RESUMEN
It has been shown that Muscovy duck reovirus (MDRV) infection causes severe intestinal barrier damage and intestinal mucosal immune suppression. The health and balance of gut microbes is essential for the progression of intestinal infectious diseases. To investigate the interaction of MDRV, intestinal bacteria with host intestinal innate immunity, an MDRV contact-infection model was established in this study. High-throughput sequencing technology was used to sequence 16S rDNA and transcripts in ileal samples from experimental Muscovy ducklings. Our results suggest that intestinal opportunistic pathogens such as Streptococcus and Corynebacterium proliferated massively in MDRV-infected Muscovy ducklings. The body initiates antiviral and antibacterial immunity and actively fights the infection of gut microbes. The synthesis of peptidoglycan, lipopolysaccharide, and flagellin by intestinal bacteria activates the Toll-like receptor signaling pathway resulting in increased secretion of IFN-ß, IL-1ß, and IL-8. The RIG-I-like receptor signaling pathway is an important signaling pathway for the interaction between MDRV and the host. At the same time, we also observed that multiple genes in the JAK-STAT signaling pathway were significantly different. These genes are important targets for studying the immunosuppression caused by MDRV. In conclusion, we analyzed the interaction of MDRV, intestinal flora and host immune system during MDRV infection, which provides a basis for the further study on the mechanism of intestinal immunosuppression caused by MDRV.
Asunto(s)
Patos , Microbioma Gastrointestinal , Interacciones Microbiota-Huesped , Inmunidad Innata , Infecciones por Reoviridae , Animales , Microbioma Gastrointestinal/genética , Microbioma Gastrointestinal/inmunología , Interacciones Microbiota-Huesped/inmunología , Inmunidad Innata/genética , Inmunidad Innata/inmunología , Reoviridae/fisiología , Infecciones por Reoviridae/inmunología , Infecciones por Reoviridae/microbiología , Infecciones por Reoviridae/veterinaria , Infecciones por Reoviridae/virología , TranscriptomaRESUMEN
Gut microbiota could facilitate host to defense diseases, but fish-microbiota interactions during viral infection and the underlying mechanism are poorly understood. We examined interactions and responses of gut microbiota to grass carp reovirus (GCRV) infection in Ctenopharyngodon idellus, which is the most important aquaculture fish worldwide. We found that GCRV infection group with serious haemorrhagic symptoms (G7s) showed considerably different gut microbiota, especially with an abnormally high abundance of gram-negative anaerobic Cetobacterium somerae. It also showed the lowest (p < 0.05) alpha-diversity but with much higher ecological process of homogenizing dispersal (28.8%), confirming a dysbiosis of the gut microbiota after viral infection. Interestingly, signaling pathways of NOD-like receptors (NLRs), toll-like receptors (TLRs), and lipopolysaccharide (LPS) stimulation genes were significantly (q-value < 0.01) enriched in G7s, which also significantly (p < 0.01) correlated with the core gut microbial genera of Cetobacterium and Acinetobacter. The results suggested that an expansion of C. somerae initiated by GCRV could aggravate host inflammatory reactions through the LPS-related NLRs and TLRs pathways. This study advances our understanding of the interplay between fish immunity and gut microbiota challenged by viruses; it also sheds new insights for ecological defense of fish diseases with the help of gut microbiota.
Asunto(s)
Carpas/microbiología , Carpas/virología , Enfermedades de los Peces/virología , Microbioma Gastrointestinal , Orthoreovirus Mamífero 3/fisiología , Infecciones por Reoviridae/veterinaria , Animales , Bacterias/clasificación , Bacterias/genética , Bacterias/aislamiento & purificación , Enfermedades de los Peces/microbiología , Fusobacterias , Interacciones Huésped-Patógeno , Orthoreovirus Mamífero 3/clasificación , Orthoreovirus Mamífero 3/genética , Orthoreovirus Mamífero 3/aislamiento & purificación , Infecciones por Reoviridae/microbiología , Infecciones por Reoviridae/virologíaRESUMEN
Duck spleen necrosis disease (DSND) is an emerging infectious disease that causes significant economic loss in the duck industry. In 2018, a duck reovirus (named DRV/GX-Y7) and Salmonella indiana were both isolated from the spleens and livers of diseased ducks with DSND in China. The DRV/GX-Y7 strain could propagate in the Vero, LMH, DF-1 and DEF cells with obvious cytopathic effects. The genome of DRV/GX-Y7 was 23,418 bp in length, contained 10 dsRNA segments, ranging from 3959 nt (L1) to 1191 nt (S4). The phylogenetic analysis showed that the DRV/GX-Y7 strain was in the same branch with the new waterfowl-origin reovirus cluster, but was obviously far distant from the clusters of other previous waterfowl-origin reoviruses Muscovy duck reovirus (MDRV) and goose-origin reovirus (GRV), broiler/layer-origin reovirus (ARV) and turkey-origin reovirus (TRV). The RDP and SimPlot program analysis revealed that there were two potential genetic reassortment events in the M2 and S1 segments of the genome. In order to have a clear insight into the pathogenic mechanism of DRV/GX-Y7 and S. Indiana in clinical DSND, an infection experiment was further conducted by challenging commercial ducklings with the two isolates individually and with both. The results showed that DRV/GX-Y7 produced severe hemorrhagic and/or necrotic lesions in the immune organs (thymus, spleen, and bursae) of experimentally infected ducklings. And, that the co-infection of DRV/GX-Y7 and S. Indiana could greatly enhance the pathogenesis by increasing the morbidity and mortality in ducklings whose clinical symptoms and lesions were similar to the natural clinical DSND cases. In summary, the results suggested that the pathogen causing duck spleen necrosis was an emerging unique genetic reassortment strain of duck Orthoreovirus that was significantly different from any previously reported waterfowl-derived Orthoreovirus and the co-infection with the Salmonella isolate could increase the severity of the disease.
Asunto(s)
Enfermedades Transmisibles Emergentes/veterinaria , Patos/virología , Enfermedades de las Aves de Corral/microbiología , Enfermedades de las Aves de Corral/virología , Infecciones por Reoviridae/veterinaria , Salmonelosis Animal/virología , Factores de Edad , Animales , China , Coinfección/veterinaria , Enfermedades Transmisibles Emergentes/virología , Hígado/patología , Hígado/virología , Orthoreovirus Aviar/genética , Orthoreovirus Aviar/patogenicidad , Enfermedades de las Aves de Corral/fisiopatología , Virus Reordenados/genética , Infecciones por Reoviridae/microbiología , Salmonella/genética , Salmonella/patogenicidad , Índice de Severidad de la Enfermedad , Bazo/patología , Bazo/virologíaRESUMEN
Enteric viruses infect the mammalian gastrointestinal tract which is home to a diverse community of intestinal bacteria. Accumulating evidence suggests that certain enteric viruses utilize these bacteria to promote infection. While this is not surprising considering their proximity, multiple viruses from different viral families have been shown to bind directly to bacteria or bacterial components to aid in viral replication, pathogenesis, and transmission. These data suggest that the concept of a single virus infecting a single cell, independent of the environment, needs to be reevaluated. In this review, I will discuss the current knowledge of enteric virus-bacterial interactions and discuss the implications for viral pathogenesis and transmission.
Asunto(s)
Tracto Gastrointestinal/virología , Interacciones Microbianas , Microbiota , Virus , Animales , Tracto Gastrointestinal/microbiología , Interacciones Microbiota-Huesped/inmunología , Humanos , Evasión Inmune , Picornaviridae/crecimiento & desarrollo , Picornaviridae/patogenicidad , Infecciones por Picornaviridae/inmunología , Infecciones por Picornaviridae/microbiología , Infecciones por Picornaviridae/transmisión , Reoviridae/crecimiento & desarrollo , Reoviridae/patogenicidad , Infecciones por Reoviridae/inmunología , Infecciones por Reoviridae/microbiología , Infecciones por Reoviridae/transmisión , Retroviridae/crecimiento & desarrollo , Retroviridae/patogenicidad , Infecciones por Retroviridae/inmunología , Infecciones por Retroviridae/microbiología , Infecciones por Retroviridae/transmisión , Virosis/inmunología , Virosis/microbiología , Virosis/transmisión , Replicación Viral , Virus/crecimiento & desarrollo , Virus/patogenicidadRESUMEN
Enteric viruses encounter diverse environments as they migrate through the gastrointestinal tract to infect their hosts. The interaction of eukaryotic viruses with members of the host microbiota can greatly impact various aspects of virus biology, including the efficiency with which viruses can infect their hosts. Mammalian orthoreovirus, a human enteric virus that infects most humans during childhood, is negatively affected by antibiotic treatment prior to infection. However, it is not known how components of the host microbiota affect reovirus infectivity. In this study, we show that reovirus virions directly interact with Gram positive and Gram negative bacteria. Reovirus interaction with bacterial cells conveys enhanced virion thermostability that translates into enhanced attachment and infection of cells following an environmental insult. Enhanced virion thermostability was also conveyed by bacterial envelope components lipopolysaccharide (LPS) and peptidoglycan (PG). Lipoteichoic acid and N-acetylglucosamine-containing polysaccharides enhanced virion stability in a serotype-dependent manner. LPS and PG also enhanced the thermostability of an intermediate reovirus particle (ISVP) that is associated with primary infection in the gut. Although LPS and PG alter reovirus thermostability, these bacterial envelope components did not affect reovirus utilization of its proteinaceous cellular receptor junctional adhesion molecule-A or cell entry kinetics. LPS and PG also did not affect the overall number of reovirus capsid proteins σ1 and σ3, suggesting their effect on virion thermostability is not mediated through altering the overall number of major capsid proteins on the virus. Incubation of reovirus with LPS and PG did not significantly affect the neutralizing efficiency of reovirus-specific antibodies. These data suggest that bacteria enhance reovirus infection of the intestinal tract by enhancing the thermal stability of the reovirus particle at a variety of temperatures through interactions between the viral particle and bacterial envelope components.
Asunto(s)
Bacillus subtilis/fisiología , Enterocitos/virología , Escherichia coli K12/fisiología , Infecciones por Reoviridae/virología , Reoviridae/fisiología , Acetilglucosamina/análogos & derivados , Acetilglucosamina/metabolismo , Acetilglucosamina/toxicidad , Bacillus subtilis/metabolismo , Bacillus subtilis/ultraestructura , Bacillus subtilis/virología , Células CACO-2 , Endotoxinas/metabolismo , Endotoxinas/toxicidad , Enterocitos/efectos de los fármacos , Enterocitos/microbiología , Enterocitos/patología , Escherichia coli K12/metabolismo , Escherichia coli K12/ultraestructura , Escherichia coli K12/virología , Microbioma Gastrointestinal , Células HeLa , Calor , Humanos , Lipopolisacáridos/metabolismo , Lipopolisacáridos/toxicidad , Proteínas Luminiscentes/genética , Proteínas Luminiscentes/metabolismo , Microscopía Electrónica de Transmisión , Peptidoglicano/metabolismo , Peptidoglicano/toxicidad , ARN/metabolismo , Estabilidad del ARN/efectos de los fármacos , Proteínas Recombinantes/metabolismo , Reoviridae/química , Reoviridae/efectos de los fármacos , Reoviridae/patogenicidad , Infecciones por Reoviridae/metabolismo , Infecciones por Reoviridae/microbiología , Infecciones por Reoviridae/patología , Ácidos Teicoicos/metabolismo , Ácidos Teicoicos/toxicidad , Virión/química , Virión/patogenicidad , Virión/fisiología , Acoplamiento Viral/efectos de los fármacos , Internalización del Virus/efectos de los fármacos , Proteína Fluorescente RojaRESUMEN
Toll-like receptors (TLRs) play a critical role in the innate immune system. Although TLR18 is an important member of this family of receptors in fish, the role of the tlr18 gene in responses to pathogen infection is still unclear. In this study, we identified and characterized the grass carp tlr18 gene (gctlr18) to further clarify the function of TLR18 in teleost fish. Gctlr18 spans over 3600 bp and encodes a polypeptide of 852 amino acids. Analysis of the deduced amino acid sequence showed that gctlr18 encodes structures typical of the TLR family, including a signal peptide, seven leucine-rich repeats (LRRs), a transmembrane region, and a (Toll-interleukin-1 receptor) TIR domain. Quantitative RT-PCR analysis showed that gctlr18 was constitutively expressed in all investigated tissues, with abundant expression in spleen, gill, heart, intestine, kidney and fin and low expression in skin, liver and brain. Following grass carp reovirus-challenge and Aeromonas hydrophila inoculation, gctlr18 transcripts were upregulated significantly in immune-relevant tissues. Stimulation of Ctenopharyngodon idella kidney (CIK) cells with purified flagellin from Salmo typhimurium, lipopolysaccharide and polyinosinic-polycytidylic acid stimulation in vitro resulted in significantly increased gctlr18 expression, reaching a peak followed by restoration of normal levels. Overexpression of gctlr18 reduced A. hydrophila invasion by 83.4%. In CIK cells, gctlr18 induced the expression of proinflammatory cytokines, including il-8, inf-1 and tnf-α. Our results indicate that gctlr18 plays a key role in innate immune responses in teleost fish.
Asunto(s)
Carpas/genética , Enfermedades de los Peces/inmunología , Proteínas de Peces/genética , Regulación de la Expresión Génica , Infecciones por Bacterias Gramnegativas/veterinaria , Infecciones por Reoviridae/veterinaria , Receptores Toll-Like/genética , Aeromonas hydrophila/fisiología , Animales , Clonación Molecular , ADN Complementario/genética , ADN Complementario/metabolismo , Enfermedades de los Peces/genética , Enfermedades de los Peces/microbiología , Enfermedades de los Peces/virología , Proteínas de Peces/metabolismo , Infecciones por Bacterias Gramnegativas/genética , Infecciones por Bacterias Gramnegativas/inmunología , Infecciones por Bacterias Gramnegativas/microbiología , Datos de Secuencia Molecular , Filogenia , ARN Mensajero/genética , ARN Mensajero/metabolismo , Reoviridae/fisiología , Infecciones por Reoviridae/genética , Infecciones por Reoviridae/inmunología , Infecciones por Reoviridae/microbiología , Análisis de Secuencia de ADN/veterinaria , Receptores Toll-Like/metabolismoRESUMEN
As obligate intracellular parasites, viruses are exclusively and intimately dependent upon their host cells for replication. During replication viruses induce profound changes within cells, including: induction of signaling pathways, morphological changes, and cell death. Many such cellular perturbations have been analyzed at the transcriptomic level by gene arrays and recent efforts have begun to analyze cellular proteomic responses. We recently described comparative stable isotopic (SILAC) analyses of reovirus, strain type 3 Dearing (T3D)-infected HeLa cells. For the present study we employed the complementary labeling strategy of iTRAQ (isobaric tags for relative and absolute quantitation) to examine HeLa cell changes induced by T3D, another reovirus strain, type 1 Lang, and UV-inactivated T3D (UV-T3D). Triplicate replicates of cytosolic and nuclear fractions identified a total of 2375 proteins, of which 50, 57, and 46 were significantly up-regulated, and 37, 26, and 44 were significantly down-regulated by T1L, T3D, and UV-T3D, respectively. Several pathways, most notably the Interferon signaling pathway and the EIF2 and ILK signaling pathways, were induced by virus infection. Western blots confirmed that cells were more strongly activated by live T3D as demonstrated by elevated levels of key proteins like STAT-1, ISG-15, IFIT-1, IFIT-3, and Mx1. This study expands our understanding of reovirus-induced host responses.
Asunto(s)
Proteínas Portadoras/metabolismo , Citocinas/metabolismo , Interferones/metabolismo , Péptidos y Proteínas de Señalización Intracelular/metabolismo , Orthoreovirus Mamífero 3/fisiología , Proteínas de Resistencia a Mixovirus/metabolismo , Infecciones por Reoviridae/metabolismo , Factor de Transcripción STAT1/metabolismo , Ubiquitinas/metabolismo , Proteínas Adaptadoras Transductoras de Señales , Proteínas Portadoras/genética , Citocinas/genética , Células HeLa , Humanos , Interferones/genética , Péptidos y Proteínas de Señalización Intracelular/genética , Orthoreovirus Mamífero 3/genética , Proteínas de Resistencia a Mixovirus/genética , Proteómica , Proteínas de Unión al ARN , Infecciones por Reoviridae/genética , Infecciones por Reoviridae/microbiología , Factor de Transcripción STAT1/genética , Ubiquitinas/genéticaRESUMEN
Mycoplasma synoviae and avian reovirus (ARV) are associated with several disease syndromes in poultry and cause notable global economic losses in the poultry industry. Rapid and efficient diagnostics for these avian pathogens are important not only for disease control but also for prevention of clinical disease progression. However, current diagnostic methods used for surveillance of these diseases in poultry flocks are laborious and time-consuming, and they have low sensitivity. The multiplex PCR (mPCR) developed in this study has been proven to be both sensitive and specific for simultaneous M. synoviae and ARV detection and identification in clinical samples. To evaluate the mPCR assay, the diagnostic test was applied to different clinical samples from natural and experimental M. synoviae and ARV-infected poultry. Results were compared with serologic, single PCR, and immunofluorescence analyses. Tibiotarsal articulation could be the best target for simultaneous detection of M. synoviae and ARV infection. The detection limit by visualization of mPCR-amplified products was 100 pg for both pathogens. Overall, the mPCR developed and standardized in this research is a useful tool for diagnosis and screening and for surveillance and control of M. synoviae and ARV infection in poultry flocks.
Asunto(s)
Pollos , Reacción en Cadena de la Polimerasa Multiplex/métodos , Infecciones por Mycoplasma/veterinaria , Mycoplasma synoviae/genética , Orthoreovirus Aviar/genética , Enfermedades de las Aves de Corral/diagnóstico , Infecciones por Reoviridae/diagnóstico , Animales , ADN Bacteriano/genética , ADN Bacteriano/metabolismo , Femenino , Técnica del Anticuerpo Fluorescente/veterinaria , Reacción en Cadena de la Polimerasa Multiplex/veterinaria , Infecciones por Mycoplasma/diagnóstico , Infecciones por Mycoplasma/microbiología , Mycoplasma synoviae/aislamiento & purificación , Mycoplasma synoviae/metabolismo , Orthoreovirus Aviar/aislamiento & purificación , Orthoreovirus Aviar/metabolismo , Enfermedades de las Aves de Corral/microbiología , Infecciones por Reoviridae/microbiología , Sensibilidad y EspecificidadRESUMEN
Reovirus infection of target cells can perturb cell cycle regulation and induce apoptosis. Differences in the capacity of reovirus strains to induce cell cycle arrest at G1 and G2/M have been mapped to the viral S1 genome segment, which also determines differences in the ability of reovirus strains to induce apoptosis and to activate specific mitogen-activated protein kinase (MAPK) cascades selectively. Reovirus-induced apoptosis involves members of the tumor necrosis factor (TNF) superfamily of death receptors and is associated with activation of both death receptor- and mitochondrial-associated caspases. Reovirus infection is also associated with the activation of a variety of transcription factors, including nuclear factor (NF)-kappaB. Junctional adhesion molecule (JAM) has recently been identified as a novel reovirus receptor. Reovirus binding to JAM appears to be required for induction of apoptosis and activation of NF-kappaB, although the precise cellular pathways involved have not yet been identified.
Asunto(s)
Receptores Virales/metabolismo , Infecciones por Reoviridae/genética , Reoviridae/fisiología , Factores de Transcripción/metabolismo , Animales , Apoptosis , Moléculas de Adhesión Celular/metabolismo , Ciclo Celular , Interacciones Huésped-Parásitos , Moléculas de Adhesión de Unión , Ratones , Infecciones por Reoviridae/microbiología , Factores de Transcripción/genéticaRESUMEN
A panel of serotype 3 (T3) reovirus strains was screened to determine their relative capacities to cause lethal infection and hepatobiliary disease following peroral inoculation in newborn mice. A wide range of 50% lethal doses (LD50s) was apparent after peroral inoculation of the different virus strains. Two of the strains, T3 Abney and T3 clone 31, caused mice to develop the oily fur syndrome associated with biliary atresia. The capacity to cause biliary atresia was not related to the capacity to cause lethal infection, however, because the LD50s of T3 Abney and T3 clone 31 were grossly disparate. Examination of liver and bile duct tissues revealed histopathologic evidence of biliary atresia and hepatic necrosis in T3 Abney-infected mice but not in mice inoculated with a T3 strain of similar virulence or with the hepatotropic T1 Lang strain. The consistency with which T3 Abney-infected mice developed biliary atresia-associated oily fur syndrome permitted us to determine the viral genetic basis of reovirus-induced biliary atresia. Analysis of reassortant viruses isolated from an in vitro coinfection with T3 Abney and T1 Lang indicated a strong association of the hepatobiliary disease-producing phenotype with the T3 Abney S1 gene, which encodes the viral cell attachment protein, sigma 1. Amino acid residues within the sigma 1 protein that were unique to disease-producing T3 strains were identified by comparative sequence analysis. Specific changes exist within two regions of the protein, one of which is thought to be involved in binding to host cell receptors. We hypothesize that changes within this region of the protein are important in determining the tropism of this virus for bile-ductular epithelium.
Asunto(s)
Atresia Biliar/microbiología , Proteínas de la Cápside , Genes Virales , Orthoreovirus Mamífero 3/genética , Proteínas de Unión al ARN , Infecciones por Reoviridae/fisiopatología , Proteínas Virales/biosíntesis , Secuencia de Aminoácidos , Animales , Animales Recién Nacidos , Antígenos Virales/análisis , Atresia Biliar/patología , Encéfalo/microbiología , Encéfalo/patología , Bovinos , Humanos , Inmunohistoquímica , Células L , Hígado/microbiología , Hígado/patología , Orthoreovirus Mamífero 3/aislamiento & purificación , Orthoreovirus Mamífero 3/patogenicidad , Ratones , Ratones Endogámicos , Datos de Secuencia Molecular , Estructura Secundaria de Proteína , Infecciones por Reoviridae/microbiología , Infecciones por Reoviridae/patología , Homología de Secuencia de Aminoácido , Proteínas Virales/química , Proteínas Virales/genética , Virión/patogenicidadRESUMEN
The C-polyhedrin genes of two different geographic isolates of a type 5 cytoplasmic polyhedrosis virus (CPV) were cloned. A CPV infecting Orgyia pseudotsugata (OpCPV), isolated in the Pacific Northwest of the U.S.A., and a CPV infecting Heliothis armigera, isolated in South Africa, were studied. Both genes were found to be 883 nucleotides in length and encoded a predicted protein of 246 residues (M(r) of 28,890). Comparison of the nucleotide sequences of these two viruses with another type 5 geographic isolate, infecting Euxoa scandens (EsCPV; isolated in Eastern Canada), showed that there were only 17 nucleotide differences among the three genes. The only nucleotide variation that had an effect on the encoded protein was a deletion of nucleotide 774 in the gene of EsCPV. The deletion introduces a frameshift mutation resulting in the alteration of the carboxyl-terminal amino acid sequence. Sequence alignment of the OpCPV C-polyhedrin showed little homology to a type 1 CPV (infecting Bombyx mori) or with analogous proteins (N-polyhedrins) from two baculoviruses infecting O. pseudotsugata. Interestingly, most of the conserved residues between the N- and C-polyhedrins were either basic or aromatic amino acids.
Asunto(s)
Genes Virales/genética , Mariposas Nocturnas/microbiología , Infecciones por Reoviridae/microbiología , Reoviridae/genética , Proteínas Virales/genética , Secuencia de Aminoácidos , Animales , Secuencia de Bases , Canadá , Variación Genética , Datos de Secuencia Molecular , Noroeste de Estados Unidos , Proteínas de la Matriz de Cuerpos de Oclusión , Reoviridae/clasificación , Homología de Secuencia de Aminoácido , Sudáfrica , Especificidad de la Especie , Proteínas Virales/química , Proteínas Estructurales ViralesRESUMEN
The potential for the use of the polymerase chain reaction (PCR) in detecting epizootic hemorrhagic disease virus (EHDV) ribonucleic acid in cell cultures and clinical samples was studied. Using oligoribonucleotide primers selected from genome segment 6 of EHDV-2 (Alberta strain), the PCR-based assay resulted in a 387-base pair (bp) PCR product. EHDV RNA from US prototype serotypes 1 and 2 and a number of EHDV field isolates, propagated in cell cultures, were detected by this EHDV PCR-based assay. Amplification products were visualized on ethidium bromide-stained agarose gels or detected by chemiluminescent hybridization. The sensitivity of the PCR assay was 100 fg of virus RNA (equivalent to 6 x 10(3) virus particles) with ethidium bromide-stained agarose gels. Chemiluminescent hybridization increased the sensitivity of the PCR assay 1,000 times, and specific signals were detected from 0.1 fg of virus RNA (equivalent to 6 virus particles). Amplification product was not detected when the PCR-based assay was applied to RNA from the US bluetongue (BT) virus prototype serotypes 2, 10, 11, 13, and 17 or total nucleic acid extracts from uninfected baby hamster kidney-21 cells, Vero cells, and blood cells from deer that were EHDV seronegative and virus isolation negative. Application of this EHDV PCR-based assay to clinical samples resulted in detection of EHDV RNA from blood and spleen samples from a deer in California with clinical hemorrhagic disease. This EHDV PCR-based assay could provide a rapid, sensitive, and specific assay for detection of EHDV infection in susceptible ruminants.
Asunto(s)
Ciervos/microbiología , Virus de la Enfermedad Hemorrágica Epizoótica/aislamiento & purificación , ARN Viral/análisis , Infecciones por Reoviridae/veterinaria , Animales , Secuencia de Bases , Southern Blotting/veterinaria , Células Cultivadas , Cricetinae , Cartilla de ADN , Electroforesis en Gel de Agar/veterinaria , Virus de la Enfermedad Hemorrágica Epizoótica/clasificación , Datos de Secuencia Molecular , Reacción en Cadena de la Polimerasa/veterinaria , Infecciones por Reoviridae/diagnóstico , Infecciones por Reoviridae/microbiología , Reproducibilidad de los Resultados , Sensibilidad y Especificidad , Serotipificación/veterinaria , Células VeroAsunto(s)
Orbivirus/clasificación , Animales , Bovinos , Enfermedades de los Bovinos/microbiología , Línea Celular , Ceratopogonidae , Culex , Pruebas de Inhibición de Hemaglutinación , Japón , Pruebas de Neutralización , Orbivirus/inmunología , Infecciones por Reoviridae/microbiología , Infecciones por Reoviridae/veterinariaRESUMEN
A fluorescence inhibition test (FIT) is described for serotyping rapidly isolates of epizootic haemorrhagic disease of deer virus (EHDV). The test used a serogroup-reactive monoclonal antibody in a immunofluorescence procedure to detect virus which resisted neutralisation by antisera to any of the eight known EHDV serotypes. The EHDV FIT provided an accurate serotype identification procedure for all eight reference serotypes and, in comparison with the plaque inhibition assay, abbreviated the serotyping process by three to four days.
Asunto(s)
Ciervos , Técnica del Anticuerpo Fluorescente , Virus de la Enfermedad Hemorrágica Epizoótica/aislamiento & purificación , Infecciones por Reoviridae/veterinaria , Animales , Bovinos , Células Cultivadas , Cricetinae , Virus de la Enfermedad Hemorrágica Epizoótica/clasificación , Infecciones por Reoviridae/microbiología , Serotipificación , Especificidad de la Especie , Factores de Tiempo , Ensayo de Placa ViralRESUMEN
Experiments were undertaken to examine the chymotrypsin sensitivity and trypsin sensitivity of 13 avian reoviruses, and to determine if there was any correlation with pathogenicity of some chicken reoviruses. A wide variation in the degree of sensitivity of avian reoviruses to chymotrypsin and trypsin was observed. Overall, the infectivity of the 13 avian reoviruses for Vero cells was markedly reduced by treatment with 0.01% chymotrypsin (the lowest concentration tested) while 0.5% trypsin significantly reduced the infectivity of 9 of 13 strains. Comparison of four avian reoviruses, three resistant and one sensitive to trypsin, for pathogenicity in day old chicks following oral inoculation showed the strains that were resistant to trypsin to be more pathogenic. Tenosynovitis and virus persistence in intestines, liver, heart and hock joint tissues occurred only in chickens inoculated with the trypsin resistant strains. It is concluded that the degree of sensitivity to chymotrypsin and trypsin among avian reoviruses is heterogenous. Sensitivity to trypsin influenced the development of tenosynovitis based on microscopic lesions and virus persistence in tissues.
Asunto(s)
Enfermedades de las Aves , Pollos/microbiología , Quimotripsina/toxicidad , Infecciones por Reoviridae/veterinaria , Reoviridae/efectos de los fármacos , Reoviridae/patogenicidad , Tripsina/toxicidad , Animales , Antivirales/toxicidad , Embrión de Pollo , Cloaca/microbiología , Corazón/microbiología , Articulaciones/microbiología , Hígado/microbiología , Reoviridae/aislamiento & purificación , Infecciones por Reoviridae/microbiología , Infecciones por Reoviridae/patología , Células VeroRESUMEN
Essaouira and Kala Iris viruses were isolated from Ornithodoros (Alectorobius) maritimus ticks parasitizing yellow-legged gulls (Larus cachinnans) on the coast of Morocco in 1979 and 1981, respectively. Serological evidence indicates that these two viruses are new members of the Chenuda complex within the Kemerovo serogroup of the genus Orbivirus. Ecological, pathological, morphological, and physicochemical properties are compatible with these findings. The infectivity of these viruses for man and animals, including seabirds, remains unknown.
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Orbivirus/clasificación , Garrapatas/microbiología , Animales , Antígenos Virales/análisis , Aves/microbiología , Encéfalo/microbiología , Encéfalo/patología , Femenino , Masculino , Marruecos , Orbivirus/inmunología , Orbivirus/aislamiento & purificación , Orbivirus/patogenicidad , Orbivirus/ultraestructura , Infecciones por Reoviridae/microbiología , Serotipificación , VirulenciaRESUMEN
A panel of reovirus strains was used to compare myocarditic potential with induction of cytopathic effect in primary cardiac myocyte and cardiac fibroblast cultures. The results suggest that viral cytopathogenicity in cardiac myocytes, but not in cardiac fibroblasts, is a determinant of reoviral myocarditis.
Asunto(s)
Corazón/microbiología , Miocarditis/microbiología , Infecciones por Reoviridae/microbiología , Reoviridae/patogenicidad , Animales , Animales Recién Nacidos , Células Cultivadas , Fibroblastos/microbiología , Fibroblastos/patología , Ratones , Ratones Endogámicos , Miocarditis/patología , Miocardio/patología , Reoviridae/fisiología , Infecciones por Reoviridae/patología , Replicación ViralRESUMEN
Virus was isolated from the blood of horses (n = 5) showing fever and jaundice and was identified as equine encephalosis virus. In cross neutralisation tests, the isolates were shown to belong to a new serotype related to Gamil, one of the 6 known serotypes of equine encephalosis virus. The name Potchefstroom has been proposed for this new serotype.
Asunto(s)
Enfermedades de los Caballos/microbiología , Orbivirus/aislamiento & purificación , Infecciones por Reoviridae/microbiología , Animales , Femenino , Caballos , Orbivirus/clasificación , SerotipificaciónRESUMEN
A monoclonal antibody (mAb) to infectious bursal disease virus (IBDV) was bound to polystyrene latex microspheres. The microspheres agglutinated with extracts of bursae and sera from chickens infected with all strains or isolates of IBDV tested. Agglutination appeared within a 10-min reaction time. The assay could detect a 10(3.7) to 10(4.5) mean embryo infective dose (EID50) of the virus in 0.01 ml and the titer of the assay was 10- to 40-times higher than that of the agar gel precipitin test.
Asunto(s)
Anticuerpos Monoclonales/inmunología , Anticuerpos Antivirales/inmunología , Pollos/microbiología , Virus de la Enfermedad Infecciosa de la Bolsa/aislamiento & purificación , Pruebas de Fijación de Látex , Enfermedades de las Aves de Corral/microbiología , Infecciones por Reoviridae/veterinaria , Animales , Antígenos Virales/análisis , Bolsa de Fabricio/microbiología , Embrión de Pollo/microbiología , Pollos/sangre , Pollos/inmunología , Virus de la Enfermedad Infecciosa de la Bolsa/inmunología , Virus de la Enfermedad Infecciosa de la Bolsa/patogenicidad , Ratones , Ratones Endogámicos BALB C/inmunología , Microesferas , Enfermedades de las Aves de Corral/sangre , Enfermedades de las Aves de Corral/inmunología , Pruebas de Precipitina , Infecciones por Reoviridae/sangre , Infecciones por Reoviridae/inmunología , Infecciones por Reoviridae/microbiología , Sensibilidad y Especificidad , Factores de Tiempo , Vacunas Virales , Viremia/microbiología , Viremia/veterinariaRESUMEN
A nucleic acid hybridization assay was developed to rapidly detect small quantities of aquareovirus RNAs in infected cells and organs. Cloned cDNA copies were synthesized from the genomic RNA of the SBR strain of aquareovirus. By using cloned cDNA probes, aquareovirus RNAs were detected in spleen and kidney tissues of experimentally infected fish.