RESUMO
BACKGROUND Serological evidence of West Nile virus (WNV) infection has been reported in different regions of Brazil from equine and human hosts but the virus had never been isolated in the country. OBJECTIVES We sought to identify the viral etiology of equine encephalitis in Espírito Santo state. METHODS We performed viral culture in C6/36 cells, molecular detection of WNV genome, histopathology and immunohistochemistry from horse cerebral tissue. We also carried out sequencing, phylogenetic analysis and molecular clock. FINDINGS Histopathologic analysis from horse cerebral tissue showed injury related to encephalitis and WNV infection was confirmed by immunohistochemistry. The virus was detected by reverse transcription quantitative polymerase chain reaction (RT-qPCR) from brain tissue and subsequently isolated in C6/36 cells. WNV full-length genome was sequenced showing the isolated strain belongs to lineage 1a. The molecular clock indicated that Brazilian WNV strain share the same common ancestor that were circulating in US during 2002-2005. MAIN CONCLUSIONS Here we report the first isolation of WNV in Brazil from a horse with neurologic disease, which was clustered into lineage 1a with others US WNV strains isolated in beginning of 2000's decade.
Assuntos
Humanos , Brasil/epidemiologia , Cavalos/anatomia & histologia , Vírus do Nilo Ocidental/patogenicidadeRESUMO
Abstract We report a case of encephalitis associated with Zika virus infection and reactivation of varicella-zoster virus in the central nervous system of a Brazilian child. This case raises the possibility that reactivation of the latent varicella-zoster virus may be a mechanism of neurological impairment induced by acquired Zika virus infection.
Assuntos
Humanos , Masculino , Criança , Herpesvirus Humano 3/isolamento & purificação , Encefalite por Varicela Zoster/complicações , Zika virus/isolamento & purificação , Infecção por Zika virus/complicações , Ativação Viral , DNA Viral/líquido cefalorraquidiano , Líquido Cefalorraquidiano/virologia , Encefalite por Varicela Zoster/diagnóstico , Eletroencefalografia , Infecção por Zika virus/diagnósticoRESUMO
This study showed that laboratory markers of recent infection by dengue, Zika or chikungunya arboviruses were detected in the biological samples of approximately one-third of patients with encephalitis, myelitis, encephalomyelitis or Guillain-Barré syndrome, in a surveillance programme in Piauí state, Brazil, between 2015-2016. Fever and myalgia had been associated with these cases. Since in non-tropical countries most infections or parainfectious diseases associated with the nervous system are attributed to herpesviruses, enteroviruses, and Campylobacter jejuni, the present findings indicate that in tropical countries, arboviruses may now play a more important role and reinforce the need for their surveillance and systematic investigation in the tropics.
Assuntos
Humanos , Vírus Chikungunya/genética , Vírus Chikungunya/imunologia , Vírus da Dengue/genética , Vírus da Dengue/imunologia , Zika virus/genética , Zika virus/imunologia , Doença Aguda , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Síndrome de Guillain-Barré/diagnóstico , Síndrome de Guillain-Barré/virologia , Encefalite/diagnóstico , Encefalite/virologia , Encefalomielite Aguda Disseminada/diagnóstico , Encefalomielite Aguda Disseminada/virologia , ELISPOT , Mielite Transversa/diagnóstico , Mielite Transversa/virologia , Doenças do Sistema Nervoso/diagnóstico , Doenças do Sistema Nervoso/virologiaRESUMO
Because an enriched environment (EE) enhances T-cell activity and T-lymphocytes contribute to immunopathogenesis during heterologous dengue virus (DENV) infections, we hypothesised that an EE increases dengue severity. To compare single serotype (SS) and antibody-enhanced disease (AED) infections regimens, serial intraperitoneal were performed with DENV3 (genotype III) infected brain homogenate or anti-DENV2 hyperimmune serum followed 24 h later by DENV3 (genotype III) infected brain homogenate. Compared AED for which significant differences were detected between the EE and impoverished environmental (IE) groups (Kaplan-Meyer log-rank test, p = 0.0025), no significant differences were detected between the SS experimental groups (Kaplan-Meyer log-rank test, p = 0.089). Survival curves from EE and IE animals infected with the AED regimen were extended after corticoid injection and this effect was greater in the EE than in the IE group (Kaplan-Meyer log-rank test, p = 0.0162). Under the AED regimen the EE group showed more intense clinical signs than the IE group. Dyspnoea, tremor, hunched posture, ruffled fur, immobility, pre-terminal paralysis, shock and death were associated with dominant T-lymphocytic hyperplasia and presence of viral antigens in the liver and lungs. We propose that the increased expansion of these memory T-cells and serotype cross-reactive antibodies facilitates the infection of these cells by DENV and that these events correlate with disease severity in an EE.
Assuntos
Animais , Feminino , Camundongos , Anticorpos Antivirais/imunologia , Anticorpos Facilitadores/imunologia , Vírus da Dengue/imunologia , Dengue/imunologia , Linfócitos B/imunologia , Dengue/virologia , Ecologia , Ativação Linfocitária/imunologia , Linfócitos T/imunologiaRESUMO
Dengue fever is the most important arbovirus infection found in tropical regions around the world. Dispersal of the vector and an increase in migratory flow between countries have led to large epidemics and severe clinical outcomes, such as dengue haemorrhagic fever and dengue shock syndrome. This study analysed the genetic variability of the dengue virus serotype 1 (DENV-1) in Brazil with regard to the full-length structural genes C/prM/M/E among 34 strains isolated during epidemics that occurred in the country between 1994-2011. Virus phylogeny and time of divergence were also evaluated with only the E gene of the strains isolated from 1994-2008. An analysis of amino acid differences between these strains and the French Guiana strain (FGA/89) revealed the presence of important nonsynonymous substitutions in the amino acid sequences, including residues E297 (Met→Thr) and E338 (Ser→Leu). A phylogenetic analysis of E proteins comparing the studied isolates and other strains selected from the GenBank database showed that the Brazilian DENV-1 strains since 1982 belonged to genotype V. This analysis also showed that different introductions of strains from the 1990s represented lineage replacement, with the identification of three lineages that cluster all isolates from the Americas. An analysis of the divergence time of DENV-1 indicated that the lineage circulating in Brazil emerged from an ancestral lineage that originated approximately 44.35 years ago.
Assuntos
Humanos , Vírus da Dengue/genética , RNA Viral/genética , Sequência de Aminoácidos , Brasil , Vírus da Dengue/classificação , Vírus da Dengue/isolamento & purificação , Genótipo , Dados de Sequência Molecular , FilogeniaRESUMO
Rocio virus (ROCV) is an encephalitic flavivirus endemic to Brazil. Experimental flavivirus infections have previously demonstrated a persistent infection and, in this study, we investigated the persistence of ROCV infection in golden hamsters (Mesocricetus auratus). The hamsters were infected intraperitoneally with 9.8 LD50/0.02 mL of ROCV and later anaesthetised and sacrificed at various time points over a 120-day period to collect of blood, urine and organ samples. The viral titres were quantified by real-time-polymerase chain reaction (qRT-PCR). The specimens were used to infect Vero cells and ROCV antigens in the cells were detected by immunefluorescence assay. The levels of antibodies were determined by the haemagglutination inhibition technique. A histopathological examination was performed on the tissues by staining with haematoxylin-eosin and detecting viral antigens by immunohistochemistry (IHC). ROCV induced a strong immune response and was pathogenic in hamsters through neuroinvasion. ROCV was recovered from Vero cells exposed to samples from the viscera, brain, blood, serum and urine and was detected by qRT-PCR in the brain, liver and blood for three months after infection. ROCV induced histopathological changes and the expression of viral antigens, which were detected by IHC in the liver, kidney, lung and brain up to four months after infection. These findings show that ROCV is pathogenic to golden hamsters and has the capacity to cause persistent infection in animals after intraperitoneal infection.
Assuntos
Animais , Cricetinae , Feminino , Anticorpos Antivirais/sangue , Infecções por Flavivirus/virologia , Flavivirus/imunologia , Viremia/virologia , Modelos Animais de Doenças , Técnica Indireta de Fluorescência para Anticorpo , Infecções por Flavivirus/imunologia , Infecções por Flavivirus/patologia , Imuno-Histoquímica , Mesocricetus , Reação em Cadeia da Polimerase em Tempo Real , RNA Viral/análiseRESUMO
The immunity of horses (n = 1401) against Saint Louis encephalitis virus (SLEV) was investigated in the Brazilian Amazon region (Bragança/Pará, Salvaterra/Pará, Macapá/Amapá and Rio Branco/Acre) and Maracaju, State of Mato Grosso do Sul, by the hemagglutination inhibition (HI) and plaque reduction neutralization (PRNT) tests. HI and neutralizing antibodies specific (monotypic reactivity, MR) for SLEV and other flaviviruses included in the tests were detected, as was cross-reactivity (CR) against flaviviruses. In the HI test, MR was observed in 248 (17.7 percent) serum samples, 137 of which were (55.2 percent) againstSLEV; CR was detected in 380 (27.1 percent). The frequency of MR against SLEV was significantly higher in Macapá and CR was significantly higher in Salvaterra. In the PRNT, neutralization of SLEV was observed in 713 (50.9 percent) samples, and the prevalence of neutralizing antibodies was significantly higher in Macapá than in Salvaterra (p = 0.0083). This study adds new data regarding the immunity of horses against SLEV in Brazil, and it confirms the wide distribution of SLEV and the diversity of flaviviruses in the country, as well as the apparent absence of disease in SLEV-infected horses.
A imunidade de equinos (n = 1401) contra o vírus da encefalite Saint Louis (SLEV) foi investigada na Amazônia brasileira (Bragança/PA, Salvaterra/PA, Macapá/AP e Rio Branco/AC) e Maracaju, no Estado do Mato Grosso do Sul, por meio detestes de inibição da hemaglutinação (IH) e neutralização por redução de placas (PRNT). Foram detectados anticorpos IH e neutralizantes específicos (reações monotípicas RM) para SLEV e outros flavivírus incluídos nos testes, assim como reações cruzadas para flavivírus. Pelo teste de IH, RM foram observadas em 248 (17,7 por cento) amostras de soro, 137 (55,2 por cento) para SLEV, e RC para flavivírus foram detectadas em 380 (27,1 por cento). A frequência de RM para SLEV e de RC foi significativamente maior em Macapá e Salvaterra, respectivamente. Pelo PRNT, foi observada a neutralização do SLEV em 713 (50,9 por cento) amostras, e a prevalência de anticorpos neutralizantes foi significativamente maior em Macapá, em comparação com Salvaterra (p = 0,0083). Este estudo traz novos dados a respeito da imunidade de equinos contra SLEV no Brasil, e confirma a ampla distribuição de SLEV e a diversidade de flavivírus no País, bem como a aparente ausência de doenças em equinos infectados por SLEV.
Assuntos
Humanos , Animais , Encefalite por Arbovirus , Encefalite de St. Louis/epidemiologia , Cavalos , Testes Sorológicos/métodosRESUMO
The dengue virus (DENV1-4) causes dengue fever and dengue hemorrhagic fever/dengue shock syndrome (DHF/DSS) in tropical and subtropical areas. The aim of this study was to evaluate the circulating genotypes of DENV. This was accomplished by sequencing the PrM and E genes of Brazilian isolates of DENV2 and DENV3 that were obtained between 1991 and 2008 from various geographic regions. Phylogenetic analyses of DENV2 demonstrated that the genotype III (Southeast Asian/American), in spite of several nucleotide and amino acid changes, was the only one that circulated over the past 19 years. Since its introduction in 2000, the DENV3 isolates that have been analyzed have all grouped intogenotype III (Indian subcontinent) and there has been no evidence of DENV3 belonging to other genotypes in this study.
O vírus dengue (DENV1-4) causa a dengue clássica e a febre hemorrágica da dengue / síndrome de choque da dengue (FHD/SCD) em regiões tropicais e subtropicais. O objetivo deste estudo foi avaliar os genótipos circulantes de DENV2 e DENV3 obtidos em distintas regiões geográficas no período de 1991 a 2008. As análises filogenéticas de DENV2 demonstraram que o genótipo III (Sudeste da Ásia/América), apesar das diversas alterações nucleotídicas e de aminoácidos, foi o único a circular durante os últimos 19 anos. Desde a sua introdução no estudo, em 2000, todas as amostras isoladas de DENV3 analisadas foram agrupadas no genótipo III (subcontinente indiano). Não foram encontradas evidências de que o DENV3 pertença a outros genótipos investigados.
Assuntos
Masculino , Feminino , Humanos , Vírus da Dengue/isolamento & purificação , Flavivirus/isolamento & purificação , Dengue Grave/classificaçãoRESUMO
Casos de doença febril aguda ocorridos em Boa Vista, estado de Roraima, Brasil foram investigados em julho de 2005. Amostras de sangue (n igual a 142) foram obtidas de pacientes clinicamente suspeitos de febre do dengue (FD). A tentativa de isolamento viral foi realizada em células C6/36 para pacientes com menos de cinco dias de doença (n igual a 96). As cepas isoladas foram identificadas pelo teste de imunofluorescência indireta (IFI) utilizando anticorpos monoclonais, bem como pela técnica de RT-PCR. Amostras de soro foram testadas pelo método de inibição da hemaglutinação (IH) e confirmadas pelo IgM-ELISA. O vírus dengue 3 (VDEN-3) foi isolado e detectado po RT-PCR em 41 pacientes. A região E de sete cepas foi sequenciada, sendo as mesmas identificadas como pertencentes ao genótipo III. Pelo teste de IH, 98 amostras de soros foram positivas para anticorpos IH para o vírus da Dengue, sendo os resultados confirmados pela detecção de anticorpos IgM para dengue. Clinicamente, todos os pacientes mostraram quadro de FD, sendo todas as faixas etárias afetadas independentes do sexo. A epidemia de dengue ocorrida em Roraima foi causada pelo VDEN-3, genótipo III, genótipo este que circula nas Américas desde 1994.