Navegando por Orientadores "MASCARENHAS, Joana D'Arc Pereira"

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Análise filogenética de genes de provável origem não humana de rotavírus do grupo A em espécimes fecais de crianças com gastrenterite aguda provenientes de Belém, Brasil
(Universidade Federal do Pará, 2012) MAESTRI, Régis Piloni; MASCARENHAS, Joana D'Arc Pereira; http://lattes.cnpq.br/5156164089432435
Rotaviruses (RVs) are the main cause of acute viral gastroenteritis in both humans and young animals of species such as calves, horses, pigs, dogs, cats, and birds. The genetic diversity of RVs is related to a variety of evolutionary mechanisms, including point mutation, genome reassortment, and reassortment. The objective of this study was realized the molecular charaxterization of the genes that encode structural and nonstructural proteins in unusual RV strains. The clinical specimens selected for this study were obtained from children and newborns with RV gastroenteritis, who participated in research projects on viral gastroenteritis conducted at the Evandro Chagas Institute. Structural (VP1-VP4, VP6, and VP7) and nonstructural (NSP1-NSP6) genes were amplified from stool samples by the polymerase chain reaction and subsequently sequenced. Eight unusual RV strains isolated from children and newborns with gastroenteritis were studied. Reassortments between genes of animal origin were observed in 5/8 (62.5%) strains analyzed. These results demonstrate that, although rare, interspecies (animal-human) transmission of RVs occurs in nature, as observed in the present study in strains NB150, HSP034, HSP180, HST327, and RV10109. This study is the first of its kind conducted in the Amazon region and supports previous data showing a close relationship between genes of human and animal origin, representing a challenge to the large-scale introduction of RV vaccines in national immunization programs.
Acesso aberto (Open Access)
Análise molecular de rotavírus tipo G9 de crianças na Região Norte do Brasil
(Universidade Federal do Pará, 2016-10-31) GUERRA, Sylvia de Fátima dos Santos; SOARES, Luana da Silva; http://lattes.cnpq.br/0556695301015859; MASCARENHAS, Joana D'Arc Pereira; http://lattes.cnpq.br/5156164089432435
Group A rotavirus (RVA) is the most viral agent associated with acute gastroenteritis, responsible for about 200,000 deaths among children aged under five years annually. RVA belongs to Reoviridae family, Rotavirus genus, its genome is composed by double-stranded RNA (dsRNA) with 11 segments encoding 12 proteins, six structural (VPs) and six non-structural (NSPs). Each protein designating a specific RVA genotype, being VP7 protein responsible for G genotype and currently there are 32 genetic variants. G9 genotype emerged on a global scale in the 90s, a period before RVA vaccine introduction in Brazil that occurred in 2006, and is continuously detected until present day. This study aimed to describe the frequency and genetic constellation associated with the current G9 genotype in Northern Brazil. It was selected 50 samples collected between 1999 and 2013, being 45 G9P[8], 2 G9P[4] and 3G9P[6], for fecal suspension preparation and dsRNA extraction for further genome amplification and sequencing of nucleotides. It was observed that during pre-RVA vaccine introduction period G9 frequency rate was 43%, while after RVA vaccine introduction the most frequece obtained was 12.5% (2008 to 2010). Phylogenetic analysis of VP7 gene showed that all strains belong to lineage III of G9, observing aminoacidic substitutions in antigenic sites when compared with vaccine strains. It was demonstrated in VP4 gene that P[8] strains gathered in lineage III, whereas P[4] grouped into lineage V and P[6] strains into lineage I. All G9P[6] and G9P[4] samples were associated with DS-1 constellation, genogroup 2, while G9P[8] samples showed Wa constellation, genogroup 1, except for one sample showing NSP3 gene with DS-1 profile. G9 samples from Northern region analyzed were associated with the expected constellations described in other parts of the world, except for one G9P[8] sample that showed a genetic restructuration in NSP3 protein. In the present study the same G9 lineages have circulated during pre and post RVA vaccine introduction periods, and it was described aminoacidic substitutions in relevant antigenic regions, such as it was reported genetic restructuration phenomenon in one sample of this genotype, emphasizing the continuous monitoring of current genetic variants of RVA.
Acesso aberto (Open Access)
Avaliação epidemiológica, clínica e molecular de enteropatógenos causadores de diarreia aguda em crianças e adultos residentes na comunidade Quilombola do Abacatal, Ananindeua, Pará
(Universidade Federal do Pará, 2015) KAIANO, Jane Haruko Lima; MASCARENHAS, Joana D'Arc Pereira; http://lattes.cnpq.br/5156164089432435
Acute diarrheal disease is a major cause of morbidity and mortality in developing countries and one of the factors that contributes to the worsening of the nutritional status of children. This study aimed to evaluate the clinical, epidemiological and molecular profile of infections by viral and parasitic agents in children aged 0-10 years and those over 10 years of quilombo of Abacatal in the 2008-2010 period. Fecal samples from 294 children were collected in the age group 0-10 years and 81 individuals over 10 years, residents of the community Abacatal, Ananindeua, Pará, which had acute diarrhea board or without diarrhea (controls). The viral diagnosis was made by immunochromatographic and molecular tests and parasitological by Faust and Hoffman method. A total of 375 fecal samples were obtained from 177 individuals. The frequency of viral agents in this study were rotavirus group A rotavirus C and picobirnavirus group by 6.4% (24/375), 0.3% (1/375) and 1.3% (5/375 ), respectively.The polyacrylamide gel electrophoresis confirmed the presence of rotavirus in 23 of 10 samples (43.48%) having short profile of 13 (56.5%) long profile. The presence of intestinal parasites was observed in 272 (77.94%) samples, and the most common were Ascaris lumbricoides detected in 13.18% (46/349) of the samples, followed by Trichuris trichiura with 10.88% (38 / 349), hookworms with 4.01% (14/349) and Strongyloides stercoralis 1.72% (6/349). Of the 24 samples positive for rotavirus group A the following genotypes were detected: G2P [4] (12.50%, 3/24); G1P [8] (25.00%, 6/24), G3P [9] (29.20%, 7/24) and G12P [6] (33.33%, 8/24). Two new genotypes were detected for VP6 genes (I18) and NSP1 (A19) of rotavirus A. Nutritional assessment of 38 children was conducted, showing that 18 4% (7/38) presented malnourished. This study highlights the need to implement preventive actions in the community, including education measures for health, vaccination against rotavirus, and even the implementation of programs to control parasitic infestations.
Acesso aberto (Open Access)
Avaliação temporal e genética do rotavírus genótipo G2 circulante na Região Norte do Brasil antes e após a introdução da vacina contra rotavírus
(Universidade Federal do Pará, 2016-10-31) OLIVEIRA, Alessilva do Socorro Lima de; MASCARENHAS, Joana D'Arc Pereira; http://lattes.cnpq.br/5156164089432435
Rotavirus group A (RVA) is the most important cause of diarrhea, accounting for about 40% of morbidity and mortality related to this disease in children around the world before the introduction of the vaccine. After the introduction of the vaccine against the RVA in Brazil in 2006 genotype G2RVA he rose again, being detected in up to 82% of children under five years of age performed post vaccination studies, leading to questions about the protection afforded by the vaccine facing the G2 type, as well as the occurrence of a selective pressure vaccine. Little is known about the evolution and diversity of G2 genotype and the possible influence of the vaccine on this. To provide a better understanding of the flow and genetic diversity of RVA genotype G2, we perform the time of circulation analysis of genotype over 31 years and analysis of structural and non-structural genes from samples that have circulated over 20 years in northern region of Brazil. The temporal assessment of movement of different genotype circulating in this region has observed that the G2 type RVA presented over the years a cyclical pattern of occurrence that did emerge in a post deployment of the vaccine scenario, suggesting a natural fluctuation due to variations natural occurring over time. Phylogenetic analyzes showed that for VP7 lines G2 there is a continuous, responsible for a movement of rotation in the lines being detected two lines and three sublineages over 20 years. Three important substitutions in antigenic regions of VP7 (A87T, D96N and S213D) were identified in samples that circulated from the 90. These changes may have increased the capacity of the circulating strains in environments where there is vaccine coverage for RVA. All G2P[4] strains analyzed revealed a DS-1-like genome constellation: I2-R2-C2-M2-A2-N2-T2-E2-H2. However, several viral variants circulated during the study period. No differences were observed in the antigenic sites of the VP8 * and VP7 proteins between samples that circulated in the period before and after the introduction of the vaccine. For VP2 and VP3 genes was evident in some samples a strong correlation with animal genes. This study provides evidence of genetic diversity in G2 genotype RVA, suggesting that this type has natural characteristics fluctuation and its emergence after the implementation period of the vaccine is more directly associated with ecological characteristics of the virus than a vaccine pressure.
Acesso aberto (Open Access)
Caracterização molecular dos genótipos G e P de rotavírus tipo G9 provenientes de crianças com gastrenterite aguda na região metropolitana de Belém, Pará, no período de 1999 a 2007
(Universidade Federal do Pará, 2010-07-05) GUERRA, Sylvia de Fátima dos Santos; SOARES, Luana da Silva; http://lattes.cnpq.br/0556695301015859; MASCARENHAS, Joana D'Arc Pereira; http://lattes.cnpq.br/5156164089432435
Rotavírus (RV) is the principal viral agent associated with gastroenterites, being responsible for 39% of the diarrhea cases that need hospitalization and with 520.000 deaths among children under five years. It belongs the to Reoviridae family, genus Rotavirus, the genome consists of 11 segments of double-stranded RNA that encoding 12 proteins, six structural (VPs) and six non-structural (NSPs). The VP4 protein, together VP7 protein, compose the outer layer of the RV, defining genotypes P and G, respectively. Until now, at least 23 different G genotypes and 31 P genotypes have been established. The G9 genotype emerged in world scale, and is possibly associated with severe clinical manifestation, being generally associated with P[8] genotype. The G9 genotype has 6 distinct lineages and P[8] 4 lineages. The aimed of this study was the characterization of VP4 and VP7 genes of the RV G9 genotype, that circulated in the metropolitan region of Belém, in the period of 1999 to 2007. The viral dsRNA of 38 selected samples were extracted from fecal suspensions and submitted to electrophorese in gel of poliacrylamide to determine the electropherotypes, followed sequencing reaction. In the present investigation, was possible analyzed 32 selected samples, all of them G9P[8] genotype, with long electropherotype. Phylogenetic analysis of VP7 gene showed that all G9 strains belong to lineage 3 with high similarity indexes, with 8 nucleotide changes. However, only three amino acids changes were observed in the positions 43 (I→V), 66 (A→V) e 73 (Q→R), being the substitutions at positions 43 and 73 exclusive of the samples isolated in 2007 year. Phylogenetic analysis of VP4 gene revealed that all strains P[8] belong to lineage 3, with 15 nucleotide changes, that showed four amino acids substitutions at positions 108 (V→I), 172 (R→K), 173 (I→V) e 275 (K→R). The amino acids substitutions at positions 172 and 275 were exclusive of the strains isolated from 1999 to 2002. The samples of the present study showed high homology throughout the studied period. The strains isolated in 2007 were the most divergence either to VP4 as to VP7 gene. It is important the continuous surveillance of G9 genotype in the metropolitan region of Belém, to detected possible new genetic variants that can represent a challenge to the current strategies of immunization.
Acesso aberto (Open Access)
Detecção, epidemiologia e análise molecular de rotavírus, picobirnavírus e reovírus em aves de corte criadas em granjas na mesorregião metropolitana de Belém, Pará, Brasil
(Universidade Federal do Pará, 2012-08-29) SILVA, René Ribeiro da; SILVA, Artur Luiz da Costa da; http://lattes.cnpq.br/7642043789034070; MASCARENHAS, Joana D'Arc Pereira; http://lattes.cnpq.br/5156164089432435
This study aimed to investigate the occurrence of avian rotavirus (AvRV), picobirnavirus (PBV) and avian reovirus (ARV) in chickens raised on farms located in the Metropolitan mesoregion of Belém, Pará state, in the period of 2008 to 2011. For this purpose, 85 pools of fecal samples were collected from 37 farms belonging to eight counties. Viral RNA was extracted from fecal suspensions and subjected to polyacrylamide gel electrophoresis (PAGE) followed by RT-PCR. At least one sample was selected of each municipality wich positive result for nucleotide sequencing of the genes NSP4 (AvRV), RdRp (PBV) and S2 (ARV), and in the case of the PBV samples were cloned before sequencing. PAGE showed positive in 0/85 (0%) samples for AvRV group A, 13/85 (15.3%) samples for PBV and 01/85 (1.2%) samples for ARV. In the case of RT-PCR positive results was observed in 35/85 (41.2%), 42/85 (49.4%) and 28/85 (32.9%) of the samples for AvRV, ARV and PBV, respectively. Of the eight counties studied, seven showed positivity to PBV, and six for AvRV and ARV. Of the 37 farms studied the presence of these viral infections was observed in 19 (51.4%) to AvRV and ARV and 21 (56.8%) to PBV. NSP4 gene sequences had a similarity between 86.3 and 90.5% at the nucleotide level (nt) with prototypes from chicken and 93.5 and 100% similarity at the nt level when compared among them. RdRp sequences showed a high genetic heterogeneity with gene variants resulting between 56.1 and 100% similarity at the nt level with prototypes belonging to different species and sources of contamination and between 50.3 and 100% similarity at nt level when compared among them. S2 gene sequences analysis showed between 90.9 and 94.4% similarity at the nt level with chicken prototypes and 90.1 and 100% similarity at the nt level when compared each other. AvRV, ARV and PBV were detected in broiler farms of the Metropolitan mesoregion of Belém, being the detection by RT-PCR more efficient to detect at least one type of virus in the eight counties surveyed. Except for the PBV, which showed heterogeneous relationship with the prototypes used, the AvRV and ARV of this study related specifically to the samples obtained in birds. This is the first study involving the genomic sequencing of the AvRV, ARV and PBV in broilers in northern Brazil.
Acesso aberto (Open Access)
Ocorrência de rotavírus do grupo D em aves criadas em granjas na mesorregião metropolitana de Belém, Pará
(Universidade Federal do Pará, 2012-02-29) BEZERRA, Delana Andreza Melo; MASCARENHAS, Joana D'Arc Pereira; http://lattes.cnpq.br/5156164089432435
Rotaviruses infect human beings and various species of animals. They are composed of non-enveloped icosahedral particles formed by a genome of 11 segments of dsRNA. They are classified into seven groups designated from A-G. The rotaviruses of group D (RVs-D) have been documented in birds; however there are few studies available especially the RT-PCR detection data with obtainment of nucleotide sequence. The aim of this study is to investigate the occurrence of RVs-D among the birds raised in farms situated in the metropolitan mesoregion of Belem, Para-Brazil during the period between 2008 and 2011. 85 pools of faecal samples were collected from 37 farms in eight counties of the metropolitan mesoregion of Belem. The viral dsRNA were extracted from faecal suspensions and submitted to polyacrylamide gel electrophoresis (PAGE) followed by RT-PCR conducted using the specific primers for VP6 and VP7 genes of the RVs-D, one positive sample was collected from each county for nucleotide sequencing of VP6 and VP7 genes, with five samples being cloned. The PAGE showed positivity in 14/85 (16.5%) of the samples and the RT-PCR in 30/85(35%) of the samples. Among the eight counties studied, seven presented samples positive for RVs-D and the viral infection was observed in 17(45.9%) of the 37 farms in these counties. The age interval where the RVs-D was detected at higher rate was among birds aged between 16 and 30 days (23/37-62%) of the total at this age. the nucleotide sequences of the seven samples were classified as RVs-D with bootstrap of 100 corroborating with this grouping. the sequences of VP6 gene presented 90. 8-91.3% of similarity with the prototypes of RVs-D (05V0049) and when compared among themselves. The VP7 genes presented 87-96.1% similarity with the prototype of this group (94.1-100% similarity) when compared among themselves. This analysis is a pioneer of its kind in Brazil and in the world, and it seeks to expand the knowledge about RVs-D.
Acesso aberto (Open Access)
Pesquisa de rotavírus e endoparasitos em animais na comunidade quilombola do Abacatal, munícipio de Ananindeua, Pará
(Universidade Federal do Pará, 2010-05-31) MATOS, Jane Cecília Silveira de; MASCARENHAS, Joana D'Arc Pereira; http://lattes.cnpq.br/5156164089432435
Rotaviruses (RVs) are the leading cause of acute diarrhea in children of low age, but also in young animals of various species. Are excreted in faeces and transmitted the fecal-oral route. Studies show that it is very important for epidemiological investigations to characterize samples of RVs isolated from animals. Intestinal parasites also pose a serious public health problem, being a major factor of morbidity and mortality in children and protein-energy malnutrition arising frameworks of chronic diarrhea. This study aimed to identify and endoparasites RVs circulating in dogs, cats and chickens Community Quilombola of Abacatal, Ananindeua, Pará. For the years 2008 and 2009 were collected 202 fecal samples, from dogs (96/2002, 47.5%), cats (8/2002, 4%) and chickens (98/2002, 48.5%). All samples were subjected to electrophoresis on immunochromatography and polyacrylamide gel for the identification of RVs, but in both cases we obtained 100% negativity. For the identification of endoparasites samples were subjected to a flotation technique with sucrose solution, the parasites most frequently found in dogs were Ancylostoma sp Spirocerca sp, Toxocara sp / Toxascaris sp, Trichuris sp and coccidio, and cats were Ancylostoma sp (5/8, 62,5%), Toxocara sp / sp Toxascaris (2/8, 25%) and Trichuris sp (1/8, 12,5%). In chickens were Ascaridia sp / sp Heterakis (33/62, 53,23%), Capillaria sp (39/62, 62,9%), Coccidio (6/62, 9,68%), Dispharynx sp (15/62, 24,19%) and Trichostrongyloidea sp (11/62, 17,74%). From the results, we concluded that the site poses a risk to human parasitic infestation, with the possibility of soil contamination by faeces and the development of zoonoses.
Acesso aberto (Open Access)
Rotavírus do grupo C associado à hospitalização de crianças com gastrenterite aguda em Belém, Pará
(Universidade Federal do Pará, 2014-07-30) LOBO, Patrícia dos Santos; MASCARENHAS, Joana D'Arc Pereira; http://lattes.cnpq.br/5156164089432435
Acute gastroenteritis (AG) is a major cause of morbidity and mortality of children worldwide, being rotavirus one of the causative agents of AG, responsible for about 38.3% of the cases of hospitalization, resulting in 197,000 deaths of children under five years annually, mostly in developing countries. Rotavirus belongs to the family Reoviridae, genera Rotavirus, have genome composed by 11 segments of double-stranded RNA (dsRNA) and are classified into eight groups/different species (A to H).The most commonly found belongs to the group/species A (RVA), however rotavirus group/species C (RVC) has assumed importance in gastroenteritis and usually relate to self-limiting course of childhood diarrhea, with possible transmission from pigs. This study aims to detect the RVC in children less than three years of age hospitalized with acute gastroenteritis in Belém city, Pará state, Amazon region, Brazil. From May 2008 to April 2011, an intensive surveillance for AG was performed in a pediatric hospital of Belém in children with AG, of which 279 samples had negative results for RVA and norovirus. Accordingly, the nucleic acid was extracted from the fecal suspension with silica by the method of reverse transcribed and amplified by polymerase chain reaction (PCR) using primer pairs specific for the VP7 gene G8S/G8A; C1/C4 for VP6; T434/T435 for VP4; and NSP4 (+)/(-) specific for NSP4 gene. The positive control (prototype Cowden) and negative control (water RNAse free) were used in all tests. Automatic sequencing and subsequent phylogenetic analysis were performed. Data were analyzed using statistical chi-square partition, Fisher's exact test and simple logistic regression with p values ≤ 0.05. The positive rate for RVC was 2.1% (6/279) by PAGE and RT-PCR and posteriorly, submitted to automated sequencing and classified in genotypes I2, G4, P[2] and E2 for VP6, VP7, VP4 and NSP4 genes, respectively. The frequency of RVC was higher in the age group between 24 to 36 months (5.7%) (p= 0.0493). Gender more affected by RVC was the male with a frequency of 83.3%. Signs and/or symptoms most frequent in children with RVC were fever (80%), vomiting (83.3%) and dehydration (100%). In summary, in this study it was possible the characterization and phylogenetic analysis of VP6, VP7, VP4 and NSP4 genes, providing important data regarding the presence of these agents in hospitalized children with gastroenteritis in the metropolitan region of Belém, Pará.