Identification and molecular characterization of Orf virus infection in occupationally exposed women in South America.

Contagious Ecthyma (CE) is a severe exanthematous dermatitis caused by the Orf virus (ORFV) that mainly affects domestic small ruminants such as sheep and goats. It is a worldwide-distributed occupational zoonosis, particularly infecting those in close contact with animals or animal products such as shepherds, farmers and veterinarians, among others. In the present work, we report the first human CE case confirmed in Argentina. A phylogenetic analysis based on four gene sequences of the isolated strain responsible for the disease showed that this isolate grouped with other ORFV sequences that caused reported CE cases in sheep from the same Argentine province. We also sequenced a sample from a Chilean human case reported in 2017, whose phylogenetic analysis showed that it groups together with other Argentine isolates from locations close to the border with Chile.

Outbreak of contagious ecthyma caused by Orf virus (Parapoxvirus ovis) in a vaccinated sheep flock in Uruguay.

Orf virus (ORFV) causes contagious ecthyma (CE), a highly transmissible, zoonotic disease of small ruminants. CE most commonly affects lambs and unvaccinated sheep. This work reports epidemiologic, clinicopathologic, and virologic findings in a CE outbreak in a vaccinated sheep flock in Uruguay and failure to detect ORFV in a commercial vaccine.

Genetic diversity of Orf virus isolated from sheep in Uruguay.

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.

Virus orf en humanos, confirmación molecular de un caso clínico en Chile / Orf virus in human, confirmation in case report from Chile

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.

OH1 from Orf Virus: A New Tyrosine Phosphatase that Displays Distinct Structural Features and Triple Substrate Specificity.

Viral tyrosine phosphatases such as VH1 from Vaccinia and Variola virus are recognized as important effectors of host-pathogen interactions. While proteins sharing sequence to VH1 have been identified in other viruses, their structural and functional characterization is not known. In this work, we determined the crystal structure of the VH1 homolog in the Orf virus, herein named OH1. Similarly to Variola and Vaccinia VH1, the structure of OH1 shows a dimer with the typical dual-specificity phosphatase fold. In contrast to VH1, the OH1 dimer is covalently stabilized by a disulfide bond involving residue Cys15 in the N-terminal helix alpha-1 of both monomers, and Cys15 is a conserved residue within the Parapoxvirus genus. The in vitro functional characterization confirms that OH1 is a dual-specificity phosphatase and reveals its ability to dephosphorylate phosphatidylinositol 3,5-bisphosphate, a new activity potentially relevant in phosphoinositide recycling during virion maturation.

[Orf virus in human, confirmation in case report from Chile]. / Virus orf en humanos, confirmación molecular de un caso clínico 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.

Outbreak of contagious echtyma in sheep in Rio Grande do Sul, Brazil / Surto de ectima contagioso em ovinos no Rio Grande do Sul, Brasil

Background: Contagious ecthyma is a viral disease caused by a Parapoxvirus, which affects primarily sheep and goats. The disease has a worldwide distribution and is characterized by cutaneous pustules and crusts mainly in the muzzle and lips. Although the disease has a worldwide distribution, there are few reports in the literature of contagious ecthyma outbreaks in Brazil. Moreover, this is an important disease, as well as causing huge economic losses due to high morbidity rates, is also a zoonosis occupational character. This report describes the epidemiological, clinical, and anatomopathological aspects of an outbreak of contagious echtyma in sheep in the State of Rio Grande do Sul, Brazil. Case: Cases were observed on January and February of 2016. Seventeen out of 45 Texel sheep were affected including five 4-6-month-old lambs, three 7-12-month-old male sheep and nine 2-year-old ewes. Before the outbreak, a Texel ram was introduced in the herd as replacement. Clinically, affected sheep had pustules, ulcers, and crusts in the lips, labial commissures, muzzle, and nasal bridge. They also presented dyspnea, submandibular and facial subcutaneous edema. One of the affected sheep was euthanized due to the poor prognosis. At necropsy, the lesions observed clinically were confirmed. Histopathology of the skin in the lips and muzzles showed marked acanthosis of the epidermis, [...](AU)

Outbreak of contagious echtyma in sheep in Rio Grande do Sul, Brazil / Surto de ectima contagioso em ovinos no Rio Grande do Sul, Brasil

Background: Contagious ecthyma is a viral disease caused by a Parapoxvirus, which affects primarily sheep and goats. The disease has a worldwide distribution and is characterized by cutaneous pustules and crusts mainly in the muzzle and lips. Although the disease has a worldwide distribution, there are few reports in the literature of contagious ecthyma outbreaks in Brazil. Moreover, this is an important disease, as well as causing huge economic losses due to high morbidity rates, is also a zoonosis occupational character. This report describes the epidemiological, clinical, and anatomopathological aspects of an outbreak of contagious echtyma in sheep in the State of Rio Grande do Sul, Brazil. Case: Cases were observed on January and February of 2016. Seventeen out of 45 Texel sheep were affected including five 4-6-month-old lambs, three 7-12-month-old male sheep and nine 2-year-old ewes. Before the outbreak, a Texel ram was introduced in the herd as replacement. Clinically, affected sheep had pustules, ulcers, and crusts in the lips, labial commissures, muzzle, and nasal bridge. They also presented dyspnea, submandibular and facial subcutaneous edema. One of the affected sheep was euthanized due to the poor prognosis. At necropsy, the lesions observed clinically were confirmed. Histopathology of the skin in the lips and muzzles showed marked acanthosis of the epidermis, [...]

Identification and molecular characterization of Orf virus in Argentina.

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.

Pathogenesis in lambs and sequence analysis of putative virulence genes of Brazilian orf virus isolates.

The parapoxvirus orf virus (ORFV) is the agent of contagious ecthyma, an ubiquitous mucocutaneous disease of sheep and goats that may present variable clinical presentations. We herein studied the pathogenesis of ORFV infection in lambs and analyzed three putative virulence genes of four Brazilian ORFV isolates. Lambs inoculated in the labial commissures with each ORFV isolate (n=4, viral titer 10(5.6) TCID50/ml) developed classical orf lesions, characterized by a progressive course of erythema/macules, vesicles, pustules and proliferative scabs. Lesions lasted an average of 22.9 days (18-26) and virus shedding was detected for approximately 24.6 days (18-30). Two isolates (SV269/11 and SV820/10) produced more severe, long-lasting lesions resulting in highest clinical scores. Lambs inoculated with isolate SV581/11 developed lesions markedly milder (lower clinical scores [p<0.05]) and more limited than the other groups. Virus shedding by SV581/11 group, however, lasted similarly or even longer than the other groups. Sequence analysis of three virulence genes (VEGF, VIR and IL-10v) revealed amino acid deletions and mutations in VEGF and IL-10v genes of SV581/11 and SV252/11, the isolate(s) producing milder lesions. Additionally, the VEGF gene of isolate SV581/11 presented the lowest amino acid identity with the other isolates and with ORFV standard strain OV-IA82. Thus, these results demonstrate that ORFV isolates may display differential virulence in lambs and these differences might be associated with genetic changes in putative virulence genes.

Diagnosis of contagious ecthyma in goats in a quarantine station in Panama / Diagnóstico de ectima contagioso en caprinos dentro de una estación de cuarentena en Panamá

We report an outbreak of contagious ecthyma (CE) in a herd of goats at Paso Canoas quarantine station, Panama. The goats were adult intact females. Visible clinical signs became apparent from day 13 after the start of quarantine. We performed clinical examination. Serum biopsy and scabs were collected from crusted lesions in the epithelium of the lips, nose and eyelid corners. Samples were studied by histopathology,complement fixation test, transmission electron microscopy (TEM), DAS-ELISA, viral isolationand nucleic acid amplification tests. Histopathology revealed ortho and parakeratotic hyperkeratosis, epithelial hyperplasia, viral inclusion bodies, keratinocytes with balonoid degeneration, vesicles with neutrophils and degenerated cells, in superficial dermis there is marked neovascularization. Complement fixation test, DAS-ELISA and nucleic acid amplification tests resulted positive for contagious ecthyma. TEM showed viral particles, consistent with Parapoxvirus. Clinical and laboratory findings were consistent with poxvirus infection in the quarantine goat herd.

El presente reporte describe un brote de ectima contagioso (EC) en un rebaño de cabras. Este caso tuvo lugar en la estación cuarentenaria de Paso Canoas, Panamá. Las cabras eran hembras, adultas, enteras y los signos clínicos fueron observados 13 días después de dar inicio al período de cuarentena. Se practicó el examen clínico, se colectaron fragmentos de costras y suero, además se realizaron biopsias de lesiones costrosas en el epitelio de los labios, nariz y comisuras palpebrales. Las muestras fueron analizadas por histopatología, prueba de fijación de complemento, microscopía electrónica de transmisión (MET), DAS-ELISA, aislamiento viral y amplificación de ácidos nucleicos. La histopatología reveló hiperqueratosis orto y paraqueratósica, hiperplasia epitelial, cuerpos de inclusión viral, eosinofílicos intracitoplasmáticos, degeneración balonoide de los queratinocitos, así como vesículas que contenían neutrófilos y células degeneradas; además, en la dermis superficial se observó una marcada neovascularización y edema. Las pruebas de fijación de complemento, DAS-ELISA y amplificación de ácidos nucléicos resultaron positivas para EC. El resultado de MET reveló partículas virales consistentes con Parapoxvirus. Los hallazgos clínicos y los resultados de laboratorio confirmaron el brote infeccioso de Parapoxvirus, agente etiológico de EC, en el hato en cuarentena.

Inactivated Parapoxvirus ovis induces a transient increase in the expression of proinflammatory, Th1-related, and autoregulatory cytokines in mice.

The immunostimulatory properties of inactivated Parapoxvirus ovis (iPPVO) have long been investigated in different animal species and experimental settings. In this study, we investigated the effects of iPPVO on cytokine expression in mice after intraperitoneal inoculation. Spleen and sera collected from iPPVO-treated mice at intervals after inoculation were submitted to cytokine mRNA determination by real-time PCR (qPCR), serum protein concentration by ELISA, and interferon (IFN)-α/ß activity by bioassay. The spleen of iPPVO-treated animals showed a significant increase in mRNA expression of all cytokines assayed, with different kinetics and magnitude. Proinflammatory cytokines interleukin (IL)-1ß, tumor necrosis factor-alpha (TNF-α), and IL-8 mRNA peaked at 24 hours postinoculation (hpi; 5.4-fold increase) and 48 hpi (3- and 10-fold increases), respectively. A 15-fold increase in IFN-γ and 6-fold IL-12 mRNA increase were detected at 48 and 24 hpi, respectively. Increased expression of autoregulatory cytokines (Th2), mainly IL-10 and IL-4, could be detected at later times (72 and 96 hpi) with peaks of 4.7- and 4.9-fold increases, respectively. IFN-I antiviral activity against encephalomyocarditis virus was demonstrated in sera of treated animals between 6 and 12 hpi, with a >90% reduction in the number of plaques. Measurement of serum proteins by ELISA revealed increased levels of IL-1, TNF-α, IL-12, IFN-γ, and IL-10, with kinetics similar to those observed by qPCR, especially for IL-12 and IFN-γ. These data demonstrate that iPPVO induced a transient and complex cytokine response, initially represented by Th1-related cytokines followed by autoregulatory and Th2 cytokines.

Inactivated Parapoxvirus ovis induces a transient increase in the expression of proinflammatory, Th1-related, and autoregulatory cytokines in mice

The immunostimulatory properties of inactivated Parapoxvirus ovis (iPPVO) have long been investigated in different animal species and experimental settings. In this study, we investigated the effects of iPPVO on cytokine expression in mice after intraperitoneal inoculation. Spleen and sera collected from iPPVO-treated mice at intervals after inoculation were submitted to cytokine mRNA determination by real-time PCR (qPCR), serum protein concentration by ELISA, and interferon (IFN)-α/β activity by bioassay. The spleen of iPPVO-treated animals showed a significant increase in mRNA expression of all cytokines assayed, with different kinetics and magnitude. Proinflammatory cytokines interleukin (IL)-1β, tumor necrosis factor-alpha (TNF-α), and IL-8 mRNA peaked at 24 hours postinoculation (hpi; 5.4-fold increase) and 48 hpi (3- and 10-fold increases), respectively. A 15-fold increase in IFN-γ and 6-fold IL-12 mRNA increase were detected at 48 and 24 hpi, respectively. Increased expression of autoregulatory cytokines (Th2), mainly IL-10 and IL-4, could be detected at later times (72 and 96 hpi) with peaks of 4.7- and 4.9-fold increases, respectively. IFN-I antiviral activity against encephalomyocarditis virus was demonstrated in sera of treated animals between 6 and 12 hpi, with a >90% reduction in the number of plaques. Measurement of serum proteins by ELISA revealed increased levels of IL-1, TNF-α, IL-12, IFN-γ, and IL-10, with kinetics similar to those observed by qPCR, especially for IL-12 and IFN-γ. These data demonstrate that iPPVO induced a transient and complex cytokine response, initially represented by Th1-related cytokines followed by autoregulatory and Th2 cytokines.

Partial sequence analysis of B2L gene of Brazilian orf viruses from sheep and goats.

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.

Looking back: a genetic retrospective study of Brazilian Orf virus isolates.

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.

Primeiro relato do vírus Orf em rebanhos caprinos do semiárido brasileiro / First report on the Orf virus infection in goat herds from Brazilian semiarid region

O Ectima contagioso, popularmente conhecido como Boqueira, é uma doença causada pelo vírus Orf, que induz lesões pustulares agudas na pele de ovinos e caprinos. No presente trabalho, é relatada pela primeira vez a identificação e confirmação laboratorial do vírus Orf em rebanhos caprinos da Bahia. A Agência Estadual de Defesa Agropecuária da Bahia (ADAB) relatou a ocorrência de focos de uma doença epitelial nos caprinos das raças Canindé e Alpina Britânica nas localidades de Barreira e Pedra Alta, no município de Araci. Foram coletadas as amostras das lesões de crostas labiais do rebanho da localidade de Barreira e das crostas dos tetos do rebanho de Pedra Alta. Os materiais coletados foram submetidos à extração de DNA e posterior reação em cadeia da polimerase (PCR), para amplificação dos genes do vírus Orf: ORFV011 (B2L, 1022 pares de bases [pb]) e ORFV 059 (F1L, 1062 pb). Todas as amostras foram positivas na reação de PCR, confirmando-se a presença do vírus Orf nas lesões observadas nos rebanhos caprinos das comunidades de Barreira e Pedra Alta.

Isolamento, avaliação do comportamento de amostras de vírus ectima contagioso e melhoramento de técnicas de diagnóstico e de cultivo in vitro

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.

Multifocal cutaneous ORF virus infection in goats in the Amazon region, Brazil.

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.