Rotaviruses are a major cause of diarrhea in humans and animals, including several mammalian and avian species. Using different PCR protocols, we report the occurrence of rotavirus A in 21 (53.84%; 21/39) from 39 fecal pool samples of broilers, layers, and broiler breeders from Brazilian avian farms. We typed the G5, G8, G11, G19, and P[31] genotypes.
Avian nephritis virus (ANV), which belongs to the family Astroviridae, is associated with different clinical manifestations (including enteric disorders). Despite being frequently found in the avian industry worldwide, information regarding genetic features of these viruses in Brazil is scarce. Therefore, sixty fecal sample pools (5-6 birds of the same flock), representing 60 poultry farms from six Brazilian States, were screened using an astrovirus-specific hemi-nested-PCR assay targeting the conserved ORF1b gene, followed by nucleotide sequencing of amplified products. PCR and phylogenetic analysis confirmed the detection of 21 positive samples to ANV (35 %). In order to investigate the genetic diversity represented by these viruses, amplification, cloning and phylogenetic analysis of the deduced amino acid sequence of ORF2 gene were attempted. Eight samples were successfully cloned (generating 32 clones in total) and sequenced. Based on phylogenetic analysis of ORF2, sequences defined in this study were classified into three genotypes: genotype 5, which has already been described in birds, and two other novel genotypes, tentatively named genotype 8 and 9, all of which occurred in single or mixed infections. Moreover, high intra-genotypic diversity and co-circulation of distinct strains in a same host population were observed. This study revealed the presence of new strains of ANV in Brazilian poultry and their circulation in commercial chicken flocks.
Rotaviruses are segmented double-stranded RNA viruses that cause gastroenteritis in mammals and birds. Here we describe the first partial nucleotide sequences of the structural protein VP6 from the genomes of group F rotaviruses that were detected in 5 out of 53 fecal samples (9.43%) from healthy broilers from Brazilian poultry farms based on reverse-transcriptase-PCR with primers designed for this study. The findings support the development of molecular detection systems, which can be used for the assessment of the distribution of rotavirus F in birds, their potential involvement in diseases, and their impact on poultry health.
Rotaviruses are responsible for the acute diarrhea in various mammalian and avian species. The nonstructural proteins NSP2 and NSP5 are involved in the rotavirus replication and the formation of viroplasm, cytoplasmic inclusion bodies within which new viral particles morphogenesis and viral RNA replication occur. There are few studies on the genetic diversity of those proteins; thus this study aims at characterizing the diversity of rotavirus based on NSP2 and NSP5 genes in rotaviruses circulating in Brazilian pig farms. For this purpose, 63 fecal samples from pig farms located in six different cities in the São Paulo State, Brazil, were screened by nested RT-PCR. Seven strains had the partial nucleotide sequencing for NSP2, whereas in six, the total sequencing for NSP5. All were characterized as genotype H1 and N1. The nucleotide identity of NSP2 genes ranged from 100% to 86.4% and the amino acid identity from 100% to 91.5%. For NSP5, the nucleotide identity was from 100% to 95.1% and the amino acid identity from 100% to 97.4%. It is concluded that the genotypes of the strains circulating in the region of study are in agreement with those reported in the literature for swine and that there is the possibility of interaction between human and animal rotaviruses.
RESUMO.-[Monitoramento e caracterização molecular de rotavírus do grupo D em criações comerciais brasileiras.] Rotavírus são agentes etiológicos de diarreia tanto em humanos como em várias espécies animais. Dados sobre rotavírus do grupo D (RVD) em aves são escassos, especialmente no Brasil. Nós detectamos RVD em 4 pools de conteúdo intestinal de frango de corte, poedeiras e matrizes de um total de 111 pools originários de 8 estados brasileiros, representando uma ocorrência de 3,6% a partir de uma RT-PCR específica para RVD, tendo como alvo o gene VP6. A árvore filogenética confirmou que as amostras brasileiras pertencem ao grupo D e três das sequências obtidas foram idênticas em termos de aminoácidos enquanto uma apresentou 99,5% de identidade com as demais. As sequências aqui definidas são semelhantes a outras sequências previamente definidas no Brasil, confirmando a natureza conservada da proteína VP6. INTRODUCTIONRotaviruses are members of the Reoviridae family, and they are a major cause of acute gastroenteritis in young children and several mammalian and avian species (Brüssow et al. 1992, Estes & Kapikian 2007. They are non-enveloped icosahedral particles and its genome comprises 11 segments of double-stranded RNA, which encodes six structural proteins (VP1 to VP4, VP6 and VP7) and six nonstructural proteins (NSP1 to NSP6). Based on antibody reactivity or genetic sequencing of structural protein VP6 of the internal capsid, rotaviruses have been classified into the groups A to G ( Brazil. E-mail: laila_andreia@hotmail.com Rotaviruses are etiological agents of diarrhea both in humans and in several animal species. Data on avian Group D rotaviruses (RVD) are scarce, especially in Brazil. We detected RVD in 4 pools of intestinal contents of broilers, layer and broiler breeders out of a total of 111 pools from 8 Brazilian states, representing an occurrence of 3.6%, by a specific RVD RT-PCR targeting the VP6 gene. Phylogenetic tree confirmed that the Brazilian strains belong to group D and 3 of the sequences were identical in terms of amino acid whereas one showed 99.5% identity with the others. The sequences described in this study are similar to other sequences previously detected in Brazil, confirming the conserved nature of the VP6 protein.
Rotaviruses are members of the family Reoviridae and are a common cause of acute diarrhea in many mammalian and avian species. They are non-enveloped icosahedral particles and their genome comprises 11 segments of double-stranded RNA, which encodes six structural proteins (VP1-4, VP6-7) and six nonstructural proteins (NSP1-6). Genotypes are defined based upon the diversity found in these genes and viral characterization plays a central role on epidemiological studies and prevention. Here we investigate the distribution of Brazilian RVAs genotypes in 8 chicken samples collected between 2008 and 2015 from different regions by RT-PCR, partial (Sanger) nucleotide sequencing and phylogenetic analysis from all rotavirus genes. Although the identified genotypes were typical from avian host species, when analyzed together, they form novel genetic constellations: G19-P[31]-I11-R6-C6-M7-A16-N6-T8-E10-H8 and G19-P[31]-I4-R4-C4-M4-A16-N4-T4-E4-H4. This study highlights that avian rotaviruses are widespread among commercial farms in Brazil, and the co-circulation of at least two different genomic constellations indicates that may present a way bigger genetic variability, that can be increased by the possible transmission events from other birds, lack of specific preventive measures, as well as the different viral evolution mechanisms.
Astrovirus is a common cause of enteritis in humans and domestic animals. Here we report the detection of turkey astrovirus type 1 (TAstV-1) and chicken astrovirus (CAstV) in avian farms. Sixty fecal sample pools (five or six birds of the same flock), from chickens without apparent clinical symptoms of enteric disease from farms located in six Brazilian states, were screened by an ORF1b PCR, followed by nucleotide sequencing of amplified products and phylogenetic analysis. Six samples tested positive for TAstV-1 and two for CAstV. One positive sample of each detected virus (TAstV-1 and CAstV) had the complete ORF2 sequenced. Data for the ORF2 sequence indicate that Brazilian TAstV-1 was divergent from TAstV-1 (United States), previously described infecting turkeys, and Brazilian CAstV clustered together with the U.K. group, subgroup B-II, associated with enteritis and growth retardation in chicks. This study provides updated information about CAstV and the first report of detection of TAstV-1 in Brazilian chickens, supporting the diagnostic of enteritis and epidemiologic surveillance in poultry health.
INTRODUCTIONGroup A rotavirus (RVA) is one of the main causative agents of diarrhea in a variety of animal species, as reported in several countries (Estes & Kapikian 2007). RVA infection is common in swine farms, and it results in decreased growth performance, increased mortality and economic loss (Papp et al. 2013).RVA belongs to Family Reoviridae, Subfamily Sedoreovirinae, Genus Rotavirus, and its genome consists of 11 double-stranded RNA (dsRNA) segments, coding for 6 structural (VP1, VP2, VP3, VP4, VP6, VP7) and 6 non-structural proteins (NSP1 through 6) (King et al. 2012).Diagnosis of RVA infection plays a central role in establishing measures for control and prevention. The methods currently available, the polyacrylamide gel electrophoresis (PAGE) technique (Herring et al. 1982) allows the detection of all rotavirus strains regardless of the group to which The results also demonstrated high intra-and inter-assay reproducibility (coefficient of variation ≤1.42%); thus, this method proved to be a fast and sensitive approach for the diagnosis of RVA in pigs.
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