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In this study we report for the first time the phylodynamics of the parapoxvirus (PPV) genus in Mexico. Based on the analysis by PCR of 124 epithelial samples collected between 2007 and 2011 from naturally infected goats, sheep and cows in Mexico, we found that different PPV were present in 21 out of the 24 states sampled during this study. Our phylogenetic analysis confirmed the presence of different PPV species in Mexico, and their phylogenetic relationship with other PPV circulating in the US and Canada. Furthermore, we describe the existence of two different ORFV phylogenetic groups that are clearly host associated (sheep or goat). Evidence of directional selection at five specific amino acid residues in the enveloped glycoprotein B2L might help to support this host predilection. Collectively, the results generated in this study highlight the importance of PPV genus in Mexico and open the possibility for future studies describing with more detail the importance of this genus in North America.

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Orf virus (ORFV) is the etiological agent of Contagious Ecthyma (CE) disease that mainly affects sheep, goats, wild ruminants, and humans with a worldwide distribution. To date, only two strains from Argentinian sheep have been characterized at the molecular level and there is little information on ORFV strains circulating in Argentina. Here we describe and analyze five outbreaks of CE in goats in three geographic regions of the country: Northwest, Center, and Southwest. The phylogenetic analysis based on four molecular markers of ORFV (orf011 partial sequence and orf020, orf109, and orf127 complete sequence genes) revealed that there are different strains circulating in Argentina and pointed out the importance of knowing the health status of animals traded between farms.

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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.

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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.

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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.

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A enfermidade ectima contagioso está difundida em todo o estado de São Paulo. Foram amostrados 42 (8,64%) cuidadores de animais e 444 (91,36%) ovinos (n=486). A prevalência de reagentes para vírus-neutralização foi de 67% (IC95%=62-71%) nos ovinos, e em seus cuidadores de 76% (IC95%=63-89%), sendo P=0,22, ou seja, não houve diferença estatística significativa entre as espécies. A distribuição dos títulos teve diferença estatística significativa entre as espécies, com P=0,0048. As variações de titulação foram de 0,6 a 2,1 tanto nos ovinos quanto nos seus cuidadores. Dentre os 42 cuidadores de ovinos participantes do estudo, 32 apresentaram títulos de anticorpos expressos por log10 acima de 0,6.(AU)

These diseases are all widespread in the State of São Paulo. 42 (8.64%) animal caregivers and 444 (91.36%) sheep (n=486) were sampled. The reagents Prevalence paragraph virus neutralization was 67% (95% CI = 62-71%) in sheep and 76% (95% CI = 63-89%) for caregivers, with P=0.22 not being a statistically significant difference between the species. One of the distribution titles had significant difference between statistics as species with P=0.0048. The titration variations were 0.6 to 2.1, both in sheep and their caregivers. Among the 42 sheep caregivers participating in the study, 32 had antibody securities denominated in log10 above 0.6.(AU)

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A enfermidade ectima contagioso está difundida em todo o estado de São Paulo. Foram amostrados 42 (8,64%) cuidadores de animais e 444 (91,36%) ovinos (n=486). A prevalência de reagentes para vírus-neutralização foi de 67% (IC95%=62-71%) nos ovinos, e em seus cuidadores de 76% (IC95%=63-89%), sendo P=0,22, ou seja, não houve diferença estatística significativa entre as espécies. A distribuição dos títulos teve diferença estatística significativa entre as espécies, com P=0,0048. As variações de titulação foram de 0,6 a 2,1 tanto nos ovinos quanto nos seus cuidadores. Dentre os 42 cuidadores de ovinos participantes do estudo, 32 apresentaram títulos de anticorpos expressos por log10 acima de 0,6.(AU)

These diseases are all widespread in the State of São Paulo. 42 (8.64%) animal caregivers and 444 (91.36%) sheep (n=486) were sampled. The reagents Prevalence paragraph virus neutralization was 67% (95% CI = 62-71%) in sheep and 76% (95% CI = 63-89%) for caregivers, with P=0.22 not being a statistically significant difference between the species. One of the distribution titles had significant difference between statistics as species with P=0.0048. The titration variations were 0.6 to 2.1, both in sheep and their caregivers. Among the 42 sheep caregivers participating in the study, 32 had antibody securities denominated in log10 above 0.6.(AU)

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Abstract Milker's nodule is an occupational viral skin disease of universal distribution, caused by the Paravaccinia virus and that occurs in individuals who deal with dairy cattle herds. We describe a case acquired due to lack of use of PPE (Personal Protective Equipment) and perform a literature review.

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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.

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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.

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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: ORFV 011 (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.(AU)

The contagious ecthyma is a disease caused by Orf virus, which induces acute skin pustules lesions in sheep and goats. In Brazil the frequency of Orf virus infection seems to be higher than that officially reported. This study is the first report on the Orf virus identification and laboratory confirmation in herds of goats in Bahia. The State Agency of Agriculture Defense of Bahia reported the occurrence of an epithelial disease in goats of Canindé and British Alpine breeds in the Barreira and Pedra alta localities. The lesions were characterized by the occurrence of scabs around the labial fold or being restricted to the udders. The samples from lesions scab were collected from goats labial fold and teats. DNA samples were extracted from these materials, and analyzed by polymerase chain reaction (PCR), and the genes ORFV 011 (B2L, 1022 base pairs [bp]) and ORFV 059 (F1L, 1062 bp) were amplified. All of the samples showed positive PCR reaction, confirming the Orf virus infection in the goat herds of Bahia state.(AU)

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