RESUMEN
Two defective bovine parainfluenza virus type 3 (BPIV3) strains were generated, one lacking the membrane (M) protein gene and expressing EGFP (ΔM-EGFP) and the other lacking the fusion (F) protein gene and expressing mStrawberry (ΔF-mSB), by supplying deficient proteins in trans. When Madin-Darby bovine kidney (MDBK) cells were co-infected with ΔM-EGFP and ΔF-mSB at a multiplicity of infection (MOI) of 0.1, complemented viruses were easily obtained. Complemented viruses grew as efficiently as wild-type BPIV3 and could be passaged in MDBK cell cultures even at an MOI of 0.01, possibly due to multiploid virus particles containing genomes of both ΔM-EGFP and ΔF-mSB. This reciprocal complementation method using two defective viruses would be useful to express large or multiple proteins in cell cultures using paramyxovirus vectors.
Asunto(s)
Virus Defectuosos/genética , Prueba de Complementación Genética , Virus de la Parainfluenza 3 Bovina/genética , Animales , Bovinos , Técnicas de Cultivo de Célula , Línea Celular , Prueba de Complementación Genética/instrumentación , Prueba de Complementación Genética/métodos , Vectores Genéticos , Virus de la Parainfluenza 3 Bovina/crecimiento & desarrollo , Proteínas Virales de Fusión/genética , Proteínas de la Matriz Viral/genética , Replicación ViralRESUMEN
The bovine parainfluenza virus type 3 BN-CE vaccine strain was obtained by serial passage of the BN-1 strain in chicken embryonic fibroblasts (CEF). We previously identified a substitution (L288I) in the fusion (F) protein between the two strains. To examine the effect of the substitution on CEF adaptation and attenuation, we generated a recombinant BN-1 strain with the L288I substitution in the F protein (FL288I-EGFP). FL288I-EGFP replicated more efficiently than a recombinant BN-1 strain (wt-EGFP) in semi-suitable cell lines, suggesting that the L288I substitution was established in the BN-1 strain during the process of adaptation in CEF.
Asunto(s)
Adaptación Fisiológica/genética , Sustitución de Aminoácidos , Virus de la Parainfluenza 3 Bovina/genética , Virus de la Parainfluenza 3 Bovina/fisiología , Proteínas Virales de Fusión/genética , Proteínas Virales de Fusión/fisiología , Animales , Bovinos , Línea Celular , Células HeLa , Humanos , Virus de la Parainfluenza 3 Bovina/crecimiento & desarrollo , Proteínas Virales de Fusión/química , Replicación ViralRESUMEN
An outbreak of contagious mastitis occurred among cattle on a farm, and bovine herpesviruses were isolated from the affected mammary tissues, scabs and abscess discharge of the cattle. A bovine herpesvirus type 4 (BoHV-4)-specific fragment was amplified from the isolates by polymerase chain reaction (PCR). Restriction endonuclease analyses demonstrated that the isolates were related to Movar-like European type BoHV-4. To determine the ratio of BoHV-4 subclinical infection in the cattle, a genomic survey was performed by PCR for cattle that were moved to the animal hygiene service station in Ibaraki prefecture. The BoHV-4 genome was occasionally detected in peripheral blood leukocytes, lymph nodes and nervous tissues. The rate of BoHV-4 subclinical infection was relatively high in the cattle.
Asunto(s)
Enfermedades de los Bovinos/virología , Infecciones por Herpesviridae/veterinaria , Herpesvirus Bovino 4/clasificación , Infecciones Tumorales por Virus/veterinaria , Absceso/veterinaria , Absceso/virología , Animales , Anticuerpos Antivirales/sangre , Bovinos , Efecto Citopatogénico Viral , ADN Viral/química , ADN Viral/genética , Femenino , Infecciones por Herpesviridae/virología , Herpesvirus Bovino 4/genética , Herpesvirus Bovino 4/crecimiento & desarrollo , Herpesvirus Bovino 4/aislamiento & purificación , Reacción en Cadena de la Polimerasa/veterinaria , Polimorfismo de Longitud del Fragmento de Restricción , Infecciones Tumorales por Virus/virologíaRESUMEN
A disease characterized by arthritis of carpal joints and occasionally by pneumonia was seen among goats on a farm in Nagano prefecture of Japan in the summer of 2002. Serological investigation was done on 30 goats, that is one group on the farm by agar gel immunodiffusion tests using maedi-visna virus as the antigen, and 19 goats were positive. The caprine arthritis encephalitis virus genome was detected in peripheral blood leukocytes from several antibody-positive goats. Two goats among them were necropsied and used for pathological tests and virus isolation. The isolated virus was serially propagated in fetal lamb lung cell cultures. Goats inoculated with the culture fluid became antibody positive. These results suggested that a CAEV-infected goat had been introduced accidentally to the farm and that subclinical infection occurred among the flock.
Asunto(s)
Virus de la Artritis-Encefalitis Caprina/genética , Brotes de Enfermedades , Enfermedades de las Cabras/epidemiología , Enfermedades de las Cabras/virología , Infecciones por Lentivirus/veterinaria , Animales , Secuencia de Bases , Células Cultivadas/ultraestructura , Cartilla de ADN , Técnica del Anticuerpo Fluorescente/veterinaria , Enfermedades de las Cabras/patología , Cabras , Inmunodifusión , Japón/epidemiología , Infecciones por Lentivirus/epidemiología , Infecciones por Lentivirus/patología , Microscopía Electrónica/veterinaria , Datos de Secuencia Molecular , Reacción en Cadena de la Polimerasa/veterinaria , Alineación de Secuencia , Análisis de Secuencia de ADN/veterinariaRESUMEN
Intranuclear eosinophilic inclusion bodies were seen in the lactiferous duct and sinus epithelium of mammary tissues collected from a cow with clinical mastitis. Transmission electron microscopy revealed herpesvirus particles in these cells. Immunolabeling against anti bovine herpesvirus type 4 (BHV-4) rabbit serum was detected in nuclei that had intranuclear inclusion bodies. In addition, BHV-4 was isolated from the mammary tissue. The viral DNA was detected by nested PCR from the same tissue. This is the first report to describe mammary lesions in association with BHV-4.