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1. Epidemiology and molecular detection of human adenovirus and non-polio enterovirus in fecal samples of children with acute gastroenteritis: A five-year surveillance in northern Brazil.

Author: Jainara Cristina Dos Santos Alves, Dielle Monteiro Teixeira, Jones Anderson Monteiro Siqueira, Danielle Rodrigues de Deus, Darleise de Souza Oliveira, James Lima Ferreira, Patricia Dos Santos Lobo, Luana da Silva Soares, Fernando Neto Tavares, and Yvone Benchimol Gabbay
Subjects: Medicine, Science
Abstract: Acute gastroenteritis (AGE) is a common pediatric infection that remains a significant cause of childhood morbidity and mortality worldwide, especially in low-income regions. Thus, the objective of this study was to detect human adenovirus (HAdV) and non-polio enterovirus (NPEV) in fecal samples from the Gastroenteritis Surveillance Network, and to identify circulating strains by nucleotide sequencing. A total of 801 fecal samples were tested using qPCR/RT-qPCR, and 657 (82.0%) were inoculated into HEp-2C and RD cell lines. The HAdV and NPEV positivity rates obtained using qPCR/RT-qPCR were 31.7% (254/801) and 10.5% (84/801), respectively, with 5.4% (43/801) co-detection. Cytopathic effect was observed in 9.6% (63/657) of patients, 2.7% (18/657) associated with HAdV, and 6.2% (41/657) associated with NPEV after testing by ICC-PCR. A comparison of the two methodologies demonstrated an agreement of 93.5% for EVNP and 64.4% for HAdV. These two viruses were detected throughout the study period, with HAdV positivity rates ranging from 41% in Amapá to 18% in Pará. The NEPV varied from 18% in Pará/Rondônia to 3% in Acre. The most affected age group was over 60 months for both HAdV and NPEV. Samples previously positive for rotavirus and norovirus, which did not show a major difference in the presence or absence of diarrhea, fever, and vomiting, were excluded from the clinical analyses of these two viruses. These viruses circulated over five years, with a few months of absence, mainly during the months corresponding to the waves of SARS-CoV-2 infection in Brazil. Five HAdV species were identified (A, B, C, D, and F), with a greater predominance of HAdV-F41 (56.5%) followed by HAdV-C (15.2%). Three NPEV species (A, B, and C) were detected, with serotypes E14 (19.3%) and CVA-24 (16.1%) being the most prevalent. The present study revealed a high diversity of NPEV and HAdV types circulating in children with AGE symptoms in the northern region of Brazil.
Published: 2024
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2. Genotype diversity and molecular evolution of noroviruses: A 30-year (1982-2011) comprehensive study with children from Northern Brazil.

Author: Jones Anderson Monteiro Siqueira, Renato da Silva Bandeira, Darleise de Souza Oliveira, Liann Filiphe Pereira Dos Santos, and Yvone Benchimol Gabbay
Subjects: Medicine, Science
Abstract: A chronologically comprehensive 30-year study was conducted that involved children living in Belém, in the Amazon region of Northern Brazil, who participated in eight different studies from October 1982 to April 2011. The children were followed either in the community or in health units and hospitals in order to identify the norovirus genotypes involved in infections during this time. A total of 2,520 fecal specimens were obtained and subjected to RT-PCR and nucleotide sequencing for regions A, B, C, D and P2 of the viral genome. An overall positivity of 16.9% (n = 426) was observed, and 49% of the positive samples were genotyped (208/426), evidencing the presence of several genotypes as follows: Polymerase gene (GI.P4, GII.Pa, GII.Pc, GII.Pe, GII.Pg, GII.Pj, GII.P3, GII.P4, GII.P6, GII.P7, GII.P8, GII.P12, GII.P13, GII.P14, GII.P21, GII.P22), and VP1 gene (GI.3, GI.7, GII.1, GII.2, GII.3, GII.4, GII.6, GII.7, GII.8, GII.10, GII.12, GII.14, GII.17, GII.23). The GII.P4/GII.4 genotype determined by both open reading frames (ORFs) (partial polymerase and VP1 genes) was found for 83 samples, and analyses of the subdomain P2 region showed 10 different variants: CHDC (1970s), Tokyo (1980s), Bristol_1993, US_95/96, Kaiso_2003, Asia_2003, Hunter_2004, Yerseke_2006a, Den Haag_2006b (subcluster "O") and New Orleans_2009. Recombination events were confirmed in 47.6% (n = 20) of the 42 samples with divergent genotyping by ORF1 and ORF2 and with probable different breakpoints within the viral genome. The evolutionary analyses estimated a rate of evolution of 1.02 x 10-2 and 9.05 x 10-3 subs./site/year using regions C and D from the VP1 gene, respectively. The present research shows the broad genetic diversity of the norovirus that infected children for 30 years in Belém. These findings contribute to our understanding of noroviruses molecular epidemiology and viral evolution and provide a baseline for vaccine design.
Published: 2017
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3. Norovirus diversity in diarrheic children from an African-descendant settlement in Belém, Northern Brazil.

Author: Glicélia Cruz Aragão, Joana D'Arc Pereira Mascarenhas, Jane Haruko Lima Kaiano, Maria Silvia Sousa de Lucena, Jones Anderson Monteiro Siqueira, Túlio Machado Fumian, Juliana das Mercês Hernandez, Consuelo Silva de Oliveira, Darleise de Souza Oliveira, Eliete da Cunha Araújo, Luana da Silva Soares, Alexandre Costa Linhares, and Yvone Benchimol Gabbay
Subjects: Medicine, Science
Abstract: Norovirus (NoV), sapovirus (SaV) and human astrovirus (HAstV) are viral pathogens that are associated with outbreaks and sporadic cases of gastroenteritis. However, little is known about the occurrence of these pathogens in relatively isolated communities, such as the remnants of African-descendant villages ("Quilombola"). The objective of this study was the frequency determination of these viruses in children under 10 years, with and without gastroenteritis, from a "Quilombola" Community, Northern Brazil. A total of 159 stool samples were obtained from April/2008 to July/2010 and tested by an enzyme immunoassay (EIA) and reverse transcription-polymerase chain reaction (RT-PCR) to detect NoV, SaV and HAstV, and further molecular characterization was performed. These viruses were detected only in the diarrheic group. NoV was the most frequent viral agent detected (19.7%-16/81), followed by SaV (2.5%-2/81) and HAstV (1.2%-1/81). Of the 16 NoV-positive samples, 14 were sequenced with primers targeting the B region of the polymerase (ORF1) and the D region of the capsid (ORF2). The results showed a broad genetic diversity of NoV, with 12 strains being classified as GII-4 (5-41.7%), GII-6 (3-25%), GII-7 (2-16.7%), GII-17 (1-8.3%) and GI-2 (1-8.3%), as based on the polymerase region; 12 samples were classified, based on the capsid region, as GII-4 (6-50%, being 3-2006b variant and 3-2010 variant), GII-6 (3-25%), GII-17 (2-16.7%) and GII-20 (1-8.3%). One NoV-strain showed dual genotype specificity, based on the polymerase and capsid region (GII-7/GII-20). This study provides, for the first time, epidemiological and molecular information on the circulation of NoV, SaV and HAstV in African-descendant communities in Northern Brazil and identifies NoV genotypes that were different from those detected previously in studies conducted in the urban area of Belém. It remains to be determined why a broader NoV diversity was observed in such a semi-isolated community.
Published: 2013
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4. SAPOVIRUSES IN CHILDREN WITH ACUTE GASTROENTERITIS FROM MANAUS , AMAZON REGION, BRAZIL, 2010-2011

Author: Tammy Kathlyn Amaral REYMÃO, Juliana das Merces HERNANDEZ, Samya Thalita Picanço da COSTA, Maísa Silva de SOUSA, Darleise de Souza OLIVEIRA, Luciana Damascena da SILVA, Renato da Silva BANDEIRA, Ian Carlos Gomes de LIMA, Luana da Silva SOARES, Joana Darc Pereira MASCARENHAS, and Yvone Benchimol GABBAY
Subjects: Sapovirus, Gastroenteritis, Amazon, Arctic medicine. Tropical medicine, RC955-962, Infectious and parasitic diseases, RC109-216
Abstract: SUMMARY Sapoviruses (SaVs) are responsible for acute gastroenteritis in humans, especially children and the elderly. In Brazil, data on SaVs infections are very limited, especially in Northern Brazil. Here, we investigated the occurrence of SaVs in samples from hospitalized children under ten years old that presented acute gastroenteritis. Positive samples were genotyped and phylogenetic analysis was performed using prototype strains sequences obtained from GenBank database. In total, 156 fecal samples were screened by RT-PCR for SaVs. A positivity rate of 3.8% (6/156) was found in children under three years of age. Four genotypes were detected: GI.I, GI.2 and GII.2?-GII.4?/GII.4, suggesting a possible inter-genotypes recombination. Most infections (83.3%) occurred between August and September. The positivity was similar to that found in other countries and genotyping demonstrated the presence of distinct genotypes. To our knowledge, this is the first study reporting the circulation of SaVs in Manaus, state of Amazonas, Amazon region, Brazil.
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5. MOLECULAR EPIDEMIOLOGY TO UNDERSTAND THE SARS-CoV-2 EMERGENCE IN THE BRAZILIAN AMAZON REGION

Author: Mirleide Cordeiro dos Santos, Edivaldo Costa Sousa, Jessylene de Almeida Ferreira, Sandro Patroca da Silva, Michel Platini Caldas de Souza, Jedson Ferreira Cardoso, Amanda Mendes Silva, Luana Soares Barbagelata, Wanderley Dias das Chagas, James Lima Ferreira, Edna Maria Acunã de Souza, Patrícia Louise Araújo Vilaça, Jainara Cristina dos Santos Alves, Michelle Carvalho de Abreu, Patrícia dos Santos Lobo, Fabíolla da Silva dos Santos, Alessandra Alves Polaro Lima, Camila de Marco Bragagnolo, Luana da Silva Soares, Patricía Sousa Moraes de Almeida, Darleise de Souza Oliveira, Carolina Koury Nassar Amorim, Iran Barros Costa, Dielle Monteiro Teixeira, Edvaldo Tavares da Penha, Delana Andreza Melo Bezerra, Jones Anderson Monteiro Siqueira, Fernando Neto Tavares, Felipe Bonfim Freitas, Janete Taynã Nascimento Rodrigues, Janaína Mazaro, Andreia Santos Costa, Márcia Socorro Pereira Cavalcante, Marineide Souza da Silva, Guilherme Alfredo Novelino Araújo, Ilvanete Almeida da Silva, Gleissy Adriane Lima Borges, Lídio Gonçalves de Lima, Hivylla Lorrana dos Santos Ferreira, Miriam Teresinha Furlam Prando Livorati, André Luiz de Abreu, Arnaldo Correia de Medeiros, Hugo Reis Resque, Rita Catarina Medeiros Sousa, and Giselle Maria Rachid Viana
Subjects: Genetic diversity, medicine.medical_specialty, Geography, Phylogenetic tree, Molecular epidemiology, Amazon rainforest, Evolutionary biology, Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), Public health, Pandemic, Epidemiology, medicine
Abstract: The COVID-19 pandemic in Brazil has demonstrated an important public health impact, as has been observed in the world. In Brazil, the Amazon Region contributed with a large number of cases of COVID-19, especially in the beginning of the circulation of SARS-CoV-2 in the country. Thus, we describe the epidemiological profile of COVID-19 and the genetic diversity of SARS-CoV-2 strains circulating in the Amazon Region. We observe an extensive spread of virus in this Brazilian site. The data on sex, age and symptoms presented by the investigated individuals were similar to what has been observed worldwide. The genomic analysis of the viruses revealed important amino acid changes, including the D614G and the I33T in Spike and ORF6 proteins, respectively. The latter found in strains originating in Brazil. The phylogenetic analyzes demonstrated the circulation of the lineages B.1 and B.1.1, whose circulation in Brazil has already been previous reported. Our data reveals molecular epidemiology of SARS-CoV-2 in the Amazon Region. These findings also reinforce the importance of continuous genomic surveillance this virus with the aim of providing accurate and updated data to understand and map the transmission network of this agent in order to subsidize operational decisions in public health.
Published: 2020
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6. Avaliação diagnóstica das infecções por vírus Epstein-Barr, parvovírus B19 e vírus linfotrópico de células T humanas em pacientes portadores de lúpus eritematoso sistêmico em hospital de referência do Estado do Pará, Brasil

Author: Olinda Macêdo, Kelly Beatriz Romeiro Araújo, Iran Barros Costa, Alessandra Alves Polaro, Carolina Barros Kahwage, Wellington Teixeira Viana Júnior, Bruna Ramos dos Santos, Darleise de Souza Oliveira, Larissa Teixeira Meireles, Francinilza Magno Silva, Felipe Bonfim Freitas, Igor Brasil-Costa, Andreza Paloma Góes Oliveira, Talita Antônia Furtado Monteiro, Juliana Marinho Melo, Antônio de Moura, Mayara Jane Miranda da Silva, and Manoel Gomes da Silva Filho
Subjects: Strategy and Management, Mechanical Engineering, Metals and Alloys, Industrial and Manufacturing Engineering
Abstract: El lupus eritematoso sistemico (LES) es una enfermedad autoinmune cronica, cuyo desarrollo puede estar asociado a la infeccion por virus, como el virus de Epstein-Barr (VEB), parvovirus B19 y virus linfotropico de celulas T humanas (HTLV). Durante el periodo de junio a setiembre de 2014, se realizo un estudio transversal, incluyendo a 85 pacientes oriundos del Hospital Jean Bitar en la Ciudad de Belem, Estado de Para, Brasil. Se hizo el estudio de anticuerpos especificos contra los agentes virales estudiados, ademas de investigar la presencia del genoma para EBV y HTLV. Se evaluaron tambien, variables clinicas y epidemiologicas. La mayoria de los pacientes era de mujeres, con un promedio de edad de 30 anos y se declararon blancos. Para VEB, se detecto una positividad de 37,6% para IgM, 98,8% para IgG y 2,4% por PCR, con cuantificaciones de 85.028 y 298 copias del genoma/mL de plasma. Para B19, la positividad para IgM fue 0% y para IgG 67,1%. No hubo deteccion serologica o por PCR de HTLV. Se verifico una relacion estadisticamente significante entre la positividad para IgM anti VEB y pacientes mas jovenes. Este hallazgo puede estar relacionado con la mayor eficiencia en la produccion de esa inmunoglobulina en las infecciones primarias agudas, que ocurren generalmente en individuos infantes o adultos jovenes. Adicionalmente, el resultado de IgG anti B19 fue asociado a la edad avanzada de los pacientes, probablemente por un tiempo mayor de exposicion al virus aliado a la persistencia de este marcador luego del contacto. La presencia de los marcadores no estuvo asociada a las variables clinicas y epidemiologicas. Sin embargo, el porcentual de positividad para el marcador de infeccion aguda por VEB puede sugerir que haya una implicacion del virus con el LES
Published: 2016
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7. Norovirus genogroups I and II in environmental water samples from Belém city, Northern Brazil

Author: P. K. P Spada, Lena Líllian Canto de Sá Morais, Tulio Machado Fumian, Yvone Benchimol Gabbay, Dielle Monteiro Teixeira, Darleise de Souza Oliveira, Ian Carlos Gomes de Lima, Renato da Silva Bandeira, Maísa Silva de Sousa, Tereza Cristina Monteiro Gurjão, and Joana D'Arc Pereira Mascarenhas
Subjects: 0301 basic medicine, Microbiology (medical), Veterinary medicine, Genotype, 030231 tropical medicine, 030106 microbiology, Sewage, Fresh Water, Biology, Real-Time Polymerase Chain Reaction, medicine.disease_cause, Microbiology, Open Reading Frames, 03 medical and health sciences, 0302 clinical medicine, Environmental water, medicine, Waste Management and Disposal, Effluent, Phylogeny, Water Science and Technology, Reverse Transcriptase Polymerase Chain Reaction, Sequence Analysis, RNA, business.industry, Aquatic ecosystem, Norovirus, Public Health, Environmental and Occupational Health, Reverse transcription polymerase chain reaction, Infectious Diseases, business, Brazil, Environmental Monitoring
Abstract: This study investigated the presence of norovirus (NoV) GI and GII in environmental samples from the northern region of Brazil. Water samples were collected monthly (November 2008/October 2010) from different sources and sewage and concentrated by the adsorption-elution method. The NoV investigation used molecular methods followed by sequencing reactions. The general positivity for NoV was 33.9% (57/168). Considering the results obtained only in the semi-nested RT-PCR (reverse transcription polymerase chain reaction) and only in the TaqMan® real-time PCR, the rates were 26.8% (45/168) and 27.4% (46/168), respectively, being for NoV GI 22.2% (10/45) and 19.6% (9/46); for GII 17.8% (8/45) and 15.2% (7/46); and for GI + GII 60% (27/45) and 65.2% (30/46), respectively. Different GI (GI.1, GI.4, GI.7 and GI.8) and GII (GII.4, GII.6, GII.9, GII.12 and GII.14) genotypes were detected. These results demonstrated the NoV was disseminated in the waters of Belém city due to a lack of sanitation that allowed the discharge of contaminated effluents into these aquatic ecosystems.
Published: 2016
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8. Epidemiological and molecular investigation of norovirus and astrovirus infections in Rio Branco, Acre, Northern Brazil: A retrospective study

Author: Yvone Benchimol Gabbay, Dielle Monteiro Teixeira, Luana da Silva Soares, Luciana Damascena da Silva, Evandro Leite Rodrigues Bitencurt, Mônica Cristina de Moraes Silva, Renato da Silva Bandeira, Edvaldo Carlos Brito Loureiro, Tallyta Barros Medeiros, Ricardo José de Paula Souza e Guimarães, Rita S.U. Silva, Joana D'Arc Pereira Macarenhas, Darleise de Souza Oliveira, and Jones Anderson Monteiro Siqueira
Subjects: Male, Veterinary medicine, viruses, Norovirus / patogenicidade, medicine.disease_cause, Feces, 0302 clinical medicine, Infec??es por Caliciviridae, Astroviridae Infections, Epidemiology, Prevalence, Infec??es por Astroviridae, 030212 general & internal medicine, Child, Phylogeny, Caliciviridae Infections, biology, Fezes / virologia, Incidence (epidemiology), virus diseases, Avastrovirus / patogenicidade, Middle Aged, Gastroenterite / virologia, Gastroenteritis, Diarrhea, Infectious Diseases, Child, Preschool, RNA, Viral, 030211 gastroenterology & hepatology, Female, medicine.symptom, Estudos Transversais / m?todos, Brazil, Adult, medicine.medical_specialty, Adolescent, Genotype, Astrovirus, 03 medical and health sciences, Young Adult, Rio Branco (AC), Virology, medicine, Humans, Retrospective Studies, Varia??o Gen?tica, business.industry, Norovirus, Infant, Newborn, Outbreak, Genetic Variation, Infant, Retrospective cohort study, biology.organism_classification, Astroviridae, business
Abstract: University of the State of Par?. Postgraduate Program in Parasitic Biology in the Amazon. Bel?m, PA, Brazil. Minist?rio da Sa?de. Secretaria de Vigil?ncia em Sa?de. Instituto Evandro Chagas. Ananindeua, PA, Brasil. Minist?rio da Sa?de. Secretaria de Vigil?ncia em Sa?de. Instituto Evandro Chagas. Ananindeua, PA, Brasil. Minist?rio da Sa?de. Secretaria de Vigil?ncia em Sa?de. Instituto Evandro Chagas. Ananindeua, PA, Brasil. Minist?rio da Sa?de. Secretaria de Vigil?ncia em Sa?de. Instituto Evandro Chagas. Ananindeua, PA, Brasil. Minist?rio da Sa?de. Secretaria de Vigil?ncia em Sa?de. Instituto Evandro Chagas. Ananindeua, PA, Brasil. Minist?rio da Sa?de. Secretaria de Vigil?ncia em Sa?de. Instituto Evandro Chagas. Ananindeua, PA, Brasil. Minist?rio da Sa?de. Secretaria de Vigil?ncia em Sa?de. Instituto Evandro Chagas. Ananindeua, PA, Brasil. Minist?rio da Sa?de. Secretaria de Vigil?ncia em Sa?de. Instituto Evandro Chagas. Ananindeua, PA, Brasil. Municipality Secretary of Health of Rio Branco. Acre, AC, Brazil. Minist?rio da Sa?de. Secretaria de Vigil?ncia em Sa?de. Instituto Evandro Chagas. Ananindeua, PA, Brasil. Minist?rio da Sa?de. Secretaria de Vigil?ncia em Sa?de. Instituto Evandro Chagas. Ananindeua, PA, Brasil. Minist?rio da Sa?de. Secretaria de Vigil?ncia em Sa?de. Instituto Evandro Chagas. Ananindeua, PA, Brasil. Minist?rio da Sa?de. Secretaria de Vigil?ncia em Sa?de. Instituto Evandro Chagas. Ananindeua, PA, Brasil. Norovirus (NoV) is a major cause of nonbacterial acute gastroenteritis (AGE) outbreaks worldwide, with infections reported in semiclosed environments, particularly in hospitals and nursing homes. Astrovirus (HAstV) is prevalent worldwide, especially in developing countries. We aimed to determine the prevalence, spatial distribution, and genetic diversity of NoV and HAstV in children under 5 years of age in Rio Branco city, Acre State, Amazon Region, Brazil. Stool samples from children with (n?=?240) and without (n?=?248) AGE were collected from January to December 2012 from seven neighborhoods. The overall NoV prevalence was 12.3% (60 of 488); representing 15.8% (38 of 240) of the symptomatic samples and 8.9% (22 of 248) of the controls. HAstVs infection was observed in 4.7% (23 of 488) of the samples tested, 6.2% (15 of 240) of AGE cases, and 2.4% (6 of 248) of the controls (plus two without information about feces consistency). Infections were found in all age groups with higher frequency in children less than two years of age, for both viruses. NoV was detected in all neighborhoods, with a higher concentration in the fourth (30%; 18 of 60). NoV nucleotide sequencing performed in 86.7% (52 of 60) of the positive samples showed the circulation of the strains GII.4 (57.7%; 30 of 52), GIIPe/GII.4 (19.2%; 10 of 52), GII.7, GII.Pg/GII.1, and GII.Pc (3.8%; 2 of 52 for each), GII.6 and GII.Pg (1.9%; 1 of 52 for each), and GI.3 (7.7%; 4 of 52). Three GII.4 variants were detected: Den Haag_2006b (n?=?1), New Orleans_2009 (n?=?1), and Sydney_2012 (n?=?14). HAstV types HAstV-1a (81.8%; 9 of 11) and HAstV-2c (18.2%; 2 of 11) were observed in the 47.8% (11 of 23) of characterized samples. This is the first data obtained in Acre State regarding the prevalence of these viruses and provides epidemiological and molecular information for a better understanding of their role among children with and without AGE.
Published: 2018

9. Occurrence of Norovirus GIV in Environmental Water Samples from Belém City, Amazon Region, Brazil

Author: Joana D'Arc Pereira Mascarenhas, Dielle Monteiro Teixeira, Tereza Cristina Monteiro Gurjão, Darleise de Souza Oliveira, Yvone Benchimol Gabbay, Juliana das Merces Hernandez, Luciana Damascena da Silva, Lena Líllian Canto de Sá Morais, P. K. P Spada, and Alexandre da Costa Linhares
Subjects: 0301 basic medicine, Veterinary medicine, Genotype, Epidemiology, viruses, Health, Toxicology and Mutagenesis, 030106 microbiology, Sewage, Norovirus / isolamento & purifica??o, medicine.disease_cause, 03 medical and health sciences, fluids and secretions, Rivers, Norovirus GIV, Environmental water, Virology, medicine, Humans, ?guas de Superf?cie, Caliciviridae Infections, business.industry, Amazon rainforest, Norovirus, Gastroenterite, virus diseases, Outbreak, digestive system diseases, Gastroenterite / virologia, Esgotos / virologia, Gastroenteritis, Geography, Norovirus / gen?tica, ?gua Doce, Rea??o em Cadeia da Polimerase Via Transcriptase Reversa / m?todos, business, Surface water, Bay, Brazil, Food Science
Abstract: Minist?rio da Sa?de. Secretaria de Vigil?ncia em Sa?de. Instituto Evandro Chagas. Ananindeua, PA, Brasil / Federal University of Para. Tropical Medicine Center. Postgraduate Program in Tropical Diseases. Bel?m, PA, Brazil. Minist?rio da Sa?de. Secretaria de Vigil?ncia em Sa?de. Instituto Evandro Chagas. Ananindeua, PA, Brasil. Minist?rio da Sa?de. Secretaria de Vigil?ncia em Sa?de. Instituto Evandro Chagas. Ananindeua, PA, Brasil. Minist?rio da Sa?de. Secretaria de Vigil?ncia em Sa?de. Instituto Evandro Chagas. Ananindeua, PA, Brasil. Minist?rio da Sa?de. Secretaria de Vigil?ncia em Sa?de. Instituto Evandro Chagas. Ananindeua, PA, Brasil. Minist?rio da Sa?de. Secretaria de Vigil?ncia em Sa?de. Instituto Evandro Chagas. Ananindeua, PA, Brasil. Minist?rio da Sa?de. Secretaria de Vigil?ncia em Sa?de. Instituto Evandro Chagas. Ananindeua, PA, Brasil. Minist?rio da Sa?de. Secretaria de Vigil?ncia em Sa?de. Instituto Evandro Chagas. Ananindeua, PA, Brasil. Minist?rio da Sa?de. Secretaria de Vigil?ncia em Sa?de. Instituto Evandro Chagas. Ananindeua, PA, Brasil. Minist?rio da Sa?de. Secretaria de Vigil?ncia em Sa?de. Instituto Evandro Chagas. Ananindeua, PA, Brasil. Noroviruses are the major cause of non-bacterial acute gastroenteritis outbreaks in humans, with few reports about the occurrence of the norovirus GIV strain. We investigated the presence of norovirus GIV in surface water (river, bay, and stream) and untreated sewage, and we determined a positivity rate of 9.4 % (9/96). The strains genotyped were GIV.1. To our knowledge, this is the first report of GIV in Brazil.
Published: 2015
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10. Prevalence and molecular diversity of adenovirus in children with severe acute gastroenteritis, in Bel?m, Par? State, before and after the introduction of the rotavirus vaccine in Brazil

Author: Elza Caroline Alves Müller, Maísa Silva de Sousa, Luana da Silva Soares, Darleise de Souza Oliveira, and Alexandre da Costa Linhares
Subjects: Gastroenterite / epidemiologia, Fezes / virologia, Strategy and Management, Mechanical Engineering, Metals and Alloys, Infec??es por Adenovirus Humanos / epidemiologia, Vacinas contra Rotavirus, Infec??es por Adenovirus Humanos / etiologia, T?cnicas Imunoenzim?ticas / m?todos, Industrial and Manufacturing Engineering, Diarreia Infantil / preven??o & controle, Diarreia / epidemiologia, Sequ?ncia de Bases / gen?tica, Adenov?rus Humanos / classifica??o, Epidemiologia Molecular, Infec??es por Adenovirus Humanos / preven??o & controle, Rea??o em Cadeia da Polimerase / m?todos, Imunocromatografia / m?todos, DNA Viral / isolamento & purifica??o
Abstract: Minist?rio da Sa?de. Secretaria de Vigil?ncia em Sa?de. Instituto Evandro Chagas. Ananindeua, PA, Brasil / Universidade Federal do Par?. Programa de P?s-Gradua??o em Doen?as Tropicais. Bel?m, PA, Brasil / Universidade Federal do Amap?. Coordena??o de Farm?cia. Macap?, AP, Brasil. Minist?rio da Sa?de. Secretaria de Vigil?ncia em Sa?de. Instituto Evandro Chagas. Ananindeua, PA, Brasil Minist?rio da Sa?de. Secretaria de Vigil?ncia em Sa?de. Instituto Evandro Chagas. Ananindeua, PA, Brasil Minist?rio da Sa?de. Secretaria de Vigil?ncia em Sa?de. Instituto Evandro Chagas. Ananindeua, PA, Brasil Universidade Federal do Par?. Programa de P?s-Gradua??o em Doen?as Tropicais. Bel?m, PA, Brasil. OBJETIVOS: Detectar a presen?a e definir os tipos de adenov?rus humanos (HAdV) em indiv?duos menores de 3 anos de idade, participantes de dois projetos de pesquisa sobre gastroenterites virais, realizados em Bel?m, estado do Par?, Brasil: o primeiro, um estudo de vigil?ncia hospitalar e ambulatorial, pr?-vacina??o contra rotav?rus (RV), realizado pelo Instituto Evandro Chagas no per?odo de mar?o a setembro de 2003; e o segundo, o projeto "Rotav?rus Caso-Controle", p?s-vacina??o, com crian?as hospitalizadas apresentando quadro de gastroenterite aguda no per?odo de maio de 2008 a abril de 2009. MATERIAIS E M?TODOS: Foram testadas amostras fecais de 1.160 crian?as, utilizando-se as t?cnicas de ensaio imunoenzim?tico e imunocromatografia para triagem, a rea??o em cadeia da polimerase e sequenciamento de nucleot?deos para tipagem e identifica??o molecular. RESULTADOS: Os HAdV foram encontrados em 7,2% (84/1.160) das amostras e os adenov?rus ent?ricos em 5,0% (58/1.160), equivalendo a 69,0% (58/84) dos casos positivos. Desses, 3,3% (25/760) eram do estudo pr?-vacina??o contra RV e 8,2% (33/400) do p?s-vacina??o. A rea??o de sequenciamento caracterizou a esp?cie F como a mais prevalente na regi?o, equivalendo a 70,0% (21/30), com o tipo 41 encontrado em 85,7% (18/21) dos casos positivos. CONCLUS?O: Esses resultados evidenciaram a circula??o de HAdV entre a popula??o infantil de Bel?m, demonstrando a sua import?ncia como causa de gastroenterite com ou sem hospitaliza??o, tanto antes como ap?s a introdu??o da vacina contra RV na rede p?blica de sa?de no Brasil. OBJECTIVES: To detect the presence and to define the human adenovirus (HAdV) types from individuals under 3 years old in two research projects on viral gastroenteritis in Bel?m, Par? State, Brazil: the first, an inpatient and outpatient surveillance study on rotavirus (RV) pre-vaccination performed by Instituto Evandro Chagas from March to September 2003; and the second, from the "Rotavirus Case-Control" (post vaccination) study, carried out from May 2008 to April 2009, with hospitalized children due to acute gastroenteritis. MATERIALS AND METHODS: Faecal samples were tested from 1,160 children using immunoenzymatic assay and immunochromatography techniques for screening, polymerase chain reaction and nucleotide sequencing for typing and molecular identification. RESULTS: HAdV were found in 7.2% (84/1,160) of the samples, and enteric adenoviruses in 5.0% (58/1,160), equivalent to 69.0% (58/84) of the positive cases. Of these, 3.3% (25/760) were from the RV pre-vaccination study and 8.2% (33/400) from the post-vaccination. The sequencing reaction characterized the F specie as the most prevalent in the region, equivalent to 70.0% (21/30), with the type 41 in 85.7% (18/21) of the positive cases. CONCLUSION: These results showed HAdV circulation among the infant population in Bel?m, demonstrating its importance as gastroenteritis agent either hospitalized or not, both before and after the RV vaccine introduction on the Brazilian public health network.
Published: 2017

11. Genotype diversity and molecular evolution of noroviruses: a 30-year (1982-2011) comprehensive study with children from Northern Brazil

Author: Yvone Benchimol Gabbay, Liann Filiphe Pereira dos Santos, Darleise de Souza Oliveira, Renato da Silva Bandeira, and Jones Anderson Monteiro Siqueira
Subjects: Artificial Gene Amplification and Extension, Norovirus / isolamento & purifica??o, Pathology and Laboratory Medicine, Biochemistry, Polymerases, Polymerase Chain Reaction, Geographical locations, DNA Complementar / gen?tica, lcsh:Science, Child, Phylogeny, Caliciviridae Infections, Recombination, Genetic, RNA Viral / gen?tica, virus diseases, Medical Microbiology, Viral Pathogens, Child, Preschool, An?lise de Sequ?ncia de DNA, Ensaio de Imunoadsor??o Enzim?tica / m?todos, Genotype, Bioinformatics, Microbiology, Evolution, Molecular, 03 medical and health sciences, Gen?tipo, Molecular evolution, DNA-binding proteins, Humans, Molecular Biology Techniques, Genotyping, Microbial Pathogens, Molecular Biology, Molecular epidemiology, Genoma Viral / gen?tica, lcsh:R, Organisms, Proteins, Computational Biology, Genetic Variation, Reverse Transcriptase-Polymerase Chain Reaction, History, 20th Century, Virology, 030104 developmental biology, RNA Viral / isolamento & purifica??o, Varia??o Gen?tica / gen?tica, lcsh:Q, People and places, Recombina??o Gen?tica, 0301 basic medicine, RNA viruses, Evolutionary Genetics, viruses, lcsh:Medicine, medicine.disease_cause, Viral Packaging, Database and Informatics Methods, Sequenciamento de Nucleot?deos em Larga Escala, fluids and secretions, Medicine and Health Sciences, Genetics, Multidisciplinary, Fezes / virologia, Gastroenteritis, Viral evolution, Viruses, Rea??o em Cadeia da Polimerase Via Transcriptase Reversa / m?todos, RNA, Viral, Pathogens, Sequence Analysis, Brazil, Research Article, Taxa de Muta??o, Genome, Viral, Biology, Research and Analysis Methods, History, 21st Century, Viral Evolution, Caliciviruses, Evolu??o Molecular, Epidemiologia Molecular, Genetic variation, medicine, Genetic diversity, Evolutionary Biology, Biology and life sciences, Norovirus, South America, Organismal Evolution, Viral Replication, Filogenia, Norovirus / gen?tica, Microbial Evolution, Distribui??o Temporal, Rea??o em Cadeia da Polimerase em Tempo Real / m?todos
Abstract: This research was supported by Evandro Chagas Institute and Funda??o Amaz?nia de Amparo a Estudos e Pesquisa do Par? ? Fapespa (Edital: 006/2014 ? Programa de aux?lio ? pesquisa/Projetos tem?ticos da Fapespa: Sa?de). Minist?rio da Sa?de. Secretaria de Vigil?ncia em Sa?de. Instituto Evandro Chagas. Laborat?rio de Norov?rus e outros V?rus Gastroent?ricos. Ananindeua, PA, Brasil. Minist?rio da Sa?de. Secretaria de Vigil?ncia em Sa?de. Instituto Evandro Chagas. Ananindeua, PA, Brasil. Minist?rio da Sa?de. Secretaria de Vigil?ncia em Sa?de. Instituto Evandro Chagas. Ananindeua, PA, Brasil. Minist?rio da Sa?de. Secretaria de Vigil?ncia em Sa?de. Instituto Evandro Chagas. Laborat?rio de Norov?rus e outros V?rus Gastroent?ricos. Ananindeua, PA, Brasil. Minist?rio da Sa?de. Secretaria de Vigil?ncia em Sa?de. Instituto Evandro Chagas. Laborat?rio de Norov?rus e outros V?rus Gastroent?ricos. Ananindeua, PA, Brasil. A chronologically comprehensive 30-year study was conducted that involved children living in Bel?m, in the Amazon region of Northern Brazil, who participated in eight different studies from October 1982 to April 2011. The children were followed either in the community or in health units and hospitals in order to identify the norovirus genotypes involved in infections during this time. A total of 2,520 fecal specimens were obtained and subjected to RT-PCR and nucleotide sequencing for regions A, B, C, D and P2 of the viral genome. An overall positivity of 16.9% (n = 426) was observed, and 49% of the positive samples were genotyped (208/426), evidencing the presence of several genotypes as follows: Polymerase gene (GI.P4, GII.Pa, GII.Pc, GII.Pe, GII.Pg, GII.Pj, GII.P3, GII.P4, GII.P6, GII.P7, GII.P8, GII.P12, GII.P13, GII.P14, GII.P21, GII.P22), and VP1 gene (GI.3, GI.7, GII.1, GII.2, GII.3, GII.4, GII.6, GII.7, GII.8, GII.10, GII.12, GII.14, GII.17, GII.23). The GII.P4/GII.4 genotype determined by both open reading frames (ORFs) (partial polymerase and VP1 genes) was found for 83 samples, and analyses of the subdomain P2 region showed 10 different variants: CHDC (1970s), Tokyo (1980s), Bristol_1993, US_95/96, Kaiso_2003, Asia_2003, Hunter_2004, Yerseke_2006a, Den Haag_2006b (subcluster ?O?) and New Orleans_2009. Recombination events were confirmed in 47.6% (n = 20) of the 42 samples with divergent genotyping by ORF1 and ORF2 and with probable different breakpoints within the viral genome. The evolutionary analyses estimated a rate of evolution of 1.02 x 10?2 and 9.05 x 10?3 subs./site/year using regions C and D from the VP1 gene, respectively. The present research shows the broad genetic diversity of the norovirus that infected children for 30 years in Bel?m. These findings contribute to our understanding of noroviruses molecular epidemiology and viral evolution and provide a baseline for vaccine design.
Published: 2017

12. Astrov?rus infection in hospitalized children: molecular, clinical and epidemiological features

Author: Yvone Benchimol Gabbay, Thayara Morais Portal, Maria Cleonice Aguiar Justino, Darleise de Souza Oliveira, Thaís Cristina Nascimento de Carvalho, Hugo Reis Resque, Jones Anderson Monteiro Siqueira, and Luciana Damascena da Silva
Subjects: 0301 basic medicine, medicine.medical_specialty, 030106 microbiology, medicine.disease_cause, Astrovirus, Feces, 03 medical and health sciences, Astroviridae Infections, Virology, Internal medicine, Genotype, Epidemiology, Prevalence, medicine, Humans, Diarreia, Genotyping, Phylogeny, Molecular Epidemiology, biology, business.industry, Mamastrovirus / patogenicidade, Infant, Newborn, Gastroenterite, Genetic Variation, Infant, Outbreak, Bel?m (PA), biology.organism_classification, Astroviridae / patogenicidade, Regi?o Amaz?nica (BR), Gastroenteritis, Hospitalization, Diarrhea, 030104 developmental biology, Infectious Diseases, Child, Preschool, Norovirus, Astroviridae, medicine.symptom, business, Viral load, Brazil
Abstract: Minist?rio da Sa?de. Secretaria de Vigil?ncia em Sa?de. Instituto Evandro Chagas. Ananindeua, PA, Brasil. Minist?rio da Sa?de. Secretaria de Vigil?ncia em Sa?de. Instituto Evandro Chagas. Ananindeua, PA, Brasil. Minist?rio da Sa?de. Secretaria de Vigil?ncia em Sa?de. Instituto Evandro Chagas. Programa de P?s-Gradua??o em Virologia. Ananindeua, PA, Brasil. Minist?rio da Sa?de. Secretaria de Vigil?ncia em Sa?de. Instituto Evandro Chagas. Programa de P?s-Gradua??o em Virologia. Ananindeua, PA, Brasil. Minist?rio da Sa?de. Secretaria de Vigil?ncia em Sa?de. Instituto Evandro Chagas. Ananindeua, PA, Brasil. Minist?rio da Sa?de. Secretaria de Vigil?ncia em Sa?de. Instituto Evandro Chagas. Ananindeua, PA, Brasil. Minist?rio da Sa?de. Secretaria de Vigil?ncia em Sa?de. Instituto Evandro Chagas. Ananindeua, PA, Brasil. Minist?rio da Sa?de. Secretaria de Vigil?ncia em Sa?de. Instituto Evandro Chagas. Ananindeua, PA, Brasil. Background: Astrovirus (HAstV) is a common viral pathogen that causes gastroenteritis worldwide. It is classified into eight classical human types (HAstV-1/8) and seven other less prevalent types, described as HAstV VA1, VA2, VA3, VA4, MLB-1, MLB-2 and MLB-3. During outbreaks, the elderly and children are the most affected, and the spread of the virus is associated with person-to-person contact, food ingestion and contaminated water. Objectives: The aim of the present study was to investigate the prevalence of infection and genetic diversity of HAstV strains. Samples were collected from children with acute gastroenteritis admitted to a large pediatric hospital during a surveillance period of three years (2008?2011) in Bel?m city, Par? State, Amazon Region, Northern Brazil. Study design: Screening and genotyping tests were conducted using RT-PCR to detect the classical and non-classical HAstV types using specific primers. A semi-nested RT-PCR protocol was developed to improve viral detection in samples with a low viral load. Results: The overall positivity observed in this study was 3.9% (19/483). The age distribution showed a high prevalence of positive cases in children under one year old (5.3%). We found vomiting associated with 75% of the positive cases, fever with 82.3%, and dehydration with 76.9%. Most patients with positive cases demonstrated two to five days of diarrhea, two to three episodes of vomiting during hospitalization, and three bowel movements per day. Co-infection with HAstV and norovirus was observed in three cases (15.8%), and no pattern of seasonality or any relationship between the HAstV positivity rate and climate variables was observed. Eighteen positive samples (94.7%?18/19) were genotyped based on the ORF 2 region, and the greatest prevalence was of HAstV-1a (66.6%?12/18), followed by HAstV-2 (22.2%?4/18, comprising two type-2b and two type-2c genotypes), HAstV-3c (5.6%?1/18) and HAstV-4c (5.6%?1/18). No non-classical types were detected in the clinical samples analyzed. Conclusions: The present study showed that although HAstV infections occur at low frequency, they are involved in severe pediatric cases of acute gastroenteritis presenting with a high diversity of strains, including the lineages 3c and 4c, which were never before detected in Brazil.
Published: 2017

13. Detection, epidemiology and characterization of VP6 and VP7 genes of group D rotavirus in broiler chickens

Author: Yvone Benchimol Gabbay, Jane Haruko Lima Kaiano, Darleise de Souza Oliveira, Alexandre da Costa Linhares, René Ribeiro da Silva, Joana D'Arc Pereira Mascarenhas, and Delana Andreza Melo Bezerra
Subjects: Rotavirus, medicine.medical_specialty, Veterinary medicine, Componentes Gen?micos / gen?tica, Sequence analysis, Molecular Sequence Data, Nucleotide sequencing, Genome, Viral, Biology, Genome, Rotavirus Infections, Feces, Food Animals, Epidemiology, medicine, Animals, Rea??o em Cadeia da Polimerase / m?todos, Amino Acid Sequence, Antigens, Viral, Gene, Phylogeny, RNA, Double-Stranded, Base Sequence, General Immunology and Microbiology, Reverse Transcriptase Polymerase Chain Reaction, business.industry, Broiler, virus diseases, Sequence Analysis, DNA, Poultry farming, Virology, Group D rotavirus, RNA, Viral, Capsid Proteins, Animal Science and Zoology, business, Chickens, Sequence Alignment, Brazil
Abstract: Minist?rio da Sa?de. Secretaria de Vigil?ncia em Sa?de. Instituto Evandro Chagas. Ananindeua, PA, Brasil Minist?rio da Agricultura. Laborat?rio Nacional Agropecu?rio. Bel?m, PA, Brazil. Minist?rio da Sa?de. Secretaria de Vigil?ncia em Sa?de. Instituto Evandro Chagas. Ananindeua, PA, Brasil. Minist?rio da Sa?de. Secretaria de Vigil?ncia em Sa?de. Instituto Evandro Chagas. Ananindeua, PA, Brasil. Minist?rio da Sa?de. Secretaria de Vigil?ncia em Sa?de. Instituto Evandro Chagas. Ananindeua, PA, Brasil. Minist?rio da Sa?de. Secretaria de Vigil?ncia em Sa?de. Instituto Evandro Chagas. Ananindeua, PA, Brasil. Minist?rio da Sa?de. Secretaria de Vigil?ncia em Sa?de. Instituto Evandro Chagas. Ananindeua, PA, Brasil. Rotaviruses infect humans and animals and are classified into eight groups (A to H). Group D rotavirus (RVD) has been described in birds, although relatively few reports are available. The present study focused on RVD, including epidemiological and molecular aspects of samples collected from broiler chickens in the state of Par?, Brazil. A total of 85 faecal samples were collected between 2008 and 2011 from 37 chicken farms located in eight different municipalities. The viral double-stranded RNA was extracted from faecal suspensions and analysed using polyacrylamide gel electrophoresis (PAGE), followed by reverse transcriptase-polymerase chain reaction (RTPCR) and nucleotide sequencing of the VP6 and VP7 genes. Comparing the positive results, 16.5 per cent (14/85) were obtained by PAGE and 35.3 per cent (30/85) by RT-PCR. Samples from seven of eight municipalities were positive for RVD and infections were recorded in 17 (45.9 per cent) of 37 chicken farms. The RVD infection rate was significantly higher in the 16-day to 30-day age group (62.2 per cent; 23/37) compared with other ages. No consistent relationship was found between the infection rate and either the population density in poultry houses or the climatic conditions. The nucleotide sequences of the VP6 gene were 89.9 to 90.9 per cent similar to the prototype strain 05V0049 and were 88.3 to 100 per cent similar among themselves; VP7 gene nucleotide sequences were 84.3 to 85.4 per cent similar to the prototype strain 05V0049 and 93.8 to 100 per cent similar among themselves. Overall, this study provides new insights into the epidemiology and genome characterization of group D rotaviruses
Published: 2014
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14. Aislamiento e identificaci?n molecular de los enterovirus no polio en casos de par?lisis fl?ccida aguda, ocurridos en la Regi?n Norte de Brasil, en el per?odo de 1996 a 2006

Author: Ana Lucia Monteiro Wanzeller, Edna da Silveira, Ceyla Maria Oeiras de Castro, Euda Galiza Primo, Maria de Lourdes Contente Gomes, Alexandre da Costa Linhares, Jainara Cristina dos Santos Alves, Darleise de Souza Oliveira, and Antonia dos Santos Alves
Subjects: Enterovirus Humano B, Infec??es por Coxsackievirus / epidemiologia, Strategy and Management, Mechanical Engineering, Rea??o em Cadeia da Polimerase Via Transcriptase Reversa / m?todos, Metals and Alloys, Enterovirus / isolamento & purifica??o, Industrial and Manufacturing Engineering
Abstract: Instituto Evandro Chagas/SVS/MS e ao Programa Institucional de Inicia??o Cient?fica (PIBIC-IEC) do Conselho Nacional de Desenvolvimento Cient?fico e Tecnol?gico (CNPq) e Funda??o Amaz?nia Paraense de Amparo ? Pesquisa (FAPESPA). Minist?rio da Sa?de. Secretaria de Vigil?ncia em Sa?de. Instituto Evandro Chagas. Ananindeua, PA, Brasil. Minist?rio da Sa?de. Secretaria de Vigil?ncia em Sa?de. Instituto Evandro Chagas. Ananindeua, PA, Brasil. Minist?rio da Sa?de. Secretaria de Vigil?ncia em Sa?de. Instituto Evandro Chagas. Ananindeua, PA, Brasil. Minist?rio da Sa?de. Secretaria de Vigil?ncia em Sa?de. Instituto Evandro Chagas. Ananindeua, PA, Brasil. Minist?rio da Sa?de. Secretaria de Vigil?ncia em Sa?de. Instituto Evandro Chagas. Ananindeua, PA, Brasil. Minist?rio da Sa?de. Secretaria de Vigil?ncia em Sa?de. Instituto Evandro Chagas. Ananindeua, PA, Brasil. Minist?rio da Sa?de. Secretaria de Vigil?ncia em Sa?de. Instituto Evandro Chagas. Ananindeua, PA, Brasil. Minist?rio da Sa?de. Secretaria de Vigil?ncia em Sa?de. Instituto Evandro Chagas. Ananindeua, PA, Brasil. Minist?rio da Sa?de. Secretaria de Vigil?ncia em Sa?de. Instituto Evandro Chagas. Ananindeua, PA, Brasil. Os enterov?rus (EV) de origem humana pertencem ? fam?lia Picornaviridae, possuem 30 nm de di?metro, genoma de RNA de fita simples, sua transmiss?o ? de via fecal-oral e t?m sido relacionados a surtos de diversas doen?as do sistema nervoso central. O estudo objetivou identificar a presen?a de enterov?rus n?o p?lio (EVNP) isolados de casos de paralisia fl?cida aguda (PFA). Linhagens celulares HEp-2C e RD que apresentaram efeito citop?tico foram selecionadas para testes moleculares posteriores. O RNA viral foi extra?do utilizando-se o kit QIAamp Viral RNA. Os iniciadores 222 e 292, os quais amplificam parte da regi?o VP1 do genoma dos EV, foram usados na t?cnica de rea??o em cadeia da polimerase precedida de transcri??o reversa (RT-PCR) para a detec??o molecular viral, produzindo um amplicon de 356 nucleot?deos. Entre os anos de 1996 a 2006, 538 amostras fecais de casos suspeitos de PFA, provenientes de toda a Regi?o Norte do Brasil, foram encaminhadas ao Instituto Evandro Chagas. Deste total, 78 (14,5%; 78/538) apresentaram efeito citop?tico nas duas linhagens celulares. Posteriormente, 51 (65,4%; 51/78) foram positivas por RT-PCR. Os casos positivos de EV foram detectados nos Estados do Amazonas (14/51; 27,4%), Amap? (13/51; 25,5%), Par? (11/51; 21,6%), Rond?nia (10/51; 19,6%) e Tocantins (3/51; 5,9%). A identifica??o por sequenciamento gen?mico foi poss?vel em 40 amostras. Echov?rus (20 casos, nove sorotipos) e coxsackiev?rus (dez casos, tr?s sorotipos) foram as esp?cies predominantes encontradas. Os resultados deste estudo corroboram outros realizados no Brasil e no mundo, e servem de base para o melhor entendimento da epidemiologia molecular dos EV circulantes na Regi?o Amaz?nica e a associa??o de seus diversos sorotipos com casos de doen?as neurol?gicas. Enteroviruses (EV) of human origin belong to the family Picornaviridae, they have 30 nm in diameter, genome of single-stranded RNA, with fecal-oral transmission and have been related to outbreaks of several diseases of the central nervous system. This study aimed to identify the presence of non-polio enteroviruses (NPEV) isolated from cases of acute flaccid paralysis (AFP). Cell lines Hep-2C and RD that presented cytopathic effect were selected for further molecular tests. The viral RNA was extracted using QIAamp Viral RNA mini kit. Primers 222 and 292, which increased part of the VP1 region of EV genome, were used in the reverse transcription polymerase chain reaction (RT-PCR) technique for molecular virus detection, producing an amplicon of 356 nucleotides. From 1996 to 2006, 538 fecal samples of AFP suspected cases from Northern Brazil were sent to the Instituto Evandro Chagas. Of this total, 78 (14.5%, 78/538) presented cytopathic effect in both cell lines. Thereafter, 51 (65.4%, 51/78) were positive by RT-PCR. EV positive cases were detected in the States of Amazonas (14/51, 27.4%), Amap? (13/51, 25.5%), Par? (11/51, 21.6%), Rond?nia (10/51, 19.6%) and Tocantins (3/51, 5.9%). Genomic sequencing identification was performed in 40 samples. Echovirus (20 cases, nine serotypes) and coxsackievirus (ten cases, three serotypes) were the predominant species found. The results of this study corroborate others conducted in Brazil and in the world, and serve as basis for better understanding about molecular epidemiology of EV circulating in the Amazon Region and its connection with several serotypes cases of neurologic diseases.
Published: 2014
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15. Varicella-zoster virus: identification of genotypes in cases of varicella and herpes zoster in the Municipalities of Ananindeua, Bel?m and Marituba, Par? State, Brazil

Author: Marcos Rogério Menezes da Costa, Igor Brasil Costa, Jane Haruko Lima Kaiano, Talita Antônia Furtado Monteiros, Darleise de Souza Oliveira, Alexandre da Costa Linhares, Francisco Lúzio de Paula Ramos, and Rita Catarina Medeiros Sousa
Subjects: Gen?tipos, Polimorfismo de Nucleot?deo ?nico, Varicela, Strategy and Management, Mechanical Engineering, Herpesvirus Humano 3, Metals and Alloys, Rea??o em Cadeia da Polimerase / m?todo, Regi?o Metropolitana de Bel?m (PA), Herpes Zoster, Industrial and Manufacturing Engineering
Abstract: Universidade Federal do Par?. N?cleo de Medicina Tropical. Bel?m, PA, Brasil / Minist?rio da Sa?de. Secretaria de Vigil?ncia em Sa?de. Instituto Evandro Chagas. Ananindeua, PA, Brasil. Minist?rio da Sa?de. Secretaria de Vigil?ncia em Sa?de. Instituto Evandro Chagas. Ananindeua, PA, Brasil. Minist?rio da Sa?de. Secretaria de Vigil?ncia em Sa?de. Instituto Evandro Chagas. Ananindeua, PA, Brasil. Minist?rio da Sa?de. Secretaria de Vigil?ncia em Sa?de. Instituto Evandro Chagas. Ananindeua, PA, Brasil. Universidade Federal do Par?. N?cleo de Medicina Tropical. Bel?m, PA, Brasil / Minist?rio da Sa?de. Secretaria de Vigil?ncia em Sa?de. Instituto Evandro Chagas. Ananindeua, PA, Brasil. Minist?rio da Sa?de. Secretaria de Vigil?ncia em Sa?de. Instituto Evandro Chagas. Ananindeua, PA, Brasil. Minist?rio da Sa?de. Secretaria de Vigil?ncia em Sa?de. Instituto Evandro Chagas. Ananindeua, PA, Brasil. Universidade Federal do Par?. N?cleo de Medicina Tropical. Bel?m, PA, Brasil / Minist?rio da Sa?de. Secretaria de Vigil?ncia em Sa?de. Instituto Evandro Chagas. Ananindeua, PA, Brasil. Este ? um estudo do tipo transversal, em que o crit?rio cl?nico para inclus?o das amostras foi a suspeita cl?nica e/ou aparecimento dos sintomas da varicela ou herpes-zoster para identifica??o dos gen?tipos do v?rus da varicela-zoster (VVZ), por meio das t?cnicas de rea??o em cadeia da polimerase (PCR) e sequenciamento, com posterior pesquisa de polimorfismo de nucleot?deo ?nico (SNP). De novembro de 2008 a mar?o de 2012, foram coletadas 93 amostras (85 casos de varicela e oito de herpes-zoster) de swab nasal, oral e de secre??o de ves?culas rompidas em v?rios s?tios anat?micos (49 de indiv?duos do sexo masculino e 44 do feminino). As estirpes Dumas e Oka parental foram inclu?das como refer?ncia padr?o dos gen?tipos europeu (classe 1 ou 3) e japon?s (classe 2), respectivamente. Estirpes com variabilidade no SNP foram classificadas dentro de um grupo de gen?tipo denominado mosaico (classe 5). As classes 1 ou 3 foram identificadas em 11 amostras do VVZ, nove casos de varicela e dois casos de herpes-zoster, enquanto que duas amostras apresentaram o gen?tipo de classe 5, ambos casos de varicela. O gen?tipo classe 1 ou 3 foi encontrado nos Munic?pios de Ananindeua, Bel?m e Marituba, Estado do Par?, Brasil, sendo que o gen?tipo classe 5 apenas em Ananindeua. A PCR foi sens?vel em 13 amostras de material direto de ves?culas, enquanto que em amostras de saliva n?o se obteve sucesso. This is a cross-sectional study in which the clinical criteria for inclusion of the samples was the clinical suspicion and/or onset of symptoms of chickenpox or shingles in order to genotype varicella-zoster virus (VZV) by the polymerase chain reaction (PCR) and sequencing techniques, with a search of single nucleotide polymorphisms (SNP) subsequently. From November 2008 to March 2012, 93 samples (85 cases of varicella and eight herpes zoster) were collected from oral, nasal swab and secretion of ruptured vesicles in several anatomical sites (49 male and 44 female). Dumas and Oka parental strains were included as European genotypes of the reference standard (class 1 or 3) and Japanese (class 2) genotypes, respectively. Strains with the SNP variability were classified into a group of genotype describe as Mosaic (class 5). Classes 1 or 3 were identified in 11 samples of VZV, nine cases of varicella and two cases of herpes zoster, while two samples showed the class genotype 5, both cases of varicella. The genotype class 1 or 3 was found in the Municipalities of Ananindeua, Bel?m and Marituba, Par? State, Brazil, and genotype class 5 only in Ananindeua. PCR was sensitive in just 13 samples of direct vesicles material, but in saliva samples there were not positive results.
Published: 2016

16. Phylogenetic analysis of probable Non-human genes of group A rotaviruses isolated from children with acute gastroenteritis in Belém, Brazil

Author: Joana D'Arc Pereira Mascarenhas, Yvone Benchimol Gabbay, Jane Haruko Lima Kaiano, José Paulo Gagliardi Leite, Darivaldo Luz Neri, Luana da Silva Soares, Régis Piloni Maestri, Sylvia de Fátima dos Santos Guerra, Darleise de Souza Oliveira, Alexandre da Costa Linhares, and Yasmin Nascimento Farias
Subjects: Rotavirus, Genes, Viral, Genotype, viruses, Reassortment, Viral Nonstructural Proteins, Biology, medicine.disease_cause, Genome, Rotavirus Infections, law.invention, Evolution, Molecular, Feces, law, Virology, medicine, Animals, Humans, Diarreia Infantil / virologia, Gene, Phylogeny, Polymerase chain reaction, Genetic diversity, Base Sequence, Phylogenetic tree, Reverse Transcriptase Polymerase Chain Reaction, Transmission (medicine), Viral Core Proteins, Infant, Newborn, Genetic Variation, Infant, virus diseases, Gastroenteritis, Infectious Diseases, Child, Preschool, Acute Disease, Gen?tica, RNA, Viral, Capsid Proteins, Infec??es por Rotavirus, Brazil, Reassortant Viruses
Abstract: National Council for Scientific and Technological Development (CNPq), Bras?lia, Federal District, Brazil. Grant Number: 4742742008?0 Minist?rio da Sa?de. Secretaria de Vigil?ncia em Sa?de. Instituto Evandro Chagas. Ananindeua, PA, Brasil. Minist?rio da Sa?de. Secretaria de Vigil?ncia em Sa?de. Instituto Evandro Chagas. Ananindeua, PA, Brasil. Minist?rio da Sa?de. Secretaria de Vigil?ncia em Sa?de. Instituto Evandro Chagas. Ananindeua, PA, Brasil. Minist?rio da Sa?de. Secretaria de Vigil?ncia em Sa?de. Instituto Evandro Chagas. Ananindeua, PA, Brasil. Minist?rio da Sa?de. Secretaria de Vigil?ncia em Sa?de. Instituto Evandro Chagas. Ananindeua, PA, Brasil. Minist?rio da Sa?de. Secretaria de Vigil?ncia em Sa?de. Instituto Evandro Chagas. Ananindeua, PA, Brasil. Funda??o Oswaldo Cruz. Instituto Oswaldo Cruz. Department of Virology. Rio de Janeiro, RJ, Brazil. Minist?rio da Sa?de. Secretaria de Vigil?ncia em Sa?de. Instituto Evandro Chagas. Ananindeua, PA, Brasil. Funda??o Oswaldo Cruz. Instituto Oswaldo Cruz. Department of Virology. Rio de Janeiro, RJ, Brazil. Minist?rio da Sa?de. Secretaria de Vigil?ncia em Sa?de. Instituto Evandro Chagas. Ananindeua, PA, Brasil. Minist?rio da Sa?de. Secretaria de Vigil?ncia em Sa?de. Instituto Evandro Chagas. Ananindeua, PA, Brasil. Rotaviruses (RVs) are the main cause of acute viral gastroenteritis in both humans and young animals of various 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, and genome reassortment. The objective of this study was to characterize molecularly genes that encode structural and nonstructural proteins in unusual RV strains. The clinical specimens selected for this study were obtained from children and newborn 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 newborn with gastroenteritis were studied. Reassortment 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 to be 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.
Published: 2012
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17. Molecular characterization of human erythrovirus B19 strains obtained from patients with several clinical presentations in the Amazon region of Brazil

Author: Alexandre da Costa Linhares, Darleise de Souza Oliveira, Olinda Macêdo, Fernando L. Melo, Edison Luiz Durigon, Camila Malta Romano, Maria R. Freitas, Ronaldo Barros de Freitas, and Paolo Marinho de Andrade Zanotto
Subjects: Adult, Male, Lineage (genetic), Adolescent, Genotype, viruses, Parvovirus B19 Humano, Polymerase Chain Reaction, Virus, Parvoviridae Infections, Phylogenetics, Virology, Erythrovirus / isolamento & purifica??o, Parvovirus B19, Human, Humans, Child, Phylogeny, Aged, Aged, 80 and over, Parvoviridae, Erythrovirus, biology, Amazon rainforest, Infant, virus diseases, Bayes Theorem, Middle Aged, biology.organism_classification, Filogenia, Eritema Infeccioso, Infectious Diseases, Child, Preschool, DNA, Viral, biology.protein, Female, Antibody, Brazil
Abstract: Minist?rio da Sa?de. Secretaria de Vigil?ncia em Sa?de. Instituto Evandro Chagas. Bel?m, PA, Brasil. Universidade de S?o Paulo. Instituto de Ci?ncias Biom?dicas. Departamento de Microbiologia. Laborat?rio de Evolu??o Molecular e Bioinform?tica. S?o Paulo, SP, Brazil. Minist?rio da Sa?de. Secretaria de Vigil?ncia em Sa?de. Instituto Evandro Chagas. Bel?m, PA, Brasil. Universidade de S?o Paulo. Instituto de Ci?ncias Biom?dicas. Departamento de Microbiologia. Laborat?rio de Evolu??o Molecular e Bioinform?tica. S?o Paulo, SP, Brazil. Minist?rio da Sa?de. Secretaria de Vigil?ncia em Sa?de. Instituto Evandro Chagas. Bel?m, PA, Brasil. Minist?rio da Sa?de. Secretaria de Vigil?ncia em Sa?de. Instituto Evandro Chagas. Bel?m, PA, Brasil. Minist?rio da Sa?de. Secretaria de Vigil?ncia em Sa?de. Instituto Evandro Chagas. Bel?m, PA, Brasil. Universidade de S?o Paulo. Instituto de Ci?ncias Biom?dicas. Departamento de Microbiologia. Laborat?rio de Evolu??o Molecular e Bioinform?tica. S?o Paulo, SP, Brazil. Universidade de S?o Paulo. Instituto de Ci?ncias Biom?dicas. Departamento de Microbiologia. Laborat?rio de Virologia Cl?nica. S?o Paulo, SP, Brazil. Background: Human erythrovirus B19, endemic in the Amazon region since 1990, is associated with a wide spectrum of clinical presentations.Objectives: To assess the prevalence of erythrovirus B19 infection and the relative frequency of erythrovirus B19 genotypes in patients inthe Amazon region with various clinical presentations.Study design: A total of 487 clinical samples obtained from patients with symptoms suggestive of erythrovirus infection were tested usingspecific IgM and IgG antibody assays (ELISA) and PCR for viral DNA detection. Partial VP1 and VP2 regions were sequenced and genotyped by phylogenetic reconstruction.Results: B19 DNA was detected in 117 (24 por cento) of 487 samples. Of these, 106 (91 por cento) isolates were genotype 1 and 11 (9 por cento) were genotype 3.No genotype 2 was found. Genotype 1 had three clusters (A1, A2 and B) and all genotype 3 sequences were subtype 3b. All patients with hematological disorders within cluster B of genotype 1 were infected by the same B19 lineage, suggesting that this lineage of B19 may havebeen transmitted via transfusion of blood products.Conclusion: We reported two genotypes, 1 and 3b, with three genotype 1 clusters co-circulating in the Amazon region during the past 10 years.
Published: 2008
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18. Surto de norov?rus em um navio cruzeiro ao longo da costa brasileira, mar?o de 2011

Author: Alexandre da Costa Linhares, Yvone Benchimol Gabbay, Joana D'Arc Pereira Mascarenhas, Dielle Monteiro Teixeira, Luana Soares Barbagelata, Jones Anderson Monteiro Siqueira, Ian Carlos Gomes de Lima, Darleise de Souza Oliveira, Carla Gisele Ribeiro Garcia, Glicélia Cruz Aragão, and Daniele de Barros Galindo
Subjects: Gastroenterite / epidemiologia, Geography, Surtos de Doen?as, Vigil?ncia Epidemiol?gica / utiliza??o, Strategy and Management, Mechanical Engineering, Norovirus, Metals and Alloys, medicine, medicine.disease_cause, Humanities, Industrial and Manufacturing Engineering
Abstract: Minist?rio da Sa?de. Secretaria de Vigil?ncia em Sa?de. Instituto Evandro Chagas. Ananindeua, PA, Brasil. Minist?rio da Sa?de. Secretaria de Vigil?ncia em Sa?de. Instituto Evandro Chagas. Programa de P?s-Gradua??o em Virologia. Ananindeua, PA, Brasil. Minist?rio da Sa?de. Secretaria de Vigil?ncia em Sa?de. Instituto Evandro Chagas. Ananindeua, PA, Brasil. Minist?rio da Sa?de. Secretaria de Vigil?ncia em Sa?de. Instituto Evandro Chagas. Ananindeua, PA, Brasil. Universidade do Estado do Par?. Centro de Ci?ncias Biol?gicas e da Sa?de. Programa de P?s-Gradua??o em Biologia Parasit?ria na Amaz?nia. Bel?m, PA, Brasil. Minist?rio da Sa?de. Secretaria de Vigil?ncia em Sa?de. Instituto Evandro Chagas. Ananindeua, PA, Brasil. Minist?rio da Sa?de. Secretaria de Vigil?ncia em Sa?de. Instituto Evandro Chagas. Ananindeua, PA, Brasil. Vigil?ncia em Sa?de do Estado do Par?. Bel?m, PA, Brasil. Centro de Informa??es Estrat?gicas em Vigil?ncia em Sa?de. Bel?m, PA, Brasil. Minist?rio da Sa?de. Secretaria de Vigil?ncia em Sa?de. Instituto Evandro Chagas. Ananindeua, PA, Brasil. Minist?rio da Sa?de. Secretaria de Vigil?ncia em Sa?de. Instituto Evandro Chagas. Ananindeua, PA, Brasil. Outbreaks of gastrointestinal disease may occur on board of cruise ships due to the consumption of contaminated water or food, with a fast person-to-person transmission. Epidemiologic investigations carried out by Centers for Disease Control and Prevention in USA, have confirmed that more than 95% of gastroenteritis outbreaks in cruise ships are caused by norovirus (NoV). In March 2011 an outbreak of acute gastroenteritis (AGE) occurred on a cruise ship with 1,224 passengers and 554 crew members that sailed from Rio de Janeiro, Rio de Janeiro State (Southeast, Brazil) to Manaus, Amazonas State (North) and stopovers in Recife, Pernambuco State, and Fortaleza, Cear? State (Northeast) and Bel?m, Par? State (North). Epidemiological data were obtained and seven rectal swab samples were collected and tested for NoV detection and characterization by molecular techniques. A total of 53 persons (42 passengers and 11 crew members) developed AGE, 75.5% of whom were older than 60 years old. The symptoms duration was less than 48 h, most of the patients presenting vomiting (79.2%) and diarrhea (73.6%). Most of the cases varied from mild to moderate and only one patient needed parenteral rehydration. Cases of AGE were recorded in eight of 12 vessel floors, especially in the recreational areas. The seven rectal samples collected were all NoV-positive by RT-PCR and all NoV strains were genogrouped as GII by semi-nested PCR. The quantitative real-time PCR produced a 57.1% NoV positivity rate. The partial nucleotide sequencing classified five (71.4%) of these samples as GII.P4. Our findings highlight the need for continuous viral enteropathogens surveillance including cruise ships considering the increase of this kind of touristic option in Brazil. Os surtos da doen?a gastrointestinal podem ocorrer em navios cruzeiros devido ao consumo de ?gua e comida contaminadas, com r?pida transmiss?o de uma pessoa a outra. Investiga??es epidemiol?gicas realizadas pelos Centros de Controle e Preven??o de Doen?as, nos EUA, confirmaram que mais de 95% dos surtos de gastroenterite em navios s?o causados por norov?rus (NoV). Em mar?o de 2011, um surto de gastroenterite aguda (GEA) ocorreu em um cruzeiro com 1.224 passageiros e 554 tripulantes que partiu do Rio de Janeiro, Estado do Rio de Janeiro (Sudeste do Brasil) para Manaus, Estado do Amazonas (Norte) com paradas em Recife, Estado do Pernambuco, e Fortaleza, Estado do Cear? (Nordeste) e Bel?m, Estado do Par? (Norte). Dados epidemiol?gicos foram obtidos e sete amostras por swab retal foram coletadas e testadas para a detec??o e caracteriza??o do NoV por meio de t?cnicas moleculares. Um total de 53 pessoas (42 passageiros e 11 tripulantes) desenvolveram AGE, dos quais 75,5% eram maiores de 60 anos de idade. Os sintomas duraram menos de 48 h, a maioria dos pacientes apresentou v?mito (79,2%) e diarreia (73,6%). A maior parte dos casos obteve varia??o entre leve e moderado e apenas um paciente necessitou de reidrata??o parental. Casos de AGE foram registrados em oito dos 12 andares do navio, principalmente nas ?reas de recrea??o. As sete amostras de swab retal coletadas revelaram resultado positivo para NoV por meio de RT-PCR e todas as cepas foram genogrupadas como GII pelo semi-nested PCR. O PCR quantitativo em tempo real produziu uma taxa de positividade de 57,1% para NoV. O sequenciamento parcial de nucleot?deos classificou cinco (71,4%) das sete amostras como GII.P4. Os resultados destacam a necessidade de vigil?ncia cont?nua dos enteropat?genos virais, incluindo navios cruzeiros, considerando o aumento deste tipo de op??o tur?stica no Brasil.
Published: 2014

19. Genogroup I avian picobirnavirus detected in Brazilian broiler chickens: a molecular epidemiology study

Author: Darleise de Souza Oliveira, Balasubramanian Ganesh, Yvone Benchimol Gabbay, Joana D'Arc Pereira Mascarenhas, Rodrigo Vellasco Duarte Silvestre, Jane Haruko Lima Kaiano, Delana Andreza Melo Bezerra, and René Ribeiro da Silva
Subjects: Molecular Epidemiology, biology, Molecular epidemiology, Genotype, Broiler, Picobirnavirus, Genome, Viral, biology.organism_classification, Virology, Genome, DNA sequencing, Reverse transcription polymerase chain reaction, Feces, RNA Virus Infections, Animals, Gene, Chickens, Brazil, Phylogeny, Poultry Diseases
Abstract: Picobirnavirus (PBV) belongs to the family Picobirnaviridae. Picobirnaviruses contain a bisegmented dsRNA genome that is non-enveloped. A total of 85 pooled faecal samples were collected from the poultry of 37 farms from the Metropolitan Mesoregion of Belém (MMB), Pará state, Brazil. The viral RNA from each sample was analysed by PAGE and reverse transcriptase PCR (RT-PCR). For each county affected, at least one positive sample was selected, cloned and sequenced. The samples showed a positivity of 15.3 % (13/85) by PAGE and 49.4 % (42/85) by RT-PCR. Sequencing of these strains demonstrated a considerable RdRp gene heterogeneity that ranged from 56.1 to 100 % at the nucleotide level compared with prototypes of different species and water sewage, and from 50.3 to 100 % among themselves. Avian picobirnavirus (AvPBV) was detected in MMB broiler farms and showed a heterogeneous relationship with the prototypes used. This report includes what is believed to be the first gene sequencing of AvPBV in Brazilian broiler chickens.
Published: 2013

20. Genetic diversity of porcine sapoviruses in pigs from the Amazon region of Brazil

Author: J. M Hernandez, Thayara Morais Portal, Yvone Benchimol Gabbay, Joana D'Arc Pereira Mascarenhas, Jane Cecília Silveira de Matos, Aline Fernandes Barry, Fabrício Aleixo Dias, Jones Anderson Monteiro Siqueira, Daniel Camargo Stangarlin, and Darleise de Souza Oliveira
Subjects: Swine Diseases, Genetic diversity, Amazon rainforest, Swine, Significant difference, General Medicine, Biology, Virology, Sapovirus, Gastroenteritis, Family Caliciviridae, GenBank, Animals, Statistical analysis, Brazil, Phylogeny, Caliciviridae Infections
Abstract: Sapoviruses (SaVs) belong to the family Caliciviridae and are related to gastroenteritis viruses of humans and animals. These agents have been reported from several countries of the world and represent an important cause of economic loss. The Amazon area has a high degree of diversity of animals and plants, is located in the Northern Region of Brazil and accounts for a large part of the Brazilian territory. In this study, stool samples were collected from pigs during the phase of nursing (less than 28 days of age) and post-weaning (29 to 56 days of age) from January 2008 to February 2009. A total of 169 specimens (108 nursing and 61 post-weaning pigs) were tested by reverse transcription polymerase chain reaction (RT-PCR) using the primers p289/p290 for the detection of caliciviruses (CVs), i.e., SaVs and noroviruses (NoVs). Positive sequences were analyzed using BioEdit software (v. 7.1.3.0) and compared with other sequences registered in the GenBank database. A positive frequency of 12.4 % (21/169) was observed, and all of the viruses found were identified as SaVs, with 15 belonging to genogroup GIII (71.4 %), three to GVII-1 (14.3 %) and three to GVIII-2 (14.3 %). No NoVs were detected. The frequency of SaV infections was significantly higher in nursing pigs (17.6 %-19/108) than in post-weaning pigs (3.3 %-2/61). Considering the consistency of the samples, 14.7 % of the samples were classified as diarrheic, but statistical analysis demonstrated that there was no significant difference compared to normal specimens (p = 0.5795). For the first time, we have demonstrated the circulation of SaVs in pigs from the Amazon.
Published: 2013

21. Norovirus Diversity in Diarrheic Children from an African-Descendant Settlement in Belém, Northern Brazil

Author: Jones Anderson Monteiro Siqueira, Alexandre da Costa Linhares, Yvone Benchimol Gabbay, Consuelo Silva de Oliveira, Glicélia Cruz Aragão, Eliete da Cunha Araújo, Tulio Machado Fumian, Maria Silvia Sousa de Lucena, Joana D'Arc Pereira Mascarenhas, Luana da Silva Soares, Juliana das Merces Hernandez, Jane Haruko Lima Kaiano, and Darleise de Souza Oliveira
Subjects: RNA viruses, Epidemiology, viruses, lcsh:Medicine, medicine.disease_cause, Pediatrics, law.invention, Feces, fluids and secretions, law, Viral classification, Emerging Viral Diseases, Genotype, Gastrointestinal Infections, lcsh:Science, Child, Polymerase chain reaction, Multidisciplinary, biology, virus diseases, Gastroenteritis, Diarrhea, Infectious Diseases, Medicine, Pediatric Gastroenterology, medicine.symptom, Brazil, Research Article, Black People, Gastroenterology and Hepatology, Microbiology, Infectious Disease Epidemiology, Sapovirus, Dogs, Virology, medicine, Animals, Humans, Biology, Genetic diversity, Population Biology, lcsh:R, Norovirus, Outbreak, Genetic Variation, biology.organism_classification, Viral Disease Diagnosis, Emerging Infectious Diseases, Co-Infections, lcsh:Q, Mamastrovirus
Abstract: Norovirus (NoV), sapovirus (SaV) and human astrovirus (HAstV) are viral pathogens that are associated with outbreaks and sporadic cases of gastroenteritis. However, little is known about the occurrence of these pathogens in relatively isolated communities, such as the remnants of African-descendant villages ("Quilombola"). The objective of this study was the frequency determination of these viruses in children under 10 years, with and without gastroenteritis, from a "Quilombola" Community, Northern Brazil. A total of 159 stool samples were obtained from April/2008 to July/2010 and tested by an enzyme immunoassay (EIA) and reverse transcription-polymerase chain reaction (RT-PCR) to detect NoV, SaV and HAstV, and further molecular characterization was performed. These viruses were detected only in the diarrheic group. NoV was the most frequent viral agent detected (19.7%-16/81), followed by SaV (2.5%-2/81) and HAstV (1.2%-1/81). Of the 16 NoV-positive samples, 14 were sequenced with primers targeting the B region of the polymerase (ORF1) and the D region of the capsid (ORF2). The results showed a broad genetic diversity of NoV, with 12 strains being classified as GII-4 (5-41.7%), GII-6 (3-25%), GII-7 (2-16.7%), GII-17 (1-8.3%) and GI-2 (1-8.3%), as based on the polymerase region; 12 samples were classified, based on the capsid region, as GII-4 (6-50%, being 3-2006b variant and 3-2010 variant), GII-6 (3-25%), GII-17 (2-16.7%) and GII-20 (1-8.3%). One NoV-strain showed dual genotype specificity, based on the polymerase and capsid region (GII-7/GII-20). This study provides, for the first time, epidemiological and molecular information on the circulation of NoV, SaV and HAstV in African-descendant communities in Northern Brazil and identifies NoV genotypes that were different from those detected previously in studies conducted in the urban area of Belém. It remains to be determined why a broader NoV diversity was observed in such a semi-isolated community.
Published: 2013

22. Norovirus infection in children admitted to hospital for acute gastroenteritis in Bel?m, Par?, Northern Brazil

Author: Yvone Benchimol Gabbay, Thaís Cristina Nascimento de Carvalho, Jones Anderson Monteiro Siqueira, Joana D'Arc Pereira Mascarenhas, Glicélia Cruz Aragão, Alexandre da Costa Linhares, Darleise de Souza Oliveira, Maria Cleonice Aguiar Justino, Mirleide Cordeiro dos Santos, and Maisa Silvia de Sousa
Subjects: Male, Genotype, Gastroenterite / parasitologia, Sa?de P?blica, MEDLINE, Enzyme-Linked Immunosorbent Assay, medicine.disease_cause, Sensitivity and Specificity, Feces, Virology, medicine, Prevalence, Humans, Caliciviridae Infections, business.industry, Reverse Transcriptase Polymerase Chain Reaction, Norovirus, Infant, Newborn, Infant, virus diseases, Acute gastroenteritis, Gastroenteritis, Hospitalization, Infectious Diseases, Child, Preschool, Hospitais P?blicos, Female, business, Diarreia Infantil, Brazil
Abstract: Coordination of Improvement of Higher Education Personnel (CAPES) Evandro Chagas Institute, SVS, MS Federal University of Par? State. Tropical Medicine Center. Postgraduate Program in Tropical Diseases. Bel?m, PA, Brazil. Minist?rio da Sa?de. Secretaria de Vigil?ncia em Sa?de. Instituto Evandro Chagas. Ananindeua, PA, Brasil. Federal University of Par? State. Institute of Health Sciences. Bel?m, PA, Brazil. University of the Par? State. Postgraduate Program in Parasite Biology in the Amazon, Center for Biological and Health Sciences. Bel?m, PA, Brazil. Minist?rio da Sa?de. Secretaria de Vigil?ncia em Sa?de. Instituto Evandro Chagas. Ananindeua, PA, Brasil. Minist?rio da Sa?de. Secretaria de Vigil?ncia em Sa?de. Instituto Evandro Chagas. Ananindeua, PA, Brasil. Federal University of Par? State. Tropical Medicine Center. Bel?m, PA, Brazil. Minist?rio da Sa?de. Secretaria de Vigil?ncia em Sa?de. Instituto Evandro Chagas. Ananindeua, PA, Brasil. Minist?rio da Sa?de. Secretaria de Vigil?ncia em Sa?de. Instituto Evandro Chagas. Ananindeua, PA, Brasil. Minist?rio da Sa?de. Secretaria de Vigil?ncia em Sa?de. Instituto Evandro Chagas. Ananindeua, PA, Brasil. Noroviruses are the leading cause of epidemic, non-bacterial outbreaks of acute gastroenteritis, and are also a major cause of sporadic acute gastroenteritis in infants. The aim of the present study was to identify norovirus infections in children not infected by rotavirus admitted to hospital for acute gastroenteritis in Bel?m. A total of 348 fecal specimens were obtained from children with diarrhea aged less than 5 years, all of whom had tested negative for rotavirus, between May 2008 and April 2010. Fecal samples were screened for norovirus antigen using enzyme-immunoassay (EIA). Specimens were subjected to reverse-transcription polymerase chain reaction (RT-PCR) using the primers Mon432/434-Mon431/433 for detection of the GI and GII norovirus strains, respectively. Based on both methods, the overall norovirus positivity rate was 36.5% (127/348). Of the 169 samples collected in the first year, 44.4% (n?=?75) tested positive for norovirus using both methods, 35.5% (n?=?60) by EIA and 40.8% (n?=?69) by RT-PCR. Using RT-PCR as a reference standard, a sensitivity of 78.3%, specificity of 94%, and agreement of 87.6% were recorded. Genome sequencing was obtained for 22 (31.9%) of the 69 positive samples, of which 90.9% (20/22) were genotype GII.4d and 9.1% (2/22) were genotype GII.b. Norovirus infection was most frequent in children under 2 years of age (41.5%-115/277). The peak incidence (62.1%) of norovirus-related acute gastroenteritis in these patients (not infected by rotavirus) was observed in February 2010. These findings emphasize the importance of norovirus as a cause of severe acute gastroenteritis among children in Bel?m, Par?, Northern Brazil.
Published: 2013

23. Molecular epidemiology of avian rotavirus in fecal samples of broiler chickens in Amazon Region, Brazil, from August 2008 to May 2011

Author: Maria Cristina Manno, Joana D'Arc Pereira Mascarenhas, Yvone Benchimol Gabbay, Sílvio Orlan de Castro Chaves, Fernanda do Espírito Santo Sagica, Artur Silva, Darleise de Souza Oliveira, Jane Haruko Lima Kaiano, Delana Andreza Melo Bezerra, René Ribeiro da Silva, and Amauri Alcindo Alfieri
Subjects: Galinhas / virologia, Aves Dom?sticas, Filogenia, Rotav?rus, Fezes / virologia, Strategy and Management, Mechanical Engineering, Metals and Alloys, Rea??o em Cadeia da Polimerase Via Transcriptase Reversa / m?todos, Doen?as das Aves / virologia, Industrial and Manufacturing Engineering
Abstract: This study was partially supported by a grant from the Science and Technology Secretariat of Par? State, contract n? 067/2008 (Sedect/Fapespa/PA), and Instituto Evandro Chagas/SVS/MS, Brazil. Delana Andreza Melo Bezerra and Jane Haruko Lima Kaiano received a Grant fellowship from the Brazilian National Council for the Development of Science and Technology (CNPq). Minist?rio da Agricultura, Pecu?ria e Abastecimento. Laborat?rio Nacional Agropecu?rio. Bel?m, PA, Brasil. Minist?rio da Sa?de. Secretaria de Vigil?ncia em Sa?de. Instituto Evandro Chagas. Ananindeua, PA, Brasil. Minist?rio da Sa?de. Secretaria de Vigil?ncia em Sa?de. Instituto Evandro Chagas. Ananindeua, PA, Brasil. Universidade Federal Rural da Amaz?nia. Instituto de Sa?de e Produ??o Animal. Bel?m, PA, Brasil. Minist?rio da Sa?de. Secretaria de Vigil?ncia em Sa?de. Instituto Evandro Chagas. Ananindeua, PA, Brasil. Minist?rio da Sa?de. Secretaria de Vigil?ncia em Sa?de. Instituto Evandro Chagas. Ananindeua, PA, Brasil. Minist?rio da Sa?de. Secretaria de Vigil?ncia em Sa?de. Instituto Evandro Chagas. Ananindeua, PA, Brasil. Minist?rio da Agricultura, Pecu?ria e Abastecimento. Laborat?rio Nacional Agropecu?rio. Bel?m, PA, Brasil. Universidade Federal do Par?. Instituto de Ci?ncias Biol?gicas. Bel?m, PA, Brasil. Universidade Estadual de Londrina. Departamento de Medicina Veterin?ria Preventiva. Londrina, PR, Brasil. Minist?rio da Sa?de. Secretaria de Vigil?ncia em Sa?de. Instituto Evandro Chagas. Ananindeua, PA, Brasil. Enteric viruses cause avian diseases that result in economic losses. Avian Rotavirus (AvRV) is the most important virus associated with enteritis in poultry. The main goals of this study were to determine the prevalence of AvRV using molecular tests in broiler chickens (Gallus gallus) from the metropolitan mesoregion of Bel?m (Par? State, Brazil), to provide epidemiological information, and to compare the rotaviruses detected in this study with reference to strains by phylogenetic analysis. Pooled fecal samples were collected from 37 poultry farms. The samples were tested for the NSP4 gene using reverse transcription polymerase chain reaction (RT-PCR). In total, 35 (41.2%) of the 85 fecal samples were positive for NSP4. There were 19 (51.4%) farms with at least one poultry house positive for AvRV. Considering the distribution of positive samples by age, the chicks aged 31-45 days comprised 18 (51.4%) of the 35 rotavirus-positive samples. Analyzing the data by density population, the houses with more than 9 birds/m2 had 25 (86.2%) positive samples, showing that higher infection rates occurred in higher density houses. To confirm the RT-PCR results and perform phylogenetic analysis, 20 positive samples were selected for sequencing. The rotaviruses detected in our study were clustered in a single group and had 93.5 to 100% sequence identity at the nucleotide level. The most affected age group included broiler chickens older than 15 days. Climatic conditions and high population densities favored the spread of AvRV and supported its uniform maintenance between seasons. As viroses ent?ricas causam doen?as em aves que resultam em perdas econ?micas. O rotav?rus avi?rio (AvRV) ? o v?rus mais importante associado ? enterite em aves dom?sticas. Os principais objetivos deste estudo foram determinar a preval?ncia do AvRV, usando testes moleculares em frangos de corte (Gallus gallus) da mesorregi?o metropolitana de Bel?m (Estado do Par?, Brasil), fornecer informa??es epidemiol?gicas e comparar os rotav?rus detectados com as cepas atrav?s da an?lise filogen?tica. Amostras fecais foram coletadas em 37 avi?rios e foram testadas para o gene NSP4 usando a rea??o em cadeia da polimerase via transcri??o reversa (RT-PCR). No total, das 85 amostras fecais, 35 (41,2%) mostram-se positivas para NSP4. Foram encontrados 19 (51,4%) avi?rios com pelo menos um galp?o positivo para AvRV. Considerando a distribui??o de amostras positivas por idade, os frangos de 31-45 dias continham 18 (51,4%) das 35 amostras positivas para rotav?rus. Analisando os dados pela densidade populacional, os galp?es com mais de 9 aves/m? tiveram 25 (86,2%) amostras positivas, mostrando que ?ndices maiores de infec??o ocorreram em galp?es de maior densidade. Para confirmar os resultado da RT-PCR e realizar a an?lise filogen?tica, 20 amostras positivas foram selecionadas por sequ?ncia. Os rotav?rus detectados em nosso estudo foram reunidos em um ?nico grupo e tiveram 93,5 a 100% de identidade sequencial no n?vel de nucleot?deos. O grupo et?rio mais atingido incluiu frangos de corte com mais de 15 dias. As condi??es clim?ticas e a alta densidade populacional favoreceram a dissemina??o do AvRV e sua manuten??o uniforme entre as esta??es.
Published: 2013

24. Identification of rotavirus G and P genotypes in nursing and weaned piglets in the metropolitan region of Bel?m, Par? State, Northern Brazil

Author: Yvone Benchimol Gabbay, Jane Cecília Silveira de Matos, Alexandre da Costa Linhares, Daniel Stangarlin de Camargo, Alessilva do Socorro Lima de Oliveira, Sylvia de Fátima dos Santos Guerra, Darleise de Souza Oliveira, Joana D'Arc Pereira Mascarenhas, and Luana da Silva Soares
Subjects: Veterinary medicine, Rotavirus G, Gen?tipo, Weaned piglets, Strategy and Management, Mechanical Engineering, Metals and Alloys, Rotavirus / isolamento & purifica??o, Imunocromatografia / utiliza??o, Biology, Humanities, Industrial and Manufacturing Engineering, Infec??es por Rotavirus / gen?tica
Abstract: Universidade Federal do Par?. N?cleo de Medicina Tropical. Bel?m, PA, Brasil. Universidade Federal do Par?. N?cleo de Medicina Tropical. Bel?m, PA, Brasil. Universidade Federal do Par?. N?cleo de Medicina Tropical. Bel?m, PA, Brasil. Minist?rio da Sa?de. Secretaria de Vigil?ncia em Sa?de. Instituto Evandro Chagas. Ananindeua, PA, Brasil. Universidade Federal do Par?. N?cleo de Medicina Tropical. Bel?m, PA, Brasil. Minist?rio da Sa?de. Secretaria de Vigil?ncia em Sa?de. Instituto Evandro Chagas. Ananindeua, PA, Brasil. Minist?rio da Sa?de. Secretaria de Vigil?ncia em Sa?de. Instituto Evandro Chagas. Ananindeua, PA, Brasil. Minist?rio da Sa?de. Secretaria de Vigil?ncia em Sa?de. Instituto Evandro Chagas. Ananindeua, PA, Brasil. Minist?rio da Sa?de. Secretaria de Vigil?ncia em Sa?de. Instituto Evandro Chagas. Ananindeua, PA, Brasil. Rotaviruses are an important agent of diarrhea in piglets, primarily during nursing and post-weaning with severity. The aim of this study was to identify rotavirus G and P genotypes obtained from piglets reared at five piggeries in Bel?m, Par? State, Northern Brazil. Fecal specimens were collected from nursing and weaned piglets. A total of 172 samples were tested, of which 17 (9.9%) were positive for group A rotaviruses all of them from piglets kept at nursing and then were sequenced for the of VP7 and VP4 genotyping. The consistency of positive fecal samples were 53% (9/17) diarrheic, 23.5% (4/17) pasty and 23.5% (4/17) normal. The most common G genotype was G3 representing 53% (9/17), followed by G5 genotype (17%, 3/17). The P genotype recorded was P[23] corresponding to 23.5% (4/17). This study showed that rotaviruses circulated in swine herds in the metropolitan region of Bel?m, in Northern region. G3P[23] combination was recorded for the first time in Brazil. Os rotav?rus s?o um importante agente de diarreia em leit?es, principalmente durante a amamenta??o e p?sdesmame com gravidade. O objetivo deste estudo foi identificar os gen?tipos G e P de rotav?rus obtidos de leit?es de cinco criadouros em Bel?m, Estado do Par?, Norte do Brasil. Esp?cimes fecais foram coletados de leit?es em fase de amamenta??o e desmamados. Foi testado um total de 172 amostras, das quais 17 (9,9%) foram positivas para rotav?rus do grupo A, a partir de leit?es amamentados e, em seguida, foram sequenciadas pela genotipagem do VP7 e VP4. A consist?ncia das amostras de fezes positivas foi de 53% (9/17) diarreico, 23,5% (4/17) pastosa e 23,5% (4/17) normal. O gen?tipo G mais comum foi o G3 com 53% (9/17), seguido pelo gen?tipo G5 (17%, 3/17). O gen?tipo P registrado foi de P[23] que corresponde a 23,5% (4/17). Este estudo mostrou a circula??o dos rotav?rus em rebanhos su?nos na regi?o metropolitana de Bel?m, na Regi?o Norte. A combina??o de G3P[23] foi registrada pela primeira vez no Brasil.
Published: 2012

25. Identification of lineage III of G12 rotavirus strains in diarrheic children in the Northern Region of Brazil between 2008 and 2010

Author: Alexandre da Costa Linhares, Darivaldo Luz Neri, Patrícia dos Santos Lobo, Luana da Silva Soares, Joana D'Arc Pereira Mascarenhas, Sylvia de Fátima dos Santos Guerra, Euzeni Maria de Fátima Costa de Menezes, Alessilva do Socorro Lima de Oliveira, Régis Piloni Maestri, and Darleise de Souza Oliveira
Subjects: Diarrhea, Male, Rotavirus, Diarr?ia Infantil, medicine.medical_specialty, Acute diarrhea, Lineage (genetic), Genotype, viruses, Molecular Sequence Data, Biology, medicine.disease_cause, Rotavirus Infections, Medical microbiology, Regi?o Norte (BR), Virology, Human rotavirus, medicine, Humans, Phylogeny, Phylogenetic tree, Infant, virus diseases, General Medicine, Gastroenterite / virologia, Child, Preschool, Female, Vp7 gene, Brazil
Abstract: Minist?rio da Sa?de. Secretaria de Vigil?ncia em Sa?de. Instituto Evandro Chagas. Ananindeua, PA, Brasil. Minist?rio da Sa?de. Secretaria de Vigil?ncia em Sa?de. Instituto Evandro Chagas. Ananindeua, PA, Brasil. Minist?rio da Sa?de. Secretaria de Vigil?ncia em Sa?de. Instituto Evandro Chagas. Ananindeua, PA, Brasil. Minist?rio da Sa?de. Secretaria de Vigil?ncia em Sa?de. Instituto Evandro Chagas. Ananindeua, PA, Brasil. Minist?rio da Sa?de. Secretaria de Vigil?ncia em Sa?de. Instituto Evandro Chagas. Ananindeua, PA, Brasil. Minist?rio da Sa?de. Secretaria de Vigil?ncia em Sa?de. Instituto Evandro Chagas. Ananindeua, PA, Brasil. Minist?rio da Sa?de. Secretaria de Vigil?ncia em Sa?de. Instituto Evandro Chagas. Ananindeua, PA, Brasil. Minist?rio da Sa?de. Secretaria de Vigil?ncia em Sa?de. Instituto Evandro Chagas. Ananindeua, PA, Brasil. Minist?rio da Sa?de. Secretaria de Vigil?ncia em Sa?de. Instituto Evandro Chagas. Ananindeua, PA, Brasil. Minist?rio da Sa?de. Secretaria de Vigil?ncia em Sa?de. Instituto Evandro Chagas. Ananindeua, PA, Brasil. This study reports on the surveillance forrotavirus genotypes and the identification of G12 human rotavirus in the Northern Region of Brazil. Rotavirus positive samples were collected from children years of age with acute diarrhea from January 2008 to October2010. G2P[4] was the most prevalent genotype, accounting for 45.6 per cent (126/303) of cases. Five rotavirus strains bearing G12P[6] genotype specificity were detected. Phylogeneticanalysis of the VP7 gene showed that G12 strains clustered into lineage III. This is the first detection of G12 strainsfrom lineage III in Latin America, broadening the current evidence for the worldwide emergence of this genotype.
Published: 2012

26. Detection of avian group D rotavirus using the polymerase chain reaction for the VP6 gene

Author: René Ribeiro da Silva, Alexandre da Costa Linhares, Delana Andreza Melo Bezerra, Joana D'Arc Pereira Mascarenhas, Rodrigo Vellasco Duarte Silvestre, Jane Haruko Lima Kaiano, Darleise de Souza Oliveira, and Yvone Benchimol Gabbay
Subjects: Rotavirus, Avian, Genes, Viral, viruses, RT-PCR, Picobirnavirus, Group D rotaviruses, medicine.disease_cause, Sensitivity and Specificity, law.invention, Feces, law, Virology, medicine, Escherichia coli, Animals, Diarreia, Cloning, Molecular, Gene, Polyacrylamide gel electrophoresis, Antigens, Viral, Polymerase chain reaction, Phylogeny, DNA Primers, biology, VP6 gene, Base Sequence, Reverse Transcriptase Polymerase Chain Reaction, virus diseases, Reproducibility of Results, biology.organism_classification, Molecular biology, Reverse transcription polymerase chain reaction, Real-time polymerase chain reaction, RNA, Viral, Capsid Proteins, Electrophoresis, Polyacrylamide Gel, Infec??es por Rotavirus, Rea??o em Cadeia da Polimerase Via Transcriptase Reversa, Aves, Chickens
Abstract: This study was partially supported by a grant from Par? State Secretary of Science and Technology, contract no. 067/2008 (SEDECT/FAPESPA/PA). Minist?rio da Sa?de. Secretaria de Vigil?ncia em Sa?de. Instituto Evandro Chagas. Ananindeua, PA, Brasil. Minist?rio da Agricultura. Laborat?rio Nacional Agropecu?rio. Bel?m, PA, Brazil. Minist?rio da Sa?de. Secretaria de Vigil?ncia em Sa?de. Instituto Evandro Chagas. Ananindeua, PA, Brasil. Minist?rio da Sa?de. Secretaria de Vigil?ncia em Sa?de. Instituto Evandro Chagas. Ananindeua, PA, Brasil. Minist?rio da Sa?de. Secretaria de Vigil?ncia em Sa?de. Instituto Evandro Chagas. Ananindeua, PA, Brasil. Minist?rio da Sa?de. Secretaria de Vigil?ncia em Sa?de. Instituto Evandro Chagas. Ananindeua, PA, Brasil. Minist?rio da Sa?de. Secretaria de Vigil?ncia em Sa?de. Instituto Evandro Chagas. Ananindeua, PA, Brasil. Minist?rio da Sa?de. Secretaria de Vigil?ncia em Sa?de. Instituto Evandro Chagas. Ananindeua, PA, Brasil. Group D rotaviruses (RVs-D) have been documented in birds and, while they may be common in these animals, few molecular studies are available for this specific group. In this study, specific primers for the gene that encodes for the RVs-D VP6 protein were designed and used in a reverse transcription polymerase chain reaction (RT-PCR). Thirty pools of samples were tested by polyacrylamide gel electrophoresis (PAGE) yielding a 30% (9/30) positivity. These pools were subjected subsequently to RT-PCR, with a 53% (16/30) positivity rate. The sensitivity of the PCR assay was demonstrated up to a dilution of 5 ? 10?4 ng/L (0.5 pg/L) of the cloned VP6 gene. The four samples were sequenced and showed 90.8?91.1% similarity with regards to the RVs-D VP6 gene. To assess for specificity our RT-PCR was applied to nine samples known to contain enteric viral agents other than group D rotaviruses including picobirnavirus, rotavirus group A, and reovirus with negative results. Overall, the data confirm the specificity of the primers used for detecting the RVs-D by RT-PCR, suggesting that this assay can be used for diagnostic purposes.
Published: 2011

27. Evaluation of third-generation RIDASCREEN enzyme immunoassay for the detection of norovirus antigens in stool samples of hospitalized children in Bel?m, Par?, Brazil

Author: Maria Silvia Sousa de Lucena, Joana D'Arc Pereira Mascarenhas, Yvone Benchimol Gabbay, Luana da Silva Soares, Jones Anderson Monteiro Siqueira, Ana Lucia Monteiro Wanzeller, Alexandre da Costa Linhares, and Darleise de Souza Oliveira
Subjects: Microbiology (medical), Microscopia Eletr?nica de Transmiss?o, Genotype, Infec??es por Caliciviridae / diagn?stico, Biology, Norovirus / isolamento & purifica??o, medicine.disease_cause, Sensitivity and Specificity, Immunoenzyme Techniques, Feces, Gen?tipo, Infec??es por Caliciviridae / virologia, Antigen, Microscopy, Electron, Transmission, medicine, Humans, V?rion / ultraestrutura, Ant?genos Virais / an?lise, Crian?a Hospitalizada, Antigens, Viral, Caliciviridae Infections, medicine.diagnostic_test, RNA Viral / gen?tica, Fezes / virologia, Reverse Transcriptase Polymerase Chain Reaction, Norovirus, Virion, Infant, Sensibilidade e Especificidade, General Medicine, Sequence Analysis, DNA, Virology, T?cnicas Imunoenzim?ticas / m?todos, Third generation, Gastroenteritis, Gastroenterite / virologia, Infectious Diseases, Immunoassay, Child, Preschool, Gastroenterite / diagn?stico, An?lise de Sequ?ncia de DNA, Rea??o em Cadeia da Polimerase Via Transcriptase Reversa / m?todos, RNA, Viral, Child, Hospitalized, Brazil
Abstract: Minist?rio da Sa?de. Secretaria de Vigil?ncia em Sa?de. Instituto Evandro Chagas. Ananindeua, PA, Brasil. Minist?rio da Sa?de. Secretaria de Vigil?ncia em Sa?de. Instituto Evandro Chagas. Ananindeua, PA, Brasil. Minist?rio da Sa?de. Secretaria de Vigil?ncia em Sa?de. Instituto Evandro Chagas. Ananindeua, PA, Brasil. Minist?rio da Sa?de. Secretaria de Vigil?ncia em Sa?de. Instituto Evandro Chagas. Ananindeua, PA, Brasil. Minist?rio da Sa?de. Secretaria de Vigil?ncia em Sa?de. Instituto Evandro Chagas. Ananindeua, PA, Brasil. Minist?rio da Sa?de. Secretaria de Vigil?ncia em Sa?de. Instituto Evandro Chagas. Ananindeua, PA, Brasil. Minist?rio da Sa?de. Secretaria de Vigil?ncia em Sa?de. Instituto Evandro Chagas. Ananindeua, PA, Brasil. Minist?rio da Sa?de. Secretaria de Vigil?ncia em Sa?de. Instituto Evandro Chagas. Ananindeua, PA, Brasil. Noroviruses (NoVs) are major agents of gastroenteritis outbreaks and hospitalization worldwide. This study evaluated the sensitivity and specificity of the commercially available third-generation RIDASCREEN? Norovirus Enzyme Immunoassay (EIA) kit in comparison to the reverse transcription-polymerase chain reaction (RT-PCR) to detect NoVs in hospitalized children with gastroenteritis. An agreement of 88% (81/92) was observed when comparing EIA with RT-PCR. A sensitivity of 92% and a specificity of 83.3% were demonstrated. Eleven samples were positive by 1 method only (4 RT-PCR/7 EIA). Fourteen samples were sequenced and all classified as NoV genogroup GII-4. The 7 positive only by EIA were also evaluated by electron microscopy, and in 3 (42.9%) samples viral particles with a suggestive morphology of NoVs were visualized. These same samples were tested by seminested-RT-PCR with a positivity of 85.7%. The results obtained in this study demonstrated a significant improvement in the sensitivity and specificity of this updated assay
Published: 2011

28. Molecular characterization of norovirus, sapovirus and astrovirus in children with acute gastroenteritis from Bel?m, Par?, Brazil

Author: Yvone Benchimol Gabbay, Darleise de Souza Oliveira, Consuelo Silva de Oliveira, Alexandre da Costa Linhares, Mirleide Cordeiro dos Santos, Glicélia Cruz Aragão, and Joana D'Arc Pereira Mascarenhas
Subjects: Gastroenterite / etiologia, business.industry, Strategy and Management, Mechanical Engineering, Norovirus, Metals and Alloys, Industrial and Manufacturing Engineering, Sapovirus, Dados de Sequ?ncia Molecular, Medicine, Infec??es por Astroviridae, Rea??o em Cadeia da Polimerase / m?todos, Infec??es por Astroviridae / diagn?stico, business, Mamastrovirus
Abstract: Minist?rio da Sa?de. Secretaria de Vigil?ncia em Sa?de. Instituto Evandro Chagas. Ananindeua, PA, Brasil. Minist?rio da Sa?de. Secretaria de Vigil?ncia em Sa?de. Instituto Evandro Chagas. Ananindeua, PA, Brasil. Minist?rio da Sa?de. Secretaria de Vigil?ncia em Sa?de. Instituto Evandro Chagas. Ananindeua, PA, Brasil. Minist?rio da Sa?de. Secretaria de Vigil?ncia em Sa?de. Instituto Evandro Chagas. Ananindeua, PA, Brasil. Minist?rio da Sa?de. Secretaria de Vigil?ncia em Sa?de. Instituto Evandro Chagas. Ananindeua, PA, Brasil. Minist?rio da Sa?de. Secretaria de Vigil?ncia em Sa?de. Instituto Evandro Chagas. Ananindeua, PA, Brasil. Minist?rio da Sa?de. Secretaria de Vigil?ncia em Sa?de. Instituto Evandro Chagas. Ananindeua, PA, Brasil. A import?ncia dos norov?rus (NoVs), sapov?rus (SaVs) e astrov?rus humanos (HAstVs) como causa de surtos de gastrenterites j? est? bem definida. Entretanto, poucos estudos t?m descrito casos espor?dicos de gastrenterite aguda causados por esses agentes. O objetivo deste estudo foi determinar o papel destes v?rus na etiologia das gastrenterites agudas em crian?as atendidas durante uma vigil?ncia intensiva realizada em hospitais e ambulat?rios de Bel?m, Brasil, de mar?o a setembro de 2003. Um total de 305 esp?cimes fecais de pacientes com gastrenterite grave foram coletados e testados por rea??o em cadeia da polimerase precedida de transcri??o reversa (RT-PCR), utilizando iniciadores espec?ficos Mon 269 e Mon 270 para os HAstVs; p289 e p290 para os caliciv?rus humanos (HuCVs); e Mon 431/433 e Mon 432/434 para os NoVs. Sequenciamento dos amplicons de HAstV, HuCVs e NoVs, obtidos por RT-PCR, foi realizado usando os mesmos iniciadores. Das 305 amostras testadas, 96 (31,5%) apresentaram resultados positivos, sendo que 51 diagnosticadas como HuCVs, 40 como HAstVs e cinco infec??es mistas. Das 56 (18,4%) amostras de HuCVs sequenciadas, 30 foram NoVs (9,8%) pertencentes aos genogrupos GI-4 e GII-4, e 15 (4,9%) SaVs dos grupos GI-1, GI-2 e GII-1. HAstVs foram detectados em 45 (14,7%) das amostras, incluindo os gen?tipos 1, 8 e 2. Esta pesquisa ressalta a import?ncia destas viroses como causa de gastrenterite aguda e demonstra a circula??o de diferentes gen?tipos durante o per?odo de estudo. Estes resultados refor?am a necessidade de se estabelecer uma vigil?ncia intensiva das gastrenterite causadas por estes v?rus, de forma a poder avaliar o impacto da doen?a e monitorar os gen?tipos circulantes. The importance of norovirus (NoVs), sapovirus (SaVs) and human astrovirus (HAstVs) as causes of gastroenteritis outbreaks is already well-defined, but a few studies have described sporadic cases of acute gastroenteritis caused by these viral entities. The aim of this study was to determine the role of these viruses in the etiology of acute gastroenteritis in children enrolled to participate in hospital ? and emergency department ? based intensive surveillance carried out in Bel?m, Brazil, from March to September 2003. A total of 305 stool specimens from patients with severe gastroenteritis were collected and screened by reverse transcription followed by polymerase chain reaction (RT-PCR), using the specific primers Mon 269 and Mon 270 for HAstVs, p289 and p290 for human calicivirus (HuCVs), and Mon 431/433 and Mon 432/434 for NoVs. Sequencing of RT-PCR HAstV, HuCV and NoV amplicons was carried out using the same primers. Of the 305 samples tested, 96 (31.5%) were positive, with 51 diagnosed as HuCVs, 40 as HAstVs and five as mixed infections. Of the 56 (18.4%) HuCVs sequenced, 30 were NoVs (9.8%) of genogroups GI-4 and GII-4, and 15 (4.9%) were SaVs of types GI-1, GI-2 and GII-1. HAstVs, including genotypes 1, 8 and 2, were detected in 45 (14.7%) samples. This study has highlighted the importance of these viruses as causes of acute gastroenteritis and established the circulation of different genotypes during the study period. These results reinforce the need for establishing an intensive surveillance for gastroenteritis caused by these viruses to assess the burden of disease and to monitor the circulation of genotypes.
Published: 2010

29. Echovirus 30 associated with cases of aseptic meningitis in state of Pará, Northern Brazil

Author: Olinda Macêdo, Maria José L Lima, Marquete B. Santana, Edna da Silveira, Ana Lucia Monteiro Wanzeller, Darleise de Souza Oliveira, Maria de Lourdes Contente Gomes, and Ceyla Maria Oeiras de Castro
Subjects: Adult, Male, Microbiology (medical), lcsh:Arctic medicine. Tropical medicine, Echovirus, Adolescent, Genotype, lcsh:RC955-962, Molecular Sequence Data, lcsh:QR1-502, viral meningitis, Echovirus Infections, Biology, medicine.disease_cause, Polymerase Chain Reaction, lcsh:Microbiology, medicine, Viral meningitis, Humans, Meningitis, Aseptic, Child, Phylogeny, Aged, Base Sequence, Molecular epidemiology, Reverse Transcriptase Polymerase Chain Reaction, enterovirus, Infant, Aseptic meningitis, Middle Aged, medicine.disease, Virology, Enterovirus B, Human, Child, Preschool, RNA, Viral, Enterovirus, Female, Nested polymerase chain reaction, Meningitis, Brazil
Abstract: Investigation of the aetiology of viral meningitis in Brazil is most often restricted to cases that occur in the Southern and Southeastern Regions; therefore, the purpose of this study is to describe the viral meningitis cases that occurred in state of Pará, Northern Brazil, from January 2005-December 2006. The detection of enterovirus (EV) in cerebrospinal fluid was performed using cell culture techniques, RT-PCR, nested PCR and nucleotide sequencing. The ages of the 91 patients ranged from < one year old to > 60 years old (median age 15.90 years). Fever (87.1%), headache (77.0%), vomiting (61.5%) and stiffness (61.5%) were the most frequent symptoms. Of 91 samples analyzed, 18 (19.8%) were positive for EV. Twelve were detected only by RT- PCR followed by nested PCR, whereas six were found by both cell culture and RT-PCR. From the last group, five were sequenced and classified as echovirus 30 (Echo 30). Phylogenetic analyses revealed that Echo 30 detected in Northern Brazil clustered within a unique group with a bootstrap value of 100% and could constitute a new subgroup (4c) according to the phylogenetic tree described by Oberste et al. (1999). This study described the first molecular characterization of Echo 30 in Brazil and this will certainly contribute to future molecular analyses involving strains detected in other regions of Brazil.
Published: 2009

30. Evidence for zoonotic transmission of group C rotaviruses among children in Belém, Brazil

Author: Joana D'Arc Pereira Mascarenhas, Yvone Benchimol Gabbay, Baoming Jiang, Roger I. Glass, Alexandre da Costa Linhares, Cláudia Maria Oliveira Simões, Alessandra Abel Borges, Darleise de Souza Oliveira, Yuhuan Wang, and Célia Regina Monte Barardi
Subjects: Rotavirus, Sequence analysis, Molecular Sequence Data, Reoviridae, Biology, medicine.disease_cause, Polymerase Chain Reaction, Virus, Rotavirus Infections, Feces, Virology, Sequence Homology, Nucleic Acid, Zoonoses, medicine, Animals, Humans, Longitudinal Studies, Antigens, Viral, Phylogeny, Southern blot, DNA Primers, Swine Diseases, Molecular Epidemiology, Molecular epidemiology, Infant, Newborn, Infant, Sequence Analysis, DNA, biology.organism_classification, Blotting, Southern, Infectious Diseases, Child, Preschool, Capsid Proteins, Nested polymerase chain reaction, Brazil
Abstract: The prevalence and potential zoonotic transmission of group C rotavirus (RVC) were examined by testing fecal samples collected from children during a longitudinal study that was carried out in the outskirts of Belem, Brazil, from December 1982 to March 1986. The study involved a group of 30 children who were followed from birth to 3 years. Of the 77 samples tested from 29 children, 5 (6.5%) were positive for human and 3 (4%) for porcine RVC by using nested PCR assay with primers specific for VP6 gene of human or porcine RVC and by Southern hybridization using a probe specific for VP6 gene of both human and porcine RVC. In addition, a total of 59 fecal specimens from the 30th child were tested, 1 (1.7%) and 14 (23.7%) were positive for human and porcine RVC, respectively. Partial nucleotide sequences of VP6 gene demonstrated that the six human strains detected in Brazil were homologous with other human RVC, and 14 of the 17 porcine RVC strains examined showed a complete homology among themselves but differed slightly from the porcine Cowden strain, suggesting that a single porcine RVC strain was circulating in Belem. This study is the first to provide evidence for transmission of RVC from swine to human. They also indicate that both human and porcine RVC were endemic in Belem.
Published: 2008

31. Molecular epidemiology of astrovirus type 1 in Belém, Brazil, as an agent of infantile gastroenteritis, over a period of 18 years (1982-2000): identification of two possible new lineages

Author: Yvone Benchimol Gabbay, Márcio Roberto Teixeira Nunes, Alexandre da Costa Linhares, Marcos Brian Heinemann, Joana D'Arc Pereira Mascarenhas, José Paulo Gagliardi Leite, Liliany S. Nakamura, and Darleise de Souza Oliveira
Subjects: Cancer Research, Genotype, Lineage (evolution), Molecular Sequence Data, Astrovirus, Virology, Astroviridae Infections, Humans, Phylogeny, Genetics, Whole genome sequencing, Genetic diversity, Molecular Epidemiology, Phylogenetic tree, biology, Molecular epidemiology, Base Sequence, Incidence (epidemiology), Infant, Newborn, Genetic Variation, Infant, biology.organism_classification, Gastroenteritis, Infectious Diseases, Child, Preschool, Diarrhea, Infantile, Brazil, Mamastrovirus
Abstract: Human astroviruses (HAstV) are worldwide recognized as important viral enteropathogens during childhood. This study aims to determine the incidence, genetic diversity and intertype variability of HAstV-1 in children less than 5 years of age enrolled in several studies conducted in Bel´ em/Parand S˜ ao Lu´ os/MaranhBrazil, from December 1982 to May 2000. Using EIA and RT-PCR, an overall positivity of 6.1% (155/2.534) was achieved, of these, 140 were positive by RT-PCR. The analysis of a 348 bp ORF2 fragment revealed that HAstV-1 was the predominant genotype (85/140, 60.7%) throughout the 18 years of study. Phylogenetic analysis was performed for 81 of these strains, and 76 (93.8%) were genetically classified as HAstV-1a. The remainder of strains (n = 5) were assigned to possible new lineages, 1e and 1f. Four of these five strains were detected in 1983 and 1984, and the lineage 1a circulated during 10 consecutive years (1990/2000). Genome sequence variation was found among the HAstV-1 strains involving all lineages, but only five nucleotide changes translated into aminoacid changes over this period, suggesting that HAstV-1 was very stable. The data obtained in this study should be useful for further studies at molecular level, including improvement of disease surveillance based on molecular diagnostic tools, and vaccine development. © 2007 Elsevier B.V. All rights reserved.
Published: 2007

32. Prevalence of human astrovirus genotypes associated with acute gastroenteritis among children in Belém, Brazil

Author: Yvone Benchimol Gabbay, Darleise de Souza Oliveira, José Paulo Gagliardi Leite, Consuelo Silva de Oliveira, Luciana Damascena da Silva, Alexandre da Costa Linhares, Liliany S. Nakamura, Elielma L. Cavalcante-Pepino, Joana D'Arc Pereira Mascarenhas, and Talita Antônia Furtado Monteiro
Subjects: medicine.medical_specialty, Infec??es por Astroviridae / epidemiologia, Vacinas contra Rotavirus, Astrovirus, Cohort Studies, Feces, Open Reading Frames, Species Specificity, Virology, Astroviridae Infections, Epidemiology, Genotype, medicine, Humans, Typing, Genotyping, Phylogeny, Varia??o Gen?tica, Diarreia / virologia, Molecular Epidemiology, biology, Molecular epidemiology, business.industry, Reverse Transcriptase Polymerase Chain Reaction, Incidence, Gastroenterite, virus diseases, Infant, biology.organism_classification, Rotavirus vaccine, Rea??o em Cadeia da Polimerase, Diarrhea, Infectious Diseases, Case-Control Studies, Child, Preschool, Acute Disease, Diarrhea, Infantile, Capsid Proteins, Reagent Kits, Diagnostic, Seasons, medicine.symptom, business, Brazil, Mamastrovirus
Abstract: This study received financial support from SECTAM/FUNTEC/PA, IEC/ SVS/MS, IOC/FIOCRUZ, and CNPq Minist?rio da Sa?de. Secretaria de Vigil?ncia em Sa?de. Instituto Evandro Chagas. Ananindeua, PA, Brasil. Minist?rio da Sa?de. Secretaria de Vigil?ncia em Sa?de. Instituto Evandro Chagas. Ananindeua, PA, Brasil. Minist?rio da Sa?de. Secretaria de Vigil?ncia em Sa?de. Instituto Evandro Chagas. Ananindeua, PA, Brasil. Minist?rio da Sa?de. Secretaria de Vigil?ncia em Sa?de. Instituto Evandro Chagas. Ananindeua, PA, Brasil. Minist?rio da Sa?de. Secretaria de Vigil?ncia em Sa?de. Instituto Evandro Chagas. Ananindeua, PA, Brasil. Minist?rio da Sa?de. Secretaria de Vigil?ncia em Sa?de. Instituto Evandro Chagas. Ananindeua, PA, Brasil. Minist?rio da Sa?de. Secretaria de Vigil?ncia em Sa?de. Instituto Evandro Chagas. Ananindeua, PA, Brasil. Minist?rio da Sa?de. Secretaria de Vigil?ncia em Sa?de. Instituto Evandro Chagas. Ananindeua, PA, Brasil. Minist?rio da Sa?de. Secretaria de Vigil?ncia em Sa?de. Instituto Evandro Chagas. Ananindeua, PA, Brasil. Funda??o Oswaldo Cruz. Instituto Oswaldo Cruz. Department of Virology. Rio de Janeiro, RJ, Brazil. Worldwide human astroviruses (HAstV) have increasingly been recognized as causative agents of viral gastroenteritis, mainly in infants and young children. The aim of this study was to assess the epidemiology and genotype diversity of HAstVs detected in children who participated in a trial in Belem, Brazil with the rhesus human reassortant rotavirus vaccine tetravalent (RRV-TV). From April/1990 to August/1992, 624 diarrheic stool samples were tested by enzyme immunoassay (EIA) for HAstV, with a positive rate of 4.0 per cent. Reverse transcription-polymerase chain reaction (RT-PCR) was done in 129 samples (25 positive and 104 with twice the optical density (OD) value of negative control by EIA) being 33 positive. The overall positivity yielded by both methods was 5.4 per cent (34/624). Genotyping of the 33 positive samples was done by type-specific RT-PCR and confirmed by sequence analysis. Phylogenetic analysis was performed using a 348-bp fragment of the ORF2 region of the capsid gene. HAstV-1 was the most prevalent, accounting for 45.5 per cent of the isolates, followed by HAstV-2 (27.3 per cent), HAstV-3 (12.1 per cent), HAstV-4 (12.1 per cent), and HAstV-6 (3.0 per cent). The monthly distribution showed that HAstV-1 was predominant in the first year of study (May/1990 to May/1991) with highest prevalence in January/1991. HAstV-2 was predominant from July to November/1991 and HAstV-4 from September to October/1990. At 24 months of age, 30.6 per cent of children had been infected by HAstV. The clinical symptoms registered during HAstV associated-diarrhea were usually mild. These data highlight the circulation of the different HAstV genotypes in Belem during the study period.
Published: 2007

33. The 'pressure pan' evolution of human erythrovirus B19 in the Amazon, Brazil

Author: Alexandre da Costa Linhares, Fernando L. Melo, Lílian Walshkeller, Paolo Marinho de Andrade Zanotto, Egma Mayta Huatuco, Camila Malta Romano, Edison Luiz Durigon, Maria R. Freitas, Maria Luisa Barbosa, Darleise de Souza Oliveira, Ronaldo Barros de Freitas, and Edward C. Holmes
Subjects: Nonsynonymous substitution, Genes, Viral, Natural selection, viruses, Molecular Sequence Data, Evolutionary change, Erythema Infectiosum, Biology, law.invention, Evolution, Molecular, Parvoviridae Infections, law, Virology, Parvovirus B19, Human, Humans, Selection, Genetic, Evolutionary dynamics, Phylogeny, Genetics, High prevalence, Models, Genetic, Amazon rainforest, virus diseases, Bayes Theorem, Human Erythrovirus B19, Transmission (mechanics), Evolutionary biology, DNA, Viral, Capsid Proteins, B19 phylogeny, Erythrovirus evolution, Brazil
Abstract: To understand the evolutionary dynamics of human parvovirus B19, we analyzed VP1 and VP2 gene sequences of B19 sampled from Belém (Amazon), the city of São Paulo, Brazil and globally. Our analysis revealed a strikingly different pattern of evolutionary change for those viral lineages introduced into Belém, which exhibited a higher rate of nonsynonymous substitutions compared to those viruses sampled from other locations. We propose that difference this is due to the high prevalence of B19 in Belém (up to 85%) compared to other locations (prevalences of approximately 50%), which imposes a more intense selection pressure. Hence, those B19 lineages introduced into Belém experienced an elevated rate of amino acid change, driven by positive selection, in order to generate serial re-infections in a small web of transmission, which can be thought of as an evolutionary “pressure pan”.
Published: 2007

34. Detection of a neonatal human rotavirus strain with VP4 and NSP4 genes of porcine origin

Author: José Paulo Gagliardi Leite, Yvone Benchimol Gabbay, Jackson C. Lima, Luana da Silva Soares, Rosa Helena P Gusmão, Alexandre da Costa Linhares, Marcos Bryan Heinemann, Joana D'Arc Pereira Mascarenhas, Irene Trigueiros Araújo, and Darleise de Souza Oliveira
Subjects: Microbiology (medical), Rotavirus, Rotavirus / gen?tica, Genotype, Swine, viruses, Molecular Sequence Data, Rotavirus / isolamento & purifica??o, Reoviridae, Biology, Viral Nonstructural Proteins, medicine.disease_cause, Microbiology, Virus, Rotavirus Infections, Similarity (network science), medicine, Animals, Humans, Allele, Gene, Phylogeny, Glycoproteins, Toxins, Biological, chemistry.chemical_classification, Strain (chemistry), Infant, Newborn, virus diseases, General Medicine, biology.organism_classification, Amino acid, chemistry, Su?nos, Capsid Proteins, Brazil
Abstract: IEC/SVS, COTEC/SECTAM/PA, IOC/FIOCRUZ and PIBIC/CNPq/Brazil for financial and administrative support. Minist?rio da Sa?de. Secretaria de Vigil?ncia em Sa?de. Instituto Evandro Chagas. Bel?m, PA, Brasil. Funda??o Oswaldo Cruz. Instituto Oswaldo Cruz. Departamento de Virologia. Rio de Janeiro,RJ, Brazil. Minist?rio da Sa?de. Secretaria de Vigil?ncia em Sa?de. Instituto Evandro Chagas. Bel?m, PA, Brasil. Funda??o Oswaldo Cruz. Instituto Oswaldo Cruz. Departamento de Virologia. Rio de Janeiro,RJ, Brazil. Minist?rio da Sa?de. Secretaria de Vigil?ncia em Sa?de. Instituto Evandro Chagas. Bel?m, PA, Brasil. Funda??o Oswaldo Cruz. Instituto Oswaldo Cruz. Departamento de Virologia. Rio de Janeiro,RJ, Brazil. Minist?rio da Sa?de. Secretaria de Vigil?ncia em Sa?de. Instituto Evandro Chagas. Bel?m, PA, Brasil. Universidade do Estado do Par?. Bel?m, PA, Brazil. Minist?rio da Sa?de. Secretaria de Vigil?ncia em Sa?de. Instituto Evandro Chagas. Bel?m, PA, Brasil. Minist?rio da Sa?de. Secretaria de Vigil?ncia em Sa?de. Instituto Evandro Chagas. Bel?m, PA, Brasil. A human rotavirus strain (NB-150) was detected in stool samples from a neonate hospitalized for mild/moderate community-acquired diarrhoea. This baby lived in the outskirts of Bel?m, Brazil, under poor sanitation conditions. The NB-150 strain displayed a typical long electrophoretic pattern with 11 gene segments. It had two VP7 alleles, G1 and G4, and belonged to VP6 subgroup II. Aclose relatedness with human rotaviruses was shown for VP7 alleles: G1 (96.9?100 per cent) similarity at the amino acid level) and G4 (97.1?100 per cent similarity at the amino acid level). As for VP6, 95.1?97.5 per cent similarity at the amino acid level was noted. VP8* and NSP4 genes showed a close relatedness with those of porcine rotavirus strains, as follows: VP8* (95.0 per cent similarity at the amino acid level) and NSP4 (93.7?96.0 per cent similarity at the amino acid level). This is believed to be the first report in Brazil of a rotavirus infection involving a strain with G1 and G4 alleles, with VP8* and NSP4 genes of porcine origin. These findings strongly suggest the occurrence of interspecies transmission.
Published: 2007

35. First detection of a human astrovirus type 8 in a child with diarrhea in Bel?m, Brazil: comparison with other strains worldwide and identification of possible three lineages

Author: Yvone Benchimol Gabbay, Joana D'Arc Pereira Mascarenhas, Olinda Macêdo, Marcos Bryan Heinemann, Alexandre da Costa Linhares, José Paulo Gagliardi Leite, Rosa Helena P Gusmão, Darleise de Souza Oliveira, and Lilian S Nakamura
Subjects: Microbiology (medical), lcsh:Arctic medicine. Tropical medicine, lcsh:RC955-962, viruses, Nucleotide sequencing, lcsh:QR1-502, Biology, Diarr?ia Infantil / virologia, Homology (biology), lcsh:Microbiology, Mamastrovirus / classifica??o, astrovirus type 8, molecular characterization, Feces, Open Reading Frames, Astroviridae Infections, medicine, Humans, Sequence variation, Belém, Reverse Transcriptase Polymerase Chain Reaction, Mamastrovirus / isolamento & purifica??o, Infant, virus diseases, Human astrovirus, Virology, Diarrhea, Astrovirus type 8, Diarrhea, Infantile, Female, medicine.symptom, Brazil, Mamastrovirus
Abstract: Minist?rio da Sa?de. Secretaria de Vigil?ncia em Sa?de. Instituto Evandro Chagas. Ananindeua, PA, Brasil. Minist?rio da Sa?de. Secretaria de Vigil?ncia em Sa?de. Instituto Evandro Chagas. Ananindeua, PA, Brasil. Minist?rio da Sa?de. Secretaria de Vigil?ncia em Sa?de. Instituto Evandro Chagas. Ananindeua, PA, Brasil. Minist?rio da Sa?de. Secretaria de Vigil?ncia em Sa?de. Instituto Evandro Chagas. Ananindeua, PA, Brasil. Minist?rio da Sa?de. Secretaria de Vigil?ncia em Sa?de. Instituto Evandro Chagas. Ananindeua, PA, Brasil. Universidade Estadual do Par?. Bel?m, PA, Brasil. Universidade Federal de Minas Gerais. Escola de Veterin?ria. Belo Horizonte, MG, Brasil. Minist?rio da Sa?de. Secretaria de Vigil?ncia em Sa?de. Instituto Evandro Chagas. Ananindeua, PA, Brasil. Funda??o Oswaldo Cruz. Instituto Oswaldo Cruz. Departmento de Virologia. Rio de Janeiro, RJ, Brasil. This study describes the genetic relationships of the first human astrovirus type-8 (HAstV-8) detected in Bel?m-Brazil, during a public hospital-based study. This strain was compared with other HAstV-8 strains identified elsewhere which have sequences available at GeneBank. The regions ORF1a (primers Mon348/Mon340) and ORF2 (primers Mon269/Mon270) were analyzed by nucleotide sequencing and a high similarity rate was observed among the Bel?m strain and other HAstV-8 strains. In ORF1a, homology values of 93-100% were detected, and in ORF2 96-99%. Considering the sequence variation (7%) observed in ORF2 region, it was suggested that HAstV-8 strains could be divided in three different lineages.
Published: 2007

36. Molecular characterization of VP4 and NSP4 genes from rotavirus strains infecting neonates and young children in Bel?m, Brazil

Author: Clarissa Silva Lima, Alexandre da Costa Linhares, Yvone Benchimol Gabbay, Darleise de Souza Oliveira, Olinda Macêdo, Joana D'Arc Pereira Mascarenhas, José Paulo Gagliardi Leite, Luana da Silva Soares, Sylvia de Fátima dos Santos Guerra, and Jackson C. Lima
Subjects: Rotavirus, Cancer Research, Genes, Viral, Genotype, Rotavirus / gen?tica, Swine, Molecular Sequence Data, Biology, Viral Nonstructural Proteins, medicine.disease_cause, Rotavirus Infections, Interspecies transmission, Data sequences, Virology, medicine, Animals, Humans, Amino Acid Sequence, Gene, Antigens, Viral, Rotavirus / classifica??o, Phylogeny, Glycoproteins, Toxins, Biological, Genetics, Sequence Homology, Amino Acid, Reverse Transcriptase Polymerase Chain Reaction, Infant, Newborn, Infant, RNA-Binding Proteins, Infectious Diseases, Close relationship, Child, Preschool, Capsid Proteins, Brazil, Infec??es por Rotavirus / gen?tica
Abstract: Minist?rio da Sa?de. Secretaria de Vigil?ncia em Sa?de. Instituto Evandro Chagas. Ananindeua, PA, Brasil. Minist?rio da Sa?de. Secretaria de Vigil?ncia em Sa?de. Instituto Evandro Chagas. Ananindeua, PA, Brasil. Minist?rio da Sa?de. Secretaria de Vigil?ncia em Sa?de. Instituto Evandro Chagas. Ananindeua, PA, Brasil. Minist?rio da Sa?de. Secretaria de Vigil?ncia em Sa?de. Instituto Evandro Chagas. Ananindeua, PA, Brasil. Minist?rio da Sa?de. Secretaria de Vigil?ncia em Sa?de. Instituto Evandro Chagas. Ananindeua, PA, Brasil. Minist?rio da Sa?de. Secretaria de Vigil?ncia em Sa?de. Instituto Evandro Chagas. Ananindeua, PA, Brasil. Minist?rio da Sa?de. Secretaria de Vigil?ncia em Sa?de. Instituto Evandro Chagas. Ananindeua, PA, Brasil. Universidade Federal de S?o Paulo. Departamento de Inform?tica em Sa?de. S?o Paulo, SP, Brazil. Minist?rio da Sa?de. Secretaria de Vigil?ncia em Sa?de. Instituto Evandro Chagas. Ananindeua, PA, Brasil. Funda??o Oswaldo Cruz. Instituto Oswaldo Cruz. Departamento de Virologia. Rio de Janeiro, RJ, Brazil. Several reports have identified P[6] specificities in humans and in animals in different countries of the world, but few sequence data are available in public databases. In this work we have characterized the VP4 strains bearing P[6] specificity and NSP4 genotypes among diarrheic young children and diarrheic and non-diarrheic neonates from three studies previously conducted in Bel?m, Northern region of Brazil. As the to VP8* fragment, we observed a close relationship to both human prototypes of lineage P[6]-Ia (bootstrap of 99 per cent) and porcine sublineages Ib and Ic (89.2?98.1 per cent aa similarity and mean of 95). With regards to the NSP4, the samples clustered into genotypes A and B. Of note, of the 27 P[6] strains analyzed in the present study and classified as genotype B, 8 (29.6 per cent) were more similar to porcine prototypes when VP8* and NSP4 genes are compared, and were recovered, one from a neonate and seven from diarrheic children. These preliminary findings reinforce that further investigations are needed to assess the relative frequencies of P[6] strains in our region, as well as to investigate the potential for interspecies transmission involving humans and animals particularly pigs.
Published: 2007

37. Characterization of an astrovirus genotype 2 strain causing an extensive outbreak of gastroenteritis among Maxakali Indians, Southeast Brazil

Author: Yvone Benchimol Gabbay, Joana D'Arc Pereira Mascarenhas, José Paulo Gagliardi Leite, Alexandre da Costa Linhares, Elielma L. Cavalcante-Pepino, Liliany S. Nakamura, Chequer Buffe Chamone, Simone Faria de Abreu, and Darleise de Souza Oliveira
Subjects: Veterinary medicine, medicine.medical_specialty, Reserva Maxakali (MG), Genotype, Population, Astrovirus, Disease Outbreaks, Feces, Mamastrovirus / gen?tica, Virology, Astroviridae Infections, Epidemiology, medicine, Humans, Minas Gerais (MG), education, Phylogeny, M?todos Epidemiol?gicos, education.field_of_study, biology, Reverse Transcriptase Polymerase Chain Reaction, Mamastrovirus / isolamento & purifica??o, Outbreak, virus diseases, Sapovirus, biology.organism_classification, Gastroenteritis, Diarrhea, Infectious Diseases, Surtos de Doen?as, Indians, North American, RNA, Viral, medicine.symptom, Brazil, Mamastrovirus
Abstract: This work received financial support from IEC-SVS, FUNTEC/PA, CNPq and FIOCRUZ Minist?rio da Sa?de. Secretaria de Vigil?ncia em Sa?de. Instituto Evandro Chagas. Bel?m, PA, Brasil. Funda??o Ezequiel Dias. Instituto Oct?vio Magalh?es. Divis?o de Epidemiologia e Controle de Doen?as. Belo Horizonte, MG, Brazil. Minist?rio da Sa?de. Secretaria de Vigil?ncia em Sa?de. Instituto Evandro Chagas. Bel?m, PA, Brasil. Minist?rio da Sa?de. Secretaria de Vigil?ncia em Sa?de. Instituto Evandro Chagas. Bel?m, PA, Brasil. Secretaria de Estado da Sa?de Minas Gerais. Coordena??o do Programa de Sa?de Ind?gena de Minas Gerais. Belo Horizonte, MG, Brazil. Minist?rio da Sa?de. Secretaria de Vigil?ncia em Sa?de. Instituto Evandro Chagas. Bel?m, PA, Brasil. Minist?rio da Sa?de. Secretaria de Vigil?ncia em Sa?de. Instituto Evandro Chagas. Bel?m, PA, Brasil. Funda??o Oswaldo Cruz. Instituto Oswaldo Cruz. Departamento de Virologia. Laborat?rio de Virologia Comparada. Rio de Janeiro, RJ, Brazil. Minist?rio da Sa?de. Secretaria de Vigil?ncia em Sa?de. Instituto Evandro Chagas. Bel?m, PA, Brasil. BACKGROUND: Human astroviruses (HAstVs) are a common cause of sporadic diarrhea in children but large outbreaks occur in children's day care centers, schools and in homes for the elderly. OBJECTIVES: To report a large outbreak of acute gastroenteritis that occurred in January 2004 among Indians from the Maxakali Reserve in the State of Minas Gerais, Brazil. STUDY DESIGN: Fecal samples from 46 Maxakali patients were collected and tested for a range of enteric pathogens. HAstVs was detected by EIA and strains were confirmed by RT-PCR and typed by direct sequencing. Clinical information was collected. RESULTS: The illness which affected more than 100 children under 6 years of age was characterized by watery diarrhea, fever and vomiting. HAstV-2 was the sole enteropathogen detected in 26 (56%) of 46 samples analyzed by EIA and/or RT-PCR. The 17 positive samples sequenced were identical in a 348bp region used for phylogenetic analysis and were classified as genotype 2. CONCLUSIONS: This is the first description of an outbreak of HAstV-2 gastroenteritis in a population of Brazilian Indians. The magnitude of the outbreak and the severity of the disease underscore the need to better understand the epidemiology of astrovirus and other enteric agents in these populations.
Published: 2006

38. SAPOVIRUSES IN CHILDREN WITH ACUTE GASTROENTERITIS FROM MANAUS , AMAZON REGION, BRAZIL, 2010-2011

Author: Darleise de Souza Oliveira, Yvone Benchimol Gabbay, Renato da Silva Bandeira, Maísa Silva de Sousa, Ian Carlos Gomes de Lima, Samya Thalita Picanço da Costa, Luciana Damascena da Silva, Tammy Kathlyn Amaral Reymão, Juliana das Merces Hernandez, Luana da Silva Soares, and Joana D'Arc Pereira Mascarenhas
Subjects: 0301 basic medicine, Veterinary medicine, lcsh:Arctic medicine. Tropical medicine, Genotype, lcsh:RC955-962, Biology, Sapovirus, Feces, 03 medical and health sciences, Infecções por SaVs, CIENCIAS DA SAUDE [CNPQ], Humans, Child, Genotyping, Amazon, Phylogeny, Caliciviridae Infections, Reverse Transcriptase Polymerase Chain Reaction, Amazon rainforest, Gastroenterite, General Medicine, Acute gastroenteritis, biology.organism_classification, Virology, Gastroenteritis, 030104 developmental biology, Infectious Diseases, Child, Preschool, Acute Disease, Original Article, Brazil
Abstract: Sapoviruses (SaVs) are responsible for acute gastroenteritis in humans, especially children and the elderly. In Brazil, data on SaVs infections are very limited, especially in Northern Brazil. Here, we investigated the occurrence of SaVs in samples from hospitalized children under ten years old that presented acute gastroenteritis. Positive samples were genotyped and phylogenetic analysis was performed using prototype strains sequences obtained from GenBank database. In total, 156 fecal samples were screened by RT-PCR for SaVs. A positivity rate of 3.8% (6/156) was found in children under three years of age. Four genotypes were detected: GI.I, GI.2 and GII.2?-GII.4?/GII.4, suggesting a possible inter-genotypes recombination. Most infections (83.3%) occurred between August and September. The positivity was similar to that found in other countries and genotyping demonstrated the presence of distinct genotypes. To our knowledge, this is the first study reporting the circulation of SaVs in Manaus, state of Amazonas, Amazon region, Brazil. HERNANDEZ, J.M.; SOUSA, M.S. Universidade Federal do Pará

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