RESUMO
Epizootic hemorrhagic disease virus (EHDV), like other orbiviruses, infects and replicates in mammalian and insect vector cells. Within its ruminant hosts EHDV, like bluetongue virus (BTV), it has mainly been associated with infection of endothelial cells of capillaries as well as leukocyte subsets. Furthermore, EHDV infects and replicates within its biological vector, Culicoides biting midges and Culicoides-derived cells. A wide range of common laboratory cell lines such as BHK, BSR, and Vero cells are susceptible to infection with certain EHDV strains. Cell culture supernatants of infected cells are commonly used for both in vivo and in vitro infection studies. For specific virological or immunological studies, using highly purified virus particles, however, might be beneficial or even required. Here we describe a purification method for EHDV particles, which had been originally developed for certain strains of BTV.
Assuntos
Vírus da Doença Hemorrágica Epizoótica , Vírion , Animais , Vírus da Doença Hemorrágica Epizoótica/isolamento & purificação , Linhagem Celular , Vírion/isolamento & purificação , Chlorocebus aethiops , Células Vero , Orbivirus/isolamento & purificação , Ceratopogonidae/virologia , Insetos/virologia , Infecções por Reoviridae/virologia , Infecções por Reoviridae/veterinária , CricetinaeRESUMO
The virus neutralization test (VNT) is a functional immunoassay which detects the presence and quantity of neutralizing antibodies. It is a highly sensitive and specific test. As with most neutralization assays, the EHDV VNT does not react with all virus-targeting antibodies, but specifically with those antibodies that bind to VP2, the outermost capsid structural protein of the virus. The interaction between VP2 and neutralizing antibodies can block EHDV cell binding, neutralizing its infectivity. The detection and quantification of neutralizing antibodies are indicative of how protected an animal is against reinfection. The EHD VNT can therefore be a useful tool to monitor the efficacy of a vaccination campaign. VP2 is also the main determinant of EHDV serotype specificity, and so EHDV-neutralizing antibodies which target VP2 are also serotype-specific. Throughdetecting and quantifying neutralizing antibodies, the VNT can discriminate the EHDV serotype responsible for an infection and provides insights into the time of infection. It is considered the gold standard test for identifying and quantifying antibodies against EHDV serotypes present in test serum samples. The assay is performed in vitro and is based on inhibition of virus infectivity in the presence of neutralizing antibodies. A neutralizing antibody titer is determined through the presence or absence of cytopathic effect in a cell monolayer. The VNT is a relatively inexpensive assay using standard laboratory equipment; however, to perform the assay, cell cultures, significant time, intensive labor, and technical skill are required.
Assuntos
Anticorpos Neutralizantes , Anticorpos Antivirais , Vírus da Doença Hemorrágica Epizoótica , Testes de Neutralização , Testes de Neutralização/métodos , Anticorpos Neutralizantes/imunologia , Anticorpos Neutralizantes/sangue , Animais , Anticorpos Antivirais/imunologia , Anticorpos Antivirais/sangue , Vírus da Doença Hemorrágica Epizoótica/imunologia , Sorogrupo , Infecções por Reoviridae/imunologia , Infecções por Reoviridae/diagnóstico , Infecções por Reoviridae/veterinária , Infecções por Reoviridae/virologiaRESUMO
Risk assessment is the cornerstone of working safely with biological agents. The World Health Organization (WHO) Laboratory Biosafety Manual Fourth Edition Monograph on Risk Assessment provides stepwise guidance for completing a risk assessment, from information gathering and identifying hazards to evaluating the risks, developing, and implementing controls and review.To support the development of a mature safety culture within laboratories, it is important that all staff who handle biological agents understand the fundamentals of risk assessment and receive training in identifying hazards created by their work activities (or tasks) and understand how to mitigate the risks arising from carrying out that work. Any "competent" person may be involved in assessing the risks posed by carrying out an activity. Those closest to the work, who understand the details of the task being undertaken, should be involved in creating the risk assessment. The guidance in this chapter is not just applicable to biosafety professionals, laboratory scientists, or facility managers but can be used by any competent worker familiar with the activity being assessed.This chapter uses the guidance from the WHO to apply the principles of risk assessment to working with Epizootic hemorrhagic disease virus (EHDV), using an example activity-virus isolation from EHDV test samples in cell culture.
Assuntos
Vírus da Doença Hemorrágica Epizoótica , Animais , Vírus da Doença Hemorrágica Epizoótica/isolamento & purificação , Medição de Risco/métodos , Humanos , Gestão de Riscos , Contenção de Riscos Biológicos/métodos , Infecções por Reoviridae/virologia , Infecções por Reoviridae/veterinária , Organização Mundial da Saúde , Orbivirus/genéticaRESUMO
Epizootic hemorrhagic disease virus (EHDV) is an arthropod-borne RNA virus in the genus Orbivirus, family Sedoreoviridae. Globally, seven known EHDV serotypes circulate among ruminant hosts and Culicoides species vectors. A variety of domestic and wild ruminant species are susceptible to EHDV infection, but infection outcome is highly variable between species, as well as between individuals of the same species. Thus, this disease system inherently operates at the wildlife-livestock interface. Domestic cattle are important hosts for EHDV, and while inapparent infection is the most common outcome, reports of clinical disease have increased in some parts of the world. However, fatal infection of cattle is rare. Among wildlife, white-tailed deer (Odocoileus virginianus) are highly susceptible to severe and often fatal disease. Considering the paucity of data and poorly characterized pathology of EHD in cattle, white-tailed deer represent a case study for describing the field signs and necropsy lesions associated with EHD. Here we describe the field signs that commonly define EHD outbreaks in North America, a basic approach to a gross necropsy examination of white-tailed deer, description of the gross lesions that may be present, and diagnostic sample collection. Field investigations of large-scale EHD outbreaks are common in North America. The necropsy examination is an essential tool in the study of disease and when coupled with other disciplines (e.g., virology, immunology, epidemiology) has been fundamentally important to understanding EHD in North America.
Assuntos
Cervos , Vírus da Doença Hemorrágica Epizoótica , Infecções por Reoviridae , Animais , Vírus da Doença Hemorrágica Epizoótica/genética , Infecções por Reoviridae/veterinária , Infecções por Reoviridae/virologia , Cervos/virologia , Autopsia/veterinária , Bovinos , Animais Selvagens/virologiaRESUMO
Real-time RT-PCR for the detection of epizootic hemorrhagic disease virus (EHDV) in clinical samples is a fast and sensitive tool for the diagnosis and confirmation of disease. Several real-time RT-PCR methods have been reported over the last 10 years. In this chapter, we describe seven duplex real-time RT-PCR assays to amplify part of genome segment 2 of EHDV to enable serotype identification. The assay includes the detection of an endogenous control gene-beta-actin.
Assuntos
Vírus da Doença Hemorrágica Epizoótica , Reação em Cadeia da Polimerase em Tempo Real , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Vírus da Doença Hemorrágica Epizoótica/genética , Vírus da Doença Hemorrágica Epizoótica/isolamento & purificação , Vírus da Doença Hemorrágica Epizoótica/classificação , Animais , Reação em Cadeia da Polimerase em Tempo Real/métodos , Reação em Cadeia da Polimerase Via Transcriptase Reversa/métodos , Infecções por Reoviridae/veterinária , Infecções por Reoviridae/virologia , Infecções por Reoviridae/diagnóstico , RNA Viral/genéticaRESUMO
Enzyme-linked immunosorbent assay (ELISA) is a relatively inexpensive, rapid, and high-throughput diagnostic tool to detect antibodies raised against epizootic hemorrhagic disease virus (EHDV) in ruminant serum. While the presence of EHDV antibodies only confirms prior exposure to the virus, it does not conclusively determine infection status. The c-ELISA can be used in conjunction with other diagnostic tests (e.g., real-time PCR) to reinforce diagnosis of infection or as a surveillance tool to support disease control. The EHDV competition ELISA (c-ELISA) described here is a commercial diagnostic assay, recommended by the World Organisation for Animal Health (WOAH), that detects ruminant antibodies against the highly conserved EHDV structural protein, VP7.
Assuntos
Anticorpos Antivirais , Ensaio de Imunoadsorção Enzimática , Vírus da Doença Hemorrágica Epizoótica , Ensaio de Imunoadsorção Enzimática/métodos , Animais , Anticorpos Antivirais/sangue , Anticorpos Antivirais/imunologia , Vírus da Doença Hemorrágica Epizoótica/imunologia , Infecções por Reoviridae/diagnóstico , Infecções por Reoviridae/veterinária , Infecções por Reoviridae/imunologia , Infecções por Reoviridae/virologia , OvinosRESUMO
The titration of viruses onto susceptible cell lines is an important virological technique used to quantify infectious viral titers. It forms an integral component of epizootic hemorrhagic disease virus (EHDV) research, including estimating infectivity, calculating multiplicity of infection, and confirming virus propagation in cell culture. However, the ability to quantify infectious EHDV is also critical for disease control, particularly in the event of an outbreak. Routine EHD diagnostics do not accurately quantify infectious virus, which would allow accurate prediction of the onward transmission risk, but instead are typically more qualitative in nature (e.g., virus isolation) or only quantify viral genome copies (e.g., real-time PCR) which often remain detectable long after infectious virus is cleared from the host.Infectious EHDV titers are typically quantified through the detection of visible cytopathic effect (CPE) in the monolayer of susceptible mammalian cell cultures. However, not all susceptible cell lines demonstrate visible CPE upon EHDV infection, including cell lines such as KC cells, which are derived from the EHDV biological insect vector, Culicoides sonorensis. This chapter presents a comprehensive method for the titration of EHDV-positive samples onto relevant, susceptible mammalian (Vero) and insect (KC) cell lines and describes alternative methods that can be used to visualize EHDV infection, by CPE or immunofluorescent labeling of viral proteins, to enable the calculation of infectious EHDV titers.
Assuntos
Vírus da Doença Hemorrágica Epizoótica , Vírus da Doença Hemorrágica Epizoótica/isolamento & purificação , Vírus da Doença Hemorrágica Epizoótica/genética , Animais , Linhagem Celular , Carga Viral , Infecções por Reoviridae/virologia , Infecções por Reoviridae/veterinária , Efeito Citopatogênico Viral , Cultura de Vírus/métodosRESUMO
The emergence of EHDV in Europe during the autumn of 2022 reinforces the need for molecular tools (RT-PCR) for rapid detection of animals infected with this virus. Viral genome testing can be performed on whole blood under anticoagulant, spleen, and bloody organ homogenates from ruminants. It can also be performed on cell culture following viral isolation tests. Various so-called classical or end-point RT-PCRs will be described, which permit the amplification of a part of the viral genome (targeting segment 7) allowing the detection of EHDV whatever the serotype (pan-RT-PCR) and also to amplify a portion of the gene coding the viral protein (VP) 2 enabling serotyping. The PCR amplification products are visualized by agarose gel electrophoresis. Sequencing of the type-specific RT-PCR amplification products allows for the serotype of the virus to be determined.
Assuntos
Vírus da Doença Hemorrágica Epizoótica , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Animais , Vírus da Doença Hemorrágica Epizoótica/genética , Vírus da Doença Hemorrágica Epizoótica/isolamento & purificação , Vírus da Doença Hemorrágica Epizoótica/classificação , Reação em Cadeia da Polimerase Via Transcriptase Reversa/métodos , Infecções por Reoviridae/veterinária , Infecções por Reoviridae/virologia , Infecções por Reoviridae/diagnóstico , RNA Viral/genética , Genoma Viral , Sorotipagem/métodosRESUMO
Real-time reverse transcription-polymerase chain reaction (real-time RT-PCR) has become an essential tool in rapid and reliable detection of animal diseases such as epizootic hemorrhagic disease (EHD). Here we provide a protocol for the detection of epizootic hemorrhagic disease virus (EHDV) genetic material in blood and tissue samples, using a real-time RT-PCR that targets a conserved region in segment 9 of the EHDV genome. This protocol can be used to detect up to approximately 90 samples in a single run and can be completed in less than 4 h.
Assuntos
Vírus da Doença Hemorrágica Epizoótica , Reação em Cadeia da Polimerase em Tempo Real , Infecções por Reoviridae , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Vírus da Doença Hemorrágica Epizoótica/genética , Vírus da Doença Hemorrágica Epizoótica/isolamento & purificação , Animais , Reação em Cadeia da Polimerase em Tempo Real/métodos , Infecções por Reoviridae/diagnóstico , Infecções por Reoviridae/veterinária , Infecções por Reoviridae/virologia , Reação em Cadeia da Polimerase Via Transcriptase Reversa/métodos , RNA Viral/genéticaRESUMO
Reverse transcriptase loop-mediated isothermal amplification (RT-LAMP) is a molecular diagnostic assay that is particularly useful for the detection of viral diseases of livestock. A major advantage of RT-LAMP is that it can be used either as a rapid field test or as a high-throughput screening tool in veterinary laboratories, with sensitivity comparable to the real-time RT-PCR assay. Unlike conventional or qPCR, RT-LAMP uses a strand displacement polymerase and a set of four to six primers that bind to several regions of the target nucleic acid. Amplification occurs without thermal cycling, and coupled with the numerous primers, RT-LAMP offers a rapid and highly specific molecular assay. In this chapter, we describe the RT-LAMP protocol for the detection of epizootic hemorrhagic disease virus (EHDV) as a low-cost, specific, and sensitive screening tool in veterinary diagnostic laboratories. We also provide guidance on how to adapt the RT-LAMP assay for rapid field testing.
Assuntos
Vírus da Doença Hemorrágica Epizoótica , Técnicas de Diagnóstico Molecular , Técnicas de Amplificação de Ácido Nucleico , Infecções por Reoviridae , Técnicas de Amplificação de Ácido Nucleico/métodos , Animais , Vírus da Doença Hemorrágica Epizoótica/genética , Vírus da Doença Hemorrágica Epizoótica/isolamento & purificação , Técnicas de Diagnóstico Molecular/métodos , Infecções por Reoviridae/diagnóstico , Infecções por Reoviridae/veterinária , Infecções por Reoviridae/virologia , Sensibilidade e Especificidade , Primers do DNA/genética , RNA Viral/genéticaRESUMO
Virus isolation is used to assist in the diagnosis and confirmation of viral infections. Successful isolation of a virus is highly dependent upon the quality of starting material. Here we describe the preparation and isolation of epizootic hemorrhagic disease virus (EHDV) from blood and tissue samples in tissue culture flasks (TCFs) through the inoculation of susceptible cell lines including Vero, BHK, and KC cells.
Assuntos
Vírus da Doença Hemorrágica Epizoótica , Animais , Vírus da Doença Hemorrágica Epizoótica/isolamento & purificação , Chlorocebus aethiops , Linhagem Celular , Células Vero , Infecções por Reoviridae/virologia , Infecções por Reoviridae/veterinária , Técnicas de Cultura de Células/métodos , Cricetinae , Cultura de Vírus/métodosRESUMO
Next-generation sequencing (NGS) technologies are continuously being developed and are becoming a more cost-effective tool for the characterization of viral genomes. Whole genome sequencing of segmented viruses, such as epizootic hemorrhagic disease virus (EHDV), provides insights into the molecular epidemiology as well as such viral evolutionary mechanisms as genetic reassortment. Here, we present a detailed method for obtaining full genome sequence data for EHDV using Illumina technology. The protocol includes details from RNA extraction and purification, the synthesis of cDNA, sequencing library preparation, to genome assembly.
Assuntos
Genoma Viral , Vírus da Doença Hemorrágica Epizoótica , Sequenciamento de Nucleotídeos em Larga Escala , Sequenciamento Completo do Genoma , Vírus da Doença Hemorrágica Epizoótica/genética , Vírus da Doença Hemorrágica Epizoótica/isolamento & purificação , Sequenciamento de Nucleotídeos em Larga Escala/métodos , Sequenciamento Completo do Genoma/métodos , Animais , RNA Viral/genética , Biblioteca Gênica , Infecções por Reoviridae/virologia , Infecções por Reoviridae/veterináriaRESUMO
Molecular methods are routinely used for the differential diagnosis and genetic characterization of viral disease of livestock. Real-time, quantitative PCR (qPCR) allows RNA/DNA sequence detection and quantification and is considered the gold standard diagnostic method for most viruses. However, Sanger sequencing offers additional information and opportunity to differentiate closely related virus strains and/or serotypes, by providing the full sequence of a genetic region of interest. Therefore, to determine epizootic hemorrhagic disease virus (EHDV) serotype or identify additional genetic markers, end-point RT-PCR can be performed on EHDV-positive clinical samples, followed by Sanger sequencing and data analysis. Here we describe a detailed method for the molecular characterization of EHDV serotype using Sanger sequencing.
Assuntos
Vírus da Doença Hemorrágica Epizoótica , Infecções por Reoviridae , Sorotipagem , Vírus da Doença Hemorrágica Epizoótica/genética , Vírus da Doença Hemorrágica Epizoótica/classificação , Animais , Sorotipagem/métodos , Infecções por Reoviridae/virologia , Infecções por Reoviridae/veterinária , RNA Viral/genética , Sorogrupo , Análise de Sequência de DNA/métodos , Reação em Cadeia da Polimerase em Tempo Real/métodosRESUMO
Epizootic hemorrhagic disease virus (EHDV) is transmitted by Culicoides biting midges. Studies aiming to predict the likely spread of EHDV require an understanding of the viral infection and replication kinetics within these insects, including the proportion of the insect population that are able to support virus transmission. Here, we describe methods for the infection of Culicoides with EHDV in the laboratory via oral infection using an artificial membrane system or a cotton pledget and intrathoracic (IT) inoculation. Each method can be used to explore determinants of vector competence of Culicoides species and populations for EHDV.
Assuntos
Ceratopogonidae , Vírus da Doença Hemorrágica Epizoótica , Insetos Vetores , Infecções por Reoviridae , Animais , Ceratopogonidae/virologia , Vírus da Doença Hemorrágica Epizoótica/fisiologia , Insetos Vetores/virologia , Infecções por Reoviridae/transmissão , Infecções por Reoviridae/virologia , Infecções por Reoviridae/veterináriaRESUMO
Mammalian orthoreoviruses (MRVs), belonging to the genus Orthoreovirus in the family Spinareoviridae, possess a double-stranded RNA segmented genome. Due to the segmented nature of their genome, MRVs are prone to gene reassortment, which allows for evolutionary diversification. Recently, a genotyping system for each MRV gene segment was proposed based on nucleotide differences. In the present study, MRVs were isolated from the fecal samples of Japanese Black cattle kept on a farm in Japan. Complete genome sequencing and analysis of 41 MRV isolates revealed that these MRVs shared almost identical sequences in the L1, L2, L3, S3, and S4 gene segments, while two different sequences were found in the S1, M1, M2, M3, and S2 gene segments. By plaque cloning, at least six genetic constellation patterns were identified, indicating the occurrence of multiple inter- (S1 and M2) and intra- (M1, M3, and S2) reassortment events. This paper represents the first report describing multiple reassortant MRVs on a single cattle farm. These MRV gene segments exhibited sequence similarity to those of MRVs isolated from cattle in the U.S. and China, rather than to MRVs previously isolated in Japan. Genotypes consisting solely of bovine MRVs were observed in the L1, M1, and M2 segments, suggesting that they might have evolved within the cattle population.
Assuntos
Fazendas , Genoma Viral , Genótipo , Orthoreovirus de Mamíferos , Filogenia , Vírus Reordenados , Animais , Bovinos , Vírus Reordenados/genética , Vírus Reordenados/isolamento & purificação , Japão , Orthoreovirus de Mamíferos/genética , Orthoreovirus de Mamíferos/isolamento & purificação , Orthoreovirus de Mamíferos/classificação , Doenças dos Bovinos/virologia , Infecções por Reoviridae/veterinária , Infecções por Reoviridae/virologia , Fezes/virologiaRESUMO
BF/C2 is a crucial molecule in the coagulation complement cascade pathway and plays a significant role in the immune response of grass carp through the classical, alternative, and lectin pathways during GCRV infection. In vivo experiments demonstrated that the mRNA expression levels of BF/C2 (A, B) in grass carp positively correlated with GCRV viral replication at various stages of infection. Excessive inflammation leading to death coincided with peak levels of BF/C2 (A, B) mRNA expression and GCRV viral replication. Correspondingly, BF/C2 (A, B) recombinant protein, CIK cells and GCRV co-incubation experiments yielded similar findings. Therefore, 3 h (incubation period) and 9 h (death period) were selected as critical points for this study. Transcriptome sequencing analysis revealed significant differences in the expression of BF/C2A and BF/C2B during different stages of CIK infection with GCRV and compared to the blank control group (PBS). Specifically, the BF/C2A_3 and BF/C2A_9 groups exhibited 2729 and 2228 differentially expressed genes (DEGs), respectively, with 1436 upregulated and 1293 downregulated in the former, and 1324 upregulated and 904 downregulated in the latter. The BF/C2B_3 and BF/C2B_9 groups showed 2303 and 1547 DEGs, respectively, with 1368 upregulated and 935 downregulated in the former, and 818 upregulated and 729 downregulated in the latter. KEGG functional enrichment analysis of these DEGs identified shared pathways between BF/C2A and PBS groups at 3 and 9 h, including the C-type lectin receptor signaling pathway, protein processing in the endoplasmic reticulum, Toll-like receptor signaling pathway, Salmonella infection, apoptosis, tight junction, and adipocytokine signaling pathway. Additionally, the BF/C2B groups at 3 and 9 h shared pathways related to protein processing in the endoplasmic reticulum, glycolysis/gluconeogenesis, and biosynthesis of amino acids. The mRNA levels of these DEGs were validated in cellular models, confirming consistency with the sequencing results. In addition, the mRNA expression levels of these candidate genes (mapk1, il1b, rela, nfkbiab, akt3a, hyou1, hsp90b1, dnajc3a et al.) in the head kidney, kidney, liver and spleen of grass carp immune tissue were significantly different from those of the control group by BF/C2 (A, B) protein injection in vivo. These candidate genes play an important role in the response of BF/C2 (A, B) to GCRV infection and it also further confirmed that BF/C2 (A, B) of grass carp plays an important role in coping with GCRV infection.
Assuntos
Carpas , Doenças dos Peixes , Proteínas de Peixes , Infecções por Reoviridae , Reoviridae , Animais , Carpas/genética , Carpas/virologia , Carpas/imunologia , Doenças dos Peixes/virologia , Doenças dos Peixes/imunologia , Doenças dos Peixes/genética , Infecções por Reoviridae/veterinária , Infecções por Reoviridae/imunologia , Infecções por Reoviridae/genética , Infecções por Reoviridae/virologia , Proteínas de Peixes/genética , Proteínas de Peixes/metabolismo , Reoviridae/fisiologia , Perfilação da Expressão Gênica , Transcriptoma , Replicação Viral , Regulação da Expressão GênicaRESUMO
Similar to other RNA viruses, grass carp reovirus, the causative agent of the hemorrhagic disease, replicates in cytoplasmic viral inclusion bodies (VIBs), orchestrated by host proteins and lipids. The host pathways that facilitate the formation and function of GCRV VIBs are poorly understood. This work demonstrates that GCRV manipulates grass carp oxysterol binding protein 1 (named as gcOSBP1) and vesicle-associated membrane protein-associated protein A/B (named as gcVAP-A/B), 3 components of cholesterol transport pathway, to generate VIBs. By siRNA-mediated knockdown, we demonstrate that gcOSBP1 is an essential host factor for GCRV replication. We reveal that the nonstructural proteins NS80 and NS38 of GCRV interact with gcOSBP1, and that the gcOSBP1 is recruited by NS38 and NS80 for promoting the generation of VIBs. gcOSBP1 increases the expression of gcVAP-A/B and promotes the accumulation of intracellular cholesterol. gcOSBP1 also interacts with gcVAP-A/B for forming gcOSBP1-gcVAP-A/B complexes, which contribute to enhance the accumulation of intracellular cholesterol and gcOSBP1-mediated generation of VIBs. Inhibiting cholesterol accumulation by lovastatin can completely abolish the effects of gcOSBP1 and/or gcVAP-A/B in promoting GCRV infection, suggesting that cholesterol accumulation is vital for gcOSBP1- and/or gcVAP-A/B-mediated GCRV replication. Thus, our results, which highlight that gcOSBP1 functions in the replication of GCRV via its interaction with essential viral proteins for forming VIBs and with host gcVAP-A/B, provide key molecular targets for obtaining anti-hemorrhagic disease grass carp via gene editing technology.
Assuntos
Carpas , Colesterol , Corpos de Inclusão Viral , Receptores de Esteroides , Reoviridae , Replicação Viral , Animais , Reoviridae/fisiologia , Carpas/virologia , Carpas/metabolismo , Corpos de Inclusão Viral/metabolismo , Colesterol/metabolismo , Receptores de Esteroides/metabolismo , Doenças dos Peixes/virologia , Doenças dos Peixes/metabolismo , Doenças dos Peixes/imunologia , Interações Hospedeiro-Patógeno , Infecções por Reoviridae/veterinária , Infecções por Reoviridae/metabolismo , Infecções por Reoviridae/virologia , Proteínas de Peixes/metabolismo , Proteínas de Peixes/genética , Proteínas não Estruturais Virais/metabolismo , Proteínas não Estruturais Virais/genéticaRESUMO
In this study, we provide a genomic description of the first isolation of the Umattila virus (UMAV) in Brazil. The virus was obtained from the blood of a bird (Turdus fumigatus) and isolated in a C6/36 cell culture. The viral genome contains ten segments, and its organization is characteristic of viruses of the genus Orbivirus (family Sedoreoviridae). The coding region of each segment was sequenced, demonstrating the nucleotide identity with UMAV. The phylogenetic inference results were in line with these findings and demonstrated the formation of two distinct monophyletic clades containing strains isolated around the world, where our isolate, belonging to the same clade as the prototype strain, was allocated to a different subclade, highlighting the genetic divergence between them. This work reports the first isolation of UMAV in Brazil, and due to the scarcity of information on this viral agent in the scientific literature, it is essential to carry out further studies to better understand its epidemiology, dispersion, and, in particular, its interactions with vertebrate hosts, vectors, and the environment.
Assuntos
Genoma Viral , Orbivirus , Filogenia , Brasil , Animais , Orbivirus/isolamento & purificação , Orbivirus/genética , Orbivirus/classificação , Infecções por Reoviridae/virologia , Infecções por Reoviridae/veterinária , Aves/virologia , Doenças das Aves/virologia , RNA Viral/genética , Linhagem CelularRESUMO
Avian orthoreviruses have become a global challenge to the poultry industry, causing significant economic impacts on commercial poultry. Avian reoviruses (ARVs) are resistant to heat, proteolytic enzymes, a wide range of pH values, and disinfectants, so keeping chicken farms free of ARV infections is difficult. This review focuses on the global prevalence of ARVs and associated clinical signs and symptoms. The most common signs and symptoms include tenosynovitis/arthritis, malabsorption syndrome, runting-stunting syndrome, and respiratory diseases. Moreover, this review also focused on the characterization of ARVs in genotypic clusters (I-VI) and their relation to tissue tropism or viral distribution. The prevailing strains of ARV in Africa belong to all genotypic clusters (GCs) except for GC VI, whereas all GCs are present in Asia and the Americas. In addition, all ARV strains are associated with or belong to GC I-VI in Europe. Moreover, in Oceania, only GC V and VI are prevalent. This review also showed that, regardless of the genotypic cluster, tenosynovitis/arthritis was the predominant clinical manifestation, indicating its universal occurrence across all clusters. Globally, most avian reovirus infections can be prevented by vaccination against four major strains: S1133, 1733, 2408, and 2177. Nevertheless, these vaccines may not a provide sufficient defense against field isolates. Due to the increase in the number of ARV variants, classical vaccine approaches are being developed depending on the degree of antigenic similarity between the vaccine and field strains, which determines how successful the vaccination will be. Moreover, there is a need to look more closely at the antigenic and pathogenic properties of reported ARV strains. The information acquired will aid in the selection of more effective vaccine strains in combination with biosecurity and farm management methods to prevent ARV infections.
Assuntos
Genótipo , Orthoreovirus Aviário , Doenças das Aves Domésticas , Infecções por Reoviridae , Orthoreovirus Aviário/genética , Orthoreovirus Aviário/classificação , Animais , Doenças das Aves Domésticas/virologia , Doenças das Aves Domésticas/epidemiologia , Infecções por Reoviridae/veterinária , Infecções por Reoviridae/epidemiologia , Infecções por Reoviridae/virologia , Filogenia , Galinhas/virologia , Prevalência , Aves Domésticas/virologiaRESUMO
In this issue of Cell Host & Microbe, Shang et al. identify murine neuropilin 1 as a host factor that binds reovirus particles, directing cell entry and contributing to viral dissemination and neurovirulence. This study highlights the reovirus model system to investigate host receptors and their significance in viral pathogenesis.