RESUMO
It is suggested that bovine enteroviruses (BEV) are involved in the aetiology of enteric infections, respiratory disease, reproductive disorders and infertility. In this study, bovine faecal samples collected in different Brazilian states were subjected to RNA extraction, reverse transcription-polymerase chain reaction analysis and partial sequencing of the 5'-terminal portion of BEV. One hundred and three samples were tested with an overall positivity of 14.5%. Phylogenetic analysis clustered these BEV Brazilian samples into the Enterovirus F clade. Our results bring an important update of the virus presence in Brazil and contribute to a better understanding of the distribution and characterisation of BEV in cattle.
Assuntos
Doenças dos Bovinos/virologia , Infecções por Enterovirus/veterinária , Enterovirus Bovino/isolamento & purificação , Animais , Brasil/epidemiologia , Bovinos , Doenças dos Bovinos/epidemiologia , Infecções por Enterovirus/epidemiologia , Infecções por Enterovirus/virologia , Enterovirus Bovino/genética , FilogeniaRESUMO
Nebovirus is a new genus of viruses belonging to the Caliciviridae family recently characterized in cattle, and is associated with gastrointestinal disorders, such as diarrhoea, anorexia and intestinal lesions particularly in calves. The aim of this study was to investigate the prevalence of neboviruses in Brazilian cattle and analyse phylogenetically the virus strains detected. A prevalence of 4·8% of neboviruses in faecal samples from 62 head of cattle from different Brazilian states was detected. All positive animals were aged 96·0% nt (100% aa) sequence identity between the virus sequences in this study and >88·8% nt (>94·4% aa) identity with Newbury1/UK. Our results indicate, for the first time, the occurrence of neboviruses in Brazil as well as in South America, and the first Newbury1-like nebovirus found outside the UK.
Assuntos
Infecções por Caliciviridae/veterinária , Caliciviridae/classificação , Caliciviridae/isolamento & purificação , Doenças dos Bovinos/epidemiologia , Doenças dos Bovinos/virologia , Genótipo , Animais , Brasil/epidemiologia , Caliciviridae/genética , Infecções por Caliciviridae/epidemiologia , Infecções por Caliciviridae/virologia , Bovinos , Análise por Conglomerados , Diarreia/epidemiologia , Diarreia/veterinária , Diarreia/virologia , Fezes/virologia , Feminino , Gastroenterite/epidemiologia , Gastroenterite/veterinária , Gastroenterite/virologia , Humanos , Masculino , Filogenia , Prevalência , Homologia de Sequência do Ácido NucleicoRESUMO
The Infectious Bursal Disease Virus (IBDV) causes immunosuppression in young chickens. Advances in molecular virology and vaccines for IBDV have been achieved by viral reverse genetics (VRG). VRG for IBDV has undergone changes over time, however all strategies used to generate particles of IBDV involves multiple rounds of amplification and need of in vitro ligation and restriction sites. The aim of this research was to build the world's first VRG for IBDV by yeast-based homologous recombination; a more efficient, robust and simple process than cloning by in vitro ligation. The wild type IBDV (Wt-IBDV-Br) was isolated in Brazil and had its genome cloned in pJG-CMV-HDR vector by yeast-based homologous recombination. The clones were transfected into chicken embryo fibroblasts and the recovered virus (IC-IBDV-Br) showed genetic stability and similar phenotype to Wt-IBDV-Br, which were observed by nucleotide sequence, focus size/morphology and replication kinetics, respectively. Thus, IBDV reverse genetics by yeast-based homologous recombination provides tools to IBDV understanding and vaccines/viral vectors development.
Assuntos
Animais , Embrião de Galinha , Recombinação Homóloga , Vírus da Doença Infecciosa da Bursa/genética , Genética Reversa/métodos , Brasil , Células Cultivadas , Fibroblastos/virologia , Vetores Genéticos , Instabilidade Genômica , Vírus da Doença Infecciosa da Bursa/isolamento & purificação , Vírus da Doença Infecciosa da Bursa/fisiologia , Saccharomyces cerevisiae/genética , Transfecção , Cultura de Vírus , Replicação ViralRESUMO
The Infectious Bursal Disease Virus (IBDV) causes immunosuppression in young chickens. Advances in molecular virology and vaccines for IBDV have been achieved by viral reverse genetics (VRG). VRG for IBDV has undergone changes over time, however all strategies used to generate particles of IBDV involves multiple rounds of amplification and need of in vitro ligation and restriction sites. The aim of this research was to build the world's first VRG for IBDV by yeast-based homologous recombination; a more efficient, robust and simple process than cloning by in vitro ligation. The wild type IBDV (Wt-IBDV-Br) was isolated in Brazil and had its genome cloned in pJG-CMV-HDR vector by yeast-based homologous recombination. The clones were transfected into chicken embryo fibroblasts and the recovered virus (IC-IBDV-Br) showed genetic stability and similar phenotype to Wt-IBDV-Br, which were observed by nucleotide sequence, focus size/morphology and replication kinetics, respectively. Thus, IBDV reverse genetics by yeast-based homologous recombination provides tools to IBDV understanding and vaccines/viral vectors development.
Assuntos
Animais , Embrião de Galinha , Recombinação Homóloga , Vírus da Doença Infecciosa da Bursa/genética , Genética Reversa/métodos , Brasil , Células Cultivadas , Fibroblastos/virologia , Vetores Genéticos , Vírus da Doença Infecciosa da Bursa/isolamento & purificação , Saccharomyces cerevisiae/genética , Transfecção , Cultura de Vírus , Replicação ViralRESUMO
The Infectious Bursal Disease Virus (IBDV) causes immunosuppression in young chickens. Advances in molecular virology and vaccines for IBDV have been achieved by viral reverse genetics (VRG). VRG for IBDV has undergone changes over time, however all strategies used to generate particles of IBDV involves multiple rounds of amplification and need of in vitro ligation and restriction sites. The aim of this research was to build the world's first VRG for IBDV by yeast-based homologous recombination; a more efficient, robust and simple process than cloning by in vitro ligation. The wild type IBDV (Wt-IBDV-Br) was isolated in Brazil and had its genome cloned in pJG-CMV-HDR vector by yeast-based homologous recombination. The clones were transfected into chicken embryo fibroblasts and the recovered virus (IC-IBDV-Br) showed genetic stability and similar phenotype to Wt-IBDV-Br, which were observed by nucleotide sequence, focus size/morphology and replication kinetics, respectively. Thus, IBDV reverse genetics by yeast-based homologous recombination provides tools to IBDV understanding and vaccines/viral vectors development.