Detection and genetic characterization of Mamastrovirus 5 from Brazilian dogs

Abstract Mamastrovirus 5 (MAstV5), belonging to the Astroviridae (AstV) family, previously known as canine astrovirus or astrovirus-like particles, has been reported in several countries to be associated with viral enteric disease in dogs since the 1980s. Astroviruses have been detected in fecal samples from a wide variety of mammals and birds that are associated with gastroenteritis and extra enteric manifestations. In the present study, RT-PCR was used to investigate the presence of MAstV5 in 269 dog fecal samples. MAstV5 was detected in 26% (71/269) of the samples. Interestingly, all MAstV5-positive samples derived from dogs displaying clinical signs suggestive of gastroenteritis, other enteric viruses were simultaneously detected (canine parvovirus, canine distemper virus, canine coronavirus, canine adenovirus and canine rotavirus). Based on genomic sequence analysis of MAstV5 a novel classification of the species into four genotypes, MAstV5a-MAstV5d, is proposed. Phylogenetic analyses based on the ORF2 amino acid sequences, samples described herein grouped into the putative genotype 'a' closed related with Chinese samples. Other studies are required to attempt the clinical and antigenic implications of these astrovirus genotypes in dogs.

The circulation of human astrovirus genotypes in the Central West Region of Brazil

Out of 1,588 faecal samples of children taken from three locations of the Central West Region of Brazil, 57 were positive for astroviruses (HAstVs) using reverse transcription-polymerase chain reaction (RT-PCR). They were genotyped by nested RT-PCR and/or genomic sequencing. HAstV-1 (42.8 percent), HAstV-2 (23.2 percent), HAstV-3 (3.6 percent), HAstV-4 (14.3 percent) and HAstVs -5, -6, -7 and -8 (1.8 percent each) were detected. In Goiânia and Campo Grande, HAstV-1 was the most frequently detected genotype while in Brasília (DF) it was HAstV-2. Shifts in the circulation of astrovirus genotypes were observed in DF and Campo Grande. All samples collected by rectal swabs were viral negative. The astrovirus genotypes were detected in all age groups and there was no correlation between genotype and age group.

Caracterización molecular de calicivirus aislados de brotes de gastroenteritis ocurridos en la Argentina durante los años 2005 y 2006 / Molecular characterization of calicivirus strains detected in outbreaks of gastroenteritis occurring in Argentina during 2005 and 2006

Con el objetivo de determinar la incidencia de calicivirus, rotavirus y astrovirus en brotes de gastroenteritis ocurridos en diversas regiones de la Argentina durante los años 2005 y 2006, se analizaron muestras de materia fecal provenientes de 7 brotes con resultado de coprocultivo negativo. Para el diagnóstico de rotavirus se utilizó un ELISA comercial, mientras que para el diagnóstico de calicivirus y astrovirus se utilizó el método de RT-PCR. De las 74 muestras analizadas, 20 fueron positivas para calicivirus, 17 para rotavirus y una para astrovirus. No se identificaron infecciones virales mixtas. En 5 muestras positivas para calicivirus se secuenció una región del gen de la polimerasa; 4 de ellas correspondieron al género Norovirus y una al género Sapovirus. El análisis filogenético de las muestras secuenciadas determinó la presencia de norovirus de los genogrupos GI y GII; dentro de este último, se identificaron los genotipos GII-4, GII-b y GII-17. El análisis de la muestra en la cual se identificó sapovirus reveló la presencia del genotipo GI-1. Este estudio representa una continuación del análisis epidemiológico molecular de calicivirus asociados a brotes de gastroenteritis iniciado en 2004 y constituye la primera comunicación de la circulación de norovirus del genotipo GII-17 en la Argentina.

In order to determine the incidence of calicivirus, rotavirus and astrovirus in outbreaks of gastroenteritis occurring in different regions of Argentina during 2005 and 2006, fecal samples from seven nonbacterial outbreaks were analyzed. A commercial ELISA was used for rotavirus detection, while RT-PCRs were used for calicivirus and astrovirus. Of the 74 samples analyzed, 20 were calicivirus positive, 17 were rotavirus positive and one was astrovirus positive. No mixed infections were detected. A partial region of the RdRp gene was sequenced in five calicivirus positive-samples; 4 of them belonged to Norovirus genus and one to Sapovirus genus. The phylogenetic analysis of norovirus-positive-samples revealed the presence of strains from genogroups GI and GII; genotypes GII- 4, GII-b and GII-17 were identified within the latter. Phylogenetic the sapovirus-positive-sample revealed the presence of genotype GI-1. This study represents a follow-up of the of molecular epidemiology analysis of calicivirus associated to gastroenteritis outbreaks that have been carried out by our group since 2004, and constitutes the first report of the circulation of genotype GII-17 in Argentina.

Molecular characterization of astrovirus in stool samples from children in São Paulo, Brazil

The purpose of this study was to characterize astrovirus in faecal samples collected from children with and without diarrhea in São Paulo, Brazil, grouped into two sets: EPM and HU. Detection and genotyping were carried out using reverse transcription nested polymerase chain reaction (RT-PCR) with specific primers directed towards the genome open reading frame 2 (ORF2). Results for EPM set showed that 66/234 (28.2 percent) were positive: 28/94 (29.7 percent) from children with acute diarrhea, 14/45 (31.1 percent) with persistent diarrhea, and 9/55 (16.3 percent) from control individuals. No data was available for 15/40 (37.5 percent) of samples. Mixed infections with other viruses were found in 33 samples. In the HU, 18/187 (9.6 percent) were positive: 12/158 (7.6 percent) from individuals with acute diarrhea and 6/29 (20.7 percent) from control children. Four samples were mixed with other viruses. Out of 66 astrovirus positive EPM samples, 18 (27.2 percent) were characterized as human astrovirus type-1 (HAstV-1), two (3.0 percent) as HAstV-2, two (3.0 percent) as HAstV-3, and three (4.5 percent) as HAstV-8. Among 18 astrovirus positive HU samples, one (5.5 percent) was characterized as HAstV-1, six (33.3 percent) as HAstV-2, and one (5.5 percent) as HAstV-8. Two HAstV-8 genotyped samples were further confirmed by nucleotide sequencing. Our results shows that astroviruses are circulating in a constant manner in the population, with multiple serotypes, in higher frequency than it was described for other Brazilian regions. For the first time in Sao Paulo, Brazil, it was shown that astroviruses play an important role in children gastroenteritis, as described for most locations where they were detected.

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

This study describes the genetic relationships of the first human astrovirus type-8 (HAstV-8) detected in Belém-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 Belém strain and other HAstV-8 strains. In ORF1a, homology values of 93-100 por cento were detected, and in ORF2 96-99 por cento. Considering the sequence variation (7 por cento) observed in ORF2 region, it was suggested that HAstV-8 strains could be divided in three different lineages.