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.

Rapid selection of recombinant orf virus expression vectors using green fluorescent protein / 南方医科大学学报

<p><b>OBJECTIVE</b>To construct a universal, highly attenuated orf virus expression vector for exogenous genes using green fluorescent protein (GFP) as the reporter gene.</p><p><b>METHODS</b>The flanking regions of the ORFV132 of orf virus DNA were amplified by PCR to construct the shuttle plasmid pSPV-132LF-EGFP-132RF. The shuttle plasmid was transfected into OFTu cells and GFP was incorporated into orf virus IA82Delta 121 by homologous recombination. The recombinant IA82Delta121-V was selected by green fluorescent signal. The deletion gene was identified by PCR and sequencing. The effects of ORFV132 knockout were evaluated by virus titration and by observing the proliferation of the infected vascular endothelial cells in vitro.</p><p><b>RESULTS</b>The recombinant orf virus IA82Delta121-V was obtained successfully and quickly, and the deletion of ORFV132 did not affect the replication of the virus in vitro but reduced its virulence.</p><p><b>CONCLUSION</b>Green fluorescent protein is a selectable marker for rapid, convenient and stable selection of the recombinant viruses. Highly attenuated recombinant orf virus IA82Delta121-V can serve as a new expression vector for exogenous genes.</p>

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/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.

Isolation and characterization of orf viruses from Korean black goats

Five cases of orf virus infection in Korean black goats were diagnosed in our laboratory between 2010 and 2011. One orf virus (ORF/2011) was isolated from an ovine testis cell line (OA3.Ts) for use as a vaccine candidate. Sequences of the major envelope protein and orf virus interferon resistance genes were determined and compared with published reference sequences. Phylogenetic analyses revealed that orf viruses from Korean black goats were most closely related to an isolate (ORF/09/Korea) from dairy goats in Korea. This result indicates that the orf viruses might have been introduced from dairy goats into the Korean black goat population.

Molecular mechanism of ORFV intervention strategies based on the UPS of host cell: a review / 病毒学报

In order to compete the antiviral effects of the host cell in the process of infection, ORFV(known as Orf virus) relies on a series of functional genes developed through long-term population evolution, such as interferon resistance genes, Bcl-2 protein genes and cell cycle inhibitor gene and so on, with these weapons this virus is able to effectively counteract immune clearance and immune regulation from a host cell. Concurrently, ORFV also focuses on exploiting signal transduction pathways of the ubiquitin-proteasome system(UPS), circumvents the intracellular signal transduction and CD8+ T activation, for shielding virus particles towards maturation and releasing outside. This review introduced inner link between the UPS of host cell and intervention mechanism by virus, and analyzed the key roles of certains components in UPS, these all together showed the evolution tendency of ORFV that was involved in the designing of inhibition to immune response and for intracellular immune escape upon the selection pressure in host cell infected.

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.

Molecular characteristics and immune evasion strategies of ORFV: a review / 病毒学报

Contagious ecthyma (also known as orf) is an acute skin zoonosis caused by orf virus (ORFV), which affects sheep, goats and humans. As one of the typical species of the Parapoxvirus genus of the Poxviridae family, orf virus has distinctive and unique characteristics of these species. A range of immuno-modulatory/pathogenesis -related genes acquired by virus that function is to limit (at least transiently) the effectiveness of host immunity during its evolution. This review is aimed to describe the latest progress on the molecular characteristics of ORFV, and upon which we analyzed molecular mechanism of the immune escape designed and a set of strategies developed for ORFV to effective against immune clearance of the host. Known as an essential component in evolutionary system, host is regulated by ORFV for using in population evolution. By the ORFV evolutional immune regulation components and its effect approach, we can understand the viral biological characteristics of ORFV, and it is helpful for us to further study the counter-measures of this disease.

Estandarización de una técnica molecular para el diagnóstico de virus orf útil en la ganadería ovina / Standardization of a molecular technique for the diagnosis of orf virus, useful in sheep raising

Orf o Ectima contagioso, enfermedad producida por virus DNA perteneciente a la familia poxviridae género parapoxvirus; se caracteriza por producir enfermedad vesicular en ganado, principalmente en rumiante menores. Personas que tengan contacto con animales u objetos contaminados con el virus pueden desarrollar la enfermedad, la cual es catalogada como la zoonosis de mayor diagnóstico en países donde la industria ovina se desarrolla a gran escala. Las lesiones más comunes se ubican en la zona bucal y ventanas nasales de animales y en dedos y manos en ser humano. En Chile existe la enfermedad y se han reportado casos en personas relacionadas a la industria ovina. No existe en el país método de diagnóstico, por lo cual se estandarizó una prueba de diagnóstico molecular (PCR) a partir de un oligonucleótido sintetizado y posteriormente se reprodujo la técnica a partir de un especimen biológico clínicamente positivo.

Orf or contagious Ectima, a disease produced by virus DNA belonging to the family poxviridae genus parapoxvirus is characterized by generating vesicular disease in cattle, specially in minor ruminants. Persons having contact with virus contaminated animals or objects might develop the disease, which is classified as the most frequently diagnosed zoonosis in countries where the sheep industry is developed on large scale. Most common lesions are located in the mouth zone and nasal windows of animals and in fingers and hands in the human being. The disease is present in Chile and cases related to persons working in the sheep industry have been reported. There is not a diagnosis method in the country, for which reason a molecular diagnosis test (PCR) was standardized from a synthesized oligonucleotid and subsequently the technique was reproduced from a clinically positive biological specimen.

Chronic and acute natural sheep orf infection: comparative clinico-patholoqical observations

This study involved clinico-pathological comparison of natural chronic and acute forms of orf virus infection which took place in a flock of sheep. Clinically, lesions in the acute orf took 5-6 weeks to heal, while healing in the chronic form took more than six months. Significant differences were observed on the histopathological lesions of both forms of the disease. Grossly, the scabs on the acute disease could be removed easily. However, in the chronic form they were strongly adherent to the skin and when forcibly removed they caused bleeding. Comparative discussion of both conditions was undertaken

Ectima contagioso: una enfermedad ocupacional en la industria ovina / Contagious ecthyma: an occupational disease in the sheep industry

Ectima contagioso, enfermedad producida por virus perteneciente a la familia poxviridae; se caracteriza por producir enfermedad vesicular en ganado, principalmente en rumiante menores. Personas que tengan contacto con animales u objetos contaminados con el virus pueden desarrollar la enfermedad, la cual es catalogada como la zoonosis de mayor diagnóstico en países donde la industria ovina se desarrolla a gran escala. Chile apunta al bicentenario triplicando la masa ovina existente en el país, por lo mismo, es necesario tomar en cuenta esta enfermedad que podría constituirse como problema en la producción ovina y en la salud ocupacional. Se analiza la posibilidad de realizar algunas pruebas de laboratorio para el diagnóstico precoz de la enfermedad y así estar preparados para los nuevos escenarios que se presenten.

Contagious ecthyma, a disease produced by a virus of the poxviridae family, characterized by producing vesicular disease in cattle, specially in small ruminants. People having contact with animals or objects contaminated with the virus may develop the disease, whichis classified as the principal diagnosed zoonosis, in countries where the sheep industry is developed in large scale. Chile aims at tripling the sheep mass existing in the country for the bicentennial. For this reason it is necessary to take into account that this disease might become a problem in sheep production and occupational health. The possibility of making some laboratory tests for the early diagnosis of the disease is discussed and thus be prepared to meet the new scenarios that might be encountered.

Orf: report of eleven cases in five Iranian families

Parapox virus is the causative agent of Orf disease which is basically seen in sheep and goats. It can be transferred to the human by direct contact with infected animals or indirect contact to infected meat or contaminated butchery instruments like knife. The skin lesions are usually found in fingers. Totally, 11 subjects from five families infected with Orf disease were surveyed. The diagnosis was verified on clinical manifestations and pathological changes. Local antiseptic [betadin] and tetraeycline ointment were administered for all patients, however, 3 cases were treated with oral antibiotic. No disseminated signs or complications have been reported and patients cured in the third [6 cases] or forth week [5 cases]. Little scars were noted in 3 cases alter one month follow up. If there is a similar lesion in the members of a family, one should notice the background of epidemiological and clinical manifestations. Despite the rarity of Orf disease, its spread still exists in urban families, therefore, it is essential to wear gloves at the time of touching meat to prevent infection

Haemophillus influenzae type b: molecular biological characteristics and familial contagious source

Haemophilus influenzae is one of the main pathogens of encephalitis in children less than 5 years old. The rate of ampicillin resistant strain is not high. No beta-lactamase-producing and ampicillin-resistant (BLNAR) isolates were found. The PFGE patterns of Hib isolates were highly divergent, but most could be classified into three groups (A, B and C). Hib isolates from the CSF of patients and from nasopharynges of household contacts showed the same PFGE patterns. This observation suggested that household contacts of patients are a possible reservoir of Hib.

Caracterización antigénica de cepas del virus del ectima contagioso (orf), sus interacciones y sus relaciones con parapoxvirus bovinos de México / Antigenic characterization of contagious ecthyma virus (orf) in Mexico, their relationships and interactions with bovine parapoxvirus samples

Treinta y tres muestras de costra positivas a ectima contagioso (orf) (19 de caprino y 14 ovinas) y dos de parapoxvirus bovinos (una de estomatitis papular y otra de seudoviruela) fueron analizadas antigénicamente y comparadas por doble inmunodifusión (DD) y contrainmunoelectroforesis (CIE). Las muestras bovinas fueron obtenidas de animales que convivían con rebaños de borregos y cabras. La presencia de partículas virales fue confirmada por microscopía electrónica en todos los casos. Las pruebas fueron corridas usando un suero policlonal antiectima preparado en conejo. En la prueba de DD la muestra homóloga produjo tres líneas de precipitación, las otras muestras variaron entre dos y una líneas de precipitación y algunas muestras no formaron ninguna, se observaron diferentes resultados con las pruebas de DD y CIE. Las cepas bovinas formaron dos líneas de precipitación, una de identidad entre ellas y otra en identidad con la muestra homóloga de orf caprino y con una de orf ovino, por DD. La composición de 15 muestras de orf (6 ovinas y 9 caprinas) y dos bovinas fueron examinadas por electroforesis, y 7 de ellas evaluadas antigénicamente por inmunotransferencia, usando el suero policlonal de conejo. La muestra homóloga fue más compleja, pues mostró 12 proteínas. La mayor variación en los corrimientos electroforéticos de las muestras se observó entre las proteínas de los pesos de 37 kd a 44 kd. Una proteína de 55 kd se identificó en todas las muestras, mientras que una de 9.5 kd se evidenció en 13 muestras y otra de 17 kd en 12, incluyendo las bovinas. Por inmunotransferencia fueron reconocidas dos proteínas comunes en las siete muestras analizadas con pesos de 55 y 54 kd, otras dos proteínas de 45 y 35 kd fueron reconocidas por el suero de conejo en cinco de las muestras. Estas proteínas pueden explicar las reacciones cruzadas observadas entre las muestras, en las pruebas de DD y CIE y ser de importancia en el reconocimiento inmune de estos virus.

Controle clínico de pacientes acometidos por infecções superficiais recorrentes pelo Staphylococcus aureus / Clinical control of patients attacked by superficial recurrent infections by Staphylococcus aureus

As infecções epidérmicas superficiais em pacientes portadores do Staphylococcus aureus na cavidade nasal representam um problema clínico relativamente comum, mas o seu manejo continua um desafio. Foi realizado um estudo descritivo, entre janeiro/94 e janeiro/96, com 60 pacientes de 18 a 70 anos de idade, em nível ambulatorial, onde foram avaliadas a eficácia terapêutica e a tolerância da associação rifampicina, mupirocina e gluconato de clorexidina a 2% no tratamento e controle dos portadores do Staphylococcus aureus. O resultado obtido mostrou que os paciente que completaram o esquema terapêutico proposto obtiveram melhora clínica e bacteriológica significativas (96% e 87%, respectivamente), onde conclui-se que a associação desses medicamentos apresenta-se como uma alternativa a ser considerada.

The epidermics superficials infections in patients carriers by Staphylococcus aureus in the cavit nasal represents a clinic problems relatively common, but that your management continue a challenge. A descriptive study was realized, between January 94/January 96, in 60 patients of 18-70 years old, at ambulatory level, where evaluated the eficacy and tolerance therapeutic of drug association rifampicina 300mg, mupirocin 2% and clorexidin 2%, in treatment and control of carriers to Staphylococcus aureus by realizated of swab intranasal. The result obtained point out the patients that concluded plan therapeutic proposed, got improvement clinic and bacteriology significant (96% and 87%, respectively) where conclude that drug association may be an alternative to be considered.

Pox del ganadero / Pox cattleman

Se han descrito en la literatura dos entidades, ambas provocadas por Poxvirus relacionados, que no pueden ser diferenciadas ni en bases clínicas ni histopatológicas. a ellas se las agrupan bajo el nombre de pox del ganadero; incluyendo así a la Enfermedad de Orf y nódulo del ordeñador. La enfermedad de Orf es endémica en ganado ovino y caprino, mientras que el nódulo del ordeñador se presenta principalmente en el ganado vacuno; ambas son de distribución mundial y por lo general de presentación esporádica, aunque se registran algunas epidemias aisladas. Estos virus pueden ser transmitidos en ambas enfermedades a través del contacto con animales infectados, ya sean vivos o muertos, o a través de objetos contaminados, debido a la resistencia de los virus al calor, frio o desecación. nosotros describimos un caso clínico de pox del ganadero incluyendo sus hallazgos histopatológicos.

ORF: Un Brote en Cabras Importadas / ORF: An Outbreak Among Imported Goats

Se describe un brote de orf en 94 cabras importads de EUA, destinadas a Lebrija y Floridablanca (Santander). Enfermaron 38 animales. No se presentaron complicaciones ni mortalidad. El diagnostico clinico se confirmo por la histopatologia, cultivo del virus en huevos embrionados y microscopia electronica con tincion negativa del macerado de las costras. No se comprobaron casos de infeccion humana. Dos de seis vacas ordenadas por un empleado a cargo de las cabras, presentaron papulas en la ubre pero no se realizaron en ellas ni en el trabajador, estudios virologicos. Se revisaron las caracteristicas de esta zoonosis que dentro de la poblacion animal tiene una distribucion amplia en Colombia, asi como sus implicaciones en la medicina humana y veterinaria. Se enfatiza sobre la necesidad de comprobar el diagnostico clinico por metodos virologicos y de microscopia electronica