RESUMO
Orf virus (ORFV) is the etiological agent of contagious ecthyma, a disease widely spread in the world that occasionally causes zoonotic infections. This work is the first molecular characterization of ORFV in Uruguay, where we analyzed twenty-one sheep samples, eighteen of which were recovered from thirteen ORFV outbreaks that occurred during 2004 to 2011 as well as three strains from a national vaccine. Phylogenetic analysis and the derived amino acid sequences from the B2L gene suggest that the Uruguayan virus do not form a unique cluster, with most of them displaying similarities with worldwide ORFV isolates as well as our vaccine strains.
Assuntos
Ectima Contagioso/virologia , Variação Genética , Vírus do Orf/genética , Ovinos/virologia , Animais , DNA Viral , Surtos de Doenças/veterinária , Ectima Contagioso/epidemiologia , Cabras/virologia , Vírus do Orf/classificação , Vírus do Orf/isolamento & purificação , Filogenia , Uruguai/epidemiologia , Proteínas Virais/genéticaRESUMO
Resumen La infección por el virus orf, también conocida como ectima contagioso, es reconocida una zoonosis ocupacional. Se diagnostica por lesiones cutáneas que evolu cionan rápidamente desde máculas a pápulas, vesículas y pústulas. Se presenta el caso clínico de una estudiante de medicina veterinaria que había tenido contacto con caprinos, clínicamente sanos y sin lesiones aparentes, hacía 19 días. Presentó dos lesiones vesiculares que coalescieron hasta formar una lesión de mayor tamaño rodeada por un halo eritematoso. Las lesiones fueron compatibles con la presentación clásica de las producidas por el virus orf en humanos. Se confirmó la presencia del virus orf mediante una RPC anidada del tejido de biopsia. Es uno de los primeros casos confirmados mediante técnicas moleculares en seres humanos en Chile.
Infection with the orf virus, also known as contagious ecthyma, is recognized as an occupational zoonosis worldwide. It is diagnosed by cutaneous lesions that progress rapidly from macules to papules, vesicles and pustules. The clinical case of a student of veterinary medicine who had had contact with goats, clinically healthy and without apparent lesions, which occured 19 days ago, is reported. She presented two vesicular lesions that coalesced to form a larger lesion surrounded by an erythematous halo. The lesions were compatible with the classical presentation of those produced by the orf virus in humans. The presence of the orf virus was confirmed by a nested PCR from biopsy tissue. It is one of the first cases confirmed by molecular techniques in humans in Chile.
Assuntos
Humanos , Animais , Feminino , Adulto Jovem , Vírus do Orf/isolamento & purificação , Ectima Contagioso/patologia , Vírus do Orf/patogenicidade , Biópsia , DNA Viral , Cabras , Doenças das Cabras/virologia , Chile , Reação em Cadeia da Polimerase , Dermatopatias Virais/patologiaRESUMO
Infection with the orf virus, also known as contagious ecthyma, is recognized as an occupational zoonosis worldwide. It is diagnosed by cutaneous lesions that progress rapidly from macules to papules, vesicles and pustules. The clinical case of a student of veterinary medicine who had had contact with goats, clinically healthy and without apparent lesions, which occured 19 days ago, is reported. She presented two vesicular lesions that coalesced to form a larger lesion surrounded by an erythematous halo. The lesions were compatible with the classical presentation of those produced by the orf virus in humans. The presence of the orf virus was confirmed by a nested PCR from biopsy tissue. It is one of the first cases confirmed by molecular techniques in humans in Chile.
Assuntos
Ectima Contagioso/patologia , Vírus do Orf/isolamento & purificação , Animais , Biópsia , Chile , DNA Viral , Feminino , Doenças das Cabras/virologia , Cabras , Humanos , Vírus do Orf/patogenicidade , Reação em Cadeia da Polimerase , Dermatopatias Virais/patologia , Adulto JovemAssuntos
Dermoscopia/métodos , Ectima Contagioso/diagnóstico , Vírus do Orf/isolamento & purificação , Criação de Animais Domésticos , Animais , Biópsia por Agulha , Ectima Contagioso/patologia , Ectima Contagioso/terapia , Dedos , Humanos , Imuno-Histoquímica , Pessoa de Meia-Idade , Exposição Ocupacional/efeitos adversos , Doenças Raras , Ovinos , Zoonoses/diagnóstico , Zoonoses/terapiaRESUMO
Orf virus (ORFV) is the etiological agent of contagious ecthyma (CE), a pustular dermatitis of sheep and goats. Outbreaks of ORFV have been observed in all geographical regions of the world, including Argentina. The origin and identity of Argentinian ORFVs are unknown, and no comparative or phylogenetic studies of these viruses have been performed. In this study, we described the sequencing and analysis of five ORFV molecular markers: a partial B2L gene (ORF011), VIR (ORF020), an envelope mature protein (ORF109), vIL10 (ORF127), and GIF (ORF117) from two particular Argentinian outbreaks of CE.
Assuntos
DNA Viral/genética , Ectima Contagioso/virologia , Vírus do Orf/classificação , Vírus do Orf/isolamento & purificação , Animais , Argentina , DNA Viral/química , Cabras , Dados de Sequência Molecular , Vírus do Orf/genética , Análise de Sequência de DNA , OvinosRESUMO
We herein describe the partial nucleotide sequencing and phylogenetic analysis of the B2L gene of seventeen Brazilian orf viruses (ORFV). Seventeen viruses were recovered from outbreaks of contagious ecthyma in sheep and goats in four states in Southern and Northeast country, and three from commercial vaccines. Most analyzed viruses were associated with outbreaks of classical contagious ecthyma, with lip, nostrils and labial commissure involvement, yet udder/teat, feet, vulvar and disseminated lesions were also reported in some cases. Nucleotide sequence analysis revealed a high degree of B2L similarity among sheep sequences (>99%) regardless the geographic origin, and a remarkable high identity for the two goat isolates (>99.8%), with similarity dropping to below 99% when comparing viruses from the two species. A phylogenetic tree grouped most sheep and goat viruses on different branches. In addition, sequence alignment allowed the identification of up to six scattered nucleotide changes that were predominant and more consistent in goat isolates, including a number of sequences from other continents. Thus, in spite of the high nucleotide similarity, different degrees of similarity and discrete nucleotide changes in the B2L gene may help in grouping ORFV viruses according to host species.
Assuntos
Ectima Contagioso/virologia , Doenças das Cabras/virologia , Vírus do Orf/genética , Animais , Sequência de Bases , Brasil/epidemiologia , Surtos de Doenças/veterinária , Ectima Contagioso/epidemiologia , Doenças das Cabras/epidemiologia , Cabras , Dados de Sequência Molecular , Vírus do Orf/classificação , Vírus do Orf/isolamento & purificação , Filogenia , Alinhamento de Sequência , Análise de Sequência de DNA , Carneiro DomésticoRESUMO
Orf virus (ORFV), the prototype of the genus Parapoxvirus, is the aetiological agent of contagious ecthyma (CE), a pustular dermatitis that afflicts domestic and wild small ruminants. CE is one of the most widespread poxvirus diseases in the world, causing public health impacts. Outbreaks of ORFV have been observed in all geographical regions of Brazil, affecting ovine and caprine herds. The origins, epidemiology and identity of Brazilian ORFVs are unknown, and no comparative or phylogenetic studies of these viruses have been performed. In the present study, we revisited CE outbreaks which occurred until 32 years ago, and we assessed, genetically, five viral isolates. We performed the sequencing and analysis of the three ORFV molecular markers: B2L gene, virus interferon resistance gene (VIR) and the vascular endothelial growth factor gene. Nucleotide and amino acid analysis of the analysed genes demonstrated that Brazilian ORFVs do not form a unique cluster, and presented more similarity to other worldwide ORFV samples than with each other. These data raise the questions of whether there are different worldwide ORFVs circulating in Brazil, or if all the Brazilian ORFV samples are of the same virus taken at distinct time points.
Assuntos
Surtos de Doenças/veterinária , Ectima Contagioso/virologia , Doenças das Cabras/virologia , Vírus do Orf/genética , Animais , Brasil/epidemiologia , Ectima Contagioso/epidemiologia , Marcadores Genéticos/genética , Doenças das Cabras/epidemiologia , Cabras , Vírus do Orf/isolamento & purificação , Estudos Retrospectivos , OvinosRESUMO
Ectima contagioso (EC) ou orf é uma doença infecto-contagiosa da pele causada pelo vírus do ectima contagioso (ECV) ou ORFV, que acomete principalmente ovinos e caprinos, e ocasionalmente o homem. Protótipo do gênero Parapoxvírus da família Poxviridae, está amplamente disseminado em todo mundo, inclusive no Brasil, especialmente na Região Nordeste, onde a caprinovinocultura é amplamente praticada para a produção de pele, carne e leite. Em Pernambuco, a doença ocorre de forma endêmica e pode ser confundida com enfermidades vesiculares, como a Febre Aftosa (FA), havendo a necessidade de diferenciação, sobretudo como suporte às ações do Programa Nacional de Erradicação da Febre Aftosa (PNEFA). Uma grande limitação no controle da doença é a dificuldade de replicação do vírus em cultivo celular, visando sua manipulação biotecnológica para a produção de vacinas. O objetivo deste trabalho é isolar e avaliar o comportamento de amostras de ECV em cultura primária de células de córnea fetal caprina, além de estabelecer um diagnóstico molecular eficiente para amostras de campo e avaliar o perfil filogenético das amostras de campo estudadas. Amostras de crostas de 22 ovinos e de sete caprinos que apresentavam sinais clínicos de EC, originários dos Estados de Pernambuco, Bahia, Sergipe e Paraíba, foram inoculadas em monocamadas de células epiteliais de córnea de feto caprino, durante sete passagens consecutivas, a intervalos semanais. Para realização do diagnóstico molecular foi utilizado para amplificação de um fragmento de 235pb do gene do envelope B2L o par de primers, PPP-3 e PPP-4 denominados pan-parapoxvirus. Observou-se em todas as passagens, a partir de 24 horas pós-infecção, efeito citopático (ECP) caracterizado pelo arredondamento celular, fusão com formação de pequenos sincícios, vacuolização e corpúsculos de inclusão citoplasmática, com intensidade de 25% a 100% de desprendimento da camada celular, que variou de acordo com a amostra viral. Conclui-se que as culturas de células primárias de córnea fetal caprina mostraram-se altamente permissíveis à replicação do ECV e que as amostras virais isoladas mostraram-se adaptadas ao cultivo utilizado, com pequena variação entre as amostras virais, e que, a identificação viral através da PCR é um método aplicável ao diagnóstico diferencial de enfermidades vesiculares, sobretudo como suporte às ações do PNEFA, no qual caprinos e ovinos são considerados animais sentinelas. A análise filogenética mostrou que as seqüências de ORFV do Nordeste do Brasil estão altamente relacionadas entre si, com seqüências idênticas acima de 99%. Além de apresentarem mais de 80% de identidade com seqüências de outros isolados de ORFV em todo mundo.
Contagious ecthyma is a severe and proliferative viral disease affecting ovine and caprine species, and eventually men, caused by the contagious ecthyma virus (ECV) of the Parapoxvirus genus. The disease is spread worldwide, including in Brazil, where in the state of Pernambuco, due to its endemicity, can be mistaken by other vesicular diseases such as the Foot and mouth Disease, which requires its diagnostic differentiation. Meanwhile, the control of the infection in endemic regions is limited due to the difficulties in replicating the virus in cell cultures, for the development of vaccines. The purpose of this work is to evaluate the behavior of ECV samples in primary cultures of fetal caprine cornea cells, and also to establish an efficient molecular diagnostic method for field samples, and to analyze the phylogenetic profile of the virus samples studied. Crust samples from twenty-two sheep and seven goats from the states of Pernambuco, Bahia, Sergipe and Paraiba presenting the clinical symptoms of EC were inoculated in cell monolayers, during seven consecutive passages at weekly intervals. During all passages, starting at 24 hours post-infection was observed a cytopathic effect (CE) characterized by cell rounding, cell fusion with the formation of small syncytia, cytoplasmic inclusion and vacuolization. The intensity of cell detachment from the layers ranged from 25% to 100% which varied according to the viral sample. We concluded that the primary cultures of the fetal caprine cornea cell appeared highly permissible to replication of ECV and the isolated samples of ECV seemed to adapt to the utilized culture with a slight variation among samples, and the identification by PCR showed to be an applicable method to the differential diagnosis of vesicular diseases, especially to support the actions of PNEFA, where goats and sheep are considered sentinel animals. The phylogenetic analysis showed that ORFV sequences from the northeastern of Brazil are highly correlated showing identical sequences up to 99%. In addition, they present more than 80% sequence identity with other isolates of ORFV worldwide.
Assuntos
Animais , Doenças das Cabras/diagnóstico , Técnicas e Procedimentos Diagnósticos/veterinária , Vírus do Orf/isolamento & purificação , Filogenia , Reação em Cadeia da Polimerase/veterináriaRESUMO
Orf virus is the etiological agent of contagious ecthyma, a severe exanthematic disease that affects small ruminants. Orf virus is zoonosis that is associated with occupational contact with infected animals in human disease. Clinically, contagious ecthyma is characterized by the appearance of vesicles, pustules, ulcers, and papillomatous proliferative lesions on the skin of the lips and nostrils. Here we describe a case of lethal cutaneous multifocal Orf virus infection in goats in the Amazon region of Brazil. Exanthematic lesions were collected and epidemiological and clinical data were obtained. Orf virus was detected using PCR amplification of the whole B2L, VIR, and VEGF open reading frame. Phylogenetic analysis revealed that this virus clustered together with the Orf virus samples isolated during classical contagious ecthyma. The present work is the first to report a severe proliferative Orf virus case in South America.
Assuntos
Doenças das Cabras/epidemiologia , Cabras/virologia , Vírus do Orf/isolamento & purificação , Vírus do Orf/patogenicidade , Dermatopatias Infecciosas/veterinária , Sequência de Aminoácidos , Animais , Brasil/epidemiologia , Ectima Contagioso/epidemiologia , Ectima Contagioso/patologia , Ectima Contagioso/virologia , Genes Virais , Doenças das Cabras/patologia , Doenças das Cabras/virologia , Doenças Labiais/epidemiologia , Doenças Labiais/patologia , Doenças Labiais/veterinária , Doenças Labiais/virologia , Dados de Sequência Molecular , Vírus do Orf/classificação , Vírus do Orf/genética , Filogenia , Alinhamento de Sequência , Dermatopatias Infecciosas/epidemiologia , Dermatopatias Infecciosas/virologiaRESUMO
BACKGROUND: Orf virus (ORFV), the prototype of the genus Parapoxvirus (PPV), is the etiological agent of contagious ecthyma, a severe exanthematic dermatitis that afflicts domestic and wild small ruminants. Although South American ORFV outbreaks have occurred and diagnosed there are no South American PPV major membrane glycoprotein B2L gene nucleotide sequences available. CASE PRESENTATION: an outbreak of ovine contagious ecthyma in Midwest Brazil was investigated. The diagnosis was based on clinical examinations and molecular biology techniques. The molecular characterization of the virus was done using PCR amplification, cloning and DNA sequencing of the B2L gene. The phylogenetic analysis demonstrated a high degree of identity with ORFV strains, and the isolate was closest to the ORFV-India 82/04 isolate. Another Brazilian ORFV isolate, NE1, was sequenced for comparative analysis and also showed a high degree of identity with an Asian ORFV strain. CONCLUSION: Distinct ORFV strains are circulating in Brazil. This is the first report on the phylogenetic analysis of an ORFV in South America.