernel density estimation (KDE), k-function analysis on predicted estimates from a random-effects logit model and a t-test distance analysis. M. tuberculosis isolates belonging to identical RFLP patterns (clusters) were considered to represent recent TB infection (cases). Results. Of 503 genotyped isolates, 242 (48%) were categorized into 70 distinct clusters belonging to 12 RFLP families. The overall proportion of recent transmission was 34.2%. Kernel density maps indicated three areas of most intense concentration of cases. K-function analysis of the largest RFLP clusters and families showed that both co-localized in space. The distance analysis confirmed these results and also showed that unique-pattern strains (controls) randomly distributed in space. A logit model with random neighborhood effects was used to evaluate univariate and multivariate associations. When the predicted probabilities for each neighborhood were mapped, they identified the neighborhoods with high risk for recent transmission. Conclusions. Spatial and genotypic clustering of M. tuberculosis isolates revealed ongoing active transmission of TB caused by a small subset of strains in a subset of neighborhoods of the city. Such information provides an opportunity to target TB transmission control, such as through rigorous and more focused contact investigation programs.
Pesquisa de anticorpos anti-PGL-I em pacientes infectados pelo HIV em área endêmica para hanseníase
(Universidade Federal do Espírito Santo, 2015-02-26) Madureira, Brunela Pitanga Ramos; Deps, Patrícia Duarte; Falqueto, Aloisio; Palaci, Moises; Antunes, João Marcelo Azevedo de Paula; Hadad, David Jamil
The present study investigated subclinical Mycobacterium leprae infection in human immunodeficiency virus (HIV) infected or uninfected individuals by measuring antibodies against M. leprae phenolic glycolipid I (PGL-I). It also sought to establish whether the serology results correlated with the HIV-infected participants’ state of immunosuppression. This was a cross-sectional study that analysed the anti-PGL-I immunoglobulin M antibody levels in 350 HIV-infected and 350 non-HIV-infected individuals residing in an area endemic for the two investigated conditions. The possible correlation between the state of immunosuppression of the HIV-infected participants (CD4+ cell count, viral load and the use or not of antiretroviral therapy) and anti-PGL-I seropositivity was assessed. Approximately 6% (21/350) of the HIVinfected and 29.1% (102/350) of the non-HIV-infected participants were anti-PGL-I antibody seropositive. The number of anti-PGL-I antibody seropositive individuals was approximately five-fold higher in the non-HIV-infected group, compared with the HIV-infected group. There was no significant correlation between the state of immunosuppression of the HIV-infected participants and the anti-PGL-I serology results. The HIV-infected individuals exhibited less anti-PGL-I antibody production relative to the control group, which may indicate a lower rate of subclinical M. leprae infection and/or lower specific production of this antibody. B cell dysregulation might cause the low anti-PGL-I antibody production in HIV-infected individuals. The patients’ state of immunosuppression did not correlate with the anti-PGL-I serology results.
Prevalência de Chlamydia trachomatis em casos de partos pretermo atendidos no Hospital Universitário Cassiano Antonio Moraes, Vitória-ES
(Universidade Federal do Espírito Santo, 2015-03-24) Lopes, Renylena Schmidt; Silveira, Mariângela Freitas da; Miranda, Angélica Espinosa Barbosa; Paulo César Giraldo; Dietze, Reynaldo
Background: Premature birth (PPT) is a major determinant of neonatal morbimortality with adverse consequences for health. The causes are multifactorial, with intrauterine infection probably explains most of these outcomes. It is believed that infection with Chlamydia trachomatis (CT) is also involved in PPT and premature rupture of membranes (ROPREMA). Objetives: To study the prevalence of CT in pregnant women and possible risk factors related to cases of PPT attended at University Hospital Cassiano Antonio Moraes. Methods: A cross-sectional study performed among parturient who have preterm birth in an University Hospital in Vitória - ES, from June 2012 to August 2013. Participants answered a questionnaire including demographic, behavioral, and clinical data. A sample of urine was collected and screened for CT using polymerase chain reaction. Results: The prevalence of PPT in the hospital during the period of the study was 26%. A total of 378 cases of PPT were registered, among them 323 women participated and were tested for CT, forty-five (13.9%) had a positive result. 31.6% was up to 24 years old and women infected by CT were younger than the others (p=0.022). A total of 76.2% were married/living together, and CT was more frequent among the single ones (p=0.018); 16.7% of women had their first sexual activity under 14 years old. The causes of prematurity were maternal-fetal in 40.9%, rupture of the membranes in 29.7% and premature labor in 29.4%. In multivariate analysis, being married was a protective factor for infection [OR = 12:48 (95% CI: 0.24-0.97)]. None of the other characteristics were associated with CT infection. Conclusions: This study shows a high prevalence of CT infection among parturient who have preterm birth. This high prevalence increases the need for defining screening strategies and assistance during the prenatal period
Coorte retrospectiva de crianças e adolescentes, infectados pelo HIV por transmissão vertical, no Espírito Santo : comorbidades, mortalidade e sobrevida
(Universidade Federal do Espírito Santo, 2015-03-25) Silva, Sandra Fagundes Moreira da; Zandonade, Eliana; Miranda, Angélica Espinosa Barbosa; Pinto, Jorge Andrade; Machado, Daisy Maria; Dietze, Reynaldo; Cerutti Junior, Crispim
BACKGROUND: Early diagnosis and antiretroviral therapy in infants infected with HIV through vertical transmission, reduces HIV progression and comorbidities that can lead to death. OBJECTIVE: The goal of this study was to describe the frequency of comorbidities and to verify their distribution according to demographic, epidemiological and clinical data; to evaluate risk factors for progression to death, late diagnosis and trend of mortality in a cohort of children and adolescents infected by HIV vertical transmission in a reference hospital in the state of Espírito Santo. SPECIFIC OBJECTIVES: 1. To describe the frequency of comorbidity diagnosed after the diagnosis of HIV and verify their distribution according to demographic, epidemiological and clinical data, and according to the classification of cases in a cohort of children and adolescents with AIDS. 2. To evaluate the progression of predictors of risk factors for AIDS and death and cause of death. 3. To estimate the survival rate. METHODS: Retrospective cohort study of children and adolescents infected by HIV by vertical transmission (VT), treated at the Hospital Infantil Nossa Senhora da Glória (HINSG) from January 2001 to December 2011 in Vitória - ES / Brazil. Data collection was carried out in a specific protocol standardized, and data on comorbidities, mortality and its underlying causes were obtained from medical records, death certificates and SIM database (Mortality Information System). The aids diagnosis and comorbidities were according to CDC(Centers for Disease Control and Prevention)/1994. RESULTS: From a total of 177 patients, 97 (55%) were female and 60 (34%) were less than 1 year, 67 (38%) had 1-5 years and 50 (28%) were 6 or more years old at enrollment into the service. Median age at admission was30 months (IQR 25-75%: 5-72 months). In relation to clinical and immunological classification (146, 82.5%) had a moderate / severe status at the time of entry into service and 26 (14.7%) died. The most frequent clinical signs were hepatomegaly (81.62%), splenomegaly (63.8%), lymphadenopathy (68.4%) and persistent fever (32.8%). The most frequent comorbidities were anemia (67.2%), pneumonia / sepsis / Bacterial Meningitis (64.2%), Acute Otitis Media (AMO)/ recurrent sinusitis (55.4%), recurrent severe bacterial infections (47.4%) and dermatitis (43.1%). An association was found between clinical and immunological classification in a severe form and entry into service at less than one year old with few comorbidities (p<0.001). The total time of the follow-up of the patients was 11 years, median of five years (interquartils range, IQR: 2-8 years). At the end of the study period 132(74.6%) patients were followed up, 11 (6.2%) were transferred to other services and eight (4.5%) were lost to follow-up. Regarding death as an outcome we observed a reduction of cases over time. Most patients who died were admitted to the service with classification of severe immune clinical status (77 % -20/26) had moderate / severe anemia and were on HAART for more than 3 months (17/24-71 %). The main risk factors for death were: age <1 year (p=0.005), P.jirovecii pneumonia (p=0.010), the percentage of T lymphocyte CD4 + nadir <15% (p=0.012), chronic anemia (p=0.012), severe clinical and immune status(p=0.003), recurrent severe bacterial infections (p=0.003) and tuberculosis (p=0.037). Having started HAART before 6 months of life (early diagnosis and treatment) was associated with being alive (OR 2.86, [CI 95%: 1.12 to 7.25] p=0.027). The principal diagnosis recorded for deaths were severe bacterial infections (12/21-57%). A high survival rate was observed, with 85.3 % probability of survival for more than 10 years (95%CI: 9.6-10.7). CONCLUSIONS: Most children that had late diagnosis of HIV infection also had an increased the risk of progression to aids and death because of lack of early treatment. The trend of mortality of HIV-infected children showed a steady decline in the past two years of the study, and bacterial infections persisted as major cause of death. Therefore, improvements in prenatal care and pediatric follow-up with a view to early diagnosis of vertically infected children should be part of comprehensive care for children with AIDS which could reduce mortality of these children.
Estudo do mecanismo de resistência natural à miltefosina em isolados de Leishmania (Leishmania) chagasi obtidos de pacientes com leishmaniose visceral que apresentaram diferentes respostas ao tratamento
(Universidade Federal do Espírito Santo, 2015-06-16) Trindade, Juliana Brambilla Carnielli; Figueiredo, Suely Gomes de; Lemos, Elenice Moreira; Ruiz, Jeronimo C.; Borges, William de Castro; Pereira, Fausto Edmundo Lima; Spano, Liliana Cruz
Visceral leishmaniasis (VL) is a systemic disease that is fatal if untreated and is caused by the Leishmania donovani complex, which include the Leishmania (L.) chagasi. Visceral leishmaniasis treatment relies on a few chemotherapeutic drugs including Sb(V), amphotericin B and miltefosine. Miltefosine is the first oral drug registered for leishmaniasis treatment and it has been highly active against VL in India. However, susceptibility differences to miltefosine have been observed in clinically relevant Leishmania species. Miltefosine resistance mechanisms are being elucidated in laboratory Leishmania spp. isolates but are less clear in clinical isolates. In this study, we used a comparative proteomics and genomics approaches to highlight molecular differences between L. (L.) chagasi isolates from visceral leishmaniasis patients with different miltefosine treatment outcomes. The highresolution proteomes obtained from one isolate from a relapsed patient and the other isolate from a patient who relapsed after miltefosine treatment showed 46 spots that exhibited different abundances between the isolates. Out of these differentially expressed spots. MALDI/ToF-ToF mass spectrometry allowed the identification of 32 spots with unique protein identification that correspondent to 22 non-redundant proteins. Most of the proteins upregulated in the proteome of the isolate from relapsed patient were associated with redox homeostasis, stress response, protection to apoptosis, and drug translocation. The whole genome sequence carried out with isolates from patients who displayed cure (n=14) and relapse (n=12) clinical outcome, identified a high number of SNPs and InDels. However, same as the chromosome copy number variation analysis, no SNPs and InDels completely discriminated between analysed groups. Against a background of relative genetic homogeneity, we found significant variation (p < 0,01) in gene dosage between the isolates from cure and relapse groups: 93 orthologs groups (OG5). Within these, we assessed the association between the deletion of the in tandem genes LinJ.31.2370, LinJ.31.2380, LinJ.31.2390 e LinJ.31.2400 with the resistance phenotype of the L. (L.) chagasi. It was demonstrated that this deletion process occurs by homologous recombination, and apparently is not induced by miltefosine pressure. The individual reexpression of these genes did not interfere in the in vitro miltefosine susceptibility phenotype of promastigote stage. Furthermore, the clones separated from clinical isolates of L. (L.) chagasi (heterogeneous with respect to the presence of these genes) showed that the promastigote stage of clones that present these genes are less susceptibility to miltefosine than clones that are absent from these genes. These data suggest, same as the proteomics approach, that the natural miltefosine-resistance mechanism in Leishmania spp. is complex and multifactorial.
Modelagem de nicho ecológico das espécies de Phlebotominae e sua relação com a distribuição da leishmaniose tegumentar americana em região endêmica do Sudeste do Brasil
(Universidade Federal do Espírito Santo, 2016-11-29) Meneguzzi, Viviane Coutinho; Fux, Blima; Falqueto, Aloísio; Miranda, Angélica Espinosa Barbosa; Cerutti Junior, Crispim; Werneck, Guilherme Loureiro; Shaw, Jeffrey Jon
Cutaneous leishmaniasis (CL) is caused by a protozoan of the genus Leishmania and is transmitted by sand flies. The State of Espírito Santo (ES), an endemic area in the Southeast of Brazil, has shown considerably high prevalence in recent decades,allowing the expansion of the disease to unaffected areas. Computer tools, such as ecologic niche modelling (ENM), are useful for predicting potential disease risk. In this study, ENM was applied to species of sand flies and CL cases in ES to identify the principal vector and risk areas of the disease, aiming to understand the early origin and expansion of this disease. Sand flies were collected in 466 rural localities between 1997 and 2013 in the three hours after dusk using a combination of activeand passive capture. Insects were identified to the species level, and the localities were georeferenced. All medical records of autochthonous cases of CL attended at the University Hospital Cassiano Antonio de Moraes (HUCAM) between 1978 and 2013 were evaluated.
Alcoolismo e Strongyloides stercoralis : investigação de possíveis fatores associados à maior prevalência do nematoide em alcoolistas crônicos
(Universidade Federal do Espírito Santo, 2017-02-17) Ribeiro, Steveen Rios; Pereira, Fausto Edmundo Lima; Falqueto, Aloísio; Gomes, Daniel Claudio de Oliveira; Braga, Fabio Ribeiro; Leite, Gustavo Rocha
Introduction. Strongyloides stercoralis (Bavay, 1876) is an intestinal nematode that can proliferate in the host by autoinfection, and persist for decades without further exposure to exogenous infection. The prevalence of the parasite is poorly known in the state. Higher prevalence of S. stercoralis in chronic alcoholic patients than in non-alcoholics, attended at the same hospital, has been reported, but there are doubts about if there is a greater exposure to the parasite or greater survival and fecundity of females in the duodenum would increase the chance of encountering larvae in the feces. Objectives. To evaluate: (a) the prevalence of S. stercoralis in the state on samples of schoolchildren aged 7 to 14 years and samples from patients attended at the different outpatient clinics of the Hospital C.A. Moraes (HUCAM) and Hospital Concórdia at Sta. Maria de Jetibá; (b) the prevalence of the parasite on samples of alcoholics and non-alcoholics in the Department of Gastroenterology of HUCAM and Hospital Concordia; (c) the number of S. stercoralis rabditoid larvae on stools of alcoholics and non-alcoholics patients; (d) the anti-S. stercoralis antibodies on serum and Treg lymphocytes (Treg) in peripheral blood in alcoholics and non-alcoholics; (e) the use of the conventional PCR method on stool samples of alcoholic and non-alcoholic patients, to confirm the difference in prevalence observed between the two groups.
"Ocorrência de Ehrlichia spp., Anaplasma spp., Babesia spp., Hepatozoon spp. e Rickettsia spp. em cães domiciliados em seis municípios do Estado do Espírito Santo, Brasil"
(Universidade Federal do Espírito Santo, 2017-07-20) Vieira, Fernanda de Toledo; Dietze, Reynaldo; Braga, Fabio Ribeiro; Moraes Filho, Jonas; Santos, Kênia Valéria dos; Palaci, Moises
Tick-borne diseases are an important cause of morbidity and mortality in dogs (Canis lupus familiaris) worldwide, with the tick Rhipicephalus sanguineus sensu lato (s.l.) as a vector of various agents. It is very important to know the prevalence of these diseases, their vectors and hosts in order to control and prevent them in the given region. Tick-borne diseases have importance for both veterinary medicine and for public health. This work aims to identify and quantify the percentage of infection by Rickettsia spp., Ehrlichia spp., Babesia spp. and Hepatozoon spp. in pet dogs and identify ticks collected on these animals in the municipalities of Santa Teresa, Colatina, Alegre, Vila Velha, Vitória and Serra. The study included 378 dogs, 226 females and 152 males, of various breeds and ages (mean 4.1 years) with ectoparasites (ticks) or parasitism recent history (30 days). Of the 378 dogs examined, 157 (41.53%) had ticks at the time of consultation, which were identified as R. sanguineus s.l. in 154 animals (98.1%), Amblyomma ovale in one animal (0.63%) and Amblyomma sculptum in 2 animals (1.27%). All animals were examined and whole blood were collected and processed by molecular tests: Real Time PCR for Ehrlichia canis, and conventional PCR protocols for Babesia spp., Hepatozoon spp., and for Anaplasmataceae. In the Real Time PCR specific for E. canis, 28 animals (7.4%) were positive. For Anaplasmataceae, 34 animals (9%) were positive, of which 10 generate DNA sequences with 100% similarity with Ehrlichia canis and the other 24 samples generated fragments 100% identical to Anaplasma platys. In the PCR for Babesia spp., 5 animals (1.3%) were positive, producing DNA sequences 100% identical to Babesia vogeli. Finally, 39 animals (10.31%) were positive by PCR for Hepatozoon spp., producing sequences 100% identical to Hepatozoon canis. It was observed coinfection with 2 or 3 agents in 22 animals (5.8%). Of the 378 dogs in the study, 312 had serum samples collected for serological tests: indirect immunofluorescence assay (IFA) for E. canis, with positive reaction in 71 animals (22,75%) and five species of Rickettsia (R. rickettsii, R. parkeri, R. amblyommatis, R. rhipicephali and R. bellii). Among the sera analyzed by IFA, 16 animals (5.13%) had titers ≥ 64 for at least one species of Rickettsia of the five tested, and 5 (1.6%) of those had titers ≥ 64 for R. rickettsii. These results indicate that some of the studied municipalities have agents transmitted by ticks circulating in the canine population, such as E. canis, A. platys, B. vogeli, H. canis, and possibly spotted fever group rickettsiae.
Mortalidad y predictores de mortalidad en niños menores de 15 años que adquirieron el VIH por transmisión madre-hijo en el Paraguay
(Universidade Federal do Espírito Santo, 2017-11-30) Barreto, Gloria Celeste Aguilar; Munoz, Sergio; Miranda, Angélica Espinosa Barbosa; Cerutti Junior, Crispim; Combadão, Jaime Manuel Pinto; Silva, Sandra Fagundes Moreira da; Saraceni, Valéria
Introduction: Efforts have been made in Paraguay to improve the response to HIV infection. In the context of the Sustainable Development Goals, it is important to evaluate the impact of interventions in pediatrics. Objective: To estimate the mortality rate and predictors in children who acquired HIV through mother-to-child transmission between January 2000 and December 2014. Material and methods: An observational, cohort-type study was performed with follow-up from the birth of children and adolescents who were <15 years of age at the time of recruitment. Data were obtained from clinical records. Death data were recorded from medical records and confirmations of deaths from death certificates. Survival analysis techniques were used to estimate the incidence of death Results: A total of 302 subjects were included, 216 (71.6%)% under six years of age,148 ( 51.0%) male, and 214( 70.9%)% of the main metropolitan area. There were 52 deaths (17.2%) with an overall mortality rate of 2.06 deaths / 100 personyears. Children with hemoglobin ≤9 g/dL were 2 times more likely to die compared to hemoglobin > 9 g / dL [ (hazard ratio; 2.27, (95% CI: 1.01, 5.10) ]. The risk of death was 74% lower for children diagnosed in the period ≥ 2010 compared to those diagnosed before 2010 [hazard ratio ; 0.26, (95 CI%, 0.07-0-88)] . Conclusion: The mortality of children with HIV in Paraguay is high, and anemia is associated with mortality. Improving prenatal care and pediatric follow-up in an effort to early diagnose children infected by mother-to-child transmission should be an integral part of the health care offered to the child with HIV infection.
Avaliação dos portadores assintomáticos de DNA de Plasmodium SP em área endêmica de malária no Estado do Espírito Santo
(Universidade Federal do Espírito Santo, 2017-12-11) Alencar, Filomena Euridice Carvalho de; Miranda, Angélica Espinosa Barbosa; Maciel, Ethel Leonor Noia; Alves, Rosana; Falqueto, Aloisio; Deps, Patrícia Duarte
abstract
Classificação morfológica e genotípica e correlação entre propriedades fisiológicas e citopatogênicas de isolados clínicos e ambientais de Acanthamoeba
(Universidade Federal do Espírito Santo, 2018-03-21) Possamai, Cynara Oliveira; Bueloni, Cinthia Furst Leroy Gomes; Falqueto, Aloísio; Fux, Blima; Gonçalves, Sarah Santos; Leite, Gustavo Rocha; Dettogni, Raquel Spinassé
Free living amoeba of the genus Acanthamoeba can eventually act as parasites, causing severe infections in humans and other animals. Some biological and physiological characteristics have been related to the grade of pathogenicity of strains, allowing inferences about the pathogenic potential of the genera. The main goal of this study was to classify isolates of Acanthamoeba obtained in Brazil, evaluate properties associated with their pathogenicity and discuss the results in association with the origin of the isolates. A total of 39 Acanthamoeba isolates obtained from amoebic keratitis cases (n=16) and environmental sources (n=23) were classified into morphological groups (I, II, III) and genotyped (T1-T20) by sequencing the 18S rDNA fragments ASA.S1 and GTSA.B1. Samples were also tested regarding their thermotolerance, osmotolerance and cytopathogenicity in MDCK cells. Comparisons of clinical and environment isolates were performed by x2 test (p<0,05). Isolates were classified as follows: group I (T17, T18); group II (T1, T3, T4, T11); and group III (T5, T15), with the predominance of genotype T4 (22/39). Clinical isolates were genotyped as T3 (1/16), T4 (14/16) and T5 (1/16). The majority of isolates (38/39) were able to grow at 37 °C, but tolerance to 40 °C was more frequent among environmental samples. The tolerance to 1 M mannitol was infrequent (4/39), with three of these corresponding to clinical samples. There was no significant difference between the numbers of clinical and environmental isolates that presented tolerance to 40 C (p=0,06), to 1 M mannitol (p=0,14) and ability to cause cytopathic effects (p=0,59). Variable correspondence of these properties was observed among distinct genotypes. This study identified, for the first time, the genotypes T1, T15 and T18 in Brazil. It also indicated a weak association between the clinical origin of the isolates and physiological tolerance tests and cytopathogenicity, demonstrating that some in vitro parameters do not necessarily reflect a higher propensity of Acanthamoeba to cause disease.
Transmissão da malária residual de sistemas de Mata Atlântica no Espírito Santo : comportamento vetorial e caracterização molecular das espécies de Plasmodium circulantes
(Universidade Federal do Espírito Santo, 2018-03-22) Buery, Julyana Cerqueira; Fux, Blima; Cerutti Junior, Crispim; Leite, Gustavo Rocha; Curado, Izilda; Spano, Liliana Cruz; Palaci, Moises
In the Southern and Southeastern regions of Brazil, autochtonous malaria cases can be found close to the Atlantic Forest fragments. In Espírito Santo state, this disease is particularly frequent, and Plasmodium vivax is the parasite commonly recognized as the etiological agent of human infections. However, when spatial and temporal distances between the reported cases and the behavior of local insect vectors, especially the Anopheles (Kerteszia) cruzii, are observed the transmission cycle does not correspond to the traditional cycle of malaria. Thus, the hypothesis of the existence of a zoonosis, with infected simians maintaining the transmission, was established. Considering that there are obscure aspects regarding bromeliad-malaria, as it is called, the evaluation of the components of the transmission cycle is necessary. In the present study, the transmission of bromeliad-malaria in the rural endemic region of Espírito Santo is investigated based on two main pillars: behavioral analysis of the Anopheles vectors, mainly those of the subgenus Kerteszia, and comparison of the complete mitochondrial genome from several isolates of Plasmodium spp. The isolates were infecting humans, simians of the genus Allouata and mosquitoes Anopheles spp. from the same region. Light CO2 traps (CDC-CO2) were installed in open areas, inside the forest and at its edge, and Shannon traps were installed on the edge of the forest. The mitochondrial genome of Plasmodium spp. found in the different hosts was completely sequenced and compared in a haplotype network that included all sequences of P. vivax/simian mitochondrial genomes from samples from humans and simians from other regions of Brazil. A total of 1,414 anophelines were captured from 13 species. Anopheles (Kerteszia) cruzii DyarΛ continued to be the more captured species in the forest canopy and was also the vector with the highest prevalence of Plasmodium vivax infection according to molecular PCR techniques. The haplotype network indicated that humans and simians of the Atlantic Forest were infected by the same haplotype, but some isolates of humans are not identical to the simian isolate. In addition, DNA of Plasmodium spp. extracted from mosquitoes presented sequences different from those obtained from simians, but similar to those retrieved from two isolates from humans. This study revealed that the acrodendrophilic behavior of vectors, especially those infected, reinforces the hypothesis that the presence of P. vivax in these specimens arises from the blood of animals that live in the top of the trees, such as monkeys. The comparison of the mitochondrial genomes of the parasites shows that, in the Atlantic Forest, especially in Espírito Santo, parasites with similar molecular structures are shared by humans and simians. The recognized identity between P. vivax and P. simium, the shared haplotypes, and the participation of the same vector in the transmission of the infection to both host species indicates interspecies transfer of the parasites.
Células "natural killers" : mecanismo de regulação exercido pelo receptor KLRG1 e perfil de diferenciação e funcionalidade na Leishmaniose cutânea
(Universidade Federal do Espírito Santo, 2018-05-28) Covre, Luciana Polaco; Gomes, Daniel Claudio de Oliveira; Rodrigues, Rodrigo Ribeiro; Salgado, Breno Souza; Falqueto, Aloísio; Pereira, Fausto Edmundo Lima; Guimarães, Marco Cesar Cunegundes
abstract
A influência da modulação por TLR2, TLR4 e TLR9 na resposta de células T reguladoras em cultura de sangue periférico de indivíduos com infecção latente pelo Mycobacterium tuberculosis, desafiadas in vitro com o bacilo de Koch
(Universidade Federal do Espírito Santo, 2018-07-10) Silva, Flávia Dias Coelho da; Rodrigues, Ricardo Ribeiro; Zeidler, Sandra Lúcia Ventorin Von; França, Johara Boldrini; Palaci, Moises; Pereira, Fausto Edmundo Lima
Latent tuberculosis infection (LTBI) affects approximately a quarter of the world's population. During LTBI, M. tuberculosis (Mtb) survives in a state of dormancy, which reactivates latent infection, which resumes normal growth and metabolism. Macrophages / monocytes (MO) play a central role in the mycobacterial pathogenesis, since they are the main cellular niche for Mtb during infections. The protective immune response, which the MO are part of is influenced by suppressive mechanisms, among them the increase of the activity of T regulatory cells (Tregs). Tregs have the ability to control tissue damage by decreasing adequate control of mycobacterial replication, and may also be involved in the reactivation and dissemination of Mtb. Toll-like receptors (TLRs) participate in the response to the infection by detecting and regulating it, and TLR2, TLR4 and TLR9 are known to recognize components of Mtb, which influence the response to kinetics and cytokine production by infection. We sought to assess the influence of TLR2, TLR4 and TLR9 agonists and antagonist in peripheral blood and Mtbchallenged whole blood cultures of individuals with LTBI (TST+ group) relative to the negative control (TST- group), investigating the frequency of Tregs and MO cells, the microbicidal activity and the dosage of cytokine IL10, IL17, TGFβ and IFNγ among these groups. Higher frequency of MO (CD14+ CD16+ HLA-DR+ , CD14+ TLR2+ HLA-DR+ , CD14+ TLR4+ HLA-DR+ , CD14+ TLR9+ HLA-DR+ ) was observed in the peripheral blood of LTBI/TST+ individuals. In the action of TLR2, TLR4 and TLR9 agonists or of TLR9 antagonist, under the frequency of Tregs cells from Mtb-challenged whole blood cultures, there was a higher frequency of these cells in the TST+ group, which was reduced after the use of TLR9 antagonist (chloroquine). As regards the influence of Mtb infection on the cultures, the microbicidal activity was lower in the TST+ group. In cultures infected with Mtb and TLRs-modulated, there was a reduction of the microbicidal activity in the TST+ group, during stimulation with TLR2 agonist, and, in the same individuals, in the stimulus with TLR9 antagonist, it was observed the restoration of the microbicidal activity. As for the dosage of cytokine in the same cultures, there was a higher production of IL10, IL17 and IFNγ in the TST+ group, especially after modulation with chloroquine, compared to the TST- group. In summary, LTBI differs from the control TST- by the higher frequency of Tregs and MO and the lower microbicidal activity, whereas the TLR9 blockade, by the use of chloroquine, resulted in the reduction of Treg cell frequency, in the higher production of IL17, IFNγ and IL10 and in the improvement of the microbicidal activity of LTBI in relation to TST-.

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