RESUMO
In different countries especially developing countries, treatment of urban wastewater might be ineffective removal of pathogens such as group A rotavirus. The objective of this study is to evaluate the efficiency of rotavirus removal in two wastewater treatment plants (WWTPs) in Isfahan, Iran. To meet the study objectives, 96 sewage samples from influent (n = 48) and final effluents (n = 48) were collected by grab sampling. Two different concentration methods included pellet and two-phase used for concentrating sewage samples. The presence of rotavirus antigens in all concentrated sewage samples was screened by enzyme-linked immunosorbent method. To analyze the study samples, real-time PCR technique with SYBR Green I fluorescent dye and nested multiplex PCR for rotavirus genotyping were utilized respectively. Result indicated positive rotavirus percentage in two methods of ELISA and real-time PCR was equal to 61.45% (59 cases) and 44.79% (43 cases). In addition, analyzing seasonal distribution of rotavirus shows different distributions as below: in spring (18.64%), summer (20.33%), autumn (35.60%), and winter (25.42%). Finally, rotaviruses illustrate significantly higher frequency in cold seasons. G10 and G1 types are considered the most, among common genotypes which identified in 11 (25.58%) and 5 (11.62%), out of the 43 positive samples in WWTPs, followed by non-typeable genotypes (13.95%) and mix genotypes (11.62%); and different genotypes including G2, G3, G4, G8, G9, and G12 were equal to 2.33, 9.30, 9.30, 2.33, 7, and 7% in the WWTPs, respectively. Such high prevalence underlines the significance of environmental surveillance. Also, to eliminate potential pathogens especially enteric viruses from sewage, the improvement of treatment systems is essential.
Assuntos
Rotavirus/isolamento & purificação , Águas Residuárias/virologia , Monitoramento Ambiental , Técnicas de Genotipagem , Humanos , Irã (Geográfico) , RNA Viral/genética , Rotavirus/genética , Estações do AnoRESUMO
The species A rotaviruses (RVA) are important gastroenteric pathogens that infect humans and animals. RVA genotype G3P[9] has been described in human-animal reassortment events, and the complexity of its hosts motivates the genetic investigation of this strain. Therefore, the aim of this study is to analyse a G3P[9] sample that was detected in a child with acute gastroenteritis. The 1A3739 sample featured the constellation G3P[9]-I18-R3-C3-Mx-A19-N3-T3-E3-H6. The sequence for VP3 gene was not obtained. The phylogeny showed a closer relationship among genes VP7, VP1, NSP3, NSP4, and NSP5 with genes of animal origin, such as chiropter, alpaca, equine, and simian. In addition, the genes VP6 and NSP1 belong to the new genotypes I18 and A19, respectively. The emergence of strains such as these can interfere with the effectiveness of the RVA vaccine, and continuous monitoring is therefore important. Additional studies are needed to determine the evolutionary source and to identify a possible reservoir of RVA in nature.
Assuntos
Gastroenterite/virologia , Genótipo , Vírus Reordenados/genética , Infecções por Rotavirus/virologia , Rotavirus/genética , Pré-Escolar , Evolução Molecular , Feminino , Humanos , Filogenia , Recombinação Genética , Rotavirus/isolamento & purificação , Análise de Sequência de DNARESUMO
Group A human rotaviruses (HuRVA) are causative agents of acute gastroenteritis. Six viral structural proteins (VPs) and six nonstructural proteins (NSPs) are produced in RV-infected cells. NSP4 is a diarrhoea-inducing viral enterotoxin and NSP4 gene analysis revealed at least 15 (E1-E15) genotypes. This study analysed the NSP4 genetic diversity of HuRVA G2P[4] strains collected in the state of São Paulo (SP) from 1994 and 2006-2010 using reverse transcription-polymerase chain reaction, sequencing and phylogenetic analysis. Forty (97.6%) G2P[4] strains displayed genotype E2; one strain (2.4%) displayed genotype E1. These results are consistent with the proposed linkage between VP4/VP7 (G2P[4]) and the NSP4 (E2) genotype of HuRVA. NSP4 phylogenetic analysis showed distinct clusters, with grouping of most strains by their genotype and collection year, and most strains from SP were clustered together with strains from other Brazilian states. A deduced amino acid sequence alignment for E2 showed many variations in the C-terminal region, including the VP4-binding domain. Considering the ability of NSP4 to generate host immunity, monitoring NSP4 variations, along with those in the VP4 or VP7 protein, is important for evaluating the circulation and pathogenesis of RV. Finally, the presence of one G2P[4]E1 strain reinforces the idea that new genotype combinations emerge through reassortment and independent segregation.
Assuntos
Adulto , Criança , Humanos , Antígenos Virais/isolamento & purificação , Glicoproteínas/genética , RNA Viral/genética , Rotavirus/genética , Toxinas Biológicas/genética , Proteínas não Estruturais Virais/genética , Sequência de Aminoácidos , Sequência de Bases , Brasil , Fezes/virologia , Variação Genética , Genótipo , Ligação Genética/genética , Técnicas Imunoenzimáticas , Dados de Sequência Molecular , Filogenia , Reação em Cadeia da Polimerase Via Transcriptase Reversa , RNA Viral/isolamento & purificação , Rotavirus/classificação , Rotavirus/imunologia , Alinhamento de SequênciaRESUMO
Accounting for an estimated 600,000 deaths worldwide each year, rotaviruses are recognized as the most important etiologic agents causing severe acute gastroenteritis among children under the age of five years. In Brazil, until rotavirus vaccination was established in the public health system in 2006, acute gastroenteritis striking children under five years and caused by these viruses was clearly associated with 3.5 million episodes of diarrhea, 650,000 visits to outpatient health care facilities, 92,000 hospitalizations, and 850 deaths each year. After the introduction of the rotavirus vaccine in Brazil in March 2006, studies all over the country have been comparing rotavirus genotypes circulating in the recent pre- and postvaccination era. Most of these studies have reported a high prevalence of the G2P[4] genotype and also a decrease in rotavirus detection all over Brazil after the introduction of the vaccine. So far, these are preliminary studies, as a longer period of time is necessary to establish if this high prevalence of G2P[4] is due to selective pressure by the vaccine on the circulating viruses or to a normal genotype fluctuation, and if it will have any impact on vaccine efficacy in the future. This review describes results from the most recent studies addressing this issue and on rotavirus genotypic variability in Brazil.